-DOCSTART-	O

IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
through	NN	O	O
CD28	NN	O	B-protein
requires	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
production	NN	O	O
by	NN	O	O
5-lipoxygenase	NN	O	B-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
surface	NN	O	I-protein
receptor	NN	O	I-protein
provides	NN	O	O
a	NN	O	O
major	NN	O	O
costimulatory	NN	O	O
signal	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
resulting	NN	O	O
in	NN	O	O
enhanced	NN	O	O
production	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
we	NN	O	O
show	NN	O	O
that	NN	O	O
CD28	NN	O	B-protein
ligation	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
rapid	NN	O	O
intracellular	NN	O	O
formation	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
(	NN	O	O
ROIs	NN	O	O
)	NN	O	O
which	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
CD28	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
/	NN	O	O
CD28-responsive	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Delineation	NN	O	O
of	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
signaling	NN	O	O
cascade	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
involve	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
phospholipase	NN	O	B-protein
A2	NN	O	I-protein
and	NN	O	O
5-lipoxygenase	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
lipoxygenase	NN	O	B-protein
metabolites	NN	O	I-protein
activate	NN	O	O
ROI	NN	O	O
formation	NN	O	O
which	NN	O	O
then	NN	O	O
induce	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
via	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
should	NN	O	O
be	NN	O	O
useful	NN	O	O
for	NN	O	O
therapeutic	NN	O	O
strategies	NN	O	O
and	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
immunosuppressants	NN	O	O
targeting	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
costimulatory	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
peri-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
mediates	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
2	NN	O	I-DNA
enhancer	NN	O	I-DNA
activation	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
2	NN	O	O
(	NN	O	O
HIV-2	NN	O	O
)	NN	O	O
,	NN	O	O
like	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
causes	NN	O	O
AIDS	NN	O	O
and	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
AIDS	NN	O	O
cases	NN	O	O
primarily	NN	O	O
in	NN	O	O
West	NN	O	O
Africa	NN	O	O
.	NN	O	O

HIV-1	NN	O	O
and	NN	O	O
HIV-2	NN	O	O
display	NN	O	O
significant	NN	O	O
differences	NN	O	O
in	NN	O	O
nucleic	NN	O	O
acid	NN	O	O
sequence	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
natural	NN	O	O
history	NN	O	O
of	NN	O	O
clinical	NN	O	O
disease	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
these	NN	O	O
differences	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
enhancer/promoter	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
HIV-2	NN	O	O
functions	NN	O	O
quite	NN	O	O
differently	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

Whereas	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
following	NN	O	O
T-cell	NN	O	O
stimulation	NN	O	O
is	NN	O	O
mediated	NN	O	O
largely	NN	O	O
through	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
to	NN	O	O
two	NN	O	O
adjacent	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
,	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
four	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
:	NN	O	O
a	NN	O	O
single	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
two	NN	O	O
purine-rich	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
PuB1	NN	O	B-DNA
and	NN	O	O
PuB2	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
pets	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	O
cis-acting	NN	O	B-DNA
element	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
HIV-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
immediately	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
designated	NN	O	O
peri-kappa	NN	O	B-DNA
B	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
site	NN	O	O
is	NN	O	O
conserved	NN	O	O
among	NN	O	O
isolates	NN	O	O
of	NN	O	O
HIV-2	NN	O	O
and	NN	O	O
the	NN	O	O
closely	NN	O	O
related	NN	O	O
simian	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
,	NN	O	O
and	NN	O	O
transfection	NN	O	O
assays	NN	O	O
show	NN	O	O
this	NN	O	O
site	NN	O	O
to	NN	O	O
mediate	NN	O	O
HIV-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
activation	NN	O	O
following	NN	O	O
stimulation	NN	O	O
of	NN	O	O
monocytic	NN	O	B-cell_line
but	NN	O	O
not	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
description	NN	O	O
of	NN	O	O
an	NN	O	O
HIV-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
which	NN	O	O
displays	NN	O	O
such	NN	O	O
monocyte	NN	O	O
specificity	NN	O	O
,	NN	O	O
and	NN	O	O
no	NN	O	O
comparable	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
clearly	NN	O	O
defined	NN	O	O
for	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

While	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
from	NN	O	O
both	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
binds	NN	O	O
the	NN	O	O
peri-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
suggest	NN	O	O
that	NN	O	O
either	NN	O	O
a	NN	O	O
different	NN	O	O
protein	NN	O	O
binds	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
versus	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
that	NN	O	O
the	NN	O	O
protein	NN	O	O
recognizing	NN	O	O
this	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
undergoes	NN	O	O
differential	NN	O	O
modification	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
thus	NN	O	O
supporting	NN	O	O
the	NN	O	O
transfection	NN	O	O
data	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
while	NN	O	O
specific	NN	O	O
constitutive	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
peri-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
is	NN	O	O
seen	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
induces	NN	O	O
additional	NN	O	O
,	NN	O	O
specific	NN	O	O
binding	NN	O	O
.	NN	O	O

Understanding	NN	O	O
the	NN	O	O
monocyte-specific	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
peri-kappa	NN	O	B-protein
B	NN	O	I-protein
factor	NN	O	I-protein
may	NN	O	O
ultimately	NN	O	O
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
different	NN	O	O
role	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
play	NN	O	O
in	NN	O	O
HIV	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

E1A	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
induces	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
killing	NN	O	O
by	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
following	NN	O	O
immortalization	NN	O	O
but	NN	O	O
not	NN	O	O
adenovirus	NN	O	O
infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Adenovirus	NN	O	O
(	NN	O	O
Ad	NN	O	O
)	NN	O	O
infection	NN	O	O
and	NN	O	O
E1A	NN	O	B-protein
transfection	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
model	NN	O	O
changes	NN	O	O
in	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
NK	NN	O	O
cell	NN	O	O
killing	NN	O	O
caused	NN	O	O
by	NN	O	O
transient	NN	O	O
vs	NN	O	O
stable	NN	O	O
E1A	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Only	NN	O	O
stably	NN	O	O
transfected	NN	O	B-cell_line
target	NN	O	I-cell_line
cells	NN	O	I-cell_line
exhibited	NN	O	O
cytolytic	NN	O	O
susceptibility	NN	O	O
,	NN	O	O
despite	NN	O	O
expression	NN	O	O
of	NN	O	O
equivalent	NN	O	O
levels	NN	O	O
of	NN	O	O
E1A	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
Ad-infected	NN	O	B-cell_line
targets	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
inability	NN	O	O
of	NN	O	O
E1A	NN	O	B-DNA
gene	NN	O	I-DNA
products	NN	O	O
to	NN	O	O
induce	NN	O	O
cytolytic	NN	O	O
susceptibility	NN	O	O
during	NN	O	O
infection	NN	O	O
was	NN	O	O
not	NN	O	O
explained	NN	O	O
by	NN	O	O
an	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
viral	NN	O	O
infection	NN	O	O
on	NN	O	O
otherwise	NN	O	O
susceptible	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
by	NN	O	O
viral	NN	O	B-DNA
gene	NN	O	I-DNA
effects	NN	O	O
on	NN	O	O
class	NN	O	B-protein
I	NN	O	I-protein
MHC	NN	O	I-protein
antigen	NN	O	I-protein
expression	NN	O	O
on	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
differential	NN	O	O
effect	NN	O	O
of	NN	O	O
E1A	NN	O	B-protein
expression	NN	O	O
on	NN	O	O
the	NN	O	O
cytolytic	NN	O	O
phenotypes	NN	O	O
of	NN	O	O
infected	NN	O	O
and	NN	O	O
stably	NN	O	O
transfected	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
suggests	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
provide	NN	O	O
an	NN	O	O
effective	NN	O	O
immunologic	NN	O	O
barrier	NN	O	O
against	NN	O	O
the	NN	O	O
in	NN	O	O
vivo	NN	O	O
survival	NN	O	O
and	NN	O	O
neoplastic	NN	O	O
progression	NN	O	O
of	NN	O	O
E1A-immortalized	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
may	NN	O	O
emerge	NN	O	O
from	NN	O	O
the	NN	O	O
reservoir	NN	O	O
of	NN	O	O
persistently	NN	O	B-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
human	NN	O	O
host	NN	O	O
.	NN	O	O

-DOCSTART-	O

Distinct	NN	O	O
signaling	NN	O	O
properties	NN	O	O
identify	NN	O	O
functionally	NN	O	O
different	NN	O	O
CD4	NN	O	B-protein
epitopes	NN	O	I-protein
.	NN	O	O

The	NN	O	O
CD4	NN	O	B-protein
coreceptor	NN	O	I-protein
interacts	NN	O	O
with	NN	O	O
non-polymorphic	NN	O	B-protein
regions	NN	O	I-protein
of	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
on	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
contributes	NN	O	O
to	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
triggering	NN	O	O
on	NN	O	O
T	NN	O	O
cell	NN	O	O
activating	NN	O	O
signals	NN	O	O
in	NN	O	O
a	NN	O	O
lymphoma	NN	O	O
model	NN	O	O
using	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
(	NN	O	O
mAb	NN	O	B-protein
)	NN	O	O
which	NN	O	O
recognize	NN	O	O
different	NN	O	O
CD4	NN	O	B-protein
epitopes	NN	O	I-protein
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
CD4	NN	O	B-protein
triggering	NN	O	O
delivers	NN	O	O
signals	NN	O	O
capable	NN	O	O
of	NN	O	O
activating	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
which	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
interleukin-2	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Whereas	NN	O	O
different	NN	O	O
anti-CD4	NN	O	B-protein
mAb	NN	O	I-protein
or	NN	O	O
HIV-1	NN	O	B-protein
gp120	NN	O	I-protein
could	NN	O	O
all	NN	O	O
trigger	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
p56lck	NN	O	B-protein
and	NN	O	O
p59fyn	NN	O	B-protein
and	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
Shc	NN	O	B-protein
adaptor	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
mediates	NN	O	O
signals	NN	O	O
to	NN	O	O
Ras	NN	O	B-protein
,	NN	O	O
they	NN	O	O
differed	NN	O	O
significantly	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

Lack	NN	O	O
of	NN	O	O
full	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
could	NN	O	O
be	NN	O	O
correlated	NN	O	O
to	NN	O	O
a	NN	O	O
dramatically	NN	O	O
reduced	NN	O	O
capacity	NN	O	O
to	NN	O	O
induce	NN	O	O
calcium	NN	O	O
flux	NN	O	O
and	NN	O	O
could	NN	O	O
be	NN	O	O
complemented	NN	O	O
with	NN	O	O
a	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
identify	NN	O	O
functionally	NN	O	O
distinct	NN	O	O
epitopes	NN	O	B-protein
on	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
coreceptor	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
Ras/protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
and	NN	O	O
calcium	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

Ligand-dependent	NN	O	O
repression	NN	O	O
of	NN	O	O
the	NN	O	O
erythroid	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
by	NN	O	O
the	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

High-dose	NN	O	O
estrogen	NN	O	O
administration	NN	O	O
induces	NN	O	O
anemia	NN	O	O
in	NN	O	O
mammals	NN	O	O
.	NN	O	O

In	NN	O	O
chickens	NN	O	O
,	NN	O	O
estrogens	NN	O	O
stimulate	NN	O	O
outgrowth	NN	O	O
of	NN	O	O
bone	NN	O	B-cell_type
marrow-derived	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
delay	NN	O	O
their	NN	O	O
maturation	NN	O	O
.	NN	O	O

This	NN	O	O
delay	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
many	NN	O	O
erythroid	NN	O	B-DNA
cell-specific	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
alpha-	NN	O	B-DNA
and	NN	O	I-DNA
beta-globin	NN	O	I-DNA
,	NN	O	O
band	NN	O	O
3	NN	O	O
,	NN	O	O
band	NN	O	O
4.1	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
erythroid	NN	O	B-protein
cell-specific	NN	O	I-protein
histone	NN	O	I-protein
H5	NN	O	B-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
estrogens	NN	O	O
also	NN	O	O
reduce	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
primary	NN	O	B-cell_line
human	NN	O	I-cell_line
bone	NN	O	I-cell_line
marrow	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
address	NN	O	O
potential	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
estrogens	NN	O	O
suppress	NN	O	O
erythropoiesis	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
their	NN	O	O
effects	NN	O	O
on	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
an	NN	O	O
erythroid	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
participates	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
erythroid	NN	O	B-DNA
cell-specific	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
full	NN	O	O
maturation	NN	O	O
of	NN	O	O
erythrocytes	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
is	NN	O	O
strongly	NN	O	O
repressed	NN	O	O
by	NN	O	O
the	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
in	NN	O	O
a	NN	O	O
ligand-dependent	NN	O	O
manner	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
repression	NN	O	O
is	NN	O	O
reversible	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
4-hydroxytamoxifen	NN	O	O
.	NN	O	O

ER	NN	O	B-protein
-mediated	NN	O	O
repression	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
activity	NN	O	O
occurs	NN	O	O
on	NN	O	O
an	NN	O	O
artificial	NN	O	B-DNA
promoter	NN	O	I-DNA
containing	NN	O	O
a	NN	O	O
single	NN	O	O
GATA-binding	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
an	NN	O	O
intact	NN	O	B-DNA
promoter	NN	O	I-DNA
which	NN	O	O
is	NN	O	O
normally	NN	O	O
regulated	NN	O	O
by	NN	O	O
GATA-1	NN	O	B-protein
.	NN	O	O

GATA-1	NN	O	B-protein
and	NN	O	O
ER	NN	O	B-protein
bind	NN	O	O
to	NN	O	O
each	NN	O	O
other	NN	O	O
in	NN	O	O
vitro	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
DNA	NN	O	O
.	NN	O	O

In	NN	O	O
coimmunoprecipitation	NN	O	O
experiments	NN	O	O
using	NN	O	O
transfected	NN	O	O
COS	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
ER	NN	O	B-protein
associate	NN	O	O
in	NN	O	O
a	NN	O	O
ligand-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Mapping	NN	O	O
experiments	NN	O	O
indicate	NN	O	O
that	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
ER	NN	O	B-protein
form	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
contacts	NN	O	O
,	NN	O	O
which	NN	O	O
involve	NN	O	O
the	NN	O	O
finger	NN	O	O
region	NN	O	O
and	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
GATA-1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
speculate	NN	O	O
that	NN	O	O
estrogens	NN	O	O
exert	NN	O	O
effects	NN	O	O
on	NN	O	O
erythropoiesis	NN	O	O
by	NN	O	O
modulating	NN	O	O
GATA-1	NN	O	B-protein
activity	NN	O	O
through	NN	O	O
protein-protein	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
ER	NN	O	B-protein
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Mouse	NN	O	B-DNA
interleukin-2	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Interleukin-1	NN	O	B-protein
and	NN	O	O
interleukin-2	NN	O	B-protein
control	NN	O	O
transcription	NN	O	O
via	NN	O	O
distinct	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
control	NN	O	O
IL-2	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
CD4-CD8-	NN	O	B-cell_line
murine	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
precursors	NN	O	I-cell_line
.	NN	O	O

Here	NN	O	O
we	NN	O	O
map	NN	O	O
the	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
that	NN	O	O
mediate	NN	O	O
interleukin	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
using	NN	O	O
a	NN	O	O
thymic	NN	O	B-cell_line
lymphoma-derived	NN	O	I-cell_line
hybridoma	NN	O	I-cell_line
(	NN	O	O
PC60	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
to	NN	O	O
stimulation	NN	O	O
by	NN	O	O
IL-1	NN	O	B-protein
+	NN	O	O
IL-2	NN	O	B-protein
is	NN	O	O
biphasic	NN	O	O
.	NN	O	O

IL-1	NN	O	B-protein
induces	NN	O	O
a	NN	O	O
rapid	NN	O	O
,	NN	O	O
protein	NN	O	O
synthesis-independent	NN	O	O
appearance	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-RNA
alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
that	NN	O	O
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
primes	NN	O	O
cells	NN	O	O
to	NN	O	O
become	NN	O	O
IL-2	NN	O	B-protein
responsive	NN	O	O
and	NN	O	O
thereby	NN	O	O
prepares	NN	O	O
the	NN	O	O
second	NN	O	O
phase	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
IL-2	NN	O	B-protein
induces	NN	O	O
a	NN	O	O
100-fold	NN	O	O
further	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-2R	NN	O	B-RNA
alpha	NN	O	I-RNA
transcripts	NN	O	I-RNA
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
show	NN	O	O
that	NN	O	O
several	NN	O	O
elements	NN	O	O
in	NN	O	O
the	NN	O	O
promoter-proximal	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
contribute	NN	O	O
to	NN	O	O
IL-1	NN	O	B-protein
responsiveness	NN	O	O
,	NN	O	O
most	NN	O	O
importantly	NN	O	O
an	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
conserved	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
mouse	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

IL-2	NN	O	B-protein
responsiveness	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
depends	NN	O	O
on	NN	O	O
a	NN	O	O
78-nucleotide	NN	O	B-DNA
segment	NN	O	I-DNA
1.3	NN	O	O
kilobases	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
transcription	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
segment	NN	O	O
functions	NN	O	O
as	NN	O	O
an	NN	O	O
IL-2-inducible	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
lies	NN	O	O
within	NN	O	O
a	NN	O	O
region	NN	O	O
that	NN	O	O
becomes	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
hypersensitive	NN	O	O
in	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
which	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
has	NN	O	O
been	NN	O	O
induced	NN	O	O
.	NN	O	O

IL-2	NN	O	B-protein
responsiveness	NN	O	O
requires	NN	O	O
three	NN	O	O
distinct	NN	O	O
elements	NN	O	B-DNA
within	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
.	NN	O	O

Two	NN	O	O
of	NN	O	O
these	NN	O	O
are	NN	O	O
potential	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Hematopoietic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
commitment	NN	O	O
:	NN	O	O
role	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

This	NN	O	O
review	NN	O	O
focuses	NN	O	O
on	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
hematopoietic	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
.	NN	O	O

A	NN	O	O
brief	NN	O	O
introduction	NN	O	O
to	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
and	NN	O	O
asymmetric	NN	O	O
cell	NN	O	O
division	NN	O	O
is	NN	O	O
followed	NN	O	O
by	NN	O	O
a	NN	O	O
discussion	NN	O	O
of	NN	O	O
several	NN	O	O
methods	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
important	NN	O	O
in	NN	O	O
specifying	NN	O	O
hematopoietic	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
.	NN	O	O

Next	NN	O	O
is	NN	O	O
presented	NN	O	O
a	NN	O	O
discussion	NN	O	O
of	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
embryonic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
analysis	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
and	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
targeted	NN	O	O
gene	NN	O	O
disruption	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
status	NN	O	O
of	NN	O	O
our	NN	O	O
current	NN	O	O
knowledge	NN	O	O
concerning	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
commitment	NN	O	O
to	NN	O	O
erythroid	NN	O	B-cell_type
,	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
and	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
is	NN	O	O
summarized	NN	O	O
.	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	O
virus	NN	O	O
replicative	NN	O	O
gene	NN	O	O
transcription	NN	O	O
during	NN	O	O
de	NN	O	O
novo	NN	O	O
infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
:	NN	O	O
simultaneous	NN	O	O
early	NN	O	O
expression	NN	O	O
of	NN	O	O
BZLF-1	NN	O	B-protein
and	NN	O	O
its	NN	O	O
repressor	NN	O	O
RAZ	NN	O	B-protein
.	NN	O	O

Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
infect	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
and	NN	O	O
others	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
EBV	NN	O	O
can	NN	O	O
also	NN	O	O
infect	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
.	NN	O	O

Infection	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
linear	NN	O	B-DNA
EBV	NN	O	I-DNA
genome	NN	O	I-DNA
within	NN	O	O
8	NN	O	O
hr	NN	O	O
of	NN	O	O
infection	NN	O	O
.	NN	O	O

Circularization	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	B-DNA
genome	NN	O	I-DNA
was	NN	O	O
not	NN	O	O
detected	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
infection	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
where	NN	O	O
the	NN	O	O
genome	NN	O	O
can	NN	O	O
circularize	NN	O	O
within	NN	O	O
24	NN	O	O
hr	NN	O	O
of	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
appearance	NN	O	O
of	NN	O	O
the	NN	O	O
BamHI	NN	O	B-protein
ZLF-1	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
,	NN	O	O
ZEBRA	NN	O	B-protein
,	NN	O	O
by	NN	O	O
RT-PCR	NN	O	O
,	NN	O	O
was	NN	O	O
observed	NN	O	O
within	NN	O	O
8	NN	O	O
hr	NN	O	O
of	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
appearance	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
fusion	NN	O	B-protein
transcript	NN	O	I-protein
(	NN	O	O
RAZ	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
comprised	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
BZLF-1	NN	O	B-DNA
locus	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
adjacent	NN	O	O
BRLF-1	NN	O	B-DNA
locus	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
detected	NN	O	O
by	NN	O	O
RT-PCR	NN	O	O
.	NN	O	O

ZEBRA	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
also	NN	O	O
identified	NN	O	O
in	NN	O	O
infected	NN	O	O
thymocytes	NN	O	B-cell_type
by	NN	O	O
immunoprecipitation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
EBNA-1	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
infected	NN	O	O
thymocytes	NN	O	B-cell_type
was	NN	O	O
transcribed	NN	O	O
from	NN	O	O
the	NN	O	O
Fp	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
rather	NN	O	O
than	NN	O	O
from	NN	O	O
the	NN	O	O
Cp/Wp	NN	O	B-DNA
promoter	NN	O	I-DNA
which	NN	O	O
is	NN	O	O
used	NN	O	O
in	NN	O	O
latently	NN	O	B-cell_type
infected	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Transcripts	NN	O	O
encoding	NN	O	O
gp350/220	NN	O	B-protein
,	NN	O	O
the	NN	O	O
major	NN	O	B-protein
coat	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
EBV	NN	O	O
,	NN	O	O
were	NN	O	O
identified	NN	O	O
,	NN	O	O
but	NN	O	O
we	NN	O	O
did	NN	O	O
not	NN	O	O
find	NN	O	O
any	NN	O	O
evidence	NN	O	O
of	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
LMP-2A	NN	O	B-DNA
or	NN	O	O
EBER-1	NN	O	B-DNA
loci	NN	O	I-DNA
in	NN	O	O
infected	NN	O	O
thymocytes	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
de	NN	O	O
novo	NN	O	O
EBV	NN	O	O
infection	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
differs	NN	O	O
from	NN	O	O
infection	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
main	NN	O	O
difference	NN	O	O
is	NN	O	O
that	NN	O	O
with	NN	O	O
thymocytes	NN	O	B-cell_type
,	NN	O	O
no	NN	O	O
evidence	NN	O	O
could	NN	O	O
be	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
virus	NN	O	O
ever	NN	O	O
circularizes	NN	O	O
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
EBV	NN	O	O
remains	NN	O	O
in	NN	O	O
a	NN	O	O
linear	NN	O	O
configuration	NN	O	O
from	NN	O	O
which	NN	O	O
replicative	NN	O	B-DNA
genes	NN	O	I-DNA
are	NN	O	O
transcribed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
and	NN	O	O
purification	NN	O	O
of	NN	O	O
human	NN	O	B-protein
Stat	NN	O	I-protein
proteins	NN	O	I-protein
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
interleukin-2	NN	O	B-protein
.	NN	O	O

A	NN	O	O
key	NN	O	O
cytokine	NN	O	B-protein
induced	NN	O	O
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
is	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Following	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
the	NN	O	O
genes	NN	O	O
encoding	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
the	NN	O	O
various	NN	O	O
chains	NN	O	O
of	NN	O	O
its	NN	O	O
receptor	NN	O	O
are	NN	O	O
transcriptionally	NN	O	O
induced	NN	O	O
.	NN	O	O

In	NN	O	O
turn	NN	O	O
,	NN	O	O
secreted	NN	O	O
IL-2	NN	O	B-protein
serves	NN	O	O
to	NN	O	O
stimulate	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Several	NN	O	O
recent	NN	O	O
studies	NN	O	O
have	NN	O	O
implicated	NN	O	O
Jak	NN	O	O
kinases	NN	O	O
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Following	NN	O	O
this	NN	O	O
lead	NN	O	O
,	NN	O	O
we	NN	O	O
set	NN	O	O
out	NN	O	O
to	NN	O	O
identify	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
induced	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
observed	NN	O	O
to	NN	O	O
contain	NN	O	O
several	NN	O	O
IL-2	NN	O	B-protein
-inducible	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activities	NN	O	O
.	NN	O	O

Similar	NN	O	O
activities	NN	O	O
were	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
a	NN	O	O
transformed	NN	O	B-cell_line
human	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
termed	NN	O	O
YT	NN	O	B-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
purified	NN	O	O
these	NN	O	O
activities	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
principal	NN	O	O
IL-2-inducible	NN	O	B-protein
component	NN	O	I-protein
bears	NN	O	O
significant	NN	O	O
relatedness	NN	O	O
to	NN	O	O
a	NN	O	O
prolactin-induced	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
first	NN	O	O
identified	NN	O	O
in	NN	O	O
sheep	NN	O	B-cell_type
mammary	NN	O	I-cell_type
gland	NN	O	I-cell_type
tissue	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
,	NN	O	O
designated	NN	O	O
hStat5	NN	O	B-protein
,	NN	O	O
helps	NN	O	O
govern	NN	O	O
the	NN	O	O
biological	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

E2F-1	NN	O	B-protein
and	NN	O	O
a	NN	O	O
cyclin-like	NN	O	B-protein
DNA	NN	O	I-protein
repair	NN	O	I-protein
enzyme	NN	O	I-protein
,	NN	O	O
uracil-DNA	NN	O	B-protein
glycosylase	NN	O	I-protein
,	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
autoregulatory	NN	O	O
mechanism	NN	O	O
for	NN	O	O
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
cell	NN	O	B-protein
cycle-dependent	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
E2F-1	NN	O	B-protein
,	NN	O	O
regulates	NN	O	O
the	NN	O	O
cyclin	NN	O	B-protein
-like	NN	O	O
species	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
repair	NN	O	I-protein
enzyme	NN	O	I-protein
uracil-DNA	NN	O	B-DNA
glycosylase	NN	O	I-DNA
(	NN	O	I-DNA
UDG	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_line
osteosarcoma	NN	O	I-cell_line
(	NN	O	I-cell_line
Saos-2	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
,	NN	O	O
through	NN	O	O
the	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
UDG	NN	O	I-DNA
promoter	NN	O	I-DNA
sequences	NN	O	I-DNA
,	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
E2F-1	NN	O	B-protein
activates	NN	O	O
the	NN	O	O
UDG	NN	O	B-DNA
promoter	NN	O	I-DNA
through	NN	O	O
several	NN	O	O
E2F	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
major	NN	O	B-DNA
putative	NN	O	I-DNA
downstream	NN	O	I-DNA
site	NN	O	I-DNA
for	NN	O	O
E2F	NN	O	B-protein
,	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	B-DNA
exon	NN	O	I-DNA
,	NN	O	O
serves	NN	O	O
as	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
E2F-1/DP1	NN	O	B-protein
complex	NN	O	I-protein
binding	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
functional	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
cyclin-like	NN	O	B-protein
UDG	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
and	NN	O	O
E2F	NN	O	B-protein
.	NN	O	O

High	NN	O	O
levels	NN	O	O
of	NN	O	O
UDG	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
a	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assay	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
through	NN	O	O
elements	NN	O	B-DNA
specific	NN	O	O
for	NN	O	O
E2F	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
UDG	NN	O	B-protein
in	NN	O	O
Saos	NN	O	B-cell_line
2	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
observed	NN	O	O
to	NN	O	O
delay	NN	O	O
growth	NN	O	O
late	NN	O	O
in	NN	O	O
G1	NN	O	O
phase	NN	O	O
and	NN	O	O
transiently	NN	O	O
arrest	NN	O	O
these	NN	O	O
cells	NN	O	O
from	NN	O	O
progressing	NN	O	O
into	NN	O	O
the	NN	O	O
S	NN	O	O
phase	NN	O	O
.	NN	O	O

This	NN	O	O
hypothetical	NN	O	O
model	NN	O	O
integrates	NN	O	O
one	NN	O	O
mechanism	NN	O	O
of	NN	O	O
DNA	NN	O	O
repair	NN	O	O
with	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
control	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
likely	NN	O	O
through	NN	O	O
E2F	NN	O	B-protein
.	NN	O	O

This	NN	O	O
implicates	NN	O	O
E2F	NN	O	B-protein
as	NN	O	O
a	NN	O	O
multifunctional	NN	O	O
target	NN	O	O
for	NN	O	O
proteins	NN	O	O
and	NN	O	O
enzymes	NN	O	O
,	NN	O	O
possibly	NN	O	O
,	NN	O	O
responsive	NN	O	O
to	NN	O	O
DNA	NN	O	O
damage	NN	O	O
through	NN	O	O
the	NN	O	O
negative	NN	O	O
effect	NN	O	O
of	NN	O	O
UDG	NN	O	B-protein
on	NN	O	O
E2F	NN	O	B-protein
-mediated	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cellular	NN	O	O
and	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
NK	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
immunoregulatory	NN	O	B-protein
protein	NN	O	I-protein
produced	NN	O	O
predominantly	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
large	NN	O	B-cell_type
granular	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
LGL	NN	O	B-cell_type
)	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
different	NN	O	O
extracellular	NN	O	O
signals	NN	O	O
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
two	NN	O	O
interleukins	NN	O	B-protein
(	NN	O	O
ILs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
potent	NN	O	O
inducers	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
both	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
LGL	NN	O	B-cell_type
.	NN	O	O

Although	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
some	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
produce	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-2	NN	O	O
and	NN	O	O
IL-12	NN	O	O
stimulation	NN	O	O
,	NN	O	O
there	NN	O	O
has	NN	O	O
as	NN	O	O
yet	NN	O	O
been	NN	O	O
no	NN	O	O
report	NN	O	O
of	NN	O	O
a	NN	O	O
natural	NN	O	B-cell_line
killer	NN	O	I-cell_line
(	NN	O	I-cell_line
NK	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
that	NN	O	O
responds	NN	O	O
in	NN	O	O
a	NN	O	O
similar	NN	O	O
manner	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
present	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
NK3.3	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
responds	NN	O	O
to	NN	O	O
both	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
increases	NN	O	O
in	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
cytoplasmic	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
when	NN	O	O
used	NN	O	O
together	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
synergized	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
GM-CSF	NN	O	B-protein
and	NN	O	O
this	NN	O	O
synergy	NN	O	O
was	NN	O	O
attributed	NN	O	O
to	NN	O	O
an	NN	O	O
increased	NN	O	O
accumulation	NN	O	O
and	NN	O	O
stability	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-RNA
and	NN	O	I-RNA
GM-CSF	NN	O	I-RNA
mRNAs	NN	O	I-RNA
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
induction	NN	O	O
,	NN	O	O
five	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
,	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
,	NN	O	O
cycloheximide	NN	O	O
,	NN	O	O
genistein	NN	O	O
,	NN	O	O
and	NN	O	O
staurosporine	NN	O	O
A	NN	O	O
,	NN	O	O
were	NN	O	O
used	NN	O	O
in	NN	O	O
analyzing	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
on	NN	O	O
NK3.3	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
new	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
,	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-RNA
and	NN	O	I-RNA
GM-CSF	NN	O	I-RNA
cytoplasmic	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
IL-12	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-RNA
cytoplasmic	NN	O	I-RNA
mRNA	NN	O	I-RNA
appears	NN	O	O
to	NN	O	O
only	NN	O	O
partially	NN	O	O
depend	NN	O	O
on	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
both	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
and	NN	O	O
genistein	NN	O	O
,	NN	O	O
a	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
could	NN	O	O
suppress	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
but	NN	O	O
CsA	NN	O	O
was	NN	O	O
generally	NN	O	O
inactive	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
was	NN	O	O
observed	NN	O	O
that	NN	O	O
suppression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
these	NN	O	O
agents	NN	O	O
was	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
but	NN	O	O
not	NN	O	O
IL-12	NN	O	B-protein
induced	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
levels	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
complexes	NN	O	I-protein
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	O
and	NN	O	O
GM-CSF	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
may	NN	O	O
have	NN	O	O
distinct	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
GM-CSF	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
NK3.3	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
may	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
novel	NN	O	O
model	NN	O	O
for	NN	O	O
dissecting	NN	O	O
the	NN	O	O
biochemical	NN	O	O
and	NN	O	O
molecular	NN	O	O
events	NN	O	O
involved	NN	O	O
in	NN	O	O
these	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
functional	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
signaling	NN	O	O
pathway	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
p95vav	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
induces	NN	O	O
activation	NN	O	O
of	NN	O	O
multiple	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
numerous	NN	O	O
intracellular	NN	O	O
substrates	NN	O	O
.	NN	O	O

One	NN	O	O
substrate	NN	O	O
is	NN	O	O
p95vav	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
expressed	NN	O	O
exclusively	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
and	NN	O	I-cell_type
trophoblast	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
contains	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
structural	NN	O	B-protein
motifs	NN	O	I-protein
,	NN	O	O
including	NN	O	O
Src	NN	O	B-protein
homology	NN	O	I-protein
2	NN	O	I-protein
,	NN	O	I-protein
Src	NN	O	I-protein
homology	NN	O	I-protein
3	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
pleckstrin	NN	O	I-protein
homology	NN	O	I-protein
domains	NN	O	I-protein
and	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
guanine	NN	O	I-protein
nucleotide	NN	O	I-protein
exchange	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
p95vav	NN	O	B-protein
in	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
processes	NN	O	O
is	NN	O	O
unclear	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
p95vav	NN	O	B-protein
alone	NN	O	O
in	NN	O	O
Jurkat	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
leads	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
involved	NN	O	O
in	NN	O	O
interleukin-2	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
p95vav	NN	O	B-protein
synergizes	NN	O	O
with	NN	O	O
TCR	NN	O	B-protein
stimulation	NN	O	O
in	NN	O	O
inducing	NN	O	O
NFAT-	NN	O	O
and	NN	O	O
interleukin-2-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
a	NN	O	O
G-protein-coupled	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
not	NN	O	O
modulated	NN	O	O
by	NN	O	O
p95vav	NN	O	B-protein
overexpression	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
effect	NN	O	O
is	NN	O	O
specific	NN	O	O
to	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

Although	NN	O	O
removal	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	B-protein
67	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
of	NN	O	O
p95vav	NN	O	B-protein
activates	NN	O	O
its	NN	O	O
transforming	NN	O	O
potential	NN	O	O
in	NN	O	O
NIH	NN	O	B-cell_line
3T3	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
this	NN	O	O
region	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
its	NN	O	O
function	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
p95vav	NN	O	B-protein
-induced	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
not	NN	O	O
mimicked	NN	O	O
by	NN	O	O
Ras	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
though	NN	O	O
its	NN	O	O
function	NN	O	O
is	NN	O	O
dependent	NN	O	O
upon	NN	O	O
Ras	NN	O	B-protein
and	NN	O	O
Raf	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
activating	NN	O	O
function	NN	O	O
of	NN	O	O
p95vav	NN	O	B-protein
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
FK506	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
its	NN	O	O
activity	NN	O	O
also	NN	O	O
depends	NN	O	O
on	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

To	NN	O	O
further	NN	O	O
dissect	NN	O	O
p95vav	NN	O	B-protein
involvement	NN	O	O
in	NN	O	O
TCR	NN	O	B-protein
signaling	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
various	NN	O	O
Jurkat	NN	O	B-cell_line
mutants	NN	O	I-cell_line
deficient	NN	O	O
in	NN	O	O
TCR	NN	O	B-protein
signaling	NN	O	O
function	NN	O	O
or	NN	O	O
TCR	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
showed	NN	O	O
that	NN	O	O
an	NN	O	O
intact	NN	O	O
TCR	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
p95vav	NN	O	B-protein
to	NN	O	O
function	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
p95vav	NN	O	B-protein
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
influence	NN	O	O
TCR	NN	O	B-protein
-induced	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
or	NN	O	O
increases	NN	O	O
in	NN	O	O
cytoplasmic	NN	O	O
free	NN	O	O
calcium	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
p95vav	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
at	NN	O	O
an	NN	O	O
yet	NN	O	O
unidentified	NN	O	O
proximal	NN	O	O
position	NN	O	O
in	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
signaling	NN	O	O
cascade	NN	O	O
.	NN	O	O

-DOCSTART-	O

Positive	NN	O	O
and	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
promoter	NN	O	O
activity	NN	O	O
by	NN	O	O
AML1-related	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
PEBP2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
granulocyte-macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
GM-CSF	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
a	NN	O	O
consensus	NN	O	O
sequence	NN	O	O
for	NN	O	O
the	NN	O	O
polyomavirus	NN	O	B-protein
enhancer	NN	O	I-protein
binding-protein	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	I-protein
PEBP2	NN	O	I-protein
)	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
consists	NN	O	O
of	NN	O	O
alpha	NN	O	B-protein
and	NN	O	I-protein
beta	NN	O	I-protein
subunits	NN	O	I-protein
.	NN	O	O

There	NN	O	O
are	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
genes	NN	O	O
,	NN	O	O
alpha	NN	O	B-DNA
A	NN	O	I-DNA
and	NN	O	O
alpha	NN	O	B-DNA
B	NN	O	I-DNA
,	NN	O	O
encoding	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
subunit	NN	O	I-protein
.	NN	O	O

alpha	NN	O	B-DNA
B	NN	O	I-DNA
is	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
homologue	NN	O	I-DNA
of	NN	O	O
human	NN	O	B-DNA
AML1	NN	O	I-DNA
gene	NN	O	I-DNA
detected	NN	O	O
at	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
of	NN	O	O
t	NN	O	O
(	NN	O	O
8	NN	O	O
;	NN	O	O
21	NN	O	O
)	NN	O	O
and	NN	O	O
t	NN	O	O
(	NN	O	O
3	NN	O	O
;	NN	O	O
21	NN	O	O
)	NN	O	O
myeloid	NN	O	O
leukemias	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
alpha	NN	O	B-protein
A1	NN	O	I-protein
(	NN	O	O
an	NN	O	O
alpha	NN	O	B-protein
A-gene	NN	O	I-protein
product	NN	O	I-protein
)	NN	O	O
and	NN	O	O
alpha	NN	O	B-protein
B1	NN	O	I-protein
and	NN	O	O
alpha	NN	O	B-protein
B2	NN	O	I-protein
(	NN	O	O
two	NN	O	O
alpha	NN	O	B-protein
B-encoded	NN	O	I-protein
isomers	NN	O	I-protein
)	NN	O	O
for	NN	O	O
their	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

PEBP2	NN	O	B-protein
alpha	NN	O	I-protein
A1	NN	O	I-protein
,	NN	O	O
alpha	NN	O	B-protein
B1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
alpha	NN	O	B-protein
B2	NN	O	I-protein
proteins	NN	O	I-protein
bound	NN	O	O
the	NN	O	O
PEBP2	NN	O	B-DNA
site	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

PEBP2	NN	O	B-protein
alpha	NN	O	I-protein
A1	NN	O	I-protein
and	NN	O	O
alpha	NN	O	B-protein
B1	NN	O	I-protein
enhanced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter-driven	NN	O	I-DNA
reporter	NN	O	I-DNA
plasmid	NN	O	I-DNA
in	NN	O	O
unstimulated	NN	O	B-cell_line
and	NN	O	I-cell_line
12-O-tetradecanoylphorbol	NN	O	I-cell_line
13-acetate/phytohemagglutinin-stimulated	NN	O	I-cell_line
human	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
was	NN	O	O
suppressed	NN	O	O
by	NN	O	O
alpha	NN	O	B-protein
B2	NN	O	I-protein
.	NN	O	O

Coexpression	NN	O	O
of	NN	O	O
alpha	NN	O	B-protein
B1	NN	O	I-protein
and	NN	O	O
alpha	NN	O	B-protein
B2	NN	O	I-protein
showed	NN	O	O
that	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
could	NN	O	O
be	NN	O	O
determined	NN	O	O
by	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
B1	NN	O	I-protein
/alpha	NN	O	B-protein
B2	NN	O	I-protein
ratio	NN	O	O
.	NN	O	O

Jurkat	NN	O	O
cell	NN	O	O
extract	NN	O	O
contained	NN	O	O
PEBP2	NN	O	B-protein
site-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
that	NN	O	O
cross-reacted	NN	O	O
with	NN	O	O
antimouse	NN	O	B-protein
alpha	NN	O	I-protein
A1	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
indicated	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	O
PEBP2	NN	O	B-protein
alpha	NN	O	I-protein
A	NN	O	I-protein
,	NN	O	O
alpha	NN	O	B-DNA
B	NN	O	I-DNA
(	NN	O	O
AML1	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
and	NN	O	O
beta	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Although	NN	O	O
further	NN	O	O
studies	NN	O	O
are	NN	O	O
required	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
precise	NN	O	O
role	NN	O	O
of	NN	O	O
PEBP2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
the	NN	O	O
present	NN	O	O
findings	NN	O	O
suggested	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
the	NN	O	O
relative	NN	O	O
ratio	NN	O	O
of	NN	O	O
different	NN	O	O
PEBP2	NN	O	B-protein
isoforms	NN	O	I-protein
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

IFN-gamma	NN	O	B-protein
priming	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
enhances	NN	O	O
LPS-induced	NN	O	O
TNF	NN	O	B-protein
production	NN	O	O
by	NN	O	O
augmenting	NN	O	O
both	NN	O	O
transcription	NN	O	O
and	NN	O	O
MRNA	NN	O	O
stability	NN	O	O
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
/	NN	O	O
macrophages	NN	O	B-cell_type
by	NN	O	O
bacterial	NN	O	B-protein
endotoxin	NN	O	I-protein
or	NN	O	O
lipopolysaccharide	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
,	NN	O	O
highly	NN	O	O
regulated	NN	O	O
host	NN	O	O
defence	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
augmentation	NN	O	O
of	NN	O	O
LPS	NN	O	O
responses	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
priming	NN	O	O
,	NN	O	O
is	NN	O	O
well	NN	O	O
established	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
by	NN	O	O
which	NN	O	O
priming	NN	O	O
occurs	NN	O	O
is	NN	O	O
poorly	NN	O	O
defined	NN	O	O
.	NN	O	O

Using	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
induction	NN	O	O
as	NN	O	O
a	NN	O	O
model	NN	O	O
,	NN	O	O
experiments	NN	O	O
were	NN	O	O
designed	NN	O	O
to	NN	O	O
analyse	NN	O	O
in	NN	O	O
detail	NN	O	O
the	NN	O	O
priming	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
LPS	NN	O	O
response	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Priming	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
was	NN	O	O
primarily	NN	O	O
manifested	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
TNF	NN	O	B-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
.	NN	O	O

IFN-gamma	NN	O	B-protein
pre-treatment	NN	O	O
affected	NN	O	O
the	NN	O	O
magnitude	NN	O	O
rather	NN	O	O
than	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
the	NN	O	O
LPS	NN	O	O
response	NN	O	O
.	NN	O	O

Priming	NN	O	O
occurred	NN	O	O
after	NN	O	O
several	NN	O	O
hours	NN	O	O
of	NN	O	O
treatment	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
primed	NN	O	O
state	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
either	NN	O	O
IFN-gamma	NN	O	B-protein
or	NN	O	O
GM-CSF	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
M-CSF	NN	O	B-protein
.	NN	O	O

Primed	NN	O	O
monocytes	NN	O	B-cell_type
transcribed	NN	O	O
TNF	NN	O	B-RNA
mRNA	NN	O	I-RNA
at	NN	O	O
a	NN	O	O
higher	NN	O	O
rate	NN	O	O
than	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
monocytes	NN	O	B-cell_type
upon	NN	O	O
activation	NN	O	O
with	NN	O	O
LPS	NN	O	O
.	NN	O	O

The	NN	O	O
increased	NN	O	O
transcriptional	NN	O	O
rate	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
marked	NN	O	O
increase	NN	O	O
in	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
using	NN	O	O
a	NN	O	O
consensus	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
oligonucleotide	NN	O	I-DNA
.	NN	O	O

An	NN	O	O
additional	NN	O	O
significant	NN	O	O
finding	NN	O	O
was	NN	O	O
than	NN	O	O
TNF	NN	O	B-RNA
mRNA	NN	O	I-RNA
induced	NN	O	O
in	NN	O	O
primed	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
much	NN	O	O
more	NN	O	O
stable	NN	O	O
than	NN	O	O
in	NN	O	O
unprimed	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
T1/2	NN	O	O
increased	NN	O	O
6-8-fold	NN	O	O
)	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
increased	NN	O	O
mRNA	NN	O	B-RNA
stability	NN	O	O
,	NN	O	O
the	NN	O	O
duration	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
accumulation	NN	O	O
was	NN	O	O
longer	NN	O	O
following	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
in	NN	O	O
primed	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
being	NN	O	O
of	NN	O	O
greater	NN	O	O
magnitude	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
primed	NN	O	O
and	NN	O	O
unprimed	NN	O	O
cells	NN	O	O
possessed	NN	O	O
a	NN	O	O
differential	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
the	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
H-89	NN	O	O
.	NN	O	O

H-89	NN	O	O
substantially	NN	O	O
suppressed	NN	O	O
LPS-induced	NN	O	O
TNF	NN	O	B-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
in	NN	O	O
unprimed	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
primed	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
following	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

A	NN	O	O
Myc-associated	NN	O	B-DNA
zinc	NN	O	I-DNA
finger	NN	O	I-DNA
protein	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
is	NN	O	O
one	NN	O	O
of	NN	O	O
four	NN	O	O
important	NN	O	O
functional	NN	O	B-DNA
regions	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
developmental	NN	O	O
control	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
minimal	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
has	NN	O	O
four	NN	O	O
factor	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
each	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
full	NN	O	O
function	NN	O	O
.	NN	O	O

Using	NN	O	O
biochemical	NN	O	O
and	NN	O	O
mutagenesis	NN	O	O
analyses	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
that	NN	O	O
multiple	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
bind	NN	O	O
to	NN	O	O
these	NN	O	O
independent	NN	O	O
sites	NN	O	O
.	NN	O	O

We	NN	O	O
determined	NN	O	O
that	NN	O	O
an	NN	O	O
initiator-like	NN	O	B-DNA
sequence	NN	O	I-DNA
present	NN	O	O
at	NN	O	O
the	NN	O	O
cap	NN	O	B-DNA
site	NN	O	I-DNA
and	NN	O	O
an	NN	O	O
Ets	NN	O	B-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
full	NN	O	O
promoter	NN	O	B-DNA
function	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
Myc-associated	NN	O	B-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
MAZ	NN	O	B-protein
)	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
predominant	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
sites	NN	O	O
.	NN	O	O

This	NN	O	O
last	NN	O	O
site	NN	O	O
closely	NN	O	O
resembles	NN	O	O
the	NN	O	O
ME1a1	NN	O	B-DNA
G3AG4AG3	NN	O	I-DNA
motif	NN	O	I-DNA
previously	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
critical	NN	O	O
element	NN	O	O
in	NN	O	O
the	NN	O	O
P2	NN	O	B-DNA
promoter	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
c-myc	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
believe	NN	O	O
that	NN	O	O
the	NN	O	O
MAZ	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
also	NN	O	O
likely	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
developmental	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Erythropoietin	NN	O	B-protein
stimulates	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
TAL1/SCL	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
its	NN	O	O
protein	NN	O	B-protein
products	NN	O	I-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
TAL1	NN	O	B-DNA
(	NN	O	I-DNA
or	NN	O	I-DNA
SCL	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
originally	NN	O	O
identified	NN	O	O
through	NN	O	O
its	NN	O	O
involvement	NN	O	O
by	NN	O	O
a	NN	O	O
recurrent	NN	O	O
chromosomal	NN	O	O
translocation	NN	O	O
,	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
frequent	NN	O	O
molecular	NN	O	O
lesion	NN	O	O
recognized	NN	O	O
in	NN	O	O
T-cell	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

The	NN	O	O
protein	NN	O	B-protein
products	NN	O	I-protein
of	NN	O	O
this	NN	O	O
gene	NN	O	O
contain	NN	O	O
the	NN	O	O
basic-helix-loop-helix	NN	O	B-protein
motif	NN	O	I-protein
characteristic	NN	O	O
of	NN	O	O
a	NN	O	O
large	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
canonical	NN	O	B-DNA
DNA	NN	O	I-DNA
sequence	NN	O	I-DNA
CANNTG	NN	O	I-DNA
as	NN	O	O
protein	NN	O	B-protein
heterodimers	NN	O	I-protein
.	NN	O	O

TAL1	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
chemical-induced	NN	O	B-cell_line
erythroleukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
in	NN	O	O
vivo	NN	O	O
suggested	NN	O	O
the	NN	O	O
gene	NN	O	O
might	NN	O	O
regulate	NN	O	O
aspects	NN	O	O
of	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
terminal	NN	O	O
events	NN	O	O
of	NN	O	O
erythropoiesis	NN	O	O
are	NN	O	O
controlled	NN	O	O
by	NN	O	O
the	NN	O	O
glycoprotein	NN	O	B-protein
hormone	NN	O	I-protein
erythropoietin	NN	O	I-protein
(	NN	O	O
Epo	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
the	NN	O	O
expression	NN	O	O
or	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
TAL1	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
its	NN	O	O
protein	NN	O	B-protein
products	NN	O	I-protein
were	NN	O	O
affected	NN	O	O
by	NN	O	O
Epo	NN	O	B-protein
in	NN	O	O
splenic	NN	O	B-cell_type
erythroblasts	NN	O	I-cell_type
from	NN	O	O
mice	NN	O	O
infected	NN	O	O
with	NN	O	O
an	NN	O	O
anemia-inducing	NN	O	O
strain	NN	O	O
of	NN	O	O
Friend	NN	O	O
virus	NN	O	O
(	NN	O	O
FVA	NN	O	O
cells	NN	O	O
)	NN	O	O
.	NN	O	O

Epo	NN	O	B-protein
elicited	NN	O	O
a	NN	O	O
rapid	NN	O	O
,	NN	O	O
dose-related	NN	O	O
increase	NN	O	O
in	NN	O	O
TAL1	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
increasing	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
and	NN	O	O
stabilizing	NN	O	O
one	NN	O	O
of	NN	O	O
its	NN	O	O
mRNAs	NN	O	B-RNA
.	NN	O	O

An	NN	O	O
Epo	NN	O	B-protein
-inducible	NN	O	O
TAL1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
FVA	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
that	NN	O	O
subsequently	NN	O	O
decayed	NN	O	O
despite	NN	O	O
accumulating	NN	O	O
mRNA	NN	O	B-RNA
and	NN	O	O
protein	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
associated	NN	O	O
temporally	NN	O	O
with	NN	O	O
Epo	NN	O	B-protein
-induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
TAL1	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
Epo	NN	O	B-protein
acts	NN	O	O
at	NN	O	O
both	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
posttranscriptional	NN	O	O
levels	NN	O	O
on	NN	O	O
the	NN	O	O
TAL1	NN	O	B-protein
locus	NN	O	O
in	NN	O	O
Friend	NN	O	B-cell_line
virus-induced	NN	O	I-cell_line
erythroblasts	NN	O	I-cell_line
and	NN	O	O
establish	NN	O	O
a	NN	O	O
link	NN	O	O
between	NN	O	O
Epo	NN	O	B-protein
signaling	NN	O	O
mechanisms	NN	O	O
and	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
diverse	NN	O	B-cell_type
cell	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Nonradioactive	NN	O	O
quantification	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
U937	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
a	NN	O	O
method	NN	O	O
for	NN	O	O
relative	NN	O	O
quantification	NN	O	O
of	NN	O	O
specific	NN	O	O
mRNA	NN	O	B-RNA
using	NN	O	O
a	NN	O	O
nonradioactive	NN	O	O
assay	NN	O	O
based	NN	O	O
on	NN	O	O
DNA	NN	O	O
strand	NN	O	O
competition	NN	O	O
between	NN	O	O
identical	NN	O	O
sequences	NN	O	O
of	NN	O	O
biotin-	NN	O	O
and	NN	O	O
fluorescein-labeled	NN	O	O
amplicon	NN	O	O
(	NN	O	O
probe	NN	O	O
)	NN	O	O
and	NN	O	O
unlabeled	NN	O	O
amplicon	NN	O	O
(	NN	O	O
target	NN	O	O
)	NN	O	O
during	NN	O	O
hybridization	NN	O	O
.	NN	O	O

As	NN	O	O
the	NN	O	O
target	NN	O	O
quantity	NN	O	O
increased	NN	O	O
,	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
double-labeled	NN	O	O
probe	NN	O	O
decreased	NN	O	O
in	NN	O	O
accordance	NN	O	O
with	NN	O	O
the	NN	O	O
mass	NN	O	O
action	NN	O	O
law	NN	O	O
.	NN	O	O

This	NN	O	O
technique	NN	O	O
was	NN	O	O
successfully	NN	O	O
applied	NN	O	O
to	NN	O	O
evaluate	NN	O	O
differences	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
before	NN	O	O
and	NN	O	O
after	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
potent	NN	O	O
differentiation	NN	O	O
inducers	NN	O	O
:	NN	O	O
12-O-tetradecanoylphorbol	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
and	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D2	NN	O	O
(	NN	O	O
VD	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
TPA	NN	O	O
treatment	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
specific	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
while	NN	O	O
no	NN	O	O
enhanced	NN	O	O
GR	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
perceived	NN	O	O
with	NN	O	O
RA/VD	NN	O	O
treatment	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
Sp1	NN	O	B-protein
phosphorylation	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-DNA
B-independent	NN	O	I-DNA
HIV	NN	O	I-DNA
promoter	NN	O	I-DNA
domain	NN	O	O
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
purely	NN	O	O
enhancer-dependent	NN	O	O
effect	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
such	NN	O	O
as	NN	O	O
TNF	NN	O	B-protein
on	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
(	NN	O	O
OKA	NN	O	O
)	NN	O	O
activates	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
through	NN	O	O
both	NN	O	O
the	NN	O	O
enhancer	NN	O	O
,	NN	O	O
responding	NN	O	O
to	NN	O	O
the	NN	O	O
factor	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
domain	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
inducibility	NN	O	O
of	NN	O	O
HIV	NN	O	B-DNA
LTR-driven	NN	O	I-DNA
luciferase	NN	O	I-DNA
expression	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
lymphoblastoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
OKA	NN	O	O
depended	NN	O	O
on	NN	O	O
both	NN	O	O
functional	NN	O	B-DNA
Sp1	NN	O	I-DNA
binding	NN	O	I-DNA
elements	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
to	NN	O	O
bind	NN	O	O
the	NN	O	O
protein	NN	O	O
TBP	NN	O	B-protein
.	NN	O	O

In	NN	O	O
both	NN	O	O
transformed	NN	O	B-cell_type
and	NN	O	I-cell_type
normal	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
OKA	NN	O	O
stimulation	NN	O	O
induced	NN	O	O
intense	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
Sp1	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
a	NN	O	O
property	NN	O	O
of	NN	O	O
OKA	NN	O	O
not	NN	O	O
shared	NN	O	O
by	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
,	NN	O	O
or	NN	O	O
PHA	NN	O	B-protein
and	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
.	NN	O	O

Responsiveness	NN	O	O
of	NN	O	O
LTR	NN	O	B-DNA
constructs	NN	O	I-DNA
deleted	NN	O	O
of	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
elements	NN	O	I-DNA
to	NN	O	O
HIV	NN	O	B-protein
Tat	NN	O	I-protein
expression	NN	O	O
was	NN	O	O
increased	NN	O	O
upon	NN	O	O
OKA	NN	O	O
but	NN	O	O
not	NN	O	O
TNF	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
SP1	NN	O	B-protein
phosphorylation	NN	O	O
induced	NN	O	O
by	NN	O	O
OKA	NN	O	O
,	NN	O	O
a	NN	O	O
selective	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
the	NN	O	O
serine-threonine	NN	O	B-protein
phosphatase	NN	O	I-protein
PP2A	NN	O	B-protein
,	NN	O	O
facilitates	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
complex	NN	O	I-protein
involving	NN	O	O
general	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
HIV	NN	O	B-protein
Tat	NN	O	I-protein
,	NN	O	O
and	NN	O	O
Sp1	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
formation	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
would	NN	O	O
increase	NN	O	O
,	NN	O	O
independently	NN	O	O
of	NN	O	O
an	NN	O	O
in	NN	O	O
synergy	NN	O	O
with	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
the	NN	O	O
low	NN	O	O
basal	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
observed	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
shared	NN	O	B-protein
receptor	NN	O	I-protein
motifs	NN	O	I-protein
and	NN	O	O
common	NN	O	B-protein
Stat	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
pleiotropy	NN	O	O
and	NN	O	O
redundancy	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-7	NN	O	B-protein
,	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-15	NN	O	B-protein
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
molecular	NN	O	O
bases	NN	O	O
for	NN	O	O
cytokine	NN	O	O
redundancy	NN	O	O
and	NN	O	O
pleiotropy	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
compared	NN	O	O
the	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
activated	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
PBLs	NN	O	B-cell_type
)	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
with	NN	O	O
shared	NN	O	O
and	NN	O	O
distinct	NN	O	O
actions	NN	O	O
.	NN	O	O

Interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
rapidly	NN	O	O
activated	NN	O	O
Stat5	NN	O	B-protein
in	NN	O	O
fresh	NN	O	O
PBL	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
Stat3	NN	O	B-protein
and	NN	O	O
Stat5	NN	O	B-protein
in	NN	O	O
preactivated	NN	O	B-cell_type
PBL	NN	O	I-cell_type
.	NN	O	O

IL-7	NN	O	B-protein
and	NN	O	O
IL-15	NN	O	B-protein
induced	NN	O	O
the	NN	O	O
same	NN	O	O
complexes	NN	O	O
as	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
a	NN	O	O
feature	NN	O	O
explained	NN	O	O
by	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
similar	NN	O	O
tyrosine-phosphorylated	NN	O	B-protein
motifs	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
IL-7R	NN	O	B-protein
that	NN	O	O
can	NN	O	O
serve	NN	O	O
as	NN	O	O
docking	NN	O	O
sites	NN	O	O
for	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

IL-13	NN	O	B-protein
Induced	NN	O	O
the	NN	O	O
same	NN	O	O
complexes	NN	O	O
as	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
a	NN	O	O
finding	NN	O	O
explained	NN	O	O
by	NN	O	O
our	NN	O	O
studies	NN	O	O
implicating	NN	O	O
IL-4R	NN	O	B-protein
as	NN	O	O
a	NN	O	O
shared	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
receptors	NN	O	B-protein
.	NN	O	O

These	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
single	NN	O	O
cytokine	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
different	NN	O	O
combinations	NN	O	O
of	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
under	NN	O	O
different	NN	O	O
physiological	NN	O	O
conditions	NN	O	O
,	NN	O	O
and	NN	O	O
also	NN	O	O
indicate	NN	O	O
two	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
distinct	NN	O	O
cytokines	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
the	NN	O	O
same	NN	O	O
Stat	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Control	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
proteolysis	NN	O	O
by	NN	O	O
site-specific	NN	O	O
,	NN	O	O
signal-induced	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
inhibits	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
retaining	NN	O	O
it	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

Various	NN	O	O
stimuli	NN	O	O
,	NN	O	O
typically	NN	O	O
those	NN	O	O
associated	NN	O	O
with	NN	O	O
stress	NN	O	O
or	NN	O	O
pathogens	NN	O	O
,	NN	O	O
rapidly	NN	O	O
inactivate	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
.	NN	O	O

This	NN	O	O
liberates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
to	NN	O	O
translocate	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
and	NN	O	O
initiate	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
important	NN	O	O
for	NN	O	O
the	NN	O	O
defense	NN	O	O
of	NN	O	O
the	NN	O	O
organism	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
correlates	NN	O	O
with	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
and	NN	O	O
requires	NN	O	O
the	NN	O	O
proteolysis	NN	O	O
of	NN	O	O
this	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

When	NN	O	O
either	NN	O	O
serine-32	NN	O	O
or	NN	O	O
serine-36	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
was	NN	O	O
mutated	NN	O	O
,	NN	O	O
the	NN	O	O
protein	NN	O	O
did	NN	O	O
not	NN	O	O
undergo	NN	O	O
signal-induced	NN	O	O
phosphorylation	NN	O	O
or	NN	O	O
degradation	NN	O	O
,	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
could	NN	O	O
not	NN	O	O
be	NN	O	O
activated	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
phosphorylation	NN	O	O
at	NN	O	O
one	NN	O	O
or	NN	O	O
both	NN	O	O
of	NN	O	O
these	NN	O	O
residues	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
erythropoietin	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
proteins	NN	O	O
binding	NN	O	O
to	NN	O	O
GATA-1	NN	O	B-DNA
and	NN	O	I-DNA
Sp1	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

Erythropoietin	NN	O	B-protein
(	NN	O	O
Epo	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
primary	NN	O	O
regulator	NN	O	O
of	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
acts	NN	O	O
by	NN	O	O
binding	NN	O	O
to	NN	O	O
a	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
EpoR	NN	O	B-protein
)	NN	O	O
on	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
used	NN	O	O
deletion	NN	O	O
analysis	NN	O	O
and	NN	O	O
transfection	NN	O	O
assays	NN	O	O
with	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
to	NN	O	O
examine	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
control	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
EpoR	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
most	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
activity	NN	O	O
was	NN	O	O
contained	NN	O	O
in	NN	O	O
a	NN	O	O
150	NN	O	B-DNA
bp	NN	O	I-DNA
promoter	NN	O	I-DNA
fragment	NN	O	O
with	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP2	NN	O	B-protein
,	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
erythroid-specific	NN	O	B-protein
GATA-1	NN	O	I-protein
.	NN	O	O

The	NN	O	O
150	NN	O	B-DNA
bp	NN	O	I-DNA
hEpoR	NN	O	I-DNA
promoter	NN	O	I-DNA
exhibited	NN	O	O
high	NN	O	O
and	NN	O	O
low	NN	O	O
activity	NN	O	O
in	NN	O	O
erythroid	NN	O	O
OCIM1	NN	O	B-cell_line
and	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
reflecting	NN	O	O
the	NN	O	O
high	NN	O	O
and	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
constitutive	NN	O	O
hEpoR	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
GATA-1	NN	O	B-DNA
and	NN	O	I-DNA
Sp1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
this	NN	O	O
promoter	NN	O	O
lacking	NN	O	O
a	NN	O	O
TATA	NN	O	B-DNA
sequence	NN	O	I-DNA
were	NN	O	O
necessary	NN	O	O
for	NN	O	O
a	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
activation	NN	O	O
.	NN	O	O

Protein-DNA	NN	O	O
binding	NN	O	O
studies	NN	O	O
suggested	NN	O	O
that	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
two	NN	O	O
other	NN	O	O
CCGCCC	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
from	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
non-erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
could	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
Sp1	NN	O	B-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

By	NN	O	O
increasing	NN	O	O
GATA-1	NN	O	B-protein
levels	NN	O	O
via	NN	O	O
co-transfection	NN	O	O
,	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
transactivate	NN	O	O
the	NN	O	O
hEpoR	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
non-erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
highly	NN	O	O
active	NN	O	O
OCIM1	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
although	NN	O	O
GATA-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
were	NN	O	O
comparable	NN	O	O
in	NN	O	O
OCIM1	NN	O	B-cell_line
and	NN	O	O
K562	NN	O	B-cell_line
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
when	NN	O	O
we	NN	O	O
mutated	NN	O	O
the	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
marked	NN	O	O
decrease	NN	O	O
in	NN	O	O
hEpoR	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
,	NN	O	O
we	NN	O	O
could	NN	O	O
restore	NN	O	O
transactivation	NN	O	O
by	NN	O	O
increasing	NN	O	O
GATA-1	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
OCIM1	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
while	NN	O	O
GATA-1	NN	O	B-protein
can	NN	O	O
transactivate	NN	O	O
the	NN	O	O
EpoR	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
hEpoR	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
does	NN	O	O
not	NN	O	O
depend	NN	O	O
on	NN	O	O
GATA-1	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
hEpoR	NN	O	B-protein
transcription	NN	O	O
activity	NN	O	O
depends	NN	O	O
on	NN	O	O
coordination	NN	O	O
between	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
GATA-1	NN	O	B-protein
with	NN	O	O
other	NN	O	O
cell-specific	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
possibly	NN	O	O
other	NN	O	O
Sp1-like	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
to	NN	O	O
provide	NN	O	O
high	NN	O	O
level	NN	O	O
,	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Overexpression	NN	O	O
of	NN	O	O
DR-nm23	NN	O	B-protein
,	NN	O	O
a	NN	O	O
protein	NN	O	O
encoded	NN	O	O
by	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
nm23	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
,	NN	O	O
inhibits	NN	O	O
granulocyte	NN	O	B-cell_type
differentiation	NN	O	O
and	NN	O	O
induces	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
32Dc13	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
evolves	NN	O	O
in	NN	O	O
two	NN	O	O
clinically	NN	O	O
distinct	NN	O	O
stages	NN	O	O
:	NN	O	O
a	NN	O	O
chronic	NN	O	O
and	NN	O	O
a	NN	O	O
blast	NN	O	O
crisis	NN	O	O
phase	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
changes	NN	O	O
associated	NN	O	O
with	NN	O	O
chronic	NN	O	O
phase	NN	O	O
to	NN	O	O
blast	NN	O	O
crisis	NN	O	O
transition	NN	O	O
are	NN	O	O
largely	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
,	NN	O	O
DR-nm23	NN	O	B-protein
,	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
blast-crisis	NN	O	B-DNA
cDNA	NN	O	I-DNA
library	NN	O	I-DNA
,	NN	O	I-DNA
which	NN	O	O
has	NN	O	O
approximately	NN	O	O
70	NN	O	O
%	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
to	NN	O	O
the	NN	O	O
putative	NN	O	B-DNA
metastatic	NN	O	I-DNA
suppressor	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
nm23-H1	NN	O	B-DNA
and	NN	O	O
nm23-H2	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
deduced	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
to	NN	O	O
the	NN	O	O
proteins	NN	O	O
encoded	NN	O	O
by	NN	O	O
these	NN	O	O
two	NN	O	O
latter	NN	O	O
genes	NN	O	O
is	NN	O	O
approximately	NN	O	O
65	NN	O	O
%	NN	O	O
and	NN	O	O
includes	NN	O	O
domains	NN	O	O
and	NN	O	O
amino	NN	O	O
acid	NN	O	O
residues	NN	O	O
(	NN	O	O
the	NN	O	O
leucine	NN	O	B-protein
zipper-like	NN	O	I-protein
and	NN	O	O
the	NN	O	O
RGD	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
a	NN	O	O
serine	NN	O	O
and	NN	O	O
a	NN	O	O
histidine	NN	O	O
residue	NN	O	O
in	NN	O	O
the	NN	O	O
NH2-	NN	O	B-protein
and	NN	O	O
in	NN	O	O
the	NN	O	O
COOH-terminal	NN	O	B-protein
portion	NN	O	I-protein
of	NN	O	O
the	NN	O	O
protein	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
postulated	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
nm23	NN	O	B-protein
function	NN	O	O
.	NN	O	O

DR-nm23	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
preferentially	NN	O	O
expressed	NN	O	O
at	NN	O	O
early	NN	O	O
stages	NN	O	O
of	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
highly	NN	O	B-cell_line
purified	NN	O	I-cell_line
CD34+	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Its	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
myeloid	NN	O	B-cell_line
precursor	NN	O	I-cell_line
32Dc13	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
is	NN	O	O
growth-factor	NN	O	O
dependent	NN	O	O
for	NN	O	O
both	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
,	NN	O	O
results	NN	O	O
in	NN	O	O
inhibition	NN	O	O
of	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
causes	NN	O	O
apoptotic	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
DR-nm23	NN	O	B-protein
in	NN	O	O
normal	NN	O	O
hematopoiesis	NN	O	O
and	NN	O	O
raise	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
its	NN	O	O
overexpression	NN	O	O
contributes	NN	O	O
to	NN	O	O
differentiation	NN	O	O
arrest	NN	O	O
,	NN	O	O
a	NN	O	O
feature	NN	O	O
of	NN	O	O
blastic	NN	O	O
transformation	NN	O	O
in	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
interferon-gamma	NN	O	B-DNA
activation	NN	O	I-DNA
sequence	NN	O	I-DNA
mediates	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
IgG	NN	O	B-DNA
Fc	NN	O	I-DNA
receptor	NN	O	I-DNA
type	NN	O	I-DNA
IC	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
interferon-gamma	NN	O	B-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
IgG	NN	O	B-protein
Fc	NN	O	I-protein
receptor	NN	O	I-protein
type	NN	O	I-protein
I	NN	O	I-protein
(	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RI	NN	O	I-protein
)	NN	O	O
on	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
dramatically	NN	O	O
increased	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RI	NN	O	I-protein
transcript	NN	O	O
levels	NN	O	O
in	NN	O	O
monoblast-like	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
elevated	NN	O	O
within	NN	O	O
3	NN	O	O
hr	NN	O	O
and	NN	O	O
peaked	NN	O	O
12	NN	O	O
hr	NN	O	O
after	NN	O	O
exposure	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
with	NN	O	O
IFN-gamma	NN	O	B-protein
for	NN	O	O
9	NN	O	O
hr	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cycloheximide	NN	O	O
led	NN	O	O
to	NN	O	O
super-induction	NN	O	O
of	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RI	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
run-on	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RI	NN	O	I-protein
transcription	NN	O	O
was	NN	O	O
increased	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

Genomic	NN	O	O
sequence	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
Fc	NN	O	B-DNA
gamma	NN	O	I-DNA
RIC	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
cloned	NN	O	O
and	NN	O	O
subjected	NN	O	O
to	NN	O	O
primer	NN	O	O
extension	NN	O	O
analysis	NN	O	O
,	NN	O	O
which	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
single	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
without	NN	O	O
a	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
.	NN	O	O

Transient	NN	O	O
transfections	NN	O	O
of	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
containing	NN	O	O
various	NN	O	O
Fc	NN	O	B-DNA
gamma	NN	O	I-DNA
RIC	NN	O	I-DNA
promoter	NN	O	I-DNA
sequences	NN	O	I-DNA
into	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
revealed	NN	O	O
that	NN	O	O
a	NN	O	O
20-bp	NN	O	B-DNA
region	NN	O	I-DNA
surrounding	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
-7	NN	O	O
to	NN	O	O
+13	NN	O	O
)	NN	O	O
was	NN	O	O
capable	NN	O	O
of	NN	O	O
mediating	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
and	NN	O	O
that	NN	O	O
an	NN	O	O
IFN-gamma	NN	O	B-DNA
responsive	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
GIRE	NN	O	B-DNA
)	NN	O	O
was	NN	O	O
present	NN	O	O
within	NN	O	O
74	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
17-bp	NN	O	B-DNA
sequence	NN	O	I-DNA
between	NN	O	O
positions	NN	O	O
-51	NN	O	O
and	NN	O	O
-35	NN	O	O
conferred	NN	O	O
IFN-gamma	NN	O	B-protein
responsiveness	NN	O	O
on	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Double-stranded	NN	O	B-DNA
GIRE	NN	O	I-DNA
sequence	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
a	NN	O	O
scrambled	NN	O	B-DNA
sequence	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
specifically	NN	O	O
bound	NN	O	O
by	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
from	NN	O	O
IFN-gamma	NN	O	B-cell_line
treated	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Gel	NN	O	O
shift	NN	O	O
experiments	NN	O	O
further	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
protein	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
Fc	NN	O	B-DNA
gamma	NN	O	I-DNA
RIC	NN	O	I-DNA
GIRE	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
treatment	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
Fc	NN	O	B-DNA
gamma	NN	O	I-DNA
RIC	NN	O	I-DNA
GIRE	NN	O	I-DNA
is	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-DNA
activation	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
GAS	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
guanylate	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
to	NN	O	O
X	NN	O	B-DNA
box	NN	O	I-DNA
elements	NN	O	I-DNA
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
MHC	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
Fc	NN	O	B-DNA
gamma	NN	O	I-DNA
RIC	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
IFN-gamma	NN	O	B-protein
involves	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
to	NN	O	O
a	NN	O	O
17-bp	NN	O	B-DNA
GAS	NN	O	I-DNA
homology	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Constitutively	NN	O	O
activated	NN	O	O
Jak	NN	O	B-protein
-STAT	NN	O	B-protein
pathway	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
transformed	NN	O	O
with	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

Human	NN	O	O
T	NN	O	O
cell	NN	O	O
lymphotropic	NN	O	O
virus	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
is	NN	O	O
the	NN	O	O
etiological	NN	O	O
agent	NN	O	O
for	NN	O	O
adult	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
and	NN	O	O
tropical	NN	O	O
spastic	NN	O	O
paraparesis	NN	O	O
(	NN	O	O
also	NN	O	O
termed	NN	O	O
HTLV-I-associated	NN	O	O
myelopathy	NN	O	O
)	NN	O	O
.	NN	O	O

HTLV-I-infected	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
exhibit	NN	O	O
an	NN	O	O
initial	NN	O	O
phase	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
-dependent	NN	O	O
growth	NN	O	O
;	NN	O	O
over	NN	O	O
time	NN	O	O
,	NN	O	O
by	NN	O	O
an	NN	O	O
unknown	NN	O	O
mechanism	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
become	NN	O	O
IL-2	NN	O	B-protein
-independent	NN	O	O
.	NN	O	O

Whereas	NN	O	O
the	NN	O	O
Jak	NN	O	B-protein
kinases	NN	O	I-protein
Jak1	NN	O	B-protein
and	NN	O	O
Jak3	NN	O	B-protein
and	NN	O	O
the	NN	O	O
signal	NN	O	O
transducer	NN	O	O
and	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
proteins	NN	O	I-protein
Stat3	NN	O	B-protein
and	NN	O	O
Stat5	NN	O	B-protein
are	NN	O	O
activated	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
this	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
was	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
in	NN	O	O
HTLV-I-transformed	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
HTLV-I-infected	NN	O	B-cell_line
cord	NN	O	I-cell_line
blood	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
transition	NN	O	O
from	NN	O	O
IL-2	NN	O	B-protein
-dependent	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
-independent	NN	O	O
growth	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
acquisition	NN	O	O
of	NN	O	O
a	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
Jak	NN	O	B-protein
-STAT	NN	O	B-protein
pathway	NN	O	O
,	NN	O	O
which	NN	O	O
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
pathway	NN	O	O
participates	NN	O	O
in	NN	O	O
HTLV-I-mediated	NN	O	O
T	NN	O	O
cell	NN	O	O
transformation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nitric	NN	O	O
oxide	NN	O	O
decreases	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
endothelial	NN	O	O
activation	NN	O	O
.	NN	O	O

Nitric	NN	O	O
oxide	NN	O	O
selectively	NN	O	O
reduces	NN	O	O
endothelial	NN	O	O
expression	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
and	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

To	NN	O	O
test	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
(	NN	O	O
NO	NN	O	O
)	NN	O	O
limits	NN	O	O
endothelial	NN	O	O
activation	NN	O	O
,	NN	O	O
we	NN	O	O
treated	NN	O	O
cytokine-stimulated	NN	O	B-cell_line
human	NN	O	I-cell_line
saphenous	NN	O	I-cell_line
vein	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
several	NN	O	O
NO	NN	O	O
donors	NN	O	O
and	NN	O	O
assessed	NN	O	O
their	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
NO	NN	O	O
inhibited	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
alpha-stimulated	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
35-55	NN	O	O
%	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
cell	NN	O	O
surface	NN	O	O
enzyme	NN	O	O
immunoassays	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
was	NN	O	O
paralleled	NN	O	O
by	NN	O	O
reduced	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
endothelial	NN	O	O
monolayers	NN	O	O
in	NN	O	O
nonstatic	NN	O	O
assays	NN	O	O
,	NN	O	O
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
cGMP	NN	O	O
analogues	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
quantitatively	NN	O	O
similar	NN	O	O
after	NN	O	O
stimulation	NN	O	O
by	NN	O	O
either	NN	O	O
IL-1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
,	NN	O	O
or	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
.	NN	O	O

NO	NN	O	O
also	NN	O	O
decreased	NN	O	O
the	NN	O	O
endothelial	NN	O	O
expression	NN	O	O
of	NN	O	O
other	NN	O	O
leukocyte	NN	O	B-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
(	NN	O	O
E-selectin	NN	O	B-protein
and	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
)	NN	O	O
and	NN	O	O
secretable	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
IL-8	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
endogenous	NN	O	O
NO	NN	O	O
production	NN	O	O
by	NN	O	O
L-N-monomethyl-arginine	NN	O	O
also	NN	O	O
induced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
augment	NN	O	O
cytokine-induced	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
run-on	NN	O	O
assays	NN	O	O
,	NN	O	O
transfection	NN	O	O
studies	NN	O	O
using	NN	O	O
various	NN	O	O
VCAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
indicated	NN	O	O
that	NN	O	O
NO	NN	O	O
represses	NN	O	O
VCAM-1	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
NO	NN	O	O
's	NN	O	O
ability	NN	O	O
to	NN	O	O
limit	NN	O	O
endothelial	NN	O	O
activation	NN	O	O
and	NN	O	O
inhibit	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
some	NN	O	O
of	NN	O	O
its	NN	O	O
antiatherogenic	NN	O	O
and	NN	O	O
antiinflammatory	NN	O	O
properties	NN	O	O
within	NN	O	O
the	NN	O	O
vessel	NN	O	O
wall	NN	O	O
.	NN	O	O

-DOCSTART-	O

MIP1	NN	O	B-protein
alpha	NN	O	I-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
MNP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
novel	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
hematopoietic	NN	O	O
cells	NN	O	O
that	NN	O	O
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
MIP-1	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Murine	NN	O	B-protein
macrophage	NN	O	I-protein
inflammatory	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
MIP-1	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
and	NN	O	O
its	NN	O	O
human	NN	O	B-protein
equivalent	NN	O	I-protein
(	NN	O	O
GOS19	NN	O	B-protein
,	NN	O	O
LD78	NN	O	B-protein
,	NN	O	O
or	NN	O	O
AT464	NN	O	B-protein
)	NN	O	O
are	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
-C-C	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
low-molecular-weight	NN	O	B-protein
chemokines	NN	O	I-protein
.	NN	O	O

Secreted	NN	O	O
from	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
bone	NN	O	O
marrow-derived	NN	O	B-protein
MIP-1	NN	O	I-protein
alpha/GOS19	NN	O	I-protein
inhibits	NN	O	O
primitive	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
homeostatic	NN	O	O
control	NN	O	O
of	NN	O	O
stem	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
induces	NN	O	O
chemotaxis	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
important	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
which	NN	O	O
control	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
MIP-1	NN	O	B-protein
alpha	NN	O	I-protein
/GOS19	NN	O	B-protein
.	NN	O	O

Previous	NN	O	O
work	NN	O	O
has	NN	O	O
shown	NN	O	O
that	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
widely	NN	O	O
expressed	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
the	NN	O	O
ICK-1	NN	O	B-protein
family	NN	O	I-protein
)	NN	O	O
affect	NN	O	O
the	NN	O	O
GOS19	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

One	NN	O	O
member	NN	O	O
,	NN	O	O
ICK-1A	NN	O	B-protein
,	NN	O	O
behaves	NN	O	O
as	NN	O	O
a	NN	O	O
strong	NN	O	B-protein
negative	NN	O	I-protein
regulator	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
communication	NN	O	O
,	NN	O	O
we	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
pathway	NN	O	O
of	NN	O	O
induction	NN	O	O
in	NN	O	O
the	NN	O	O
macrophage	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U937	NN	O	B-cell_line
is	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
ICK-1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
does	NN	O	O
not	NN	O	O
confer	NN	O	O
negative	NN	O	O
regulation	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
additional	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
MIP-1	NN	O	B-DNA
alpha	NN	O	I-DNA
nuclear	NN	O	I-DNA
protein	NN	O	I-DNA
(	NN	O	I-DNA
MNP	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
overlaps	NN	O	O
the	NN	O	O
ICK-1	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

Interaction	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
various	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
tissue	NN	O	O
with	NN	O	O
the	NN	O	O
MNP	NN	O	B-DNA
site	NN	O	I-DNA
leads	NN	O	O
to	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
fast-migrating	NN	O	B-protein
protein-DNA	NN	O	I-protein
complexes	NN	O	I-protein
with	NN	O	O
similar	NN	O	O
but	NN	O	O
distinct	NN	O	O
electrophoretic	NN	O	O
mobilities	NN	O	O
.	NN	O	O

A	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
MNP	NN	O	B-DNA
site	NN	O	I-DNA
which	NN	O	O
does	NN	O	O
not	NN	O	O
abrogate	NN	O	O
ICK-1	NN	O	O
binding	NN	O	O
inactivates	NN	O	O
the	NN	O	O
GOS19.1	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
reduces	NN	O	O
its	NN	O	O
activity	NN	O	O
by	NN	O	O
fourfold	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
MNP	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
binding	NN	O	O
at	NN	O	O
the	NN	O	O
MNP	NN	O	B-DNA
site	NN	O	I-DNA
constitutes	NN	O	O
a	NN	O	O
novel	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
expressed	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
Toremifene	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
some	NN	O	O
genes	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Toremifene	NN	O	O
exerts	NN	O	O
multiple	NN	O	O
and	NN	O	O
varied	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

After	NN	O	O
short-term	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
exposure	NN	O	O
to	NN	O	O
therapeutical	NN	O	O
levels	NN	O	O
,	NN	O	O
distinct	NN	O	O
changes	NN	O	O
in	NN	O	O
P-glycoprotein	NN	O	O
,	NN	O	O
steroid	NN	O	O
receptors	NN	O	O
,	NN	O	O
p53	NN	O	O
and	NN	O	O
Bcl-2	NN	O	O
expression	NN	O	O
take	NN	O	O
place	NN	O	O
.	NN	O	O

In	NN	O	O
view	NN	O	O
of	NN	O	O
the	NN	O	O
increasing	NN	O	O
use	NN	O	O
of	NN	O	O
antiestrogens	NN	O	O
in	NN	O	O
cancer	NN	O	O
therapy	NN	O	O
and	NN	O	O
prevention	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
obvious	NN	O	O
merit	NN	O	O
in	NN	O	O
long-term	NN	O	O
in	NN	O	O
vivo	NN	O	O
studies	NN	O	O
to	NN	O	O
be	NN	O	O
conducted	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
interleukin-5	NN	O	B-protein
/receptor	NN	O	O
interaction	NN	O	O
activates	NN	O	O
Lyn	NN	O	B-protein
and	NN	O	I-protein
Jak2	NN	O	I-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
and	NN	O	O
propagates	NN	O	O
signals	NN	O	O
via	NN	O	O
the	NN	O	O
Ras-Raf-1-MAP	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
the	NN	O	O
Jak	NN	O	B-protein
-STAT	NN	O	B-protein
pathways	NN	O	O
in	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-5	NN	O	I-protein
with	NN	O	O
the	NN	O	O
receptor	NN	O	O
activates	NN	O	O
Lyn	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
within	NN	O	O
1	NN	O	O
min	NN	O	O
and	NN	O	O
Jak2	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
within	NN	O	O
1-3	NN	O	O
min	NN	O	O
.	NN	O	O

IL-5	NN	O	B-protein
also	NN	O	O
stimulates	NN	O	O
GTP	NN	O	O
binding	NN	O	O
to	NN	O	O
p21ras	NN	O	B-protein
.	NN	O	O

The	NN	O	O
signal	NN	O	O
is	NN	O	O
subsequently	NN	O	O
propagated	NN	O	O
through	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Raf-1	NN	O	B-protein
,	NN	O	O
MEK	NN	O	B-protein
,	NN	O	O
and	NN	O	O
MAP	NN	O	B-protein
kinases	NN	O	I-protein
as	NN	O	O
shown	NN	O	O
by	NN	O	O
their	NN	O	O
increased	NN	O	O
autophosphorylation	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
situ	NN	O	O
.	NN	O	O

Jak2	NN	O	B-protein
kinase	NN	O	I-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
phosphorylate	NN	O	O
STAT	NN	O	B-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
nuclear	NN	O	I-protein
factors	NN	O	I-protein
was	NN	O	O
studied	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
using	NN	O	O
a	NN	O	O
gamma	NN	O	B-DNA
activation	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
GAS	NN	O	B-DNA
)	NN	O	O
probe	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
IL-5	NN	O	B-protein
induces	NN	O	O
two	NN	O	O
GAS-binding	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
eosinophils	NN	O	B-cell_type
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
STAT1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
IL-5	NN	O	B-protein
induced	NN	O	O
signals	NN	O	O
are	NN	O	O
propagated	NN	O	O
through	NN	O	O
two	NN	O	O
distinct	NN	O	O
pathways	NN	O	O
:	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
Lyn	NN	O	B-protein
--	NN	O	O
>	NN	O	O
Ras	NN	O	B-protein
--	NN	O	O
>	NN	O	O
Raf-1	NN	O	B-protein
--	NN	O	O
>	NN	O	O
MEK	NN	O	B-protein
--	NN	O	O
>	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
Jak2	NN	O	B-protein
--	NN	O	O
>	NN	O	O
STAT1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
retinoblastoma	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
negatively	NN	O	O
regulates	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
human	NN	O	B-protein
cytomegalovirus	NN	O	I-protein
IE2	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
IE2	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
of	NN	O	O
human	NN	O	O
cytomegalovirus	NN	O	O
(	NN	O	O
HCMV	NN	O	O
)	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
a	NN	O	O
few	NN	O	O
viral	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
expressed	NN	O	O
immediately	NN	O	O
upon	NN	O	O
infection	NN	O	O
of	NN	O	O
the	NN	O	O
host	NN	O	B-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
of	NN	O	O
many	NN	O	O
viral	NN	O	B-DNA
and	NN	O	I-DNA
cellular	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-protein
susceptibility	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
(	NN	O	O
Rb	NN	O	B-protein
)	NN	O	O
dramatically	NN	O	O
suppressed	NN	O	O
this	NN	O	O
IE2	NN	O	B-protein
transactivation	NN	O	O
of	NN	O	O
various	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
unlike	NN	O	O
another	NN	O	O
tumor	NN	O	B-protein
suppressor	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
p53	NN	O	B-protein
,	NN	O	O
Rb	NN	O	B-protein
did	NN	O	O
not	NN	O	O
have	NN	O	O
any	NN	O	O
significant	NN	O	O
effect	NN	O	O
on	NN	O	O
basal	NN	O	O
levels	NN	O	O
of	NN	O	O
transcription	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
Rb	NN	O	B-protein
specifically	NN	O	O
interacts	NN	O	O
with	NN	O	O
IE2	NN	O	B-protein
rather	NN	O	O
than	NN	O	O
other	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
general	NN	O	O
transcription	NN	O	O
machinery	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
by	NN	O	O
protein-affinity	NN	O	O
chromatography	NN	O	O
that	NN	O	O
Rb	NN	O	B-protein
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
or	NN	O	O
produced	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
translation	NN	O	O
directly	NN	O	O
bound	NN	O	O
to	NN	O	O
IE2	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
Rb	NN	O	B-protein
may	NN	O	O
regulate	NN	O	O
the	NN	O	O
life	NN	O	O
cycle	NN	O	O
of	NN	O	O
HCMV	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
endemic	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
population	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
may	NN	O	O
provide	NN	O	O
new	NN	O	O
insights	NN	O	O
into	NN	O	O
the	NN	O	O
slow	NN	O	O
rate	NN	O	O
of	NN	O	O
HCMV	NN	O	O
DNA	NN	O	O
replication	NN	O	O
in	NN	O	O
cells	NN	O	O
and	NN	O	O
the	NN	O	O
possible	NN	O	O
involvement	NN	O	O
of	NN	O	O
HCMV	NN	O	O
in	NN	O	O
tumorigenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
nucleoside	NN	O	B-protein
diphosphate	NN	O	I-protein
kinase	NN	O	I-protein
in	NN	O	O
human	NN	O	O
skin	NN	O	O
cancers	NN	O	O
:	NN	O	O
an	NN	O	O
immunohistochemical	NN	O	O
study	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
nucleoside	NN	O	B-protein
diphosphate	NN	O	I-protein
(	NN	O	I-protein
NDP	NN	O	I-protein
)	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
nm23	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
species	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
inversely	NN	O	O
associated	NN	O	O
with	NN	O	O
metastatic	NN	O	O
potential	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
relationship	NN	O	O
remains	NN	O	O
controversial	NN	O	O
according	NN	O	O
to	NN	O	O
the	NN	O	O
tumor	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
and	NN	O	O
experimental	NN	O	O
system	NN	O	O
,	NN	O	O
with	NN	O	O
conflicting	NN	O	O
results	NN	O	O
from	NN	O	O
different	NN	O	O
research	NN	O	O
groups	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
NDP	NN	O	B-protein
kinase	NN	O	I-protein
expression	NN	O	O
serves	NN	O	O
as	NN	O	O
a	NN	O	O
marker	NN	O	O
for	NN	O	O
metastatic	NN	O	O
potential	NN	O	O
in	NN	O	O
human	NN	O	O
skin	NN	O	O
cancer	NN	O	O
,	NN	O	O
we	NN	O	O
assessed	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
NDP	NN	O	B-protein
kinase	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
9	NN	O	O
keratoacanthomas	NN	O	O
(	NN	O	O
KAs	NN	O	O
)	NN	O	O
,	NN	O	O
26	NN	O	O
squamous	NN	O	O
cell	NN	O	O
carcinomas	NN	O	O
(	NN	O	O
SCCs	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
25	NN	O	O
basal	NN	O	O
cell	NN	O	O
carcinomas	NN	O	O
(	NN	O	O
BCCs	NN	O	O
)	NN	O	O
using	NN	O	O
immunohistochemistry	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
NDP	NN	O	B-protein
kinase	NN	O	I-protein
was	NN	O	O
intense	NN	O	O
in	NN	O	O
KA	NN	O	O
and	NN	O	O
SCC	NN	O	O
compared	NN	O	O
with	NN	O	O
BCC	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
difference	NN	O	O
of	NN	O	O
NDP	NN	O	B-protein
kinase	NN	O	I-protein
expression	NN	O	O
between	NN	O	O
KA	NN	O	O
and	NN	O	O
SCC	NN	O	O
was	NN	O	O
not	NN	O	O
statistically	NN	O	O
significant	NN	O	O
.	NN	O	O

And	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
statistically	NN	O	O
significant	NN	O	O
difference	NN	O	O
in	NN	O	O
NDP	NN	O	B-protein
kinase	NN	O	I-protein
expression	NN	O	O
between	NN	O	O
SCC	NN	O	O
with	NN	O	O
metastasis	NN	O	O
and	NN	O	O
SCC	NN	O	O
without	NN	O	O
metastasis	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
contradict	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
concerning	NN	O	O
the	NN	O	O
possible	NN	O	O
role	NN	O	O
of	NN	O	O
nm23	NN	O	B-DNA
gene	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
metastatic	NN	O	O
suppressor	NN	O	O
gene	NN	O	O
in	NN	O	O
human	NN	O	O
skin	NN	O	O
cancer	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
overexpression	NN	O	O
in	NN	O	O
various	NN	O	O
tumor	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
and	NN	O	O
its	NN	O	O
biological	NN	O	O
significance	NN	O	O
in	NN	O	O
cutaneous	NN	O	O
carcinogenesis	NN	O	O
remain	NN	O	O
to	NN	O	O
be	NN	O	O
determined	NN	O	O
.	NN	O	O

-DOCSTART-	O

RB	NN	O	B-protein
and	NN	O	O
a	NN	O	O
novel	NN	O	O
E2F-1	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
MHC	NN	O	B-cell_line
class	NN	O	I-cell_line
II	NN	O	I-cell_line
deficient	NN	O	I-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
normal	NN	O	O
IFN-gamma	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-DNA
IL	NN	O	I-DNA
transactivator	NN	O	I-DNA
CIITA	NN	O	B-DNA
in	NN	O	O
class	NN	O	B-cell_line
II	NN	O	I-cell_line
non-inducible	NN	O	I-cell_line
RB-defective	NN	O	I-cell_line
tumor	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
(	NN	O	I-DNA
MHC	NN	O	I-DNA
)	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
encode	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
proteins	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
antigenic	NN	O	O
peptide	NN	O	O
for	NN	O	O
presentation	NN	O	O
to	NN	O	O
T-cells	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
expressed	NN	O	O
constitutively	NN	O	O
on	NN	O	O
B-cells	NN	O	B-cell_type
and	NN	O	O
EBV-transformed	NN	O	B-cell_line
B-cells	NN	O	I-cell_line
,	NN	O	I-cell_line
and	NN	O	O
are	NN	O	O
inducible	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
on	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	B-cell_type
types	NN	O	I-cell_type
.	NN	O	O

Retinoblastoma	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
RB	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
tumor	NN	O	O
suppressor	NN	O	O
and	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
repressor	NN	O	I-protein
by	NN	O	O
binding	NN	O	O
and	NN	O	O
inactivating	NN	O	O
the	NN	O	O
transactivator	NN	O	B-DNA
E2F-I	NN	O	I-DNA
.	NN	O	O

RB-defective	NN	O	B-cell_line
tumor	NN	O	I-cell_line
lines	NN	O	I-cell_line
are	NN	O	O
non-inducible	NN	O	O
for	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
by	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
or	NN	O	O
very	NN	O	O
weakly	NN	O	O
inducible	NN	O	O
,	NN	O	O
but	NN	O	O
transfection	NN	O	O
of	NN	O	O
2	NN	O	O
different	NN	O	O
lines	NN	O	O
with	NN	O	O
RB	NN	O	B-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
re-establishes	NN	O	O
or	NN	O	O
substantially	NN	O	O
enhances	NN	O	O
class	NN	O	O
II	NN	O	O
inducibility	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
RB	NN	O	B-protein
status	NN	O	O
of	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
B-cell	NN	O	O
mutants	NN	O	O
that	NN	O	O
are	NN	O	O
defective	NN	O	O
in	NN	O	O
class	NN	O	O
II	NN	O	O
expression	NN	O	O
,	NN	O	O
generated	NN	O	O
either	NN	O	O
in	NN	O	O
vitro	NN	O	O
or	NN	O	O
derived	NN	O	O
from	NN	O	O
Bare	NN	O	O
Lymphocyte	NN	O	O
Syndrome	NN	O	O
(	NN	O	O
BLS	NN	O	O
)	NN	O	O
patients	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
matrix-bound	NN	O	O
RB	NN	O	B-protein
was	NN	O	O
detectable	NN	O	O
in	NN	O	O
all	NN	O	O
cases	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
loss	NN	O	O
of	NN	O	O
RB	NN	O	B-protein
is	NN	O	O
not	NN	O	O
responsible	NN	O	O
for	NN	O	O
decreased	NN	O	O
class	NN	O	O
II	NN	O	O
expression	NN	O	O
in	NN	O	O
these	NN	O	O
lines	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
E2F-I	NN	O	O
binding	NN	O	O
protein	NN	O	O
,	NN	O	O
most	NN	O	O
likely	NN	O	O
DP-I	NN	O	O
,	NN	O	O
was	NN	O	O
also	NN	O	O
apparently	NN	O	O
normal	NN	O	O
in	NN	O	O
both	NN	O	O
class	NN	O	O
II-positive	NN	O	O
and	NN	O	O
-negative	NN	O	O
B-cell	NN	O	O
lines	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
examined	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
CIITA	NN	O	B-DNA
in	NN	O	O
RB	NN	O	B-protein
-defective	NN	O	O
lines	NN	O	O
.	NN	O	O

CIITA	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
transactivator	NN	O	I-DNA
known	NN	O	O
to	NN	O	O
be	NN	O	O
defective	NN	O	O
in	NN	O	O
one	NN	O	O
form	NN	O	O
of	NN	O	O
BLS	NN	O	O
and	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
by	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

CIITA	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
normally	NN	O	O
inducible	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
class	NN	O	B-cell_line
II	NN	O	I-cell_line
non-inducible	NN	O	I-cell_line
,	NN	O	I-cell_line
RB-defective	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
in	NN	O	O
one	NN	O	O
line	NN	O	O
,	NN	O	O
re-expression	NN	O	O
of	NN	O	O
RB	NN	O	B-protein
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
CIITA	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
levels	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
block	NN	O	O
in	NN	O	O
MHC	NN	O	O
class	NN	O	O
II	NN	O	O
inducibility	NN	O	O
in	NN	O	O
RB-defective	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
block	NN	O	O
in	NN	O	O
CIITA	NN	O	B-DNA
inducibility	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin	NN	O	B-protein
12	NN	O	I-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT4	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Interleukin	NN	O	B-protein
12	NN	O	I-protein
(	NN	O	O
IL-12	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
immunoregulatory	NN	O	B-protein
cytokine	NN	O	I-protein
whose	NN	O	O
receptor	NN	O	O
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
hematopoietin	NN	O	B-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
IL-12	NN	O	B-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
Janus	NN	O	B-protein
family	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
JAK2	NN	O	B-protein
and	NN	O	O
Tyk2	NN	O	B-protein
,	NN	O	O
implicating	NN	O	O
these	NN	O	O
kinases	NN	O	B-protein
in	NN	O	O
the	NN	O	O
immediate	NN	O	O
biochemical	NN	O	O
response	NN	O	O
to	NN	O	O
IL-12	NN	O	B-protein
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
known	NN	O	O
as	NN	O	O
STATs	NN	O	B-protein
(	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
)	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
bind	NN	O	O
hematopoietin	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
activate	NN	O	O
JAK	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
IL-12	NN	O	B-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
a	NN	O	O
recently	NN	O	O
identified	NN	O	O
STAT	NN	O	B-protein
family	NN	O	I-protein
member	NN	O	I-protein
,	NN	O	O
STAT4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
show	NN	O	O
that	NN	O	O
STAT4	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
IL-12	NN	O	B-protein
stimulates	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
complex	NN	O	I-protein
that	NN	O	O
recognizes	NN	O	O
a	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
previously	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
that	NN	O	O
this	NN	O	O
complex	NN	O	O
contains	NN	O	O
STAT4	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
recent	NN	O	O
demonstration	NN	O	O
of	NN	O	O
JAK	NN	O	B-protein
phosphorylation	NN	O	O
by	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
identify	NN	O	O
a	NN	O	O
rapid	NN	O	O
signal-transduction	NN	O	O
pathway	NN	O	O
likely	NN	O	O
to	NN	O	O
mediate	NN	O	O
IL-12	NN	O	B-protein
-induced	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Temperature-induced	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
leucocyte	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
sepsis	NN	O	O
or	NN	O	O
septic	NN	O	O
shock	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
Activation	NN	O	O
of	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
axis	NN	O	O
is	NN	O	O
of	NN	O	O
vital	NN	O	O
importance	NN	O	O
during	NN	O	O
critical	NN	O	O
illness	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
adaptive	NN	O	O
mechanisms	NN	O	O
which	NN	O	O
occur	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
glucocorticoid	NN	O	O
target	NN	O	O
tissues	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
sepsis	NN	O	O
or	NN	O	O
septic	NN	O	O
shock	NN	O	O
.	NN	O	O

DESIGN	NN	O	O
:	NN	O	O
The	NN	O	O
effects	NN	O	O
of	NN	O	O
hypercortisolaemia	NN	O	O
,	NN	O	O
hyperthermia	NN	O	O
and	NN	O	O
cellular	NN	O	O
composition	NN	O	O
on	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
per	NN	O	O
cell	NN	O	O
and	NN	O	O
their	NN	O	O
affinity	NN	O	O
were	NN	O	O
evaluated	NN	O	O
,	NN	O	O
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
of	NN	O	O
control	NN	O	O
subjects	NN	O	O
and	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
sepsis	NN	O	O
or	NN	O	O
septic	NN	O	O
shock	NN	O	O
.	NN	O	O

SUBJECTS	NN	O	O
:	NN	O	O
Fifteen	NN	O	O
patients	NN	O	O
(	NN	O	O
age	NN	O	O
25-79	NN	O	O
)	NN	O	O
with	NN	O	O
sepsis	NN	O	O
or	NN	O	O
septic	NN	O	O
shock	NN	O	O
who	NN	O	O
were	NN	O	O
admitted	NN	O	O
to	NN	O	O
an	NN	O	O
intensive	NN	O	O
care	NN	O	O
unit	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

The	NN	O	O
control	NN	O	O
group	NN	O	O
consisted	NN	O	O
of	NN	O	O
24	NN	O	O
healthy	NN	O	O
laboratory	NN	O	O
employees	NN	O	O
.	NN	O	O

MEASUREMENTS	NN	O	O
:	NN	O	O
The	NN	O	O
binding	NN	O	O
capacity	NN	O	O
and	NN	O	O
affinity	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
were	NN	O	O
measured	NN	O	O
and	NN	O	O
compared	NN	O	O
to	NN	O	O
clinical	NN	O	O
data	NN	O	O
and	NN	O	O
the	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Hypercortisolaemia	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
decreased	NN	O	O
affinity	NN	O	O
and	NN	O	O
a	NN	O	O
decreased	NN	O	O
binding	NN	O	O
capacity	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
,	NN	O	O
hyperthermia	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
variations	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
composition	NN	O	O
did	NN	O	O
not	NN	O	O
influence	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
receptors	NN	O	O
per	NN	O	O
cell	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
sepsis	NN	O	O
or	NN	O	O
septic	NN	O	O
shock	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
decreased	NN	O	O
affinity	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
was	NN	O	O
observed	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
weak	NN	O	O
but	NN	O	O
significant	NN	O	O
negative	NN	O	O
correlation	NN	O	O
between	NN	O	O
body	NN	O	O
temperature	NN	O	O
and	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
patient	NN	O	O
group	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
relation	NN	O	O
between	NN	O	O
circulating	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
affinity	NN	O	O
and	NN	O	O
number	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
There	NN	O	O
is	NN	O	O
no	NN	O	O
obvious	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
by	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

The	NN	O	O
decreased	NN	O	O
affinity	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
together	NN	O	O
with	NN	O	O
the	NN	O	O
negative	NN	O	O
correlation	NN	O	O
between	NN	O	O
hyperthermia	NN	O	O
and	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
patients	NN	O	O
with	NN	O	O
sepsis	NN	O	O
or	NN	O	O
septic	NN	O	O
shock	NN	O	O
suggest	NN	O	O
that	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
axis	NN	O	O
activation	NN	O	O
during	NN	O	O
critical	NN	O	O
illness	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
peripheral	NN	O	O
adaptation	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
number	NN	O	O
and	NN	O	O
affinity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Integrin-mediated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
message	NN	O	O
induction	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
possible	NN	O	O
signaling	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
Syk	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
aspect	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
mediated	NN	O	O
by	NN	O	O
integrins	NN	O	B-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
,	NN	O	O
either	NN	O	O
integrin	NN	O	B-protein
-dependent	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
to	NN	O	O
fibronectin	NN	O	B-protein
or	NN	O	O
ligation	NN	O	O
of	NN	O	O
beta	NN	O	O
1	NN	O	O
integrins	NN	O	O
with	NN	O	O
antibodies	NN	O	B-protein
causes	NN	O	O
a	NN	O	O
rapid	NN	O	O
and	NN	O	O
intense	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
two	NN	O	O
sets	NN	O	O
of	NN	O	O
proteins	NN	O	O
of	NN	O	O
about	NN	O	O
65-75	NN	O	B-protein
and	NN	O	I-protein
120-125	NN	O	I-protein
kDa	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
integrin	NN	O	B-protein
ligation	NN	O	O
leads	NN	O	O
to	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
subunits	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
driven	NN	O	O
by	NN	O	O
a	NN	O	O
promoter	NN	O	B-DNA
containing	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
to	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
mRNAs	NN	O	B-RNA
for	NN	O	O
immediate-early	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
interleukin	NN	O	I-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
beta	NN	O	I-protein
.	NN	O	O

The	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
inhibitors	NN	O	O
genistein	NN	O	O
and	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
block	NN	O	O
both	NN	O	O
integrin	NN	O	B-protein
-mediated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
increases	NN	O	O
in	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
message	NN	O	O
levels	NN	O	O
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
causal	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
events	NN	O	O
.	NN	O	O

The	NN	O	O
components	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
subsequent	NN	O	O
to	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
include	NN	O	O
paxillin	NN	O	B-protein
,	NN	O	O
pp125FAK	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
domain	NN	O	I-protein
containing	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
Syk	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
integrin	NN	O	B-protein
ligation	NN	O	O
with	NN	O	O
antibodies	NN	O	B-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Syk	NN	O	B-protein
but	NN	O	O
not	NN	O	O
of	NN	O	O
FAK	NN	O	B-protein
or	NN	O	O
paxillin	NN	O	B-protein
.	NN	O	O

In	NN	O	O
adhering	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
pre-treatment	NN	O	O
with	NN	O	O
cytochalasin	NN	O	O
D	NN	O	O
suppresses	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
FAK	NN	O	B-protein
and	NN	O	O
paxillin	NN	O	B-protein
but	NN	O	O
not	NN	O	O
of	NN	O	O
Syk	NN	O	B-protein
,	NN	O	O
while	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
message	NN	O	O
induction	NN	O	O
is	NN	O	O
unaffected	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
Syk	NN	O	B-protein
tyrosine	NN	O	O
kinase	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
component	NN	O	O
of	NN	O	O
an	NN	O	O
integrin	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
leading	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
to	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
messages	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
to	NN	O	O
human	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
by	NN	O	O
New	NN	O	O
World	NN	O	O
primate	NN	O	O
cell	NN	O	O
extracts	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
if	NN	O	O
New	NN	O	O
World	NN	O	O
primates	NN	O	O
express	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
that	NN	O	O
influences	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
in	NN	O	O
cytosol	NN	O	O
prepared	NN	O	O
from	NN	O	O
B95-8	NN	O	B-cell_line
lymphoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
cotton	NN	O	O
top	NN	O	O
tamarin	NN	O	O
(	NN	O	O
Saguinus	NN	O	O
oedipus	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
cytosol	NN	O	O
prepared	NN	O	O
from	NN	O	O
human	NN	O	O
or	NN	O	O
rat	NN	O	O
tissues	NN	O	O
.	NN	O	O

B95-8	NN	O	O
cytosol	NN	O	O
inhibited	NN	O	O
specific	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
when	NN	O	O
mixed	NN	O	O
with	NN	O	O
cytosol	NN	O	O
prepared	NN	O	O
from	NN	O	O
either	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
lymphoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
HL	NN	O	B-cell_line
)	NN	O	O
or	NN	O	O
rat	NN	O	O
thymus	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
activity	NN	O	O
was	NN	O	O
heat	NN	O	O
labile	NN	O	O
and	NN	O	O
trypsin	NN	O	B-protein
sensitive	NN	O	O
.	NN	O	O

Peak	NN	O	O
inhibitory	NN	O	O
activity	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
150-200	NN	O	O
kd	NN	O	O
fractions	NN	O	O
after	NN	O	O
Sephacryl	NN	O	O
G-200	NN	O	O
ultrafiltration	NN	O	O
.	NN	O	O

Scatchard	NN	O	O
analysis	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
using	NN	O	O
mixed	NN	O	O
cytosol	NN	O	O
showed	NN	O	O
a	NN	O	O
diminished	NN	O	O
GR	NN	O	B-protein
apparent	NN	O	O
binding	NN	O	O
affinity	NN	O	O
when	NN	O	O
compared	NN	O	O
to	NN	O	O
HL	NN	O	O
cytosol	NN	O	O
.	NN	O	O

Kinetic	NN	O	O
studies	NN	O	O
using	NN	O	O
mixed	NN	O	O
cytosol	NN	O	O
indicated	NN	O	O
that	NN	O	O
B95-8	NN	O	O
cytosol	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
apparent	NN	O	O
dissociation	NN	O	O
rate	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
B95-8	NN	O	B-cell_type
cells	NN	O	I-cell_type
contain	NN	O	O
a	NN	O	O
competitive	NN	O	O
inhibitor	NN	O	O
that	NN	O	O
prevents	NN	O	O
binding	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
to	NN	O	O
its	NN	O	O
cognate	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Disruption	NN	O	O
of	NN	O	O
a	NN	O	O
GATA	NN	O	B-DNA
motif	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
Duffy	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
abolishes	NN	O	O
erythroid	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
Duffy-negative	NN	O	O
individuals	NN	O	O
.	NN	O	O

The	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
the	NN	O	O
Duffy	NN	O	B-protein
blood	NN	O	I-protein
group	NN	O	I-protein
antigen	NN	O	I-protein
,	NN	O	O
the	NN	O	O
erythrocyte	NN	O	B-protein
receptor	NN	O	I-protein
for	NN	O	O
the	NN	O	O
Plasmodium	NN	O	O
vivax	NN	O	O
malaria	NN	O	O
parasite	NN	O	O
,	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
cloned	NN	O	O
and	NN	O	O
shown	NN	O	O
to	NN	O	O
encode	NN	O	O
a	NN	O	O
widely	NN	O	O
expressed	NN	O	O
chemokine	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
Duffy	NN	O	B-DNA
antigen/chemokine	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
DARC	NN	O	B-DNA
)	NN	O	O
is	NN	O	O
composed	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
exon	NN	O	B-DNA
and	NN	O	O
that	NN	O	O
most	NN	O	O
Duffy-negative	NN	O	O
blacks	NN	O	O
carry	NN	O	O
a	NN	O	O
silent	NN	O	B-DNA
FY*B	NN	O	I-DNA
allele	NN	O	I-DNA
with	NN	O	O
a	NN	O	O
single	NN	O	O
T	NN	O	O
to	NN	O	O
C	NN	O	O
substitution	NN	O	O
at	NN	O	O
nucleotide	NN	O	O
-46	NN	O	O
.	NN	O	O

This	NN	O	O
mutation	NN	O	O
impairs	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
disrupting	NN	O	O
a	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
GATA1	NN	O	B-protein
erythroid	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

With	NN	O	O
the	NN	O	O
recent	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
FY*A	NN	O	B-DNA
and	NN	O	I-DNA
FY*B	NN	O	I-DNA
alleles	NN	O	I-DNA
,	NN	O	O
these	NN	O	O
findings	NN	O	O
provide	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
Duffy	NN	O	O
blood	NN	O	O
group	NN	O	O
system	NN	O	O
and	NN	O	O
an	NN	O	O
explanation	NN	O	O
for	NN	O	O
the	NN	O	O
erythroid-specific	NN	O	O
repression	NN	O	O
of	NN	O	O
the	NN	O	O
DARC	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
Duffy-negative	NN	O	O
individuals	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
pp90rsk	NN	O	B-protein
and	NN	O	O
early	NN	O	B-DNA
growth	NN	O	I-DNA
response-1	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
pokeweed	NN	O	B-protein
mitogen	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
present	NN	O	O
studies	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
pokeweed	NN	O	B-protein
mitogen	NN	O	I-protein
(	NN	O	O
PWM	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
early	NN	O	B-DNA
growth	NN	O	I-DNA
response-1	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
EGR-1	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

PWM	NN	O	B-protein
regulates	NN	O	O
EGR-1	NN	O	B-DNA
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
both	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
post-transcriptional	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
with	NN	O	O
EGR-1	NN	O	B-DNA
promoter	NN	O	I-DNA
fragments	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
the	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
PWM	NN	O	B-protein
induced	NN	O	O
EGR-1	NN	O	B-DNA
transcription	NN	O	O
is	NN	O	O
conferred	NN	O	O
by	NN	O	O
the	NN	O	O
CArG	NN	O	B-DNA
motif	NN	O	I-DNA
(	NN	O	O
C	NN	O	B-DNA
C	NN	O	I-DNA
[	NN	O	I-DNA
AT	NN	O	I-DNA
]	NN	O	I-DNA
6GG	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
EGR-1	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
results	NN	O	O
further	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
S6	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
pp90rsk	NN	O	B-protein
)	NN	O	O
,	NN	O	O
evidenced	NN	O	O
by	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
S6	NN	O	B-protein
and	NN	O	O
serum	NN	O	O
response	NN	O	O
factor	NN	O	O
(	NN	O	O
SRF	NN	O	O
)	NN	O	O
peptides	NN	O	O
,	NN	O	O
in	NN	O	O
PWM	NN	O	B-cell_line
treated	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
PWM	NN	O	B-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
initiate	NN	O	O
an	NN	O	O
intracytoplasmic	NN	O	O
signalling	NN	O	O
cascade	NN	O	O
and	NN	O	O
EGR-1	NN	O	B-DNA
induction	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
conserved	NN	O	B-DNA
motif	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
several	NN	O	O
cytokines	NN	O	B-protein
expressed	NN	O	O
by	NN	O	O
human	NN	O	B-cell_line
Th2-type	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
found	NN	O	O
a	NN	O	O
novel	NN	O	O
conserved	NN	O	B-DNA
motif	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
several	NN	O	O
T-cell-expressed	NN	O	B-protein
cytokines	NN	O	I-protein
[	NN	O	O
human	NN	O	B-protein
interleukin-2	NN	O	I-protein
,	NN	O	I-protein
-4	NN	O	I-protein
,	NN	O	I-protein
-5	NN	O	I-protein
and	NN	O	I-protein
-13	NN	O	I-protein
and	NN	O	O
human	NN	O	B-protein
and	NN	O	I-protein
mouse	NN	O	I-protein
granulocyte/macrophage-colony	NN	O	I-protein
stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
]	NN	O	O
.	NN	O	O

It	NN	O	O
contains	NN	O	O
a	NN	O	O
core	NN	O	B-DNA
sequence	NN	O	I-DNA
CTTGG	NN	O	O
...	NN	O	O
CCAAG	NN	O	O
which	NN	O	O
is	NN	O	O
present	NN	O	O
as	NN	O	O
part	NN	O	O
of	NN	O	O
larger	NN	O	O
palindromic	NN	O	B-DNA
sequences	NN	O	I-DNA
in	NN	O	O
each	NN	O	O
gene	NN	O	O
.	NN	O	O

This	NN	O	O
suggest	NN	O	O
that	NN	O	O
they	NN	O	O
may	NN	O	O
interact	NN	O	O
with	NN	O	O
a	NN	O	O
new	NN	O	O
family	NN	O	O
of	NN	O	O
trans-acting	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
transfection	NN	O	O
assays	NN	O	O
,	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
element	NN	O	I-DNA
has	NN	O	O
a	NN	O	O
strong	NN	O	O
positive	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Jurkat	NN	O	B-cell_line
J6	NN	O	I-cell_line
upon	NN	O	O
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
DNA	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
this	NN	O	O
sequence	NN	O	O
can	NN	O	O
give	NN	O	O
either	NN	O	O
six	NN	O	O
different	NN	O	O
specific	NN	O	O
bands	NN	O	O
which	NN	O	O
are	NN	O	O
competed	NN	O	O
out	NN	O	O
by	NN	O	O
different	NN	O	O
parts	NN	O	O
of	NN	O	O
the	NN	O	O
sequence	NN	O	O
or	NN	O	O
one	NN	O	O
specific	NN	O	O
band	NN	O	O
which	NN	O	O
is	NN	O	O
competed	NN	O	O
out	NN	O	O
by	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
inverted	NN	O	O
repeats	NN	O	O
,	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
reconstitution	NN	O	O
conditions	NN	O	O
.	NN	O	O

In	NN	O	O
different	NN	O	O
genes	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
sequences	NN	O	I-DNA
are	NN	O	O
separated	NN	O	O
by	NN	O	O
integer	NN	O	O
numbers	NN	O	O
of	NN	O	O
helical	NN	O	O
turns	NN	O	O
.	NN	O	O

Considering	NN	O	O
the	NN	O	O
strong	NN	O	O
positive	NN	O	O
regulatory	NN	O	O
effect	NN	O	O
of	NN	O	O
this	NN	O	O
element	NN	O	B-DNA
and	NN	O	O
its	NN	O	O
presence	NN	O	O
in	NN	O	O
several	NN	O	O
T-cell-expressed	NN	O	B-DNA
cytokine	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
it	NN	O	O
may	NN	O	O
be	NN	O	O
crucial	NN	O	O
to	NN	O	O
the	NN	O	O
coordinated	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
helper	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

cDNA	NN	O	B-DNA
cloning	NN	O	O
of	NN	O	O
a	NN	O	O
NGFI-B/nur77-related	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
from	NN	O	O
an	NN	O	O
apoptotic	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
PEER	NN	O	B-cell_line
,	NN	O	O
dies	NN	O	O
by	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
PMA	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
gene	NN	O	B-DNA
,	NN	O	O
TINUR	NN	O	B-DNA
,	NN	O	O
was	NN	O	O
cloned	NN	O	O
from	NN	O	O
apoptotic	NN	O	B-cell_line
PEER	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
TINUR	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
induced	NN	O	O
within	NN	O	O
1	NN	O	O
h	NN	O	O
after	NN	O	O
the	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
Ag	NN	O	I-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

TINUR	NN	O	B-DNA
belongs	NN	O	O
to	NN	O	O
the	NN	O	O
NGFI-B/nur77	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
the	NN	O	O
steroid	NN	O	B-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
and	NN	O	O
is	NN	O	O
an	NN	O	O
orphan	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

TINUR	NN	O	B-DNA
binds	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
DNA	NN	O	B-DNA
sequence	NN	O	I-DNA
as	NN	O	O
NGFI-B/nur77	NN	O	B-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
NGFI-B/nur77	NN	O	B-protein
family	NN	O	I-protein
can	NN	O	O
be	NN	O	O
classified	NN	O	O
into	NN	O	O
two	NN	O	O
subtypes	NN	O	O
.	NN	O	O

-DOCSTART-	O

Aspirin	NN	O	O
inhibits	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
mobilization	NN	O	O
and	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
in	NN	O	O
stimulated	NN	O	B-cell_line
human	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
The	NN	O	O
induction	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
E-selectin	NN	O	B-protein
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
mobilization	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Since	NN	O	O
salicylates	NN	O	O
have	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
inhibit	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
preventing	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
its	NN	O	O
inhibitor	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
we	NN	O	O
studied	NN	O	O
a	NN	O	O
potential	NN	O	O
inhibition	NN	O	O
of	NN	O	O
this	NN	O	O
pathway	NN	O	O
by	NN	O	O
acetylsalicylate	NN	O	O
(	NN	O	O
aspirin	NN	O	O
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
umbilical	NN	O	I-cell_line
vein	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
HUVECs	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

METHODS	NN	O	O
AND	NN	O	O
RESULTS	NN	O	O
:	NN	O	O
Gel-shift	NN	O	O
analyses	NN	O	O
demonstrated	NN	O	O
dose-dependent	NN	O	O
inhibition	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
mobilization	NN	O	O
by	NN	O	O
aspirin	NN	O	O
at	NN	O	O
concentrations	NN	O	O
ranging	NN	O	O
from	NN	O	O
1	NN	O	O
to	NN	O	O
10	NN	O	O
mmol/L	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
and	NN	O	O
E-selectin	NN	O	B-protein
surface	NN	O	O
expression	NN	O	O
by	NN	O	O
TNF	NN	O	B-protein
was	NN	O	O
dose-dependently	NN	O	O
reduced	NN	O	O
by	NN	O	O
aspirin	NN	O	O
over	NN	O	O
the	NN	O	O
same	NN	O	O
range	NN	O	O
,	NN	O	O
while	NN	O	O
induction	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
was	NN	O	O
hardly	NN	O	O
affected	NN	O	O
.	NN	O	O

Aspirin	NN	O	O
appeared	NN	O	O
to	NN	O	O
prevent	NN	O	O
VCAM-1	NN	O	B-protein
transcription	NN	O	O
,	NN	O	O
since	NN	O	O
it	NN	O	O
dose-dependently	NN	O	O
inhibited	NN	O	O
induction	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
TNF	NN	O	B-protein
.	NN	O	O

As	NN	O	O
a	NN	O	O
functional	NN	O	O
consequence	NN	O	O
,	NN	O	O
adhesion	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
to	NN	O	O
TNF-stimulated	NN	O	B-cell_line
HUVECs	NN	O	I-cell_line
was	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
by	NN	O	O
aspirin	NN	O	O
due	NN	O	O
to	NN	O	O
suppression	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
and	NN	O	O
E-selectin	NN	O	B-protein
upregulation	NN	O	O
.	NN	O	O

These	NN	O	O
effects	NN	O	O
of	NN	O	O
aspirin	NN	O	O
were	NN	O	O
not	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
cyclooxygenase	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
since	NN	O	O
indomethacin	NN	O	O
was	NN	O	O
ineffective	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
aspirin	NN	O	O
inhibits	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
mobilization	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
and	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
subsequent	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
thereby	NN	O	O
providing	NN	O	O
an	NN	O	O
additional	NN	O	O
mechanism	NN	O	O
for	NN	O	O
therapeutic	NN	O	O
effects	NN	O	O
of	NN	O	O
aspirin	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
phosphatase	NN	O	B-protein
inhibitors	NN	O	O
involves	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
at	NN	O	O
phosphatase	NN	O	B-protein
2A-sensitive	NN	O	I-protein
sites	NN	O	I-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
various	NN	O	O
cellular	NN	O	O
stimuli	NN	O	O
involves	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
subsequent	NN	O	O
degradation	NN	O	O
of	NN	O	O
its	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
although	NN	O	O
the	NN	O	O
underlying	NN	O	O
mechanism	NN	O	O
remains	NN	O	O
unclear	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
serine/threonine	NN	O	B-protein
phosphatases	NN	O	I-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
phosphorylation	NN	O	O
was	NN	O	O
investigated	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
incubation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
low	NN	O	O
concentrations	NN	O	O
(	NN	O	O
approximately	NN	O	O
1-5	NN	O	O
nM	NN	O	O
)	NN	O	O
of	NN	O	O
calyculin	NN	O	O
A	NN	O	O
or	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
,	NN	O	O
potent	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
phosphatase	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
PP-1	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
type	NN	O	I-protein
2A	NN	O	I-protein
(	NN	O	O
PP-2A	NN	O	B-protein
)	NN	O	O
,	NN	O	O
induces	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
even	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
any	NN	O	O
cellular	NN	O	O
stimulus	NN	O	O
.	NN	O	O

This	NN	O	O
action	NN	O	O
of	NN	O	O
the	NN	O	O
phosphatase	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
RelA.p50	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
heterodimer	NN	O	I-protein
,	NN	O	O
is	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
agents	NN	O	O
that	NN	O	O
block	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
phosphorylation	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
phosphorylated	NN	O	B-protein
I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
from	NN	O	O
calyculin	NN	O	B-cell_line
A-treated	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
that	NN	O	O
from	NN	O	O
TNF-alpha-stimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
is	NN	O	O
sensitive	NN	O	O
to	NN	O	O
PP-2A	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
fundamental	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
the	NN	O	O
two	NN	O	O
different	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inducers	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
phosphorylation	NN	O	O
by	NN	O	O
both	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
the	NN	O	O
phosphatase	NN	O	O
inhibitors	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
subsequent	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
TNF-alpha-	NN	O	O
and	NN	O	O
calyculin	NN	O	O
A-induced	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
alpha	NN	O	O
degradation	NN	O	O
exhibits	NN	O	O
similar	NN	O	O
but	NN	O	O
not	NN	O	O
identical	NN	O	O
sensitivities	NN	O	O
to	NN	O	O
a	NN	O	O
proteasome	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
mediated	NN	O	O
through	NN	O	O
both	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-protein
-inducible	NN	O	O
and	NN	O	O
the	NN	O	O
PP-2A-opposing	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
may	NN	O	O
serve	NN	O	O
to	NN	O	O
target	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
for	NN	O	O
proteasome	NN	O	B-protein
-mediated	NN	O	O
degradation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Sp1	NN	O	B-protein
functions	NN	O	O
in	NN	O	O
a	NN	O	O
chromatin	NN	O	B-DNA
-dependent	NN	O	O
manner	NN	O	O
to	NN	O	O
augment	NN	O	O
human	NN	O	B-DNA
alpha-globin	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

The	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
alpha-globin	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
highly	NN	O	O
G	NN	O	O
+	NN	O	O
C	NN	O	O
rich	NN	O	O
and	NN	O	O
contains	NN	O	O
multiple	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
sequence	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
Sp1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
G	NN	O	B-DNA
+	NN	O	I-DNA
C-rich	NN	O	I-DNA
region	NN	O	I-DNA
in	NN	O	O
augmenting	NN	O	O
alpha-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
far-upstream	NN	O	B-DNA
alpha-globin	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
HS-40	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
transiently	NN	O	B-cell_line
transfected	NN	O	I-cell_line
erythroid	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
alpha-globin	NN	O	B-DNA
G	NN	O	I-DNA
+	NN	O	I-DNA
C-rich	NN	O	I-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
region	NN	O	I-DNA
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
alpha-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
upon	NN	O	O
stable	NN	O	O
integration	NN	O	O
into	NN	O	O
chromatin	NN	O	B-DNA
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
causes	NN	O	O
a	NN	O	O
nearly	NN	O	O
90	NN	O	O
%	NN	O	O
decrease	NN	O	O
in	NN	O	O
promoter	NN	O	O
activity	NN	O	O
compared	NN	O	O
with	NN	O	O
expression	NN	O	O
from	NN	O	O
an	NN	O	O
alpha-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
retaining	NN	O	O
this	NN	O	O
region	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
alpha-globin	NN	O	B-DNA
G	NN	O	I-DNA
+	NN	O	I-DNA
C-rich	NN	O	I-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
region	NN	O	I-DNA
augments	NN	O	O
alpha-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
a	NN	O	O
chromatin	NN	O	B-DNA
-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
G	NN	O	B-DNA
+	NN	O	I-DNA
C-rich	NN	O	I-DNA
region	NN	O	I-DNA
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
alpha-globin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
during	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
by	NN	O	O
both	NN	O	O
footprint	NN	O	O
analysis	NN	O	O
and	NN	O	O
functional	NN	O	O
assays	NN	O	O
that	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
G	NN	O	B-DNA
+	NN	O	I-DNA
C-rich	NN	O	I-DNA
region	NN	O	I-DNA
to	NN	O	O
increase	NN	O	O
alpha-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
from	NN	O	O
a	NN	O	O
stably	NN	O	O
integrated	NN	O	O
alpha-globin	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
its	NN	O	O
multiple	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Sp1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Costimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
LFA-3	NN	O	B-protein
and	NN	O	O
B7	NN	O	B-protein
induce	NN	O	O
distinct	NN	O	O
effects	NN	O	O
on	NN	O	O
AP-1	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
earlier	NN	O	O
shown	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
SEA	NN	O	O
presented	NN	O	O
on	NN	O	O
Chinese	NN	O	B-cell_line
hamster	NN	O	I-cell_line
ovary	NN	O	I-cell_line
(	NN	O	I-cell_line
CHO	NN	O	I-cell_line
)	NN	O	I-cell_line
-DR	NN	O	I-cell_line
transfectants	NN	O	I-cell_line
coexpressing	NN	O	O
either	NN	O	O
B7	NN	O	B-protein
or	NN	O	O
LFA-3	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
distinct	NN	O	O
cytokine	NN	O	B-protein
profiles	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
B7	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
LFA-3	NN	O	B-protein
,	NN	O	O
strongly	NN	O	O
costimulated	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
and	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Maximal	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
was	NN	O	O
recorded	NN	O	O
with	NN	O	O
CHO-DR/B7/LFA-3	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
cooperative	NN	O	O
effect	NN	O	O
of	NN	O	O
B7	NN	O	B-protein
and	NN	O	O
LFA-3	NN	O	B-protein
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

Gel-shift	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
CHO-DR	NN	O	B-protein
and	NN	O	O
staphylococcal	NN	O	O
enterotoxin	NN	O	O
A	NN	O	O
was	NN	O	O
sufficient	NN	O	O
to	NN	O	O
induce	NN	O	O
significant	NN	O	O
amounts	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
proteins	NN	O	O
,	NN	O	O
whereas	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
required	NN	O	O
costimulation	NN	O	O
.	NN	O	O

LFA-3	NN	O	B-protein
induced	NN	O	O
moderate	NN	O	O
levels	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
influence	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
while	NN	O	O
B7	NN	O	B-protein
costimulation	NN	O	O
strongly	NN	O	O
induced	NN	O	O
both	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
substantially	NN	O	O
enhanced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
CHO-DR/B7/LFA-3	NN	O	B-cell_line
triple	NN	O	I-cell_line
transfectant	NN	O	I-cell_line
induced	NN	O	O
a	NN	O	O
further	NN	O	O
increase	NN	O	O
in	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	B-protein
proteins	NN	O	I-protein
compared	NN	O	O
with	NN	O	O
the	NN	O	O
double	NN	O	B-cell_line
transfectants	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
Oct-1	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
remained	NN	O	O
similar	NN	O	O
in	NN	O	O
all	NN	O	O
samples	NN	O	O
.	NN	O	O

Super-shift	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
of	NN	O	O
costimulated	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
contained	NN	O	O
large	NN	O	O
amounts	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
substantial	NN	O	O
amounts	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
marginal	NN	O	O
levels	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
contained	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
Jun-D	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Fra-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
marginal	NN	O	O
amounts	NN	O	O
of	NN	O	O
Jun-B	NN	O	B-protein
and	NN	O	O
c-Fos	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
distinct	NN	O	O
effects	NN	O	O
of	NN	O	O
B7	NN	O	B-protein
and	NN	O	O
LFA-3	NN	O	B-protein
costimulation	NN	O	O
on	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

These	NN	O	O
may	NN	O	O
partly	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
differential	NN	O	O
effects	NN	O	O
of	NN	O	O
B7	NN	O	B-protein
and	NN	O	O
LFA-3	NN	O	B-protein
costimulation	NN	O	O
on	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Ah	NN	O	B-protein
receptor	NN	O	I-protein
recognizes	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
BSAP	NN	O	B-protein
:	NN	O	O
a	NN	O	O
possible	NN	O	O
mechanism	NN	O	O
for	NN	O	O
dioxin-mediated	NN	O	O
alteration	NN	O	O
of	NN	O	O
CD19	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8-tetrachlorodibenzo-p-dioxin	NN	O	O
(	NN	O	O
TCDD	NN	O	O
)	NN	O	O
inhibits	NN	O	O
murine	NN	O	O
and	NN	O	O
human	NN	O	O
B	NN	O	O
lymphocyte	NN	O	O
immunoglobulin	NN	O	O
production	NN	O	O
through	NN	O	O
an	NN	O	O
unknown	NN	O	O
mechanism	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
TCDD	NN	O	O
on	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CD19	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
TCDD	NN	O	O
treatment	NN	O	O
decreased	NN	O	O
steady	NN	O	O
state	NN	O	O
levels	NN	O	O
of	NN	O	O
CD19	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
67	NN	O	O
%	NN	O	O
in	NN	O	O
the	NN	O	O
IM-9	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Using	NN	O	O
a	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
complex	NN	O	I-protein
in	NN	O	O
IM-9	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
that	NN	O	O
by	NN	O	O
several	NN	O	O
criteria	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
Ah	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
Ah	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
recognized	NN	O	O
a	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
for	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
lineage-specific	NN	O	I-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
BSAP	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	O
CD19	NN	O	B-DNA
gene	NN	O	I-DNA
which	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
Ah	NN	O	I-DNA
receptor	NN	O	I-DNA
DNA	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
AhR	NN	O	B-protein
could	NN	O	O
interfere	NN	O	O
with	NN	O	O
BSAP	NN	O	B-protein
-stimulated	NN	O	O
CD19	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
by	NN	O	O
competition	NN	O	O
for	NN	O	O
a	NN	O	O
common	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Inhibitory	NN	O	O
action	NN	O	O
of	NN	O	O
nm23	NN	O	B-protein
proteins	NN	O	I-protein
on	NN	O	O
induction	NN	O	O
of	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
recently	NN	O	O
identified	NN	O	O
a	NN	O	O
differentiation	NN	O	B-protein
inhibitory	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
I-factor	NN	O	B-protein
)	NN	O	O
in	NN	O	O
mouse	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
M1	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
a	NN	O	O
murine	NN	O	B-protein
homolog	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
nm23-H2	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
.	NN	O	O

nm23	NN	O	B-DNA
genes	NN	O	I-DNA
encode	NN	O	O
proteins	NN	O	O
that	NN	O	O
participate	NN	O	O
in	NN	O	O
tumor	NN	O	O
metastasis	NN	O	O
regulation	NN	O	O
and	NN	O	O
in	NN	O	O
various	NN	O	O
fundamental	NN	O	O
cellular	NN	O	O
processes	NN	O	O
,	NN	O	O
although	NN	O	O
their	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
action	NN	O	O
are	NN	O	O
still	NN	O	O
unknown	NN	O	O
.	NN	O	O

Although	NN	O	O
all	NN	O	O
nm23	NN	O	B-protein
proteins	NN	O	I-protein
contain	NN	O	O
nucleoside	NN	O	B-protein
diphosphate	NN	O	I-protein
(	NN	O	I-protein
NDP	NN	O	I-protein
)	NN	O	I-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
established	NN	O	O
that	NN	O	O
the	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
mediated	NN	O	O
the	NN	O	O
various	NN	O	O
functions	NN	O	O
of	NN	O	O
nm23	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
experiment	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
nm23	NN	O	B-protein
proteins	NN	O	I-protein
on	NN	O	O
various	NN	O	O
differentiation	NN	O	O
induction	NN	O	O
systems	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
including	NN	O	O
HL-60	NN	O	B-cell_line
,	NN	O	I-cell_line
U937	NN	O	I-cell_line
,	NN	O	I-cell_line
HEL/S	NN	O	I-cell_line
,	NN	O	I-cell_line
KU812F	NN	O	I-cell_line
,	NN	O	I-cell_line
K562	NN	O	I-cell_line
,	NN	O	I-cell_line
and	NN	O	I-cell_line
HEL	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Native	NN	O	B-protein
human	NN	O	I-protein
erythrocyte	NN	O	I-protein
NDP	NN	O	I-protein
kinase	NN	O	I-protein
protein	NN	O	I-protein
inhibited	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HEL	NN	O	B-cell_line
,	NN	O	I-cell_line
KU812	NN	O	I-cell_line
and	NN	O	I-cell_line
K562	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
monocytic	NN	O	O
or	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
,	NN	O	I-cell_line
U937	NN	O	I-cell_line
and	NN	O	I-cell_line
HEL/S	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HEL	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
recombinant	NN	O	B-protein
human	NN	O	I-protein
nm23-H1	NN	O	I-protein
,	NN	O	I-protein
-H2	NN	O	I-protein
,	NN	O	I-protein
mouse	NN	O	I-protein
nm23-M1	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
-M2	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
both	NN	O	O
the	NN	O	O
mutant	NN	O	B-protein
nm23-H2His	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
truncated	NN	O	B-protein
nm23-H2	NN	O	I-protein
protein	NN	O	I-protein
containing	NN	O	O
N-terminal	NN	O	B-protein
(	NN	O	I-protein
1-60	NN	O	I-protein
)	NN	O	I-protein
peptide	NN	O	I-protein
,	NN	O	O
which	NN	O	O
do	NN	O	O
not	NN	O	O
have	NN	O	O
NDP	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
also	NN	O	O
inhibited	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HEL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
differentiation	NN	O	O
inhibitory	NN	O	O
activity	NN	O	O
of	NN	O	O
I-factor/nm23	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
not	NN	O	O
restricted	NN	O	O
to	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
M1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
activity	NN	O	O
is	NN	O	O
exhibited	NN	O	O
without	NN	O	O
species	NN	O	O
specificity	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
the	NN	O	O
differentiation	NN	O	O
inhibitory	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
nm23/NDP	NN	O	B-protein
kinase	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
independent	NN	O	O
of	NN	O	O
its	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
and	NN	O	O
requires	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
N-terminal	NN	O	O
peptides	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lipopolysaccharide-induced	NN	O	O
E-selectin	NN	O	B-protein
expression	NN	O	O
requires	NN	O	O
continuous	NN	O	O
presence	NN	O	O
of	NN	O	O
LPS	NN	O	O
and	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
bactericidal/permeability-increasing	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
LPS	NN	O	O
express	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
which	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
mediating	NN	O	O
neutrophil	NN	O	B-cell_type
adhesion	NN	O	O
during	NN	O	O
inflammation	NN	O	O
.	NN	O	O

E-selectin	NN	O	B-protein
is	NN	O	O
induced	NN	O	O
within	NN	O	O
1-2	NN	O	O
h	NN	O	O
,	NN	O	O
peaks	NN	O	O
at	NN	O	O
4-6	NN	O	O
h	NN	O	O
,	NN	O	O
and	NN	O	O
gradually	NN	O	O
returns	NN	O	O
to	NN	O	O
basal	NN	O	O
level	NN	O	O
by	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

rBPI21	NN	O	B-protein
,	NN	O	O
a	NN	O	O
recombinant	NN	O	B-protein
N-terminal	NN	O	I-protein
fragment	NN	O	I-protein
of	NN	O	O
human	NN	O	B-protein
bactericidal/permeability-increasing	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
BPI	NN	O	B-protein
)	NN	O	O
,	NN	O	O
inhibited	NN	O	O
LPS-induced	NN	O	O
E-selectin	NN	O	B-protein
expression	NN	O	O
when	NN	O	O
added	NN	O	O
at	NN	O	O
the	NN	O	O
same	NN	O	O
time	NN	O	O
as	NN	O	O
,	NN	O	O
and	NN	O	O
up	NN	O	O
to	NN	O	O
6	NN	O	O
h	NN	O	O
after	NN	O	O
,	NN	O	O
LPS	NN	O	O
.	NN	O	O

Delayed	NN	O	O
administration	NN	O	O
of	NN	O	O
rBPI21	NN	O	B-protein
also	NN	O	O
affected	NN	O	O
LPS-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Two	NN	O	O
to	NN	O	O
4	NN	O	O
h	NN	O	O
following	NN	O	O
LPS	NN	O	O
addition	NN	O	O
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
when	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
already	NN	O	O
activated	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
rBPI21	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
marked	NN	O	O
reduction	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
detectable	NN	O	O
at	NN	O	O
4	NN	O	O
or	NN	O	O
6	NN	O	O
h	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
endothelial	NN	O	O
activation	NN	O	O
requires	NN	O	O
continuous	NN	O	O
presence	NN	O	O
of	NN	O	O
LPS	NN	O	O
,	NN	O	O
and	NN	O	O
rBPI21	NN	O	B-protein
acts	NN	O	O
to	NN	O	O
reverse	NN	O	O
LPS-mediated	NN	O	O
endothelial	NN	O	O
activation	NN	O	O
by	NN	O	O
interrupting	NN	O	O
the	NN	O	O
on-going	NN	O	O
LPS	NN	O	O
signal	NN	O	O
.	NN	O	O

-DOCSTART-	O

GM-CSF	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
share	NN	O	O
common	NN	O	O
control	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Antigen	NN	O	O
complexed	NN	O	O
with	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
class	NN	O	I-protein
I	NN	O	I-protein
or	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
antigen	NN	O	B-cell_type
presenting	NN	O	I-cell_type
cells	NN	O	I-cell_type
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
initiates	NN	O	O
an	NN	O	O
activation	NN	O	O
cascade	NN	O	O
.	NN	O	O

So	NN	O	O
called	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
,	NN	O	O
mediated	NN	O	O
by	NN	O	O
other	NN	O	O
cell	NN	O	O
surface	NN	O	O
interactions	NN	O	O
or	NN	O	O
soluble	NN	O	B-protein
cytokines	NN	O	I-protein
produced	NN	O	O
by	NN	O	O
antigen	NN	O	B-cell_type
presenting	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
are	NN	O	O
also	NN	O	O
required	NN	O	O
for	NN	O	O
complete	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

High	NN	O	O
levels	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
also	NN	O	O
required	NN	O	O
both	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
.	NN	O	O

The	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
requires	NN	O	O
sequences	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
powerful	NN	O	O
enhancer	NN	O	B-DNA
located	NN	O	O
3kb	NN	O	O
upstream	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
-like	NN	O	O
signals	NN	O	O
.	NN	O	O

These	NN	O	O
promoter	NN	O	B-DNA
and	NN	O	I-DNA
enhancer	NN	O	I-DNA
regions	NN	O	I-DNA
are	NN	O	O
mainly	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
by	NN	O	O
TCR	NN	O	B-protein
signals	NN	O	O
has	NN	O	O
been	NN	O	O
well	NN	O	O
described	NN	O	O
for	NN	O	O
interleukin-2	NN	O	O
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IL-4	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Costimulatory	NN	O	O
signals	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
molecule	NN	O	I-protein
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
lead	NN	O	O
to	NN	O	O
activation	NN	O	O
through	NN	O	O
a	NN	O	O
distinct	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
granulocyte-macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
GM-CSF	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
region	NN	O	O
is	NN	O	O
termed	NN	O	O
the	NN	O	O
CK-1	NN	O	B-DNA
or	NN	O	O
CD28RE	NN	O	B-DNA
and	NN	O	O
appears	NN	O	O
to	NN	O	O
bind	NN	O	O
specific	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Human	NN	O	O
T	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
infects	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
can	NN	O	O
lead	NN	O	O
to	NN	O	O
increase	NN	O	O
GM-CSF	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
HTLV-1	NN	O	B-protein
transactivator	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
tax	NN	O	B-protein
,	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
costimulatory	NN	O	O
signal	NN	O	O
for	NN	O	O
GM-CSF	NN	O	O
and	NN	O	O
IL-2	NN	O	O
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
in	NN	O	O
that	NN	O	O
it	NN	O	O
can	NN	O	O
cooperate	NN	O	O
with	NN	O	O
TCR	NN	O	B-protein
signals	NN	O	O
to	NN	O	O
mediate	NN	O	O
high	NN	O	O
level	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Tax	NN	O	O
activates	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
through	NN	O	O
the	NN	O	O
CK-1	NN	O	B-DNA
/CD28RE	NN	O	B-DNA
region	NN	O	O
and	NN	O	O
also	NN	O	O
activates	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
this	NN	O	O
region	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
or	NN	O	O
coactivators	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
tax	NN	O	B-protein
activation	NN	O	O
but	NN	O	O
these	NN	O	O
remain	NN	O	O
to	NN	O	O
be	NN	O	O
identified	NN	O	O
.	NN	O	O

The	NN	O	O
CK-1	NN	O	B-DNA
/CD28RE	NN	O	B-DNA
of	NN	O	O
GM-CSF	NN	O	B-protein
shows	NN	O	O
a	NN	O	O
high	NN	O	O
degree	NN	O	O
of	NN	O	O
similarity	NN	O	O
to	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
CD28RE	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
gene	NN	O	I-DNA
also	NN	O	O
contains	NN	O	O
a	NN	O	O
related	NN	O	O
region	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
both	NN	O	O
GM-CSF	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
respond	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
signals	NN	O	O
via	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
implies	NN	O	O
a	NN	O	O
high	NN	O	O
degree	NN	O	O
of	NN	O	O
conservation	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Danazol	NN	O	O
decreases	NN	O	O
transcription	NN	O	O
of	NN	O	O
estrogen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

1.	NN	O	O
Administration	NN	O	O
of	NN	O	O
danazol	NN	O	O
for	NN	O	O
over	NN	O	O
one	NN	O	O
month	NN	O	O
reduced	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
and	NN	O	O
its	NN	O	O
mRNA	NN	O	B-RNA
to	NN	O	O
approximately	NN	O	O
50	NN	O	O
and	NN	O	O
20	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

2.	NN	O	O
Danazol	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
degradation	NN	O	O
rate	NN	O	O
of	NN	O	O
ER	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

3.	NN	O	O
Danazol	NN	O	O
decreased	NN	O	O
the	NN	O	O
transcription	NN	O	O
rate	NN	O	O
of	NN	O	O
ER	NN	O	B-DNA
gene	NN	O	I-DNA
to	NN	O	O
approximately	NN	O	O
50	NN	O	O
%	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
in	NN	O	O
a	NN	O	O
run-on	NN	O	O
assay	NN	O	O
.	NN	O	O

4.	NN	O	O
Danazol	NN	O	O
may	NN	O	O
release	NN	O	O
estrogen	NN	O	O
predominance	NN	O	O
via	NN	O	O
the	NN	O	O
reduction	NN	O	O
of	NN	O	O
transcription	NN	O	O
for	NN	O	O
ER	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
reduction	NN	O	O
of	NN	O	O
ER	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
ER	NN	O	B-protein
expressions	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
characterization	NN	O	O
of	NN	O	O
novel	NN	O	O
IL-2	NN	O	B-protein
transcriptional	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

IL-2	NN	O	B-protein
-mediated	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
early	NN	O	O
event	NN	O	O
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
process	NN	O	O
.	NN	O	O

Formation	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT-1	NN	O	B-protein
transcriptional	NN	O	I-protein
complex	NN	O	I-protein
on	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
cell	NN	O	O
line	NN	O	O
that	NN	O	O
is	NN	O	O
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
trimer	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT-1	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
a	NN	O	O
lac-Z	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
screened	NN	O	O
for	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
-1-mediated	NN	O	O
beta-galactosidase	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

WIN	NN	O	O
61058	NN	O	O
and	NN	O	O
WIN	NN	O	O
53071	NN	O	O
were	NN	O	O
identified	NN	O	O
as	NN	O	O
microM	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

These	NN	O	O
compounds	NN	O	O
also	NN	O	O
inhibited	NN	O	O
beta-galactosidase	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

Similar	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
-1-mediated	NN	O	O
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
a	NN	O	O
second	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
is	NN	O	O
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
NFAT-1	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
for	NN	O	I-DNA
sCD8	NN	O	I-DNA
.	NN	O	O

At	NN	O	O
10	NN	O	O
microM	NN	O	O
,	NN	O	O
both	NN	O	O
compounds	NN	O	O
inhibited	NN	O	O
IL-2	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
NFAT-1-linked	NN	O	B-cell_line
lac-Z	NN	O	I-cell_line
transfectants	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Both	NN	O	O
compounds	NN	O	O
inhibited	NN	O	O
the	NN	O	O
mixed	NN	O	O
lymphocyte	NN	O	O
reaction	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
inhibition	NN	O	O
was	NN	O	O
reversed	NN	O	O
by	NN	O	O
exogenous	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

WIN	NN	O	O
53071	NN	O	O
inhibited	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
induced	NN	O	O
in	NN	O	O
the	NN	O	O
calcium-dependent	NN	O	O
PMA	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
pathway	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
calcium-independent	NN	O	B-protein
anti-CD28	NN	O	I-protein
Ab	NN	O	I-protein
and	NN	O	O
PMA-induced	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
was	NN	O	O
resistant	NN	O	O
.	NN	O	O

Both	NN	O	O
compounds	NN	O	O
altered	NN	O	O
the	NN	O	O
NFAT-1	NN	O	B-protein
transcriptional	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
causing	NN	O	O
its	NN	O	O
retarded	NN	O	O
mobility	NN	O	O
on	NN	O	O
gels	NN	O	O
.	NN	O	O

By	NN	O	O
these	NN	O	O
functional	NN	O	O
criteria	NN	O	O
,	NN	O	O
we	NN	O	O
believe	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
two	NN	O	O
structurally	NN	O	O
distinct	NN	O	O
,	NN	O	O
novel	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
-1-mediated	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
vacuolar	NN	O	B-DNA
H	NN	O	I-DNA
(	NN	O	I-DNA
+	NN	O	I-DNA
)	NN	O	I-DNA
-ATPase	NN	O	I-DNA
B2	NN	O	I-DNA
subunit	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
differentiating	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Monocyte-macrophage	NN	O	O
differentiation	NN	O	O
was	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
model	NN	O	O
system	NN	O	O
for	NN	O	O
studying	NN	O	O
gene	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
vacuolar	NN	O	I-protein
H	NN	O	I-protein
(	NN	O	I-protein
+	NN	O	I-protein
)	NN	O	I-protein
-ATPase	NN	O	I-protein
(	NN	O	O
V-ATPase	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
mRNA	NN	O	B-RNA
levels	NN	O	O
of	NN	O	O
various	NN	O	O
V-ATPase	NN	O	B-protein
subunits	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
both	NN	O	O
native	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
post-transcriptional	NN	O	O
mechanisms	NN	O	O
could	NN	O	O
account	NN	O	O
for	NN	O	O
increases	NN	O	O
in	NN	O	O
cell	NN	O	O
V-ATPase	NN	O	B-protein
content	NN	O	O
.	NN	O	O

From	NN	O	O
nuclear	NN	O	O
runoff	NN	O	O
experiments	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
one	NN	O	O
subunit	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
the	NN	O	O
B2	NN	O	B-protein
isoform	NN	O	I-protein
(	NN	O	O
Mr	NN	O	O
=	NN	O	O
56	NN	O	O
,	NN	O	O
000	NN	O	O
)	NN	O	O
,	NN	O	O
was	NN	O	O
amplified	NN	O	O
primarily	NN	O	O
by	NN	O	O
transcriptional	NN	O	O
means	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
begun	NN	O	O
to	NN	O	O
examine	NN	O	O
the	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
B2	NN	O	B-DNA
subunit	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Isolation	NN	O	O
and	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	B-DNA
exon	NN	O	I-DNA
and	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
this	NN	O	O
gene	NN	O	O
reveal	NN	O	O
a	NN	O	O
TATA-less	NN	O	B-DNA
promoter	NN	O	I-DNA
with	NN	O	O
a	NN	O	O
high	NN	O	O
G	NN	O	O
+	NN	O	O
C	NN	O	O
content	NN	O	O
.	NN	O	O

Primer	NN	O	O
extension	NN	O	O
and	NN	O	O
ribonuclease	NN	O	B-protein
protection	NN	O	O
analyses	NN	O	O
indicate	NN	O	O
a	NN	O	O
single	NN	O	O
major	NN	O	B-DNA
transcriptional	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
transfected	NN	O	O
promoter-luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
plasmids	NN	O	I-DNA
into	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
define	NN	O	O
sequences	NN	O	O
that	NN	O	O
mediate	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
during	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
sequences	NN	O	O
downstream	NN	O	O
from	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
start	NN	O	O
site	NN	O	O
were	NN	O	O
sufficient	NN	O	O
to	NN	O	O
confer	NN	O	O
increased	NN	O	O
expression	NN	O	O
during	NN	O	O
THP-1	NN	O	B-cell_line
differentiation	NN	O	O
.	NN	O	O

DNase	NN	O	B-protein
I	NN	O	I-protein
footprinting	NN	O	O
and	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
multiple	NN	O	O
AP2	NN	O	B-DNA
and	NN	O	I-DNA
Sp1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
5'-untranslated	NN	O	B-DNA
and	NN	O	I-DNA
proximal	NN	O	I-DNA
coding	NN	O	I-DNA
regions	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Synergy	NN	O	O
between	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
is	NN	O	O
obligatory	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
in	NN	O	O
B	NN	O	B-cell_line
and	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
:	NN	O	O
implication	NN	O	O
for	NN	O	O
c-fos	NN	O	B-DNA
control	NN	O	O
via	NN	O	O
surface	NN	O	B-protein
immunoglobulin	NN	O	I-protein
and	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
protooncogene	NN	O	B-DNA
c-fos	NN	O	I-DNA
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
three	NN	O	O
main	NN	O	O
regulatory	NN	O	O
pathways	NN	O	O
involving	NN	O	O
kinase	NN	O	B-protein
C	NN	O	I-protein
,	NN	O	O
cAMP	NN	O	O
,	NN	O	O
and	NN	O	O
calcium	NN	O	O
.	NN	O	O

Kinase	NN	O	B-protein
C	NN	O	I-protein
mediates	NN	O	O
its	NN	O	O
effects	NN	O	O
via	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
serum	NN	O	B-protein
response	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
SRF	NN	O	B-protein
)	NN	O	O
which	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
serum	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
SRE	NN	O	B-DNA
)	NN	O	O
;	NN	O	O
cAMP	NN	O	O
and	NN	O	O
calcium	NN	O	O
mediate	NN	O	O
their	NN	O	O
effects	NN	O	O
via	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
(	NN	O	O
cAMP	NN	O	B-protein
regulatory	NN	O	I-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
presumably	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
or	NN	O	O
calmodulin-regulated	NN	O	B-protein
kinase	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
these	NN	O	O
elements	NN	O	O
in	NN	O	O
Burkitt	NN	O	B-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
Ramos	NN	O	B-cell_line
and	NN	O	O
Daudi	NN	O	B-cell_line
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
T	NN	O	B-cell_line
lymphocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
Jurkat	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
any	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
pathways	NN	O	O
alone	NN	O	O
has	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
c-fos	NN	O	B-DNA
induction	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
kinase	NN	O	B-protein
C	NN	O	I-protein
activation	NN	O	O
(	NN	O	O
PMA	NN	O	O
stimulation	NN	O	O
)	NN	O	O
combined	NN	O	O
with	NN	O	O
either	NN	O	O
cAMP	NN	O	O
(	NN	O	O
forskolin	NN	O	O
plus	NN	O	O
MIX	NN	O	O
)	NN	O	O
or	NN	O	O
calcium	NN	O	O
stimulation	NN	O	O
(	NN	O	O
ionophore	NN	O	O
)	NN	O	O
leads	NN	O	O
to	NN	O	O
greatly	NN	O	O
enhanced	NN	O	O
c-fos	NN	O	B-DNA
induction	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
cAMP	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
calcium	NN	O	O
shows	NN	O	O
no	NN	O	O
synergy	NN	O	O
in	NN	O	O
c-fos	NN	O	B-DNA
induction	NN	O	O
.	NN	O	O

Okadaic	NN	O	O
acid	NN	O	O
augments	NN	O	O
PMA-	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
calcium-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
has	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
when	NN	O	O
combined	NN	O	O
with	NN	O	O
cAMP	NN	O	O
.	NN	O	O

The	NN	O	O
main	NN	O	O
difference	NN	O	O
between	NN	O	O
Ramos	NN	O	B-cell_line
(	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
and	NN	O	O
Jurkat	NN	O	B-cell_line
(	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
is	NN	O	O
that	NN	O	O
cAMP	NN	O	O
plus	NN	O	O
calcium	NN	O	O
is	NN	O	O
strongly	NN	O	O
synergistic	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
and	NN	O	O
is	NN	O	O
without	NN	O	O
effect	NN	O	O
in	NN	O	O
Ramos	NN	O	B-cell_line
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
using	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
confirms	NN	O	O
that	NN	O	O
the	NN	O	O
requirements	NN	O	O
for	NN	O	O
synergy	NN	O	O
in	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
are	NN	O	O
paralleled	NN	O	O
by	NN	O	O
requirements	NN	O	O
for	NN	O	O
synergy	NN	O	O
in	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Signaling	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
due	NN	O	O
to	NN	O	O
anti-Ig	NN	O	B-protein
stimulation	NN	O	O
involves	NN	O	O
both	NN	O	O
kinase	NN	O	B-protein
C	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
release	NN	O	O
of	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
,	NN	O	O
and	NN	O	O
results	NN	O	O
in	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
synergy	NN	O	O
between	NN	O	O
the	NN	O	O
kinase	NN	O	B-protein
C	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
calcium	NN	O	O
is	NN	O	O
needed	NN	O	O
for	NN	O	O
efficient	NN	O	O
c-fos	NN	O	B-DNA
induction	NN	O	O
since	NN	O	O
neither	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
alone	NN	O	O
induces	NN	O	O
c-fos	NN	O	B-DNA
well	NN	O	O
.	NN	O	O

That	NN	O	O
synergy	NN	O	O
of	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
is	NN	O	O
relevant	NN	O	O
for	NN	O	O
the	NN	O	O
anti-Ig	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
is	NN	O	O
supported	NN	O	O
by	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
cAMP-inducing	NN	O	O
agents	NN	O	O
and	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
further	NN	O	O
enhance	NN	O	O
anti-Ig	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
cell-specific	NN	O	O
patterns	NN	O	O
of	NN	O	O
synergy	NN	O	O
are	NN	O	O
an	NN	O	O
essential	NN	O	O
feature	NN	O	O
for	NN	O	O
c-fos	NN	O	B-DNA
induction	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
relevant	NN	O	O
for	NN	O	O
c-fos	NN	O	B-DNA
control	NN	O	O
through	NN	O	O
B	NN	O	B-protein
and	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Multiple	NN	O	O
signals	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
granulocyte-macrophage	NN	O	I-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
granulocyte-macrophage	NN	O	I-DNA
CSF	NN	O	I-DNA
(	NN	O	I-DNA
GM-CSF	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
activation	NN	O	O
that	NN	O	O
can	NN	O	O
be	NN	O	O
mimicked	NN	O	O
by	NN	O	O
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
PMA	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
contains	NN	O	O
a	NN	O	O
proximal	NN	O	B-DNA
functional	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
-620	NN	O	B-DNA
to	NN	O	I-DNA
+34	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
powerful	NN	O	O
enhancer	NN	O	B-DNA
located	NN	O	O
3	NN	O	O
kb	NN	O	O
upstream	NN	O	O
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

The	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
a	NN	O	O
region	NN	O	O
termed	NN	O	O
CLEO	NN	O	B-DNA
(	NN	O	O
-54	NN	O	B-DNA
to	NN	O	I-DNA
-31	NN	O	I-DNA
)	NN	O	O
that	NN	O	O
consists	NN	O	O
of	NN	O	O
a	NN	O	O
purine-rich	NN	O	B-DNA
element	NN	O	I-DNA
abutting	NN	O	O
an	NN	O	O
activator	NN	O	B-DNA
protein-1	NN	O	I-DNA
(	NN	O	I-DNA
AP-1	NN	O	I-DNA
)	NN	O	I-DNA
-like	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
an	NN	O	O
upstream	NN	O	B-DNA
nuclear	NN	O	I-DNA
factor-kappa	NN	O	I-DNA
B	NN	O	I-DNA
(	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
-85	NN	O	B-DNA
to	NN	O	I-DNA
-76	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
a	NN	O	O
CK-1	NN	O	B-DNA
element	NN	O	I-DNA
(	NN	O	O
-101	NN	O	B-DNA
to	NN	O	I-DNA
-92	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
in	NN	O	O
this	NN	O	O
work	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
either	NN	O	O
the	NN	O	O
purine-rich	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CLEO	NN	O	B-DNA
element	NN	O	I-DNA
or	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
result	NN	O	O
in	NN	O	O
reduced	NN	O	O
PMA/Ca2+	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
620-bp	NN	O	B-DNA
human	NN	O	I-DNA
GM-CSF	NN	O	I-DNA
promoter-luciferase	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
65	NN	O	O
%	NN	O	O
and	NN	O	O
50	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
major	NN	O	O
inducible	NN	O	B-protein
protein	NN	O	I-protein
complex	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CLEO	NN	O	I-DNA
(	NN	O	I-DNA
hCLEO	NN	O	I-DNA
)	NN	O	I-DNA
element	NN	O	I-DNA
is	NN	O	O
an	NN	O	O
AP-1-like	NN	O	B-protein
complex	NN	O	I-protein
that	NN	O	O
is	NN	O	O
inducible	NN	O	O
by	NN	O	O
PMA	NN	O	O
alone	NN	O	O
,	NN	O	O
but	NN	O	O
shows	NN	O	O
increased	NN	O	O
binding	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
PMA	NN	O	O
together	NN	O	O
with	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
is	NN	O	O
not	NN	O	O
cyclosporin-sensitive	NN	O	O
,	NN	O	O
promoter	NN	O	O
induction	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
treatment	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
weak	NN	O	O
inducible	NN	O	O
complex	NN	O	O
resembling	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
hCLEO	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

By	NN	O	O
using	NN	O	O
recombinant	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
we	NN	O	O
confirmed	NN	O	O
that	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NF-ATp	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
higher	NN	O	O
order	NN	O	O
NF-ATp/AP-1	NN	O	B-protein
complex	NN	O	I-protein
could	NN	O	O
all	NN	O	O
form	NN	O	O
with	NN	O	O
the	NN	O	O
hCLEO	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
we	NN	O	O
have	NN	O	O
also	NN	O	O
defined	NN	O	O
the	NN	O	O
sequence	NN	O	O
requirements	NN	O	O
for	NN	O	O
binding	NN	O	O
of	NN	O	O
each	NN	O	O
of	NN	O	O
these	NN	O	O
complexes	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
constitutively	NN	O	O
active	NN	O	O
form	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
could	NN	O	O
substitute	NN	O	O
for	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
and	NN	O	O
synergize	NN	O	O
with	NN	O	O
PMA	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
conversely	NN	O	O
that	NN	O	O
mutant-activated	NN	O	B-protein
Ras	NN	O	I-protein
could	NN	O	O
substitute	NN	O	O
for	NN	O	O
PMA	NN	O	O
and	NN	O	O
cooperate	NN	O	O
with	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
.	NN	O	O

Co-expression	NN	O	O
of	NN	O	O
Ras	NN	O	B-protein
and	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
however	NN	O	O
,	NN	O	O
did	NN	O	O
not	NN	O	O
activate	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
required	NN	O	O
the	NN	O	O
additional	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
imply	NN	O	O
that	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
signals	NN	O	O
are	NN	O	O
required	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
proximal	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
signals	NN	O	O
impinge	NN	O	O
on	NN	O	O
distinct	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
hCLEO	NN	O	B-DNA
and	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

IL-10	NN	O	O
induces	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
tyk2	NN	O	B-protein
and	NN	O	O
Jak1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
differential	NN	O	O
assembly	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
STAT3	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

IL-10	NN	O	B-protein
affects	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
driving	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
immune	NN	O	O
responsiveness	NN	O	O
such	NN	O	O
that	NN	O	O
Th2	NN	O	O
lymphocyte-mediated	NN	O	O
effects	NN	O	O
predominate	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
IL-10	NN	O	B-protein
stimulates	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	O
activators	NN	O	B-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
,	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
in	NN	O	O
a	NN	O	O
differential	NN	O	O
manner	NN	O	O
such	NN	O	O
that	NN	O	O
the	NN	O	O
relative	NN	O	O
formation	NN	O	O
of	NN	O	O
homo-	NN	O	B-protein
and	NN	O	I-protein
heterodimers	NN	O	I-protein
varies	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
monocytes	NN	O	B-cell_type
express	NN	O	O
a	NN	O	O
novel	NN	O	O
IL-10-stimulated	NN	O	B-protein
STAT	NN	O	I-protein
protein	NN	O	I-protein
with	NN	O	O
an	NN	O	O
M	NN	O	O
(	NN	O	O
r	NN	O	O
)	NN	O	O
of	NN	O	O
70	NN	O	B-protein
kDa	NN	O	I-protein
that	NN	O	O
is	NN	O	O
recognized	NN	O	O
by	NN	O	O
the	NN	O	O
anti-STAT3	NN	O	B-protein
Ab	NN	O	I-protein
but	NN	O	O
is	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

IL-10	NN	O	B-protein
treatment	NN	O	O
of	NN	O	O
both	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
results	NN	O	O
in	NN	O	O
the	NN	O	O
ligand-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
tyk2	NN	O	B-protein
and	NN	O	O
Jak1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Jak2	NN	O	B-protein
or	NN	O	O
Jak3	NN	O	B-protein
.	NN	O	O

Selective	NN	O	O
modulation	NN	O	O
of	NN	O	O
immune	NN	O	O
responsiveness	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
in	NN	O	O
cells	NN	O	O
such	NN	O	O
as	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
result	NN	O	O
in	NN	O	O
part	NN	O	O
from	NN	O	O
the	NN	O	O
differential	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
protein	NN	O	I-protein
pairs	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Use	NN	O	O
of	NN	O	O
new	NN	O	O
biologic	NN	O	O
markers	NN	O	O
in	NN	O	O
the	NN	O	O
ovulation	NN	O	O
induction	NN	O	O
.	NN	O	O

Biological	NN	O	O
markers	NN	O	O
of	NN	O	O
ovulation	NN	O	O
,	NN	O	O
after	NN	O	O
a	NN	O	O
great	NN	O	O
in	NN	O	O
the	NN	O	O
past	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
fallen	NN	O	O
into	NN	O	O
disuse	NN	O	O
for	NN	O	O
the	NN	O	O
large	NN	O	O
diffusion	NN	O	O
of	NN	O	O
biochemical	NN	O	O
and	NN	O	O
biophysical	NN	O	O
ones	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
real	NN	O	O
effect	NN	O	O
of	NN	O	O
hormones	NN	O	O
involved	NN	O	O
in	NN	O	O
ovulation	NN	O	O
is	NN	O	O
expressed	NN	O	O
by	NN	O	O
biological	NN	O	O
modifications	NN	O	O
on	NN	O	O
target	NN	O	B-cell_type
tissues	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
explore	NN	O	O
the	NN	O	O
modifications	NN	O	O
of	NN	O	O
not	NN	O	O
reproductive	NN	O	O
target	NN	O	O
tissues	NN	O	O
as	NN	O	O
ovulation	NN	O	O
markers	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
behaviour	NN	O	O
of	NN	O	O
Albuminemia	NN	O	O
,	NN	O	O
Platelet	NN	O	B-protein
Factor	NN	O	I-protein
IV	NN	O	I-protein
(	NN	O	O
as	NN	O	O
indicator	NN	O	O
of	NN	O	O
Platelet	NN	O	O
Aggregation	NN	O	O
)	NN	O	O
,	NN	O	O
Type	NN	O	B-protein
II	NN	O	I-protein
estrogenic	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
42	NN	O	O
ovulation	NN	O	O
induced	NN	O	O
women	NN	O	O
,	NN	O	O
undergoing	NN	O	O
our	NN	O	O
observation	NN	O	O
.	NN	O	O

33	NN	O	O
of	NN	O	O
them	NN	O	O
had	NN	O	O
ovulation	NN	O	O
and	NN	O	O
9	NN	O	O
developed	NN	O	O
a	NN	O	O
LUF	NN	O	O
syndrome	NN	O	O
,	NN	O	O
constituting	NN	O	O
two	NN	O	O
biological	NN	O	O
models	NN	O	O
of	NN	O	O
an	NN	O	O
opposite	NN	O	O
situation	NN	O	O
for	NN	O	O
the	NN	O	O
three	NN	O	O
markers	NN	O	O
observed	NN	O	O
.	NN	O	O

All	NN	O	O
the	NN	O	O
markers	NN	O	O
considered	NN	O	O
were	NN	O	O
sufficiently	NN	O	O
sensitive	NN	O	O
,	NN	O	O
but	NN	O	O
among	NN	O	O
them	NN	O	O
,	NN	O	O
Platelet	NN	O	B-protein
Factor	NN	O	I-protein
IV	NN	O	I-protein
was	NN	O	O
the	NN	O	O
most	NN	O	O
reliable	NN	O	O
to	NN	O	O
the	NN	O	O
hormonal	NN	O	O
ovulatory	NN	O	O
situation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Abnormal	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
lpr	NN	O	O
CD4-CD8-	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
results	NN	O	O
in	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells-binding	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
inert	NN	O	O
quality	NN	O	O
of	NN	O	O
MRL-Ipr/Ipr	NN	O	B-cell_line
(	NN	O	I-cell_line
Ipr	NN	O	I-cell_line
)	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
CD4-CD8-	NN	O	I-cell_line
(	NN	O	I-cell_line
CD4-8-	NN	O	I-cell_line
)	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
manifests	NN	O	O
primarily	NN	O	O
as	NN	O	O
an	NN	O	O
inability	NN	O	O
to	NN	O	O
proliferate	NN	O	O
or	NN	O	O
produce	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
or	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Yet	NN	O	O
these	NN	O	O
same	NN	O	O
cells	NN	O	O
do	NN	O	O
initiate	NN	O	O
early	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
events	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
generation	NN	O	O
of	NN	O	O
inositol	NN	O	O
phosphates	NN	O	O
and	NN	O	O
increased	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
.	NN	O	O

They	NN	O	O
also	NN	O	O
display	NN	O	O
constitutively	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
p59fyn	NN	O	B-protein
and	NN	O	O
CD3	NN	O	B-protein
zeta	NN	O	I-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

The	NN	O	O
generation	NN	O	O
of	NN	O	O
second	NN	O	O
messengers	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
normally	NN	O	O
leads	NN	O	O
to	NN	O	O
downstream	NN	O	O
signaling	NN	O	O
that	NN	O	O
results	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
compared	NN	O	O
the	NN	O	O
activation	NN	O	O
state	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
in	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
and	NN	O	O
stimulated	NN	O	O
Ipr	NN	O	B-cell_line
CD4-8-	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
that	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Levels	NN	O	O
of	NN	O	O
the	NN	O	O
octamer	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
p50-p65	NN	O	B-protein
heterodimer	NN	O	I-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
transcriptional	NN	O	I-protein
factors	NN	O	I-protein
are	NN	O	O
constitutively	NN	O	O
elevated	NN	O	O
in	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
Ipr	NN	O	B-cell_line
CD4-8-	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
activated	NN	O	O
phenotype	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
mitogens	NN	O	O
,	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
transactivating	NN	O	O
complex	NN	O	O
,	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
,	NN	O	O
occurs	NN	O	O
with	NN	O	O
normal	NN	O	O
kinetics	NN	O	O
in	NN	O	O
Ipr	NN	O	B-cell_line
CD4-8-	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Yet	NN	O	O
,	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
activating	NN	O	O
NF-AT	NN	O	B-protein
complex	NN	O	I-protein
never	NN	O	O
reach	NN	O	O
those	NN	O	O
observed	NN	O	O
in	NN	O	O
similarly	NN	O	O
stimulated	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
Ipr	NN	O	B-cell_line
CD4-8-	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
display	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
specific	NN	O	O
binding	NN	O	O
activity	NN	O	O
at	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
present	NN	O	O
at	NN	O	O
much	NN	O	O
lower	NN	O	O
levels	NN	O	O
in	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Upon	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
novel	NN	O	O
NF-AT-binding	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
rapidly	NN	O	O
down-regulated	NN	O	O
in	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
persists	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
in	NN	O	O
Ipr	NN	O	B-cell_line
CD4-8-	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
two	NN	O	O
abnormalities	NN	O	O
at	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-DNA
site	NN	O	I-DNA
provide	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
to	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
defect	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
from	NN	O	O
Ipr	NN	O	B-cell_line
CD4-8-	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Cross-linking	NN	O	O
of	NN	O	O
CD30	NN	O	B-protein
induces	NN	O	O
HIV	NN	O	O
expression	NN	O	O
in	NN	O	O
chronically	NN	O	O
infected	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CD30	NN	O	B-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor	NN	O	O
(	NN	O	O
TNF	NN	O	O
)	NN	O	O
receptor	NN	O	O
family	NN	O	O
,	NN	O	O
is	NN	O	O
expressed	NN	O	O
constitutively	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
line	NN	O	O
ACH-2	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
chronically	NN	O	O
infected	NN	O	O
with	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type-1	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
-1	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
cross-linking	NN	O	O
CD30	NN	O	B-protein
with	NN	O	O
an	NN	O	O
anti-	NN	O	O
CD30	NN	O	B-protein
-specific	NN	O	O
monoclonal	NN	O	O
antibody	NN	O	O
,	NN	O	O
which	NN	O	O
mimics	NN	O	O
the	NN	O	O
described	NN	O	O
biological	NN	O	O
activities	NN	O	O
of	NN	O	O
the	NN	O	O
CD30	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
CD30L	NN	O	B-protein
)	NN	O	O
,	NN	O	O
results	NN	O	O
in	NN	O	O
HIV	NN	O	O
expression	NN	O	O
.	NN	O	O

CD30	NN	O	B-protein
cross-linking	NN	O	O
does	NN	O	O
not	NN	O	O
alter	NN	O	O
proliferation	NN	O	O
of	NN	O	O
ACH-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
is	NN	O	O
not	NN	O	O
mediated	NN	O	O
by	NN	O	O
endogenous	NN	O	O
TNF	NN	O	B-protein
alpha/beta	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
CD30	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
enhanced	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
CD30	NN	O	B-protein
-CD30L	NN	O	B-protein
interactions	NN	O	O
mediate	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
by	NN	O	O
a	NN	O	O
kappa	NN	O	O
B-dependent	NN	O	O
pathway	NN	O	O
that	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
.	NN	O	O

This	NN	O	O
mechanism	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
from	NN	O	O
latently	NN	O	O
infected	NN	O	O
CD4+	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
especially	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
where	NN	O	O
cell	NN	O	O
to	NN	O	O
cell	NN	O	O
contact	NN	O	O
is	NN	O	O
conducive	NN	O	O
to	NN	O	O
receptor-ligand	NN	O	O
interactions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
directed	NN	O	O
by	NN	O	O
the	NN	O	O
carboxyl	NN	O	B-protein
terminus	NN	O	I-protein
of	NN	O	O
CIITA	NN	O	B-DNA
,	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
defective	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
type	NN	O	O
II	NN	O	O
MHC	NN	O	B-protein
combined	NN	O	O
immune	NN	O	O
deficiency	NN	O	O
.	NN	O	O

Type	NN	O	B-protein
II	NN	O	I-protein
major	NN	O	I-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
combined	NN	O	O
immune	NN	O	O
deficiency	NN	O	O
(	NN	O	O
type	NN	O	O
II	NN	O	O
MHC	NN	O	B-protein
CID	NN	O	O
or	NN	O	O
bare	NN	O	O
lymphocyte	NN	O	O
syndrome	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
congenital	NN	O	O
immunodeficiency	NN	O	O
disease	NN	O	O
characterized	NN	O	O
by	NN	O	O
absent	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Four	NN	O	O
distinct	NN	O	O
complementation	NN	O	O
groups	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
the	NN	O	O
defective	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
group	NN	O	O
II	NN	O	O
type	NN	O	O
II	NN	O	O
MHC	NN	O	B-protein
CID	NN	O	O
has	NN	O	O
been	NN	O	O
isolated	NN	O	O
and	NN	O	O
termed	NN	O	O
CIITA	NN	O	B-DNA
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
CIITA	NN	O	B-DNA
is	NN	O	O
an	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene-specific	NN	O	I-DNA
transcription	NN	O	I-DNA
activator	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
transcription	NN	O	O
activation	NN	O	O
function	NN	O	O
is	NN	O	O
provided	NN	O	O
by	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
acidic	NN	O	I-protein
domain	NN	O	I-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
26-137	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
experimentally	NN	O	O
exchangeable	NN	O	O
with	NN	O	O
a	NN	O	O
heterologous	NN	O	B-protein
viral	NN	O	I-protein
transcription-activating	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

The	NN	O	O
specificity	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
for	NN	O	O
three	NN	O	O
major	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
DR	NN	O	B-DNA
,	NN	O	O
DQ	NN	O	B-DNA
and	NN	O	O
DP	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
its	NN	O	O
remaining	NN	O	O
C-terminal	NN	O	B-protein
residues	NN	O	I-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
317-1130	NN	O	I-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
transactivation	NN	O	O
of	NN	O	O
multiple	NN	O	B-DNA
cis	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
especially	NN	O	O
S	NN	O	B-DNA
and	NN	O	O
X2	NN	O	B-DNA
,	NN	O	O
of	NN	O	O
the	NN	O	O
DR	NN	O	B-DNA
alpha	NN	O	I-DNA
proximal	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
group	NN	O	B-cell_line
II	NN	O	I-cell_line
CID	NN	O	I-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
CIITA	NN	O	B-protein
dependent	NN	O	O
.	NN	O	O

Since	NN	O	O
CIITA	NN	O	B-protein
overexpression	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
did	NN	O	O
not	NN	O	O
increase	NN	O	O
class	NN	O	O
II	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
initiation	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
expression	NN	O	O
serves	NN	O	O
as	NN	O	O
the	NN	O	O
on-off	NN	O	O
switch	NN	O	O
,	NN	O	O
while	NN	O	O
availability	NN	O	O
of	NN	O	O
downstream	NN	O	O
interactor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
limits	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Monocyte	NN	O	O
tethering	NN	O	O
by	NN	O	O
P-selectin	NN	O	B-protein
regulates	NN	O	O
monocyte	NN	O	B-protein
chemotactic	NN	O	I-protein
protein-1	NN	O	I-protein
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
secretion	NN	O	O
.	NN	O	O

Signal	NN	O	O
integration	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Adhesion	NN	O	B-protein
molecules	NN	O	I-protein
that	NN	O	O
tether	NN	O	O
circulating	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
also	NN	O	O
transduce	NN	O	O
or	NN	O	O
modulate	NN	O	O
outside-in	NN	O	O
signals	NN	O	O
for	NN	O	O
cellular	NN	O	O
activation	NN	O	O
,	NN	O	O
providing	NN	O	O
an	NN	O	O
initial	NN	O	O
regulatory	NN	O	O
point	NN	O	O
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
.	NN	O	O

Adhesion	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
to	NN	O	O
P-selectin	NN	O	B-protein
,	NN	O	O
the	NN	O	O
most	NN	O	O
rapidly	NN	O	O
expressed	NN	O	O
endothelial	NN	O	B-protein
tethering	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
increased	NN	O	O
the	NN	O	O
secretion	NN	O	O
of	NN	O	O
monocyte	NN	O	B-protein
chemotactic	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
MCP-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
by	NN	O	O
the	NN	O	O
leukocytes	NN	O	B-cell_type
when	NN	O	O
they	NN	O	O
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

Increased	NN	O	O
cytokine	NN	O	O
secretion	NN	O	O
was	NN	O	O
specifically	NN	O	O
inhibited	NN	O	O
by	NN	O	O
G1	NN	O	B-protein
,	NN	O	O
an	NN	O	O
anti-P-selectin	NN	O	B-protein
mAb	NN	O	I-protein
that	NN	O	O
prevents	NN	O	O
P-selectin	NN	O	B-protein
from	NN	O	O
binding	NN	O	O
to	NN	O	O
its	NN	O	O
ligand	NN	O	O
(	NN	O	O
P-selectin	NN	O	B-protein
glycoprotein	NN	O	I-protein
ligand-1	NN	O	I-protein
)	NN	O	O
on	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
tethering	NN	O	O
by	NN	O	O
P-selectin	NN	O	B-protein
specifically	NN	O	O
enhanced	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
required	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
,	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
other	NN	O	O
immediate-early	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
P-selectin	NN	O	B-protein
,	NN	O	O
through	NN	O	O
its	NN	O	O
ligands	NN	O	O
on	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
may	NN	O	O
locally	NN	O	O
regulate	NN	O	O
cytokine	NN	O	B-protein
secretion	NN	O	O
in	NN	O	O
inflamed	NN	O	O
tissues	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
roles	NN	O	O
of	NN	O	O
in	NN	O	B-DNA
vivo	NN	O	I-DNA
footprinted	NN	O	I-DNA
DNA	NN	O	I-DNA
motifs	NN	O	I-DNA
within	NN	O	O
an	NN	O	O
alpha-globin	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Erythroid	NN	O	O
lineage	NN	O	O
and	NN	O	O
developmental	NN	O	O
stage	NN	O	O
specificities	NN	O	O
.	NN	O	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
alpha-like	NN	O	I-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
embryonic	NN	O	B-DNA
zeta	NN	O	I-DNA
2	NN	O	I-DNA
and	NN	O	O
adult	NN	O	B-DNA
alpha	NN	O	I-DNA
,	NN	O	O
during	NN	O	O
erythroid	NN	O	O
development	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
distal	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
HS-40	NN	O	B-DNA
.	NN	O	O

Previous	NN	O	O
protein-DNA	NN	O	O
binding	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
HS-40	NN	O	B-DNA
consists	NN	O	O
of	NN	O	O
multiple	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
occupied	NN	O	O
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
an	NN	O	O
erythroid	NN	O	O
lineage-	NN	O	O
and	NN	O	O
developmental	NN	O	O
stage-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
systematically	NN	O	O
analyzed	NN	O	O
the	NN	O	O
functional	NN	O	O
roles	NN	O	O
of	NN	O	O
these	NN	O	O
factor	NN	O	B-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
of	NN	O	O
HS-40	NN	O	B-DNA
by	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
and	NN	O	O
transient	NN	O	O
expression	NN	O	O
assay	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

Three	NN	O	O
of	NN	O	O
these	NN	O	O
HS-40	NN	O	B-DNA
enhancer	NN	O	I-DNA
motifs	NN	O	I-DNA
,	NN	O	O
5'NF-E2/AP1	NN	O	B-DNA
,	NN	O	O
GT	NN	O	B-DNA
II	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
GATA-1	NN	O	B-DNA
(	NN	O	I-DNA
c	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
positively	NN	O	O
regulate	NN	O	O
the	NN	O	O
zeta	NN	O	B-DNA
2-globin	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
embryonic/fetal	NN	O	B-cell_line
erythroid	NN	O	I-cell_line
K562	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
adult	NN	O	B-DNA
alpha-globin	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
adult	NN	O	B-cell_line
erythroid	NN	O	I-cell_line
MEL	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
the	NN	O	O
3'NF-E2/AP1	NN	O	B-DNA
motif	NN	O	I-DNA
is	NN	O	O
able	NN	O	O
to	NN	O	O
exert	NN	O	O
both	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
zeta	NN	O	B-DNA
2-globin	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
this	NN	O	O
dual	NN	O	O
function	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
modulated	NN	O	O
through	NN	O	O
differential	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
ubiquitous	NN	O	B-protein
AP1	NN	O	I-protein
factors	NN	O	I-protein
and	NN	O	O
the	NN	O	O
erythroid-enriched	NN	O	B-protein
NF-E2	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Mutation	NN	O	O
in	NN	O	O
the	NN	O	O
GATA-1	NN	O	B-DNA
(	NN	O	I-DNA
d	NN	O	I-DNA
)	NN	O	I-DNA
motif	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
exhibits	NN	O	O
an	NN	O	O
adult	NN	O	B-DNA
erythroid-specific	NN	O	I-DNA
genomic	NN	O	I-DNA
footprint	NN	O	I-DNA
,	NN	O	O
decreases	NN	O	O
the	NN	O	O
HS-40	NN	O	B-DNA
enhancer	NN	O	I-DNA
function	NN	O	O
in	NN	O	O
dimethyl	NN	O	B-cell_line
sulfoxide-induced	NN	O	I-cell_line
MEL	NN	O	I-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
studies	NN	O	O
have	NN	O	O
defined	NN	O	O
the	NN	O	O
regulatory	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
different	NN	O	O
HS-40	NN	O	B-DNA
motifs	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
remarkable	NN	O	O
correlation	NN	O	O
between	NN	O	O
genomic	NN	O	O
footprinting	NN	O	O
data	NN	O	O
and	NN	O	O
the	NN	O	O
mutagenesis	NN	O	O
results	NN	O	O
also	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
erythroid	NN	O	O
lineage-	NN	O	O
and	NN	O	O
developmental	NN	O	O
stage-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
alpha-like	NN	O	I-DNA
globin	NN	O	I-DNA
promoters	NN	O	I-DNA
is	NN	O	O
indeed	NN	O	O
modulated	NN	O	O
by	NN	O	O
stable	NN	O	O
binding	NN	O	O
of	NN	O	O
specific	NN	O	B-protein
nuclear	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nitric	NN	O	O
oxide-stimulated	NN	O	O
guanine	NN	O	O
nucleotide	NN	O	O
exchange	NN	O	O
on	NN	O	O
p21ras	NN	O	B-protein
.	NN	O	O

The	NN	O	O
protooncogene	NN	O	B-DNA
p21ras	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
monomeric	NN	O	B-protein
G	NN	O	I-protein
protein	NN	O	I-protein
family	NN	O	I-protein
member	NN	O	I-protein
,	NN	O	O
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
converting	NN	O	O
extracellular	NN	O	O
signals	NN	O	O
into	NN	O	O
intracellular	NN	O	O
biochemical	NN	O	O
events	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
(	NN	O	O
NO	NN	O	O
)	NN	O	O
activates	NN	O	O
p21ras	NN	O	B-DNA
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
GTP-bound	NN	O	B-protein
p21ras	NN	O	I-protein
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
studies	NN	O	O
using	NN	O	O
pure	NN	O	B-protein
recombinant	NN	O	I-protein
p21ras	NN	O	I-protein
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
is	NN	O	O
direct	NN	O	O
and	NN	O	O
reversible	NN	O	O
.	NN	O	O

Circular	NN	O	O
dichroism	NN	O	O
analysis	NN	O	O
reveals	NN	O	O
that	NN	O	O
NO	NN	O	O
induces	NN	O	O
a	NN	O	O
profound	NN	O	O
conformational	NN	O	O
change	NN	O	O
in	NN	O	O
p21ras	NN	O	B-DNA
in	NN	O	O
association	NN	O	O
with	NN	O	O
GDP/GTP	NN	O	O
exchange	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
activation	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
S-nitrosylation	NN	O	O
of	NN	O	O
a	NN	O	O
critical	NN	O	O
cysteine	NN	O	O
residue	NN	O	O
which	NN	O	O
stimulates	NN	O	O
guanine	NN	O	O
nucleotide	NN	O	O
exchange	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
p21ras	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
NO-induced	NN	O	O
downstream	NN	O	O
signaling	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
endogenous	NN	O	O
NO	NN	O	O
can	NN	O	O
activate	NN	O	O
p21ras	NN	O	B-protein
in	NN	O	O
the	NN	O	O
same	NN	O	O
cell	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
identify	NN	O	O
p21ras	NN	O	B-DNA
as	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
cell	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
identify	NN	O	O
p21ras	NN	O	B-DNA
as	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
NO	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
NO	NN	O	O
activates	NN	O	O
p21ras	NN	O	B-DNA
by	NN	O	O
an	NN	O	O
action	NN	O	O
which	NN	O	O
mimics	NN	O	O
that	NN	O	O
of	NN	O	O
guanine	NN	O	B-protein
nucleotide	NN	O	I-protein
exchange	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Interleukin-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus-transformed	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
nuclear	NN	O	B-protein
factor-chi	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
interleukin	NN	O	B-DNA
(	NN	O	I-DNA
IL	NN	O	I-DNA
)	NN	O	I-DNA
-2	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
has	NN	O	O
been	NN	O	O
investigated	NN	O	O
mainly	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
predominant	NN	O	O
producers	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
also	NN	O	O
synthesize	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus	NN	O	I-cell_line
(	NN	O	I-cell_line
EBV	NN	O	I-cell_line
)	NN	O	I-cell_line
-transformed	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
which	NN	O	O
are	NN	O	O
capable	NN	O	O
of	NN	O	O
secreting	NN	O	O
IL-2	NN	O	B-protein
at	NN	O	O
a	NN	O	O
low	NN	O	O
level	NN	O	O
after	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
and	NN	O	O
the	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
ionomycin	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfections	NN	O	O
using	NN	O	O
reporter	NN	O	B-DNA
constructs	NN	O	I-DNA
with	NN	O	O
multiples	NN	O	O
of	NN	O	O
transcription	NN	O	B-DNA
factor	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
[	NN	O	O
distal	NN	O	B-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-AT	NN	O	I-protein
,	NN	O	O
proximal	NN	O	B-protein
NF-AT	NN	O	I-protein
,	NN	O	O
AP-1/Octamer	NN	O	B-protein
(	NN	O	O
UPS	NN	O	B-protein
)	NN	O	O
or	NN	O	O
NF-chi	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
TCEd	NN	O	B-protein
)	NN	O	O
sites	NN	O	O
]	NN	O	O
were	NN	O	O
performed	NN	O	O
.	NN	O	O

In	NN	O	O
EBV-transformed	NN	O	B-cell_line
B	NN	O	I-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
chi	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
exerted	NN	O	O
the	NN	O	O
strongest	NN	O	O
inducible	NN	O	O
activity	NN	O	O
;	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
showed	NN	O	O
either	NN	O	O
no	NN	O	O
or	NN	O	O
only	NN	O	O
weak	NN	O	O
activity	NN	O	O
compared	NN	O	O
to	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

An	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
bearing	NN	O	O
a	NN	O	O
defective	NN	O	O
NF-chi	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
completely	NN	O	O
inactive	NN	O	O
in	NN	O	O
EBV-transformed	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
it	NN	O	O
still	NN	O	O
had	NN	O	O
activity	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
seven	NN	O	O
EBV-B	NN	O	B-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
or	NN	O	O
lines	NN	O	O
differing	NN	O	O
in	NN	O	O
their	NN	O	O
capacity	NN	O	O
to	NN	O	O
secrete	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
correlated	NN	O	O
well	NN	O	O
with	NN	O	O
the	NN	O	O
status	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
whose	NN	O	O
activity	NN	O	O
is	NN	O	O
controlled	NN	O	O
through	NN	O	O
chi	NN	O	B-protein
B	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
active	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-cell_line
producing	NN	O	I-cell_line
EBV-B	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
inactive	NN	O	O
in	NN	O	O
the	NN	O	O
non-IL-2-producing	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
using	NN	O	O
protein	NN	O	O
extracts	NN	O	O
from	NN	O	O
EBV-B	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
NF-chi	NN	O	B-protein
B	NN	O	I-protein
probe	NN	O	O
revealed	NN	O	O
the	NN	O	O
constitutive	NN	O	O
generation	NN	O	O
of	NN	O	O
chi	NN	O	B-protein
B	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
IL-2-secreting	NN	O	B-cell_line
cells	NN	O	I-cell_line
consisting	NN	O	O
mainly	NN	O	O
of	NN	O	O
heterodimeric	NN	O	B-protein
p50/p65	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

A	NN	O	O
weaker	NN	O	O
chi	NN	O	B-DNA
B	NN	O	I-DNA
complex	NN	O	O
formation	NN	O	O
and	NN	O	O
faster-migrating	NN	O	O
complexes	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
non-IL-2-secreting	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
NF-chi	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
is	NN	O	O
indispensable	NN	O	O
for	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
EBV-transformed	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
appear	NN	O	O
to	NN	O	O
be	NN	O	O
less	NN	O	O
important	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Latent	NN	O	B-protein
membrane	NN	O	I-protein
protein-1	NN	O	I-protein
induces	NN	O	O
cyclin	NN	O	B-protein
D2	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
pRb	NN	O	B-protein
hyperphosphorylation	NN	O	O
,	NN	O	O
and	NN	O	O
loss	NN	O	O
of	NN	O	O
TGF-beta	NN	O	B-protein
1	NN	O	I-protein
-mediated	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
in	NN	O	O
EBV-positive	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
normal	NN	O	O
cell	NN	O	O
cycle	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
several	NN	O	O
molecules	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
tumor-suppressor	NN	O	B-protein
protein	NN	O	I-protein
pRb	NN	O	I-protein
,	NN	O	O
the	NN	O	O
G1	NN	O	B-protein
cyclins	NN	O	I-protein
,	NN	O	O
the	NN	O	O
cyclin-dependent	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
and	NN	O	O
their	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

These	NN	O	O
regulators	NN	O	O
are	NN	O	O
targeted	NN	O	O
by	NN	O	O
negative	NN	O	O
growth	NN	O	O
regulatory	NN	O	O
signals	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
that	NN	O	O
provided	NN	O	O
by	NN	O	O
TGF-beta	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
either	NN	O	O
wild-type	NN	O	O
EBV	NN	O	O
or	NN	O	O
its	NN	O	O
transforming	NN	O	B-protein
latent	NN	O	I-protein
membrane	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
LMP-1	NN	O	B-protein
)	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
TGF-beta	NN	O	B-protein
1	NN	O	I-protein
-mediated	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Chemical	NN	O	O
cross-linking	NN	O	O
with	NN	O	O
125I-labeled	NN	O	B-protein
TGF-beta	NN	O	I-protein
1	NN	O	I-protein
showed	NN	O	O
an	NN	O	O
essentially	NN	O	O
normal	NN	O	O
TGF-beta	NN	O	B-protein
receptor	NN	O	I-protein
profile	NN	O	O
in	NN	O	O
EBV-positive	NN	O	B-cell_line
and	NN	O	I-cell_line
EBV-negative	NN	O	I-cell_line
Burkitt	NN	O	I-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
these	NN	O	O
receptors	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
functional	NN	O	O
in	NN	O	O
transducing	NN	O	O
signals	NN	O	O
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
the	NN	O	O
TGF-beta	NN	O	B-protein
1	NN	O	I-protein
-mediated	NN	O	O
modulation	NN	O	O
of	NN	O	O
junB	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
TGF-beta	NN	O	B-protein
1	NN	O	I-protein
did	NN	O	O
not	NN	O	O
induce	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
pRb	NN	O	B-protein
in	NN	O	O
EBV	NN	O	B-cell_line
(	NN	O	I-cell_line
or	NN	O	I-cell_line
LMP-1	NN	O	I-cell_line
)	NN	O	I-cell_line
-positive	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
opposed	NN	O	O
to	NN	O	O
EBV-negative	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
dichotomy	NN	O	O
in	NN	O	O
the	NN	O	O
TGF-beta	NN	O	B-protein
1	NN	O	I-protein
signaling	NN	O	O
pathway	NN	O	O
leading	NN	O	O
to	NN	O	O
separable	NN	O	O
gene	NN	O	O
regulatory	NN	O	O
and	NN	O	O
growth	NN	O	O
inhibitory	NN	O	O
responses	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
LMP-1	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cyclin	NN	O	B-protein
D2	NN	O	I-protein
;	NN	O	O
normal	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
EBV-negative	NN	O	B-cell_line
Burkitt	NN	O	I-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
do	NN	O	O
not	NN	O	O
express	NN	O	O
D-type	NN	O	B-protein
cyclins	NN	O	I-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
point	NN	O	O
to	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
underlying	NN	O	O
EBV-mediated	NN	O	O
B	NN	O	O
cell	NN	O	O
transformation	NN	O	O
whereby	NN	O	O
constitutive	NN	O	O
induction	NN	O	O
of	NN	O	O
key	NN	O	B-protein
cell	NN	O	I-protein
cycle	NN	O	I-protein
regulators	NN	O	I-protein
by	NN	O	O
LMP-1	NN	O	B-protein
can	NN	O	O
lead	NN	O	O
to	NN	O	O
pRb	NN	O	B-protein
hyperphosphorylation	NN	O	O
and	NN	O	O
uncontrolled	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Does	NN	O	O
thyroidectomy	NN	O	O
,	NN	O	O
radioactive	NN	O	O
iodine	NN	O	O
therapy	NN	O	O
,	NN	O	O
or	NN	O	O
antithyroid	NN	O	O
drug	NN	O	O
treatment	NN	O	O
alter	NN	O	O
reactivity	NN	O	O
of	NN	O	O
patients	NN	O	O
'	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
epitopes	NN	O	O
of	NN	O	O
thyrotropin	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
autoimmune	NN	O	O
thyroid	NN	O	O
diseases	NN	O	O
?	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
treatment	NN	O	O
on	NN	O	O
thyroid	NN	O	O
antibody	NN	O	O
production	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
reactivity	NN	O	O
to	NN	O	O
thyroid	NN	O	O
antigens	NN	O	O
was	NN	O	O
studied	NN	O	O
in	NN	O	O
15	NN	O	O
patients	NN	O	O
with	NN	O	O
Graves	NN	O	O
'	NN	O	O
disease	NN	O	O
(	NN	O	O
GD	NN	O	O
)	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
thyroidectomy	NN	O	O
,	NN	O	O
19	NN	O	O
patients	NN	O	O
with	NN	O	O
GD	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
radioactive	NN	O	O
iodine	NN	O	O
(	NN	O	O
RAI	NN	O	O
)	NN	O	O
therapy	NN	O	O
,	NN	O	O
and	NN	O	O
9	NN	O	O
patients	NN	O	O
maintained	NN	O	O
euthyroid	NN	O	O
on	NN	O	O
antithyroid	NN	O	O
drugs	NN	O	O
(	NN	O	O
ATD	NN	O	O
)	NN	O	O
.	NN	O	O

Twenty	NN	O	O
subjects	NN	O	O
matched	NN	O	O
for	NN	O	O
age	NN	O	O
and	NN	O	O
sex	NN	O	O
without	NN	O	O
known	NN	O	O
thyroid	NN	O	O
disease	NN	O	O
served	NN	O	O
as	NN	O	O
controls	NN	O	O
.	NN	O	O

In	NN	O	O
GD	NN	O	O
patients	NN	O	O
,	NN	O	O
the	NN	O	O
responses	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
TSH	NN	O	B-cell_line
receptor	NN	O	I-cell_line
(	NN	O	I-cell_line
TSHR	NN	O	I-cell_line
)	NN	O	I-cell_line
-specific	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
to	NN	O	O
recombinant	NN	O	B-protein
human	NN	O	I-protein
TSHR	NN	O	I-protein
extracellular	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
thyroglobulin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TSHR	NN	O	B-protein
peptides	NN	O	O
were	NN	O	O
examined	NN	O	O
on	NN	O	O
the	NN	O	O
day	NN	O	O
of	NN	O	O
surgery	NN	O	O
or	NN	O	O
RAI	NN	O	O
therapy	NN	O	O
(	NN	O	O
day	NN	O	O
0	NN	O	O
)	NN	O	O
and	NN	O	O
also	NN	O	O
6-8	NN	O	O
weeks	NN	O	O
and	NN	O	O
3-6	NN	O	O
months	NN	O	O
thereafter	NN	O	O
.	NN	O	O

Reactivity	NN	O	O
to	NN	O	O
TSHR	NN	O	B-protein
peptides	NN	O	O
before	NN	O	O
surgery	NN	O	O
was	NN	O	O
heterogeneous	NN	O	O
and	NN	O	O
spanned	NN	O	O
the	NN	O	O
entire	NN	O	O
extracellular	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

Six	NN	O	O
to	NN	O	O
8	NN	O	O
weeks	NN	O	O
after	NN	O	O
subtotal	NN	O	O
thyroidectomy	NN	O	O
,	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
patients	NN	O	O
'	NN	O	O
PBMC	NN	O	B-cell_type
responding	NN	O	O
to	NN	O	O
any	NN	O	O
peptide	NN	O	O
and	NN	O	O
the	NN	O	O
average	NN	O	O
number	NN	O	O
of	NN	O	O
recognized	NN	O	O
peptides	NN	O	O
decreased	NN	O	O
.	NN	O	O

A	NN	O	O
further	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
reactivity	NN	O	O
to	NN	O	O
TSHR	NN	O	B-protein
peptides	NN	O	O
was	NN	O	O
observed	NN	O	O
3-6	NN	O	O
months	NN	O	O
after	NN	O	O
surgery	NN	O	O
.	NN	O	O

The	NN	O	O
responses	NN	O	O
of	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
Graves	NN	O	O
'	NN	O	O
patients	NN	O	O
before	NN	O	O
RAI	NN	O	O
therapy	NN	O	O
were	NN	O	O
less	NN	O	O
than	NN	O	O
those	NN	O	O
in	NN	O	O
the	NN	O	O
presurgical	NN	O	O
group	NN	O	O
.	NN	O	O

Six	NN	O	O
to	NN	O	O
8	NN	O	O
weeks	NN	O	O
after	NN	O	O
RAI	NN	O	O
therapy	NN	O	O
,	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
patients	NN	O	O
responding	NN	O	O
to	NN	O	O
any	NN	O	O
peptide	NN	O	O
and	NN	O	O
the	NN	O	O
average	NN	O	O
number	NN	O	O
of	NN	O	O
recognized	NN	O	O
peptides	NN	O	O
increased	NN	O	O
.	NN	O	O

Three	NN	O	O
to	NN	O	O
6	NN	O	O
months	NN	O	O
after	NN	O	O
RAI	NN	O	O
,	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
to	NN	O	O
TSHR	NN	O	B-protein
peptides	NN	O	O
were	NN	O	O
less	NN	O	O
than	NN	O	O
those	NN	O	O
6-8	NN	O	O
weeks	NN	O	O
after	NN	O	O
RAI	NN	O	O
therapy	NN	O	O
,	NN	O	O
but	NN	O	O
still	NN	O	O
higher	NN	O	O
than	NN	O	O
the	NN	O	O
values	NN	O	O
on	NN	O	O
day	NN	O	O
0	NN	O	O
.	NN	O	O

Responses	NN	O	O
of	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
GD	NN	O	O
,	NN	O	O
maintained	NN	O	O
euthyroid	NN	O	O
on	NN	O	O
ATD	NN	O	O
,	NN	O	O
were	NN	O	O
lower	NN	O	O
than	NN	O	O
those	NN	O	O
before	NN	O	O
surgery	NN	O	O
or	NN	O	O
RAI	NN	O	O
therapy	NN	O	O
.	NN	O	O

The	NN	O	O
reactivity	NN	O	O
of	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
in	NN	O	O
different	NN	O	O
groups	NN	O	O
reflected	NN	O	O
a	NN	O	O
pattern	NN	O	O
similar	NN	O	O
to	NN	O	O
PBMC	NN	O	B-cell_type
after	NN	O	O
treatment	NN	O	O
.	NN	O	O

TSHR	NN	O	B-protein
antibody	NN	O	I-protein
and	NN	O	O
microsomal	NN	O	B-protein
antibody	NN	O	I-protein
levels	NN	O	O
decreased	NN	O	O
after	NN	O	O
surgery	NN	O	O
,	NN	O	O
but	NN	O	O
increased	NN	O	O
after	NN	O	O
RAI	NN	O	O
therapy	NN	O	O
.	NN	O	O

The	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
recognized	NN	O	O
peptides	NN	O	O
by	NN	O	O
patients	NN	O	O
'	NN	O	O
PBMC	NN	O	B-cell_type
before	NN	O	O
RAI	NN	O	O
and	NN	O	O
surgery	NN	O	O
may	NN	O	O
reflect	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
long	NN	O	O
term	NN	O	O
therapy	NN	O	O
with	NN	O	O
ATD	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
before	NN	O	O
RAI	NN	O	O
vs.	NN	O	O
the	NN	O	O
shorter	NN	O	O
period	NN	O	O
in	NN	O	O
patients	NN	O	O
before	NN	O	O
surgery	NN	O	O
.	NN	O	O

The	NN	O	O
decreased	NN	O	O
T	NN	O	O
cell	NN	O	O
reactivity	NN	O	O
to	NN	O	O
thyroid	NN	O	B-protein
antigens	NN	O	I-protein
after	NN	O	O
thyroidectomy	NN	O	O
could	NN	O	O
be	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
removal	NN	O	O
of	NN	O	O
a	NN	O	O
major	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
thyroid	NN	O	O
gland	NN	O	O
or	NN	O	O
redistribution	NN	O	O
of	NN	O	O
suppressor-inducer	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
increased	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
after	NN	O	O
RAI	NN	O	O
therapy	NN	O	O
is	NN	O	O
probably	NN	O	O
epitope	NN	O	O
specific	NN	O	O
,	NN	O	O
rather	NN	O	O
than	NN	O	O
a	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
whole	NN	O	O
TSHR	NN	O	B-protein
molecule	NN	O	O
.	NN	O	O

Synchronous	NN	O	O
recognition	NN	O	O
of	NN	O	O
peptides	NN	O	O
158-176	NN	O	O
and	NN	O	O
248-263	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
GD	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
recognition	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
epitopes	NN	O	B-protein
may	NN	O	O
be	NN	O	O
an	NN	O	O
early	NN	O	O
sign	NN	O	O
of	NN	O	O
immune	NN	O	O
remission	NN	O	O
and	NN	O	O
a	NN	O	O
predictor	NN	O	O
of	NN	O	O
euthyroidism	NN	O	O
.	NN	O	O

-DOCSTART-	O

Circumvention	NN	O	O
of	NN	O	O
tolerance	NN	O	O
for	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
T	NN	O	I-protein
cell	NN	O	I-protein
protein	NN	O	I-protein
TCF-1	NN	O	B-protein
by	NN	O	O
immunization	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
knock-out	NN	O	O
mice	NN	O	O
.	NN	O	O

Molecular	NN	O	O
events	NN	O	O
that	NN	O	O
underlie	NN	O	O
the	NN	O	O
well-defined	NN	O	O
phenotypic	NN	O	O
changes	NN	O	O
of	NN	O	O
the	NN	O	O
differentiating	NN	O	B-cell_type
thymocyte	NN	O	I-cell_type
are	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

A	NN	O	O
candidate	NN	O	O
gene	NN	O	O
to	NN	O	O
control	NN	O	O
thymocyte	NN	O	B-cell_type
differentiation	NN	O	O
,	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
factor-1	NN	O	I-protein
(	NN	O	O
TCF-1	NN	O	B-protein
)	NN	O	O
*	NN	O	O
encodes	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Its	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
pattern	NN	O	O
is	NN	O	O
complex	NN	O	O
during	NN	O	O
embryogenesis	NN	O	O
,	NN	O	O
yet	NN	O	O
restricted	NN	O	O
to	NN	O	O
lymphocytes	NN	O	B-cell_type
postnatally	NN	O	O
.	NN	O	O

Expression	NN	O	O
studies	NN	O	O
on	NN	O	O
TCF-1	NN	O	B-protein
protein	NN	O	I-protein
have	NN	O	O
been	NN	O	O
hampered	NN	O	O
by	NN	O	O
the	NN	O	O
difficulty	NN	O	O
to	NN	O	O
raise	NN	O	O
antibodies	NN	O	B-protein
due	NN	O	O
to	NN	O	O
extreme	NN	O	O
evolutionary	NN	O	O
conservation	NN	O	O
.	NN	O	O

TCF-1	NN	O	B-protein
knock-out	NN	O	O
mice	NN	O	O
,	NN	O	O
generated	NN	O	O
recently	NN	O	O
in	NN	O	O
our	NN	O	O
laboratory	NN	O	O
,	NN	O	O
have	NN	O	O
strongly	NN	O	O
decreased	NN	O	O
numbers	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
are	NN	O	O
otherwise	NN	O	O
normal	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
these	NN	O	O
mice	NN	O	O
to	NN	O	O
generate	NN	O	O
anti-TCF-1	NN	O	B-protein
antibodies	NN	O	I-protein
.	NN	O	O

By	NN	O	O
immunization	NN	O	O
with	NN	O	O
a	NN	O	O
recombinant	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
TCF-1	NN	O	B-protein
knock-out	NN	O	O
mice	NN	O	O
readily	NN	O	O
yield	NN	O	O
antiserum	NN	O	O
titers	NN	O	O
against	NN	O	O
human	NN	O	O
and	NN	O	O
mouse	NN	O	O
TCF-1	NN	O	O
protein	NN	O	O
.	NN	O	O

Wild-type	NN	O	O
littermates	NN	O	O
remain	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
TCF-1	NN	O	B-protein
while	NN	O	O
they	NN	O	O
mount	NN	O	O
a	NN	O	O
high-titer	NN	O	O
antibody	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
fusion	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
Maltose	NN	O	B-protein
Binding	NN	O	I-protein
Protein	NN	O	I-protein
(	NN	O	O
MBP	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Subsequently	NN	O	O
,	NN	O	O
TCF-1-specific	NN	O	B-cell_line
hybridomas	NN	O	I-cell_line
could	NN	O	O
be	NN	O	O
prepared	NN	O	O
from	NN	O	O
the	NN	O	O
spleens	NN	O	O
of	NN	O	O
immunized	NN	O	O
knock-out	NN	O	O
mice	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
illustrates	NN	O	O
the	NN	O	O
almost	NN	O	O
complete	NN	O	O
tolerance	NN	O	O
of	NN	O	O
mice	NN	O	O
for	NN	O	O
human	NN	O	B-protein
TCF-1	NN	O	I-protein
and	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
this	NN	O	O
tolerance	NN	O	O
is	NN	O	O
readily	NN	O	O
broken	NN	O	O
by	NN	O	O
gene	NN	O	O
knock-out	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
usefulness	NN	O	O
of	NN	O	O
knock-out	NN	O	O
mice	NN	O	O
for	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
against	NN	O	O
the	NN	O	O
gene	NN	O	O
product	NN	O	O
of	NN	O	O
interest	NN	O	O
is	NN	O	O
underscored	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
Nm23H2	NN	O	B-protein
,	NN	O	O
binds	NN	O	O
to	NN	O	O
and	NN	O	O
activates	NN	O	O
the	NN	O	O
translocated	NN	O	O
c-myc	NN	O	B-DNA
allele	NN	O	I-DNA
in	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
an	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprint	NN	O	O
over	NN	O	O
the	NN	O	O
PuF	NN	O	B-DNA
site	NN	O	I-DNA
on	NN	O	O
the	NN	O	O
translocated	NN	O	O
c-myc	NN	O	B-DNA
allele	NN	O	I-DNA
in	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
PuF	NN	O	B-DNA
site	NN	O	I-DNA
on	NN	O	O
the	NN	O	O
silent	NN	O	B-DNA
normal	NN	O	I-DNA
c-myc	NN	O	I-DNA
allele	NN	O	I-DNA
was	NN	O	O
unoccupied	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
with	NN	O	O
antibody	NN	O	O
,	NN	O	O
UV	NN	O	O
cross-linking	NN	O	O
followed	NN	O	O
by	NN	O	O
SDS-gel	NN	O	O
electrophoresis	NN	O	O
,	NN	O	O
and	NN	O	O
Western	NN	O	O
analysis	NN	O	O
that	NN	O	O
Nm23H2	NN	O	B-protein
in	NN	O	O
B	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
c-myc	NN	O	O
PuF	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

Transfection	NN	O	O
experiments	NN	O	O
with	NN	O	O
c-myc	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
both	NN	O	O
DHL-9	NN	O	B-cell_line
and	NN	O	O
Raji	NN	O	B-cell_line
cells	NN	O	I-cell_line
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
PuF	NN	O	B-DNA
site	NN	O	I-DNA
functioned	NN	O	O
as	NN	O	O
a	NN	O	O
positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
a	NN	O	O
drop	NN	O	O
in	NN	O	O
activity	NN	O	O
with	NN	O	O
mutation	NN	O	O
of	NN	O	O
this	NN	O	O
site	NN	O	O
.	NN	O	O

Access	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
is	NN	O	O
blocked	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	B-DNA
silent	NN	O	I-DNA
c-myc	NN	O	I-DNA
allele	NN	O	I-DNA
;	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
Nm23H2	NN	O	B-protein
protein	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
deregulation	NN	O	O
of	NN	O	O
the	NN	O	O
translocated	NN	O	O
c-myc	NN	O	B-DNA
allele	NN	O	I-DNA
in	NN	O	O
Burkitt	NN	O	B-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
two	NN	O	O
novel	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
5'-untranslated	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
A	NN	O	I-DNA
gamma-globin	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Interaction	NN	O	O
between	NN	O	O
the	NN	O	O
stage	NN	O	B-DNA
selector	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
SSE	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
gamma-globin	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
hypersensitivity	NN	O	B-DNA
site	NN	O	I-DNA
2	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
locus	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
partly	NN	O	O
mediates	NN	O	O
the	NN	O	O
competitive	NN	O	O
silencing	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
fetal	NN	O	O
developmental	NN	O	O
stage	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
a	NN	O	O
second	NN	O	O
SSE-like	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
5'-untranslated	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
gamma-gene	NN	O	B-DNA
also	NN	O	O
contributes	NN	O	O
to	NN	O	O
this	NN	O	O
competitive	NN	O	O
silencing	NN	O	O
of	NN	O	O
the	NN	O	O
beta-gene	NN	O	B-DNA
.	NN	O	O

Utilizing	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
in	NN	O	O
the	NN	O	O
fetal	NN	O	B-cell_line
erythroid	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
K562	NN	O	B-cell_line
,	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
enhancer	NN	O	I-DNA
of	NN	O	O
hypersensitivity	NN	O	B-DNA
site	NN	O	I-DNA
2	NN	O	I-DNA
can	NN	O	O
preferentially	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
gamma-promoter	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
SSE	NN	O	B-DNA
,	NN	O	O
completely	NN	O	O
silencing	NN	O	O
a	NN	O	O
linked	NN	O	B-DNA
beta-promoter	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
a	NN	O	O
20-base	NN	O	O
pair	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
gamma-gene	NN	O	B-DNA
5'-untranslated	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
UTR	NN	O	B-DNA
)	NN	O	O
led	NN	O	O
to	NN	O	O
derepression	NN	O	O
of	NN	O	O
beta-promoter	NN	O	B-DNA
activity	NN	O	O
.	NN	O	O

A	NN	O	O
marked	NN	O	O
activation	NN	O	O
of	NN	O	O
gamma-promoter	NN	O	B-DNA
activity	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
with	NN	O	O
this	NN	O	O
mutation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
repressor	NN	O	O
.	NN	O	O

Fine	NN	O	O
mutagenesis	NN	O	O
dissected	NN	O	O
these	NN	O	O
activities	NN	O	O
to	NN	O	O
different	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
5'-UTR	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
stage	NN	O	O
selector	NN	O	O
activity	NN	O	O
was	NN	O	O
localized	NN	O	O
to	NN	O	O
a	NN	O	O
region	NN	O	O
centered	NN	O	O
on	NN	O	O
nucleotides	NN	O	B-DNA
+13	NN	O	I-DNA
to	NN	O	I-DNA
+15	NN	O	I-DNA
.	NN	O	O

Electromobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
utilizing	NN	O	O
this	NN	O	O
sequence	NN	O	O
demonstrated	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
fetal	NN	O	B-protein
and	NN	O	I-protein
erythroid-specific	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
repressor	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
5'-UTR	NN	O	B-DNA
was	NN	O	O
localized	NN	O	O
to	NN	O	O
tandem	NN	O	O
GATA-like	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
appear	NN	O	O
to	NN	O	O
bind	NN	O	O
a	NN	O	O
complex	NN	O	O
of	NN	O	O
two	NN	O	O
proteins	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
the	NN	O	O
erythroid	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
5'-UTR	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
gamma-gene	NN	O	B-DNA
contains	NN	O	O
sequences	NN	O	O
that	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
its	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
developmental	NN	O	O
regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Coupling	NN	O	O
of	NN	O	O
a	NN	O	O
signal	NN	O	B-protein
response	NN	O	I-protein
domain	NN	O	I-protein
in	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
to	NN	O	O
multiple	NN	O	O
pathways	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

The	NN	O	O
eukaryotic	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
and	NN	O	O
in	NN	O	O
many	NN	O	O
aspects	NN	O	O
of	NN	O	O
the	NN	O	O
genetic	NN	O	O
program	NN	O	O
mediating	NN	O	O
normal	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
and	NN	O	O
growth	NN	O	O
.	NN	O	O

The	NN	O	O
nuclear	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
tightly	NN	O	O
regulated	NN	O	O
from	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
compartment	NN	O	O
by	NN	O	O
an	NN	O	O
inhibitory	NN	O	B-protein
subunit	NN	O	I-protein
called	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

This	NN	O	O
cytoplasmic	NN	O	O
inhibitor	NN	O	O
is	NN	O	O
rapidly	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
degraded	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
diverse	NN	O	O
set	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B-inducing	NN	O	I-protein
agents	NN	O	I-protein
,	NN	O	O
including	NN	O	O
T-cell	NN	O	B-protein
mitogens	NN	O	I-protein
,	NN	O	I-protein
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
and	NN	O	O
viral	NN	O	B-protein
transactivators	NN	O	I-protein
such	NN	O	O
as	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
.	NN	O	O

To	NN	O	O
explore	NN	O	O
these	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
-dependent	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
induction	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
novel	NN	O	O
mutants	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
that	NN	O	O
uncouple	NN	O	O
its	NN	O	O
inhibitory	NN	O	O
and	NN	O	O
signal-transducing	NN	O	O
functions	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Specifically	NN	O	O
,	NN	O	O
removal	NN	O	O
of	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
36	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
failed	NN	O	O
to	NN	O	O
disrupt	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
form	NN	O	O
latent	NN	O	O
complexes	NN	O	O
with	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
deletion	NN	O	O
mutation	NN	O	O
prevented	NN	O	O
the	NN	O	O
induced	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
degradative	NN	O	O
loss	NN	O	O
,	NN	O	O
and	NN	O	O
functional	NN	O	O
release	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
from	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
Tax-expressing	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Alanine	NN	O	O
substitutions	NN	O	O
introduced	NN	O	O
at	NN	O	O
two	NN	O	O
serine	NN	O	O
residues	NN	O	O
positioned	NN	O	O
within	NN	O	O
this	NN	O	O
N-terminal	NN	O	B-protein
regulatory	NN	O	I-protein
region	NN	O	I-protein
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
also	NN	O	O
yielded	NN	O	O
constitutive	NN	O	O
repressors	NN	O	O
that	NN	O	O
escaped	NN	O	O
from	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
turnover	NN	O	O
and	NN	O	O
that	NN	O	O
potently	NN	O	O
inhibited	NN	O	O
immune	NN	O	O
activation	NN	O	O
pathways	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
induction	NN	O	O
,	NN	O	O
including	NN	O	O
those	NN	O	O
initiated	NN	O	O
from	NN	O	O
antigen	NN	O	B-protein
and	NN	O	I-protein
cytokine	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
introduction	NN	O	O
of	NN	O	O
a	NN	O	O
phosphoserine	NN	O	O
mimetic	NN	O	O
at	NN	O	O
these	NN	O	O
sites	NN	O	O
rectified	NN	O	O
this	NN	O	O
functional	NN	O	O
defect	NN	O	O
,	NN	O	O
a	NN	O	O
finding	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
causal	NN	O	O
linkage	NN	O	O
between	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
status	NN	O	O
and	NN	O	O
proteolytic	NN	O	O
stability	NN	O	O
of	NN	O	O
this	NN	O	O
cytoplasmic	NN	O	B-protein
inhibitor	NN	O	I-protein
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
in	NN	O	O
vivo	NN	O	O
studies	NN	O	O
define	NN	O	O
a	NN	O	O
critical	NN	O	O
signal	NN	O	O
response	NN	O	O
domain	NN	O	O
in	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
that	NN	O	O
coordinately	NN	O	O
controls	NN	O	O
the	NN	O	O
biologic	NN	O	O
activities	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
viral	NN	O	O
and	NN	O	O
immune	NN	O	O
stimuli	NN	O	O
.	NN	O	O

-DOCSTART-	O

Growth	NN	O	O
regulation	NN	O	O
and	NN	O	O
cellular	NN	O	O
changes	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
prostatic	NN	O	I-cell_line
cancer	NN	O	I-cell_line
LNCaP	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
induced	NN	O	O
by	NN	O	O
T	NN	O	O
lymphocyte-conditioned	NN	O	O
medium	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_line
prostatic	NN	O	I-cell_line
epithelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
an	NN	O	O
androgen-dependent	NN	O	B-cell_line
LNCaP	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
were	NN	O	O
examined	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
conditioned	NN	O	O
medium	NN	O	O
(	NN	O	O
CM	NN	O	O
)	NN	O	O
derived	NN	O	O
from	NN	O	O
phytohemagglutinin	NN	O	B-cell_line
(	NN	O	I-cell_line
PHA	NN	O	I-cell_line
)	NN	O	I-cell_line
-stimulated	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
CM	NN	O	O
caused	NN	O	O
a	NN	O	O
greater	NN	O	O
than	NN	O	O
70	NN	O	O
%	NN	O	O
reduction	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
by	NN	O	O
cell	NN	O	O
counting	NN	O	O
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

These	NN	O	O
cells	NN	O	O
showed	NN	O	O
G1	NN	O	O
phase	NN	O	O
arrest	NN	O	O
and	NN	O	O
the	NN	O	O
clonogenicity	NN	O	O
was	NN	O	O
reduced	NN	O	O
.	NN	O	O

The	NN	O	O
growth-modulating	NN	O	O
effect	NN	O	O
was	NN	O	O
dose-dependent	NN	O	O
and	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
cell	NN	O	O
lysis	NN	O	O
or	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
androgen	NN	O	O
to	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
on	NN	O	O
these	NN	O	O
cells	NN	O	O
showed	NN	O	O
approximately	NN	O	O
50	NN	O	O
%	NN	O	O
reduction	NN	O	O
,	NN	O	O
underlining	NN	O	O
a	NN	O	O
proliferation	NN	O	O
reduction	NN	O	O
mechanism	NN	O	O
.	NN	O	O

The	NN	O	O
prostate-specific	NN	O	B-protein
antigen	NN	O	I-protein
(	NN	O	O
PSA	NN	O	B-protein
)	NN	O	O
was	NN	O	O
downregulated	NN	O	O
to	NN	O	O
approximately	NN	O	O
75	NN	O	O
%	NN	O	O
during	NN	O	O
the	NN	O	O
process	NN	O	O
.	NN	O	O

Cell	NN	O	O
morphology	NN	O	O
showed	NN	O	O
dendritic	NN	O	O
processes	NN	O	O
extending	NN	O	O
from	NN	O	O
cytoplasm	NN	O	O
and	NN	O	O
other	NN	O	O
neuroendocrine	NN	O	B-cell_type
cell	NN	O	I-cell_type
characteristics	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
cytoskeleton	NN	O	B-protein
and	NN	O	I-protein
intracellular	NN	O	I-protein
proteins	NN	O	I-protein
increased	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
immunostaining	NN	O	O
on	NN	O	O
slides	NN	O	O
and	NN	O	O
by	NN	O	O
ELISA	NN	O	O
procedures	NN	O	O
.	NN	O	O

These	NN	O	O
included	NN	O	O
vimentin	NN	O	B-protein
,	NN	O	O
correlating	NN	O	O
to	NN	O	O
cell	NN	O	O
shape	NN	O	O
changes	NN	O	O
,	NN	O	O
cytokeratins	NN	O	B-protein
8	NN	O	I-protein
and	NN	O	I-protein
18	NN	O	I-protein
,	NN	O	O
associated	NN	O	O
with	NN	O	O
differentiated	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
of	NN	O	O
prostate	NN	O	O
epithelia	NN	O	O
,	NN	O	O
and	NN	O	O
neuron-specific	NN	O	B-protein
enolase	NN	O	I-protein
and	NN	O	O
serotonin	NN	O	O
,	NN	O	O
associated	NN	O	O
with	NN	O	O
neuroendocrine	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

From	NN	O	O
these	NN	O	O
cellular	NN	O	O
changes	NN	O	O
,	NN	O	O
we	NN	O	O
can	NN	O	O
infer	NN	O	O
that	NN	O	O
the	NN	O	O
cell	NN	O	O
growth	NN	O	O
was	NN	O	O
modulated	NN	O	O
along	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
providing	NN	O	O
the	NN	O	O
modulating	NN	O	O
activity	NN	O	O
.	NN	O	O

This	NN	O	O
growth	NN	O	B-protein
modulator	NN	O	I-protein
was	NN	O	O
semipurified	NN	O	O
and	NN	O	O
had	NN	O	O
an	NN	O	O
estimated	NN	O	O
molecular	NN	O	O
weight	NN	O	O
13	NN	O	O
,	NN	O	O
000	NN	O	O
to	NN	O	O
24	NN	O	O
,	NN	O	O
000	NN	O	O
Da	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
was	NN	O	O
determined	NN	O	O
to	NN	O	O
be	NN	O	O
distinct	NN	O	O
from	NN	O	O
TGF	NN	O	B-protein
,	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
and	NN	O	O
some	NN	O	O
commonly	NN	O	O
known	NN	O	O
lymphokines	NN	O	B-protein
.	NN	O	O

The	NN	O	O
interaction	NN	O	O
between	NN	O	O
lymphoid	NN	O	B-cell_type
and	NN	O	I-cell_type
prostatic	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
growth	NN	O	O
and	NN	O	O
development	NN	O	O
is	NN	O	O
described	NN	O	O
.	NN	O	O

-DOCSTART-	O

Platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
stimulates	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
heparin-binding	NN	O	B-protein
epidermal	NN	O	I-protein
growth	NN	O	I-protein
factor-like	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Correlation	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
kappa	NN	O	O
B	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Human	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
monocytes	NN	O	B-cell_type
responded	NN	O	O
to	NN	O	O
stimulation	NN	O	O
of	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
PAF	NN	O	B-protein
)	NN	O	O
with	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
transcript	NN	O	O
for	NN	O	O
heparin-binding	NN	O	B-protein
epidermal	NN	O	I-protein
growth	NN	O	I-protein
factor-like	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
HB-EGF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	B-protein
mitogen	NN	O	I-protein
for	NN	O	O
vascular	NN	O	B-cell_type
smooth	NN	O	I-cell_type
muscle	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
function	NN	O	O
of	NN	O	O
PAF	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
at	NN	O	O
nanomolar	NN	O	O
concentrations	NN	O	O
of	NN	O	O
the	NN	O	O
ligand	NN	O	O
,	NN	O	O
starting	NN	O	O
at	NN	O	O
30	NN	O	O
min	NN	O	O
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
PAF	NN	O	B-protein
-induced	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
HB-EGF	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
kappa	NN	O	O
B	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
functions	NN	O	O
of	NN	O	O
PAF	NN	O	B-protein
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
PAF	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
as	NN	O	O
two	NN	O	O
PAF	NN	O	B-protein
receptor	NN	O	O
antagonists	NN	O	O
,	NN	O	O
WEB	NN	O	O
2086	NN	O	O
and	NN	O	O
L-659	NN	O	O
,	NN	O	O
989	NN	O	O
,	NN	O	O
blocked	NN	O	O
both	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
HB-EGF	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
kappa	NN	O	O
B	NN	O	O
binding	NN	O	O
activity	NN	O	O
induced	NN	O	O
by	NN	O	O
PAF	NN	O	B-protein
.	NN	O	O

The	NN	O	O
antagonists	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
phorbol	NN	O	O
ester-induced	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
HB-EGF	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
kappa	NN	O	O
B	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
with	NN	O	O
pertussis	NN	O	O
toxin	NN	O	O
inhibited	NN	O	O
these	NN	O	O
functions	NN	O	O
of	NN	O	O
PAF	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
cholera	NN	O	O
toxin	NN	O	O
had	NN	O	O
no	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
.	NN	O	O

Pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
PAF	NN	O	B-protein
-stimulated	NN	O	O
kappa	NN	O	O
B	NN	O	O
binding	NN	O	O
activity	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
HB-EGF	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
PAF	NN	O	B-protein
in	NN	O	O
HB-EGF	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
this	NN	O	O
stimulation	NN	O	O
may	NN	O	O
occur	NN	O	O
through	NN	O	O
increased	NN	O	O
kappa	NN	O	O
B	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mapping	NN	O	O
of	NN	O	O
the	NN	O	O
interaction	NN	O	B-protein
site	NN	O	I-protein
of	NN	O	O
the	NN	O	O
defective	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
class	NN	O	B-cell_line
II	NN	O	I-cell_line
major	NN	O	I-cell_line
histocompatibility	NN	O	I-cell_line
complex	NN	O	I-cell_line
mutant	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
clone-13	NN	O	I-cell_line
to	NN	O	O
the	NN	O	O
divergent	NN	O	B-DNA
X2-box	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
described	NN	O	O
a	NN	O	O
mutant	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
Clone-13	NN	O	B-cell_line
,	NN	O	I-cell_line
that	NN	O	O
expresses	NN	O	O
HLA-DQ	NN	O	B-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
and	NN	O	I-protein
-DP	NN	O	I-protein
.	NN	O	O

Several	NN	O	O
criteria	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
defect	NN	O	O
in	NN	O	O
this	NN	O	O
cell	NN	O	O
line	NN	O	O
influences	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
an	NN	O	O
isotype-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
HLA-DRA	NN	O	B-DNA
and	NN	O	I-DNA
DQB	NN	O	I-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
affected	NN	O	O
factor	NN	O	O
operates	NN	O	O
via	NN	O	O
cis-elements	NN	O	B-DNA
located	NN	O	O
between	NN	O	O
-141	NN	O	B-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
series	NN	O	O
of	NN	O	O
hybrid	NN	O	B-DNA
DRA/DQB	NN	O	I-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
was	NN	O	O
generated	NN	O	O
to	NN	O	O
further	NN	O	O
map	NN	O	O
the	NN	O	O
relevant	NN	O	O
cis-elements	NN	O	B-DNA
in	NN	O	O
this	NN	O	O
system	NN	O	O
.	NN	O	O

Insertion	NN	O	O
of	NN	O	O
oligonucleotides	NN	O	O
spanning	NN	O	O
the	NN	O	O
DQB	NN	O	B-DNA
X-box	NN	O	I-DNA
(	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
DQB-W	NN	O	B-DNA
region	NN	O	I-DNA
or	NN	O	O
the	NN	O	O
DQB	NN	O	B-DNA
Y-box	NN	O	I-DNA
)	NN	O	O
upstream	NN	O	B-DNA
of	NN	O	I-DNA
-141	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
DRA	NN	O	B-DNA
reporter	NN	O	I-DNA
plasmid	NN	O	I-DNA
rescued	NN	O	O
expression	NN	O	O
to	NN	O	O
nearly	NN	O	O
wild-type	NN	O	O
levels	NN	O	O
.	NN	O	O

Substitution	NN	O	O
promoters	NN	O	O
were	NN	O	O
then	NN	O	O
generated	NN	O	O
where	NN	O	O
the	NN	O	O
entire	NN	O	O
X-box	NN	O	B-DNA
,	NN	O	O
or	NN	O	O
only	NN	O	O
the	NN	O	O
X1-	NN	O	B-DNA
or	NN	O	I-DNA
X2-boxes	NN	O	I-DNA
of	NN	O	O
HLA-DRA	NN	O	B-DNA
were	NN	O	O
replaced	NN	O	O
with	NN	O	O
the	NN	O	O
analogous	NN	O	O
regions	NN	O	O
of	NN	O	O
HLA-DQB	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
DQB	NN	O	B-DNA
X2-box	NN	O	I-DNA
was	NN	O	O
able	NN	O	O
to	NN	O	O
restore	NN	O	O
expression	NN	O	O
to	NN	O	O
the	NN	O	O
silent	NN	O	B-DNA
DRA	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
replacement	NN	O	O
of	NN	O	O
the	NN	O	O
DQB	NN	O	B-DNA
X2-box	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
X2-box	NN	O	I-DNA
markedly	NN	O	O
diminished	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
DQB	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
mutant	NN	O	B-cell_line
cell	NN	O	I-cell_line
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
hybrid	NN	O	B-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
were	NN	O	O
defective	NN	O	O
when	NN	O	O
transfected	NN	O	O
into	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
,	NN	O	O
HLA-DR/-DQ	NN	O	B-cell_line
positive	NN	O	I-cell_line
parental	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
Jijoye	NN	O	B-cell_line
.	NN	O	O

These	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
divergent	NN	O	B-DNA
X2-box	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
major	NN	O	I-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
promoters	NN	O	I-DNA
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
influencing	NN	O	O
differential	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
isotypes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Restoration	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
ZEBRA	NN	O	I-protein
protein	NN	O	I-protein
's	NN	O	O
capacity	NN	O	O
to	NN	O	O
disrupt	NN	O	O
latency	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
heterologous	NN	O	B-protein
activation	NN	O	I-protein
regions	NN	O	I-protein
.	NN	O	O

The	NN	O	O
ZEBRA	NN	O	B-protein
protein	NN	O	I-protein
has	NN	O	O
a	NN	O	O
unique	NN	O	O
biological	NN	O	O
function	NN	O	O
among	NN	O	O
herpesviral	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
disruption	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
latency	NN	O	O
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
.	NN	O	O

ZEBRA	NN	O	B-protein
is	NN	O	O
a	NN	O	O
bZIP	NN	O	B-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
which	NN	O	O
binds	NN	O	O
as	NN	O	O
a	NN	O	O
dimer	NN	O	O
to	NN	O	O
7-bp	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
within	NN	O	O
EBV	NN	O	B-DNA
promoters	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
directly	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
virus	NN	O	O
replication	NN	O	O
at	NN	O	O
the	NN	O	O
EBV	NN	O	B-DNA
lytic	NN	O	I-DNA
origin	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
employed	NN	O	O
the	NN	O	O
ZEBRA	NN	O	B-protein
/EBV	NN	O	O
biological	NN	O	O
system	NN	O	O
to	NN	O	O
test	NN	O	O
whether	NN	O	O
a	NN	O	O
heterologous	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
can	NN	O	O
substitute	NN	O	O
for	NN	O	O
another	NN	O	O
activation	NN	O	O
domain	NN	O	O
(	NN	O	O
the	NN	O	O
ZEBRA	NN	O	B-protein
domain	NN	O	I-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
ZEBRA	NN	O	B-protein
activation	NN	O	I-protein
region	NN	O	I-protein
was	NN	O	O
replaced	NN	O	O
with	NN	O	O
the	NN	O	O
potent	NN	O	O
acid	NN	O	B-protein
activation	NN	O	I-protein
region	NN	O	I-protein
from	NN	O	O
the	NN	O	O
herpes	NN	O	B-protein
simplex	NN	O	I-protein
virus	NN	O	I-protein
VP16	NN	O	I-protein
protein	NN	O	I-protein
or	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
R	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Both	NN	O	O
chimeras	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
transactivate	NN	O	O
model	NN	O	O
and	NN	O	O
native	NN	O	O
promoters	NN	O	O
at	NN	O	O
equivalent	NN	O	O
or	NN	O	O
better	NN	O	O
levels	NN	O	O
than	NN	O	O
ZEBRA	NN	O	B-protein
itself	NN	O	O
.	NN	O	O

Activation	NN	O	O
was	NN	O	O
not	NN	O	O
target-	NN	O	O
or	NN	O	O
cell-type	NN	O	O
dependent	NN	O	O
,	NN	O	O
nor	NN	O	O
was	NN	O	O
it	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
virus	NN	O	O
.	NN	O	O

These	NN	O	O
activation	NN	O	O
domains	NN	O	O
restored	NN	O	O
ZEBRA	NN	O	B-protein
's	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
early	NN	O	B-protein
antigen	NN	O	I-protein
and	NN	O	O
to	NN	O	O
stimulate	NN	O	O
origin	NN	O	O
replication	NN	O	O
to	NN	O	O
levels	NN	O	O
that	NN	O	O
were	NN	O	O
equal	NN	O	O
to	NN	O	O
or	NN	O	O
greater	NN	O	O
than	NN	O	O
those	NN	O	O
of	NN	O	O
wild	NN	O	O
type	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
specificities	NN	O	O
of	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
known	NN	O	O
biological	NN	O	O
functions	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-protein
are	NN	O	O
not	NN	O	O
dependent	NN	O	O
upon	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
activation	NN	O	B-protein
domain	NN	O	I-protein
present	NN	O	I-protein
within	NN	O	O
ZEBRA	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Interleukin	NN	O	B-protein
4	NN	O	I-protein
activates	NN	O	O
a	NN	O	O
signal	NN	O	O
transducer	NN	O	O
and	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
(	NN	O	I-protein
Stat	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
which	NN	O	O
interacts	NN	O	O
with	NN	O	O
an	NN	O	O
interferon-gamma	NN	O	B-DNA
activation	NN	O	I-DNA
site-like	NN	O	I-DNA
sequence	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
I	NN	O	B-DNA
epsilon	NN	O	I-DNA
exon	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Evidence	NN	O	O
for	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
and	NN	O	O
interleukin-4	NN	O	B-protein
Stat	NN	O	I-protein
.	NN	O	O

Germ	NN	O	B-RNA
line	NN	O	I-RNA
C	NN	O	I-RNA
transcripts	NN	O	I-RNA
can	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
BL-2	NN	O	B-cell_line
.	NN	O	O

Utilizing	NN	O	O
a	NN	O	O
IFN-gamma	NN	O	B-DNA
activation	NN	O	I-DNA
site-like	NN	O	I-DNA
DNA	NN	O	I-DNA
sequence	NN	O	I-DNA
element	NN	O	I-DNA
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
I	NN	O	B-DNA
epsilon	NN	O	I-DNA
exon	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
induced	NN	O	O
a	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
and	NN	O	O
nucleus	NN	O	O
of	NN	O	O
BL-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
factor	NN	O	O
was	NN	O	O
designated	NN	O	O
IL-4	NN	O	B-protein
NAF	NN	O	I-protein
(	NN	O	O
IL-4-induced	NN	O	B-protein
nuclear-activating	NN	O	I-protein
factors	NN	O	I-protein
)	NN	O	O
and	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
tyrosine	NN	O	B-protein
phosphoprotein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
translocates	NN	O	O
from	NN	O	O
the	NN	O	O
cytosol	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
upon	NN	O	O
IL-4	NN	O	B-protein
treatment	NN	O	O
.	NN	O	O

Because	NN	O	O
these	NN	O	O
are	NN	O	O
the	NN	O	O
characteristics	NN	O	O
of	NN	O	O
a	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
Stat	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
we	NN	O	O
determined	NN	O	O
whether	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	I-protein
Stat	NN	O	I-protein
proteins	NN	O	I-protein
will	NN	O	O
interfere	NN	O	O
with	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	I-protein
IL-4	NN	O	I-protein
Stat	NN	O	I-protein
,	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
Stat6	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	O
other	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
interfere	NN	O	O
with	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
NAF	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Congruous	NN	O	O
with	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
a	NN	O	O
Stat	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
IL-4	NN	O	B-protein
induced	NN	O	O
robust	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
JAK3	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
in	NN	O	O
BL-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
with	NN	O	O
IL-4	NN	O	B-protein
Stat	NN	O	I-protein
into	NN	O	O
COS-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
produced	NN	O	O
an	NN	O	O
intracellular	NN	O	O
activity	NN	O	O
which	NN	O	O
bound	NN	O	O
the	NN	O	O
same	NN	O	O
IFN-gamma	NN	O	B-DNA
activation	NN	O	I-DNA
site-like	NN	O	I-DNA
sequence	NN	O	I-DNA
and	NN	O	O
comigrated	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
NAF	NN	O	I-protein
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
NAF	NN	O	I-protein
is	NN	O	O
IL-4	NN	O	B-protein
Stat	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
JAK3	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
receptor	NN	O	O
engagement	NN	O	O
.	NN	O	O

-DOCSTART-	O

Does	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
TAL1	NN	O	B-DNA
gene	NN	O	I-DNA
occur	NN	O	O
in	NN	O	O
a	NN	O	O
majority	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
T-cell	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
?	NN	O	O
A	NN	O	O
pediatric	NN	O	O
oncology	NN	O	O
group	NN	O	O
study	NN	O	O
.	NN	O	O

Almost	NN	O	O
25	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
T-cell	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
T-ALL	NN	O	O
)	NN	O	O
have	NN	O	O
tumor-specific	NN	O	O
rearrangements	NN	O	O
of	NN	O	O
the	NN	O	O
TAL1	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Although	NN	O	O
TAL1	NN	O	B-protein
expression	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
TAL1	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
are	NN	O	O
readily	NN	O	O
detected	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
harbor	NN	O	O
a	NN	O	O
rearranged	NN	O	B-DNA
TAL1	NN	O	I-DNA
allele	NN	O	I-DNA
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
proposed	NN	O	O
that	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
TAL1	NN	O	B-protein
promotes	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
T-ALL	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
TAL1	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
most	NN	O	O
patients	NN	O	O
with	NN	O	O
T-ALL	NN	O	O
,	NN	O	O
including	NN	O	O
many	NN	O	O
that	NN	O	O
do	NN	O	O
not	NN	O	O
display	NN	O	O
an	NN	O	O
apparent	NN	O	O
TAL1	NN	O	B-DNA
gene	NN	O	I-DNA
alteration	NN	O	O
.	NN	O	O

A	NN	O	O
polymorphic	NN	O	O
dinucleotide	NN	O	O
repeat	NN	O	O
in	NN	O	O
the	NN	O	O
transcribed	NN	O	O
sequences	NN	O	O
of	NN	O	O
TAL1	NN	O	B-protein
was	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
allele	NN	O	O
specificity	NN	O	O
of	NN	O	O
TAL1	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
T-ALL	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Monoallelic	NN	O	O
expression	NN	O	O
of	NN	O	O
TAL1	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
all	NN	O	O
patients	NN	O	O
(	NN	O	O
8	NN	O	O
of	NN	O	O
8	NN	O	O
)	NN	O	O
bearing	NN	O	O
a	NN	O	O
TAL1	NN	O	B-DNA
gene	NN	O	I-DNA
rearrangement	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
without	NN	O	O
detectable	NN	O	O
TAL1	NN	O	B-protein
rearrangements	NN	O	O
,	NN	O	O
TAL1	NN	O	B-protein
transcription	NN	O	O
occurred	NN	O	O
in	NN	O	O
either	NN	O	O
a	NN	O	O
monoallelic	NN	O	O
(	NN	O	O
3	NN	O	O
of	NN	O	O
7	NN	O	O
patients	NN	O	O
)	NN	O	O
or	NN	O	O
a	NN	O	O
biallelic	NN	O	O
(	NN	O	O
4	NN	O	O
of	NN	O	O
7	NN	O	O
patients	NN	O	O
)	NN	O	O
fashion	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
TAL1	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
these	NN	O	O
patients	NN	O	O
may	NN	O	O
result	NN	O	O
from	NN	O	O
subtle	NN	O	O
alterations	NN	O	O
in	NN	O	O
cis-acting	NN	O	B-DNA
regulatory	NN	O	I-DNA
sequences	NN	O	I-DNA
(	NN	O	O
affecting	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	B-DNA
TAL1	NN	O	I-DNA
allele	NN	O	I-DNA
)	NN	O	O
or	NN	O	O
changes	NN	O	O
in	NN	O	O
trans-acting	NN	O	O
factors	NN	O	O
that	NN	O	O
control	NN	O	O
TAL1	NN	O	B-protein
transcription	NN	O	O
(	NN	O	O
affecting	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
TAL1	NN	O	B-DNA
alleles	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
erythroid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
acute	NN	O	O
megakaryoblastic	NN	O	O
leukaemia	NN	O	O
and	NN	O	O
transient	NN	O	O
myeloproliferative	NN	O	O
disorder	NN	O	O
in	NN	O	O
Down	NN	O	O
's	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Acute	NN	O	O
megakaryoblastic	NN	O	O
leukaemia	NN	O	O
(	NN	O	O
M7	NN	O	O
)	NN	O	O
and	NN	O	O
transient	NN	O	O
myeloproliferative	NN	O	O
disorder	NN	O	O
in	NN	O	O
Down	NN	O	O
's	NN	O	O
syndrome	NN	O	O
(	NN	O	O
TMD	NN	O	O
)	NN	O	O
are	NN	O	O
characterized	NN	O	O
by	NN	O	O
rapid	NN	O	O
growth	NN	O	O
of	NN	O	O
abnormal	NN	O	B-cell_type
blast	NN	O	I-cell_type
cells	NN	O	I-cell_type
which	NN	O	O
express	NN	O	O
megakaryocytic	NN	O	B-protein
markers	NN	O	I-protein
.	NN	O	O

To	NN	O	O
clarify	NN	O	O
properties	NN	O	O
of	NN	O	O
the	NN	O	O
blast	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
M7	NN	O	O
and	NN	O	O
TMD	NN	O	O
cases	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
erythroid	NN	O	O
markers	NN	O	O
expression	NN	O	O
in	NN	O	O
blasts	NN	O	B-cell_type
from	NN	O	O
six	NN	O	O
cases	NN	O	O
with	NN	O	O
M7	NN	O	O
and	NN	O	O
seven	NN	O	O
cases	NN	O	O
with	NN	O	O
TMD	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
.	NN	O	O

Erythroid-specific	NN	O	B-RNA
mRNAs	NN	O	I-RNA
encoding	NN	O	O
gamma-globin	NN	O	B-protein
and	NN	O	O
erythroid	NN	O	B-protein
delta-aminolevulinate	NN	O	I-protein
synthase	NN	O	I-protein
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
in	NN	O	O
blasts	NN	O	B-cell_type
from	NN	O	O
most	NN	O	O
of	NN	O	O
these	NN	O	O
cases	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
majorities	NN	O	O
of	NN	O	O
the	NN	O	O
blasts	NN	O	B-cell_type
in	NN	O	O
M7	NN	O	O
and	NN	O	O
TMD	NN	O	O
cases	NN	O	O
have	NN	O	O
erythroid	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
found	NN	O	O
that	NN	O	O
mRNAs	NN	O	B-RNA
encoding	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
GATA-2	NN	O	B-protein
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
all	NN	O	O
these	NN	O	O
cases	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
M7	NN	O	B-cell_type
blasts	NN	O	I-cell_type
and	NN	O	O
TMD	NN	O	B-cell_type
blasts	NN	O	I-cell_type
correspond	NN	O	O
to	NN	O	O
the	NN	O	O
erythroid/megakaryocytic	NN	O	B-cell_type
bipotential	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
Ah	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
TCDD	NN	O	B-protein
receptor	NN	O	I-protein
)	NN	O	O
during	NN	O	O
human	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
found	NN	O	O
a	NN	O	O
high	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-RNA
Ah	NN	O	I-RNA
receptor	NN	O	I-RNA
(	NN	O	I-RNA
TCDD	NN	O	I-RNA
receptor	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
individuals	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
in	NN	O	O
blood	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
first	NN	O	O
investigated	NN	O	O
in	NN	O	O
fractions	NN	O	O
of	NN	O	O
nucleated	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
revealing	NN	O	O
predominant	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Ah	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
monocyte	NN	O	B-cell_type
fraction	NN	O	I-cell_type
.	NN	O	O

Then	NN	O	O
the	NN	O	O
expression	NN	O	O
levels	NN	O	O
of	NN	O	O
AhR	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
various	NN	O	O
hematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
examined	NN	O	O
together	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
Arnt	NN	O	B-protein
and	NN	O	O
P450IA1	NN	O	B-protein
.	NN	O	O

AhR	NN	O	B-protein
was	NN	O	O
expressed	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
in	NN	O	O
monocytoid	NN	O	B-cell_line
U937	NN	O	I-cell_line
,	NN	O	O
THP1	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
HEL/S	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
at	NN	O	O
moderate	NN	O	O
levels	NN	O	O
in	NN	O	O
promyelocytic	NN	O	B-cell_line
HL60	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
erythroblastic	NN	O	B-cell_line
HEL	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
MOLT4	NN	O	B-cell_line
(	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
)	NN	O	O
and	NN	O	O
BALL1	NN	O	B-cell_line
(	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
)	NN	O	O
,	NN	O	O
nor	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
erythroblasts	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
a	NN	O	O
specific	NN	O	O
induction	NN	O	O
of	NN	O	O
AhR	NN	O	B-protein
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
HL60	NN	O	B-cell_line
and	NN	O	O
HEL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
toward	NN	O	O
monocytes-macrophages	NN	O	B-cell_type
by	NN	O	O
incubation	NN	O	O
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
,	NN	O	O
showing	NN	O	O
a	NN	O	O
5-	NN	O	O
to	NN	O	O
2-fold	NN	O	O
increase	NN	O	O
of	NN	O	O
AhR	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

The	NN	O	O
incubation	NN	O	O
with	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
beta	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
5-	NN	O	O
to	NN	O	O
7-fold	NN	O	O
increase	NN	O	O
of	NN	O	O
AhR	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

The	NN	O	O
HEL	NN	O	B-cell_line
cells	NN	O	I-cell_line
also	NN	O	O
exhibited	NN	O	O
a	NN	O	O
similar	NN	O	O
elevation	NN	O	O
of	NN	O	O
AhR	NN	O	B-RNA
mRNA	NN	O	I-RNA
level	NN	O	O
,	NN	O	O
when	NN	O	O
they	NN	O	O
had	NN	O	O
differentiated	NN	O	O
toward	NN	O	O
monocyte-macrophage	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
these	NN	O	O
combined	NN	O	O
inducers	NN	O	O
,	NN	O	O
but	NN	O	O
little	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
was	NN	O	O
observed	NN	O	O
when	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
into	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
the	NN	O	O
differentiated	NN	O	B-cell_line
HL60	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
3-methylcholanthrene	NN	O	O
,	NN	O	O
a	NN	O	O
ligand	NN	O	O
of	NN	O	O
AhR	NN	O	B-protein
,	NN	O	O
induced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
P450IA1	NN	O	B-protein
gene	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicated	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
AhR	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
significantly	NN	O	O
induced	NN	O	O
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
differentiated	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
responsive	NN	O	O
to	NN	O	O
xenobiotics	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
AhR	NN	O	B-protein
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
also	NN	O	O
in	NN	O	O
the	NN	O	O
eventual	NN	O	O
activation	NN	O	O
of	NN	O	O
environmental	NN	O	O
carcinogens	NN	O	O
.	NN	O	O

-DOCSTART-	O

Neutrophils	NN	O	B-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
express	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
PU.1	NN	O	B-protein
(	NN	O	O
Spi-1	NN	O	B-protein
)	NN	O	O
but	NN	O	O
not	NN	O	O
Spi-B	NN	O	B-protein
.	NN	O	O

PU.1	NN	O	B-protein
(	NN	O	O
the	NN	O	O
Spi-1	NN	O	B-DNA
oncogene	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
Spi-B	NN	O	B-protein
are	NN	O	O
closely	NN	O	O
related	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
ets	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
sharing	NN	O	O
similar	NN	O	O
DNA	NN	O	O
binding	NN	O	O
specificities	NN	O	O
mediated	NN	O	O
by	NN	O	O
similar	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domains	NN	O	I-protein
.	NN	O	O

PU.1	NN	O	B-protein
and	NN	O	O
Spi-B	NN	O	B-protein
have	NN	O	O
been	NN	O	O
previously	NN	O	O
described	NN	O	O
as	NN	O	O
being	NN	O	O
predominantly	NN	O	O
expressed	NN	O	O
coordinately	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
their	NN	O	O
expression	NN	O	O
in	NN	O	O
early	NN	O	O
hematopoietic	NN	O	O
stages	NN	O	O
and	NN	O	O
during	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
to	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
or	NN	O	O
to	NN	O	O
neutrophils	NN	O	B-cell_type
has	NN	O	O
not	NN	O	O
been	NN	O	O
extensively	NN	O	O
investigated	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
PU.1	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
upregulated	NN	O	O
during	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
purified	NN	O	I-cell_line
CD34+	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
murine	NN	O	B-cell_line
multipotential	NN	O	I-cell_line
FDCP-mix	NN	O	I-cell_line
A4	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
PU.1	NN	O	B-protein
is	NN	O	O
upregulated	NN	O	O
as	NN	O	O
an	NN	O	O
early	NN	O	O
event	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
multipotential	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

PU.1	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
maintained	NN	O	O
at	NN	O	O
stable	NN	O	O
levels	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
U937	NN	O	B-cell_line
and	NN	O	O
HL-60	NN	O	B-cell_line
to	NN	O	O
monocytic	NN	O	B-cell_type
and	NN	O	I-cell_type
neutrophilic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

PU.1	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
highest	NN	O	O
levels	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
significant	NN	O	O
levels	NN	O	O
of	NN	O	O
Spi-B	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
are	NN	O	O
found	NN	O	O
only	NN	O	O
in	NN	O	O
some	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
spleen	NN	O	O
but	NN	O	O
are	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
or	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
translated	NN	O	O
Spi-B	NN	O	B-protein
protein	NN	O	I-protein
can	NN	O	O
bind	NN	O	O
to	NN	O	O
PU.1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
myeloid	NN	O	B-DNA
promoters	NN	O	I-DNA
and	NN	O	O
transactivate	NN	O	O
these	NN	O	O
promoters	NN	O	B-DNA
in	NN	O	O
nonmyeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
although	NN	O	O
PU.1	NN	O	B-protein
and	NN	O	O
Spi-B	NN	O	B-protein
may	NN	O	O
bind	NN	O	O
to	NN	O	O
similar	NN	O	O
DNA	NN	O	O
control	NN	O	O
elements	NN	O	O
and	NN	O	O
have	NN	O	O
redundancy	NN	O	O
of	NN	O	O
transactivation	NN	O	O
function	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
significant	NN	O	O
levels	NN	O	O
of	NN	O	O
Spi-B	NN	O	B-protein
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
makes	NN	O	O
it	NN	O	O
unlikely	NN	O	O
that	NN	O	O
Spi-B	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
significant	NN	O	O
role	NN	O	O
in	NN	O	O
myeloid	NN	O	O
lineage	NN	O	O
development	NN	O	O
and	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
PU.1	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
not	NN	O	O
only	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
but	NN	O	O
also	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
PU.1	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
both	NN	O	O
major	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
major	NN	O	O
positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
located	NN	O	O
5	NN	O	O
'	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
zeta-globin	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
zeta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
studied	NN	O	O
using	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
zeta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
deletion	NN	O	I-DNA
constructs	NN	O	I-DNA
to	NN	O	O
drive	NN	O	O
luciferase	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
transiently	NN	O	B-cell_line
transfected	NN	O	I-cell_line
human	NN	O	I-cell_line
erythroleukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
promoters	NN	O	B-DNA
were	NN	O	O
used	NN	O	O
without	NN	O	O
enhancers	NN	O	B-DNA
,	NN	O	O
or	NN	O	O
with	NN	O	O
enhancers	NN	O	B-DNA
derived	NN	O	O
from	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
locus	NN	O	I-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
alpha-globin	NN	O	B-DNA
HS-40	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
transfected	NN	O	O
into	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
express	NN	O	O
zeta-globin	NN	O	B-protein
,	NN	O	O
comparable	NN	O	O
amounts	NN	O	O
of	NN	O	O
activity	NN	O	O
were	NN	O	O
obtained	NN	O	O
from	NN	O	O
the	NN	O	O
-557	NN	O	B-DNA
and	NN	O	I-DNA
-417	NN	O	I-DNA
zeta-luciferase	NN	O	I-DNA
constructs	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
alpha-luciferase	NN	O	B-DNA
constructs	NN	O	I-DNA
when	NN	O	O
no	NN	O	O
enhancers	NN	O	B-DNA
or	NN	O	O
the	NN	O	O
alpha-globin	NN	O	B-DNA
locus	NN	O	I-DNA
enhancers	NN	O	I-DNA
were	NN	O	O
used	NN	O	O
.	NN	O	O

When	NN	O	O
the	NN	O	O
constructs	NN	O	O
were	NN	O	O
transfected	NN	O	O
into	NN	O	O
OCIM1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
zeta-globin	NN	O	B-protein
,	NN	O	O
the	NN	O	O
zeta-globin	NN	O	B-DNA
promoters	NN	O	I-DNA
were	NN	O	O
at	NN	O	O
best	NN	O	O
20	NN	O	O
%	NN	O	O
as	NN	O	O
active	NN	O	O
as	NN	O	O
the	NN	O	O
alpha-globin	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
sequences	NN	O	B-DNA
from	NN	O	I-DNA
-417	NN	O	I-DNA
to	NN	O	I-DNA
-207	NN	O	I-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
zeta-globin	NN	O	B-RNA
mRNA	NN	O	I-RNA
cap	NN	O	I-RNA
site	NN	O	I-RNA
were	NN	O	O
deleted	NN	O	O
,	NN	O	O
up	NN	O	O
to	NN	O	O
95	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
zeta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
was	NN	O	O
lost	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Reinsertion	NN	O	O
of	NN	O	O
these	NN	O	O
sequences	NN	O	O
into	NN	O	O
zeta-luciferase	NN	O	B-DNA
constructs	NN	O	I-DNA
missing	NN	O	O
the	NN	O	O
-417	NN	O	O
to	NN	O	O
-207	NN	O	O
region	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
sequences	NN	O	O
lack	NN	O	O
classical	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
.	NN	O	O

Point	NN	O	O
mutation	NN	O	O
of	NN	O	O
a	NN	O	O
GATA-1	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
-230	NN	O	O
reduced	NN	O	O
promoter	NN	O	O
activity	NN	O	O
by	NN	O	O
37	NN	O	O
%	NN	O	O
.	NN	O	O

Point	NN	O	O
mutation	NN	O	O
of	NN	O	O
a	NN	O	O
CCACC	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
-240	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
-230	NN	O	B-DNA
GATA-1	NN	O	I-DNA
site	NN	O	I-DNA
has	NN	O	O
a	NN	O	O
relatively	NN	O	O
low	NN	O	O
affinity	NN	O	O
for	NN	O	O
GATA-1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
experiments	NN	O	O
show	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
strong	NN	O	O
positive-acting	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
located	NN	O	O
between	NN	O	O
-417	NN	O	B-DNA
and	NN	O	I-DNA
-207	NN	O	I-DNA
bp	NN	O	I-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
zeta-globin	NN	O	B-DNA
mRNA	NN	O	I-DNA
cap	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
high-level	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
element	NN	O	O
requires	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
additional	NN	O	B-protein
unknown	NN	O	I-protein
factors	NN	O	I-protein
for	NN	O	O
maximal	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C-alpha	NN	O	I-protein
,	NN	O	I-protein
-epsilon	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
-zeta	NN	O	I-protein
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
multiple	NN	O	O
isotypes	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
and	NN	O	O
although	NN	O	O
it	NN	O	O
is	NN	O	O
well	NN	O	O
accepted	NN	O	O
that	NN	O	O
PKCs	NN	O	B-protein
have	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
individual	NN	O	O
PKC	NN	O	B-protein
isotypes	NN	O	I-protein
.	NN	O	O

To	NN	O	O
address	NN	O	O
this	NN	O	O
issue	NN	O	O
,	NN	O	O
mutationally	NN	O	O
active	NN	O	O
PKC-alpha	NN	O	B-protein
,	NN	O	I-protein
-epsilon	NN	O	I-protein
,	NN	O	I-protein
or	NN	O	I-protein
-zeta	NN	O	I-protein
have	NN	O	O
been	NN	O	O
transfected	NN	O	O
into	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
consequences	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
determined	NN	O	O
.	NN	O	O

p21ras	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Accordingly	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
constitutively	NN	O	B-protein
active	NN	O	I-protein
PKCs	NN	O	I-protein
were	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
mutationally	NN	O	O
activated	NN	O	O
p21ras	NN	O	B-protein
.	NN	O	O

The	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
PKC-epsilon	NN	O	B-protein
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
PKC-alpha	NN	O	B-protein
but	NN	O	O
not	NN	O	O
-zeta	NN	O	B-protein
,	NN	O	O
can	NN	O	O
regulate	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP-1	NN	O	B-protein
and	NN	O	O
nuclear	NN	O	O
factor	NN	O	B-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
PKC-epsilon	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
transactivation	NN	O	O
of	NN	O	O
NF-AT-1	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
was	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
stimulatory	NN	O	O
effect	NN	O	O
of	NN	O	O
a	NN	O	O
constitutively	NN	O	B-protein
activated	NN	O	I-protein
p21ras	NN	O	I-protein
.	NN	O	O

PKC-epsilon	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
PKC-alpha	NN	O	B-protein
nor	NN	O	O
activated	NN	O	O
p21ras	NN	O	B-protein
,	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
NF-KB	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Phorbol	NN	O	O
esters	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
CD69	NN	O	B-protein
whereas	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
activated	NN	O	B-protein
PKC	NN	O	I-protein
isotypes	NN	O	I-protein
tested	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
have	NN	O	O
this	NN	O	O
effect	NN	O	O
.	NN	O	O

Activated	NN	O	O
Src	NN	O	B-protein
and	NN	O	O
p21ras	NN	O	B-protein
were	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
CD69	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
selective	NN	O	O
functions	NN	O	O
for	NN	O	O
different	NN	O	O
PKC	NN	O	B-protein
isotypes	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
data	NN	O	O
comparing	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
activated	NN	O	B-protein
Ras	NN	O	I-protein
and	NN	O	O
PKC	NN	O	B-protein
mutants	NN	O	I-protein
suggest	NN	O	O
that	NN	O	O
PKC-alpha	NN	O	B-protein
,	NN	O	O
p21ras	NN	O	B-protein
,	NN	O	O
and	NN	O	O
PKC-epsilon	NN	O	B-protein
are	NN	O	O
not	NN	O	O
positioned	NN	O	O
linearly	NN	O	O
on	NN	O	O
a	NN	O	O
single	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differences	NN	O	O
in	NN	O	O
binding	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
to	NN	O	O
DNA	NN	O	O
in	NN	O	O
steroid-resistant	NN	O	O
asthma	NN	O	O
.	NN	O	O

Although	NN	O	O
glucocorticosteroids	NN	O	O
are	NN	O	O
a	NN	O	O
very	NN	O	O
effective	NN	O	O
treatment	NN	O	O
for	NN	O	O
asthma	NN	O	O
and	NN	O	O
other	NN	O	O
chronic	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
,	NN	O	O
a	NN	O	O
small	NN	O	O
proportion	NN	O	O
of	NN	O	O
patients	NN	O	O
are	NN	O	O
resistant	NN	O	O
to	NN	O	O
their	NN	O	O
therapeutic	NN	O	O
effects	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
for	NN	O	O
this	NN	O	O
steroid	NN	O	O
resistance	NN	O	O
is	NN	O	O
unclear	NN	O	O
.	NN	O	O

Steroid	NN	O	O
resistance	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
pharmacokinetic	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
by	NN	O	O
a	NN	O	O
defect	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
steroids	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
nor	NN	O	O
by	NN	O	O
defective	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
this	NN	O	O
receptor	NN	O	O
,	NN	O	O
thereby	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
molecular	NN	O	O
abnormality	NN	O	O
lies	NN	O	O
distal	NN	O	O
to	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
nuclear	NN	O	O
translocated	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
to	NN	O	O
bind	NN	O	O
to	NN	O	O
their	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
(	NN	O	O
GRE	NN	O	B-DNA
)	NN	O	O
using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
steroid-sensitive	NN	O	O
and	NN	O	O
steroid-resistant	NN	O	O
asthma	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
to	NN	O	O
DNA	NN	O	O
in	NN	O	O
these	NN	O	O
patients	NN	O	O
was	NN	O	O
also	NN	O	O
studied	NN	O	O
using	NN	O	O
Scatchard	NN	O	O
analysis	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
induced	NN	O	O
a	NN	O	O
significant	NN	O	O
rapid	NN	O	O
and	NN	O	O
sustained	NN	O	O
twofold	NN	O	O
increase	NN	O	O
in	NN	O	O
GRE	NN	O	B-DNA
binding	NN	O	O
in	NN	O	O
PBMCs	NN	O	B-cell_type
from	NN	O	O
steroid-sensitive	NN	O	O
asthmatic	NN	O	O
patients	NN	O	O
and	NN	O	O
nonasthmatic	NN	O	O
individuals	NN	O	O
,	NN	O	O
but	NN	O	O
this	NN	O	O
was	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
in	NN	O	O
steroid-resistant	NN	O	O
asthmatic	NN	O	O
patients	NN	O	O
.	NN	O	O

Scatchard	NN	O	O
analysis	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
-	NN	O	O
GRE	NN	O	B-DNA
binding	NN	O	O
showed	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
binding	NN	O	O
affinity	NN	O	O
but	NN	O	O
did	NN	O	O
show	NN	O	O
a	NN	O	O
reduced	NN	O	O
number	NN	O	O
of	NN	O	O
receptors	NN	O	B-protein
available	NN	O	O
for	NN	O	O
DNA	NN	O	O
binding	NN	O	O
in	NN	O	O
the	NN	O	O
steroid-resistant	NN	O	O
patients	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
to	NN	O	O
bind	NN	O	O
to	NN	O	O
GRE	NN	O	B-DNA
is	NN	O	O
impaired	NN	O	O
in	NN	O	O
steroid-resistant	NN	O	O
patients	NN	O	O
because	NN	O	O
of	NN	O	O
a	NN	O	O
reduced	NN	O	O
number	NN	O	O
of	NN	O	O
receptors	NN	O	B-protein
available	NN	O	O
for	NN	O	O
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin-5	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
involves	NN	O	O
JAK2	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
Stat1	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

Signaling	NN	O	O
by	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
including	NN	O	O
interferons	NN	O	B-protein
,	NN	O	O
interleukins	NN	O	B-protein
,	NN	O	O
and	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
involves	NN	O	O
activation	NN	O	O
of	NN	O	O
JAK	NN	O	B-protein
kinases	NN	O	I-protein
and	NN	O	O
Stat	NN	O	B-protein
(	NN	O	I-protein
Signal	NN	O	I-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

At	NN	O	O
present	NN	O	O
,	NN	O	O
not	NN	O	O
much	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
interleukin-5	NN	O	B-protein
(	NN	O	O
IL-5	NN	O	B-protein
)	NN	O	O
exerts	NN	O	O
its	NN	O	O
diverse	NN	O	O
biologic	NN	O	O
effects	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
are	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
important	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
IL-5	NN	O	B-protein
and	NN	O	O
were	NN	O	O
used	NN	O	O
here	NN	O	O
to	NN	O	O
study	NN	O	O
IL-5	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
a	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

IL-5	NN	O	B-protein
induced	NN	O	O
rapid	NN	O	O
and	NN	O	O
transient	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
JAK2	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
IL-5	NN	O	B-protein
induced	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
DNA-binding	NN	O	B-protein
complexes	NN	O	I-protein
,	NN	O	O
using	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
normal	NN	O	O
human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
IL-6/interferon-gamma	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
ICAM-1	NN	O	B-DNA
pIRE	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
an	NN	O	O
electromobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

From	NN	O	O
supershift	NN	O	O
experiments	NN	O	O
it	NN	O	O
was	NN	O	O
concluded	NN	O	O
that	NN	O	O
one	NN	O	O
DNA-binding	NN	O	B-protein
complex	NN	O	I-protein
contained	NN	O	O
Stat1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
probably	NN	O	O
as	NN	O	O
a	NN	O	O
homodimer	NN	O	O
.	NN	O	O

Both	NN	O	O
DNA-binding	NN	O	B-protein
complexes	NN	O	I-protein
were	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
phosphotyrosine	NN	O	B-protein
antibody	NN	O	I-protein
(	NN	O	O
4G10	NN	O	B-protein
)	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
complex	NN	O	O
formation	NN	O	O
.	NN	O	O

IL-3	NN	O	B-protein
and	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
induced	NN	O	O
,	NN	O	O
similar	NN	O	O
to	NN	O	O
IL-5	NN	O	B-protein
,	NN	O	O
two	NN	O	O
DNA-binding	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
,	NN	O	O
including	NN	O	O
Stat1	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
that	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
involve	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
JAK	NN	O	B-protein
kinase	NN	O	I-protein
family	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Stat	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Infection	NN	O	O
and	NN	O	O
replication	NN	O	O
of	NN	O	O
Tat-	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
viruses	NN	O	O
:	NN	O	O
genetic	NN	O	O
analyses	NN	O	O
of	NN	O	O
LTR	NN	O	B-DNA
and	NN	O	O
tat	NN	O	B-protein
mutations	NN	O	I-protein
in	NN	O	O
primary	NN	O	B-cell_type
and	NN	O	I-cell_type
long-term	NN	O	I-cell_type
human	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Tat	NN	O	B-protein
is	NN	O	O
an	NN	O	O
essential	NN	O	O
regulatory	NN	O	B-protein
protein	NN	O	I-protein
for	NN	O	O
the	NN	O	O
replication	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
tat	NN	O	B-DNA
gene	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
block	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Several	NN	O	O
studies	NN	O	O
have	NN	O	O
established	NN	O	O
that	NN	O	O
Tat	NN	O	B-protein
releases	NN	O	O
an	NN	O	O
elongation	NN	O	B-protein
block	NN	O	I-protein
to	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
whether	NN	O	O
this	NN	O	O
mechanism	NN	O	O
alone	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
explain	NN	O	O
the	NN	O	O
block	NN	O	O
to	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
when	NN	O	O
Tat	NN	O	B-protein
is	NN	O	O
absent	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
possible	NN	O	O
that	NN	O	O
Tat	NN	O	B-protein
is	NN	O	O
also	NN	O	O
needed	NN	O	O
for	NN	O	O
other	NN	O	O
functions	NN	O	O
during	NN	O	O
HIV	NN	O	O
replication	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
these	NN	O	O
hypotheses	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
several	NN	O	O
tat	NN	O	B-protein
mutants	NN	O	I-protein
,	NN	O	O
including	NN	O	O
two	NN	O	O
stop	NN	O	B-protein
codon	NN	O	I-protein
mutants	NN	O	I-protein
and	NN	O	O
one	NN	O	O
deletion	NN	O	B-protein
mutant	NN	O	I-protein
using	NN	O	O
replication-competent	NN	O	B-DNA
HIV-1	NN	O	I-DNA
constructs	NN	O	I-DNA
carrying	NN	O	O
wild-type	NN	O	B-DNA
or	NN	O	I-DNA
mutant	NN	O	I-DNA
LTRs	NN	O	I-DNA
with	NN	O	O
modifications	NN	O	O
in	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
and/or	NN	O	I-DNA
Sp1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
Tat-	NN	O	O
HIV-1	NN	O	O
with	NN	O	O
wild-type	NN	O	B-DNA
LTRs	NN	O	I-DNA
can	NN	O	O
replicate	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
the	NN	O	O
virus	NN	O	O
produced	NN	O	O
from	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
can	NN	O	O
infect	NN	O	O
primary	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
propagation	NN	O	O
of	NN	O	O
the	NN	O	O
Tat	NN	O	B-protein
mutants	NN	O	I-protein
containing	NN	O	O
wild-type	NN	O	B-DNA
LTRs	NN	O	I-DNA
was	NN	O	O
less	NN	O	O
efficient	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
LTR-modified	NN	O	B-protein
Tat	NN	O	I-protein
mutants	NN	O	I-protein
.	NN	O	O

Large	NN	O	O
amounts	NN	O	O
of	NN	O	O
viral	NN	O	B-RNA
RNA	NN	O	I-RNA
and	NN	O	O
particles	NN	O	O
were	NN	O	O
synthesized	NN	O	O
in	NN	O	O
infections	NN	O	O
established	NN	O	O
using	NN	O	O
the	NN	O	O
tat	NN	O	B-protein
mutants	NN	O	I-protein
that	NN	O	O
contain	NN	O	O
modified	NN	O	B-DNA
LTRs	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
efficient	NN	O	O
propagation	NN	O	O
of	NN	O	O
the	NN	O	O
LTR-modified	NN	O	B-protein
tat	NN	O	I-protein
mutants	NN	O	I-protein
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
some	NN	O	O
lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
have	NN	O	O
been	NN	O	O
transformed	NN	O	O
with	NN	O	O
other	NN	O	O
viruses	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
despite	NN	O	O
its	NN	O	O
essential	NN	O	O
role	NN	O	O
for	NN	O	O
releasing	NN	O	O
an	NN	O	O
elongation	NN	O	B-protein
block	NN	O	I-protein
,	NN	O	O
Tat	NN	O	B-protein
is	NN	O	O
not	NN	O	O
otherwise	NN	O	O
absolutely	NN	O	O
required	NN	O	O
for	NN	O	O
synthesis	NN	O	O
of	NN	O	O
full-length	NN	O	B-RNA
HIV	NN	O	I-RNA
transcripts	NN	O	I-RNA
and	NN	O	O
assembly	NN	O	O
of	NN	O	O
virus	NN	O	O
particles	NN	O	O
.	NN	O	O

Direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
genomes	NN	O	I-DNA
and	NN	O	O
reinfection	NN	O	O
kinetics	NN	O	O
showed	NN	O	O
no	NN	O	O
evidence	NN	O	O
of	NN	O	O
wild-type	NN	O	O
reversion	NN	O	O
even	NN	O	O
after	NN	O	O
prolonged	NN	O	O
infection	NN	O	O
with	NN	O	O
the	NN	O	O
Tat	NN	O	B-protein
-virus	NN	O	O
.	NN	O	O

The	NN	O	O
implications	NN	O	O
for	NN	O	O
in	NN	O	O
vivo	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
and	NN	O	O
potential	NN	O	O
application	NN	O	O
of	NN	O	O
this	NN	O	O
system	NN	O	O
to	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
alternative	NN	O	O
Tat	NN	O	B-protein
function	NN	O	O
are	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Oct-2A	NN	O	I-protein
and	NN	O	O
the	NN	O	O
high	NN	O	B-protein
mobility	NN	O	I-protein
group	NN	O	I-protein
protein	NN	O	I-protein
I/Y	NN	O	B-protein
in	NN	O	O
HLA-DRA	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

The	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
major	NN	O	I-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
gene	NN	O	I-DNA
HLA-DRA	NN	O	B-DNA
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
in	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
HLA-DRA	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
is	NN	O	O
inducible	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
by	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
lymphoid-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Oct-2A	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
HLA-DRA	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
class	NN	O	B-cell_line
II-positive	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
high	NN	O	B-protein
mobility	NN	O	I-protein
group	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
HMG	NN	O	B-protein
)	NN	O	O
I/Y	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
multiple	NN	O	O
sites	NN	O	O
within	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
the	NN	O	O
Oct-2A	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Coexpression	NN	O	O
of	NN	O	O
HMG	NN	O	B-protein
I/Y	NN	O	I-protein
and	NN	O	O
Oct-2	NN	O	B-protein
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
lacking	NN	O	O
Oct-2	NN	O	B-protein
results	NN	O	O
in	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
HLA-DRA	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
DNA-binding	NN	O	O
studies	NN	O	O
reveal	NN	O	O
that	NN	O	O
HMG	NN	O	O
I/Y	NN	O	B-protein
stimulates	NN	O	O
Oct-2A	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
HLA-DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Oct-2A	NN	O	B-protein
and	NN	O	O
HMG	NN	O	B-protein
I/Y	NN	O	I-protein
may	NN	O	O
synergize	NN	O	O
to	NN	O	O
activate	NN	O	O
HLA-DRA	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
Oct-2A	NN	O	B-protein
is	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
the	NN	O	O
HLA-DRA	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
antisense	NN	O	B-DNA
HMG	NN	O	I-DNA
I/Y	NN	O	I-DNA
dramatically	NN	O	O
decreases	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
induction	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
distinct	NN	O	O
sets	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
modes	NN	O	O
of	NN	O	O
HLA-DRA	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
HMG	NN	O	B-protein
I/Y	NN	O	I-protein
may	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
IFN-gamma	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

-DOCSTART-	O

Heterogeneous	NN	O	O
expression	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
latent	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
endemic	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
.	NN	O	O

Epstein-Barr	NN	O	B-cell_type
virus	NN	O	I-cell_type
(	NN	O	I-cell_type
EBV	NN	O	I-cell_type
)	NN	O	I-cell_type
-infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
sustain	NN	O	O
three	NN	O	O
distinct	NN	O	O
forms	NN	O	O
of	NN	O	O
virus	NN	O	O
latency	NN	O	O
.	NN	O	O

In	NN	O	O
lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
six	NN	O	O
EBV-encoded	NN	O	B-protein
nuclear	NN	O	I-protein
antigens	NN	O	I-protein
(	NN	O	O
EBNA1	NN	O	B-protein
,	NN	O	I-protein
2	NN	O	I-protein
,	NN	O	I-protein
3A	NN	O	I-protein
,	NN	O	I-protein
3B	NN	O	I-protein
,	NN	O	I-protein
3C	NN	O	I-protein
,	NN	O	I-protein
-LP	NN	O	I-protein
)	NN	O	O
,	NN	O	O
three	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
LMP1	NN	O	B-protein
,	NN	O	O
2A	NN	O	B-protein
,	NN	O	O
2B	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
two	NN	O	O
nuclear	NN	O	B-RNA
RNAs	NN	O	I-RNA
(	NN	O	O
EBERs	NN	O	B-RNA
)	NN	O	O
are	NN	O	O
expressed	NN	O	O
.	NN	O	O

This	NN	O	O
form	NN	O	O
of	NN	O	O
latency	NN	O	O
,	NN	O	O
termed	NN	O	O
latency	NN	O	O
III	NN	O	O
,	NN	O	O
is	NN	O	O
also	NN	O	O
encountered	NN	O	O
in	NN	O	O
some	NN	O	O
posttransplant	NN	O	O
lymphoproliferative	NN	O	O
disorders	NN	O	O
.	NN	O	O

In	NN	O	O
EBV-positive	NN	O	O
cases	NN	O	O
of	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
,	NN	O	O
the	NN	O	O
EBERs	NN	O	B-RNA
,	NN	O	O
EBNA1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
LMPs	NN	O	B-protein
are	NN	O	O
expressed	NN	O	O
(	NN	O	O
latency	NN	O	O
II	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
BL	NN	O	O
)	NN	O	O
only	NN	O	O
the	NN	O	O
EBERs	NN	O	B-RNA
and	NN	O	O
EBNA1	NN	O	B-protein
have	NN	O	O
been	NN	O	O
detected	NN	O	O
(	NN	O	O
latency	NN	O	O
I	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
EBV	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
17	NN	O	O
cases	NN	O	O
of	NN	O	O
EBV-positive	NN	O	O
endemic	NN	O	O
BL	NN	O	O
by	NN	O	O
immunohistology	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
was	NN	O	O
seen	NN	O	O
in	NN	O	O
variable	NN	O	O
proportions	NN	O	O
of	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
two	NN	O	O
cases	NN	O	O
and	NN	O	O
EBNA2	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
some	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
three	NN	O	O
other	NN	O	O
cases	NN	O	O
.	NN	O	O

Also	NN	O	O
,	NN	O	O
the	NN	O	O
BZLF1	NN	O	B-protein
trans-activator	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
few	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
6	NN	O	O
cases	NN	O	O
,	NN	O	O
indicating	NN	O	O
entry	NN	O	O
into	NN	O	O
the	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
.	NN	O	O

A	NN	O	O
phenotypic	NN	O	O
drift	NN	O	O
from	NN	O	O
latency	NN	O	O
I	NN	O	O
to	NN	O	O
latency	NN	O	O
III	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
previously	NN	O	O
in	NN	O	O
some	NN	O	O
BL	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
similar	NN	O	O
phenomenon	NN	O	O
may	NN	O	O
occur	NN	O	O
in	NN	O	O
BL	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
operational	NN	O	O
definition	NN	O	O
of	NN	O	O
EBV	NN	O	O
latencies	NN	O	O
is	NN	O	O
not	NN	O	O
easily	NN	O	O
applied	NN	O	O
to	NN	O	O
human	NN	O	O
tumors	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
Jak	NN	O	B-protein
-STAT	NN	O	B-protein
1	NN	O	I-protein
signaling	NN	O	O
pathway	NN	O	O
by	NN	O	O
IL-5	NN	O	B-protein
in	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
intracellular	NN	O	O
signal	NN	O	O
transduction	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
in	NN	O	O
eosinophils	NN	O	B-cell_type
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

The	NN	O	O
objective	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
newly	NN	O	O
discovered	NN	O	O
Jak	NN	O	B-protein
-STAT	NN	O	B-protein
pathway	NN	O	O
in	NN	O	O
the	NN	O	O
IL-5	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Eosinophils	NN	O	B-cell_type
were	NN	O	O
purified	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
by	NN	O	O
discontinuous	NN	O	O
Percoll	NN	O	O
gradients	NN	O	O
and	NN	O	O
stimulated	NN	O	O
with	NN	O	O
IL-5	NN	O	B-protein
.	NN	O	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
Jak	NN	O	B-protein
2	NN	O	I-protein
was	NN	O	O
investigated	NN	O	O
by	NN	O	O
immunoprecipitation	NN	O	O
followed	NN	O	O
by	NN	O	O
immunoblotting	NN	O	O
for	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
Jak	NN	O	B-protein
2	NN	O	I-protein
was	NN	O	O
studied	NN	O	O
by	NN	O	O
autophosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
immunoprecipitated	NN	O	B-protein
kinase	NN	O	I-protein
.	NN	O	O

Jak	NN	O	B-protein
2	NN	O	I-protein
was	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
within	NN	O	O
1	NN	O	O
to	NN	O	O
3	NN	O	O
min	NN	O	O
after	NN	O	O
stimulation	NN	O	O
of	NN	O	O
eosinophils	NN	O	B-cell_type
with	NN	O	O
IL-5	NN	O	B-protein
.	NN	O	O

Further	NN	O	O
,	NN	O	O
the	NN	O	O
immunoprecipitated	NN	O	B-protein
Jak	NN	O	I-protein
2	NN	O	I-protein
obtained	NN	O	O
from	NN	O	O
IL-5-stimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
underwent	NN	O	O
autophosphorylation	NN	O	O
.	NN	O	O

Jak	NN	O	B-protein
2	NN	O	I-protein
coprecipitated	NN	O	O
with	NN	O	O
the	NN	O	O
beta-subunit	NN	O	O
of	NN	O	O
the	NN	O	O
IL-5	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
physical	NN	O	O
association	NN	O	O
of	NN	O	O
the	NN	O	O
kinase	NN	O	B-protein
with	NN	O	O
the	NN	O	O
receptor	NN	O	O
.	NN	O	O

The	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
STAT-1	NN	O	I-protein
(	NN	O	O
p91	NN	O	B-protein
)	NN	O	O
was	NN	O	O
investigated	NN	O	O
by	NN	O	O
immunoprecipitation	NN	O	O
followed	NN	O	O
by	NN	O	O
immunoblotting	NN	O	O
for	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

STAT-1	NN	O	B-protein
was	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
within	NN	O	O
15	NN	O	O
min	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
STAT-1	NN	O	B-protein
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
was	NN	O	O
studied	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

IL-5	NN	O	B-protein
induced	NN	O	O
two	NN	O	O
proteins	NN	O	O
that	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
gamma-activating	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
anti-STAT-1	NN	O	B-protein
Ab	NN	O	I-protein
,	NN	O	O
the	NN	O	O
band	NN	O	O
was	NN	O	O
supershifted	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
IL-5	NN	O	B-protein
activated	NN	O	O
the	NN	O	O
Jak	NN	O	B-protein
2	NN	O	I-protein
-STAT	NN	O	B-protein
1	NN	O	I-protein
signaling	NN	O	O
pathway	NN	O	O
in	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
speculate	NN	O	O
that	NN	O	O
the	NN	O	O
Jak	NN	O	B-protein
2	NN	O	I-protein
-STAT	NN	O	B-protein
1	NN	O	I-protein
pathway	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
IL-5-inducible	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Reduced	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
a	NN	O	O
prognostic	NN	O	O
parameter	NN	O	O
for	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
:	NN	O	O
a	NN	O	O
prospective	NN	O	O
longitudinal	NN	O	O
study	NN	O	O
.	NN	O	O

Immunosuppression	NN	O	O
has	NN	O	O
been	NN	O	O
often	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
malignant	NN	O	O
diseases	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phytohaemagglutinin	NN	O	B-protein
(	NN	O	O
PHA	NN	O	B-protein
)	NN	O	O
was	NN	O	O
assessed	NN	O	O
prospectively	NN	O	O
in	NN	O	O
90	NN	O	O
patients	NN	O	O
with	NN	O	O
stage	NN	O	O
I-III	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

Whereas	NN	O	O
PHA	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
of	NN	O	O
PBMCs	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
preoperatively	NN	O	O
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
data	NN	O	O
obtained	NN	O	O
in	NN	O	O
healthy	NN	O	O
control	NN	O	O
individuals	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
the	NN	O	O
defect	NN	O	O
in	NN	O	O
PHA	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
of	NN	O	O
PBMCs	NN	O	B-cell_type
depended	NN	O	O
upon	NN	O	O
the	NN	O	O
tumour	NN	O	O
burden	NN	O	O
as	NN	O	O
manifested	NN	O	O
by	NN	O	O
tumour	NN	O	O
size	NN	O	O
and	NN	O	O
axillary	NN	O	O
lymph	NN	O	O
node	NN	O	O
involvement	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.003	NN	O	O
in	NN	O	O
each	NN	O	O
case	NN	O	O
)	NN	O	O
.	NN	O	O

PHA	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
of	NN	O	O
PBMCs	NN	O	B-cell_type
dropped	NN	O	O
significantly	NN	O	O
in	NN	O	O
patients	NN	O	O
who	NN	O	O
received	NN	O	O
adjuvant	NN	O	O
chemotherapy	NN	O	O
consisting	NN	O	O
of	NN	O	O
cyclophosphamide	NN	O	O
,	NN	O	O
methotrexate	NN	O	O
and	NN	O	O
fluorouracil	NN	O	O
(	NN	O	O
CMF	NN	O	O
)	NN	O	O
after	NN	O	O
an	NN	O	O
observation	NN	O	O
period	NN	O	O
of	NN	O	O
6	NN	O	O
months	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
patients	NN	O	O
under	NN	O	O
adjuvant	NN	O	O
treatment	NN	O	O
with	NN	O	O
tamoxifen	NN	O	O
only	NN	O	O
.	NN	O	O

After	NN	O	O
an	NN	O	O
additional	NN	O	O
6	NN	O	O
months	NN	O	O
(	NN	O	O
i.e.	NN	O	O
12	NN	O	O
months	NN	O	O
after	NN	O	O
surgery	NN	O	O
)	NN	O	O
,	NN	O	O
PHA	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
of	NN	O	O
PBMCs	NN	O	B-cell_type
was	NN	O	O
similar	NN	O	O
in	NN	O	O
patients	NN	O	O
after	NN	O	O
adjuvant	NN	O	O
chemotherapy	NN	O	O
with	NN	O	O
CMF	NN	O	O
and	NN	O	O
in	NN	O	O
those	NN	O	O
receiving	NN	O	O
continued	NN	O	O
adjuvant	NN	O	O
tamoxifen	NN	O	O
treatment	NN	O	O
(	NN	O	O
P	NN	O	O
>	NN	O	O
0.1	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
in	NN	O	O
all	NN	O	O
patients	NN	O	O
still	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
healthy	NN	O	O
controls	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
.	NN	O	O

When	NN	O	O
data	NN	O	O
obtained	NN	O	O
preoperatively	NN	O	O
and	NN	O	O
after	NN	O	O
12	NN	O	O
months	NN	O	O
were	NN	O	O
compared	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
out	NN	O	O
of	NN	O	O
23	NN	O	O
patients	NN	O	O
whose	NN	O	O
PBMCs	NN	O	B-cell_type
had	NN	O	O
experienced	NN	O	O
a	NN	O	O
drop	NN	O	O
in	NN	O	O
their	NN	O	O
PHA	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
,	NN	O	O
14	NN	O	O
(	NN	O	O
61	NN	O	O
%	NN	O	O
)	NN	O	O
had	NN	O	O
developed	NN	O	O
metastatic	NN	O	O
disease	NN	O	O
within	NN	O	O
the	NN	O	O
subsequent	NN	O	O
24	NN	O	O
months	NN	O	O
(	NN	O	O
i.e.	NN	O	O
36	NN	O	O
months	NN	O	O
after	NN	O	O
surgery	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
out	NN	O	O
of	NN	O	O
59	NN	O	O
patients	NN	O	O
whose	NN	O	O
PBMCs	NN	O	B-cell_type
showed	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
their	NN	O	O
PHA	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
within	NN	O	O
the	NN	O	O
first	NN	O	O
12	NN	O	O
months	NN	O	O
after	NN	O	O
surgery	NN	O	O
,	NN	O	O
only	NN	O	O
one	NN	O	O
(	NN	O	O
2	NN	O	O
%	NN	O	O
)	NN	O	O
presented	NN	O	O
with	NN	O	O
disease	NN	O	O
progression	NN	O	O
.	NN	O	O

We	NN	O	O
thus	NN	O	O
conclude	NN	O	O
that	NN	O	O
PHA	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
of	NN	O	O
PBMCs	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
depends	NN	O	O
upon	NN	O	O
the	NN	O	O
tumour	NN	O	O
load	NN	O	O
and	NN	O	O
is	NN	O	O
a	NN	O	O
good	NN	O	O
clinical	NN	O	O
predictor	NN	O	O
for	NN	O	O
the	NN	O	O
further	NN	O	O
course	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
transcription	NN	O	O
by	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-E1	NN	O	B-protein
(	NN	O	O
YY1	NN	O	B-protein
)	NN	O	O
to	NN	O	O
a	NN	O	O
newly	NN	O	O
identified	NN	O	O
element	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	O
exon	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
DR	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
previously	NN	O	O
unrecognized	NN	O	O
element	NN	O	O
,	NN	O	O
located	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
start	NN	O	B-DNA
site	NN	O	I-DNA
of	NN	O	I-DNA
transcription	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
first	NN	O	O
exon	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
DR	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
has	NN	O	O
been	NN	O	O
defined	NN	O	O
that	NN	O	O
enhances	NN	O	O
promoter	NN	O	O
activity	NN	O	O
up	NN	O	O
to	NN	O	O
eightfold	NN	O	O
in	NN	O	O
a	NN	O	O
position-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
this	NN	O	O
DNA-binding	NN	O	B-DNA
site	NN	O	I-DNA
abolished	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
human	NN	O	O
B	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
and	NN	O	O
decreased	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
DR	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
to	NN	O	O
a	NN	O	O
basal	NN	O	O
level	NN	O	O
.	NN	O	O

Significant	NN	O	O
sequence	NN	O	O
homology	NN	O	O
of	NN	O	O
this	NN	O	O
element	NN	O	B-DNA
was	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
of	NN	O	O
the	NN	O	O
DR	NN	O	B-DNA
beta	NN	O	I-DNA
,	NN	O	I-DNA
DP	NN	O	I-DNA
alpha	NN	O	I-DNA
and	NN	O	I-DNA
-beta	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
DQ	NN	O	I-DNA
alpha	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
always	NN	O	O
located	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
DR	NN	O	B-DNA
alpha	NN	O	I-DNA
and	NN	O	I-DNA
DP	NN	O	I-DNA
alpha	NN	O	I-DNA
element	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
to	NN	O	O
the	NN	O	O
element	NN	O	O
in	NN	O	O
the	NN	O	O
DQ	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
NF-E1	NN	O	B-protein
(	NN	O	O
YY1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

This	NN	O	O
protein	NN	O	O
,	NN	O	O
previously	NN	O	O
identified	NN	O	O
by	NN	O	O
its	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
kappa	NN	O	I-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
mu	NN	O	I-DNA
E1	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
thus	NN	O	O
also	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
quite	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
core	NN	O	B-protein
binding	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	I-protein
AML1	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
beta	NN	O	I-protein
subunits	NN	O	O
on	NN	O	O
murine	NN	O	B-DNA
leukemia	NN	O	I-DNA
virus	NN	O	I-DNA
enhancer	NN	O	I-DNA
cores	NN	O	I-DNA
.	NN	O	O

Core	NN	O	B-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
CBF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
polyomavirus	NN	O	B-protein
enhancer-binding	NN	O	I-protein
protein	NN	O	I-protein
2	NN	O	I-protein
and	NN	O	O
SL3	NN	O	B-protein
enhancer	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
is	NN	O	O
a	NN	O	O
mammalian	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
to	NN	O	O
an	NN	O	O
element	NN	O	O
termed	NN	O	O
the	NN	O	O
core	NN	O	O
within	NN	O	O
the	NN	O	O
enhancers	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
murine	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
family	NN	O	O
of	NN	O	O
retroviruses	NN	O	O
.	NN	O	O

The	NN	O	O
core	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
SL3	NN	O	O
virus	NN	O	O
are	NN	O	O
important	NN	O	O
genetic	NN	O	O
determinants	NN	O	O
of	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
this	NN	O	O
virus	NN	O	O
to	NN	O	O
induce	NN	O	O
T-cell	NN	O	O
lymphomas	NN	O	O
and	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

CBF	NN	O	B-protein
consists	NN	O	O
of	NN	O	O
two	NN	O	O
subunits	NN	O	O
,	NN	O	O
a	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
subunit	NN	O	I-protein
,	NN	O	O
CBF	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
second	NN	O	O
subunit	NN	O	O
,	NN	O	O
CBF	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
that	NN	O	O
stimulates	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
CBF	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
that	NN	O	O
encodes	NN	O	O
a	NN	O	O
CBF	NN	O	B-protein
alpha	NN	O	I-protein
subunit	NN	O	I-protein
is	NN	O	O
AML1	NN	O	B-protein
,	NN	O	O
also	NN	O	O
called	NN	O	O
Cbf	NN	O	B-protein
alpha	NN	O	I-protein
2	NN	O	I-protein
.	NN	O	O

This	NN	O	O
locus	NN	O	O
is	NN	O	O
rearranged	NN	O	O
by	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
in	NN	O	O
human	NN	O	O
myeloproliferative	NN	O	O
disorders	NN	O	O
and	NN	O	O
leukemias	NN	O	O
.	NN	O	O

An	NN	O	O
exogenously	NN	O	O
expressed	NN	O	O
Cbf	NN	O	B-protein
alpha	NN	O	I-protein
2-encoded	NN	O	I-protein
subunit	NN	O	I-protein
(	NN	O	O
CBF	NN	O	B-protein
alpha	NN	O	I-protein
2-451	NN	O	I-protein
)	NN	O	O
stimulated	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
SL3	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
P19	NN	O	B-protein
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Activity	NN	O	O
was	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

Three	NN	O	O
different	NN	O	O
isoforms	NN	O	O
of	NN	O	O
CBF	NN	O	B-protein
beta	NN	O	I-protein
were	NN	O	O
also	NN	O	O
tested	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
on	NN	O	O
the	NN	O	O
SL3	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
longest	NN	O	O
form	NN	O	O
,	NN	O	O
CBF	NN	O	B-protein
beta-187	NN	O	I-protein
,	NN	O	O
increased	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
stimulation	NN	O	O
by	NN	O	O
CBF	NN	O	B-protein
alpha	NN	O	I-protein
2-451	NN	O	I-protein
twofold	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
it	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
in	NN	O	O
P19	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
CBF	NN	O	B-protein
beta	NN	O	I-protein
required	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
CBF	NN	O	B-protein
alpha	NN	O	I-protein
subunit	NN	O	I-protein
,	NN	O	O
as	NN	O	O
a	NN	O	O
form	NN	O	O
of	NN	O	O
CBF	NN	O	B-protein
beta	NN	O	I-protein
that	NN	O	O
lacked	NN	O	O
binding	NN	O	O
ability	NN	O	O
,	NN	O	O
CBF	NN	O	B-protein
beta-148	NN	O	I-protein
,	NN	O	O
failed	NN	O	O
to	NN	O	O
increase	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicated	NN	O	O
that	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
certain	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
the	NN	O	O
maximum	NN	O	O
activity	NN	O	O
of	NN	O	O
CBF	NN	O	B-protein
required	NN	O	O
both	NN	O	O
subunits	NN	O	O
.	NN	O	O

They	NN	O	O
also	NN	O	O
provided	NN	O	O
support	NN	O	O
for	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
CBF	NN	O	B-protein
is	NN	O	O
a	NN	O	O
factor	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
that	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
recognition	NN	O	O
of	NN	O	O
the	NN	O	O
SL3	NN	O	B-DNA
cores	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
also	NN	O	O
examined	NN	O	O
whether	NN	O	O
CBF	NN	O	B-protein
could	NN	O	O
distinguish	NN	O	O
a	NN	O	O
1-bp	NN	O	O
difference	NN	O	O
between	NN	O	O
the	NN	O	O
enhancer	NN	O	O
core	NN	O	O
of	NN	O	O
SL3	NN	O	O
and	NN	O	O
the	NN	O	O
core	NN	O	O
of	NN	O	O
the	NN	O	O
nonleukemogenic	NN	O	O
virus	NN	O	O
,	NN	O	O
Akv	NN	O	O
.	NN	O	O

This	NN	O	O
difference	NN	O	O
strongly	NN	O	O
affects	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	O
cells	NN	O	O
and	NN	O	O
leukemogenicity	NN	O	O
of	NN	O	O
SL3	NN	O	B-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
no	NN	O	O
combination	NN	O	O
of	NN	O	O
CBF	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
CBF	NN	O	B-protein
beta	NN	O	I-protein
subunits	NN	O	O
that	NN	O	O
we	NN	O	O
tested	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
distinguish	NN	O	O
the	NN	O	O
1-bp	NN	O	O
difference	NN	O	O
in	NN	O	O
transcription	NN	O	O
assays	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
a	NN	O	O
complete	NN	O	O
understanding	NN	O	O
of	NN	O	O
how	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
recognize	NN	O	O
the	NN	O	O
SL3	NN	O	B-DNA
core	NN	O	I-DNA
remains	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-10	NN	O	I-protein
inhibits	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

IL-10	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
suppress	NN	O	O
cytokine	NN	O	B-protein
synthesis	NN	O	O
by	NN	O	O
different	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
studies	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-10	NN	O	I-protein
inhibits	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-stimulated	NN	O	O
production	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
-alpha	NN	O	I-protein
by	NN	O	O
blocking	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
,	NN	O	O
we	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
monocytes	NN	O	O
stimulated	NN	O	O
with	NN	O	O
LPS	NN	O	O
or	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
IL-10	NN	O	B-protein
inhibits	NN	O	O
nuclear	NN	O	O
stimulation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-DNA
cytokine	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O
Several	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
including	NN	O	O
NF-	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
AP-2	NN	O	B-protein
,	NN	O	O
GR	NN	O	B-protein
,	NN	O	O
CREB	NN	O	B-protein
,	NN	O	O
Oct-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Sp-1	NN	O	B-protein
are	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

This	NN	O	O
selective	NN	O	O
inhibition	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
occurs	NN	O	O
rapidly	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
and	NN	O	O
correlates	NN	O	O
well	NN	O	O
with	NN	O	O
IL-10	NN	O	B-protein
's	NN	O	O
cytokine	NN	O	B-protein
synthesis	NN	O	O
inhibitory	NN	O	O
activity	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
both	NN	O	O
kinetics	NN	O	O
and	NN	O	O
dose	NN	O	O
responsiveness	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
compounds	NN	O	O
such	NN	O	O
as	NN	O	O
tosylphenylalanyl	NN	O	O
chloromethyl	NN	O	O
ketone	NN	O	O
and	NN	O	O
pyrrolidinedithiocarbamate	NN	O	O
that	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
selectively	NN	O	O
inhibit	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
block	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
LPS-stimulated	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
mechanism	NN	O	O
for	NN	O	O
IL-10	NN	O	B-protein
suppression	NN	O	O
of	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

IL-4	NN	O	B-protein
,	NN	O	O
another	NN	O	O
cytokine	NN	O	B-protein
that	NN	O	O
inhibits	NN	O	O
cytokine	NN	O	B-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
shows	NN	O	O
little	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
LPS-induced	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Further	NN	O	O
examination	NN	O	O
reveals	NN	O	O
that	NN	O	O
,	NN	O	O
unlike	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
enhances	NN	O	O
mRNA	NN	O	B-RNA
degradation	NN	O	O
and	NN	O	O
does	NN	O	O
not	NN	O	O
suppress	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
inhibit	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
by	NN	O	O
different	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

-DOCSTART-	O

LMP-1	NN	O	B-protein
activates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
targeting	NN	O	O
the	NN	O	O
inhibitory	NN	O	B-protein
molecule	NN	O	I-protein
I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

LMP-1	NN	O	B-protein
,	NN	O	O
an	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
expressed	NN	O	O
during	NN	O	O
latent	NN	O	O
infection	NN	O	O
,	NN	O	O
has	NN	O	O
oncogenic	NN	O	O
properties	NN	O	O
,	NN	O	O
as	NN	O	O
judged	NN	O	O
from	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
transform	NN	O	O
B	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
rodent	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

LMP-1	NN	O	B-protein
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
bcl2	NN	O	B-DNA
,	NN	O	O
an	NN	O	O
oncogene	NN	O	B-DNA
which	NN	O	O
protects	NN	O	O
cells	NN	O	O
from	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
of	NN	O	O
genes	NN	O	O
encoding	NN	O	O
other	NN	O	O
proteins	NN	O	O
involved	NN	O	O
in	NN	O	O
cell	NN	O	O
regulation	NN	O	O
and	NN	O	O
growth	NN	O	O
control	NN	O	O
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
LMP-1	NN	O	B-protein
upregulates	NN	O	O
these	NN	O	O
proteins	NN	O	O
is	NN	O	O
unknown	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
is	NN	O	O
plausible	NN	O	O
that	NN	O	O
LMP-1	NN	O	B-protein
modifies	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
that	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
ultimately	NN	O	O
regulate	NN	O	O
transcription	NN	O	O
of	NN	O	O
oncogenic	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
controlling	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
involved	NN	O	O
in	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
growth	NN	O	O
control	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
LMP-1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
regulating	NN	O	O
this	NN	O	O
activation	NN	O	O
remains	NN	O	O
unknown	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
increased	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
functional	NN	O	O
activity	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
B-lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
stably	NN	O	O
or	NN	O	O
transiently	NN	O	O
expressing	NN	O	O
LMP-1	NN	O	B-protein
.	NN	O	O

I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
is	NN	O	O
selectively	NN	O	O
modified	NN	O	O
in	NN	O	O
LMP-1-expressing	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
phosphorylated	NN	O	O
form	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
increased	NN	O	O
protein	NN	O	O
turnover-degradation	NN	O	O
correlate	NN	O	O
with	NN	O	O
increased	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

This	NN	O	O
results	NN	O	O
in	NN	O	O
increased	NN	O	O
transcription	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B-dependent-genes	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
those	NN	O	O
encoding	NN	O	O
p105	NN	O	B-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
MAD3	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
LMP-1	NN	O	B-protein
activates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
by	NN	O	O
targeting	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
the	NN	O	O
pathways	NN	O	O
activated	NN	O	O
by	NN	O	O
LMP-1	NN	O	B-protein
to	NN	O	O
result	NN	O	O
in	NN	O	O
posttranslational	NN	O	O
modifications	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
will	NN	O	O
aid	NN	O	O
in	NN	O	O
determining	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
virus-host	NN	O	O
cell	NN	O	O
protein	NN	O	O
interaction	NN	O	O
in	NN	O	O
Epstein-Barr	NN	O	O
virus-mediated	NN	O	O
oncogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
TR2	NN	O	I-DNA
orphan	NN	O	I-DNA
receptor	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
simian	NN	O	B-DNA
virus	NN	O	I-DNA
40	NN	O	I-DNA
major	NN	O	I-DNA
late	NN	O	I-DNA
promoter	NN	O	I-DNA
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
J	NN	O	O
Biol	NN	O	O
Chem	NN	O	O
1995	NN	O	O
Nov	NN	O	O
3	NN	O	O
;	NN	O	O
270	NN	O	O
(	NN	O	O
44	NN	O	O
)	NN	O	O
:	NN	O	O
26721	NN	O	O
]	NN	O	O

A	NN	O	O
DNA	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
TR2RE-SV40	NN	O	B-DNA
)	NN	O	O
for	NN	O	O
the	NN	O	O
TR2	NN	O	B-protein
orphan	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
steroid-thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
,	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
simian	NN	O	B-DNA
virus	NN	O	I-DNA
40	NN	O	I-DNA
(	NN	O	I-DNA
SV40	NN	O	I-DNA
)	NN	O	I-DNA
+55	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
nucleotide	NN	O	B-DNA
numbers	NN	O	I-DNA
368-389	NN	O	I-DNA
,	NN	O	O
5'-GTTAAGGTTCGTAGGTCATGGA-3	NN	O	O
'	NN	O	O
)	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
using	NN	O	O
in	NN	O	O
vitro	NN	O	O
translated	NN	O	O
TR2	NN	O	B-protein
orphan	NN	O	I-protein
receptor	NN	O	I-protein
with	NN	O	O
a	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
67	NN	O	B-protein
kilodaltons	NN	O	I-protein
,	NN	O	O
showed	NN	O	O
a	NN	O	O
specific	NN	O	O
binding	NN	O	O
with	NN	O	O
high	NN	O	O
affinity	NN	O	O
(	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
=	NN	O	O
9	NN	O	O
nM	NN	O	O
)	NN	O	O
for	NN	O	O
this	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
.	NN	O	O

DNA-swap	NN	O	O
experiments	NN	O	O
using	NN	O	O
chloramphenicol	NN	O	B-protein
acetyl-transferase	NN	O	I-protein
assay	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
androgen	NN	O	O
can	NN	O	O
suppress	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
of	NN	O	O
SV40	NN	O	B-DNA
early	NN	O	I-DNA
promoter	NN	O	I-DNA
via	NN	O	O
the	NN	O	O
interaction	NN	O	O
between	NN	O	O
this	NN	O	O
TR2RE-SV40	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
receptor	NN	O	I-protein
AR/TR2/AR	NN	O	I-protein
with	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
TR2	NN	O	B-protein
orphan	NN	O	I-protein
receptor	NN	O	I-protein
flanked	NN	O	O
by	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
and	NN	O	I-protein
androgen-binding	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
the	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
this	NN	O	O
TR2RE-SV40	NN	O	B-DNA
can	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
repressor	NN	O	O
to	NN	O	O
suppress	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
of	NN	O	O
both	NN	O	O
SV40	NN	O	B-DNA
early	NN	O	I-DNA
and	NN	O	I-DNA
late	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
the	NN	O	O
TR2RE-SV40	NN	O	B-DNA
may	NN	O	O
represent	NN	O	O
the	NN	O	O
first	NN	O	O
identified	NN	O	O
natural	NN	O	O
DNA	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
TR2	NN	O	B-protein
orphan	NN	O	I-protein
receptor	NN	O	I-protein
that	NN	O	O
may	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
repressor	NN	O	O
for	NN	O	O
the	NN	O	O
SV40	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Detection	NN	O	O
of	NN	O	O
the	NN	O	O
chromosome	NN	O	B-RNA
16	NN	O	I-RNA
CBF	NN	O	I-RNA
beta-MYH11	NN	O	I-RNA
fusion	NN	O	I-RNA
transcript	NN	O	I-RNA
in	NN	O	O
myelomonocytic	NN	O	O
leukemias	NN	O	O
.	NN	O	O

Karyotypic	NN	O	O
detection	NN	O	O
of	NN	O	O
chromosomal	NN	O	O
16	NN	O	O
abnormalities	NN	O	O
classically	NN	O	O
associated	NN	O	O
with	NN	O	O
AML	NN	O	O
M4Eo	NN	O	O
can	NN	O	O
be	NN	O	O
difficult	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
inv	NN	O	O
(	NN	O	O
16	NN	O	O
)	NN	O	O
(	NN	O	O
p13q22	NN	O	O
)	NN	O	O
,	NN	O	O
CBF	NN	O	B-DNA
beta	NN	O	I-DNA
and	NN	O	O
MYH11	NN	O	B-DNA
,	NN	O	O
has	NN	O	O
allowed	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
fusion	NN	O	B-RNA
transcripts	NN	O	I-RNA
by	NN	O	O
reverse-transcriptase	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
CBF	NN	O	B-RNA
beta-MYH11	NN	O	I-RNA
fusion	NN	O	I-RNA
transcripts	NN	O	I-RNA
by	NN	O	O
RT-PCR	NN	O	O
in	NN	O	O
myelomonocytic	NN	O	O
leukemias	NN	O	O
,	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
eosinophilia	NN	O	O
,	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
their	NN	O	O
presence	NN	O	O
correlates	NN	O	O
with	NN	O	O
morphology	NN	O	O
.	NN	O	O

Fifty-three	NN	O	O
cases	NN	O	O
(	NN	O	O
11	NN	O	O
AML	NN	O	O
M4Eo	NN	O	O
;	NN	O	O
1	NN	O	O
AML	NN	O	O
M4	NN	O	O
with	NN	O	O
atypical	NN	O	B-cell_type
abnormal	NN	O	I-cell_type
eosinophils	NN	O	I-cell_type
(	NN	O	O
AML	NN	O	O
M4	NN	O	O
``	NN	O	O
Eo	NN	O	O
''	NN	O	O
)	NN	O	O
;	NN	O	O
29	NN	O	O
AML	NN	O	O
M4	NN	O	O
;	NN	O	O
8	NN	O	O
AML	NN	O	O
M5	NN	O	O
;	NN	O	O
3	NN	O	O
CMML	NN	O	O
;	NN	O	O
and	NN	O	O
1	NN	O	O
AML	NN	O	O
M2	NN	O	O
with	NN	O	O
eosinophilia	NN	O	O
)	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

All	NN	O	O
11	NN	O	O
typical	NN	O	O
AML	NN	O	O
M4Eo	NN	O	O
were	NN	O	O
CBF	NN	O	B-DNA
beta	NN	O	I-DNA
-MYH11	NN	O	B-DNA
positive	NN	O	O
.	NN	O	O

The	NN	O	O
single	NN	O	O
case	NN	O	O
of	NN	O	O
AML	NN	O	O
M4	NN	O	O
with	NN	O	O
distinctive	NN	O	O
eosinophil	NN	O	O
abnormalities	NN	O	O
was	NN	O	O
negative	NN	O	O
by	NN	O	O
karyotype	NN	O	O
,	NN	O	O
RT-PCR	NN	O	O
and	NN	O	O
fluorescent	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
.	NN	O	O

Three	NN	O	O
of	NN	O	O
29	NN	O	O
(	NN	O	O
10	NN	O	O
%	NN	O	O
)	NN	O	O
AML	NN	O	O
M4	NN	O	O
without	NN	O	O
abnormal	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
were	NN	O	O
CBF	NN	O	B-DNA
beta	NN	O	I-DNA
-MYH11	NN	O	B-DNA
positive	NN	O	O
,	NN	O	O
1	NN	O	O
of	NN	O	O
which	NN	O	O
did	NN	O	O
not	NN	O	O
show	NN	O	O
any	NN	O	O
apparent	NN	O	O
chromosome	NN	O	B-DNA
16	NN	O	I-DNA
abnormalities	NN	O	O
by	NN	O	O
classical	NN	O	O
metaphase	NN	O	O
analysis	NN	O	O
(	NN	O	O
2	NN	O	O
not	NN	O	O
tested	NN	O	O
)	NN	O	O
.	NN	O	O

Both	NN	O	O
cases	NN	O	O
tested	NN	O	O
also	NN	O	O
showed	NN	O	O
MYH11	NN	O	B-DNA
genomic	NN	O	O
rearrangement	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
leukemias	NN	O	O
were	NN	O	O
RT-PCR	NN	O	O
positive	NN	O	O
.	NN	O	O

Follow-up	NN	O	O
of	NN	O	O
three	NN	O	O
patient	NN	O	O
showed	NN	O	O
residual	NN	O	O
positivity	NN	O	O
in	NN	O	O
apparent	NN	O	O
complete	NN	O	O
remission	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
CBF	NN	O	B-RNA
beta-MYH11	NN	O	I-RNA
fusion	NN	O	I-RNA
transcripts	NN	O	I-RNA
occur	NN	O	O
not	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
vast	NN	O	O
majority	NN	O	O
of	NN	O	O
typical	NN	O	O
AML	NN	O	O
M4Eo	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
approximately	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
AML	NN	O	O
M4	NN	O	O
without	NN	O	O
eosinophilic	NN	O	O
abnormalities	NN	O	O
,	NN	O	O
a	NN	O	O
much	NN	O	O
higher	NN	O	O
incidence	NN	O	O
than	NN	O	O
the	NN	O	O
sporadic	NN	O	O
reports	NN	O	O
of	NN	O	O
chromosome	NN	O	B-DNA
16	NN	O	I-DNA
abnormalities	NN	O	O
in	NN	O	O
AML	NN	O	O
M4	NN	O	O
would	NN	O	O
suggest	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
with	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
CBF	NN	O	B-DNA
beta	NN	O	I-DNA
-	NN	O	O
MYH11	NN	O	B-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
apparent	NN	O	O
chromosome	NN	O	B-DNA
16	NN	O	I-DNA
abnormalities	NN	O	O
by	NN	O	O
classical	NN	O	O
banding	NN	O	O
techniques	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
additional	NN	O	O
screening	NN	O	O
by	NN	O	O
either	NN	O	O
RT-PCR	NN	O	O
or	NN	O	O
FISH	NN	O	O
should	NN	O	O
be	NN	O	O
performed	NN	O	O
in	NN	O	O
all	NN	O	O
AML	NN	O	O
M4	NN	O	O
,	NN	O	O
regardless	NN	O	O
of	NN	O	O
morphologic	NN	O	O
features	NN	O	O
,	NN	O	O
to	NN	O	O
allow	NN	O	O
accurate	NN	O	O
evaluation	NN	O	O
of	NN	O	O
the	NN	O	O
prognostic	NN	O	O
importance	NN	O	O
of	NN	O	O
this	NN	O	O
fusion	NN	O	B-RNA
transcript	NN	O	I-RNA
.	NN	O	O

-DOCSTART-	O

HIV-1	NN	O	B-protein
Tat	NN	O	I-protein
potentiates	NN	O	O
TNF	NN	O	B-protein
-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
cytotoxicity	NN	O	O
by	NN	O	O
altering	NN	O	O
the	NN	O	O
cellular	NN	O	O
redox	NN	O	O
state	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
human	NN	O	B-protein
immunodeficiency	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
HIV-1	NN	O	I-protein
)	NN	O	I-protein
Tat	NN	O	I-protein
protein	NN	O	I-protein
amplifies	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
cytokine	NN	O	O
that	NN	O	O
stimulates	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

In	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
tat	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
HeLa-tat	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Tat	NN	O	B-protein
protein	NN	O	I-protein
enhanced	NN	O	O
both	NN	O	O
TNF	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
TNF	NN	O	B-protein
-mediated	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
potentiation	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
effects	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
Jurkat	NN	O	O
T	NN	O	O
cells	NN	O	O
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
treated	NN	O	O
with	NN	O	O
soluble	NN	O	O
Tat	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

TNF	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
cytotoxicity	NN	O	O
involves	NN	O	O
the	NN	O	O
intracellular	NN	O	O
formation	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
Tat	NN	O	B-protein
-mediated	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
cellular	NN	O	O
redox	NN	O	O
state	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
HIV-1	NN	O	B-protein
Tat	NN	O	I-protein
suppressed	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Mn-dependent	NN	O	B-protein
superoxide	NN	O	I-protein
dismutase	NN	O	I-protein
(	NN	O	O
Mn-SOD	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
mitochondrial	NN	O	B-protein
enzyme	NN	O	I-protein
that	NN	O	O
is	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	O
defense	NN	O	O
system	NN	O	O
against	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Mn-SOD	NN	O	B-protein
RNA	NN	O	O
protein	NN	O	O
levels	NN	O	O
and	NN	O	O
activity	NN	O	O
were	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Tat	NN	O	B-protein
.	NN	O	O

Decreased	NN	O	O
Mn-SOD	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
decreased	NN	O	O
levels	NN	O	O
of	NN	O	O
glutathione	NN	O	O
and	NN	O	O
a	NN	O	O
lower	NN	O	O
ratio	NN	O	O
of	NN	O	O
reduced	NN	O	O
:	NN	O	O
oxidized	NN	O	O
glutathione	NN	O	O
.	NN	O	O

A	NN	O	O
truncated	NN	O	O
Tat	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
Tat1-72	NN	O	B-protein
)	NN	O	O
,	NN	O	O
known	NN	O	O
to	NN	O	O
transactivate	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
no	NN	O	O
longer	NN	O	O
affected	NN	O	O
Mn-SOD	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
the	NN	O	O
cellular	NN	O	O
redox	NN	O	O
state	NN	O	O
or	NN	O	O
TNF	NN	O	B-protein
-mediated	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
our	NN	O	O
experiments	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
HIV-1	NN	O	B-protein
Tat	NN	O	I-protein
is	NN	O	O
required	NN	O	O
to	NN	O	O
suppress	NN	O	O
Mn-SOD	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
to	NN	O	O
induce	NN	O	O
pro-oxidative	NN	O	O
conditions	NN	O	O
reflected	NN	O	O
by	NN	O	O
a	NN	O	O
drop	NN	O	O
in	NN	O	O
reduced	NN	O	O
glutathione	NN	O	O
(	NN	O	O
GSH	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
GSH	NN	O	O
:	NN	O	O
oxidized	NN	O	O
GSH	NN	O	O
(	NN	O	O
GSSG	NN	O	O
)	NN	O	O
ratio	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

The	NN	O	O
use	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
of	NN	O	O
childhood	NN	O	O
.	NN	O	O

Molecular	NN	O	O
,	NN	O	O
cellular	NN	O	O
,	NN	O	O
and	NN	O	O
clinical	NN	O	O
considerations	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
have	NN	O	O
been	NN	O	O
included	NN	O	O
in	NN	O	O
almost	NN	O	O
all	NN	O	O
treatment	NN	O	O
regimens	NN	O	O
for	NN	O	O
childhood	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
for	NN	O	O
decades	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
optimal	NN	O	O
agents	NN	O	O
,	NN	O	O
doses	NN	O	O
,	NN	O	O
and/or	NN	O	O
schedules	NN	O	O
have	NN	O	O
yet	NN	O	O
to	NN	O	O
be	NN	O	O
defined	NN	O	O
despite	NN	O	O
extensive	NN	O	O
clinical	NN	O	O
application	NN	O	O
.	NN	O	O

New	NN	O	O
data	NN	O	O
on	NN	O	O
the	NN	O	O
pharmacokinetics	NN	O	O
,	NN	O	O
pharmacodynamics	NN	O	O
,	NN	O	O
and	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
have	NN	O	O
suggested	NN	O	O
alternative	NN	O	O
approaches	NN	O	O
in	NN	O	O
ALL	NN	O	O
.	NN	O	O

These	NN	O	O
suggest	NN	O	O
that	NN	O	O
prolonged	NN	O	O
,	NN	O	O
i.e.	NN	O	O
28	NN	O	O
day	NN	O	O
,	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
may	NN	O	O
be	NN	O	O
unnecessary	NN	O	O
as	NN	O	O
exposure	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
induces	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Dexamethasone	NN	O	O
may	NN	O	O
be	NN	O	O
superior	NN	O	O
to	NN	O	O
prednisone	NN	O	O
in	NN	O	O
conventional	NN	O	O
equi-effective	NN	O	O
doses	NN	O	O
.	NN	O	O

Blast	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
correlates	NN	O	O
closely	NN	O	O
with	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
other	NN	O	O
,	NN	O	O
putatively	NN	O	O
non-cross-resisting	NN	O	O
agents	NN	O	O
and	NN	O	O
with	NN	O	O
outcome	NN	O	O
after	NN	O	O
multi-agent	NN	O	O
therapy	NN	O	O
,	NN	O	O
suggesting	NN	O	O
overlapping	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
action	NN	O	O
,	NN	O	O
and	NN	O	O
focusing	NN	O	O
attention	NN	O	O
on	NN	O	O
the	NN	O	O
determinants	NN	O	O
of	NN	O	O
the	NN	O	O
threshold	NN	O	O
for	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Increasing	NN	O	O
success	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
childhood	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
has	NN	O	O
led	NN	O	O
to	NN	O	O
increasing	NN	O	O
awareness	NN	O	O
of	NN	O	O
avascular	NN	O	O
necrosis	NN	O	O
of	NN	O	O
bone	NN	O	O
as	NN	O	O
a	NN	O	O
potentially	NN	O	O
disabling	NN	O	O
sequela	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
,	NN	O	O
especially	NN	O	O
in	NN	O	O
adolescent	NN	O	O
and	NN	O	O
young	NN	O	O
adult	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
2	NN	O	I-protein
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
latent	NN	O	B-DNA
membrane	NN	O	I-DNA
protein	NN	O	I-DNA
1	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
J	NN	O	B-protein
kappa	NN	O	I-protein
and	NN	O	O
PU.1	NN	O	B-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
EBV	NN	O	I-DNA
)	NN	O	I-DNA
latent	NN	O	I-DNA
membrane	NN	O	I-DNA
protein	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	I-DNA
LMP-1	NN	O	I-DNA
)	NN	O	I-DNA
oncogene	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	I-protein
EBNA-2	NN	O	I-protein
)	NN	O	I-protein
transactivator	NN	O	I-protein
.	NN	O	O

EBNA-2	NN	O	B-protein
is	NN	O	O
known	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
DNA-binding	NN	O	I-protein
protein	NN	O	I-protein
J	NN	O	B-protein
kappa	NN	O	I-protein
and	NN	O	O
is	NN	O	O
recruited	NN	O	O
to	NN	O	O
promoters	NN	O	B-DNA
containing	NN	O	O
the	NN	O	O
GTGGGAA	NN	O	O
J	NN	O	B-DNA
kappa	NN	O	I-DNA
recognition	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
minimal	NN	O	B-DNA
EBNA-2-responsive	NN	O	I-DNA
LMP-1	NN	O	I-DNA
promoter	NN	O	I-DNA
includes	NN	O	O
one	NN	O	O
J	NN	O	B-DNA
kappa-binding	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
we	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
mutation	NN	O	O
of	NN	O	O
that	NN	O	O
site	NN	O	O
,	NN	O	O
such	NN	O	O
that	NN	O	O
J	NN	O	B-protein
kappa	NN	O	I-protein
can	NN	O	O
not	NN	O	O
bind	NN	O	O
,	NN	O	O
reduces	NN	O	O
EBNA-2	NN	O	B-protein
responsiveness	NN	O	O
by	NN	O	O
60	NN	O	O
%	NN	O	O
.	NN	O	O

To	NN	O	O
identify	NN	O	O
other	NN	O	O
factors	NN	O	O
which	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
LMP-1	NN	O	B-DNA
EBNA-2	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
E2RE	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
a	NN	O	O
-236/-145	NN	O	B-DNA
minimal	NN	O	I-DNA
E2RE	NN	O	I-DNA
was	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
probe	NN	O	O
in	NN	O	O
an	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

The	NN	O	O
previously	NN	O	O
characterized	NN	O	O
factors	NN	O	O
J	NN	O	B-protein
kappa	NN	O	I-protein
,	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
AML1	NN	O	B-protein
bind	NN	O	O
to	NN	O	O
the	NN	O	O
LMP-1	NN	O	B-DNA
E2RE	NN	O	I-DNA
,	NN	O	O
along	NN	O	O
with	NN	O	O
six	NN	O	O
other	NN	O	O
unidentified	NN	O	O
factors	NN	O	O
(	NN	O	O
LBF2	NN	O	B-protein
to	NN	O	O
LBF7	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Binding	NN	O	O
sites	NN	O	O
were	NN	O	O
mapped	NN	O	O
for	NN	O	O
each	NN	O	O
factor	NN	O	O
.	NN	O	O

LBF4	NN	O	B-protein
is	NN	O	O
B-	NN	O	O
and	NN	O	O
T-cell	NN	O	O
specific	NN	O	O
and	NN	O	O
recognizes	NN	O	O
the	NN	O	O
PU.1	NN	O	B-DNA
GGAA	NN	O	I-DNA
core	NN	O	I-DNA
sequence	NN	O	I-DNA
as	NN	O	O
shown	NN	O	O
by	NN	O	O
methylation	NN	O	O
interference	NN	O	O
.	NN	O	O

LBF4	NN	O	B-protein
has	NN	O	O
a	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
105	NN	O	O
kDa	NN	O	O
and	NN	O	O
is	NN	O	O
probably	NN	O	O
unrelated	NN	O	O
to	NN	O	O
PU.1	NN	O	B-protein
.	NN	O	O

LBF2	NN	O	B-protein
was	NN	O	O
found	NN	O	O
only	NN	O	O
in	NN	O	O
epithelial	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
LBF3	NN	O	B-protein
,	NN	O	O
LBF5	NN	O	B-protein
,	NN	O	O
LBF6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
LBF7	NN	O	B-protein
were	NN	O	O
not	NN	O	O
cell	NN	O	O
type	NN	O	O
specific	NN	O	O
.	NN	O	O

Mutations	NN	O	O
of	NN	O	O
the	NN	O	O
AML1-	NN	O	B-DNA
or	NN	O	I-DNA
LBF4-binding	NN	O	I-DNA
sites	NN	O	I-DNA
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
EBNA-2	NN	O	B-protein
transactivation	NN	O	O
,	NN	O	O
whereas	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
PU.1-binding	NN	O	B-DNA
site	NN	O	I-DNA
completely	NN	O	O
eliminated	NN	O	O
EBNA-2	NN	O	B-protein
responses	NN	O	O
.	NN	O	O

A	NN	O	O
gst-EBNA-2	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
specifically	NN	O	O
depleted	NN	O	O
PU.1	NN	O	B-protein
from	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
and	NN	O	O
bound	NN	O	O
in	NN	O	O
vitro	NN	O	O
translated	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
providing	NN	O	O
biochemical	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
direct	NN	O	O
EBNA-2	NN	O	B-protein
-PU.1	NN	O	B-protein
interaction	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
EBNA-2	NN	O	B-protein
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
LMP-1	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
interaction	NN	O	O
with	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
distinct	NN	O	O
sequence-specific	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
J	NN	O	B-protein
kappa	NN	O	I-protein
and	NN	O	O
PU.1	NN	O	B-protein
.	NN	O	O

LBF3	NN	O	B-protein
,	NN	O	O
LBF5	NN	O	B-protein
,	NN	O	O
LBF6	NN	O	B-protein
,	NN	O	O
or	NN	O	O
LBF7	NN	O	B-protein
may	NN	O	O
also	NN	O	O
be	NN	O	O
involved	NN	O	O
,	NN	O	O
since	NN	O	O
their	NN	O	O
binding	NN	O	O
sites	NN	O	O
also	NN	O	O
contribute	NN	O	O
to	NN	O	O
EBNA-2	NN	O	B-protein
responsiveness	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lymphocyte	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
:	NN	O	O
predictor	NN	O	O
of	NN	O	O
sertraline	NN	O	O
response	NN	O	O
in	NN	O	O
adolescent	NN	O	O
major	NN	O	O
depressive	NN	O	O
disorder	NN	O	O
(	NN	O	O
MDD	NN	O	O
)	NN	O	O
.	NN	O	O

Major	NN	O	O
depressive	NN	O	O
disorder	NN	O	O
(	NN	O	O
MDD	NN	O	O
)	NN	O	O
in	NN	O	O
adolescents	NN	O	O
demonstrates	NN	O	O
resistance	NN	O	O
to	NN	O	O
tricyclic	NN	O	O
antidepressants	NN	O	O
and	NN	O	O
absence	NN	O	O
of	NN	O	O
hypercortisolemia	NN	O	O
.	NN	O	O

The	NN	O	O
efficacy	NN	O	O
of	NN	O	O
serotonin	NN	O	O
reuptake	NN	O	O
inhibitors	NN	O	O
(	NN	O	O
SRIs	NN	O	O
)	NN	O	O
is	NN	O	O
uncertain	NN	O	O
,	NN	O	O
and	NN	O	O
response	NN	O	O
predictors	NN	O	O
are	NN	O	O
unavailable	NN	O	O
.	NN	O	O

Abnormal	NN	O	O
fast	NN	O	O
feedback	NN	O	O
and	NN	O	O
negative	NN	O	O
feedback	NN	O	O
of	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
axis	NN	O	O
implicates	NN	O	O
a	NN	O	O
dampened	NN	O	O
limbic-hippocampal	NN	O	B-protein
glucocorticoid	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
GCII	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
lymphocyte	NN	O	B-protein
GCII	NN	O	I-protein
is	NN	O	O
altered	NN	O	O
in	NN	O	O
adolescent	NN	O	O
MDD	NN	O	O
and	NN	O	O
could	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
marker	NN	O	O
for	NN	O	O
response	NN	O	O
to	NN	O	O
SRIs	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
open-label	NN	O	O
study	NN	O	O
,	NN	O	O
adolescents	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
20	NN	O	O
)	NN	O	O
meeting	NN	O	O
DSM-III-R	NN	O	O
criteria	NN	O	O
for	NN	O	O
MDD	NN	O	O
showed	NN	O	O
baseline	NN	O	O
lymphocyte	NN	O	B-protein
GCII	NN	O	I-protein
sites	NN	O	I-protein
per	NN	O	O
cell	NN	O	O
(	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
values	NN	O	O
of	NN	O	O
793	NN	O	O
+/-	NN	O	O
106	NN	O	O
versus	NN	O	O
2	NN	O	O
,	NN	O	O
563	NN	O	O
+/-	NN	O	O
499	NN	O	O
(	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
)	NN	O	O
for	NN	O	O
matched	NN	O	O
controls	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
18	NN	O	O
)	NN	O	O
(	NN	O	O
t	NN	O	O
=	NN	O	O
3.5	NN	O	O
;	NN	O	O
df	NN	O	O
=	NN	O	O
36	NN	O	O
;	NN	O	O
p	NN	O	O
<	NN	O	O
.001	NN	O	O
)	NN	O	O
.	NN	O	O

GCII	NN	O	B-protein
was	NN	O	O
bimodally	NN	O	O
distributed	NN	O	O
,	NN	O	O
with	NN	O	O
SRI	NN	O	O
responders	NN	O	O
differing	NN	O	O
from	NN	O	O
nonresponders	NN	O	O
(	NN	O	O
t	NN	O	O
=	NN	O	O
3.9	NN	O	O
;	NN	O	O
df	NN	O	O
=	NN	O	O
14	NN	O	O
;	NN	O	O
p	NN	O	O
<	NN	O	O
.001	NN	O	O
)	NN	O	O
.	NN	O	O

GCII	NN	O	B-protein
accurately	NN	O	O
classified	NN	O	O
90	NN	O	O
percent	NN	O	O
of	NN	O	O
sertraline	NN	O	O
responders	NN	O	O
and	NN	O	O
80	NN	O	O
percent	NN	O	O
of	NN	O	O
nonresponders	NN	O	O
.	NN	O	O

Only	NN	O	O
SRI	NN	O	O
responders	NN	O	O
showed	NN	O	O
GCII	NN	O	B-protein
sites/cell	NN	O	O
upregulated	NN	O	O
after	NN	O	O
6	NN	O	O
weeks	NN	O	O
of	NN	O	O
treatment	NN	O	O
(	NN	O	O
t	NN	O	O
=	NN	O	O
2.1	NN	O	O
,	NN	O	O
df	NN	O	O
=	NN	O	O
10	NN	O	O
;	NN	O	O
p	NN	O	O
<	NN	O	O
.05	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
in	NN	O	O
cyclosporin	NN	O	B-DNA
A-sensitive	NN	O	I-DNA
tumor	NN	O	I-DNA
necrosis	NN	O	I-DNA
factor-alpha	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor-alpha	NN	O	I-DNA
(	NN	O	I-DNA
TNF	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
an	NN	O	O
immediate	NN	O	O
early	NN	O	O
gene	NN	O	O
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
in	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
without	NN	O	O
a	NN	O	O
requirement	NN	O	O
for	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Maximal	NN	O	O
induction	NN	O	O
of	NN	O	O
TNF	NN	O	B-RNA
alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
can	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
treatment	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
calcium	NN	O	O
ionophores	NN	O	O
alone	NN	O	O
,	NN	O	O
via	NN	O	O
a	NN	O	O
calcineurin	NN	O	B-protein
-dependent	NN	O	O
process	NN	O	O
that	NN	O	O
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
identified	NN	O	O
a	NN	O	O
promoter	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
kappa	NN	O	B-DNA
3	NN	O	I-DNA
,	NN	O	O
that	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
calcium-stimulated	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A-sensitive	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
kappa	NN	O	B-protein
3	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
contains	NN	O	O
NFATp	NN	O	B-protein
,	NN	O	O
a	NN	O	O
cyclosporin-sensitive	NN	O	B-protein
DNA-binding	NN	O	I-protein
protein	NN	O	I-protein
required	NN	O	O
for	NN	O	O
interleukin-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

NFATp	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
two	NN	O	O
sites	NN	O	O
within	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
3	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
occupancy	NN	O	O
of	NN	O	O
both	NN	O	O
sites	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	O
gene	NN	O	O
induction	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
3	NN	O	I-DNA
element	NN	O	I-DNA
has	NN	O	O
little	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
to	NN	O	O
other	NN	O	O
NFATp-binding	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
it	NN	O	O
appears	NN	O	O
to	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
cyclosporin-sensitive	NN	O	B-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
virtue	NN	O	O
of	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
bind	NN	O	O
NFATp	NN	O	B-protein
.	NN	O	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
in	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
and	NN	O	O
TNF	NN	O	B-DNA
alpha	NN	O	I-DNA
genes	NN	O	I-DNA
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
factor	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
coordinate	NN	O	O
induction	NN	O	O
of	NN	O	O
multiple	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
starting	NN	O	O
at	NN	O	O
the	NN	O	O
earliest	NN	O	O
stages	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
and	NN	O	O
specificity	NN	O	O
of	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
leukemia/B	NN	O	B-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
myeloid	NN	O	I-protein
cell	NN	O	I-protein
nuclear	NN	O	I-protein
differentiation	NN	O	I-protein
antigen	NN	O	I-protein
(	NN	O	O
MNDA	NN	O	B-protein
)	NN	O	O
was	NN	O	O
observed	NN	O	O
specifically	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
granulocyte-macrophage	NN	O	O
lineage	NN	O	O
in	NN	O	O
our	NN	O	O
earlier	NN	O	O
reports	NN	O	O
.	NN	O	O

The	NN	O	O
specificity	NN	O	O
of	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
for	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
granulocyte-macrophage	NN	O	O
lineage	NN	O	O
was	NN	O	O
reexamined	NN	O	O
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
established	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
Philadelphia	NN	O	O
chromosome-positive	NN	O	O
chronic	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Cell	NN	O	O
lines	NN	O	O
that	NN	O	O
expressed	NN	O	O
MNDA	NN	O	B-protein
exhibited	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
features	NN	O	O
and	NN	O	O
granulocyte	NN	O	O
or	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
could	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
while	NN	O	O
cell	NN	O	O
lines	NN	O	O
exhibiting	NN	O	O
properties	NN	O	O
of	NN	O	O
very	NN	O	O
early	NN	O	O
stage	NN	O	O
cells	NN	O	O
or	NN	O	O
multipotential	NN	O	B-cell_type
cells	NN	O	I-cell_type
did	NN	O	O
not	NN	O	O
express	NN	O	O
MNDA	NN	O	B-protein
.	NN	O	O

Cells	NN	O	O
originating	NN	O	O
from	NN	O	O
cases	NN	O	O
of	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
were	NN	O	O
negative	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
three	NN	O	O
lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
immortalized	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
)	NN	O	O
were	NN	O	O
weakly	NN	O	O
positive	NN	O	O
and	NN	O	O
MNDA	NN	O	B-protein
was	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
interferon-alpha	NN	O	B-protein
(	NN	O	O
IFN-alpha	NN	O	B-protein
)	NN	O	O
treatment	NN	O	O
.	NN	O	O

As	NN	O	O
we	NN	O	O
reported	NN	O	O
previously	NN	O	O
,	NN	O	O
MNDA	NN	O	B-RNA
mRNA	NN	O	I-RNA
level	NN	O	O
in	NN	O	O
adherent	NN	O	O
monocytes	NN	O	B-cell_type
is	NN	O	O
elevated	NN	O	O
by	NN	O	O
IFN-alpha	NN	O	B-protein
;	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
further	NN	O	O
assessed	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
in	NN	O	O
vitro	NN	O	O
monocyte-derived	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

Three	NN	O	O
additional	NN	O	O
agents	NN	O	O
(	NN	O	O
endotoxin	NN	O	O
,	NN	O	O
phytohemagglutinin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
)	NN	O	O
and	NN	O	O
other	NN	O	O
conditions	NN	O	O
that	NN	O	O
affect	NN	O	O
function	NN	O	O
,	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and/or	NN	O	O
growth	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
were	NN	O	O
examined	NN	O	O
for	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
alter	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
varied	NN	O	O
with	NN	O	O
the	NN	O	O
agent	NN	O	O
,	NN	O	O
cell	NN	O	O
type	NN	O	O
,	NN	O	O
and	NN	O	O
stage	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Changes	NN	O	O
in	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
occurred	NN	O	O
slowly	NN	O	O
(	NN	O	O
hours	NN	O	O
to	NN	O	O
days	NN	O	O
)	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
MNDA	NN	O	B-protein
could	NN	O	O
mediate	NN	O	O
changes	NN	O	O
realized	NN	O	O
over	NN	O	O
a	NN	O	O
long	NN	O	O
period	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
also	NN	O	O
reveal	NN	O	O
a	NN	O	O
discordance	NN	O	O
in	NN	O	O
certain	NN	O	O
MNDA	NN	O	B-cell_line
positive	NN	O	I-cell_line
cells	NN	O	I-cell_line
between	NN	O	O
steady-state	NN	O	O
levels	NN	O	O
or	NN	O	O
changes	NN	O	O
in	NN	O	O
levels	NN	O	O
of	NN	O	O
protein	NN	O	O
and	NN	O	O
mRNA	NN	O	B-RNA
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
occurs	NN	O	O
at	NN	O	O
more	NN	O	O
than	NN	O	O
one	NN	O	O
point	NN	O	O
.	NN	O	O

Changes	NN	O	O
in	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
opposing	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
/macrophages	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

DNA-binding	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	O
EBNA-2	NN	O	B-protein
)	NN	O	O
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
complex	NN	O	O
formation	NN	O	O
by	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
gene	NN	O	I-protein
promoter-binding	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
EBNA-2-positive	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
(	NN	O	I-protein
EBV	NN	O	I-protein
)	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	O
EBNA-2	NN	O	B-protein
)	NN	O	O
protein	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
immortalization	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
primary	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
EBV	NN	O	O
.	NN	O	O

EBNA-2	NN	O	B-protein
trans-activates	NN	O	O
cellular	NN	O	B-DNA
and	NN	O	I-DNA
viral	NN	O	I-DNA
genes	NN	O	I-DNA
like	NN	O	O
CD23	NN	O	B-DNA
,	NN	O	O
c-fgr	NN	O	B-DNA
,	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
LMP1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
terminal	NN	O	B-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
TP1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Trans-activation	NN	O	O
of	NN	O	O
the	NN	O	O
TP1	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
of	NN	O	O
the	NN	O	O
BamHI	NN	O	B-DNA
C	NN	O	I-DNA
promoter	NN	O	I-DNA
has	NN	O	O
already	NN	O	O
been	NN	O	O
investigated	NN	O	O
in	NN	O	O
detail	NN	O	O
and	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
via	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
and	NN	O	O
not	NN	O	O
by	NN	O	O
direct	NN	O	O
binding	NN	O	O
of	NN	O	O
EBNA-2	NN	O	B-protein
type	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	O
of	NN	O	O
EBV	NN	O	O
type	NN	O	O
1	NN	O	O
)	NN	O	O
to	NN	O	O
the	NN	O	O
DNA	NN	O	O
.	NN	O	O

EBNA-2	NN	O	B-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
trans-activate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
LMP	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
several	NN	O	O
cell	NN	O	O
lines	NN	O	O
.	NN	O	O

Various	NN	O	O
reports	NN	O	O
have	NN	O	O
delineated	NN	O	O
the	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
LMP	NN	O	B-DNA
promoter	NN	O	I-DNA
through	NN	O	O
which	NN	O	O
EBNA-2	NN	O	B-protein
mediates	NN	O	O
trans-activation	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
EBNA-2	NN	O	B-protein
also	NN	O	O
trans-activates	NN	O	O
the	NN	O	O
LMP	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
,	NN	O	O
we	NN	O	O
performed	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
gel	NN	O	O
retardation	NN	O	O
assays	NN	O	O
and	NN	O	O
competition	NN	O	O
experiments	NN	O	O
with	NN	O	O
LMP	NN	O	B-DNA
promoter	NN	O	I-DNA
fragments	NN	O	I-DNA
of	NN	O	O
different	NN	O	O
sizes	NN	O	O
.	NN	O	O

We	NN	O	O
determined	NN	O	O
that	NN	O	O
the	NN	O	O
protein-binding	NN	O	B-DNA
region	NN	O	I-DNA
on	NN	O	O
the	NN	O	O
LMP	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
within	NN	O	O
a	NN	O	O
42	NN	O	B-DNA
bp	NN	O	I-DNA
fragment	NN	O	I-DNA
encompassing	NN	O	O
nucleotides	NN	O	O
-135	NN	O	O
to	NN	O	O
-176	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
LMP	NN	O	B-DNA
transcriptional	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
fragments	NN	O	O
investigated	NN	O	O
indicated	NN	O	O
interaction	NN	O	O
of	NN	O	O
EBNA-2	NN	O	B-protein
with	NN	O	O
the	NN	O	O
DNA	NN	O	O
via	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
.	NN	O	O

No	NN	O	O
significant	NN	O	O
differences	NN	O	O
between	NN	O	O
EBNA-2	NN	O	B-protein
-positive	NN	O	O
and	NN	O	O
EBNA-2	NN	O	B-protein
-negative	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
could	NN	O	O
be	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
gel	NN	O	O
retardation	NN	O	O
assay	NN	O	O
under	NN	O	O
conditions	NN	O	O
that	NN	O	O
clearly	NN	O	O
showed	NN	O	O
binding	NN	O	O
of	NN	O	O
EBNA-2A	NN	O	B-protein
to	NN	O	O
the	NN	O	O
TP1	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
analysis	NN	O	O
of	NN	O	O
sucrose	NN	O	O
gradient	NN	O	O
fractions	NN	O	O
in	NN	O	O
the	NN	O	O
gel	NN	O	O
retardation	NN	O	O
assay	NN	O	O
provided	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
LMP	NN	O	B-protein
promoter-binding	NN	O	I-protein
proteins	NN	O	I-protein
form	NN	O	O
a	NN	O	O
complex	NN	O	O
of	NN	O	O
higher	NN	O	O
M	NN	O	O
(	NN	O	O
r	NN	O	O
)	NN	O	O
in	NN	O	O
EBNA-2-positive	NN	O	B-cell_type
cell	NN	O	I-cell_type
extracts	NN	O	O
.	NN	O	O

These	NN	O	O
complexes	NN	O	O
were	NN	O	O
destroyed	NN	O	O
by	NN	O	O
detergent	NN	O	O
.	NN	O	O

We	NN	O	O
deduce	NN	O	O
from	NN	O	O
these	NN	O	O
results	NN	O	O
that	NN	O	O
EBNA-2-positive	NN	O	B-cell_line
cells	NN	O	I-cell_line
might	NN	O	O
indeed	NN	O	O
contain	NN	O	O
specific	NN	O	B-protein
complexes	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
LMP	NN	O	B-DNA
promoter	NN	O	I-DNA
which	NN	O	O
are	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
too	NN	O	O
labile	NN	O	O
to	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
a	NN	O	O
standard	NN	O	O
gel	NN	O	O
retardation	NN	O	O
assay	NN	O	O
.	NN	O	O

-DOCSTART-	O

Simultaneous	NN	O	O
activation	NN	O	O
of	NN	O	O
Ig	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
synthesis	NN	O	O
and	NN	O	O
reduction	NN	O	O
of	NN	O	O
surface	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
IL-6	NN	O	B-protein
.	NN	O	O

Terminal	NN	O	O
differentiation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vivo	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
secretion	NN	O	O
of	NN	O	O
Ig	NN	O	B-protein
and	NN	O	O
extinction	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
IL-6	NN	O	B-protein
signaling	NN	O	O
leads	NN	O	O
to	NN	O	O
marked	NN	O	O
increases	NN	O	O
in	NN	O	O
the	NN	O	O
synthesis	NN	O	O
and	NN	O	O
secretion	NN	O	O
of	NN	O	O
Ig	NN	O	B-protein
in	NN	O	O
clonal	NN	O	B-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
newly	NN	O	O
isolated	NN	O	B-cell_type
polyclonal	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
IL-6-induced	NN	O	B-cell_line
cells	NN	O	I-cell_line
resemble	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
ultrastructure	NN	O	O
and	NN	O	O
in	NN	O	O
reduced	NN	O	O
expression	NN	O	O
of	NN	O	O
surface	NN	O	B-protein
MHC	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
.	NN	O	O

Enhanced	NN	O	O
Ig	NN	O	B-protein
synthesis	NN	O	O
is	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
coordinated	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
Ig	NN	O	B-DNA
genes	NN	O	I-DNA
without	NN	O	O
promoter	NN	O	O
or	NN	O	O
isotype	NN	O	O
specificity	NN	O	O
,	NN	O	O
and	NN	O	O
differential	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
encoding	NN	O	O
the	NN	O	O
secreted	NN	O	O
form	NN	O	O
of	NN	O	O
Ig	NN	O	B-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
saturable	NN	O	O
and	NN	O	O
subject	NN	O	O
to	NN	O	O
negative	NN	O	O
control	NN	O	O
when	NN	O	O
IL-6	NN	O	B-protein
stimulation	NN	O	O
is	NN	O	O
prolonged	NN	O	O
.	NN	O	O

Coordinate	NN	O	O
with	NN	O	O
temporal	NN	O	O
changes	NN	O	O
in	NN	O	O
Ig	NN	O	B-protein
synthesis	NN	O	O
,	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
and	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell-enriched	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Oct-2	NN	O	B-protein
are	NN	O	O
regulated	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
differentiation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
IL-6	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
recapitulates	NN	O	O
the	NN	O	O
hallmarks	NN	O	O
of	NN	O	O
terminal	NN	O	O
B	NN	O	O
differentiation	NN	O	O
in	NN	O	O
vivo	NN	O	O
;	NN	O	O
Oct-2	NN	O	B-protein
may	NN	O	O
have	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

-DOCSTART-	O

T-cell	NN	O	B-DNA
functional	NN	O	I-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-3	NN	O	I-DNA
proximal	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
interleukin-3	NN	O	I-DNA
(	NN	O	I-DNA
IL-3	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
expressed	NN	O	O
almost	NN	O	O
exclusively	NN	O	O
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Its	NN	O	O
expression	NN	O	O
is	NN	O	O
regulated	NN	O	O
at	NN	O	O
both	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
post-transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
treatment	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
phytohemaglutinin	NN	O	B-protein
(	NN	O	O
PHA	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
,	NN	O	O
PMA	NN	O	O
,	NN	O	O
activated	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
from	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
define	NN	O	O
the	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
required	NN	O	O
for	NN	O	O
transcription	NN	O	O
activation	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
reporter	NN	O	O
constructs	NN	O	O
containing	NN	O	O
different	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
gene	NN	O	I-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
and	NN	O	I-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
positive	NN	O	B-DNA
and	NN	O	I-DNA
negative	NN	O	I-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
between	NN	O	O
-173	NN	O	B-DNA
and	NN	O	I-DNA
-60	NN	O	I-DNA
contained	NN	O	O
the	NN	O	O
strongest	NN	O	O
activating	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
could	NN	O	O
bind	NN	O	O
to	NN	O	O
this	NN	O	O
positive	NN	O	O
activator	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
examined	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
CK-1/CK-2	NN	O	I-DNA
elements	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
many	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
found	NN	O	O
that	NN	O	O
they	NN	O	O
acted	NN	O	O
as	NN	O	O
a	NN	O	O
repressor	NN	O	O
of	NN	O	O
basal	NN	O	O
level	NN	O	O
expression	NN	O	O
when	NN	O	O
cloned	NN	O	O
upstream	NN	O	O
of	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
but	NN	O	O
were	NN	O	O
also	NN	O	O
inducible	NN	O	O
by	NN	O	O
PMA	NN	O	O
/PHA	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
Runt	NN	O	B-DNA
domain-encoding	NN	O	I-DNA
PEBP2	NN	O	I-DNA
alpha	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
thymic	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
PEBP2	NN	O	B-DNA
alpha	NN	O	I-DNA
A	NN	O	I-DNA
and	NN	O	I-DNA
PEBP2	NN	O	I-DNA
alpha	NN	O	I-DNA
B	NN	O	I-DNA
genes	NN	O	I-DNA
encode	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
a	NN	O	O
murine	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
PEBP2	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
implicated	NN	O	O
as	NN	O	O
a	NN	O	O
T-cell-specific	NN	O	B-protein
transcriptional	NN	O	I-protein
regulator	NN	O	I-protein
.	NN	O	O

These	NN	O	O
two	NN	O	O
related	NN	O	O
genes	NN	O	O
share	NN	O	O
the	NN	O	O
evolutionarily	NN	O	O
conserved	NN	O	O
region	NN	O	O
encoding	NN	O	O
the	NN	O	O
Runt	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

PEBP2	NN	O	B-protein
alpha	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
counterpart	NN	O	I-protein
of	NN	O	O
human	NN	O	B-protein
AML1	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
located	NN	O	O
at	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
of	NN	O	O
the	NN	O	O
8	NN	O	B-DNA
;	NN	O	I-DNA
21	NN	O	I-DNA
and	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
21	NN	O	I-DNA
chromosome	NN	O	I-DNA
translocations	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Northern	NN	O	O
(	NN	O	O
RNA	NN	O	O
)	NN	O	O
blots	NN	O	O
of	NN	O	O
various	NN	O	O
adult	NN	O	O
mouse	NN	O	O
tissues	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
genes	NN	O	O
were	NN	O	O
most	NN	O	O
prominent	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
transcripts	NN	O	O
of	NN	O	O
PEBP2	NN	O	B-protein
alpha	NN	O	I-protein
A	NN	O	I-protein
and	NN	O	O
mouse	NN	O	B-protein
AML1/PEBP2	NN	O	I-protein
alpha	NN	O	I-protein
B	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
thymuses	NN	O	O
from	NN	O	O
day	NN	O	O
16	NN	O	O
embryos	NN	O	O
and	NN	O	O
newborns	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
4-week-old	NN	O	O
adult	NN	O	O
mice	NN	O	O
,	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
persisted	NN	O	O
in	NN	O	O
peripheral	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
of	NN	O	O
adult	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
transcripts	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
all	NN	O	O
the	NN	O	O
CD4-	NN	O	B-cell_type
CD8-	NN	O	I-cell_type
,	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
,	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
CD8-	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
CD4-	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
cell	NN	O	I-cell_type
populations	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicated	NN	O	O
that	NN	O	O
both	NN	O	O
genes	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
throughout	NN	O	O
their	NN	O	O
development	NN	O	O
,	NN	O	O
supporting	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
PEBP2	NN	O	B-protein
is	NN	O	O
a	NN	O	O
T-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Transcripts	NN	O	O
of	NN	O	O
mouse	NN	O	B-protein
AML1/PEBP2	NN	O	I-protein
alpha	NN	O	I-protein
B	NN	O	I-protein
were	NN	O	O
also	NN	O	O
detected	NN	O	O
in	NN	O	O
day	NN	O	O
12	NN	O	O
fetal	NN	O	O
hematopoietic	NN	O	O
liver	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
newborn	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
implication	NN	O	O
of	NN	O	O
mouse	NN	O	O
AML1/PEBP2	NN	O	B-protein
alpha	NN	O	I-protein
B	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
other	NN	O	O
than	NN	O	O
those	NN	O	O
of	NN	O	O
T-cell	NN	O	B-cell_type
lineage	NN	O	I-cell_type
is	NN	O	O
discussed	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
myeloid	NN	O	O
leukemogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
on	NN	O	O
transcription	NN	O	O
factor	NN	O	O
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Glucocorticoids	NN	O	O
have	NN	O	O
an	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
inflammatory	NN	O	O
and	NN	O	O
immune	NN	O	O
responses	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
may	NN	O	O
be	NN	O	O
through	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
.	NN	O	O

The	NN	O	O
interaction	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
cAMP-responsive	NN	O	B-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
with	NN	O	O
DNA	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
was	NN	O	O
analyzed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

TNF-alpha	NN	O	O
,	NN	O	O
IL-1	NN	O	O
beta	NN	O	O
and	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
treatment	NN	O	O
increased	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF	NN	O	O
kappa	NN	O	O
B	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
up	NN	O	O
to	NN	O	O
200	NN	O	O
%	NN	O	O
but	NN	O	O
decreased	NN	O	O
CREB	NN	O	B-protein
binding	NN	O	O
(	NN	O	O
38	NN	O	O
%	NN	O	O
)	NN	O	O
over	NN	O	O
a	NN	O	O
60-min	NN	O	O
time	NN	O	O
course	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
produced	NN	O	O
a	NN	O	O
rapid	NN	O	O
and	NN	O	O
sustained	NN	O	O
increase	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
binding	NN	O	O
and	NN	O	O
a	NN	O	O
concomitant	NN	O	O
40-50	NN	O	O
%	NN	O	O
decrease	NN	O	O
in	NN	O	O
AP-1	NN	O	O
,	NN	O	O
NF	NN	O	O
kappa	NN	O	O
B	NN	O	O
,	NN	O	O
and	NN	O	O
CREB	NN	O	O
DNA	NN	O	O
binding	NN	O	O
that	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
combined	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
or	NN	O	O
PMA	NN	O	O
treatment	NN	O	O
.	NN	O	O

These	NN	O	O
latter	NN	O	O
effects	NN	O	O
were	NN	O	O
due	NN	O	O
to	NN	O	O
increases	NN	O	O
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
,	NN	O	O
not	NN	O	O
to	NN	O	O
reduced	NN	O	O
amounts	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
GR	NN	O	B-protein
within	NN	O	O
the	NN	O	O
nucleus	NN	O	O
interacts	NN	O	O
with	NN	O	O
cytokine-stimulated	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
by	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
cross	NN	O	O
coupling	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
molecular	NN	O	O
site	NN	O	O
of	NN	O	O
steroid	NN	O	O
action	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
complex	NN	O	I-protein
during	NN	O	O
restimulation	NN	O	O
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
T-cell	NN	O	O
anergy	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
through	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
TcR	NN	O	B-protein
)	NN	O	O
by	NN	O	O
high	NN	O	O
doses	NN	O	O
of	NN	O	O
specific	NN	O	O
peptide	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
long-lived	NN	O	O
state	NN	O	O
of	NN	O	O
nonresponsiveness	NN	O	O
that	NN	O	O
has	NN	O	O
been	NN	O	O
called	NN	O	O
anergy	NN	O	O
.	NN	O	O

During	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
anergy	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
phenotypically	NN	O	O
similar	NN	O	O
to	NN	O	O
cells	NN	O	O
responding	NN	O	O
to	NN	O	O
an	NN	O	O
immunogenic	NN	O	O
stimulus	NN	O	O
.	NN	O	O

The	NN	O	O
amount	NN	O	O
of	NN	O	O
TcR	NN	O	B-protein
at	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
is	NN	O	O
downmodulated	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
CD2	NN	O	B-protein
and	NN	O	I-protein
CD25	NN	O	I-protein
receptors	NN	O	I-protein
are	NN	O	O
increased	NN	O	O
.	NN	O	O

When	NN	O	O
restimulated	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
anergic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
fail	NN	O	O
to	NN	O	O
up-regulate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
in	NN	O	O
consequence	NN	O	O
do	NN	O	O
not	NN	O	O
produce	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
compared	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
various	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
bind	NN	O	O
to	NN	O	O
their	NN	O	O
appropriate	NN	O	O
site	NN	O	O
on	NN	O	O
DNA	NN	O	O
.	NN	O	O

Factors	NN	O	O
were	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
were	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
phase	NN	O	O
of	NN	O	O
anergy	NN	O	O
or	NN	O	O
were	NN	O	O
undergoing	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
pattern	NN	O	O
of	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
restimulated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
pattern	NN	O	O
that	NN	O	O
has	NN	O	O
previously	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
regulate	NN	O	O
T-cell-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
the	NN	O	O
beta	NN	O	O
chain	NN	O	O
of	NN	O	O
the	NN	O	O
TcR	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
measured	NN	O	O
binding	NN	O	O
to	NN	O	O
a	NN	O	O
TCF-1	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
the	NN	O	O
same	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
,	NN	O	I-cell_type
activated	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
anergized	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
inducible	NN	O	B-protein
factors	NN	O	I-protein
NK-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
beta	NN	O	B-protein
E2	NN	O	I-protein
,	NN	O	O
CD28RC	NN	O	B-protein
,	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
are	NN	O	O
not	NN	O	O
expressed	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
are	NN	O	O
twofold	NN	O	O
lower	NN	O	O
in	NN	O	O
anergized	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
activated	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
anergic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
approximately	NN	O	O
eightfold	NN	O	O
lower	NN	O	O
amounts	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	O
of	NN	O	O
inducible	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
regulates	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
elevated	NN	O	O
by	NN	O	O
a	NN	O	O
zinc	NN	O	O
containing	NN	O	O
trace	NN	O	O
element	NN	O	O
preparation	NN	O	O
.	NN	O	O

A	NN	O	O
trace	NN	O	O
element	NN	O	O
preparation	NN	O	O
(	NN	O	O
Beres	NN	O	O
Drops	NN	O	O
Plus	NN	O	O
,	NN	O	O
BDP	NN	O	O
)	NN	O	O
elevates	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
gcR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
isolated	NN	O	O
both	NN	O	O
from	NN	O	O
healthy	NN	O	O
blood	NN	O	O
donors	NN	O	O
and	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
patients	NN	O	O
.	NN	O	O

This	NN	O	O
enhancement	NN	O	O
by	NN	O	O
BDP	NN	O	O
was	NN	O	O
found	NN	O	O
either	NN	O	O
for	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
gcRs	NN	O	B-protein
or	NN	O	O
in	NN	O	O
experiments	NN	O	O
when	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
were	NN	O	O
stimulated	NN	O	O
by	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-6	NN	O	I-protein
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
significant	NN	O	O
effect	NN	O	O
of	NN	O	O
BDP	NN	O	O
on	NN	O	O
IL-1	NN	O	B-protein
and	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
-induced	NN	O	O
changes	NN	O	O
of	NN	O	O
gcRs	NN	O	B-protein
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
BDP	NN	O	O
was	NN	O	O
greatly	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Zn++	NN	O	O
ions	NN	O	O
in	NN	O	O
the	NN	O	O
preparation	NN	O	O
,	NN	O	O
since	NN	O	O
the	NN	O	O
augmenting	NN	O	O
effect	NN	O	O
was	NN	O	O
abolished	NN	O	O
if	NN	O	O
BDP	NN	O	O
did	NN	O	O
not	NN	O	O
contain	NN	O	O
zinc	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
DNA	NN	O	O
and	NN	O	O
steroid	NN	O	B-protein
binding	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
are	NN	O	O
not	NN	O	O
altered	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
treated	NN	O	O
CLL	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
investigate	NN	O	O
whether	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
could	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
increasing	NN	O	O
unresponsiveness	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
lymphatic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CLL	NN	O	O
)	NN	O	O
to	NN	O	O
combination	NN	O	O
chemotherapy	NN	O	O
.	NN	O	O

The	NN	O	O
receptor	NN	O	O
was	NN	O	O
tested	NN	O	O
immunocytochemically	NN	O	O
,	NN	O	O
in	NN	O	O
steroid	NN	O	O
binding	NN	O	O
assays	NN	O	O
,	NN	O	O
and	NN	O	O
by	NN	O	O
a	NN	O	O
mutation	NN	O	O
screening	NN	O	O
(	NN	O	O
denaturing	NN	O	O
gradient	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
receptor-cDNA	NN	O	B-protein
.	NN	O	O

The	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
staining	NN	O	O
and	NN	O	O
steroid	NN	O	O
binding	NN	O	O
test	NN	O	O
,	NN	O	O
varied	NN	O	O
considerably	NN	O	O
but	NN	O	O
showed	NN	O	O
no	NN	O	O
clear	NN	O	O
correlation	NN	O	O
to	NN	O	O
clinical	NN	O	O
response	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
highly	NN	O	O
sensitive	NN	O	O
mutation	NN	O	O
screening	NN	O	O
assay	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-	NN	O	B-protein
and	NN	O	I-protein
the	NN	O	I-protein
steroid-binding	NN	O	I-protein
region	NN	O	I-protein
,	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
treated	NN	O	O
patients	NN	O	O
revealed	NN	O	O
any	NN	O	O
mutation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
CLL	NN	O	O
patients	NN	O	O
tested	NN	O	O
is	NN	O	O
not	NN	O	O
altered	NN	O	O
in	NN	O	O
these	NN	O	O
domains	NN	O	O
.	NN	O	O

In	NN	O	O
one	NN	O	O
individual	NN	O	O
who	NN	O	O
had	NN	O	O
not	NN	O	O
been	NN	O	O
treated	NN	O	O
before	NN	O	O
analysis	NN	O	O
a	NN	O	O
silent	NN	O	O
mutation	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
one	NN	O	O
receptor	NN	O	O
allele	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
mechanisms	NN	O	O
other	NN	O	O
than	NN	O	O
altered	NN	O	O
ligand	NN	O	O
or	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
may	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
response	NN	O	O
to	NN	O	O
chemotherapy	NN	O	O
.	NN	O	O

This	NN	O	O
conclusion	NN	O	O
is	NN	O	O
discussed	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
corticoid	NN	O	O
resistance	NN	O	O
in	NN	O	O
mouse	NN	O	B-cell_line
and	NN	O	I-cell_line
human	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
culture	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
AP1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
intron	NN	O	I-DNA
enhancer	NN	O	I-DNA
binds	NN	O	O
inducible	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
contributes	NN	O	O
to	NN	O	O
expression	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
kappa	NN	O	O
immunoglobulin	NN	O	O
light	NN	O	O
chain	NN	O	O
gene	NN	O	O
requires	NN	O	O
developmental-	NN	O	O
and	NN	O	O
tissue-specific	NN	O	O
regulation	NN	O	O
by	NN	O	O
trans-acting	NN	O	B-protein
factors	NN	O	I-protein
which	NN	O	O
interact	NN	O	O
with	NN	O	O
two	NN	O	O
distinct	NN	O	O
enhancer	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
new	NN	O	O
protein-DNA	NN	O	O
interaction	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
intron	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
within	NN	O	O
the	NN	O	O
matrix-associated	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
J-C	NN	O	B-DNA
intron	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
greatly	NN	O	O
inducible	NN	O	O
in	NN	O	O
pre-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
and	NN	O	O
interleukin-1	NN	O	B-protein
but	NN	O	O
specific	NN	O	O
complexes	NN	O	O
are	NN	O	O
found	NN	O	O
at	NN	O	O
all	NN	O	O
stages	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
tested	NN	O	O
.	NN	O	O

The	NN	O	O
footprinted	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
is	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
AP1	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
protein	NN	O	O
components	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
are	NN	O	O
specifically	NN	O	O
competed	NN	O	O
by	NN	O	O
an	NN	O	O
AP1	NN	O	B-DNA
consensus	NN	O	I-DNA
motif	NN	O	I-DNA
and	NN	O	O
were	NN	O	O
shown	NN	O	O
by	NN	O	O
supershift	NN	O	O
to	NN	O	O
include	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
an	NN	O	O
AP1	NN	O	B-DNA
motif	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
the	NN	O	O
Jun	NN	O	B-protein
and	NN	O	I-protein
Fos	NN	O	I-protein
families	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
AP1	NN	O	B-DNA
motif	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
intron	NN	O	B-DNA
enhancer	NN	O	I-DNA
was	NN	O	O
shown	NN	O	O
to	NN	O	O
decrease	NN	O	O
enhancer-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
in	NN	O	O
both	NN	O	O
pre-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
induced	NN	O	O
with	NN	O	O
LPS	NN	O	O
and	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Distinct	NN	O	O
DNase-I	NN	O	B-DNA
hypersensitive	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
associated	NN	O	O
with	NN	O	O
TAL-1	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_line
and	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
tal-1	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
frequently	NN	O	O
activated	NN	O	O
in	NN	O	O
human	NN	O	O
T-cell	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
T-ALL	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
erythroid	NN	O	B-cell_type
,	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
mast	NN	O	I-cell_type
cell	NN	O	I-cell_type
lineages	NN	O	I-cell_type
during	NN	O	O
normal	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

To	NN	O	O
gain	NN	O	O
further	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
tal-1	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
tal-1	NN	O	B-DNA
chromatin	NN	O	O
structure	NN	O	O
in	NN	O	O
erythroid/megakaryocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
either	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
tal-1	NN	O	B-DNA
rearrangements	NN	O	I-DNA
.	NN	O	O

Tal-1	NN	O	O
transcription	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
monoallelic	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
T-cell	NN	O	O
line	NN	O	O
that	NN	O	O
expresses	NN	O	O
tal-1	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
apparent	NN	O	O
genomic	NN	O	O
alteration	NN	O	O
of	NN	O	O
the	NN	O	O
locus	NN	O	O
.	NN	O	O

Methylation	NN	O	O
studies	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
tal-15	NN	O	B-DNA
'	NN	O	I-DNA
GC-rich	NN	O	I-DNA
region	NN	O	I-DNA
behaves	NN	O	O
like	NN	O	O
a	NN	O	O
CpG	NN	O	B-DNA
island	NN	O	I-DNA
,	NN	O	O
hypomethylated	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
methylated	NN	O	O
de	NN	O	O
novo	NN	O	O
on	NN	O	O
transcriptionally	NN	O	B-DNA
inactive	NN	O	I-DNA
alleles	NN	O	I-DNA
in	NN	O	O
established	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Five	NN	O	O
major	NN	O	O
DNase-I	NN	O	B-DNA
hypersensitive	NN	O	I-DNA
sites	NN	O	I-DNA
(	NN	O	O
HS	NN	O	B-DNA
)	NN	O	O
were	NN	O	O
mapped	NN	O	O
in	NN	O	O
the	NN	O	O
tal-1	NN	O	B-DNA
locus	NN	O	O
.	NN	O	O

HS	NN	O	B-DNA
I	NN	O	I-DNA
,	NN	O	I-DNA
IV	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
V	NN	O	I-DNA
were	NN	O	O
exclusively	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
erythroid/megakaryocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
express	NN	O	O
tal-1	NN	O	B-DNA
from	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
1a	NN	O	I-DNA
and	NN	O	I-DNA
1b	NN	O	I-DNA
.	NN	O	O

HS	NN	O	B-DNA
II	NN	O	I-DNA
was	NN	O	O
weak	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
absent	NN	O	O
in	NN	O	O
Hela	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
greatly	NN	O	O
enhanced	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
region	NN	O	O
might	NN	O	O
be	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
cis-activation	NN	O	O
of	NN	O	O
tal-1	NN	O	B-DNA
promoter	NN	O	I-DNA
1b	NN	O	I-DNA
in	NN	O	O
this	NN	O	O
cell	NN	O	O
line	NN	O	O
.	NN	O	O

HS	NN	O	B-DNA
III	NN	O	I-DNA
was	NN	O	O
weak	NN	O	O
in	NN	O	O
HEL	NN	O	B-cell_line
and	NN	O	O
Jurkat	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
greatly	NN	O	O
enhanced	NN	O	O
in	NN	O	O
DU528	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
T-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
that	NN	O	O
bears	NN	O	O
a	NN	O	B-DNA
t	NN	O	I-DNA
(	NN	O	I-DNA
1	NN	O	I-DNA
;	NN	O	I-DNA
14	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
initiates	NN	O	O
tal-1	NN	O	B-DNA
transcription	NN	O	O
within	NN	O	O
exon	NN	O	B-DNA
4	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
distinct	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
are	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
different	NN	O	O
tal-1	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Erythropoietin	NN	O	B-protein
-dependent	NN	O	O
induction	NN	O	O
of	NN	O	O
hemoglobin	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
a	NN	O	O
cytokine-dependent	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
M-TAT	NN	O	B-cell_line
.	NN	O	O

M-TAT	NN	O	B-cell_line
is	NN	O	O
a	NN	O	O
cytokine-dependent	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
differentiate	NN	O	O
along	NN	O	O
the	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
cultured	NN	O	O
M-TAT	NN	O	B-cell_line
cells	NN	O	I-cell_line
long	NN	O	O
term	NN	O	O
(	NN	O	O
>	NN	O	O
1	NN	O	O
year	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
continuous	NN	O	O
presence	NN	O	O
of	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
EPO	NN	O	B-protein
)	NN	O	O
,	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
or	NN	O	O
stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
SCF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

These	NN	O	O
long	NN	O	O
term	NN	O	O
cultures	NN	O	O
are	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
M-TAT/EPO	NN	O	B-cell_line
,	NN	O	I-cell_line
M-TAT/GM-CSF	NN	O	I-cell_line
,	NN	O	I-cell_line
and	NN	O	I-cell_line
M-TAT/SCF	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Hemoglobin	NN	O	B-protein
concentration	NN	O	O
and	NN	O	O
gamma-globin	NN	O	B-protein
and	NN	O	O
erythroid	NN	O	B-RNA
delta-aminolevulinate	NN	O	I-RNA
synthase	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
were	NN	O	O
significantly	NN	O	O
higher	NN	O	O
in	NN	O	O
M-TAT/EPO	NN	O	B-cell_line
cells	NN	O	I-cell_line
than	NN	O	O
in	NN	O	O
M-TAT/GM-CSF	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

When	NN	O	O
the	NN	O	O
supplemented	NN	O	O
cytokine	NN	O	B-protein
was	NN	O	O
switched	NN	O	O
from	NN	O	O
GM-CSF	NN	O	B-protein
to	NN	O	O
EPO	NN	O	B-protein
,	NN	O	O
hemoglobin	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
M-TAT/GM-CSF	NN	O	B-cell_line
cells	NN	O	I-cell_line
increased	NN	O	O
rapidly	NN	O	O
(	NN	O	O
within	NN	O	O
5	NN	O	O
h	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
increased	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
to	NN	O	O
the	NN	O	O
M-TAT/EPO	NN	O	B-cell_line
cell	NN	O	I-cell_line
culture	NN	O	I-cell_line
decreased	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
hemoglobin	NN	O	O
,	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
EPO	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
EPO	NN	O	B-protein
signal	NN	O	O
for	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
is	NN	O	O
suppressed	NN	O	O
by	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
erythroid	NN	O	O
development	NN	O	O
of	NN	O	O
M-TAT	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
promoted	NN	O	O
by	NN	O	O
EPO	NN	O	B-protein
and	NN	O	O
suppressed	NN	O	O
by	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
EPO	NN	O	B-protein
actively	NN	O	O
influences	NN	O	O
the	NN	O	O
programming	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
required	NN	O	O
for	NN	O	O
erythroid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
E2F	NN	O	I-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cdc2	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
control	NN	O	O
of	NN	O	O
human	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

cdc2	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcripts	NN	O	I-RNA
were	NN	O	O
detected	NN	O	O
in	NN	O	O
immature	NN	O	B-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
became	NN	O	O
undetectable	NN	O	O
along	NN	O	O
with	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
resting	NN	O	I-cell_type
cells	NN	O	I-cell_type
did	NN	O	O
not	NN	O	O
express	NN	O	O
cdc2	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
but	NN	O	O
it	NN	O	O
was	NN	O	O
induced	NN	O	O
in	NN	O	O
T-lymphocytes	NN	O	B-cell_type
when	NN	O	O
the	NN	O	O
cells	NN	O	O
reentered	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
specific	NN	O	O
mitogens	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
cdc2	NN	O	B-RNA
mRNA	NN	O	I-RNA
could	NN	O	O
not	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
granulocytes	NN	O	B-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
even	NN	O	O
after	NN	O	O
the	NN	O	O
culture	NN	O	O
with	NN	O	O
the	NN	O	O
appropriate	NN	O	O
stimulants	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cdc2	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
cdc2	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
found	NN	O	O
the	NN	O	O
putative	NN	O	O
E2F	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
nucleotides	NN	O	B-DNA
-124	NN	O	I-DNA
to	NN	O	I-DNA
-117	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
at	NN	O	O
this	NN	O	O
region	NN	O	O
was	NN	O	O
detected	NN	O	O
by	NN	O	O
a	NN	O	O
gel-retardation	NN	O	O
assay	NN	O	O
and	NN	O	O
DNaseI	NN	O	B-protein
footprinting	NN	O	O
in	NN	O	O
phytohemagglutinin-stimulated	NN	O	B-cell_line
T-lymphocytes	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
was	NN	O	O
coincident	NN	O	O
with	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cdc2	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

E2F	NN	O	B-protein
binding	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
both	NN	O	O
granulocytes	NN	O	B-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Transient	NN	O	O
chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
assay	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
region	NN	O	O
containing	NN	O	O
E2F	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
had	NN	O	O
a	NN	O	O
strong	NN	O	O
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
introduction	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
at	NN	O	O
the	NN	O	O
E2F	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
significant	NN	O	O
loss	NN	O	O
of	NN	O	O
the	NN	O	O
activity	NN	O	O
.	NN	O	O

E2F-1	NN	O	B-RNA
and	NN	O	I-RNA
DP-1	NN	O	I-RNA
mRNAs	NN	O	I-RNA
were	NN	O	O
not	NN	O	O
detectable	NN	O	O
in	NN	O	O
granulocytes	NN	O	B-cell_type
,	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
resting	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
but	NN	O	O
were	NN	O	O
induced	NN	O	O
after	NN	O	O
the	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T-lymphocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
activity	NN	O	O
preceded	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
cdc2	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cdc2	NN	O	B-cell_type
mRNA	NN	O	I-cell_type
expression	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
cdc2	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
cell	NN	O	O
cycling	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
E2F	NN	O	B-protein
plays	NN	O	O
some	NN	O	O
roles	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
its	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Association	NN	O	O
of	NN	O	O
alterations	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
moieties	NN	O	I-protein
with	NN	O	O
HIV	NN	O	O
type	NN	O	O
1	NN	O	O
proviral	NN	O	O
latency	NN	O	O
in	NN	O	O
certain	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
replication	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
complex	NN	O	O
array	NN	O	O
of	NN	O	O
virally	NN	O	B-protein
encoded	NN	O	I-protein
and	NN	O	I-protein
cellular	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

A	NN	O	O
wide	NN	O	O
spectrum	NN	O	O
of	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
have	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
various	NN	O	O
cells	NN	O	O
,	NN	O	O
both	NN	O	O
in	NN	O	O
cell	NN	O	O
culture	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Molecular	NN	O	O
mechanisms	NN	O	O
leading	NN	O	O
to	NN	O	O
restricted	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
may	NN	O	O
differ	NN	O	O
between	NN	O	O
certain	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
now	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
HIV-1	NN	O	O
proviral	NN	O	O
latency	NN	O	O
in	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U1	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
which	NN	O	O
only	NN	O	O
extremely	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
are	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
baseline	NN	O	O
unstimulated	NN	O	O
state	NN	O	O
,	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
alterations	NN	O	O
in	NN	O	O
nuclear	NN	O	O
factor-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
moieties	NN	O	I-protein
demonstrated	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSAs	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
situ	NN	O	O
UV	NN	O	O
cross-linking	NN	O	O
studies	NN	O	O
.	NN	O	O

A	NN	O	O
predominance	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
moieties	NN	O	I-protein
and	NN	O	O
possibly	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	I-protein
or	NN	O	O
closely	NN	O	O
related	NN	O	O
species	NN	O	O
,	NN	O	O
rather	NN	O	O
than	NN	O	O
the	NN	O	O
p50-p56	NN	O	B-protein
heterodimer	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
that	NN	O	O
is	NN	O	O
the	NN	O	O
predominant	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
species	NN	O	O
in	NN	O	O
most	NN	O	O
T	NN	O	B-cell_type
lymphocytic	NN	O	I-cell_type
and	NN	O	I-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
pattern	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B-related	NN	O	I-protein
moieties	NN	O	I-protein
differs	NN	O	O
from	NN	O	O
the	NN	O	O
latently	NN	O	B-cell_line
infected	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
ACH-2	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
from	NN	O	O
the	NN	O	O
U937	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
parental	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
U1	NN	O	B-cell_line
cellular	NN	O	I-cell_line
clone	NN	O	I-cell_line
.	NN	O	O

As	NN	O	O
such	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
different	NN	O	O
proximal	NN	O	O
mechanisms	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
restricted	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
various	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Regulation	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
B-lymphocytes	NN	O	B-cell_type
]	NN	O	O

Since	NN	O	O
most	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
immortalized	NN	O	O
with	NN	O	O
EBV	NN	O	O
virus	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
produce	NN	O	O
interleukin-2	NN	O	B-protein
,	NN	O	O
a	NN	O	O
typical	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
cytokine	NN	O	I-protein
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
different	NN	O	O
elements	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
such	NN	O	O
clones	NN	O	O
by	NN	O	O
transfection	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
that	NN	O	O
the	NN	O	O
element	NN	O	O
TCEd	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
binds	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kB	NN	O	B-protein
,	NN	O	O
is	NN	O	O
very	NN	O	O
active	NN	O	O
in	NN	O	O
all	NN	O	O
three	NN	O	O
B	NN	O	B-cell_line
clones	NN	O	I-cell_line
tested	NN	O	O
.	NN	O	O

This	NN	O	O
element	NN	O	O
has	NN	O	O
no	NN	O	O
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
line	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
NFATd	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
binds	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NFAT-1	NN	O	B-protein
and	NN	O	O
is	NN	O	O
very	NN	O	O
active	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
only	NN	O	O
weakly	NN	O	O
active	NN	O	O
in	NN	O	O
one	NN	O	O
B	NN	O	B-cell_line
clone	NN	O	I-cell_line
and	NN	O	O
not	NN	O	O
at	NN	O	O
all	NN	O	O
in	NN	O	O
another	NN	O	O
.	NN	O	O

Different	NN	O	O
elements	NN	O	O
thus	NN	O	O
contribute	NN	O	O
to	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
different	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
p105	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
persistently	NN	O	O
infected	NN	O	O
with	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
regulating	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
persistence	NN	O	O
in	NN	O	O
human	NN	O	O
monocytes	NN	O	B-cell_type
/macrophages	NN	O	B-cell_type
are	NN	O	O
partially	NN	O	O
understood	NN	O	O
.	NN	O	O

Persistent	NN	O	O
HIV	NN	O	O
infection	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
results	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Whether	NN	O	O
virus-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
is	NN	O	O
a	NN	O	O
mechanism	NN	O	O
that	NN	O	O
favors	NN	O	O
continuous	NN	O	O
viral	NN	O	O
replication	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
remains	NN	O	O
unknown	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
delineate	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
HIV-infected	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
we	NN	O	O
have	NN	O	O
focused	NN	O	O
on	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
molecules	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
persistent	NN	O	O
HIV	NN	O	O
infection	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
not	NN	O	O
only	NN	O	O
in	NN	O	O
monocytic	NN	O	O
cells	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
HIV-infected	NN	O	O
cells	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
protein	NN	O	O
levels	NN	O	O
are	NN	O	O
decreased	NN	O	O
secondary	NN	O	O
to	NN	O	O
enhanced	NN	O	O
protein	NN	O	O
degradation	NN	O	O
.	NN	O	O

This	NN	O	O
parallels	NN	O	O
the	NN	O	O
increased	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
synthesis	NN	O	O
secondary	NN	O	O
to	NN	O	O
increased	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
increased	NN	O	O
RNA	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
its	NN	O	O
promoter-enhancer	NN	O	O
.	NN	O	O

Another	NN	O	O
protein	NN	O	O
with	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
function	NN	O	O
,	NN	O	O
p105	NN	O	B-protein
,	NN	O	O
is	NN	O	O
also	NN	O	O
modified	NN	O	O
in	NN	O	O
HIV-infected	NN	O	O
cells	NN	O	O
:	NN	O	O
p105	NN	O	B-protein
and	NN	O	O
p50	NN	O	B-protein
steady-state	NN	O	O
protein	NN	O	O
levels	NN	O	O
are	NN	O	O
increased	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
increased	NN	O	O
synthesis	NN	O	O
and	NN	O	O
proteolytic	NN	O	O
processing	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
.	NN	O	O

Transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
is	NN	O	O
also	NN	O	O
increased	NN	O	O
in	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
also	NN	O	O
mediated	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
through	NN	O	O
a	NN	O	O
specific	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
triple	NN	O	O
autoregulatory	NN	O	O
loop	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
involving	NN	O	O
HIV	NN	O	O
,	NN	O	O
p105	NN	O	B-protein
and	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
and	NN	O	O
MAD3	NN	O	B-protein
,	NN	O	O
with	NN	O	O
the	NN	O	O
end	NN	O	O
result	NN	O	O
of	NN	O	O
persistent	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
activation	NN	O	O
and	NN	O	O
viral	NN	O	O
persistence	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
persistent	NN	O	O
HIV	NN	O	O
infection	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
provides	NN	O	O
a	NN	O	O
useful	NN	O	O
model	NN	O	O
with	NN	O	O
which	NN	O	O
to	NN	O	O
study	NN	O	O
concomitant	NN	O	O
modifications	NN	O	O
of	NN	O	O
different	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Glucocorticoid-induced	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
repressive	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
which	NN	O	O
may	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
therapeutic	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
various	NN	O	O
diseases	NN	O	O
including	NN	O	O
leukemia	NN	O	O
,	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
events	NN	O	O
leading	NN	O	O
from	NN	O	O
the	NN	O	O
activated	NN	O	O
receptor	NN	O	O
to	NN	O	O
cell	NN	O	O
lysis	NN	O	O
are	NN	O	O
not	NN	O	O
understood	NN	O	O
.	NN	O	O

A	NN	O	O
prevailing	NN	O	O
hypothesis	NN	O	O
postulates	NN	O	O
induction	NN	O	O
of	NN	O	O
so-called	NN	O	O
'	NN	O	O
lysis	NN	O	B-DNA
genes	NN	O	I-DNA
'	NN	O	O
by	NN	O	O
the	NN	O	O
activated	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
an	NN	O	O
activation-deficient	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
mutant	NN	O	I-protein
is	NN	O	O
as	NN	O	O
effective	NN	O	O
as	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
repression	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
production	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
c-myc	NN	O	B-DNA
expression	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
can	NN	O	O
also	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
through	NN	O	O
the	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
whose	NN	O	O
repressive	NN	O	O
functions	NN	O	O
but	NN	O	O
not	NN	O	O
target	NN	O	O
site	NN	O	O
specificity	NN	O	O
,	NN	O	O
are	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
primary	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
in	NN	O	O
glucocorticoid-mediated	NN	O	O
apoptosis	NN	O	O
correlates	NN	O	O
with	NN	O	O
transcriptional	NN	O	O
repression	NN	O	O
rather	NN	O	O
than	NN	O	O
activation	NN	O	O
and	NN	O	O
could	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
interference	NN	O	O
with	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
required	NN	O	O
for	NN	O	O
cell	NN	O	O
survival	NN	O	O
.	NN	O	O

-DOCSTART-	O

HIV	NN	O	O
type	NN	O	O
1	NN	O	O
protease	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
within	NN	O	O
T	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
the	NN	O	O
rel	NN	O	B-protein
oncogene	NN	O	I-protein
family	NN	O	I-protein
capable	NN	O	O
of	NN	O	O
enhancing	NN	O	O
transcription	NN	O	O
of	NN	O	O
several	NN	O	B-DNA
cellular	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
viral	NN	O	B-DNA
genes	NN	O	I-DNA
transcribed	NN	O	O
from	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
that	NN	O	O
HIV-1	NN	O	B-protein
protease	NN	O	I-protein
may	NN	O	O
cleave	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
precursor	NN	O	O
to	NN	O	O
its	NN	O	O
active	NN	O	O
form	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
HIV	NN	O	O
protease	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
precursor	NN	O	O
activation	NN	O	O
were	NN	O	O
examined	NN	O	O
in	NN	O	O
Jurkat	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
introducing	NN	O	O
a	NN	O	O
protease	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
into	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

Increased	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
was	NN	O	O
observed	NN	O	O
and	NN	O	O
this	NN	O	O
increased	NN	O	O
activity	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
a	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
protease	NN	O	I-protein
.	NN	O	O

Viral	NN	O	O
transcription	NN	O	O
,	NN	O	O
as	NN	O	O
measured	NN	O	O
using	NN	O	O
LTR	NN	O	B-DNA
-CAT	NN	O	B-protein
assays	NN	O	O
,	NN	O	O
was	NN	O	O
only	NN	O	O
slightly	NN	O	O
enhanced	NN	O	O
in	NN	O	O
the	NN	O	O
HIV-protease	NN	O	B-cell_type
expressing	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	O
were	NN	O	O
not	NN	O	O
affected	NN	O	O
.	NN	O	O

The	NN	O	O
limited	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
HIV	NN	O	B-protein
protease	NN	O	I-protein
appears	NN	O	O
unlikely	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
significant	NN	O	O
effect	NN	O	O
on	NN	O	O
virus	NN	O	O
expression	NN	O	O
or	NN	O	O
T	NN	O	O
cell	NN	O	O
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
replication	NN	O	O
by	NN	O	O
a	NN	O	O
Tat-activated	NN	O	B-DNA
,	NN	O	I-DNA
transduced	NN	O	I-DNA
interferon	NN	O	I-DNA
gene	NN	O	I-DNA
:	NN	O	O
targeted	NN	O	O
expression	NN	O	O
to	NN	O	O
human	NN	O	B-cell_type
immunodeficiency	NN	O	I-cell_type
virus	NN	O	I-cell_type
type	NN	O	I-cell_type
1-infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
feasibility	NN	O	O
of	NN	O	O
using	NN	O	O
interferon	NN	O	B-protein
(	NN	O	O
IFN	NN	O	B-protein
)	NN	O	O
gene	NN	O	O
transfer	NN	O	O
as	NN	O	O
a	NN	O	O
novel	NN	O	O
approach	NN	O	O
to	NN	O	O
anti-human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
therapy	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
.	NN	O	O

To	NN	O	O
limit	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
transduced	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	I-DNA
LTR	NN	O	I-DNA
)	NN	O	I-DNA
-IFNA2	NN	O	I-DNA
(	NN	O	O
the	NN	O	O
new	NN	O	O
approved	NN	O	O
nomenclature	NN	O	O
for	NN	O	O
IFN	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
used	NN	O	O
throughout	NN	O	O
this	NN	O	O
article	NN	O	O
)	NN	O	O
hybrid	NN	O	B-DNA
gene	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
HIV-1-infected	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
was	NN	O	O
modified	NN	O	O
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
significantly	NN	O	O
inhibited	NN	O	O
Tat	NN	O	B-protein
-mediated	NN	O	O
transactivation	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
a	NN	O	O
monocyte	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
U937	NN	O	B-cell_line
.	NN	O	O

Replacement	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
by	NN	O	O
a	NN	O	O
DNA	NN	O	B-DNA
fragment	NN	O	I-DNA
derived	NN	O	O
from	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
IFN-stimulated	NN	O	B-DNA
gene	NN	O	I-DNA
15	NN	O	I-DNA
(	NN	O	O
ISG15	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
containing	NN	O	O
the	NN	O	O
IFN-stimulated	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
partially	NN	O	O
restored	NN	O	O
Tat	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
LTR	NN	O	B-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Insertion	NN	O	O
of	NN	O	O
this	NN	O	O
chimeric	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
ISG15	NN	O	B-DNA
LTR	NN	O	I-DNA
)	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IFNA2	NN	O	I-DNA
gene	NN	O	I-DNA
directed	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
IFN	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
Tat-expressing	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
this	NN	O	O
promoter	NN	O	O
was	NN	O	O
not	NN	O	O
responsive	NN	O	O
to	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
-mediated	NN	O	O
activation	NN	O	O
.	NN	O	O

ISG15-LTR-IFN	NN	O	B-DNA
hybrid	NN	O	I-DNA
gene	NN	O	I-DNA
inserted	NN	O	O
into	NN	O	O
the	NN	O	O
retrovirus	NN	O	B-DNA
vector	NN	O	I-DNA
was	NN	O	O
transduced	NN	O	O
into	NN	O	O
Jurkat	NN	O	B-cell_line
and	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Selected	NN	O	O
transfected	NN	O	B-cell_line
clones	NN	O	I-cell_line
produced	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
IFN	NN	O	B-protein
A	NN	O	I-protein
(	NN	O	O
IFNA	NN	O	B-protein
)	NN	O	O
constitutively	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
abilities	NN	O	O
to	NN	O	O
express	NN	O	O
interleukin-2	NN	O	B-protein
and	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phytohemagglutinin	NN	O	B-protein
and	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
were	NN	O	O
retained	NN	O	O
.	NN	O	O

Enhancement	NN	O	O
of	NN	O	O
IFNA	NN	O	B-protein
synthesis	NN	O	O
observed	NN	O	O
upon	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
resulted	NN	O	O
in	NN	O	O
significant	NN	O	O
inhibition	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
for	NN	O	O
a	NN	O	O
period	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
30	NN	O	O
days	NN	O	O
.	NN	O	O

Virus	NN	O	O
isolated	NN	O	O
from	NN	O	O
IFNA-producing	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
able	NN	O	O
to	NN	O	O
replicate	NN	O	O
in	NN	O	O
the	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
did	NN	O	O
not	NN	O	O
replicate	NN	O	O
efficiently	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
transduced	NN	O	O
with	NN	O	O
the	NN	O	O
IFNA	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
targeting	NN	O	O
IFN	NN	O	B-protein
synthesis	NN	O	O
to	NN	O	O
HIV-1-infected	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
an	NN	O	O
attainable	NN	O	O
goal	NN	O	O
and	NN	O	O
that	NN	O	O
autocrine	NN	O	O
IFN	NN	O	B-protein
synthesis	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
long-lasting	NN	O	O
and	NN	O	O
permanent	NN	O	O
suppression	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

-DOCSTART-	O

Chromosomal	NN	O	O
localization	NN	O	O
of	NN	O	O
two	NN	O	O
KOX	NN	O	B-DNA
zinc	NN	O	I-DNA
finger	NN	O	I-DNA
genes	NN	O	I-DNA
on	NN	O	O
chromosome	NN	O	B-DNA
bands	NN	O	I-DNA
7q21-q22	NN	O	B-DNA
.	NN	O	O

Human	NN	O	B-DNA
cDNAs	NN	O	I-DNA
encoding	NN	O	O
Kruppel-type	NN	O	B-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
domains	NN	O	I-protein
,	NN	O	O
designated	NN	O	O
KOX	NN	O	B-DNA
1-32	NN	O	I-DNA
,	NN	O	O
have	NN	O	O
been	NN	O	O
cloned	NN	O	O
from	NN	O	O
human	NN	O	B-DNA
T	NN	O	I-DNA
lymphocyte	NN	O	I-DNA
cell	NN	O	I-DNA
line	NN	O	I-DNA
libraries	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
regional	NN	O	O
localizations	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
of	NN	O	O
KOX	NN	O	B-DNA
18	NN	O	I-DNA
and	NN	O	O
KOX	NN	O	B-DNA
25	NN	O	I-DNA
on	NN	O	O
chromosome	NN	O	B-DNA
bands	NN	O	I-DNA
7q21q22	NN	O	I-DNA
.	NN	O	O

Pulse-field	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
(	NN	O	O
PFGE	NN	O	O
)	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
these	NN	O	O
genes	NN	O	O
are	NN	O	O
physically	NN	O	O
located	NN	O	O
within	NN	O	O
a	NN	O	O
DNA	NN	O	B-DNA
fragment	NN	O	I-DNA
of	NN	O	O
250	NN	O	B-DNA
kb	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
genes	NN	O	O
KOX	NN	O	B-DNA
4	NN	O	I-DNA
and	NN	O	O
KOX	NN	O	B-DNA
9	NN	O	I-DNA
,	NN	O	O
mapped	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
8q24	NN	O	I-DNA
,	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
located	NN	O	O
within	NN	O	O
a	NN	O	O
DNA	NN	O	B-DNA
fragment	NN	O	I-DNA
of	NN	O	O
450	NN	O	O
kb	NN	O	O
.	NN	O	O

From	NN	O	O
the	NN	O	O
present	NN	O	O
and	NN	O	O
previous	NN	O	O
data	NN	O	O
,	NN	O	O
eighteen	NN	O	O
different	NN	O	O
KOX	NN	O	B-DNA
genes	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
located	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
by	NN	O	O
two	NN	O	O
within	NN	O	O
nine	NN	O	O
DNA	NN	O	B-DNA
fragments	NN	O	I-DNA
of	NN	O	O
200	NN	O	O
to	NN	O	O
580	NN	O	O
kb	NN	O	O
.	NN	O	O

-DOCSTART-	O

Influence	NN	O	O
of	NN	O	O
age	NN	O	O
on	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
by	NN	O	O
influenza	NN	O	B-cell_type
virus-exposed	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
study	NN	O	O
investigated	NN	O	O
age-related	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
after	NN	O	O
in	NN	O	O
vitro	NN	O	O
exposure	NN	O	O
to	NN	O	O
influenza	NN	O	O
A	NN	O	O
virus	NN	O	O
.	NN	O	O

Mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
from	NN	O	O
young	NN	O	O
or	NN	O	O
elderly	NN	O	O
persons	NN	O	O
were	NN	O	O
sham-exposed	NN	O	O
or	NN	O	O
exposed	NN	O	O
to	NN	O	O
influenza	NN	O	O
virus	NN	O	O
for	NN	O	O
1	NN	O	O
,	NN	O	O
24	NN	O	O
,	NN	O	O
and	NN	O	O
72	NN	O	O
h	NN	O	O
.	NN	O	O

Immunofluorescent	NN	O	O
staining	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometric	NN	O	O
analysis	NN	O	O
were	NN	O	O
then	NN	O	O
used	NN	O	O
to	NN	O	O
detect	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
producing	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
regulating	NN	O	I-protein
proteins	NN	O	I-protein
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
.	NN	O	O

Fewer	NN	O	O
virus-exposed	NN	O	O
cells	NN	O	O
from	NN	O	O
elderly	NN	O	O
donors	NN	O	O
stained	NN	O	O
for	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
at	NN	O	O
each	NN	O	O
data	NN	O	O
point	NN	O	O
compared	NN	O	O
with	NN	O	O
cells	NN	O	O
from	NN	O	O
young	NN	O	O
donors	NN	O	O
.	NN	O	O

Flow	NN	O	O
cytometric	NN	O	O
analysis	NN	O	O
also	NN	O	O
showed	NN	O	O
that	NN	O	O
at	NN	O	O
72	NN	O	O
h	NN	O	O
of	NN	O	O
virus	NN	O	O
exposure	NN	O	O
fewer	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
elderly	NN	O	O
produced	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
link	NN	O	O
between	NN	O	O
the	NN	O	O
magnitude	NN	O	O
of	NN	O	O
the	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
responses	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
failure	NN	O	O
of	NN	O	O
virus-exposed	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
produce	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
could	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
illness	NN	O	O
due	NN	O	O
to	NN	O	O
influenza	NN	O	O
virus	NN	O	O
in	NN	O	O
the	NN	O	O
elderly	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
HIV	NN	O	O
by	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
correlates	NN	O	O
with	NN	O	O
cellular	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

OBJECTIVES	NN	O	O
:	NN	O	O
To	NN	O	O
analyze	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
retinoic	NN	O	O
acids	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
and	NN	O	O
correlate	NN	O	O
this	NN	O	O
effect	NN	O	O
with	NN	O	O
expression	NN	O	O
levels	NN	O	O
of	NN	O	O
RA	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
RARs	NN	O	B-protein
)	NN	O	O
in	NN	O	O
T-lymphoid	NN	O	B-cell_line
and	NN	O	I-cell_line
monocytoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

DESIGN	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
The	NN	O	O
effect	NN	O	O
of	NN	O	O
all-trans	NN	O	O
and	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
in	NN	O	O
T-lymphoid	NN	O	B-cell_line
(	NN	O	O
H9	NN	O	B-cell_line
,	NN	O	I-cell_line
CEM	NN	O	I-cell_line
)	NN	O	O
and	NN	O	O
monocytoid	NN	O	B-cell_line
(	NN	O	I-cell_line
U937	NN	O	I-cell_line
,	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
was	NN	O	O
measured	NN	O	O
during	NN	O	O
acute	NN	O	O
and	NN	O	O
chronic	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
levels	NN	O	O
of	NN	O	O
human	NN	O	B-protein
RAR	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
hRAR	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	I-protein
receptor	NN	O	I-protein
for	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
human	NN	O	B-protein
retinoid-X	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
hRXR	NN	O	B-protein
alpha	NN	O	I-protein
receptor	NN	O	I-protein
for	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
)	NN	O	O
were	NN	O	O
determined	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Both	NN	O	O
all-trans	NN	O	O
and	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
inhibited	NN	O	O
virus	NN	O	O
replication	NN	O	O
in	NN	O	O
HIV-1	NN	O	B-cell_line
IIIB-infected	NN	O	I-cell_line
monocytoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
and	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
co-stimulatory	NN	O	O
agent	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
retinoids	NN	O	O
had	NN	O	O
weak	NN	O	O
or	NN	O	O
no	NN	O	O
stimulatory	NN	O	O
effects	NN	O	O
on	NN	O	O
HIV	NN	O	O
production	NN	O	O
by	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

HIV	NN	O	O
production	NN	O	O
by	NN	O	O
PMA-stimulated	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
these	NN	O	O
retinoids	NN	O	O
.	NN	O	O

The	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
was	NN	O	O
generally	NN	O	O
more	NN	O	O
effective	NN	O	O
than	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
in	NN	O	O
inhibiting	NN	O	O
HIV	NN	O	O
production	NN	O	O
and	NN	O	O
in	NN	O	O
combination	NN	O	O
generally	NN	O	O
more	NN	O	O
effective	NN	O	O
than	NN	O	O
the	NN	O	O
single	NN	O	O
agents	NN	O	O
alone	NN	O	O
.	NN	O	O

Human	NN	O	B-protein
RAR	NN	O	I-protein
alpha	NN	O	I-protein
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
H9	NN	O	B-cell_line
,	NN	O	O
U937	NN	O	B-cell_line
and	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
almost	NN	O	O
undetectable	NN	O	O
in	NN	O	O
CEM	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Human	NN	O	B-protein
RXR	NN	O	I-protein
alpha	NN	O	I-protein
was	NN	O	O
significantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
and	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
weakly	NN	O	O
expressed	NN	O	O
in	NN	O	O
H9	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
not	NN	O	O
detectable	NN	O	O
in	NN	O	O
CEM	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

After	NN	O	O
stimulation	NN	O	O
by	NN	O	O
PMA	NN	O	O
,	NN	O	O
RXR	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
increased	NN	O	O
in	NN	O	O
H9	NN	O	B-cell_line
and	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
CEM	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Human	NN	O	O
RAR	NN	O	O
alpha	NN	O	O
expression	NN	O	O
was	NN	O	O
unchanged	NN	O	O
in	NN	O	O
H9	NN	O	B-cell_line
and	NN	O	O
CEM	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
elevated	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
after	NN	O	O
PMA	NN	O	O
stimulation	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
The	NN	O	O
effect	NN	O	O
of	NN	O	O
RA	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
was	NN	O	O
cell-type-dependent	NN	O	O
and	NN	O	O
partially	NN	O	O
correlated	NN	O	O
with	NN	O	O
cellular	NN	O	O
expression	NN	O	O
of	NN	O	O
RARs	NN	O	B-protein
.	NN	O	O

Endogenous	NN	O	O
or	NN	O	O
exogenously	NN	O	O
administered	NN	O	O
RA	NN	O	O
may	NN	O	O
have	NN	O	O
a	NN	O	O
significant	NN	O	O
role	NN	O	O
in	NN	O	O
HIV	NN	O	O
regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functions	NN	O	O
of	NN	O	O
glutathione	NN	O	O
and	NN	O	O
glutathione	NN	O	O
disulfide	NN	O	O
in	NN	O	O
immunology	NN	O	O
and	NN	O	O
immunopathology	NN	O	O
.	NN	O	O

Even	NN	O	O
a	NN	O	O
moderate	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
cysteine	NN	O	O
supply	NN	O	O
elevates	NN	O	O
the	NN	O	O
intracellular	NN	O	O
glutathione	NN	O	O
(	NN	O	O
GSH	NN	O	O
)	NN	O	O
and	NN	O	O
glutathione	NN	O	O
disulfide	NN	O	O
(	NN	O	O
GSSG	NN	O	O
)	NN	O	O
levels	NN	O	O
and	NN	O	O
potentiates	NN	O	O
immunological	NN	O	O
functions	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

At	NN	O	O
low	NN	O	O
GSSG	NN	O	O
levels	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
not	NN	O	O
optimally	NN	O	O
activate	NN	O	O
the	NN	O	O
immunologically	NN	O	O
important	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
high	NN	O	O
GSSG	NN	O	O
levels	NN	O	O
inhibit	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
GSSG	NN	O	O
are	NN	O	O
antagonized	NN	O	O
by	NN	O	O
reduced	NN	O	O
thioredoxin	NN	O	B-protein
(	NN	O	O
TRX	NN	O	B-protein
)	NN	O	O
.	NN	O	O

As	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
activities	NN	O	O
p56lck	NN	O	B-protein
and	NN	O	O
p59fyn	NN	O	B-protein
are	NN	O	O
activated	NN	O	O
in	NN	O	O
intact	NN	O	O
cells	NN	O	O
by	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
,	NN	O	O
they	NN	O	O
are	NN	O	O
likely	NN	O	O
targets	NN	O	O
for	NN	O	O
GSSG	NN	O	O
action	NN	O	O
.	NN	O	O

These	NN	O	O
redox-regulated	NN	O	B-protein
enzymes	NN	O	I-protein
trigger	NN	O	O
signal	NN	O	O
cascades	NN	O	O
for	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
transduce	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
from	NN	O	O
CD4	NN	O	B-protein
and	NN	O	O
CD8	NN	O	B-protein
molecules	NN	O	I-protein
,	NN	O	O
and	NN	O	O
from	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
beta-chain	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effector	NN	O	O
phase	NN	O	O
of	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
responses	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
-dependent	NN	O	O
functions	NN	O	O
are	NN	O	O
inhibited	NN	O	O
even	NN	O	O
by	NN	O	O
a	NN	O	O
partial	NN	O	O
depletion	NN	O	O
of	NN	O	O
the	NN	O	O
intracellular	NN	O	O
GSH	NN	O	O
pool	NN	O	O
.	NN	O	O

As	NN	O	O
signal	NN	O	O
transduction	NN	O	O
is	NN	O	O
facilitated	NN	O	O
by	NN	O	O
prooxidant	NN	O	O
conditions	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
well-known	NN	O	O
immunological	NN	O	O
consequences	NN	O	O
of	NN	O	O
GSH	NN	O	O
depletion	NN	O	O
ultimately	NN	O	O
may	NN	O	O
be	NN	O	O
results	NN	O	O
of	NN	O	O
the	NN	O	O
accompanying	NN	O	O
GSSG	NN	O	O
deficiency	NN	O	O
.	NN	O	O

As	NN	O	O
HIV-infected	NN	O	O
patients	NN	O	O
and	NN	O	O
SIV-infected	NN	O	O
rhesus	NN	O	O
macaques	NN	O	O
have	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
average	NN	O	O
,	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
plasma	NN	O	O
cyst	NN	O	O
(	NN	O	O
e	NN	O	O
)	NN	O	O
ine	NN	O	O
and	NN	O	O
intracellular	NN	O	O
GSH	NN	O	O
levels	NN	O	O
,	NN	O	O
we	NN	O	O
also	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
AIDS	NN	O	O
may	NN	O	O
be	NN	O	O
the	NN	O	O
consequence	NN	O	O
of	NN	O	O
a	NN	O	O
GSSG	NN	O	O
deficiency	NN	O	O
as	NN	O	O
well	NN	O	O
.	NN	O	O

-DOCSTART-	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
renal	NN	O	O
cell	NN	O	O
carcinoma	NN	O	O
patients	NN	O	O
exhibit	NN	O	O
an	NN	O	O
abnormal	NN	O	O
pattern	NN	O	O
of	NN	O	O
kappa	NN	O	O
B-specific	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
:	NN	O	O
a	NN	O	O
preliminary	NN	O	O
report	NN	O	O
.	NN	O	O

Recent	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
poor	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
T-cell	NN	O	O
response	NN	O	O
to	NN	O	O
human	NN	O	O
renal	NN	O	O
cell	NN	O	O
carcinoma	NN	O	O
(	NN	O	O
RCC	NN	O	O
)	NN	O	O
may	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
alterations	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
RCC	NN	O	O
patients	NN	O	O
have	NN	O	O
two	NN	O	O
alterations	NN	O	O
in	NN	O	O
kappa	NN	O	O
B	NN	O	O
motif-specific	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
alteration	NN	O	O
involves	NN	O	O
the	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
substantial	NN	O	O
kappa	NN	O	O
B-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

The	NN	O	O
magnitude	NN	O	O
of	NN	O	O
kappa	NN	O	O
B	NN	O	O
activity	NN	O	O
in	NN	O	O
unstimulated	NN	O	B-cell_type
patient	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
observed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
normal	NN	O	O
individuals	NN	O	O
that	NN	O	O
had	NN	O	O
been	NN	O	O
activated	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
Western	NN	O	O
blotting	NN	O	O
experiments	NN	O	O
using	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	O
kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
the	NN	O	O
kappa	NN	O	O
B-binding	NN	O	O
activity	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
RCC	NN	O	O
patients	NN	O	O
is	NN	O	O
composed	NN	O	O
mostly	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
(	NN	O	I-protein
p50	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
.	NN	O	O

The	NN	O	O
second	NN	O	O
abnormality	NN	O	O
in	NN	O	O
kappa	NN	O	O
B-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
these	NN	O	O
patients	NN	O	O
is	NN	O	O
that	NN	O	O
RelA	NN	O	B-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
Rel	NN	O	B-protein
homology	NN	O	I-protein
family	NN	O	I-protein
which	NN	O	O
is	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
was	NN	O	O
not	NN	O	O
induced	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
following	NN	O	O
activation	NN	O	O
.	NN	O	O

Western	NN	O	O
blotting	NN	O	O
analysis	NN	O	O
did	NN	O	O
not	NN	O	O
detect	NN	O	O
any	NN	O	O
RelA	NN	O	B-protein
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
either	NN	O	O
before	NN	O	O
or	NN	O	O
after	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
altered	NN	O	O
kappa	NN	O	O
B-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
RCC	NN	O	O
patients	NN	O	O
may	NN	O	O
impair	NN	O	O
their	NN	O	O
capacity	NN	O	O
to	NN	O	O
respond	NN	O	O
normally	NN	O	O
to	NN	O	O
various	NN	O	O
stimuli	NN	O	O
.	NN	O	O

-DOCSTART-	O

Hemoglobin	NN	O	B-protein
switching	NN	O	O
in	NN	O	O
humans	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
SP1	NN	O	B-protein
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Understanding	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
developmental	NN	O	O
regulation	NN	O	O
of	NN	O	O
hemoglobin	NN	O	B-protein
switching	NN	O	O
has	NN	O	O
scientific	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
clinical	NN	O	O
relevance	NN	O	O
because	NN	O	O
of	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
fetal	NN	O	O
hemoglobin	NN	O	B-protein
(	NN	O	O
HbF	NN	O	B-protein
)	NN	O	O
production	NN	O	O
in	NN	O	O
adulthood	NN	O	O
on	NN	O	O
the	NN	O	O
clinical	NN	O	O
manifestation	NN	O	O
of	NN	O	O
thalassemia	NN	O	O
and	NN	O	O
sickle	NN	O	O
cell	NN	O	O
anemia	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
normal	NN	O	O
developmental	NN	O	O
patterns	NN	O	O
of	NN	O	O
globin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
are	NN	O	O
recapitulated	NN	O	O
in	NN	O	O
an	NN	O	O
experimental	NN	O	O
system	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_line
cultures	NN	O	I-cell_line
that	NN	O	O
support	NN	O	O
differentiation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
further	NN	O	O
found	NN	O	O
that	NN	O	O
high	NN	O	O
activities	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
activators	NN	O	I-protein
,	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
SP1	NN	O	B-protein
,	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
normal	NN	O	O
adult	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

MATERIALS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
In	NN	O	O
the	NN	O	O
present	NN	O	O
work	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
,	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
SP1	NN	O	B-protein
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
cultured	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
derived	NN	O	O
from	NN	O	O
cord	NN	O	O
blood	NN	O	O
and	NN	O	O
from	NN	O	O
fetal	NN	O	O
livers	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
from	NN	O	O
beta	NN	O	O
zero-thalassemia	NN	O	O
patients	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
The	NN	O	O
results	NN	O	O
showed	NN	O	O
high	NN	O	O
GATA-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
very	NN	O	O
low	NN	O	O
SP1	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
the	NN	O	O
fetal	NN	O	B-cell_line
liver	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
pattern	NN	O	O
was	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
cultures	NN	O	O
derived	NN	O	O
from	NN	O	O
normal	NN	O	O
adult	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
both	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
SP1	NN	O	B-protein
activities	NN	O	O
were	NN	O	O
high	NN	O	O
.	NN	O	O

Cord	NN	O	O
blood	NN	O	O
cultures	NN	O	O
showed	NN	O	O
an	NN	O	O
additive	NN	O	O
combination	NN	O	O
of	NN	O	O
``	NN	O	O
adult	NN	O	O
''	NN	O	O
and	NN	O	O
``	NN	O	O
fetal	NN	O	O
''	NN	O	O
patterns	NN	O	O
.	NN	O	O

The	NN	O	O
progenitors	NN	O	O
derived	NN	O	O
from	NN	O	O
a	NN	O	O
beta	NN	O	O
zero-thalassemia	NN	O	O
patient	NN	O	O
with	NN	O	O
high	NN	O	O
HbF	NN	O	B-protein
production	NN	O	O
showed	NN	O	O
``	NN	O	O
fetal	NN	O	O
''	NN	O	O
pattern	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
in	NN	O	O
cultures	NN	O	O
of	NN	O	O
2	NN	O	O
beta	NN	O	O
zero-thalassemia	NN	O	O
patients	NN	O	O
without	NN	O	O
high	NN	O	O
HbF	NN	O	B-protein
,	NN	O	O
``	NN	O	O
adult	NN	O	O
''	NN	O	O
pattern	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
In	NN	O	O
the	NN	O	O
present	NN	O	O
work	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
fetal	NN	O	I-cell_type
and	NN	O	I-cell_type
adult	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
are	NN	O	O
distinct	NN	O	O
in	NN	O	O
their	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
commitment	NN	O	O
to	NN	O	O
fetal	NN	O	O
or	NN	O	O
adult	NN	O	O
program	NN	O	O
occurs	NN	O	O
at	NN	O	O
a	NN	O	O
very	NN	O	O
early	NN	O	O
differentiation	NN	O	O
stage	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
under	NN	O	O
anemic	NN	O	O
stress	NN	O	O
,	NN	O	O
recruitment	NN	O	O
of	NN	O	O
fetal	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
may	NN	O	O
occur	NN	O	O
in	NN	O	O
adulthood	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription-independent	NN	O	O
turnover	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
during	NN	O	O
monocyte	NN	O	O
adherence	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
a	NN	O	O
translational	NN	O	O
component	NN	O	O
regulating	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
alpha/MAD-3	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha/MAD-3	NN	O	I-protein
as	NN	O	O
an	NN	O	O
immediate-early	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
signals	NN	O	O
,	NN	O	O
including	NN	O	O
adhesion	NN	O	O
,	NN	O	O
lipopolysaccharide	NN	O	O
,	NN	O	O
and	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
.	NN	O	O

Within	NN	O	O
5	NN	O	O
min	NN	O	O
of	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
,	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
markedly	NN	O	O
diminished	NN	O	O
but	NN	O	O
is	NN	O	O
rapidly	NN	O	O
replaced	NN	O	O
in	NN	O	O
a	NN	O	O
cycloheximide-sensitive	NN	O	O
manner	NN	O	O
within	NN	O	O
20	NN	O	O
min	NN	O	O
.	NN	O	O

Accompanying	NN	O	O
the	NN	O	O
rapid	NN	O	O
turnover	NN	O	O
of	NN	O	O
the	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
simultaneous	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B-related	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
to	NN	O	O
nuclei	NN	O	O
of	NN	O	O
adhered	NN	O	O
monocytes	NN	O	O
.	NN	O	O

The	NN	O	O
demonstration	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
can	NN	O	O
regulate	NN	O	O
I	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
alpha/MAD-3	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
rapid	NN	O	O
increase	NN	O	O
in	NN	O	O
steady-state	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
alpha/MAD-3	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
we	NN	O	O
observed	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
of	NN	O	O
monocyte	NN	O	O
adherence	NN	O	O
would	NN	O	O
result	NN	O	O
from	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
transcriptional	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
I	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
alpha/MAD-3	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Nuclear	NN	O	O
run-on	NN	O	O
analyses	NN	O	O
indicated	NN	O	O
that	NN	O	O
,	NN	O	O
instead	NN	O	O
,	NN	O	O
while	NN	O	O
several	NN	O	O
immediate-early	NN	O	B-DNA
cytokine	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
1	NN	O	I-DNA
beta	NN	O	I-DNA
(	NN	O	I-DNA
IL-1	NN	O	I-DNA
beta	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
were	NN	O	O
transcriptionally	NN	O	O
activated	NN	O	O
during	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
,	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
I	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
alpha/MAD-3	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
remained	NN	O	O
constant	NN	O	O
.	NN	O	O

The	NN	O	O
adherence-dependent	NN	O	O
increase	NN	O	O
in	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
alpha/MAD-3	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
was	NN	O	O
also	NN	O	O
not	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
stabilization	NN	O	O
events	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
while	NN	O	O
increases	NN	O	O
in	NN	O	O
both	NN	O	O
IL-1	NN	O	O
beta	NN	O	O
and	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
alpha/MAD-3	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
nuclei	NN	O	O
of	NN	O	O
adherent	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
cytoplasmic	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-1	NN	O	B-RNA
beta	NN	O	I-RNA
mRNA	NN	O	I-RNA
increased	NN	O	O
during	NN	O	O
adherence	NN	O	O
whereas	NN	O	O
those	NN	O	O
of	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
alpha/MAD-3	NN	O	I-RNA
mRNA	NN	O	I-RNA
did	NN	O	O
not	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
two	NN	O	O
interactive	NN	O	O
mechanisms	NN	O	O
regulate	NN	O	O
monocytic	NN	O	B-RNA
I	NN	O	I-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
alpha/MAD-3	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
adherent	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
regulate	NN	O	O
nuclear	NN	O	O
processing	NN	O	O
(	NN	O	O
or	NN	O	O
decay	NN	O	O
)	NN	O	O
of	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
alpha/MAD-3	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
thereby	NN	O	O
increasing	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
without	NN	O	O
stimulating	NN	O	O
I	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
alpha/MAD-3	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
since	NN	O	O
inhibition	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
leads	NN	O	O
to	NN	O	O
accumulation	NN	O	O
of	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
alpha/MAD-3	NN	O	I-RNA
mRNA	NN	O	I-RNA
without	NN	O	O
stimulating	NN	O	O
I	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
alpha/MAD-3	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
,	NN	O	O
we	NN	O	O
suggest	NN	O	O
that	NN	O	O
low	NN	O	O
cytoplasmic	NN	O	O
levels	NN	O	O
of	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
alpha/MAD-3	NN	O	I-RNA
mRNA	NN	O	I-RNA
are	NN	O	O
maintained	NN	O	O
by	NN	O	O
a	NN	O	O
translation-dependent	NN	O	O
degradation	NN	O	O
mechanism	NN	O	O
.	NN	O	O

-DOCSTART-	O

Distinct	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
chaperone	NN	O	B-protein
hsp90	NN	O	I-protein
in	NN	O	O
modulating	NN	O	O
dioxin	NN	O	O
receptor	NN	O	O
function	NN	O	O
via	NN	O	O
the	NN	O	O
basic	NN	O	O
helix-loop-helix	NN	O	B-protein
and	NN	O	O
PAS	NN	O	B-protein
domains	NN	O	I-protein
.	NN	O	O

The	NN	O	O
intracellular	NN	O	B-protein
dioxin	NN	O	I-protein
receptor	NN	O	I-protein
mediates	NN	O	O
signal	NN	O	O
transduction	NN	O	O
by	NN	O	O
dioxin	NN	O	O
and	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
ligand-activated	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

It	NN	O	O
contains	NN	O	O
a	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
(	NN	O	I-protein
bHLH	NN	O	I-protein
)	NN	O	I-protein
motif	NN	O	I-protein
contiguous	NN	O	O
with	NN	O	O
a	NN	O	O
Per-Arnt-Sim	NN	O	B-protein
(	NN	O	I-protein
PAS	NN	O	I-protein
)	NN	O	I-protein
homology	NN	O	I-protein
region	NN	O	I-protein
.	NN	O	O

In	NN	O	O
extracts	NN	O	O
from	NN	O	O
nonstimulated	NN	O	O
cells	NN	O	O
the	NN	O	O
receptor	NN	O	O
is	NN	O	O
recovered	NN	O	O
in	NN	O	O
an	NN	O	O
inducible	NN	O	O
cytoplasmic	NN	O	O
form	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
90-kDa	NN	O	B-protein
heat	NN	O	I-protein
shock	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
hsp90	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
molecular	NN	O	B-protein
chaperone	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
reconstituted	NN	O	O
ligand-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
to	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
form	NN	O	I-protein
by	NN	O	O
using	NN	O	O
the	NN	O	O
dioxin	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
its	NN	O	O
bHLH-PAS	NN	O	B-protein
partner	NN	O	I-protein
factor	NN	O	I-protein
Arnt	NN	O	B-protein
expressed	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
translation	NN	O	O
in	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
the	NN	O	O
PAS	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
resulted	NN	O	O
in	NN	O	O
constitutive	NN	O	O
dimerization	NN	O	O
with	NN	O	O
Arnt	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
this	NN	O	O
receptor	NN	O	O
mutant	NN	O	O
showed	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
xenobiotic	NN	O	O
response	NN	O	O
element-binding	NN	O	O
activity	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
PAS	NN	O	B-protein
domain	NN	O	I-protein
may	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
DNA-binding	NN	O	O
affinity	NN	O	O
and/or	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
not	NN	O	O
possible	NN	O	O
to	NN	O	O
reconstitute	NN	O	O
dioxin	NN	O	B-protein
receptor	NN	O	I-protein
function	NN	O	O
with	NN	O	O
proteins	NN	O	O
expressed	NN	O	O
in	NN	O	O
wheat	NN	O	O
germ	NN	O	O
lysate	NN	O	O
.	NN	O	O

In	NN	O	O
line	NN	O	O
with	NN	O	O
these	NN	O	O
observations	NN	O	O
,	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
but	NN	O	O
not	NN	O	O
wheat	NN	O	O
germ	NN	O	O
lysate	NN	O	O
promoted	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
de	NN	O	O
novo	NN	O	O
synthesized	NN	O	O
dioxin	NN	O	B-protein
receptor	NN	O	I-protein
with	NN	O	O
hsp90	NN	O	B-protein
.	NN	O	O

At	NN	O	O
least	NN	O	O
two	NN	O	O
distinct	NN	O	O
domains	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
mediated	NN	O	O
interaction	NN	O	O
with	NN	O	O
hsp90	NN	O	B-protein
:	NN	O	O
the	NN	O	O
ligand-binding	NN	O	B-protein
domain	NN	O	I-protein
located	NN	O	O
within	NN	O	O
the	NN	O	O
PAS	NN	O	B-protein
region	NN	O	I-protein
and	NN	O	O
,	NN	O	O
surprisingly	NN	O	O
,	NN	O	O
the	NN	O	O
bHLH	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

Whereas	NN	O	O
ligand-binding	NN	O	O
activity	NN	O	O
correlated	NN	O	O
with	NN	O	O
association	NN	O	O
with	NN	O	O
hsp90	NN	O	B-protein
,	NN	O	O
bHLH-	NN	O	O
hsp90	NN	O	B-protein
interaction	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
but	NN	O	O
not	NN	O	O
for	NN	O	O
dimerization	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
.	NN	O	O

Several	NN	O	O
distinct	NN	O	O
roles	NN	O	O
for	NN	O	O
hsp90	NN	O	B-protein
in	NN	O	O
modulating	NN	O	O
dioxin	NN	O	B-protein
receptor	NN	O	I-protein
function	NN	O	O
are	NN	O	O
therefore	NN	O	O
likely	NN	O	O
:	NN	O	O
correct	NN	O	O
folding	NN	O	O
of	NN	O	O
the	NN	O	O
ligand-binding	NN	O	O
domain	NN	O	O
,	NN	O	O
interference	NN	O	O
with	NN	O	O
Arnt	NN	O	B-protein
heterodimerization	NN	O	O
,	NN	O	O
and	NN	O	O
folding	NN	O	O
of	NN	O	O
a	NN	O	O
DNA-binding	NN	O	O
conformation	NN	O	O
of	NN	O	O
the	NN	O	O
bHLH	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
dioxin	NN	O	B-protein
receptor	NN	O	I-protein
system	NN	O	O
provides	NN	O	O
a	NN	O	O
complex	NN	O	O
and	NN	O	O
interesting	NN	O	O
model	NN	O	O
of	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
by	NN	O	O
hsp90	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
aspirin-like	NN	O	O
drugs	NN	O	O
.	NN	O	O

Aspirin-like	NN	O	O
drugs	NN	O	O
(	NN	O	O
ALD	NN	O	O
)	NN	O	O
induce	NN	O	O
calcium	NN	O	O
mobilization	NN	O	O
,	NN	O	O
an	NN	O	O
essential	NN	O	O
component	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
but	NN	O	O
do	NN	O	O
not	NN	O	O
induce	NN	O	O
the	NN	O	O
biosynthesis	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
extent	NN	O	O
to	NN	O	O
which	NN	O	O
ALD	NN	O	O
may	NN	O	O
mimic	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
cytoplasmic	NN	O	O
and	NN	O	O
nuclear	NN	O	O
signaling	NN	O	O
steps	NN	O	O
in	NN	O	O
ALD-treated	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
ALD	NN	O	O
induce	NN	O	O
a	NN	O	O
transient	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
but	NN	O	O
have	NN	O	O
no	NN	O	O
effect	NN	O	O
(	NN	O	O
in	NN	O	O
comparison	NN	O	O
to	NN	O	O
anti-CD3	NN	O	B-protein
antibodies	NN	O	I-protein
)	NN	O	O
on	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
nor	NN	O	O
on	NN	O	O
PCL	NN	O	O
gamma	NN	O	O
1	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

ALD-induced	NN	O	O
calcium	NN	O	O
mobilization	NN	O	O
and	NN	O	O
PKC	NN	O	B-protein
activation	NN	O	O
are	NN	O	O
independent	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
activity	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
effect	NN	O	O
of	NN	O	O
herbimycin	NN	O	O
,	NN	O	O
a	NN	O	O
tyrosine-protein	NN	O	B-protein
kinase	NN	O	I-protein
-specific	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

Although	NN	O	O
we	NN	O	O
detected	NN	O	O
no	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
ALD-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
contain	NN	O	O
proteins	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
three	NN	O	O
regulatory	NN	O	B-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
:	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
binding	NN	O	O
activities	NN	O	O
are	NN	O	O
expressed	NN	O	O
only	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
depended	NN	O	O
on	NN	O	O
calcium	NN	O	O
mobilization	NN	O	O
and	NN	O	O
PKC	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
ALD	NN	O	O
cause	NN	O	O
transient	NN	O	O
but	NN	O	O
significant	NN	O	O
changes	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
transmembrane	NN	O	O
signaling	NN	O	O
,	NN	O	O
although	NN	O	O
some	NN	O	O
events	NN	O	O
induced	NN	O	O
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
antibodies	NN	O	I-protein
are	NN	O	O
not	NN	O	O
induced	NN	O	O
by	NN	O	O
ALD	NN	O	O
.	NN	O	O

The	NN	O	O
signal	NN	O	O
is	NN	O	O
transmitted	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
and	NN	O	O
induces	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
by	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
ALD	NN	O	O
stimulus	NN	O	O
is	NN	O	O
not	NN	O	O
capable	NN	O	O
of	NN	O	O
causing	NN	O	O
complete	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
is	NN	O	O
not	NN	O	O
a	NN	O	O
downstream	NN	O	O
effector	NN	O	O
of	NN	O	O
p21ras	NN	O	B-DNA
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
,	NN	O	O
downstream	NN	O	O
of	NN	O	O
p21ras	NN	O	B-DNA
,	NN	O	O
in	NN	O	O
activating	NN	O	O
interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
that	NN	O	O
PKC	NN	O	B-protein
is	NN	O	O
an	NN	O	O
effector	NN	O	B-protein
of	NN	O	O
p21ras	NN	O	B-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Data	NN	O	O
is	NN	O	O
presented	NN	O	O
,	NN	O	O
using	NN	O	O
the	NN	O	O
potent	NN	O	O
and	NN	O	O
selective	NN	O	O
PKC	NN	O	B-protein
inhibitor	NN	O	O
Ro	NN	O	O
31-8425	NN	O	O
and	NN	O	O
transient	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
constitutively	NN	O	O
active	NN	O	O
ras	NN	O	O
mutant	NN	O	O
,	NN	O	O
which	NN	O	O
clearly	NN	O	O
shows	NN	O	O
that	NN	O	O
PKC	NN	O	B-protein
is	NN	O	O
not	NN	O	O
downstream	NN	O	O
of	NN	O	O
p21ras	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Reporter	NN	O	O
gene	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcriptional	NN	O	O
activity	NN	O	O
is	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
expression	NN	O	O
of	NN	O	O
activated	NN	O	O
p21ras	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
involving	NN	O	O
PKC	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
calcium	NN	O	O
mobilization	NN	O	O
and	NN	O	O
ras	NN	O	B-protein
activation	NN	O	O
combine	NN	O	O
to	NN	O	O
provide	NN	O	O
the	NN	O	O
necessary	NN	O	O
components	NN	O	O
for	NN	O	O
production	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
C-terminus	NN	O	B-protein
of	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
activator	NN	O	I-protein
Oct-2	NN	O	B-protein
functions	NN	O	O
as	NN	O	O
an	NN	O	O
activation	NN	O	B-protein
domain	NN	O	I-protein
in	NN	O	O
yeast	NN	O	O
.	NN	O	O

Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
are	NN	O	O
human	NN	O	O
transcriptional	NN	O	B-protein
activators	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
DNA	NN	O	O
element	NN	O	O
but	NN	O	O
activate	NN	O	O
distinct	NN	O	O
sets	NN	O	O
of	NN	O	O
genes	NN	O	O
.	NN	O	O

We	NN	O	O
expressed	NN	O	O
these	NN	O	O
factors	NN	O	O
in	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
and	NN	O	O
observed	NN	O	O
greater	NN	O	O
than	NN	O	O
5-fold	NN	O	O
stimulation	NN	O	O
of	NN	O	O
a	NN	O	O
lacZ	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
only	NN	O	O
with	NN	O	O
Oct-2	NN	O	B-protein
.	NN	O	O

Transfer	NN	O	O
of	NN	O	O
the	NN	O	O
Oct-2	NN	O	B-protein
C-terminal	NN	O	I-protein
domain	NN	O	I-protein
onto	NN	O	O
either	NN	O	O
Oct-1	NN	O	B-protein
(	NN	O	O
Oct1.2	NN	O	B-protein
)	NN	O	O
or	NN	O	O
a	NN	O	O
nonactivating	NN	O	B-protein
DNA-binding	NN	O	I-protein
domain	NN	O	I-protein
from	NN	O	O
GAL4	NN	O	B-protein
created	NN	O	O
activators	NN	O	O
capable	NN	O	O
of	NN	O	O
greater	NN	O	O
than	NN	O	O
15	NN	O	O
and	NN	O	O
10-fold	NN	O	O
stimulation	NN	O	O
of	NN	O	O
activity	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
C-terminus	NN	O	B-protein
of	NN	O	O
Oct-2	NN	O	B-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
confer	NN	O	O
activation	NN	O	O
potential	NN	O	O
to	NN	O	O
nonactive	NN	O	O
DNA-binding	NN	O	B-DNA
fragments	NN	O	I-DNA
in	NN	O	O
yeast	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid-induced	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
glucocorticoids	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
earliest	NN	O	O
and	NN	O	O
most	NN	O	O
thoroughly	NN	O	O
studied	NN	O	O
models	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
exact	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
apoptosis	NN	O	O
occurs	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
is	NN	O	O
unknown	NN	O	O
many	NN	O	O
biochemical	NN	O	O
and	NN	O	O
molecular	NN	O	O
changes	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
occur	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
chromatin	NN	O	B-DNA
degradation	NN	O	O
and	NN	O	O
endonucleases	NN	O	B-protein
in	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
process	NN	O	O
has	NN	O	O
been	NN	O	O
closely	NN	O	O
studied	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
several	NN	O	O
oncogenes	NN	O	B-DNA
in	NN	O	O
glucocorticoid-induced	NN	O	O
cell	NN	O	O
lysis	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
clinical	NN	O	O
importance	NN	O	O
of	NN	O	O
glucocorticoid-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
lymphoid	NN	O	O
neoplasms	NN	O	O
has	NN	O	O
recently	NN	O	O
received	NN	O	O
increased	NN	O	O
attention	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin-2	NN	O	B-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
stat3	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

An	NN	O	O
early	NN	O	O
biochemical	NN	O	O
event	NN	O	O
associated	NN	O	O
with	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
through	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-2R	NN	O	B-protein
)	NN	O	O
is	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
several	NN	O	O
intracellular	NN	O	O
substrates	NN	O	O
.	NN	O	O

The	NN	O	O
exact	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
IL-2	NN	O	B-protein
regulates	NN	O	O
transcription	NN	O	O
of	NN	O	O
different	NN	O	O
genes	NN	O	O
is	NN	O	O
presently	NN	O	O
unknown	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
stimulation	NN	O	O
through	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-protein
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
subsequent	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
stat3	NN	O	B-protein
,	NN	O	O
a	NN	O	O
newly	NN	O	O
identified	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
proteins	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
stat1	NN	O	O
proteins	NN	O	O
were	NN	O	O
not	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
after	NN	O	O
IL-2	NN	O	B-protein
ligation	NN	O	O
,	NN	O	O
whereas	NN	O	O
tyrosine-phosphorylated	NN	O	B-protein
stat1	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
91	NN	O	B-protein
and	NN	O	I-protein
84	NN	O	I-protein
kDa	NN	O	I-protein
proteins	NN	O	I-protein
)	NN	O	O
were	NN	O	O
translocated	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
following	NN	O	O
interferon-gamma	NN	O	B-protein
treatment	NN	O	O
of	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Apart	NN	O	O
from	NN	O	O
stat3	NN	O	B-protein
,	NN	O	O
another	NN	O	O
cytoplasmic	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
subsequently	NN	O	O
translocated	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

This	NN	O	O
protein	NN	O	O
had	NN	O	O
an	NN	O	O
apparent	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
84	NN	O	O
kDa	NN	O	O
and	NN	O	O
was	NN	O	O
not	NN	O	O
recognized	NN	O	O
by	NN	O	O
stat3	NN	O	B-protein
or	NN	O	O
stat1	NN	O	B-protein
mAb	NN	O	I-protein
or	NN	O	O
antisera	NN	O	O
.	NN	O	O

Since	NN	O	O
IL-2	NN	O	B-protein
induced	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
84	NN	O	B-protein
kDa	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
stat3	NN	O	B-protein
followed	NN	O	O
identical	NN	O	O
kinetics	NN	O	O
,	NN	O	O
p84	NN	O	B-protein
is	NN	O	O
a	NN	O	O
candidate	NN	O	O
for	NN	O	O
a	NN	O	O
new	NN	O	O
,	NN	O	O
yet	NN	O	O
undefined	NN	O	O
,	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
STAT	NN	O	O
family	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
subsequent	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
stat3	NN	O	B-protein
and	NN	O	O
an	NN	O	O
as	NN	O	O
yet	NN	O	O
undefined	NN	O	O
84-kDa	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
antigen-specific	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
Tax	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
involves	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
RelA	NN	O	B-protein
(	NN	O	O
p65	NN	O	B-protein
)	NN	O	O
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
c-rel	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
tax	NN	O	O
gene	NN	O	O
product	NN	O	O
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
that	NN	O	O
both	NN	O	O
stimulates	NN	O	O
viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
activates	NN	O	O
an	NN	O	O
array	NN	O	O
of	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_type
growth	NN	O	O
.	NN	O	O

Tax	NN	O	O
acts	NN	O	O
indirectly	NN	O	O
by	NN	O	O
inducing	NN	O	O
or	NN	O	O
modifying	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
various	NN	O	O
host	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
enhancer-binding	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
many	NN	O	O
of	NN	O	O
these	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
factors	NN	O	I-protein
are	NN	O	O
sequestered	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
by	NN	O	O
various	NN	O	O
ankyrin-rich	NN	O	B-protein
inhibitory	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
including	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

HTLV-I	NN	O	B-protein
Tax	NN	O	I-protein
expression	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
constitutive	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
biologically	NN	O	O
active	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
c-Rel	NN	O	B-protein
complexes	NN	O	I-protein
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
biochemical	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
underlying	NN	O	O
this	NN	O	O
response	NN	O	O
remains	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
-stimulated	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
both	NN	O	O
HTLV-I-infected	NN	O	B-cell_line
and	NN	O	I-cell_line
Tax-transfected	NN	O	I-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
rapid	NN	O	O
proteolytic	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
prior	NN	O	O
in	NN	O	O
vitro	NN	O	O
studies	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
a	NN	O	O
fraction	NN	O	O
of	NN	O	O
the	NN	O	O
phosphorylated	NN	O	O
form	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
remains	NN	O	O
physically	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
but	NN	O	O
is	NN	O	O
subject	NN	O	O
to	NN	O	O
rapid	NN	O	O
degradation	NN	O	O
,	NN	O	O
thereby	NN	O	O
promoting	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
active	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
nuclear	NN	O	O
c-Rel	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
RelA	NN	O	B-protein
(	NN	O	O
p65	NN	O	B-protein
)	NN	O	O
subunit	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
activates	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
c-rel	NN	O	B-DNA
gene	NN	O	I-DNA
through	NN	O	O
an	NN	O	O
intrinsic	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
normal	NN	O	O
cells	NN	O	O
,	NN	O	O
the	NN	O	O
subsequent	NN	O	O
accumulation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
c-Rel	NN	O	I-protein
acts	NN	O	O
to	NN	O	O
inhibit	NN	O	O
its	NN	O	O
own	NN	O	O
continued	NN	O	O
production	NN	O	O
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
autoregulatory	NN	O	O
loop	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
multiple	NN	O	O
phosphorylations	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
in	NN	O	O
intact	NN	O	O
cells	NN	O	O
is	NN	O	O
mechanistically	NN	O	O
not	NN	O	O
well	NN	O	O
understood	NN	O	O
.	NN	O	O

Therefore	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
modifications	NN	O	O
imposed	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B/I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
components	NN	O	I-protein
following	NN	O	O
stimulation	NN	O	O
and	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
final	NN	O	O
step	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
induction	NN	O	O
in	NN	O	O
vivo	NN	O	O
involves	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
several	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
protein	NN	O	I-protein
families	NN	O	I-protein
.	NN	O	O

In	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
TNF-alpha	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
primarily	NN	O	O
of	NN	O	O
p50-p65	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
c-rel	NN	O	B-protein
.	NN	O	O

Both	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
precursors	NN	O	O
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
became	NN	O	O
strongly	NN	O	O
phosphorylated	NN	O	O
with	NN	O	O
the	NN	O	O
same	NN	O	O
kinetics	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
inducible	NN	O	O
phosphorylation	NN	O	O
after	NN	O	O
stimulation	NN	O	O
,	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
containing	NN	O	O
constitutive	NN	O	O
nuclear	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
revealed	NN	O	O
constitutively	NN	O	O
phosphorylated	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

Phosphorylation	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
induced	NN	O	O
processing	NN	O	O
of	NN	O	O
the	NN	O	O
precursors	NN	O	O
p100	NN	O	B-protein
and	NN	O	O
p105	NN	O	B-protein
and	NN	O	O
by	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

As	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
model	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
impedes	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
as	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
before	NN	O	O
for	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
even	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
c-rel	NN	O	B-protein
,	NN	O	O
was	NN	O	O
phosphorylated	NN	O	O
after	NN	O	O
induction	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
TNF-alpha	NN	O	B-protein
selectively	NN	O	O
activates	NN	O	O
only	NN	O	O
specific	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
heteromers	NN	O	O
and	NN	O	O
that	NN	O	O
modifications	NN	O	O
regulate	NN	O	O
not	NN	O	O
only	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
molecules	NN	O	I-protein
but	NN	O	O
also	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
molecules	NN	O	O
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
cellular	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
was	NN	O	O
phosphorylation-dependent	NN	O	O
and	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
p65-containing	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
was	NN	O	O
enhanced	NN	O	O
by	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
by	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
assumed	NN	O	O
to	NN	O	O
be	NN	O	O
triggered	NN	O	O
by	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
,	NN	O	O
also	NN	O	O
coincided	NN	O	O
with	NN	O	O
incorporation	NN	O	O
of	NN	O	O
phosphate	NN	O	O
into	NN	O	O
the	NN	O	O
same	NN	O	O
subunits	NN	O	O
that	NN	O	O
were	NN	O	O
modified	NN	O	O
after	NN	O	O
stimulation	NN	O	O
by	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
phosphorylation	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
general	NN	O	O
mechanism	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Treatment	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
various	NN	O	O
combinations	NN	O	O
of	NN	O	O
retinoids	NN	O	O
and	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25	NN	O	O
dihydroxyvitamin	NN	O	O
D3	NN	O	O
results	NN	O	O
in	NN	O	O
differentiation	NN	O	O
towards	NN	O	O
neutrophils	NN	O	B-cell_type
or	NN	O	O
monocytes	NN	O	B-cell_type
or	NN	O	O
a	NN	O	O
failure	NN	O	O
to	NN	O	O
differentiate	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
well	NN	O	O
documented	NN	O	O
that	NN	O	O
treatment	NN	O	O
of	NN	O	O
serum-grown	NN	O	B-cell_line
HL60	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
M	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
)	NN	O	O
induces	NN	O	O
neutrophil	NN	O	B-cell_type
differentiation	NN	O	O
,	NN	O	O
whereas	NN	O	O
treatment	NN	O	O
with	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
M	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25	NN	O	O
dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
D3	NN	O	O
)	NN	O	O
induces	NN	O	O
differentiation	NN	O	O
towards	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
recent	NN	O	O
investigations	NN	O	O
,	NN	O	O
using	NN	O	O
serum-free	NN	O	B-cell_line
grown	NN	O	I-cell_line
HL60	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
,	NN	O	O
at	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
M	NN	O	O
,	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
neutrophil	NN	O	O
differentiation	NN	O	O
and	NN	O	O
that	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
,	NN	O	O
at	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
M	NN	O	O
,	NN	O	O
reduced	NN	O	O
the	NN	O	O
D3	NN	O	O
concentration	NN	O	O
required	NN	O	O
for	NN	O	O
monocyte	NN	O	B-cell_type
differentiation	NN	O	O
to	NN	O	O
5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
co-operative	NN	O	O
interactions	NN	O	O
between	NN	O	O
all-trans	NN	O	O
and	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
and	NN	O	O
D3	NN	O	O
which	NN	O	O
promote	NN	O	O
neutrophil	NN	O	O
and	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
have	NN	O	O
been	NN	O	O
analysed	NN	O	O
in	NN	O	O
detail	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
serum-free	NN	O	B-cell_line
grown	NN	O	I-cell_line
HL60	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
M	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
or	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
resulted	NN	O	O
in	NN	O	O
sub-optimal	NN	O	O
neutrophil	NN	O	O
differentiation	NN	O	O
(	NN	O	O
up	NN	O	O
to	NN	O	O
25	NN	O	O
%	NN	O	O
mature	NN	O	O
cells	NN	O	O
)	NN	O	O
.	NN	O	O

As	NN	O	O
shown	NN	O	O
for	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
,	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
cooperated	NN	O	O
with	NN	O	O
D3	NN	O	O
to	NN	O	O
promote	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Culture	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
M	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
together	NN	O	O
with	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
concentrations	NN	O	O
of	NN	O	O
D3	NN	O	O
resulted	NN	O	O
in	NN	O	O
promotion	NN	O	O
of	NN	O	O
neutrophil	NN	O	O
differentiation	NN	O	O
at	NN	O	O
10	NN	O	O
(	NN	O	O
-15	NN	O	O
)	NN	O	O
-10	NN	O	O
(	NN	O	O
-12	NN	O	O
)	NN	O	O
D3	NN	O	O
,	NN	O	O
a	NN	O	O
failure	NN	O	O
to	NN	O	O
differentiate	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
at	NN	O	O
10	NN	O	O
(	NN	O	O
-11	NN	O	O
)	NN	O	O
-10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
M	NN	O	O
D3	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
co-operativity	NN	O	O
between	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
and	NN	O	O
5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M	NN	O	O
D3	NN	O	O
in	NN	O	O
inducing	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
neutrophil	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Similar	NN	O	O
results	NN	O	O
were	NN	O	O
obtained	NN	O	O
when	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
treated	NN	O	O
with	NN	O	O
5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
together	NN	O	O
with	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
concentrations	NN	O	O
of	NN	O	O
D3	NN	O	O
.	NN	O	O

Cross	NN	O	O
titration	NN	O	O
analyses	NN	O	O
of	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
and	NN	O	O
D3	NN	O	O
on	NN	O	O
HL60	NN	O	B-cell_line
cell	NN	O	I-cell_line
differentiation	NN	O	O
were	NN	O	O
undertaken	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
boundaries	NN	O	O
of	NN	O	O
the	NN	O	O
concentrations	NN	O	O
of	NN	O	O
each	NN	O	O
agent	NN	O	O
,	NN	O	O
alone	NN	O	O
and	NN	O	O
in	NN	O	O
combination	NN	O	O
,	NN	O	O
that	NN	O	O
give	NN	O	O
rise	NN	O	O
to	NN	O	O
optimal	NN	O	O
neutrophil	NN	O	O
and	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
observed	NN	O	O
cooperativities	NN	O	O
between	NN	O	O
either	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
or	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
and	NN	O	O
D3	NN	O	O
have	NN	O	O
important	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
combinations	NN	O	O
of	NN	O	O
these	NN	O	O
agents	NN	O	O
in	NN	O	O
differentiation	NN	O	O
therapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Isolation	NN	O	O
of	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
sequence	NN	O	O
tags	NN	O	O
from	NN	O	O
steroid-responsive	NN	O	B-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
mRNA	NN	O	B-RNA
differential	NN	O	O
display	NN	O	O
.	NN	O	O

Transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
steroid-regulated	NN	O	B-DNA
gene	NN	O	I-DNA
networks	NN	O	I-DNA
by	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
proteins	NN	O	I-protein
results	NN	O	O
in	NN	O	O
the	NN	O	O
coordinate	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
limited	NN	O	O
number	NN	O	O
of	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Although	NN	O	O
much	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
structure	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
steroid	NN	O	O
receptors	NN	O	O
,	NN	O	O
relatively	NN	O	O
few	NN	O	O
cell-specific	NN	O	O
steroid-regulated	NN	O	O
genes	NN	O	O
have	NN	O	O
been	NN	O	O
isolated	NN	O	O
and	NN	O	O
characterized	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
we	NN	O	O
describe	NN	O	O
results	NN	O	O
using	NN	O	O
mRNA	NN	O	B-RNA
differential	NN	O	O
display	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
PCR	NN	O	O
(	NN	O	O
DDPCR	NN	O	O
)	NN	O	O
to	NN	O	O
identify	NN	O	O
and	NN	O	O
isolate	NN	O	O
short	NN	O	O
cDNA	NN	O	O
sequence	NN	O	O
tags	NN	O	O
from	NN	O	O
thymocyte	NN	O	B-cell_type
and	NN	O	O
prostate	NN	O	B-cell_type
cells	NN	O	I-cell_type
under	NN	O	O
various	NN	O	O
hormone	NN	O	O
conditions	NN	O	O
.	NN	O	O

Using	NN	O	O
this	NN	O	O
technique	NN	O	O
we	NN	O	O
have	NN	O	O
isolated	NN	O	O
several	NN	O	O
differentially	NN	O	B-DNA
expressed	NN	O	I-DNA
sequence	NN	O	I-DNA
tags	NN	O	I-DNA
(	NN	O	O
DESTs	NN	O	B-DNA
)	NN	O	O
from	NN	O	O
the	NN	O	O
mouse	NN	O	B-cell_line
thymocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
WEHI	NN	O	B-cell_line
7.2	NN	O	I-cell_line
.	NN	O	O

Two	NN	O	O
of	NN	O	O
these	NN	O	O
DESTs	NN	O	B-DNA
,	NN	O	O
GIG10	NN	O	B-DNA
and	NN	O	O
GIG18	NN	O	B-DNA
,	NN	O	O
are	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
within	NN	O	O
2	NN	O	O
h	NN	O	O
of	NN	O	O
treatment	NN	O	O
.	NN	O	O

GIG10	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
novel	NN	O	O
sequence	NN	O	O
and	NN	O	O
GIG18	NN	O	B-DNA
is	NN	O	O
the	NN	O	O
mouse	NN	O	O
homologue	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
expressed	NN	O	I-DNA
sequence	NN	O	I-DNA
tag	NN	O	I-DNA
isolated	NN	O	O
from	NN	O	O
activated	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
also	NN	O	O
used	NN	O	O
DDPCR	NN	O	O
to	NN	O	O
isolate	NN	O	O
DESTs	NN	O	B-DNA
from	NN	O	O
androgen-modulated	NN	O	O
rat	NN	O	O
ventral	NN	O	O
prostate	NN	O	O
tissue	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
we	NN	O	O
characterized	NN	O	O
and	NN	O	O
found	NN	O	O
to	NN	O	O
correspond	NN	O	O
to	NN	O	O
the	NN	O	O
3	NN	O	B-RNA
'	NN	O	I-RNA
end	NN	O	I-RNA
of	NN	O	O
prostatic	NN	O	B-RNA
spermine	NN	O	I-RNA
binding	NN	O	I-RNA
protein	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
a	NN	O	O
known	NN	O	O
androgen-regulated	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Modifications	NN	O	O
of	NN	O	O
the	NN	O	O
original	NN	O	O
DDPCR	NN	O	O
protocol	NN	O	O
,	NN	O	O
which	NN	O	O
we	NN	O	O
found	NN	O	O
can	NN	O	O
potentially	NN	O	O
decrease	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
isolating	NN	O	O
false-positive	NN	O	O
DESTs	NN	O	B-DNA
,	NN	O	O
are	NN	O	O
described	NN	O	O
and	NN	O	O
the	NN	O	O
merits	NN	O	O
of	NN	O	O
DDPCR	NN	O	O
,	NN	O	O
relative	NN	O	O
to	NN	O	O
other	NN	O	O
differential	NN	O	O
cDNA	NN	O	O
cloning	NN	O	O
strategies	NN	O	O
,	NN	O	O
are	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
after	NN	O	O
infection	NN	O	O
by	NN	O	O
EBV	NN	O	O
.	NN	O	O

The	NN	O	O
discovery	NN	O	O
of	NN	O	O
EBV	NN	O	O
in	NN	O	O
certain	NN	O	O
T	NN	O	O
cell	NN	O	O
malignancies	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
CR2/CD21	NN	O	B-protein
,	NN	O	O
on	NN	O	O
a	NN	O	O
population	NN	O	O
of	NN	O	O
immature	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
,	NN	O	O
T	NN	O	B-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
childhood	NN	O	B-cell_type
acute	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphoblastic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
suggested	NN	O	O
that	NN	O	O
EBV-receptor	NN	O	O
interactions	NN	O	O
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
of	NN	O	O
importance	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
,	NN	O	O
within	NN	O	O
the	NN	O	O
thymus	NN	O	O
,	NN	O	O
a	NN	O	O
population	NN	O	O
of	NN	O	O
large	NN	O	B-cell_type
,	NN	O	I-cell_type
immature	NN	O	I-cell_type
cells	NN	O	I-cell_type
expresses	NN	O	O
CD21	NN	O	B-protein
.	NN	O	O

EBV	NN	O	O
altered	NN	O	O
the	NN	O	O
activation	NN	O	O
responses	NN	O	O
of	NN	O	O
immature	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Triggering	NN	O	O
through	NN	O	O
CD2	NN	O	B-protein
is	NN	O	O
mitogenic	NN	O	O
for	NN	O	O
mature	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
immature	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
during	NN	O	O
infection	NN	O	O
by	NN	O	O
EBV	NN	O	O
,	NN	O	O
ligation	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
caused	NN	O	O
thymocytes	NN	O	B-cell_type
to	NN	O	O
proliferate	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

This	NN	O	O
function	NN	O	O
was	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
EBV	NN	O	O
with	NN	O	O
its	NN	O	O
receptor	NN	O	O
,	NN	O	O
CD21	NN	O	B-protein
,	NN	O	O
but	NN	O	O
was	NN	O	O
caused	NN	O	O
by	NN	O	O
infection	NN	O	O
rather	NN	O	O
than	NN	O	O
surface	NN	O	O
signaling	NN	O	O
,	NN	O	O
because	NN	O	O
neither	NN	O	O
specific	NN	O	B-protein
mAb	NN	O	I-protein
nor	NN	O	O
the	NN	O	O
P3HR-1	NN	O	O
strain	NN	O	O
of	NN	O	O
virus	NN	O	O
mimicked	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
B95-8	NN	O	O
.	NN	O	O

Immature	NN	O	O
thymocytes	NN	O	B-cell_type
were	NN	O	O
infected	NN	O	O
by	NN	O	O
EBV	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
the	NN	O	O
internalization	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
genome	NN	O	I-DNA
and	NN	O	O
its	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
B95-8	NN	O	B-protein
,	NN	O	O
EBNA-2	NN	O	B-RNA
transcripts	NN	O	I-RNA
were	NN	O	O
identified	NN	O	O
within	NN	O	O
infected	NN	O	B-cell_line
thymocyte	NN	O	I-cell_line
populations	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	O
replicative	NN	O	O
pathway	NN	O	O
were	NN	O	O
expressed	NN	O	O
during	NN	O	O
infection	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
components	NN	O	O
included	NN	O	O
transcription	NN	O	O
of	NN	O	O
BZLF-1	NN	O	B-DNA
,	NN	O	O
an	NN	O	O
early	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
characterizes	NN	O	O
EBV-infected	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
after	NN	O	O
disruption	NN	O	O
of	NN	O	O
latency	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
transcript	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
encoding	NN	O	O
the	NN	O	O
recently	NN	O	O
characterized	NN	O	O
RAZ	NN	O	B-protein
,	NN	O	O
which	NN	O	O
also	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
replicative	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
consequences	NN	O	O
of	NN	O	O
EBV	NN	O	O
infection	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
an	NN	O	O
early	NN	O	O
stage	NN	O	O
of	NN	O	O
differentiation	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
failure	NN	O	O
of	NN	O	O
normal	NN	O	O
T	NN	O	O
cell	NN	O	O
repertoire	NN	O	O
development	NN	O	O
,	NN	O	O
autoimmunity	NN	O	O
,	NN	O	O
or	NN	O	O
malignancy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
intranasal	NN	O	O
glucocorticoids	NN	O	O
on	NN	O	O
endogenous	NN	O	O
glucocorticoid	NN	O	O
peripheral	NN	O	O
and	NN	O	O
central	NN	O	O
function	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
are	NN	O	O
among	NN	O	O
the	NN	O	O
most	NN	O	O
potent	NN	O	O
antiinflammatory	NN	O	O
agents	NN	O	O
that	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
rhinitis	NN	O	O
.	NN	O	O

Their	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
action	NN	O	O
are	NN	O	O
multiple	NN	O	O
and	NN	O	O
complex	NN	O	O
and	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
reports	NN	O	O
describe	NN	O	O
significant	NN	O	O
systemic	NN	O	O
effects	NN	O	O
of	NN	O	O
locally	NN	O	O
administered	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
evaluate	NN	O	O
the	NN	O	O
short-term	NN	O	O
systemic	NN	O	O
effects	NN	O	O
of	NN	O	O
intranasally	NN	O	O
administered	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
14	NN	O	O
normal	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
two	NN	O	O
doses	NN	O	O
of	NN	O	O
either	NN	O	O
budesonide	NN	O	O
(	NN	O	O
BUD	NN	O	O
)	NN	O	O
or	NN	O	O
fluticasone	NN	O	O
propionate	NN	O	O
(	NN	O	O
FP	NN	O	O
)	NN	O	O
for	NN	O	O
2	NN	O	O
weeks	NN	O	O
.	NN	O	O

Before	NN	O	O
treatment	NN	O	O
,	NN	O	O
at	NN	O	O
regular	NN	O	O
intervals	NN	O	O
during	NN	O	O
the	NN	O	O
treatment	NN	O	O
,	NN	O	O
1	NN	O	O
week	NN	O	O
and	NN	O	O
finally	NN	O	O
6	NN	O	O
weeks	NN	O	O
after	NN	O	O
termination	NN	O	O
of	NN	O	O
treatment	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
on	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
(	NN	O	O
GR	NN	O	O
)	NN	O	O
and	NN	O	O
methallothionein	NN	O	O
(	NN	O	O
MTIIa	NN	O	O
)	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
levels	NN	O	O
were	NN	O	O
examined	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
using	NN	O	O
a	NN	O	O
solution	NN	O	O
hybridization	NN	O	O
assay	NN	O	O
.	NN	O	O

Serum	NN	O	O
cortisol	NN	O	O
,	NN	O	O
osteocalcin	NN	O	O
and	NN	O	O
urinary	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
were	NN	O	O
also	NN	O	O
determined	NN	O	O
.	NN	O	O

An	NN	O	O
insulin	NN	O	O
tolerance	NN	O	O
test	NN	O	O
(	NN	O	O
ITT	NN	O	O
)	NN	O	O
was	NN	O	O
performed	NN	O	O
at	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
second	NN	O	O
week	NN	O	O
of	NN	O	O
treatment	NN	O	O
and	NN	O	O
at	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
6-week	NN	O	O
washout	NN	O	O
period	NN	O	O
with	NN	O	O
no	NN	O	O
statistically	NN	O	O
significant	NN	O	O
change	NN	O	O
in	NN	O	O
cortisol	NN	O	O
response	NN	O	O
.	NN	O	O

In	NN	O	O
peripheral	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
were	NN	O	O
significantly	NN	O	O
down-regulated	NN	O	O
.	NN	O	O

MTIIa	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
increased	NN	O	O
significantly	NN	O	O
.	NN	O	O

Serum	NN	O	O
osteocalcin	NN	O	B-protein
decreased	NN	O	O
significantly	NN	O	O
during	NN	O	O
treatment	NN	O	O
with	NN	O	O
both	NN	O	O
BUD	NN	O	O
and	NN	O	O
FP	NN	O	O
.	NN	O	O

Serum	NN	O	O
cortisol	NN	O	O
decreased	NN	O	O
after	NN	O	O
1	NN	O	O
week	NN	O	O
of	NN	O	O
treatment	NN	O	O
whereas	NN	O	O
urinary	NN	O	O
cortisol	NN	O	O
was	NN	O	O
not	NN	O	O
affected	NN	O	O
until	NN	O	O
the	NN	O	O
second	NN	O	O
week	NN	O	O
of	NN	O	O
treatment	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
intranasal	NN	O	O
glucocorticoids	NN	O	O
at	NN	O	O
clinically	NN	O	O
recommended	NN	O	O
doses	NN	O	O
have	NN	O	O
not	NN	O	O
only	NN	O	O
significant	NN	O	O
systemic	NN	O	O
effects	NN	O	O
on	NN	O	O
adrenal	NN	O	O
function	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
have	NN	O	O
an	NN	O	O
effect	NN	O	O
on	NN	O	O
specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
effects	NN	O	O
are	NN	O	O
receptor-dependent	NN	O	O
,	NN	O	O
reversible	NN	O	O
,	NN	O	O
and	NN	O	O
according	NN	O	O
to	NN	O	O
serum	NN	O	O
and	NN	O	O
urinary	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
and	NN	O	O
ITT	NN	O	O
,	NN	O	O
leave	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
function	NN	O	O
intact	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
these	NN	O	O
short-term	NN	O	O
systemic	NN	O	O
effects	NN	O	O
were	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
noticeable	NN	O	O
side-effects	NN	O	O
usually	NN	O	O
observed	NN	O	O
during	NN	O	O
long-term	NN	O	O
treatment	NN	O	O
with	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

-DOCSTART-	O

Biphasic	NN	O	O
control	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
:	NN	O	O
role	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
TcR	NN	O	I-protein
)	NN	O	I-protein
/CD3	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
primary	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
studied	NN	O	O
at	NN	O	O
various	NN	O	O
times	NN	O	O
after	NN	O	O
activation	NN	O	O
.	NN	O	O

Only	NN	O	O
p50	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
protein	NN	O	I-protein
bound	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
interleukin-2	NN	O	B-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
IL-2R	NN	O	I-DNA
)	NN	O	I-DNA
alpha	NN	O	I-DNA
chain	NN	O	I-DNA
promoter	NN	O	I-DNA
on	NN	O	O
resting	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
immediately	NN	O	O
after	NN	O	O
TcR/CD3	NN	O	B-protein
cross-linking	NN	O	O
(	NN	O	O
after	NN	O	O
approximately	NN	O	O
1	NN	O	O
h	NN	O	O
;	NN	O	O
immediate	NN	O	O
)	NN	O	O
binding	NN	O	O
of	NN	O	O
p50.p65	NN	O	B-protein
heterodimers	NN	O	I-protein
was	NN	O	O
observed	NN	O	O
.	NN	O	O

p50.c-rel	NN	O	B-protein
heterodimers	NN	O	I-protein
were	NN	O	O
also	NN	O	O
detected	NN	O	O
bound	NN	O	O
to	NN	O	O
this	NN	O	O
sequence	NN	O	O
at	NN	O	O
early	NN	O	O
time	NN	O	O
points	NN	O	O
(	NN	O	O
7-16	NN	O	O
h	NN	O	O
;	NN	O	O
early	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
both	NN	O	O
remained	NN	O	O
active	NN	O	O
at	NN	O	O
later	NN	O	O
time	NN	O	O
points	NN	O	O
(	NN	O	O
40	NN	O	O
h	NN	O	O
;	NN	O	O
late	NN	O	O
)	NN	O	O
after	NN	O	O
activation	NN	O	O
.	NN	O	O

This	NN	O	O
regulation	NN	O	O
takes	NN	O	O
place	NN	O	O
mainly	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
c-rel	NN	O	B-protein
,	NN	O	O
at	NN	O	O
immediate	NN	O	O
and	NN	O	O
early	NN	O	O
time	NN	O	O
points	NN	O	O
.	NN	O	O

Activation	NN	O	O
also	NN	O	O
induced	NN	O	O
c-rel	NN	O	O
and	NN	O	O
p105/p50	NN	O	O
mRNA	NN	O	O
synthesis	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
p65	NN	O	B-RNA
mRNA	NN	O	I-RNA
whose	NN	O	O
expression	NN	O	O
was	NN	O	O
constitutive	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
all	NN	O	O
those	NN	O	O
early	NN	O	O
and	NN	O	O
late	NN	O	O
events	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
immediate	NN	O	O
ones	NN	O	O
,	NN	O	O
were	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
neutralizing	NN	O	O
anti-tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	I-protein
TNF-alpha	NN	O	I-protein
)	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
cycloheximide	NN	O	O
prevented	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
c-rel	NN	O	B-protein
translocation	NN	O	O
and	NN	O	O
consequent	NN	O	O
formation	NN	O	O
of	NN	O	O
active	NN	O	O
binding	NN	O	O
heterodimers	NN	O	O
,	NN	O	O
at	NN	O	O
early	NN	O	O
and	NN	O	O
late	NN	O	O
times	NN	O	O
.	NN	O	O

Cyclosporin	NN	O	O
A	NN	O	O
impaired	NN	O	O
not	NN	O	O
only	NN	O	O
early	NN	O	O
and	NN	O	O
late	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
immediate	NN	O	O
events	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
prevented	NN	O	O
all	NN	O	O
inhibition	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
by	NN	O	O
TcR/CD3	NN	O	B-protein
signals	NN	O	O
is	NN	O	O
biphasic	NN	O	O
:	NN	O	O
TcR/CD3	NN	O	B-protein
triggers	NN	O	O
its	NN	O	O
immediate	NN	O	O
translocation	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
transient	NN	O	O
if	NN	O	O
no	NN	O	O
TNF-alpha	NN	O	B-protein
is	NN	O	O
present	NN	O	O
.	NN	O	O

TNF-alpha	NN	O	B-protein
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
emerges	NN	O	O
as	NN	O	O
the	NN	O	O
main	NN	O	O
factor	NN	O	O
responsible	NN	O	O
for	NN	O	O
a	NN	O	O
second	NN	O	O
phase	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
regulation	NN	O	O
,	NN	O	O
controlling	NN	O	O
both	NN	O	O
translocation	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
c-rel	NN	O	B-protein
,	NN	O	O
and	NN	O	O
new	NN	O	O
mRNA	NN	O	B-RNA
synthesis	NN	O	O
for	NN	O	O
c-rel	NN	O	B-protein
and	NN	O	O
p105/p50	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Sp1	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
cis-acting	NN	O	B-DNA
region	NN	O	I-DNA
,	NN	O	O
GM-kappa	NN	O	B-DNA
B/GC-box	NN	O	I-DNA
(	NN	O	O
positions	NN	O	B-DNA
-95	NN	O	I-DNA
and	NN	O	O
-73	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
within	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
required	NN	O	O
for	NN	O	O
maximal	NN	O	O
induction	NN	O	O
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phorbol-12-myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
(	NN	O	O
A23187	NN	O	O
)	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

GM-kappa	NN	O	B-protein
B	NN	O	I-protein
defines	NN	O	O
a	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
GC-box	NN	O	B-DNA
defines	NN	O	O
a	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
three	NN	O	O
(	NN	O	O
A1	NN	O	B-protein
,	NN	O	O
A2	NN	O	B-protein
,	NN	O	O
B	NN	O	B-protein
)	NN	O	O
constitutive	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
three	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
GC-box	NN	O	B-DNA
can	NN	O	O
functionally	NN	O	O
compensate	NN	O	O
for	NN	O	O
the	NN	O	O
GM-kappa	NN	O	B-DNA
B/GC-box	NN	O	I-DNA
region	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
GC-motif	NN	O	O
can	NN	O	O
function	NN	O	O
independently	NN	O	O
of	NN	O	O
the	NN	O	O
GM-kappa	NN	O	B-DNA
B	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
major	NN	O	O
GC-box	NN	O	B-DNA
binding	NN	O	O
activity	NN	O	O
A1	NN	O	B-protein
was	NN	O	O
purified	NN	O	O
and	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Sp1	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
depletion	NN	O	O
of	NN	O	O
Sp1	NN	O	B-protein
(	NN	O	O
A1	NN	O	B-protein
)	NN	O	O
from	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
specifically	NN	O	O
decreases	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
activity	NN	O	O
on	NN	O	O
GM-CSF	NN	O	B-protein
templates	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
promoter	NN	O	I-DNA
has	NN	O	O
a	NN	O	O
base	NN	O	O
difference	NN	O	O
within	NN	O	O
the	NN	O	O
GC-box	NN	O	B-DNA
,	NN	O	O
we	NN	O	O
speculate	NN	O	O
that	NN	O	O
this	NN	O	O
may	NN	O	O
explain	NN	O	O
why	NN	O	O
the	NN	O	O
human	NN	O	O
promoter	NN	O	O
is	NN	O	O
weak	NN	O	O
and	NN	O	O
that	NN	O	O
an	NN	O	O
upstream	NN	O	B-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
factor	NN	O	O
that	NN	O	O
regulates	NN	O	O
the	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
major	NN	O	I-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
gene	NN	O	I-DNA
DPA	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
subfamily	NN	O	O
of	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
proteins	NN	O	I-protein
that	NN	O	O
includes	NN	O	O
a	NN	O	O
Drosophila	NN	O	B-protein
developmental	NN	O	I-protein
control	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

A	NN	O	O
novel	NN	O	O
DNA	NN	O	B-DNA
sequence	NN	O	I-DNA
element	NN	O	I-DNA
termed	NN	O	O
the	NN	O	O
J	NN	O	B-DNA
element	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulated	NN	O	O
expression	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
major	NN	O	I-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
genes	NN	O	I-DNA
was	NN	O	O
recently	NN	O	O
described	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
this	NN	O	O
element	NN	O	O
and	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
regulation	NN	O	O
further	NN	O	O
,	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
library	NN	O	I-DNA
was	NN	O	O
screened	NN	O	O
with	NN	O	O
oligonucleotide	NN	O	O
probes	NN	O	O
containing	NN	O	O
both	NN	O	O
the	NN	O	O
S	NN	O	B-DNA
element	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
nearby	NN	O	O
J	NN	O	B-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
DPA	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Several	NN	O	O
DNA	NN	O	B-DNA
clones	NN	O	I-DNA
were	NN	O	O
obtained	NN	O	O
by	NN	O	O
this	NN	O	O
procedure	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
,	NN	O	O
clone	NN	O	B-DNA
18	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
reported	NN	O	O
and	NN	O	O
characterized	NN	O	O
here	NN	O	O
.	NN	O	O

It	NN	O	O
encodes	NN	O	O
a	NN	O	O
protein	NN	O	O
predicted	NN	O	O
to	NN	O	O
contain	NN	O	O
688	NN	O	O
amino	NN	O	O
acid	NN	O	O
residues	NN	O	O
,	NN	O	O
including	NN	O	O
11	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
motifs	NN	O	I-protein
of	NN	O	O
the	NN	O	O
C2H2	NN	O	B-protein
type	NN	O	I-protein
in	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
region	NN	O	I-protein
,	NN	O	O
that	NN	O	O
are	NN	O	O
Kruppel-like	NN	O	O
in	NN	O	O
the	NN	O	O
conservation	NN	O	O
of	NN	O	O
the	NN	O	O
H/C	NN	O	B-protein
link	NN	O	I-protein
sequence	NN	O	I-protein
connecting	NN	O	O
them	NN	O	O
.	NN	O	O

The	NN	O	O
160	NN	O	B-protein
N-terminal	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nonfinger	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
clone	NN	O	B-protein
18	NN	O	I-protein
are	NN	O	O
highly	NN	O	O
homologous	NN	O	O
with	NN	O	O
similar	NN	O	O
regions	NN	O	O
of	NN	O	O
several	NN	O	O
other	NN	O	O
human	NN	O	B-DNA
,	NN	O	I-DNA
mouse	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
Drosophila	NN	O	I-DNA
sequences	NN	O	I-DNA
,	NN	O	O
defining	NN	O	O
a	NN	O	O
subfamily	NN	O	O
of	NN	O	O
Kruppel-like	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
proteins	NN	O	I-protein
termed	NN	O	O
TAB	NN	O	B-protein
(	NN	O	O
tramtrack	NN	O	B-protein
[	NN	O	I-protein
ttk	NN	O	I-protein
]	NN	O	I-protein
-associated	NN	O	I-protein
box	NN	O	I-protein
)	NN	O	O
here	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-DNA
sequences	NN	O	I-DNA
,	NN	O	O
ttk	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
a	NN	O	O
developmental	NN	O	B-DNA
control	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
while	NN	O	O
a	NN	O	O
second	NN	O	O
does	NN	O	O
not	NN	O	O
contain	NN	O	O
a	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
region	NN	O	I-protein
but	NN	O	O
encodes	NN	O	O
a	NN	O	O
structure	NN	O	O
important	NN	O	O
in	NN	O	O
oocyte	NN	O	O
development	NN	O	O
.	NN	O	O

An	NN	O	O
acidic	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
is	NN	O	O
located	NN	O	O
between	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
conserved	NN	O	I-protein
region	NN	O	I-protein
of	NN	O	O
clone	NN	O	B-protein
18	NN	O	I-protein
and	NN	O	O
its	NN	O	O
zinc	NN	O	B-protein
fingers	NN	O	I-protein
.	NN	O	O

This	NN	O	O
protein	NN	O	O
appears	NN	O	O
to	NN	O	O
require	NN	O	O
both	NN	O	O
the	NN	O	O
S	NN	O	B-DNA
and	NN	O	I-DNA
J	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
are	NN	O	O
separated	NN	O	O
by	NN	O	O
10	NN	O	O
bp	NN	O	O
for	NN	O	O
optimal	NN	O	O
binding	NN	O	O
.	NN	O	O

Antisense	NN	O	O
cDNA	NN	O	O
to	NN	O	O
clone	NN	O	B-protein
18	NN	O	I-protein
inhibited	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	O
construct	NN	O	O
containing	NN	O	O
the	NN	O	O
DPA	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
indicating	NN	O	O
its	NN	O	O
functional	NN	O	O
importance	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Separation	NN	O	O
of	NN	O	O
oxidant-initiated	NN	O	O
and	NN	O	O
redox-regulated	NN	O	O
steps	NN	O	O
in	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
.	NN	O	O

Studies	NN	O	O
presented	NN	O	O
here	NN	O	O
show	NN	O	O
that	NN	O	O
overall	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
signal	NN	O	O
transduction	NN	O	O
begins	NN	O	O
with	NN	O	O
a	NN	O	O
parallel	NN	O	O
series	NN	O	O
of	NN	O	O
stimuli-specific	NN	O	O
pathways	NN	O	O
through	NN	O	O
which	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	O
,	NN	O	O
oxidants	NN	O	O
(	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
and	NN	O	O
mitomycin	NN	O	O
C	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
(	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
)	NN	O	O
individually	NN	O	O
initiate	NN	O	O
signaling	NN	O	O
.	NN	O	O

These	NN	O	O
initial	NN	O	O
pathways	NN	O	O
culminate	NN	O	O
in	NN	O	O
a	NN	O	O
common	NN	O	O
pathway	NN	O	O
through	NN	O	O
which	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
stimulating	NN	O	O
agents	NN	O	O
ultimately	NN	O	O
signal	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

We	NN	O	O
distinguish	NN	O	O
the	NN	O	O
stimuli-specific	NN	O	O
pathways	NN	O	O
by	NN	O	O
showing	NN	O	O
that	NN	O	O
the	NN	O	O
oxidative	NN	O	O
stimuli	NN	O	O
trigger	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
only	NN	O	O
one	NN	O	O
of	NN	O	O
two	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
Wurzburg	NN	O	B-cell_line
but	NN	O	O
not	NN	O	O
Jurkat	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
readily	NN	O	O
stimulate	NN	O	O
in	NN	O	O
both	NN	O	O
lines	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
the	NN	O	O
common	NN	O	O
pathway	NN	O	O
as	NN	O	O
the	NN	O	O
simplest	NN	O	O
way	NN	O	O
of	NN	O	O
accounting	NN	O	O
for	NN	O	O
the	NN	O	O
common	NN	O	O
requirements	NN	O	O
and	NN	O	O
properties	NN	O	O
of	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
include	NN	O	O
a	NN	O	O
redox-regulatory	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
in	NN	O	O
this	NN	O	O
common	NN	O	O
pathway	NN	O	O
to	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
redox	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
in	NN	O	O
which	NN	O	O
oxidants	NN	O	O
do	NN	O	O
n't	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
;	NN	O	O
we	NN	O	O
put	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
the	NN	O	O
common	NN	O	O
pathway	NN	O	O
by	NN	O	O
showing	NN	O	O
that	NN	O	O
kinase	NN	O	O
activity	NN	O	O
(	NN	O	O
inhibitable	NN	O	O
by	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
and	NN	O	O
tyrphostin	NN	O	O
47	NN	O	O
)	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
all	NN	O	O
stimuli	NN	O	O
tested	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
lines	NN	O	O
.	NN	O	O

Since	NN	O	O
internal	NN	O	O
sites	NN	O	O
of	NN	O	O
oxidant	NN	O	O
production	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
-stimulated	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
since	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
and	NN	O	O
phosphatase	NN	O	O
activities	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
altered	NN	O	O
by	NN	O	O
oxidants	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
intracellular	NN	O	O
redox	NN	O	O
status	NN	O	O
controls	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
regulating	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
event	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
within	NN	O	O
the	NN	O	O
common	NN	O	O
step	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Deleted	NN	O	O
chromosome	NN	O	B-DNA
20	NN	O	I-DNA
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
Alagille	NN	O	O
syndrome	NN	O	O
isolated	NN	O	O
in	NN	O	O
a	NN	O	O
cell	NN	O	O
hybrid	NN	O	O
through	NN	O	O
leucine	NN	O	O
transport	NN	O	O
selection	NN	O	O
:	NN	O	O
study	NN	O	O
of	NN	O	O
three	NN	O	O
candidate	NN	O	O
genes	NN	O	O
.	NN	O	O

Alagille	NN	O	O
syndrome	NN	O	O
(	NN	O	O
AGS	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
well-defined	NN	O	O
genetic	NN	O	O
entity	NN	O	O
assigned	NN	O	O
to	NN	O	O
the	NN	O	O
short	NN	O	B-DNA
arm	NN	O	I-DNA
of	NN	O	O
Chromosome	NN	O	B-DNA
(	NN	O	I-DNA
Chr	NN	O	I-DNA
)	NN	O	I-DNA
20	NN	O	I-DNA
by	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
observations	NN	O	O
of	NN	O	O
AGS	NN	O	O
patients	NN	O	O
associated	NN	O	O
with	NN	O	O
microdeletions	NN	O	O
in	NN	O	O
this	NN	O	O
region	NN	O	O
.	NN	O	O

By	NN	O	O
fusing	NN	O	O
lymphoblastoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
an	NN	O	O
AGS	NN	O	O
patient	NN	O	O
that	NN	O	O
exhibited	NN	O	O
a	NN	O	O
microdeletion	NN	O	O
in	NN	O	O
the	NN	O	O
short	NN	O	B-DNA
arm	NN	O	I-DNA
of	NN	O	I-DNA
Chr	NN	O	I-DNA
20	NN	O	I-DNA
encompassing	NN	O	O
bands	NN	O	O
p11.23	NN	O	B-DNA
to	NN	O	I-DNA
p12.3	NN	O	I-DNA
with	NN	O	O
rodent	NN	O	B-cell_line
thermosensitive	NN	O	I-cell_line
mutant	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
CHOtsH1-1	NN	O	B-cell_line
)	NN	O	O
deficient	NN	O	O
in-leucyl-tRNA	NN	O	B-protein
synthetase	NN	O	I-protein
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
a	NN	O	O
somatic	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybrid	NN	O	I-cell_line
segregating	NN	O	O
the	NN	O	O
deleted	NN	O	O
human	NN	O	O
Chr	NN	O	B-DNA
20	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
hybrid	NN	O	O
clone	NN	O	O
,	NN	O	O
designated	NN	O	O
NR2	NN	O	B-cell_line
,	NN	O	O
was	NN	O	O
characterized	NN	O	O
by	NN	O	O
several	NN	O	O
methods	NN	O	O
,	NN	O	O
including	NN	O	O
PCR	NN	O	O
,	NN	O	O
with	NN	O	O
eight	NN	O	O
pairs	NN	O	O
of	NN	O	O
oligonucleotides	NN	O	O
mapped	NN	O	O
to	NN	O	O
Chr	NN	O	B-DNA
20	NN	O	I-DNA
:	NN	O	O
D20S5	NN	O	B-DNA
,	NN	O	O
D20S41	NN	O	B-DNA
,	NN	O	O
D20S42	NN	O	B-DNA
,	NN	O	O
D20S56	NN	O	B-DNA
,	NN	O	O
D20S57	NN	O	B-DNA
,	NN	O	O
D20S58	NN	O	B-DNA
,	NN	O	O
adenosine	NN	O	B-DNA
deaminase	NN	O	I-DNA
(	NN	O	O
ADA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
Prion	NN	O	B-DNA
protein	NN	O	I-DNA
(	NN	O	O
PRIP	NN	O	B-protein
)	NN	O	O
;	NN	O	O
Restriction	NN	O	O
Fragment	NN	O	O
Length	NN	O	O
Polymorphism	NN	O	O
(	NN	O	O
RFLP	NN	O	B-DNA
)	NN	O	O
analyses	NN	O	O
with	NN	O	O
four	NN	O	O
genomic	NN	O	B-DNA
anonymous	NN	O	I-DNA
probes	NN	O	I-DNA
(	NN	O	O
D20S5	NN	O	B-DNA
,	NN	O	O
cD3H12	NN	O	B-DNA
,	NN	O	O
D20S17	NN	O	B-DNA
,	NN	O	O
D20S18	NN	O	B-DNA
)	NN	O	O
;	NN	O	O
and	NN	O	O
fluorescent	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
with	NN	O	O
total	NN	O	B-DNA
human	NN	O	I-DNA
DNA	NN	O	I-DNA
and	NN	O	O
D20Z1	NN	O	B-DNA
,	NN	O	O
a	NN	O	O
sequence	NN	O	O
specific	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
Chr	NN	O	I-DNA
20	NN	O	I-DNA
centromere	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
probes	NN	O	O
.	NN	O	O

The	NN	O	O
NR2	NN	O	B-cell_line
hybrid	NN	O	I-cell_line
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
exclude	NN	O	O
three	NN	O	O
candidate	NN	O	B-DNA
genes	NN	O	I-DNA
for	NN	O	O
AGS	NN	O	O
:	NN	O	O
hepatic	NN	O	B-DNA
nuclear	NN	O	I-DNA
factor	NN	O	I-DNA
3	NN	O	I-DNA
beta	NN	O	I-DNA
(	NN	O	O
HNF3	NN	O	B-DNA
beta	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
paired	NN	O	B-DNA
box	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	O
PAX1	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
and	NN	O	O
cystatin	NN	O	B-DNA
C	NN	O	I-DNA
(	NN	O	O
CST3	NN	O	B-DNA
)	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
their	NN	O	O
localization	NN	O	O
outside	NN	O	O
of	NN	O	O
the	NN	O	O
deletion	NN	O	O
.	NN	O	O

The	NN	O	O
NR2	NN	O	B-cell_line
hybrid	NN	O	I-cell_line
is	NN	O	O
a	NN	O	O
powerful	NN	O	O
tool	NN	O	O
for	NN	O	O
the	NN	O	O
mapping	NN	O	O
of	NN	O	O
new	NN	O	O
probes	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
for	NN	O	O
obtaining	NN	O	O
new	NN	O	O
informative	NN	O	O
probes	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
deletion	NN	O	O
by	NN	O	O
subtractive	NN	O	O
cloning	NN	O	O
of	NN	O	O
the	NN	O	O
region	NN	O	O
.	NN	O	O

Such	NN	O	O
markers	NN	O	O
will	NN	O	O
be	NN	O	O
useful	NN	O	O
for	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
and	NN	O	O
screening	NN	O	O
of	NN	O	O
cDNA	NN	O	B-DNA
libraries	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Positive	NN	O	O
and	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
composite	NN	O	B-DNA
octamer	NN	O	I-DNA
motif	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
enhancer	NN	O	I-DNA
by	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
differentiating	NN	O	O
agent	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
previously	NN	O	O
reported	NN	O	O
to	NN	O	O
interfere	NN	O	O
with	NN	O	O
12-O-tetradecanoyl-phorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
/Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-induced	NN	O	O
signals	NN	O	O
for	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
-96	NN	O	B-DNA
to	NN	O	I-DNA
-66-bp	NN	O	I-DNA
octamer	NN	O	I-DNA
motif	NN	O	I-DNA
found	NN	O	O
in	NN	O	O
the	NN	O	O
enhancer	NN	O	O
for	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
(	NN	O	I-DNA
IL	NN	O	I-DNA
)	NN	O	I-DNA
-2	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
a	NN	O	O
major	NN	O	O
T	NN	O	B-protein
lymphocyte	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
IL-2	NN	O	B-DNA
octamer	NN	O	I-DNA
motif	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
composite	NN	O	O
cis-element	NN	O	B-DNA
which	NN	O	O
binds	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
TPA/Ca	NN	O	B-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-inducible	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
previously	NN	O	O
termed	NN	O	O
octamer-associated	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
OAP40	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
despite	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
active	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
domain	NN	O	O
,	NN	O	O
requires	NN	O	O
TPA/Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-induced	NN	O	O
signals	NN	O	O
to	NN	O	O
strongly	NN	O	O
transactivate	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
octamer	NN	O	I-DNA
motif	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

This	NN	O	O
Oct-2	NN	O	B-protein
-dependent	NN	O	O
transactivation	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
RA	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
intact	NN	O	B-protein
COOH-terminal	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
Oct-2	NN	O	B-protein
contributes	NN	O	O
to	NN	O	O
both	NN	O	O
TPA/Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-induced	NN	O	O
transactivation	NN	O	O
and	NN	O	O
the	NN	O	O
RA-mediated	NN	O	O
repression	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
both	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
components	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
factors	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
OAP40	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
transfected	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	I-DNA
jun-B	NN	O	I-DNA
,	NN	O	I-DNA
jun-D	NN	O	I-DNA
,	NN	O	I-DNA
c-fos	NN	O	I-DNA
,	NN	O	I-DNA
or	NN	O	I-DNA
Fos-B	NN	O	I-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
partially	NN	O	O
substitute	NN	O	O
for	NN	O	O
TPA	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
and	NN	O	O
cooperate	NN	O	O
with	NN	O	O
Oct-2	NN	O	B-protein
for	NN	O	O
the	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
combined	NN	O	O
OAP/octamer	NN	O	B-DNA
cis-element	NN	O	I-DNA
.	NN	O	O

Mutations	NN	O	O
of	NN	O	O
the	NN	O	O
genuine	NN	O	B-DNA
octamer-binding	NN	O	I-DNA
site	NN	O	I-DNA
abrogate	NN	O	O
both	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
and	NN	O	O
the	NN	O	O
TPA/Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-induced	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
OAP/octamer	NN	O	B-DNA
motif	NN	O	I-DNA
.	NN	O	O

OAP	NN	O	B-protein
confers	NN	O	O
to	NN	O	O
Oct-2	NN	O	B-protein
responsivity	NN	O	O
to	NN	O	O
both	NN	O	O
TPA/Ca2+	NN	O	O
and	NN	O	O
RA	NN	O	O
,	NN	O	O
since	NN	O	O
specific	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1/OAP-binding	NN	O	B-DNA
site	NN	O	I-DNA
significantly	NN	O	O
reduce	NN	O	O
the	NN	O	O
transactivation	NN	O	O
by	NN	O	O
Oct-2	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
TPA	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
and	NN	O	O
abolish	NN	O	O
the	NN	O	O
inhibition	NN	O	O
by	NN	O	O
RA	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
)	NN	O	O
alpha	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
inhibit	NN	O	O
in	NN	O	O
vitro	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
complex	NN	O	O
between	NN	O	O
the	NN	O	O
nuclear	NN	O	O
AP-1	NN	O	B-protein
/OAP	NN	O	B-protein
and	NN	O	O
its	NN	O	O
specific	NN	O	O
binding	NN	O	O
site	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
interference	NN	O	O
with	NN	O	O
Oct-2	NN	O	B-protein
-dependent	NN	O	O
cis-regulatory	NN	O	O
function	NN	O	O
of	NN	O	O
this	NN	O	O
AP-1	NN	O	B-protein
element	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
TPA/calcium-activated	NN	O	O
AP-1	NN	O	B-protein
/OAP	NN	O	B-protein
element	NN	O	O
is	NN	O	O
the	NN	O	O
main	NN	O	O
target	NN	O	O
of	NN	O	O
positive	NN	O	O
or	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
signals	NN	O	O
influencing	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
octamer	NN	O	I-DNA
motif	NN	O	I-DNA
,	NN	O	O
through	NN	O	O
synergism	NN	O	O
with	NN	O	O
Oct-2	NN	O	B-protein
and	NN	O	O
antagonism	NN	O	O
by	NN	O	O
RAR	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
exon	NN	O	B-DNA
7	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
common	NN	O	B-DNA
gamma	NN	O	I-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
causes	NN	O	O
a	NN	O	O
moderate	NN	O	O
form	NN	O	O
of	NN	O	O
X-linked	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
.	NN	O	O

Clinical	NN	O	O
and	NN	O	O
immunologic	NN	O	O
features	NN	O	O
of	NN	O	O
a	NN	O	O
recently	NN	O	O
recognized	NN	O	O
X-linked	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
disease	NN	O	O
(	NN	O	O
XCID	NN	O	O
)	NN	O	O
suggested	NN	O	O
that	NN	O	O
XCID	NN	O	O
and	NN	O	O
X-linked	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
(	NN	O	O
XSCID	NN	O	O
)	NN	O	O
might	NN	O	O
arise	NN	O	O
from	NN	O	O
different	NN	O	O
genetic	NN	O	O
defects	NN	O	O
.	NN	O	O

The	NN	O	O
recent	NN	O	O
discovery	NN	O	O
of	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
common	NN	O	B-DNA
gamma	NN	O	I-DNA
chain	NN	O	I-DNA
(	NN	O	I-DNA
gamma	NN	O	I-DNA
c	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
constituent	NN	O	O
of	NN	O	O
several	NN	O	O
cytokine	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
in	NN	O	O
XSCID	NN	O	O
provided	NN	O	O
an	NN	O	O
opportunity	NN	O	O
to	NN	O	O
test	NN	O	O
directly	NN	O	O
whether	NN	O	O
a	NN	O	O
previously	NN	O	O
unrecognized	NN	O	O
mutation	NN	O	O
in	NN	O	O
this	NN	O	O
same	NN	O	O
gene	NN	O	O
was	NN	O	O
responsible	NN	O	O
for	NN	O	O
XCID	NN	O	O
.	NN	O	O

The	NN	O	O
status	NN	O	O
of	NN	O	O
X	NN	O	B-DNA
chromosome	NN	O	I-DNA
inactivation	NN	O	O
in	NN	O	O
blood	NN	O	O
leukocytes	NN	O	O
from	NN	O	O
obligate	NN	O	O
carriers	NN	O	O
of	NN	O	O
XCID	NN	O	O
was	NN	O	O
determined	NN	O	O
from	NN	O	O
the	NN	O	O
polymorphic	NN	O	O
,	NN	O	O
short	NN	O	O
tandem	NN	O	O
repeats	NN	O	O
(	NN	O	O
CAG	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
also	NN	O	O
contains	NN	O	O
a	NN	O	O
methylation-sensitive	NN	O	B-DNA
HpaII	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

As	NN	O	O
in	NN	O	O
XSCID	NN	O	O
,	NN	O	O
X-chromosome	NN	O	O
inactivation	NN	O	O
in	NN	O	O
obligate	NN	O	O
carriers	NN	O	O
of	NN	O	O
XCID	NN	O	O
was	NN	O	O
nonrandom	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
X	NN	O	B-DNA
chromosome	NN	O	I-DNA
inactivation	NN	O	O
in	NN	O	O
PMNs	NN	O	O
was	NN	O	O
variable	NN	O	O
.	NN	O	O

Findings	NN	O	O
from	NN	O	O
this	NN	O	O
analysis	NN	O	O
prompted	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
c	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
this	NN	O	O
pedigree	NN	O	O
.	NN	O	O

A	NN	O	O
missense	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
region	NN	O	O
coding	NN	O	O
for	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
c	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
found	NN	O	O
in	NN	O	O
three	NN	O	O
affected	NN	O	O
males	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
a	NN	O	O
normal	NN	O	O
brother	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
this	NN	O	O
point	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
c	NN	O	I-DNA
gene	NN	O	I-DNA
leads	NN	O	O
to	NN	O	O
a	NN	O	O
less	NN	O	O
severe	NN	O	O
degree	NN	O	O
of	NN	O	O
deficiency	NN	O	O
in	NN	O	O
cellular	NN	O	O
and	NN	O	O
humoral	NN	O	O
immunity	NN	O	O
than	NN	O	O
that	NN	O	O
seen	NN	O	O
in	NN	O	O
XSCID	NN	O	O
.	NN	O	O

-DOCSTART-	O

Posttranscriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
macrophage	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
antioxidants	NN	O	O
.	NN	O	O

Tissue	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
TF	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
by	NN	O	O
cells	NN	O	O
of	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
represents	NN	O	O
an	NN	O	O
important	NN	O	O
mechanism	NN	O	O
underlying	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
fibrin	NN	O	B-protein
deposition	NN	O	O
at	NN	O	O
sites	NN	O	O
of	NN	O	O
extravascular	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Recent	NN	O	O
evidence	NN	O	O
suggests	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
oxidant	NN	O	O
stress	NN	O	O
in	NN	O	O
the	NN	O	O
signalling	NN	O	O
pathway	NN	O	O
of	NN	O	O
various	NN	O	O
cell	NN	O	O
types	NN	O	O
by	NN	O	O
virtue	NN	O	O
of	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
various	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
antioxidant	NN	O	O
treatment	NN	O	O
on	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-induced	NN	O	O
TF	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
peritoneal	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Both	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
,	NN	O	O
an	NN	O	O
oxidant	NN	O	O
scavenger	NN	O	O
,	NN	O	O
and	NN	O	O
N-acetyl-cysteine	NN	O	O
,	NN	O	O
a	NN	O	O
precursor	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	O
antioxidant	NN	O	O
glutathione	NN	O	O
,	NN	O	O
inhibited	NN	O	O
stimulation	NN	O	O
of	NN	O	O
macrophage	NN	O	O
procoagulant	NN	O	O
activity	NN	O	O
by	NN	O	O
LPS	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
neither	NN	O	O
of	NN	O	O
these	NN	O	O
agents	NN	O	O
reduced	NN	O	O
LPS-stimulated	NN	O	O
TF	NN	O	B-protein
mRNA	NN	O	O
accumulation	NN	O	O
,	NN	O	O
thereby	NN	O	O
suggesting	NN	O	O
a	NN	O	O
posttranscriptional	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
effect	NN	O	O
.	NN	O	O

Immunofluorescence	NN	O	O
studies	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
using	NN	O	O
polyclonal	NN	O	O
anti-TF	NN	O	B-protein
antibody	NN	O	I-protein
showed	NN	O	O
that	NN	O	O
N-acetyl-cysteine	NN	O	O
treatment	NN	O	O
prevented	NN	O	O
the	NN	O	O
characteristic	NN	O	O
plasmalemmal	NN	O	O
localization	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
antigen	NN	O	I-protein
that	NN	O	O
occurs	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
N-acetyl-cysteine	NN	O	O
reduced	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
47-kD	NN	O	B-protein
mature	NN	O	I-protein
glycoprotein	NN	O	I-protein
in	NN	O	O
LPS-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
finding	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
the	NN	O	O
immunofluorescence	NN	O	O
studies	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
these	NN	O	O
conditions	NN	O	O
did	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
an	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
less	NN	O	O
mature	NN	O	O
forms	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
.	NN	O	O

When	NN	O	O
considered	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
antioxidants	NN	O	O
exert	NN	O	O
their	NN	O	O
effects	NN	O	O
by	NN	O	O
impairing	NN	O	O
translation	NN	O	O
and/or	NN	O	O
by	NN	O	O
causing	NN	O	O
degradation	NN	O	O
of	NN	O	O
newly	NN	O	O
translated	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
antioxidants	NN	O	O
on	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
species	NN	O	O
specific	NN	O	O
,	NN	O	O
with	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
LPS-induced	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor	NN	O	O
in	NN	O	O
murine	NN	O	O
cells	NN	O	O
,	NN	O	O
but	NN	O	O
with	NN	O	O
inhibition	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
posttranscriptional	NN	O	O
effect	NN	O	O
of	NN	O	O
antioxidants	NN	O	O
on	NN	O	O
TF	NN	O	B-protein
expression	NN	O	O
data	NN	O	O
suggests	NN	O	O
a	NN	O	O
novel	NN	O	O
mechanism	NN	O	O
whereby	NN	O	O
these	NN	O	O
agents	NN	O	O
might	NN	O	O
modulate	NN	O	O
monocyte	NN	O	B-cell_type
/macrophage	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
CD48	NN	O	B-DNA
gene	NN	O	I-DNA
demonstrates	NN	O	O
a	NN	O	O
positive	NN	O	O
element	NN	O	O
that	NN	O	O
is	NN	O	O
specific	NN	O	O
to	NN	O	O
Epstein-Barr	NN	O	B-cell_type
virus-immortalized	NN	O	I-cell_type
B-cell	NN	O	I-cell_type
lines	NN	O	I-cell_type
and	NN	O	O
contains	NN	O	O
an	NN	O	O
essential	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
infection	NN	O	O
of	NN	O	O
mature	NN	O	O
,	NN	O	O
resting	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
drives	NN	O	O
them	NN	O	O
to	NN	O	O
become	NN	O	O
lymphoblasts	NN	O	O
expressing	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
cell	NN	O	O
surface	NN	O	O
molecules	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
CD48	NN	O	B-protein
,	NN	O	O
characteristically	NN	O	O
expressed	NN	O	O
on	NN	O	O
normal	NN	O	B-cell_type
activated	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
on	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
an	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
CD48	NN	O	B-DNA
gene	NN	O	I-DNA
which	NN	O	O
reproducibly	NN	O	O
confers	NN	O	O
strong	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
only	NN	O	O
in	NN	O	O
EBV-positive	NN	O	B-cell_line
B-lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
element	NN	O	O
is	NN	O	O
not	NN	O	O
activated	NN	O	O
upon	NN	O	O
infection	NN	O	O
of	NN	O	O
established	NN	O	O
EBV-negative	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
EBV	NN	O	O
fails	NN	O	O
to	NN	O	O
drive	NN	O	O
these	NN	O	O
cells	NN	O	O
to	NN	O	O
a	NN	O	O
fully	NN	O	O
lymphoblastoid	NN	O	B-cell_type
phenotype	NN	O	I-cell_type
.	NN	O	O

An	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
site	NN	O	O
is	NN	O	O
an	NN	O	O
essential	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
element	NN	O	O
but	NN	O	O
alone	NN	O	O
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
activity	NN	O	O
or	NN	O	O
the	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
element	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
detected	NN	O	O
a	NN	O	O
specific	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
complex	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
element	NN	O	O
and	NN	O	O
show	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
(	NN	O	O
p50	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
part	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
.	NN	O	O

The	NN	O	O
EBV-encoded	NN	O	B-protein
latent	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
transactivating	NN	O	O
the	NN	O	O
isolated	NN	O	O
CD48	NN	O	B-protein
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
the	NN	O	O
intact	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
-driven	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
must	NN	O	O
interact	NN	O	O
with	NN	O	O
other	NN	O	O
regulatory	NN	O	O
pathways	NN	O	O
to	NN	O	O
control	NN	O	O
expression	NN	O	O
of	NN	O	O
cellular	NN	O	O
genes	NN	O	O
as	NN	O	O
EBV	NN	O	O
drives	NN	O	O
resting	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

-DOCSTART-	O

Thapsigargin	NN	O	O
induces	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
alpha-chain	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
and	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
via	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
-independent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Thapsigargin	NN	O	O
(	NN	O	O
TG	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
Ca	NN	O	B-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-ATPase	NN	O	I-protein
,	NN	O	O
depletes	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
stores	NN	O	O
and	NN	O	O
induces	NN	O	O
a	NN	O	O
sustained	NN	O	O
Ca2+	NN	O	O
influx	NN	O	O
without	NN	O	O
altering	NN	O	O
phosphatidyl	NN	O	O
inositol	NN	O	O
levels	NN	O	O
.	NN	O	O

TG	NN	O	O
plus	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
but	NN	O	O
not	NN	O	O
TG	NN	O	O
alone	NN	O	O
induced	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
TG	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
TG	NN	O	O
induced	NN	O	O
increases	NN	O	O
in	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
protein	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
IL-2R	NN	O	B-RNA
alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
by	NN	O	O
TG	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Further	NN	O	O
,	NN	O	O
like	NN	O	O
PMA	NN	O	O
,	NN	O	O
TG	NN	O	O
markedly	NN	O	O
induced	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
TG	NN	O	O
and	NN	O	O
PMA	NN	O	O
exhibited	NN	O	O
a	NN	O	O
synergistic	NN	O	O
action	NN	O	O
on	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
TG	NN	O	O
and	NN	O	O
PMA	NN	O	O
induce	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
through	NN	O	O
distinct	NN	O	O
pathways	NN	O	O
.	NN	O	O

PMA	NN	O	B-protein
-but	NN	O	I-protein
not	NN	O	I-protein
TG-induced	NN	O	I-protein
IL-2R	NN	O	I-protein
alpha	NN	O	I-protein
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
PKC	NN	O	B-protein
inhibitor	NN	O	O
H7	NN	O	O
,	NN	O	O
whereas	NN	O	O
TG	NN	O	B-protein
-but	NN	O	I-protein
not	NN	O	I-protein
PMA-induced	NN	O	I-protein
IL-2R	NN	O	I-protein
alpha	NN	O	I-protein
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
cholera	NN	O	O
toxin	NN	O	O
,	NN	O	O
forskolin	NN	O	O
and	NN	O	O
1	NN	O	O
,	NN	O	O
9-dideoxy	NN	O	O
forskolin	NN	O	O
.	NN	O	O

In	NN	O	O
toto	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
TG	NN	O	O
induces	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
a	NN	O	O
PKC	NN	O	B-protein
-independent	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Physiological	NN	O	O
concentration	NN	O	O
of	NN	O	O
estradiol	NN	O	O
inhibits	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocyte	NN	O	I-cell_type
chemotaxis	NN	O	O
via	NN	O	O
a	NN	O	O
receptor	NN	O	O
mediated	NN	O	O
system	NN	O	O
.	NN	O	O

Estrogen	NN	O	O
exhibits	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
actions	NN	O	O
,	NN	O	O
including	NN	O	O
immuno-modulatory	NN	O	O
effects	NN	O	O
,	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
estrogen	NN	O	O
exerts	NN	O	O
its	NN	O	O
anti-inflammatory	NN	O	O
effect	NN	O	O
is	NN	O	O
not	NN	O	O
yet	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
possible	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
estradiol	NN	O	O
acting	NN	O	O
via	NN	O	O
the	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
PMNs	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
agent	NN	O	O
,	NN	O	O
17	NN	O	O
beta-estradiol	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
17	NN	O	O
alpha-estradiol	NN	O	O
,	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
PMNs	NN	O	B-cell_type
chemotaxis	NN	O	O
to	NN	O	O
FMLP	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
(	NN	O	O
control	NN	O	O
vs	NN	O	O
estrogen	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
-	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
M	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

Physiological	NN	O	O
concentrations	NN	O	O
of	NN	O	O
estradiol	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
the	NN	O	O
chemotaxis	NN	O	O
of	NN	O	O
PMNs	NN	O	B-cell_type
(	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
mol	NN	O	O
)	NN	O	O
.	NN	O	O

Pre-incubation	NN	O	O
with	NN	O	O
clomiphene	NN	O	O
or	NN	O	O
tamoxifen	NN	O	O
which	NN	O	O
are	NN	O	O
estrogen	NN	O	O
receptor	NN	O	O
antagonists	NN	O	O
,	NN	O	O
eliminated	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
17	NN	O	O
beta-estradiol	NN	O	O
on	NN	O	O
the	NN	O	O
chemotaxis	NN	O	O
of	NN	O	O
PMNs	NN	O	B-cell_type
,	NN	O	O
restoring	NN	O	O
it	NN	O	O
to	NN	O	O
the	NN	O	O
control	NN	O	O
level	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
17	NN	O	O
beta-estradiol	NN	O	O
suppressed	NN	O	O
the	NN	O	O
chemotaxis	NN	O	O
of	NN	O	O
PMNs	NN	O	B-cell_type
by	NN	O	O
a	NN	O	O
receptor-dependent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
estradiol	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
plasma	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
PMNs	NN	O	B-cell_type
were	NN	O	O
drawn	NN	O	O
,	NN	O	O
showed	NN	O	O
a	NN	O	O
close	NN	O	O
,	NN	O	O
inverse	NN	O	O
correlation	NN	O	O
with	NN	O	O
the	NN	O	O
PMNs	NN	O	B-cell_type
chemotaxis	NN	O	O
to	NN	O	O
FMLP	NN	O	O
(	NN	O	O
r	NN	O	O
=	NN	O	O
-0.821	NN	O	O
p	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
.	NN	O	O

Estrogen	NN	O	O
may	NN	O	O
modify	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
during	NN	O	O
the	NN	O	O
normal	NN	O	O
menstrual	NN	O	O
cycle	NN	O	O
,	NN	O	O
not	NN	O	O
only	NN	O	O
during	NN	O	O
pregnancy	NN	O	O
,	NN	O	O
and	NN	O	O
influence	NN	O	O
inflammation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
the	NN	O	O
TCL1	NN	O	B-DNA
gene	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_type
malignancies	NN	O	O
.	NN	O	O

The	NN	O	O
TCL1	NN	O	B-DNA
locus	NN	O	I-DNA
on	NN	O	O
chromosome	NN	O	B-DNA
14q32.1	NN	O	I-DNA
is	NN	O	O
frequently	NN	O	O
involved	NN	O	O
in	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
and	NN	O	O
inversions	NN	O	O
with	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	B-DNA
receptor	NN	O	I-DNA
loci	NN	O	I-DNA
in	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemias	NN	O	O
and	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

The	NN	O	O
chromosome	NN	O	B-DNA
14	NN	O	I-DNA
region	NN	O	I-DNA
translocated	NN	O	O
or	NN	O	O
rearranged	NN	O	O
involves	NN	O	O
approximately	NN	O	O
350	NN	O	O
kb	NN	O	O
of	NN	O	O
DNA	NN	O	O
at	NN	O	O
chromosome	NN	O	O
band	NN	O	O
14q32.1	NN	O	O
.	NN	O	O

Within	NN	O	O
this	NN	O	O
region	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
a	NN	O	O
1.3-kb	NN	O	B-RNA
transcript	NN	O	I-RNA
,	NN	O	O
expressed	NN	O	O
only	NN	O	O
in	NN	O	O
restricted	NN	O	O
subsets	NN	O	O
of	NN	O	O
cells	NN	O	O
within	NN	O	O
the	NN	O	O
lymphoid	NN	O	B-cell_type
lineage	NN	O	I-cell_type
and	NN	O	O
expressed	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
carrying	NN	O	O
a	NN	O	O
t	NN	O	O
(	NN	O	O
14	NN	O	O
;	NN	O	O
14	NN	O	O
)	NN	O	O
(	NN	O	O
q11	NN	O	O
;	NN	O	O
q32	NN	O	O
)	NN	O	O
chromosome	NN	O	O
translocation	NN	O	O
or	NN	O	O
a	NN	O	O
inv	NN	O	O
(	NN	O	O
14	NN	O	O
)	NN	O	O
(	NN	O	O
q11	NN	O	O
;	NN	O	O
q32	NN	O	O
)	NN	O	O
chromosome	NN	O	O
inversion	NN	O	O
.	NN	O	O

The	NN	O	O
cognate	NN	O	B-DNA
cDNA	NN	O	I-DNA
sequence	NN	O	I-DNA
reveals	NN	O	O
an	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
of	NN	O	O
342	NN	O	O
nt	NN	O	O
encoding	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
of	NN	O	I-protein
14	NN	O	I-protein
kDa	NN	O	I-protein
.	NN	O	O

The	NN	O	O
TCL1	NN	O	B-DNA
gene	NN	O	I-DNA
sequence	NN	O	O
,	NN	O	O
which	NN	O	O
,	NN	O	O
to	NN	O	O
our	NN	O	O
knowledge	NN	O	O
,	NN	O	O
shows	NN	O	O
no	NN	O	O
sequence	NN	O	O
homology	NN	O	O
with	NN	O	O
other	NN	O	O
human	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
preferentially	NN	O	O
expressed	NN	O	O
early	NN	O	O
in	NN	O	O
T-	NN	O	B-cell_type
and	NN	O	I-cell_type
B-lymphocyte	NN	O	I-cell_type
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
region	NN	O	O
which	NN	O	O
directs	NN	O	O
the	NN	O	O
monocytic	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
colony-stimulating	NN	O	B-DNA
factor	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	I-DNA
macrophage	NN	O	I-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
)	NN	O	I-DNA
receptor	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
binds	NN	O	O
PEBP2/CBF	NN	O	B-protein
(	NN	O	O
AML1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
receptor	NN	O	O
for	NN	O	O
the	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
or	NN	O	O
colony-stimulating	NN	O	B-protein
factor	NN	O	I-protein
1	NN	O	I-protein
[	NN	O	O
CSF-1	NN	O	B-protein
]	NN	O	O
)	NN	O	O
is	NN	O	O
expressed	NN	O	O
from	NN	O	O
different	NN	O	O
promoters	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
placental	NN	O	B-cell_type
trophoblasts	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
monocyte-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CSF-1	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
regulated	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
by	NN	O	O
a	NN	O	O
tissue-specific	NN	O	B-DNA
promoter	NN	O	I-DNA
whose	NN	O	O
activity	NN	O	O
is	NN	O	O
stimulated	NN	O	O
by	NN	O	O
the	NN	O	O
monocyte/B-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
PU.1	NN	O	B-protein
(	NN	O	O
D.-E.Zhang	NN	O	O
,	NN	O	O
C.J.Hetherington	NN	O	O
,	NN	O	O
H.-M.Chen	NN	O	O
,	NN	O	O
and	NN	O	O
D.G.Tenen	NN	O	O
,	NN	O	O
Mol.Cell.	NN	O	O
Biol.14	NN	O	O
:	NN	O	O
373-381	NN	O	O
,	NN	O	O
1994	NN	O	O
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
the	NN	O	O
tissue	NN	O	O
specificity	NN	O	O
of	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
is	NN	O	O
also	NN	O	O
mediated	NN	O	O
by	NN	O	O
sequences	NN	O	O
in	NN	O	O
a	NN	O	O
region	NN	O	B-DNA
II	NN	O	I-DNA
(	NN	O	O
bp	NN	O	O
-88	NN	O	O
to	NN	O	O
-	NN	O	O
59	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
lies	NN	O	O
10	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
PU.1-binding	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
analyzed	NN	O	O
by	NN	O	O
DNase	NN	O	B-protein
footprinting	NN	O	O
,	NN	O	O
region	NN	O	B-DNA
II	NN	O	I-DNA
was	NN	O	O
protected	NN	O	O
preferentially	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
confirmed	NN	O	O
that	NN	O	O
region	NN	O	B-DNA
II	NN	O	I-DNA
interacts	NN	O	O
specifically	NN	O	O
with	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
from	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Two	NN	O	O
gel	NN	O	B-protein
shift	NN	O	I-protein
complexes	NN	O	I-protein
(	NN	O	O
Mono	NN	O	B-protein
A	NN	O	I-protein
and	NN	O	O
Mono	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
were	NN	O	O
formed	NN	O	O
with	NN	O	O
separate	NN	O	O
sequence	NN	O	O
elements	NN	O	O
within	NN	O	O
this	NN	O	O
region	NN	O	O
.	NN	O	O

Competition	NN	O	O
and	NN	O	O
supershift	NN	O	O
experiments	NN	O	O
indicate	NN	O	O
that	NN	O	O
Mono	NN	O	B-protein
B	NN	O	I-protein
contains	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
polyomavirus	NN	O	B-protein
enhancer-binding	NN	O	I-protein
protein	NN	O	I-protein
2/core-binding	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
PEBP2/CBF	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	I-protein
which	NN	O	O
includes	NN	O	O
the	NN	O	O
AML1	NN	O	O
gene	NN	O	O
product	NN	O	O
,	NN	O	O
while	NN	O	O
Mono	NN	O	B-protein
A	NN	O	I-protein
is	NN	O	O
a	NN	O	O
distinct	NN	O	O
complex	NN	O	O
preferentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Promoter	NN	O	B-DNA
constructs	NN	O	I-DNA
with	NN	O	O
mutations	NN	O	O
in	NN	O	O
these	NN	O	O
sequence	NN	O	O
elements	NN	O	O
were	NN	O	O
no	NN	O	O
longer	NN	O	O
expressed	NN	O	O
specifically	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
multimerized	NN	O	B-DNA
region	NN	O	I-DNA
II	NN	O	I-DNA
sequence	NN	O	I-DNA
elements	NN	O	I-DNA
enhanced	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
heterologous	NN	O	O
thymidine	NN	O	O
kinase	NN	O	O
promoter	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
tested	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
monocyte/B-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
PU.1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
Mono	NN	O	B-protein
A	NN	O	I-protein
and	NN	O	I-protein
Mono	NN	O	I-protein
B	NN	O	I-protein
protein	NN	O	I-protein
complexes	NN	O	I-protein
act	NN	O	O
in	NN	O	O
concert	NN	O	O
to	NN	O	O
regulate	NN	O	O
monocyte-specific	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
CSF-1	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
activation	NN	O	O
,	NN	O	O
prevents	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
human	NN	O	O
promyelocytic	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
thymocytes	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
potently	NN	O	O
blocks	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
agents	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
HL-60	NN	O	B-cell_line
,	NN	O	O
with	NN	O	O
10	NN	O	O
micrograms/mL	NN	O	O
etoposide	NN	O	O
or	NN	O	O
2	NN	O	O
microM	NN	O	O
1-beta-D-arabinofuranosylcytosine	NN	O	O
induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
within	NN	O	O
1	NN	O	O
hr	NN	O	O
and	NN	O	O
subsequently	NN	O	O
caused	NN	O	O
apoptosis	NN	O	O
within	NN	O	O
3-4	NN	O	O
hr	NN	O	O
.	NN	O	O

The	NN	O	O
simultaneous	NN	O	O
addition	NN	O	O
of	NN	O	O
50-500	NN	O	O
microM	NN	O	O
PDTC	NN	O	O
with	NN	O	O
these	NN	O	O
agents	NN	O	O
blocked	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
completely	NN	O	O
abrogated	NN	O	O
both	NN	O	O
morphologically	NN	O	O
apoptotic	NN	O	O
changes	NN	O	O
and	NN	O	O
internucleosomal	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
6	NN	O	O
hr	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
PDTC	NN	O	O
failed	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
endonuclease	NN	O	B-protein
activity	NN	O	O
contained	NN	O	O
in	NN	O	O
the	NN	O	O
whole	NN	O	O
cell	NN	O	O
lysates	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
PDTC	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
etoposide	NN	O	O
-and	NN	O	O
dexamethasone-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
at	NN	O	O
a	NN	O	O
concentration	NN	O	O
of	NN	O	O
1-10	NN	O	O
microM	NN	O	O
.	NN	O	O

Since	NN	O	O
PDTC	NN	O	O
has	NN	O	O
both	NN	O	O
antioxidant	NN	O	O
and	NN	O	O
metal-ion	NN	O	O
chelating	NN	O	O
activities	NN	O	O
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
N-acetyl-L-cysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
(	NN	O	O
antioxidant	NN	O	O
)	NN	O	O
or	NN	O	O
o-phenanthroline	NN	O	O
(	NN	O	O
OP	NN	O	O
)	NN	O	O
(	NN	O	O
metal-ion	NN	O	O
chelator	NN	O	O
)	NN	O	O
on	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
HL-60	NN	O	O
cells	NN	O	O
or	NN	O	O
thymocytes	NN	O	B-cell_type
with	NN	O	O
100-500	NN	O	O
microM	NN	O	O
OP	NN	O	O
for	NN	O	O
2	NN	O	O
hr	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
10-60	NN	O	O
mM	NN	O	O
NAC	NN	O	O
,	NN	O	O
suppressed	NN	O	O
subsequent	NN	O	O
occurrence	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
etoposide	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
process	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Overexpression	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C-zeta	NN	O	I-protein
stimulates	NN	O	O
leukemic	NN	O	B-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
.	NN	O	O

A	NN	O	O
function	NN	O	O
for	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C-zeta	NN	O	I-protein
(	NN	O	O
PKC-zeta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
nonresponsive	NN	O	O
atypical	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
subfamily	NN	O	I-protein
,	NN	O	O
in	NN	O	O
modulating	NN	O	O
differentiation	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_line
U937	NN	O	I-cell_line
cell	NN	O	I-cell_line
.	NN	O	O

Transfected	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
stably	NN	O	O
overexpressing	NN	O	O
PKC-zeta	NN	O	B-protein
displayed	NN	O	O
a	NN	O	O
longer	NN	O	O
doubling	NN	O	O
time	NN	O	O
,	NN	O	O
lower	NN	O	O
saturation	NN	O	O
density	NN	O	O
at	NN	O	O
confluency	NN	O	O
,	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
adherence	NN	O	O
to	NN	O	O
plastic	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
control	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

PKC-zeta	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressed	NN	O	O
a	NN	O	O
more	NN	O	O
differentiated	NN	O	O
phenotype	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
changes	NN	O	O
in	NN	O	O
morphology	NN	O	O
,	NN	O	O
surface	NN	O	O
antigen	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
lysosomal	NN	O	B-protein
enzyme	NN	O	I-protein
activities	NN	O	O
and	NN	O	O
were	NN	O	O
distinct	NN	O	O
from	NN	O	O
parental	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
stimulated	NN	O	O
to	NN	O	O
differentiate	NN	O	O
by	NN	O	O
exposure	NN	O	O
to	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
parental	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
PKC-zeta	NN	O	B-cell_line
cells	NN	O	I-cell_line
constitutively	NN	O	O
expressed	NN	O	O
mRNA	NN	O	B-RNA
transcripts	NN	O	I-RNA
for	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
a	NN	O	O
low	NN	O	O
mobility	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
PKC-zeta	NN	O	B-protein
overexpression	NN	O	O
stimulates	NN	O	O
a	NN	O	O
type	NN	O	O
of	NN	O	O
phenotypic	NN	O	O
differentiation	NN	O	O
that	NN	O	O
differs	NN	O	O
significantly	NN	O	O
from	NN	O	O
maturation	NN	O	O
occurring	NN	O	O
upon	NN	O	O
activation	NN	O	O
of	NN	O	O
other	NN	O	O
PKC	NN	O	B-protein
subfamilies	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
treatment	NN	O	O
.	NN	O	O

Increased	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
c-jun	NN	O	B-DNA
protooncogene	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
PKC-zeta	NN	O	B-cell_line
cells	NN	O	I-cell_line
provides	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
for	NN	O	O
explaining	NN	O	O
the	NN	O	O
altered	NN	O	O
differentiation	NN	O	O
status	NN	O	O
of	NN	O	O
this	NN	O	O
cell	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
family	NN	O	O
of	NN	O	O
serine	NN	O	B-protein
proteases	NN	O	I-protein
expressed	NN	O	O
exclusively	NN	O	O
in	NN	O	O
myelo-	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
specifically	NN	O	O
processes	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
subunit	NN	O	I-protein
p65	NN	O	I-protein
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
may	NN	O	O
impair	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
replication	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Two	NN	O	O
groups	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
obtained	NN	O	O
by	NN	O	O
limiting	NN	O	O
dilution	NN	O	O
cloning	NN	O	O
which	NN	O	O
differed	NN	O	O
strikingly	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
support	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
replication	NN	O	O
.	NN	O	O

``	NN	O	B-cell_line
Plus	NN	O	I-cell_line
''	NN	O	I-cell_line
clones	NN	O	I-cell_line
replicated	NN	O	O
the	NN	O	O
virus	NN	O	O
efficiently	NN	O	O
,	NN	O	O
whereas	NN	O	O
``	NN	O	B-cell_line
minus	NN	O	I-cell_line
''	NN	O	I-cell_line
clones	NN	O	I-cell_line
did	NN	O	O
not	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
these	NN	O	O
clones	NN	O	O
for	NN	O	O
differences	NN	O	O
in	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
activity	NN	O	O
which	NN	O	O
might	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
observed	NN	O	O
phenomenon	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
plus	NN	O	B-cell_line
clones	NN	O	I-cell_line
liberated	NN	O	O
the	NN	O	O
classical	NN	O	O
p50-p65	NN	O	B-protein
complex	NN	O	I-protein
from	NN	O	O
cytoplasmic	NN	O	O
pools	NN	O	O
,	NN	O	O
whereas	NN	O	O
minus	NN	O	B-cell_line
clones	NN	O	I-cell_line
produced	NN	O	O
an	NN	O	O
apparently	NN	O	O
novel	NN	O	O
,	NN	O	O
faster-migrating	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
as	NN	O	O
judged	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
surprising	NN	O	O
that	NN	O	O
the	NN	O	O
faster-migrating	NN	O	B-protein
complex	NN	O	I-protein
was	NN	O	O
composed	NN	O	O
also	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
subunit	NN	O	I-protein
was	NN	O	O
COOH-terminally	NN	O	O
truncated	NN	O	O
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
immunoprecipitation	NN	O	O
.	NN	O	O

The	NN	O	O
truncation	NN	O	O
resulted	NN	O	O
from	NN	O	O
limited	NN	O	O
proteolysis	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
during	NN	O	O
cellular	NN	O	O
extraction	NN	O	O
which	NN	O	O
released	NN	O	O
particular	NN	O	O
lysosomal	NN	O	B-protein
serine	NN	O	I-protein
proteases	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
elastase	NN	O	B-protein
,	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
,	NN	O	O
and	NN	O	O
proteinase	NN	O	B-protein
3	NN	O	I-protein
.	NN	O	O

These	NN	O	O
specific	NN	O	O
proteases	NN	O	O
are	NN	O	O
coordinately	NN	O	O
expressed	NN	O	O
and	NN	O	O
were	NN	O	O
present	NN	O	O
exclusively	NN	O	O
in	NN	O	O
the	NN	O	O
minus	NN	O	B-cell_line
U937	NN	O	I-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
plus	NN	O	B-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
these	NN	O	O
proteases	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
certain	NN	O	O
subclones	NN	O	O
of	NN	O	O
THP-1	NN	O	B-cell_line
and	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
primary	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
in	NN	O	O
each	NN	O	O
case	NN	O	O
correlating	NN	O	O
with	NN	O	O
the	NN	O	O
truncated	NN	O	O
from	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
in	NN	O	O
vitro	NN	O	O
cleavage	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
by	NN	O	O
purified	NN	O	O
elastase	NN	O	B-protein
and	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
possible	NN	O	O
that	NN	O	O
particular	NN	O	O
serine	NN	O	B-protein
proteases	NN	O	I-protein
may	NN	O	O
have	NN	O	O
inhibiting	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
replication	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
myelo-monocytic	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
data	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
special	NN	O	O
precautions	NN	O	O
must	NN	O	O
be	NN	O	O
taken	NN	O	O
when	NN	O	O
making	NN	O	O
extracts	NN	O	O
from	NN	O	O
myelo-monocytic	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
germline	NN	O	B-DNA
TaqI	NN	O	I-DNA
restriction	NN	O	I-DNA
fragment	NN	O	I-DNA
length	NN	O	I-DNA
polymorphism	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
progesterone	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
ovarian	NN	O	O
carcinoma	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Clinical	NN	O	O
outcome	NN	O	O
in	NN	O	O
ovarian	NN	O	O
carcinoma	NN	O	O
is	NN	O	O
predicted	NN	O	O
by	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
status	NN	O	O
,	NN	O	O
indicating	NN	O	O
an	NN	O	O
endocrine	NN	O	O
aspect	NN	O	O
to	NN	O	O
this	NN	O	O
disease	NN	O	O
.	NN	O	O

Peripheral	NN	O	B-DNA
leucocyte	NN	O	I-DNA
genomic	NN	O	I-DNA
DNAs	NN	O	I-DNA
were	NN	O	O
obtained	NN	O	O
from	NN	O	O
41	NN	O	O
patients	NN	O	O
with	NN	O	O
primary	NN	O	O
ovarian	NN	O	O
carcinoma	NN	O	O
and	NN	O	O
83	NN	O	O
controls	NN	O	O
from	NN	O	O
Ireland	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
from	NN	O	O
26	NN	O	O
primary	NN	O	O
ovarian	NN	O	O
carcinoma	NN	O	O
patients	NN	O	O
and	NN	O	O
101	NN	O	O
controls	NN	O	O
in	NN	O	O
Germany	NN	O	O
.	NN	O	O

Southern	NN	O	O
analysis	NN	O	O
using	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
progesterone	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
hPR	NN	O	I-DNA
)	NN	O	I-DNA
cDNA	NN	O	I-DNA
probe	NN	O	I-DNA
identified	NN	O	O
a	NN	O	O
germline	NN	O	B-DNA
TaqI	NN	O	I-DNA
restriction	NN	O	I-DNA
fragment	NN	O	I-DNA
length	NN	O	I-DNA
polymorphism	NN	O	I-DNA
(	NN	O	O
RFLP	NN	O	B-DNA
)	NN	O	O
defined	NN	O	O
by	NN	O	O
two	NN	O	O
alleles	NN	O	O
:	NN	O	O
T1	NN	O	B-DNA
,	NN	O	O
represented	NN	O	O
by	NN	O	O
a	NN	O	O
2.7	NN	O	B-DNA
kb	NN	O	I-DNA
fragment	NN	O	I-DNA
;	NN	O	O
and	NN	O	O
T2	NN	O	B-DNA
,	NN	O	O
represented	NN	O	O
by	NN	O	O
a	NN	O	O
1.9	NN	O	B-DNA
kb	NN	O	I-DNA
fragment	NN	O	I-DNA
and	NN	O	O
characterised	NN	O	O
by	NN	O	O
an	NN	O	O
additional	NN	O	O
TaqI	NN	O	B-DNA
restriction	NN	O	I-DNA
site	NN	O	I-DNA
with	NN	O	O
respect	NN	O	O
to	NN	O	O
T1	NN	O	B-DNA
.	NN	O	O

An	NN	O	O
over-representation	NN	O	O
of	NN	O	O
T2	NN	O	B-DNA
in	NN	O	O
ovarian	NN	O	O
cancer	NN	O	O
patients	NN	O	O
compared	NN	O	O
with	NN	O	O
controls	NN	O	O
in	NN	O	O
the	NN	O	O
pooled	NN	O	O
Irish/German	NN	O	O
population	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.025	NN	O	O
)	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

A	NN	O	O
difference	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.02	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
the	NN	O	O
RFLP	NN	O	B-DNA
genotypes	NN	O	O
between	NN	O	O
Irish	NN	O	O
and	NN	O	O
German	NN	O	O
control	NN	O	O
populations	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
.	NN	O	O

The	NN	O	O
allele	NN	O	O
distributions	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
shown	NN	O	O
to	NN	O	O
differ	NN	O	O
significantly	NN	O	O
from	NN	O	O
Hardy-Weinberg	NN	O	O
distribution	NN	O	O
in	NN	O	O
any	NN	O	O
subgroup	NN	O	O
.	NN	O	O

Using	NN	O	O
hPR	NN	O	B-DNA
cDNA	NN	O	I-DNA
region-specific	NN	O	I-DNA
probes	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
extra	NN	O	O
TaqI	NN	O	O
restriction	NN	O	O
site	NN	O	O
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
intron	NN	O	B-DNA
G	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
hPR	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

OBF-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
B	NN	O	I-protein
cell-specific	NN	O	I-protein
coactivator	NN	O	I-protein
that	NN	O	O
stimulates	NN	O	O
immunoglobulin	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
through	NN	O	O
association	NN	O	O
with	NN	O	O
octamer-binding	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Recent	NN	O	O
biochemical	NN	O	O
and	NN	O	O
genetic	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
octamer-binding	NN	O	B-protein
proteins	NN	O	I-protein
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
other	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
components	NN	O	I-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
lymphoid-restricted	NN	O	O
,	NN	O	O
octamer	NN	O	O
site-mediated	NN	O	O
immunoglobulin	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
genetic	NN	O	O
screen	NN	O	O
in	NN	O	O
yeast	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
isolated	NN	O	O
B	NN	O	B-DNA
cell-derived	NN	O	I-DNA
cDNAs	NN	O	I-DNA
encoding	NN	O	O
Oct-binding	NN	O	B-protein
factor	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
OBF-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
novel	NN	O	O
protein	NN	O	O
that	NN	O	O
specifically	NN	O	O
associates	NN	O	O
with	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
.	NN	O	O

Biochemical	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
OBF-1	NN	O	B-protein
has	NN	O	O
no	NN	O	O
intrinsic	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
and	NN	O	O
recognizes	NN	O	O
the	NN	O	O
POU	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
those	NN	O	O
of	NN	O	O
Oct-4	NN	O	B-protein
and	NN	O	O
Oct-6	NN	O	B-protein
.	NN	O	O

The	NN	O	O
OBF-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
highly	NN	O	O
cell-specific	NN	O	O
manner	NN	O	O
,	NN	O	O
being	NN	O	O
most	NN	O	O
abundant	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
essentially	NN	O	O
absent	NN	O	O
in	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
cells	NN	O	O
or	NN	O	O
tissues	NN	O	O
tested	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
OBF-1	NN	O	B-protein
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
selectively	NN	O	O
stimulates	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
natural	NN	O	O
immunoglobulin	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
an	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
OBF-1	NN	O	B-protein
has	NN	O	O
all	NN	O	O
the	NN	O	O
properties	NN	O	O
expected	NN	O	O
for	NN	O	O
a	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
transcriptional	NN	O	I-protein
coactivator	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Modulation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activity	NN	O	O
by	NN	O	O
intracellular	NN	O	O
glutathione	NN	O	O
levels	NN	O	O
and	NN	O	O
by	NN	O	O
variations	NN	O	O
of	NN	O	O
the	NN	O	O
extracellular	NN	O	O
cysteine	NN	O	O
supply	NN	O	O
.	NN	O	O

HIV-infected	NN	O	O
individuals	NN	O	O
and	NN	O	O
SIV-infected	NN	O	O
rhesus	NN	O	O
macaques	NN	O	O
have	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
average	NN	O	O
,	NN	O	O
decreased	NN	O	O
plasma	NN	O	O
cysteine	NN	O	O
and	NN	O	O
cystine	NN	O	O
concentrations	NN	O	O
and	NN	O	O
decreased	NN	O	O
intracellular	NN	O	O
glutathione	NN	O	O
levels	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
depletion	NN	O	O
of	NN	O	O
intracellular	NN	O	O
glutathione	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
Molt-4	NN	O	B-cell_line
)	NN	O	O
inhibits	NN	O	O
the	NN	O	O
activation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
incubation	NN	O	O
with	NN	O	O
increasing	NN	O	O
extracellular	NN	O	O
concentrations	NN	O	O
of	NN	O	O
cysteine	NN	O	O
inhibits	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
and	NN	O	O
transactivating	NN	O	O
activity	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Because	NN	O	O
inhibition	NN	O	O
of	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
increasing	NN	O	O
intracellular	NN	O	O
glutathione	NN	O	O
disulfide	NN	O	O
levels	NN	O	O
and	NN	O	O
GSSG	NN	O	B-protein
can	NN	O	O
be	NN	O	O
shown	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
directly	NN	O	O
in	NN	O	O
cell-free	NN	O	O
systems	NN	O	O
,	NN	O	O
our	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
GSSG	NN	O	B-protein
is	NN	O	O
a	NN	O	O
physiologically	NN	O	O
relevant	NN	O	O
inhibitor	NN	O	O
in	NN	O	O
intact	NN	O	B-cell_type
cells	NN	O	I-cell_type
also	NN	O	O
.	NN	O	O

NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
controls	NN	O	O
many	NN	O	O
immunologically	NN	O	B-DNA
important	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
so	NN	O	O
our	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
may	NN	O	O
be	NN	O	O
sensitive	NN	O	O
not	NN	O	O
only	NN	O	O
against	NN	O	O
a	NN	O	O
cysteine	NN	O	O
and	NN	O	O
glutathione	NN	O	O
deficiency	NN	O	O
but	NN	O	O
also	NN	O	O
against	NN	O	O
an	NN	O	O
excess	NN	O	O
of	NN	O	O
cysteine	NN	O	O
.	NN	O	O

-DOCSTART-	O

Two	NN	O	O
distinct	NN	O	O
signalling	NN	O	O
pathways	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
biphasic	NN	O	O
junB	NN	O	B-DNA
transcription	NN	O	O
induced	NN	O	O
by	NN	O	O
interleukin-6	NN	O	B-protein
in	NN	O	O
the	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridoma	NN	O	I-cell_line
7TD1	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
measured	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
junB	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
B	NN	O	B-cell_line
hybridoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
7TD1	NN	O	I-cell_line
,	NN	O	O
under	NN	O	O
interleukin-6	NN	O	B-protein
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
stimulation	NN	O	O
.	NN	O	O

IL-6	NN	O	B-protein
increases	NN	O	O
junB	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
a	NN	O	O
biphasic	NN	O	O
fashion	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
early-induced	NN	O	O
peak	NN	O	O
was	NN	O	O
transient	NN	O	O
and	NN	O	O
likely	NN	O	O
corresponds	NN	O	O
to	NN	O	O
the	NN	O	O
well	NN	O	O
documented	NN	O	O
typical	NN	O	O
junB	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
stimulated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
numerous	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
IL-6	NN	O	B-protein
.	NN	O	O

At	NN	O	O
variance	NN	O	O
,	NN	O	O
the	NN	O	O
second	NN	O	O
peak	NN	O	O
which	NN	O	O
has	NN	O	O
never	NN	O	O
been	NN	O	O
reported	NN	O	O
previously	NN	O	O
,	NN	O	O
lasted	NN	O	O
several	NN	O	O
hours	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
its	NN	O	O
effect	NN	O	O
on	NN	O	O
junB	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
IL-6	NN	O	B-protein
stimulated	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
biphasic	NN	O	O
fashion	NN	O	O
,	NN	O	O
the	NN	O	O
nuclear	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
JunB	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
IL-6	NN	O	B-protein
regulation	NN	O	O
occurred	NN	O	O
exclusively	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
bimodal	NN	O	O
increase	NN	O	O
of	NN	O	O
junB	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
JunB	NN	O	B-protein
protein	NN	O	I-protein
can	NN	O	O
be	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
a	NN	O	O
biphasic	NN	O	O
stimulation	NN	O	O
of	NN	O	O
junB	NN	O	B-DNA
transcription	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
point	NN	O	O
to	NN	O	O
two	NN	O	O
major	NN	O	O
differences	NN	O	O
between	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
early	NN	O	O
and	NN	O	O
the	NN	O	O
late	NN	O	O
IL-6	NN	O	B-protein
-induced	NN	O	O
junB	NN	O	B-DNA
transcription	NN	O	O
waves	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
cycloheximide	NN	O	O
strongly	NN	O	O
potentiated	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
second	NN	O	O
wave	NN	O	O
,	NN	O	O
whereas	NN	O	O
it	NN	O	O
failed	NN	O	O
to	NN	O	O
affect	NN	O	O
the	NN	O	O
early-induced	NN	O	O
burst	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
tyrphostin	NN	O	O
,	NN	O	O
a	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
impaired	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
second	NN	O	O
junB	NN	O	B-RNA
mRNA	NN	O	I-RNA
peak	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
genistein	NN	O	O
,	NN	O	O
another	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
totally	NN	O	O
abolished	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
second	NN	O	O
peak	NN	O	O
of	NN	O	O
junB	NN	O	B-RNA
mRNA	NN	O	I-RNA
whereas	NN	O	O
it	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
peak	NN	O	O
.	NN	O	O

Altogether	NN	O	O
these	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
7TD1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
IL-6	NN	O	B-protein
controls	NN	O	O
junB	NN	O	B-DNA
transcription	NN	O	O
in	NN	O	O
a	NN	O	O
biphasic	NN	O	O
fashion	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
two	NN	O	O
separate	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
newly	NN	O	O
established	NN	O	O
megakaryoblastic/erythroid	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
that	NN	O	O
differentiates	NN	O	O
to	NN	O	O
red	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
erythropoietin	NN	O	B-protein
and	NN	O	O
produces	NN	O	O
platelet-like	NN	O	B-cell_type
particles	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
August	NN	O	O
,	NN	O	O
1992	NN	O	O
,	NN	O	O
we	NN	O	O
established	NN	O	O
a	NN	O	O
leukemic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
NS-Meg	NN	O	B-cell_line
)	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
in	NN	O	O
megakaryoblastic	NN	O	O
transformation	NN	O	O
of	NN	O	O
Philadelphia	NN	O	O
chromosome-positive	NN	O	O
chronic	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
.	NN	O	O

The	NN	O	O
NS-Meg	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
positive	NN	O	O
for	NN	O	O
alpha-naphthyl	NN	O	B-protein
acetate	NN	O	I-protein
esterase	NN	O	I-protein
and	NN	O	O
periodic	NN	O	O
acid-Schiff	NN	O	O
(	NN	O	O
PAS	NN	O	O
)	NN	O	O
staining	NN	O	O
and	NN	O	O
for	NN	O	O
surface	NN	O	O
CD4	NN	O	B-protein
,	NN	O	O
CD7	NN	O	B-protein
,	NN	O	O
CD13	NN	O	B-protein
,	NN	O	O
CD34	NN	O	B-protein
,	NN	O	O
CD41a	NN	O	B-protein
,	NN	O	O
and	NN	O	O
glycophorin	NN	O	B-protein
A	NN	O	I-protein
antigens	NN	O	I-protein
.	NN	O	O

Ultrastructurally	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
had	NN	O	O
alpha-granules	NN	O	O
,	NN	O	O
demarcation	NN	O	O
membranes	NN	O	O
,	NN	O	O
and	NN	O	O
platelet	NN	O	B-protein
peroxidase	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

The	NN	O	O
NS-Meg	NN	O	B-cell_line
cells	NN	O	I-cell_line
spontaneously	NN	O	O
produced	NN	O	O
platelet-like	NN	O	B-cell_type
particles	NN	O	I-cell_type
which	NN	O	O
contained	NN	O	O
alpha-granules	NN	O	O
,	NN	O	O
mitochondria	NN	O	O
and	NN	O	O
dense	NN	O	O
bodies	NN	O	O
,	NN	O	O
strongly	NN	O	O
suggesting	NN	O	O
platelet	NN	O	O
production	NN	O	O
.	NN	O	O

Erythropoietin	NN	O	B-protein
(	NN	O	O
Epo	NN	O	B-protein
)	NN	O	O
,	NN	O	O
granulocyte/macrophage	NN	O	B-protein
colony	NN	O	I-protein
stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
interleukin	NN	O	B-protein
3	NN	O	I-protein
(	NN	O	O
IL-3	NN	O	B-protein
)	NN	O	O
promoted	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
NS-Meg	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Phorbol-12-myristate-13-acetate	NN	O	O
increased	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
CD41a	NN	O	B-protein
and	NN	O	O
CD61	NN	O	O
antigens	NN	O	O
.	NN	O	O

Ten-day	NN	O	O
exposure	NN	O	O
to	NN	O	O
Epo	NN	O	B-protein
induced	NN	O	O
mature	NN	O	B-cell_type
erythroblasts	NN	O	I-cell_type
and	NN	O	O
red	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
benzidine-positive	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
positive	NN	O	O
for	NN	O	O
hemoglobin	NN	O	B-protein
F	NN	O	O
staining	NN	O	O
.	NN	O	O

Untreated	NN	O	O
NS-Meg	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressed	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
the	NN	O	O
Epo	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
EpoR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
for	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
for	NN	O	O
alpha	NN	O	B-DNA
1	NN	O	I-DNA
,	NN	O	I-DNA
alpha	NN	O	I-DNA
2	NN	O	I-DNA
and	NN	O	I-DNA
gamma	NN	O	I-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
NS-Meg	NN	O	B-cell_line
cells	NN	O	I-cell_line
undergo	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
of	NN	O	O
both	NN	O	O
megakaryocytic	NN	O	B-cell_type
and	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
should	NN	O	O
be	NN	O	O
a	NN	O	O
very	NN	O	O
useful	NN	O	O
tool	NN	O	O
for	NN	O	O
the	NN	O	O
investigation	NN	O	O
of	NN	O	O
both	NN	O	O
megakaryocytic	NN	O	O
and	NN	O	O
erythroid	NN	O	O
maturation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
regulation	NN	O	O
of	NN	O	O
proto-oncogenes	NN	O	B-DNA
c-jun	NN	O	B-DNA
and	NN	O	O
c-fos	NN	O	B-DNA
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
activated	NN	O	O
through	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

The	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
molecule	NN	O	I-protein
CD28	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
ligands	NN	O	O
on	NN	O	O
accessory	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
APCs	NN	O	B-cell_type
,	NN	O	O
playing	NN	O	O
an	NN	O	O
important	NN	O	O
costimulatory	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
Ags	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
knowledge	NN	O	O
of	NN	O	O
the	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
coupled	NN	O	O
to	NN	O	O
this	NN	O	O
receptor	NN	O	O
is	NN	O	O
incomplete	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
phospholipase	NN	O	B-protein
C	NN	O	I-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
ligation	NN	O	O
of	NN	O	O
this	NN	O	O
receptor	NN	O	O
also	NN	O	O
seems	NN	O	O
to	NN	O	O
activate	NN	O	O
a	NN	O	O
calcium-independent	NN	O	O
,	NN	O	O
CD28	NN	O	B-protein
-specific	NN	O	O
pathway	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
(	NN	O	O
but	NN	O	O
not	NN	O	O
CD2	NN	O	B-protein
,	NN	O	O
CD5	NN	O	B-protein
,	NN	O	O
LFA-1	NN	O	B-protein
,	NN	O	O
or	NN	O	O
CD7	NN	O	B-protein
)	NN	O	O
leads	NN	O	O
to	NN	O	O
an	NN	O	O
elevation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
with	NN	O	O
only	NN	O	O
minimal	NN	O	O
activation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
.	NN	O	O

CD28	NN	O	B-protein
-dependent	NN	O	O
induction	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
requires	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
but	NN	O	O
does	NN	O	O
not	NN	O	O
depend	NN	O	O
on	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
phorbol	NN	O	B-protein
ester-responsive	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
or	NN	O	O
elevation	NN	O	O
of	NN	O	O
cytosolic	NN	O	O
calcium	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
CD28	NN	O	B-protein
-dependent	NN	O	O
elevation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
.	NN	O	O

A	NN	O	O
mechanism	NN	O	O
is	NN	O	O
suggested	NN	O	O
whereby	NN	O	O
expression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
junB	NN	O	B-DNA
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
fos	NN	O	B-DNA
family	NN	O	I-DNA
,	NN	O	O
can	NN	O	O
prevent	NN	O	O
inappropriate	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
caused	NN	O	O
by	NN	O	O
ligation	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
specific	NN	O	O
antigenic	NN	O	O
stimulus	NN	O	O
.	NN	O	O

-DOCSTART-	O

Enhanced	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
unique	NN	O	O
phenotype	NN	O	O
of	NN	O	O
simian	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
variant	NN	O	O
SIVsmmPBj14	NN	O	O
.	NN	O	O

Infection	NN	O	O
with	NN	O	O
a	NN	O	O
variant	NN	O	O
of	NN	O	O
simian	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
,	NN	O	O
SIVsmmPBj14	NN	O	O
,	NN	O	O
leads	NN	O	O
to	NN	O	O
severe	NN	O	O
acute	NN	O	O
disease	NN	O	O
in	NN	O	O
macaques	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
was	NN	O	O
designed	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
functional	NN	O	O
significance	NN	O	O
of	NN	O	O
previously	NN	O	O
described	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
to	NN	O	O
elucidate	NN	O	O
their	NN	O	O
contribution	NN	O	O
to	NN	O	O
the	NN	O	O
unique	NN	O	O
phenotype	NN	O	O
of	NN	O	O
SIVsmmPBj14	NN	O	O
.	NN	O	O

LTR	NN	O	B-DNA
-directed	NN	O	O
transcription	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
using	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
that	NN	O	O
were	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
into	NN	O	O
cultured	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
the	NN	O	O
basal	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
from	NN	O	O
SIVsmmPBj14	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
2-	NN	O	O
to	NN	O	O
4.5-fold	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
an	NN	O	O
LTR	NN	O	B-DNA
from	NN	O	O
a	NN	O	O
non-acutely	NN	O	O
pathogenic	NN	O	O
strain	NN	O	O
.	NN	O	O

These	NN	O	O
LTRs	NN	O	B-DNA
differ	NN	O	O
by	NN	O	O
five	NN	O	O
point	NN	O	O
mutations	NN	O	O
and	NN	O	O
a	NN	O	O
22-bp	NN	O	B-DNA
duplication	NN	O	I-DNA
in	NN	O	O
SIVsmmPBj14	NN	O	O
,	NN	O	O
which	NN	O	O
includes	NN	O	O
a	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
(	NN	O	I-DNA
NF	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Transcriptional	NN	O	O
differences	NN	O	O
between	NN	O	O
these	NN	O	O
LTRs	NN	O	B-DNA
were	NN	O	O
further	NN	O	O
enhanced	NN	O	O
by	NN	O	O
two-	NN	O	O
to	NN	O	O
threefold	NN	O	O
upon	NN	O	O
treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
or	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
or	NN	O	O
by	NN	O	O
cotransfection	NN	O	O
with	NN	O	O
plasmids	NN	O	B-DNA
expressing	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
subunits	NN	O	O
.	NN	O	O

Mutagenesis	NN	O	O
studies	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	O
construct	NN	O	O
containing	NN	O	O
an	NN	O	O
enhancerless	NN	O	O
promoter	NN	O	O
,	NN	O	O
indicate	NN	O	O
that	NN	O	O
these	NN	O	O
transcriptional	NN	O	O
effects	NN	O	O
are	NN	O	O
due	NN	O	O
principally	NN	O	O
to	NN	O	O
the	NN	O	O
22-bp	NN	O	B-DNA
sequence	NN	O	I-DNA
duplication	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
contained	NN	O	O
within	NN	O	O
it	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
infectious	NN	O	O
virus	NN	O	O
stocks	NN	O	O
that	NN	O	O
were	NN	O	O
isogenic	NN	O	O
except	NN	O	O
for	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
were	NN	O	O
generated	NN	O	O
.	NN	O	O

The	NN	O	O
LTR	NN	O	B-DNA
from	NN	O	O
SIVsmmPBj14	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
confer	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
virus	NN	O	O
replication	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Inclusion	NN	O	O
of	NN	O	O
this	NN	O	O
LTR	NN	O	B-DNA
in	NN	O	O
recombinant	NN	O	O
SIVs	NN	O	O
also	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
two-	NN	O	O
to	NN	O	O
threefold	NN	O	O
rise	NN	O	O
in	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
that	NN	O	O
was	NN	O	O
induced	NN	O	O
in	NN	O	O
quiescent	NN	O	B-cell_line
simian	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
blood	NN	O	I-cell_line
mononuclear	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
studies	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
LTR	NN	O	B-DNA
mutations	NN	O	O
assist	NN	O	O
SIVsmmPBj14	NN	O	O
in	NN	O	O
responding	NN	O	O
efficiently	NN	O	O
to	NN	O	O
cellular	NN	O	O
stimulation	NN	O	O
and	NN	O	O
allow	NN	O	O
it	NN	O	O
to	NN	O	O
replicate	NN	O	O
to	NN	O	O
high	NN	O	O
titers	NN	O	O
during	NN	O	O
the	NN	O	O
acute	NN	O	O
phase	NN	O	O
of	NN	O	O
viral	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte	NN	O	O
lineage	NN	O	O
.	NN	O	O

In	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
has	NN	O	O
been	NN	O	O
invoked	NN	O	O
as	NN	O	O
an	NN	O	O
important	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
of	NN	O	O
cell-surface	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
.	NN	O	O

In	NN	O	O
such	NN	O	O
cells	NN	O	O
,	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
can	NN	O	O
be	NN	O	O
detected	NN	O	O
without	NN	O	O
intentional	NN	O	O
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
our	NN	O	O
studies	NN	O	O
,	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
line	NN	O	I-cell_line
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
,	NN	O	O
cultured	NN	O	O
in	NN	O	O
medium	NN	O	O
containing	NN	O	O
fetal-calf	NN	O	O
serum	NN	O	O
and	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
also	NN	O	O
exhibit	NN	O	O
such	NN	O	O
'constitutive	NN	O	O
'	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
mobility-shift	NN	O	O
analysis	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
.	NN	O	O

This	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
was	NN	O	O
still	NN	O	O
present	NN	O	O
when	NN	O	O
contaminant	NN	O	O
LPS	NN	O	O
was	NN	O	O
removed	NN	O	O
by	NN	O	O
ultrafiltration	NN	O	O
and	NN	O	O
when	NN	O	O
serum	NN	O	O
was	NN	O	O
omitted	NN	O	O
.	NN	O	O

Protein-DNA	NN	O	O
complexes	NN	O	O
of	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
are	NN	O	O
similar	NN	O	O
in	NN	O	O
mobility	NN	O	O
to	NN	O	O
the	NN	O	O
LPS-induced	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
both	NN	O	O
are	NN	O	O
recognized	NN	O	O
by	NN	O	O
an	NN	O	O
antibody	NN	O	O
specific	NN	O	O
to	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
will	NN	O	O
only	NN	O	O
block	NN	O	O
LPS-induced	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
constitutive	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
LPS-free	NN	O	O
and	NN	O	O
serum-free	NN	O	O
conditions	NN	O	O
,	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
can	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
different	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
(	NN	O	O
HL60	NN	O	B-cell_line
,	NN	O	O
U937	NN	O	B-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
,	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
1	NN	O	I-cell_line
and	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
Molt	NN	O	B-cell_line
4	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
K562	NN	O	B-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

When	NN	O	O
ordered	NN	O	O
according	NN	O	O
to	NN	O	O
stage	NN	O	O
of	NN	O	O
maturation	NN	O	O
,	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
not	NN	O	O
increased	NN	O	O
in	NN	O	O
more	NN	O	O
mature	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
when	NN	O	O
inducing	NN	O	O
differentiation	NN	O	O
in	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
with	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
,	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
constitutive	NN	O	O
or	NN	O	O
inducible	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
can	NN	O	O
be	NN	O	O
detected	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
primary	NN	O	O
cells	NN	O	O
revealed	NN	O	O
substantial	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
blood	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
pleural	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
functionally	NN	O	O
active	NN	O	O
,	NN	O	O
since	NN	O	O
a	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
tumour	NN	O	B-RNA
necrosis	NN	O	I-RNA
factor	NN	O	I-RNA
(	NN	O	I-RNA
TNF	NN	O	I-RNA
)	NN	O	I-RNA
transcript	NN	O	I-RNA
is	NN	O	O
detectable	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
this	NN	O	O
level	NN	O	O
can	NN	O	O
be	NN	O	O
increased	NN	O	O
by	NN	O	O
blocking	NN	O	O
transcript	NN	O	O
degradation	NN	O	O
using	NN	O	O
cycloheximide	NN	O	O
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
is	NN	O	O
variable	NN	O	O
and	NN	O	O
is	NN	O	O
frequently	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
lower	NN	O	O
in	NN	O	O
the	NN	O	O
more	NN	O	O
mature	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
not	NN	O	O
maintained	NN	O	O
by	NN	O	O
autocrine	NN	O	O
action	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
TNF	NN	O	B-protein
,	NN	O	O
interleukin	NN	O	B-protein
6	NN	O	I-protein
,	NN	O	O
interleukin	NN	O	B-protein
10	NN	O	O
,	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
or	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
since	NN	O	O
neutralizing	NN	O	O
antibodies	NN	O	O
did	NN	O	O
not	NN	O	O
reduce	NN	O	O
constitutive	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
blockade	NN	O	O
of	NN	O	O
prostaglandin	NN	O	O
or	NN	O	O
leukotriene	NN	O	O
biosynthesis	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
400	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Steel	NN	O	B-protein
factor	NN	O	I-protein
affects	NN	O	O
SCL	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
normal	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Steel	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
controls	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
SCL	NN	O	B-protein
,	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
Tcl-5	NN	O	B-protein
or	NN	O	O
Tal-1	NN	O	B-protein
,	NN	O	O
is	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
erythropoiesis	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
SCL	NN	O	O
in	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
burst-forming	NN	O	I-cell_type
unit-erythroid	NN	O	I-cell_type
(	NN	O	O
BFU-E	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
Steel	NN	O	B-protein
factor	NN	O	I-protein
on	NN	O	O
SCL	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
proliferating	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

BFU-E-derived	NN	O	B-cell_line
colonies	NN	O	I-cell_line
increase	NN	O	O
progressively	NN	O	O
in	NN	O	O
size	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
cell	NN	O	O
number	NN	O	O
,	NN	O	O
from	NN	O	O
day	NN	O	O
7	NN	O	O
to	NN	O	O
day	NN	O	O
14	NN	O	O
of	NN	O	O
culture	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
greatest	NN	O	O
increase	NN	O	O
in	NN	O	O
colony	NN	O	O
size	NN	O	O
(	NN	O	O
10-fold	NN	O	O
expansion	NN	O	O
)	NN	O	O
occurring	NN	O	O
between	NN	O	O
day	NN	O	O
7	NN	O	O
and	NN	O	O
day	NN	O	O
10	NN	O	O
.	NN	O	O

SCL	NN	O	B-protein
protein	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
BFU-E-derived	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
highest	NN	O	O
in	NN	O	O
day	NN	O	O
7	NN	O	O
cells	NN	O	O
and	NN	O	O
decreased	NN	O	O
progressively	NN	O	O
from	NN	O	O
day	NN	O	O
7	NN	O	O
to	NN	O	O
day	NN	O	O
14	NN	O	O
of	NN	O	O
culture	NN	O	O
,	NN	O	O
suggesting	NN	O	O
an	NN	O	O
association	NN	O	O
of	NN	O	O
SCL	NN	O	B-protein
with	NN	O	O
erythroid	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
SCL	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
did	NN	O	O
not	NN	O	O
decrease	NN	O	O
significantly	NN	O	O
between	NN	O	O
day	NN	O	O
7	NN	O	O
and	NN	O	O
day	NN	O	O
14	NN	O	O
cells	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
posttranscriptional	NN	O	O
mechanisms	NN	O	O
are	NN	O	O
largely	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
decrease	NN	O	O
in	NN	O	O
SCL	NN	O	B-protein
protein	NN	O	I-protein
observed	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
SCL	NN	O	B-protein
in	NN	O	O
Steel	NN	O	B-protein
factor	NN	O	I-protein
-induced	NN	O	O
erythroid	NN	O	O
proliferation	NN	O	O
was	NN	O	O
then	NN	O	O
examined	NN	O	O
.	NN	O	O

In	NN	O	O
BFU-E-derived	NN	O	B-cell_line
colonies	NN	O	I-cell_line
cultured	NN	O	O
with	NN	O	O
Steel	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
colony	NN	O	O
size	NN	O	O
was	NN	O	O
significantly	NN	O	O
increased	NN	O	O
compared	NN	O	O
to	NN	O	O
control	NN	O	O
.	NN	O	O

In	NN	O	O
day	NN	O	O
7	NN	O	O
and	NN	O	O
day	NN	O	O
10	NN	O	O
erythroid	NN	O	B-cell_type
precursors	NN	O	I-cell_type
cultured	NN	O	O
with	NN	O	O
Steel	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
SCL	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
increased	NN	O	O
significantly	NN	O	O
compared	NN	O	O
to	NN	O	O
control	NN	O	O
.	NN	O	O

The	NN	O	O
increase	NN	O	O
in	NN	O	O
SCL	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
in	NN	O	O
early	NN	O	O
erythroid	NN	O	B-cell_type
precursors	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
Steel	NN	O	B-protein
factor	NN	O	I-protein
suggests	NN	O	O
one	NN	O	O
mechanism	NN	O	O
through	NN	O	O
which	NN	O	O
Steel	NN	O	B-protein
factor	NN	O	I-protein
may	NN	O	O
enhance	NN	O	O
normal	NN	O	O
erythroid	NN	O	O
proliferation	NN	O	O
.	NN	O	O

SCL	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
assessed	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
in	NN	O	O
day	NN	O	O
7	NN	O	O
cells	NN	O	O
did	NN	O	O
not	NN	O	O
increase	NN	O	O
significantly	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
Steel	NN	O	B-protein
factor	NN	O	I-protein
stimulation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
posttranscriptional	NN	O	O
mechanisms	NN	O	O
may	NN	O	O
also	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
SCL	NN	O	B-protein
protein	NN	O	I-protein
observed	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
Steel	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

C/EBP	NN	O	B-protein
beta	NN	O	I-protein
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Activated	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
contribute	NN	O	O
to	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
by	NN	O	O
the	NN	O	O
secretion	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
proteinases	NN	O	B-protein
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
is	NN	O	O
particularly	NN	O	O
important	NN	O	O
in	NN	O	O
this	NN	O	O
process	NN	O	O
because	NN	O	O
of	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
regulate	NN	O	O
other	NN	O	O
inflammatory	NN	O	O
mediators	NN	O	O
in	NN	O	O
an	NN	O	O
autocrine	NN	O	O
and	NN	O	O
paracrine	NN	O	O
fashion	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
is	NN	O	O
not	NN	O	O
known	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
data	NN	O	O
to	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
(	NN	O	O
NF-IL6	NN	O	B-protein
)	NN	O	O
.	NN	O	O

C/EBP	NN	O	B-protein
beta	NN	O	I-protein
activated	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
cotransfection	NN	O	O
assays	NN	O	O
and	NN	O	O
bound	NN	O	O
to	NN	O	O
it	NN	O	O
at	NN	O	O
a	NN	O	O
site	NN	O	O
which	NN	O	O
failed	NN	O	O
to	NN	O	O
bind	NN	O	O
the	NN	O	O
closely	NN	O	O
related	NN	O	O
protein	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
a	NN	O	O
dominant-negative	NN	O	O
version	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
blocked	NN	O	O
TNF	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
implicate	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
as	NN	O	O
an	NN	O	O
important	NN	O	O
regulator	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
by	NN	O	O
myelomonocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Calcium/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
II	NN	O	I-protein
downregulates	NN	O	O
both	NN	O	O
calcineurin	NN	O	O
and	NN	O	O
protein	NN	O	O
kinase	NN	O	O
C-mediated	NN	O	O
pathways	NN	O	O
for	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Engagement	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
for	NN	O	O
antigen	NN	O	O
activates	NN	O	O
phospholipase	NN	O	B-protein
C	NN	O	I-protein
resulting	NN	O	O
in	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
intracellular	NN	O	O
free	NN	O	O
calcium	NN	O	O
concentration	NN	O	O
(	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
)	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Increased	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
activates	NN	O	O
Ca2+/calmodulin-dependent	NN	O	B-protein
kinases	NN	O	I-protein
including	NN	O	O
the	NN	O	O
multifunctional	NN	O	O
Ca2+/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
II	NN	O	I-protein
(	NN	O	O
CaM-K	NN	O	B-protein
II	NN	O	I-protein
)	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
type	NN	O	B-protein
2B	NN	O	I-protein
protein	NN	O	I-protein
phosphatase	NN	O	I-protein
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
identified	NN	O	O
calcineurin	NN	O	B-protein
as	NN	O	O
a	NN	O	O
key	NN	O	O
enzyme	NN	O	O
for	NN	O	O
interleukin	NN	O	O
(	NN	O	O
IL	NN	O	O
)	NN	O	O
-2	NN	O	O
and	NN	O	O
IL-4	NN	O	O
promoter	NN	O	O
activation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
CaM-K	NN	O	B-protein
II	NN	O	I-protein
remains	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
mutants	NN	O	O
of	NN	O	O
these	NN	O	O
kinases	NN	O	B-protein
and	NN	O	O
phosphatases	NN	O	B-protein
(	NN	O	O
gamma	NN	O	B-protein
B*CaM-K	NN	O	I-protein
and	NN	O	O
delta	NN	O	B-protein
CaM-AI	NN	O	I-protein
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
to	NN	O	O
explore	NN	O	O
their	NN	O	O
relative	NN	O	O
role	NN	O	O
in	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
and	NN	O	O
their	NN	O	O
interactions	NN	O	O
with	NN	O	O
PKC	NN	O	B-protein
-dependent	NN	O	O
signaling	NN	O	O
systems	NN	O	O
.	NN	O	O

gamma	NN	O	B-protein
B*CaM-K	NN	O	I-protein
and	NN	O	O
delta	NN	O	B-protein
CaM-AI	NN	O	I-protein
,	NN	O	O
known	NN	O	O
to	NN	O	O
exhibit	NN	O	O
constitutive	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-independent	NN	O	O
activity	NN	O	O
,	NN	O	O
were	NN	O	O
cotransfected	NN	O	O
(	NN	O	O
alone	NN	O	O
or	NN	O	O
in	NN	O	O
combination	NN	O	O
)	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
a	NN	O	O
plasmid	NN	O	O
containing	NN	O	O
the	NN	O	O
intact	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
driving	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
gamma	NN	O	B-protein
B*CaM-K	NN	O	I-protein
with	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
construct	NN	O	I-DNA
downregulated	NN	O	O
its	NN	O	O
transcription	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
stimulation	NN	O	O
with	NN	O	O
ionomycin	NN	O	O
and	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
CaM-K	NN	O	B-protein
II	NN	O	I-protein
on	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
associated	NN	O	O
with	NN	O	O
decreased	NN	O	O
transcription	NN	O	O
of	NN	O	O
its	NN	O	O
AP-1	NN	O	O
and	NN	O	O
NF-AT	NN	O	O
transactivating	NN	O	O
pathways	NN	O	O
.	NN	O	O

Under	NN	O	O
the	NN	O	O
same	NN	O	O
conditions	NN	O	O
,	NN	O	O
delta	NN	O	B-protein
CaM-AI	NN	O	I-protein
superinduced	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
(	NN	O	O
approximately	NN	O	O
twofold	NN	O	O
increase	NN	O	O
)	NN	O	O
.	NN	O	O

When	NN	O	O
both	NN	O	O
mutants	NN	O	O
were	NN	O	O
used	NN	O	O
in	NN	O	O
combination	NN	O	O
,	NN	O	O
gamma	NN	O	B-protein
B*CaM-K	NN	O	I-protein
inhibited	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
delta	NN	O	B-protein
CaM-AI	NN	O	I-protein
.	NN	O	O

Similar	NN	O	O
results	NN	O	O
were	NN	O	O
obtained	NN	O	O
when	NN	O	O
a	NN	O	O
construct	NN	O	O
containing	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
also	NN	O	O
was	NN	O	O
used	NN	O	O
.	NN	O	O

gamma	NN	O	B-protein
B*CaM-K	NN	O	I-protein
also	NN	O	O
downregulated	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
transfection	NN	O	O
with	NN	O	O
a	NN	O	O
constitutively	NN	O	O
active	NN	O	O
mutant	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
or	NN	O	O
stimulation	NN	O	O
with	NN	O	O
PMA	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
CaM-K	NN	O	B-protein
II	NN	O	I-protein
may	NN	O	O
exert	NN	O	O
negative	NN	O	O
influences	NN	O	O
on	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
provide	NN	O	O
preliminary	NN	O	O
evidence	NN	O	O
for	NN	O	O
negative	NN	O	O
cross-talk	NN	O	O
with	NN	O	O
the	NN	O	O
calcineurin-	NN	O	O
and	NN	O	O
PKC-	NN	O	O
dependent	NN	O	O
signaling	NN	O	O
systems	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
and	NN	O	O
LFA-3	NN	O	B-protein
by	NN	O	O
Tax1	NN	O	B-protein
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
and	NN	O	O
mechanism	NN	O	O
of	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
or	NN	O	O
LFA-1	NN	O	B-protein
in	NN	O	O
adult-T-cell-leukemia	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
undertaken	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
HTLV-I	NN	O	B-protein
TaxI	NN	O	I-protein
in	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
ICAM-I	NN	O	B-protein
and	NN	O	O
LFA-3	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
transformed	NN	O	O
with	NN	O	O
HTLV-I	NN	O	O
and	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
ICAM-I	NN	O	B-protein
and	NN	O	O
LFA-I	NN	O	B-protein
in	NN	O	O
ATL-derived	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
TaxI	NN	O	B-protein
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
carrying	NN	O	O
the	NN	O	O
TaxI	NN	O	B-DNA
gene	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
metallothionein	NN	O	B-DNA
promoter	NN	O	I-DNA
led	NN	O	O
to	NN	O	O
increases	NN	O	O
in	NN	O	O
mRNA	NN	O	B-RNA
and	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
ICAM-I	NN	O	B-protein
.	NN	O	O

The	NN	O	O
response	NN	O	O
of	NN	O	O
LFA-3	NN	O	B-protein
to	NN	O	O
TaxI	NN	O	B-protein
induction	NN	O	O
was	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
relatively	NN	O	O
slow	NN	O	O
and	NN	O	O
weak	NN	O	O
,	NN	O	O
and	NN	O	O
might	NN	O	O
be	NN	O	O
indirect	NN	O	O
.	NN	O	O

Transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
ICAM-I	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
TaxI	NN	O	B-protein
was	NN	O	O
further	NN	O	O
shown	NN	O	O
by	NN	O	O
co-transfection	NN	O	O
of	NN	O	O
a	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
ICAM-I	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
plasmid	NN	O	B-DNA
expressing	NN	O	O
TaxI	NN	O	B-protein
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
ICAM-I	NN	O	B-protein
or	NN	O	O
LFA-I	NN	O	B-protein
in	NN	O	O
4	NN	O	B-cell_line
ATL	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
was	NN	O	O
next	NN	O	O
examined	NN	O	O
.	NN	O	O

ICAM-I	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
quite	NN	O	O
low	NN	O	O
in	NN	O	O
MT-I	NN	O	B-cell_line
,	NN	O	O
but	NN	O	O
no	NN	O	O
genomic	NN	O	O
changes	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

The	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
ICAM-I	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
inactive	NN	O	O
in	NN	O	O
MT-I	NN	O	B-cell_line
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
combined	NN	O	O
treatment	NN	O	O
of	NN	O	O
MT-I	NN	O	B-cell_line
with	NN	O	O
5-azacytidine	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
induced	NN	O	O
re-expression	NN	O	O
of	NN	O	O
ICAM-I	NN	O	B-protein
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
transcriptional	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
necessary	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
ICAM-I	NN	O	B-DNA
gene	NN	O	I-DNA
may	NN	O	O
be	NN	O	O
repressed	NN	O	O
in	NN	O	O
MT-I	NN	O	B-cell_line
through	NN	O	O
DNA	NN	O	O
methylation	NN	O	O
.	NN	O	O

Three	NN	O	O
other	NN	O	O
ATL	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
TL-OmI	NN	O	B-cell_line
,	NN	O	O
H582	NN	O	B-cell_line
,	NN	O	O
HuT102	NN	O	B-cell_line
)	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
little	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
the	NN	O	O
LFA-I	NN	O	B-protein
beta	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
CD18	NN	O	B-protein
)	NN	O	O
.	NN	O	O

H582	NN	O	B-cell_line
and	NN	O	O
HuT102	NN	O	B-cell_line
were	NN	O	O
also	NN	O	O
negative	NN	O	O
for	NN	O	O
the	NN	O	O
LFA-I	NN	O	B-RNA
alpha	NN	O	I-RNA
chain	NN	O	I-RNA
(	NN	O	I-RNA
CDIIa	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

No	NN	O	O
genomic	NN	O	O
changes	NN	O	O
were	NN	O	O
found	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
CD18	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
inactive	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
of	NN	O	O
them	NN	O	O
,	NN	O	O
again	NN	O	O
suggesting	NN	O	O
lack	NN	O	O
of	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
transcriptional	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
necessary	NN	O	O
for	NN	O	O
CD18	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Infection	NN	O	O
with	NN	O	O
Theileria	NN	O	O
annulata	NN	O	O
induces	NN	O	O
expression	NN	O	O
of	NN	O	O
matrix	NN	O	B-protein
metalloproteinase	NN	O	I-protein
9	NN	O	I-protein
and	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
in	NN	O	O
bovine	NN	O	B-cell_type
leucocytes	NN	O	I-cell_type
.	NN	O	O

Theileria	NN	O	O
annulata	NN	O	O
infects	NN	O	O
bovine	NN	O	B-cell_type
leucocytes	NN	O	I-cell_type
and	NN	O	O
results	NN	O	O
in	NN	O	O
their	NN	O	O
reversible	NN	O	O
transformation	NN	O	O
such	NN	O	O
that	NN	O	O
they	NN	O	O
become	NN	O	O
immortalised	NN	O	O
and	NN	O	O
metastatic	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
describes	NN	O	O
parasite-induced	NN	O	O
changes	NN	O	O
in	NN	O	O
host	NN	O	B-DNA
cell	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
which	NN	O	O
have	NN	O	O
a	NN	O	O
direct	NN	O	O
bearing	NN	O	O
on	NN	O	O
this	NN	O	O
transformation	NN	O	O
process	NN	O	O
.	NN	O	O

T.	NN	O	B-cell_type
annulata-infected	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
produce	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
novel	NN	O	O
metalloproteinase	NN	O	B-protein
activities	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
,	NN	O	O
previously	NN	O	O
called	NN	O	O
B1	NN	O	B-protein
,	NN	O	O
is	NN	O	O
a	NN	O	O
97-kDa	NN	O	B-protein
protein	NN	O	I-protein
which	NN	O	O
is	NN	O	O
secreted	NN	O	O
in	NN	O	O
large	NN	O	O
amounts	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
purified	NN	O	O
from	NN	O	O
protein-free	NN	O	O
,	NN	O	O
conditioned	NN	O	O
medium	NN	O	O
.	NN	O	O

An	NN	O	O
antiserum	NN	O	O
to	NN	O	O
this	NN	O	O
enzyme	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
isolate	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
predicted	NN	O	O
protein	NN	O	O
sequence	NN	O	O
of	NN	O	O
B1	NN	O	B-protein
is	NN	O	O
81	NN	O	O
%	NN	O	O
identical	NN	O	O
to	NN	O	O
human	NN	O	O
matrix	NN	O	B-protein
metalloproteinase	NN	O	I-protein
9	NN	O	I-protein
(	NN	O	O
MMP9	NN	O	B-protein
)	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
the	NN	O	O
bovine	NN	O	O
homologue	NN	O	O
of	NN	O	O
this	NN	O	O
enzyme	NN	O	O
.	NN	O	O

RNAase	NN	O	B-protein
protection	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
MMP9	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
unique	NN	O	O
to	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
increased	NN	O	O
MMP9	NN	O	B-protein
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
assayed	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
and	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
it	NN	O	O
was	NN	O	O
constitutively	NN	O	O
present	NN	O	O
in	NN	O	O
increased	NN	O	O
amounts	NN	O	O
in	NN	O	O
Theileria-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
we	NN	O	O
assayed	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
encoding	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
a	NN	O	O
common	NN	O	O
component	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
observed	NN	O	O
that	NN	O	O
it	NN	O	O
was	NN	O	O
indeed	NN	O	O
up-regulated	NN	O	O
in	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Since	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
and	NN	O	O
MMP9	NN	O	B-protein
can	NN	O	O
confer	NN	O	O
metastatic	NN	O	O
properties	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
are	NN	O	O
of	NN	O	O
considerable	NN	O	O
significance	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
transformed	NN	O	O
phenotype	NN	O	O
induced	NN	O	O
by	NN	O	O
Theileria	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

B-cell	NN	O	B-cell_type
proliferation	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
early	NN	O	O
G1-regulating	NN	O	B-protein
proteins	NN	O	I-protein
by	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
mutants	NN	O	O
conditional	NN	O	O
for	NN	O	O
EBNA2	NN	O	B-protein
.	NN	O	O

Infection	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
B-lymphocytes	NN	O	I-cell_type
by	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
leads	NN	O	O
to	NN	O	O
growth	NN	O	O
transformation	NN	O	O
of	NN	O	O
these	NN	O	O
B-cells	NN	O	B-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

EBV	NN	O	O
nuclear	NN	O	O
antigen	NN	O	O
2	NN	O	O
(	NN	O	O
EBNA2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
genes	NN	O	O
expressed	NN	O	O
after	NN	O	O
EBV	NN	O	O
infection	NN	O	O
of	NN	O	O
B-cells	NN	O	B-cell_type
,	NN	O	O
is	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
of	NN	O	O
viral	NN	O	B-DNA
and	NN	O	I-DNA
cellular	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
transforming	NN	O	O
potential	NN	O	O
of	NN	O	O
the	NN	O	O
virus	NN	O	O
.	NN	O	O

We	NN	O	O
generated	NN	O	O
conditional	NN	O	O
EBV	NN	O	O
mutants	NN	O	O
by	NN	O	O
expressing	NN	O	O
EBNA2	NN	O	B-protein
as	NN	O	O
chimeric	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
with	NN	O	O
the	NN	O	O
hormone	NN	O	O
binding	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
on	NN	O	O
the	NN	O	O
genetic	NN	O	O
background	NN	O	O
of	NN	O	O
the	NN	O	O
virus	NN	O	O
.	NN	O	O

Growth	NN	O	O
transformation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
normal	NN	O	I-cell_type
B-cells	NN	O	I-cell_type
by	NN	O	O
mutant	NN	O	O
virus	NN	O	O
resulted	NN	O	O
in	NN	O	O
estrogen-dependent	NN	O	B-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
expressing	NN	O	O
the	NN	O	O
chimeric	NN	O	O
EBNA2	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
estrogen	NN	O	O
about	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
enter	NN	O	O
a	NN	O	O
quiescent	NN	O	O
non-proliferative	NN	O	O
state	NN	O	O
whereas	NN	O	O
the	NN	O	O
others	NN	O	O
die	NN	O	O
by	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

EBNA2	NN	O	B-protein
is	NN	O	O
thus	NN	O	O
required	NN	O	O
not	NN	O	O
only	NN	O	O
for	NN	O	O
initiation	NN	O	O
but	NN	O	O
also	NN	O	O
for	NN	O	O
maintenance	NN	O	O
of	NN	O	O
transformation	NN	O	O
.	NN	O	O

Growth	NN	O	O
arrest	NN	O	O
occurred	NN	O	O
at	NN	O	O
G1	NN	O	O
and	NN	O	O
G2	NN	O	O
stages	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
functional	NN	O	O
EBNA2	NN	O	B-protein
is	NN	O	O
required	NN	O	O
at	NN	O	O
different	NN	O	O
restriction	NN	O	O
points	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

Growth	NN	O	O
arrest	NN	O	O
is	NN	O	O
reversible	NN	O	O
for	NN	O	O
G1/G0	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
indicated	NN	O	O
by	NN	O	O
the	NN	O	O
sequential	NN	O	O
accumulation	NN	O	O
and	NN	O	O
modification	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
cycle	NN	O	I-protein
regulating	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

EBV	NN	O	O
induces	NN	O	O
the	NN	O	O
same	NN	O	O
cell	NN	O	O
cycle	NN	O	O
regulating	NN	O	O
proteins	NN	O	O
as	NN	O	O
polyclonal	NN	O	O
stimuli	NN	O	O
in	NN	O	O
primary	NN	O	O
B-cells	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
EBV	NN	O	O
is	NN	O	O
using	NN	O	O
a	NN	O	O
common	NN	O	O
pathway	NN	O	O
for	NN	O	O
B-cell	NN	O	O
activation	NN	O	O
bypassing	NN	O	O
the	NN	O	O
requirement	NN	O	O
for	NN	O	O
antigen	NN	O	O
,	NN	O	O
T-cell	NN	O	O
signals	NN	O	O
and	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

IL-1	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
TCR	NN	O	B-protein
signals	NN	O	O
synergize	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-mediated	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-1R	NN	O	B-protein
)	NN	O	O
-	NN	O	O
and	NN	O	O
TCR	NN	O	B-protein
-initiated	NN	O	O
signals	NN	O	O
can	NN	O	O
interact	NN	O	O
synergistically	NN	O	O
to	NN	O	O
increase	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
several	NN	O	O
lymphokine	NN	O	B-DNA
and	NN	O	I-DNA
lymphokine	NN	O	I-DNA
receptor	NN	O	I-DNA
genes	NN	O	I-DNA
during	NN	O	O
the	NN	O	O
competence	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
activation	NN	O	O
program	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
helper	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
describe	NN	O	O
how	NN	O	O
signals	NN	O	O
initiated	NN	O	O
through	NN	O	O
the	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
IL-1R	NN	O	I-protein
interact	NN	O	O
with	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
to	NN	O	O
synergistically	NN	O	O
augment	NN	O	O
the	NN	O	O
transactivating	NN	O	O
properties	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
synergistic	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
initiated	NN	O	O
signals	NN	O	O
are	NN	O	O
mediated	NN	O	O
through	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
because	NN	O	O
they	NN	O	O
can	NN	O	O
be	NN	O	O
mimicked	NN	O	O
by	NN	O	O
the	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
,	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
calcium	NN	O	O
ionophores	NN	O	O
;	NN	O	O
and	NN	O	O
are	NN	O	O
staurosporine	NN	O	O
sensitive	NN	O	O
but	NN	O	O
cyclosporine	NN	O	O
resistant	NN	O	O
.	NN	O	O

Gel	NN	O	O
shift	NN	O	O
analyses	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
nuclear	NN	O	O
translocation	NN	O	O
is	NN	O	O
stimulated	NN	O	O
primarily	NN	O	O
by	NN	O	O
IL-1	NN	O	B-protein
rather	NN	O	O
than	NN	O	O
by	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
signals	NN	O	O
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
and	NN	O	O
phosphorylation	NN	O	O
analyses	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
synergistic	NN	O	O
effect	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
functional	NN	O	O
activity	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
MAD3	NN	O	B-protein
)	NN	O	O
-	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
dissociation	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

The	NN	O	O
IL-1	NN	O	B-protein
-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
is	NN	O	O
transient	NN	O	O
and	NN	O	O
can	NN	O	O
be	NN	O	O
prolonged	NN	O	O
either	NN	O	O
by	NN	O	O
an	NN	O	O
antigen	NN	O	O
receptor-initiated	NN	O	O
signal	NN	O	O
or	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
IL-1	NN	O	B-protein
induces	NN	O	O
both	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
responsible	NN	O	O
for	NN	O	O
terminating	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-DNA	NN	O	O
interaction	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Antigen	NN	O	O
receptor	NN	O	O
signals	NN	O	O
prolong	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-DNA	NN	O	O
interaction	NN	O	O
,	NN	O	O
probably	NN	O	O
by	NN	O	O
functionally	NN	O	O
antagonizing	NN	O	O
the	NN	O	O
IL-1	NN	O	B-protein
-induced	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
transient	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-DNA	NN	O	O
interaction	NN	O	O
and	NN	O	O
consequently	NN	O	O
synergistically	NN	O	O
enhance	NN	O	O
IL-1	NN	O	B-protein
-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
chicken	NN	O	O
anemia	NN	O	O
virus	NN	O	O
(	NN	O	O
CAV	NN	O	O
)	NN	O	O
containing	NN	O	O
a	NN	O	O
novel	NN	O	O
enhancer-like	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
single	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
cloned	NN	O	O
genome	NN	O	O
[	NN	O	O
Noteborn	NN	O	O
et	NN	O	O
al.	NN	O	O
,	NN	O	O
J.	NN	O	O
Virol.	NN	O	O
65	NN	O	O
(	NN	O	O
1991	NN	O	O
)	NN	O	O
3131-3139	NN	O	O
]	NN	O	O
of	NN	O	O
chicken	NN	O	O
anemia	NN	O	O
virus	NN	O	O
(	NN	O	O
CAV	NN	O	O
)	NN	O	O
in	NN	O	O
chicken	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
was	NN	O	O
analysed	NN	O	O
via	NN	O	O
CAT	NN	O	B-protein
assays	NN	O	O
.	NN	O	O

A	NN	O	O
unique	NN	O	O
region	NN	O	O
containing	NN	O	O
four	NN	O	O
or	NN	O	O
five	NN	O	O
near-perfect	NN	O	O
direct	NN	O	O
repeats	NN	O	O
(	NN	O	O
DR	NN	O	O
)	NN	O	O
of	NN	O	O
21	NN	O	O
bp	NN	O	O
with	NN	O	O
one	NN	O	O
12-bp	NN	O	B-DNA
insert	NN	O	I-DNA
was	NN	O	O
proven	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
main	NN	O	O
transcription-activation	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
with	NN	O	O
enhancer-like	NN	O	O
characteristics	NN	O	O
.	NN	O	O

PCR	NN	O	O
studies	NN	O	O
revealed	NN	O	O
that	NN	O	O
CAV	NN	O	O
isolates	NN	O	O
from	NN	O	O
across	NN	O	O
the	NN	O	O
world	NN	O	O
all	NN	O	O
contained	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Electrophoretic	NN	O	O
mobility-shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
showed	NN	O	O
that	NN	O	O
individual	NN	O	B-DNA
DR	NN	O	I-DNA
units	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
12-bp	NN	O	B-DNA
insert	NN	O	I-DNA
,	NN	O	O
can	NN	O	O
bind	NN	O	O
to	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
chicken	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

Competition	NN	O	O
assays	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
DR	NN	O	B-DNA
units	NN	O	I-DNA
bound	NN	O	O
to	NN	O	O
factors	NN	O	O
other	NN	O	O
than	NN	O	O
the	NN	O	O
12-bp	NN	O	B-DNA
insert	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
synthetic	NN	O	O
oligodeoxyribonucleotide	NN	O	O
containing	NN	O	O
an	NN	O	O
SP1-box	NN	O	B-DNA
(	NN	O	O
5'-GGGCGG	NN	O	O
)	NN	O	O
could	NN	O	O
compete	NN	O	O
with	NN	O	O
factors	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
12-bp	NN	O	B-DNA
insert	NN	O	I-DNA
.	NN	O	O

Purified	NN	O	O
human	NN	O	B-protein
SP1	NN	O	I-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
have	NN	O	O
very	NN	O	O
strong	NN	O	O
affinity	NN	O	O
for	NN	O	O
the	NN	O	O
12-bp	NN	O	B-DNA
insert	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
Myc-Max	NN	O	B-protein
heterodimer	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
activation-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
.	NN	O	O

T	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
respond	NN	O	O
to	NN	O	O
activation	NN	O	O
signals	NN	O	O
by	NN	O	O
undergoing	NN	O	O
apoptotic	NN	O	O
cell	NN	O	O
death	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
represent	NN	O	O
comparable	NN	O	O
events	NN	O	O
related	NN	O	O
to	NN	O	O
tolerance	NN	O	O
induction	NN	O	O
in	NN	O	O
immature	NN	O	B-cell_type
and	NN	O	I-cell_type
mature	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
using	NN	O	O
antisense	NN	O	O
oligonucleotides	NN	O	O
implicated	NN	O	O
the	NN	O	O
c-Myc	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
the	NN	O	O
phenomenon	NN	O	O
of	NN	O	O
activation-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

This	NN	O	O
role	NN	O	O
for	NN	O	O
c-Myc	NN	O	B-protein
in	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
now	NN	O	O
confirmed	NN	O	O
in	NN	O	O
studies	NN	O	O
using	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
form	NN	O	O
of	NN	O	O
its	NN	O	O
heterodimeric	NN	O	O
binding	NN	O	O
partner	NN	O	O
,	NN	O	O
Max	NN	O	B-protein
,	NN	O	O
which	NN	O	O
we	NN	O	O
show	NN	O	O
here	NN	O	O
inhibits	NN	O	O
activation-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
coexpression	NN	O	O
of	NN	O	O
a	NN	O	O
reciprocally	NN	O	B-protein
mutant	NN	O	I-protein
Myc	NN	O	I-protein
protein	NN	O	I-protein
capable	NN	O	O
of	NN	O	O
forming	NN	O	O
functional	NN	O	O
heterodimers	NN	O	B-protein
with	NN	O	O
the	NN	O	O
mutant	NN	O	O
Max	NN	O	B-protein
can	NN	O	O
compensate	NN	O	O
for	NN	O	O
the	NN	O	O
dominant	NN	O	O
negative	NN	O	O
activity	NN	O	O
and	NN	O	O
restore	NN	O	O
activation-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
imply	NN	O	O
that	NN	O	O
Myc	NN	O	B-protein
promotes	NN	O	O
activation-induced	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
obligatory	NN	O	O
heterodimerization	NN	O	O
with	NN	O	O
Max	NN	O	B-protein
,	NN	O	O
and	NN	O	O
therefore	NN	O	O
,	NN	O	O
by	NN	O	O
regulating	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
SM	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
protein	NN	O	O
products	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
BMLF1	NN	O	O
open	NN	O	O
reading	NN	O	O
frame	NN	O	O
have	NN	O	O
been	NN	O	O
characterized	NN	O	O
in	NN	O	O
the	NN	O	O
early	NN	O	O
productive	NN	O	O
cycle	NN	O	O
in	NN	O	O
B95-8	NN	O	B-cell_line
and	NN	O	O
Akata	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
SM	NN	O	B-protein
protein	NN	O	I-protein
derived	NN	O	O
from	NN	O	O
the	NN	O	O
spliced	NN	O	O
RNA	NN	O	O
joining	NN	O	O
BSLF2	NN	O	B-protein
to	NN	O	O
BMLF1	NN	O	B-protein
is	NN	O	O
much	NN	O	O
the	NN	O	O
most	NN	O	O
abundant	NN	O	O
protein	NN	O	O
.	NN	O	O

SM	NN	O	B-protein
is	NN	O	O
a	NN	O	O
phosphoprotein	NN	O	B-protein
in	NN	O	O
EBV	NN	O	O
-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
can	NN	O	O
be	NN	O	O
phosphorylated	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
casein	NN	O	B-protein
kinase	NN	O	I-protein
II	NN	O	I-protein
(	NN	O	O
CKII	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Computer	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
SM	NN	O	B-protein
protein	NN	O	I-protein
sequence	NN	O	O
showed	NN	O	O
a	NN	O	O
C	NN	O	B-protein
terminal	NN	O	I-protein
section	NN	O	I-protein
of	NN	O	O
SM	NN	O	B-protein
to	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
genome	NN	O	B-protein
positional	NN	O	I-protein
homologues	NN	O	I-protein
of	NN	O	O
four	NN	O	O
other	NN	O	O
herpesviruses	NN	O	O
and	NN	O	O
revealed	NN	O	O
consensus	NN	O	B-protein
CKII	NN	O	I-protein
sites	NN	O	I-protein
near	NN	O	O
the	NN	O	O
N	NN	O	B-protein
termini	NN	O	I-protein
of	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
SM	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
the	NN	O	O
herpes	NN	O	B-protein
simplex	NN	O	I-protein
virus	NN	O	I-protein
(	NN	O	I-protein
HSV	NN	O	I-protein
)	NN	O	I-protein
ICP27	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
the	NN	O	O
herpesvirus	NN	O	B-protein
saimiri	NN	O	I-protein
(	NN	O	I-protein
HVS	NN	O	I-protein
)	NN	O	I-protein
open	NN	O	I-protein
reading	NN	O	I-protein
frame	NN	O	I-protein
57	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
CKII	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
EBV	NN	O	O
SM	NN	O	O
greatly	NN	O	O
reduced	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
SM	NN	O	B-protein
by	NN	O	O
CKII	NN	O	B-protein
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
transactivation	NN	O	O
by	NN	O	O
BMLF1	NN	O	B-protein
proteins	NN	O	I-protein
has	NN	O	O
been	NN	O	O
controversial	NN	O	O
but	NN	O	O
SM	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
transactivate	NN	O	O
gene	NN	O	O
expression	NN	O	O
from	NN	O	O
a	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
by	NN	O	O
increasing	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	B-RNA
CAT	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
CKII	NN	O	B-protein
site	NN	O	O
in	NN	O	O
SM	NN	O	B-protein
made	NN	O	O
no	NN	O	O
difference	NN	O	O
to	NN	O	O
the	NN	O	O
transactivation	NN	O	O
in	NN	O	O
this	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assay	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inducible	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
c-fos	NN	O	O
serum	NN	O	O
response	NN	O	O
element	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
a	NN	O	O
phosphotyrosine-containing	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
proto-oncogene	NN	O	B-DNA
c-fos	NN	O	I-DNA
is	NN	O	O
an	NN	O	O
immediate-early	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
genes	NN	O	O
transcribed	NN	O	O
after	NN	O	O
stimulation	NN	O	O
of	NN	O	O
most	NN	O	O
cells	NN	O	O
with	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
ligands	NN	O	O
.	NN	O	O

Fos	NN	O	O
expression	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
pivotal	NN	O	O
event	NN	O	O
in	NN	O	O
converting	NN	O	O
ligand-receptor	NN	O	O
interactions	NN	O	O
at	NN	O	O
the	NN	O	O
membrane	NN	O	O
into	NN	O	O
functional	NN	O	O
modulation	NN	O	O
of	NN	O	O
cell	NN	O	O
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
serum	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
SRE	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
participates	NN	O	O
in	NN	O	O
induction	NN	O	O
of	NN	O	O
transcription	NN	O	O
by	NN	O	O
various	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
and	NN	O	O
subsequent	NN	O	O
squelching	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
an	NN	O	O
inducible	NN	O	O
protein	NN	O	B-protein
complex	NN	O	I-protein
(	NN	O	O
Band	NN	O	B-protein
A	NN	O	I-protein
)	NN	O	O
binds	NN	O	O
to	NN	O	O
SRE	NN	O	B-DNA
DNA	NN	O	I-DNA
within	NN	O	O
10	NN	O	O
min	NN	O	O
after	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
PBL-T	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
becomes	NN	O	O
nondetectable	NN	O	O
by	NN	O	O
60	NN	O	O
min	NN	O	O
.	NN	O	O

Band	NN	O	B-protein
A	NN	O	I-protein
contains	NN	O	O
the	NN	O	O
serum	NN	O	B-protein
response	NN	O	I-protein
factor	NN	O	I-protein
plus	NN	O	O
additional	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
protein	NN	O	O
that	NN	O	O
is	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
a	NN	O	O
tyrosine	NN	O	O
residue	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_line
PBL-T	NN	O	I-cell_line
suppresses	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
Band	NN	O	B-protein
A	NN	O	I-protein
to	NN	O	O
the	NN	O	O
SRE	NN	O	B-DNA
motif	NN	O	I-DNA
.	NN	O	O

Upon	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
,	NN	O	O
this	NN	O	O
protein	NN	O	O
no	NN	O	O
longer	NN	O	O
prevents	NN	O	O
binding	NN	O	O
of	NN	O	O
DNA	NN	O	O
by	NN	O	O
Band	NN	O	B-protein
A	NN	O	I-protein
,	NN	O	O
and	NN	O	O
suppression	NN	O	O
of	NN	O	O
binding	NN	O	O
is	NN	O	O
restored	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
.	NN	O	O

The	NN	O	O
phosphorylated	NN	O	O
tyrosine	NN	O	O
residue	NN	O	O
itself	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
protein-protein	NN	O	O
interaction	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
modification	NN	O	O
of	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
DRA	NN	O	I-DNA
promoter	NN	O	I-DNA
occupancy	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
AIR-1	NN	O	B-protein
trans-activator	NN	O	I-protein
.	NN	O	O

RJ	NN	O	B-cell_line
2.2.5	NN	O	I-cell_line
is	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
mutant	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
the	NN	O	O
Burkitt	NN	O	B-cell_line
lymphoma	NN	O	I-cell_line
Raji	NN	O	I-cell_line
cell	NN	O	I-cell_line
which	NN	O	O
is	NN	O	O
defective	NN	O	O
in	NN	O	O
the	NN	O	O
AIR-1	NN	O	O
locus	NN	O	O
function	NN	O	O
.	NN	O	O

This	NN	O	O
locus	NN	O	O
encodes	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
trans-activator	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
(	NN	O	I-DNA
MHC	NN	O	I-DNA
)	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
,	NN	O	O
by	NN	O	O
in	NN	O	O
vivo	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
footprinting	NN	O	O
,	NN	O	O
that	NN	O	O
the	NN	O	O
AIR-1	NN	O	B-DNA
locus	NN	O	I-DNA
defect	NN	O	I-DNA
correlates	NN	O	O
with	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
occupancy	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
reexpression	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
MHC	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
RJ	NN	O	B-cell_line
2.2.5	NN	O	I-cell_line
x	NN	O	I-cell_line
mouse	NN	O	I-cell_line
spleen	NN	O	I-cell_line
cell	NN	O	I-cell_line
hybrids	NN	O	I-cell_line
is	NN	O	O
associated	NN	O	O
with	NN	O	O
partial	NN	O	O
reversion	NN	O	O
of	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
occupancy	NN	O	O
to	NN	O	O
the	NN	O	O
Raji	NN	O	O
pattern	NN	O	O
.	NN	O	O

DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
occupancy	NN	O	O
in	NN	O	O
other	NN	O	O
class	NN	O	B-cell_line
II-negative	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
derived	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
bare	NN	O	O
lymphocyte	NN	O	O
syndrome	NN	O	O
,	NN	O	O
is	NN	O	O
drastically	NN	O	O
different	NN	O	O
from	NN	O	O
the	NN	O	O
one	NN	O	O
observed	NN	O	O
in	NN	O	O
RJ	NN	O	B-cell_line
2.2.5	NN	O	I-cell_line
and	NN	O	O
Raji	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
as	NN	O	O
an	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprinting	NN	O	O
agent	NN	O	O
reveals	NN	O	O
that	NN	O	O
the	NN	O	O
patients	NN	O	B-cell_line
'	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
do	NN	O	O
not	NN	O	O
display	NN	O	O
a	NN	O	O
completely	NN	O	O
``	NN	O	O
bare	NN	O	B-DNA
promoter	NN	O	I-DNA
``	NN	O	O
as	NN	O	O
previously	NN	O	O
reported	NN	O	O
using	NN	O	O
dimethyl	NN	O	O
sulfate	NN	O	O
as	NN	O	O
the	NN	O	O
footprinting	NN	O	O
agent	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
allowed	NN	O	O
us	NN	O	O
,	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
,	NN	O	O
to	NN	O	O
correlate	NN	O	O
the	NN	O	O
AIR-1	NN	O	B-DNA
locus	NN	O	I-DNA
defect	NN	O	I-DNA
with	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
promoter	NN	O	I-DNA
occupancy	NN	O	O
alterations	NN	O	O
and	NN	O	O
distinguish	NN	O	O
these	NN	O	O
alterations	NN	O	O
from	NN	O	O
the	NN	O	O
ones	NN	O	O
observed	NN	O	O
in	NN	O	O
phenotypically	NN	O	O
similar	NN	O	O
but	NN	O	O
genetically	NN	O	O
distinct	NN	O	O
MHC	NN	O	B-cell_line
class	NN	O	I-cell_line
II-negative	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
cell-type-specific	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
alpha-chain	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
:	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
physical	NN	O	O
interactions	NN	O	O
between	NN	O	O
Elf-1	NN	O	B-protein
,	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha-chain	NN	O	I-DNA
(	NN	O	I-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
rapidly	NN	O	O
and	NN	O	O
potently	NN	O	O
induced	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogenic	NN	O	O
stimuli	NN	O	O
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
an	NN	O	O
inducible	NN	O	B-DNA
enhancer	NN	O	I-DNA
between	NN	O	O
nucleotides	NN	O	B-DNA
-299	NN	O	I-DNA
and	NN	O	I-DNA
-228	NN	O	I-DNA
that	NN	O	O
contains	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
CArG	NN	O	I-DNA
motifs	NN	O	I-DNA
was	NN	O	O
identified	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
a	NN	O	O
second	NN	O	O
essential	NN	O	B-DNA
positive	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
located	NN	O	O
between	NN	O	O
nucleotides	NN	O	B-DNA
-137	NN	O	I-DNA
and	NN	O	I-DNA
-64	NN	O	I-DNA
that	NN	O	O
binds	NN	O	O
Elf-1	NN	O	B-protein
and	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

This	NN	O	O
element	NN	O	O
had	NN	O	O
maximal	NN	O	O
activity	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
paralleling	NN	O	O
the	NN	O	O
cell	NN	O	O
type	NN	O	O
specificity	NN	O	O
of	NN	O	O
Elf-1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Transcription	NN	O	O
from	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
was	NN	O	O
inhibited	NN	O	O
when	NN	O	O
either	NN	O	O
the	NN	O	O
Elf-1	NN	O	B-DNA
or	NN	O	I-DNA
the	NN	O	I-DNA
HMG-I	NN	O	I-DNA
(	NN	O	I-DNA
Y	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
mutated	NN	O	O
.	NN	O	O

Coexpression	NN	O	O
of	NN	O	O
both	NN	O	O
proteins	NN	O	O
activated	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
-137	NN	O	B-DNA
to	NN	O	I-DNA
-64	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
COS-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Elf-1	NN	O	B-protein
physically	NN	O	O
associated	NN	O	O
with	NN	O	O
HMG-I	NN	O	B-protein
and	NN	O	O
with	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
might	NN	O	O
functionally	NN	O	O
coordinate	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
and	NN	O	I-DNA
downstream	NN	O	I-DNA
positive	NN	O	I-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
report	NN	O	O
of	NN	O	O
a	NN	O	O
physical	NN	O	O
interaction	NN	O	O
between	NN	O	O
an	NN	O	O
Ets	NN	O	O
family	NN	O	O
member	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
provide	NN	O	O
significant	NN	O	O
new	NN	O	O
insights	NN	O	O
into	NN	O	O
the	NN	O	O
protein-protein	NN	O	O
and	NN	O	O
protein-DNA	NN	O	O
interactions	NN	O	O
that	NN	O	O
regulate	NN	O	O
cell-type-specific	NN	O	O
and	NN	O	O
inducible	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
also	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
other	NN	O	O
genes	NN	O	O
regulated	NN	O	O
by	NN	O	O
Elf-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Isolation	NN	O	O
of	NN	O	O
cDNA	NN	O	B-DNA
clones	NN	O	I-DNA
for	NN	O	O
42	NN	O	O
different	NN	O	O
Kruppel-related	NN	O	B-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
proteins	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monoblast	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U-937	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
complexity	NN	O	O
and	NN	O	O
structural	NN	O	O
characteristics	NN	O	O
of	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
proteins	NN	O	I-protein
expressed	NN	O	O
during	NN	O	O
human	NN	O	O
hematopoiesis	NN	O	O
and	NN	O	O
to	NN	O	O
isolate	NN	O	O
novel	NN	O	O
regulators	NN	O	O
of	NN	O	O
blood	NN	O	O
cell	NN	O	O
development	NN	O	O
,	NN	O	O
a	NN	O	O
degenerate	NN	O	O
oligonucleotide	NN	O	O
probe	NN	O	O
specific	NN	O	O
for	NN	O	O
a	NN	O	O
consensus	NN	O	B-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
peptide	NN	O	I-protein
domain	NN	O	I-protein
was	NN	O	O
used	NN	O	O
to	NN	O	O
isolate	NN	O	O
63	NN	O	O
cDNA	NN	O	B-DNA
clones	NN	O	I-DNA
for	NN	O	O
Kruppel-related	NN	O	B-DNA
zinc	NN	O	I-DNA
finger	NN	O	I-DNA
genes	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monoblast	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U-937	NN	O	I-cell_line
.	NN	O	O

By	NN	O	O
extensive	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
and	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
these	NN	O	O
cDNA	NN	O	B-DNA
clones	NN	O	I-DNA
were	NN	O	O
found	NN	O	O
to	NN	O	O
originate	NN	O	O
from	NN	O	O
approximately	NN	O	O
42	NN	O	O
different	NN	O	O
genes	NN	O	O
(	NN	O	O
HZF	NN	O	B-DNA
1-42	NN	O	I-DNA
)	NN	O	O
of	NN	O	O
which	NN	O	O
only	NN	O	O
8	NN	O	O
have	NN	O	O
previously	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
a	NN	O	O
majority	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
were	NN	O	O
expressed	NN	O	O
at	NN	O	O
comparable	NN	O	O
levels	NN	O	O
in	NN	O	O
U-937	NN	O	B-cell_line
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
individual	NN	O	O
genes	NN	O	O
represented	NN	O	O
among	NN	O	O
the	NN	O	O
63	NN	O	O
clones	NN	O	O
and	NN	O	O
their	NN	O	O
apparent	NN	O	O
non-cell-type-specific	NN	O	O
expression	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
Kruppel-related	NN	O	B-DNA
zinc	NN	O	I-DNA
finger	NN	O	I-DNA
genes	NN	O	I-DNA
are	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
in	NN	O	O
most	NN	O	O
human	NN	O	O
tissues	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
displayed	NN	O	O
a	NN	O	O
restricted	NN	O	O
expression	NN	O	O
pattern	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
they	NN	O	O
represent	NN	O	O
potential	NN	O	O
regulators	NN	O	O
of	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
or	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Detailed	NN	O	O
structural	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
12	NN	O	O
cDNAs	NN	O	B-DNA
(	NN	O	O
HZF	NN	O	B-DNA
1-10	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
a	NN	O	O
partial	NN	O	O
characterization	NN	O	O
of	NN	O	O
HZF	NN	O	B-DNA
11-42	NN	O	I-DNA
revealed	NN	O	O
that	NN	O	O
a	NN	O	O
common	NN	O	O
feature	NN	O	O
of	NN	O	O
human	NN	O	B-protein
Kruppel-related	NN	O	I-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
proteins	NN	O	I-protein
is	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
tandem	NN	O	O
arrays	NN	O	O
of	NN	O	O
zinc	NN	O	B-protein
fingers	NN	O	I-protein
ranging	NN	O	O
in	NN	O	O
number	NN	O	O
from	NN	O	O
3	NN	O	O
to	NN	O	O
over	NN	O	O
20	NN	O	O
that	NN	O	O
are	NN	O	O
preferentially	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
carboxy-terminal	NN	O	B-protein
regions	NN	O	I-protein
of	NN	O	O
the	NN	O	O
proteins	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
several	NN	O	O
novel	NN	O	O
KRAB-containing	NN	O	B-DNA
zinc	NN	O	I-DNA
finger	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
novel	NN	O	O
conserved	NN	O	B-DNA
sequence	NN	O	I-DNA
element	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
.	NN	O	O

-DOCSTART-	O

CAG	NN	O	O
repeat	NN	O	O
length	NN	O	O
variation	NN	O	O
in	NN	O	O
sperm	NN	O	B-cell_type
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
Kennedy	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
modified	NN	O	O
sperm	NN	O	B-cell_type
typing	NN	O	O
protocol	NN	O	O
,	NN	O	O
the	NN	O	O
mutation	NN	O	O
frequency	NN	O	O
of	NN	O	O
the	NN	O	O
CAG	NN	O	B-DNA
repeat	NN	O	I-DNA
region	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
locus	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
measured	NN	O	O
using	NN	O	O
a	NN	O	O
rare	NN	O	O
semen	NN	O	O
sample	NN	O	O
from	NN	O	O
an	NN	O	O
individual	NN	O	O
with	NN	O	O
spinal	NN	O	O
and	NN	O	O
bulbar	NN	O	O
muscular	NN	O	O
atrophy	NN	O	O
(	NN	O	O
SBMA	NN	O	O
)	NN	O	O
.	NN	O	O

Among	NN	O	O
258	NN	O	O
X	NN	O	B-cell_type
chromosome-containing	NN	O	I-cell_type
sperm	NN	O	I-cell_type
,	NN	O	O
19	NN	O	O
%	NN	O	O
had	NN	O	O
a	NN	O	O
repeat	NN	O	O
number	NN	O	O
equal	NN	O	O
to	NN	O	O
the	NN	O	O
donor	NN	O	B-DNA
's	NN	O	I-DNA
somatic	NN	O	I-DNA
DNA	NN	O	I-DNA
(	NN	O	O
47	NN	O	O
repeats	NN	O	O
)	NN	O	O
,	NN	O	O
66	NN	O	O
%	NN	O	O
were	NN	O	O
expansions	NN	O	O
and	NN	O	O
15	NN	O	O
%	NN	O	O
were	NN	O	O
contractions	NN	O	O
.	NN	O	O

The	NN	O	O
average	NN	O	O
expansion	NN	O	O
was	NN	O	O
2.7	NN	O	O
repeats	NN	O	O
.	NN	O	O

More	NN	O	O
than	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
expansions	NN	O	O
involved	NN	O	O
one	NN	O	O
or	NN	O	O
two	NN	O	O
repeats	NN	O	O
;	NN	O	O
the	NN	O	O
largest	NN	O	O
was	NN	O	O
11	NN	O	O
repeats	NN	O	O
.	NN	O	O

68	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
contractions	NN	O	O
were	NN	O	O
also	NN	O	O
one	NN	O	O
or	NN	O	O
two	NN	O	O
repeats	NN	O	O
but	NN	O	O
six	NN	O	O
(	NN	O	O
16	NN	O	O
%	NN	O	O
)	NN	O	O
were	NN	O	O
very	NN	O	O
large	NN	O	O
(	NN	O	O
12-25	NN	O	O
repeats	NN	O	O
)	NN	O	O
.	NN	O	O

One	NN	O	O
contraction	NN	O	O
generated	NN	O	O
an	NN	O	O
allele	NN	O	O
in	NN	O	O
an	NN	O	O
intermediate	NN	O	O
size	NN	O	O
range	NN	O	O
(	NN	O	O
33-39	NN	O	O
repeats	NN	O	O
)	NN	O	O
.	NN	O	O

Such	NN	O	O
alleles	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
observed	NN	O	O
among	NN	O	O
more	NN	O	O
than	NN	O	O
900	NN	O	O
normal	NN	O	O
and	NN	O	O
SBMA	NN	O	B-DNA
X-chromosomes	NN	O	I-DNA
that	NN	O	O
have	NN	O	O
been	NN	O	O
examined	NN	O	O
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
the	NN	O	O
SBMA	NN	O	O
sperm	NN	O	B-cell_type
typing	NN	O	O
results	NN	O	O
with	NN	O	O
mutation	NN	O	O
frequency	NN	O	O
data	NN	O	O
on	NN	O	O
normal	NN	O	O
alleles	NN	O	O
supports	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
trinucleotide	NN	O	B-DNA
repeat	NN	O	I-DNA
expansions	NN	O	O
may	NN	O	O
have	NN	O	O
a	NN	O	O
different	NN	O	O
molecular	NN	O	O
origin	NN	O	O
than	NN	O	O
contractions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Aspirin-like	NN	O	O
drugs	NN	O	O
can	NN	O	O
protect	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
against	NN	O	O
benzoquinone	NN	O	O
cytotoxicity	NN	O	O
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
NAD	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
H	NN	O	I-protein
:	NN	O	I-protein
quinone	NN	O	I-protein
reductase	NN	O	I-protein
-dependent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Benzene	NN	O	O
toxicity	NN	O	O
towards	NN	O	O
lymphocytes	NN	O	B-cell_type
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
metabolites	NN	O	O
of	NN	O	O
benzene	NN	O	O
including	NN	O	O
benzoquinone	NN	O	O
(	NN	O	O
BQ	NN	O	O
)	NN	O	O
.	NN	O	O

NAD	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
H	NN	O	I-protein
:	NN	O	I-protein
quinone	NN	O	I-protein
reductase	NN	O	I-protein
(	NN	O	O
QR	NN	O	B-protein
)	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
protect	NN	O	O
against	NN	O	O
BQ	NN	O	O
toxicity	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
QR	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
had	NN	O	O
previously	NN	O	O
found	NN	O	O
that	NN	O	O
aspirin-like	NN	O	O
drugs	NN	O	O
(	NN	O	O
ALD	NN	O	O
)	NN	O	O
induce	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
was	NN	O	O
therefore	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
ALD	NN	O	O
would	NN	O	O
protect	NN	O	O
lymphocytes	NN	O	B-cell_type
against	NN	O	O
BQ	NN	O	O
toxicity	NN	O	O
by	NN	O	O
inducing	NN	O	O
QR	NN	O	B-protein
.	NN	O	O

Molt-4	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
M4	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
different	NN	O	O
concentrations	NN	O	O
of	NN	O	O
two	NN	O	O
ALD	NN	O	O
,	NN	O	O
flurbiprofen	NN	O	O
and	NN	O	O
sodium	NN	O	O
diclofenac	NN	O	O
,	NN	O	O
and	NN	O	O
then	NN	O	O
exposed	NN	O	O
to	NN	O	O
BQ	NN	O	O
.	NN	O	O

Toxicity	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
viability	NN	O	O
(	NN	O	O
trypan	NN	O	O
blue	NN	O	O
exclusion	NN	O	O
)	NN	O	O
.	NN	O	O

Both	NN	O	O
drugs	NN	O	O
protected	NN	O	O
the	NN	O	O
cells	NN	O	O
against	NN	O	O
BQ	NN	O	O
cytotoxicity	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
e.g.	NN	O	O
,	NN	O	O
sodium	NN	O	O
diclofenac	NN	O	O
at	NN	O	O
15	NN	O	O
microM	NN	O	O
reduced	NN	O	O
the	NN	O	O
fraction	NN	O	O
of	NN	O	O
BQ-treated	NN	O	O
dead	NN	O	O
cells	NN	O	O
by	NN	O	O
70	NN	O	O
%	NN	O	O
.	NN	O	O

ALDs	NN	O	O
induced	NN	O	O
QR	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
the	NN	O	O
M4	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
range	NN	O	O
of	NN	O	O
concentrations	NN	O	O
that	NN	O	O
protected	NN	O	O
the	NN	O	O
cells	NN	O	O
against	NN	O	O
BQ	NN	O	O
toxicity	NN	O	O
.	NN	O	O

The	NN	O	O
protective	NN	O	O
effect	NN	O	O
of	NN	O	O
ALD	NN	O	O
was	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
by	NN	O	O
dicoumarol	NN	O	O
,	NN	O	O
a	NN	O	O
QR	NN	O	B-protein
-specific	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

Since	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
lines	NN	O	I-cell_type
do	NN	O	O
not	NN	O	O
metabolize	NN	O	O
arachidonic	NN	O	O
acid	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
ALD	NN	O	O
can	NN	O	O
protect	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
against	NN	O	O
a	NN	O	O
metabolite	NN	O	O
of	NN	O	O
benzene	NN	O	O
by	NN	O	O
induction	NN	O	O
of	NN	O	O
QR	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Correlation	NN	O	O
of	NN	O	O
differentiation-inducing	NN	O	O
activity	NN	O	O
of	NN	O	O
retinoids	NN	O	O
on	NN	O	O
human	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
HL-60	NN	O	B-cell_line
and	NN	O	O
NB4	NN	O	B-cell_line
.	NN	O	O

Retinoids	NN	O	O
,	NN	O	O
including	NN	O	O
all-trans-retinoic	NN	O	O
acid	NN	O	O
,	NN	O	O
its	NN	O	O
isomers	NN	O	O
,	NN	O	O
and	NN	O	O
fifty	NN	O	O
synthetic	NN	O	O
retinoids	NN	O	O
(	NN	O	O
retinobenzoic	NN	O	O
acids	NN	O	O
)	NN	O	O
,	NN	O	O
were	NN	O	O
tested	NN	O	O
for	NN	O	O
differentiation-inducing	NN	O	O
activity	NN	O	O
on	NN	O	O
human	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
HL-60	NN	O	B-cell_line
and	NN	O	O
NB4	NN	O	B-cell_line
.	NN	O	O

A	NN	O	O
good	NN	O	O
linear	NN	O	O
correlation	NN	O	O
,	NN	O	O
with	NN	O	O
an	NN	O	O
r	NN	O	O
value	NN	O	O
of	NN	O	O
0.91	NN	O	O
,	NN	O	O
between	NN	O	O
the	NN	O	O
ED50	NN	O	O
values	NN	O	O
for	NN	O	O
the	NN	O	O
differentiation-inducing	NN	O	O
activity	NN	O	O
towards	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
that	NN	O	O
towards	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
found	NN	O	O
.	NN	O	O

-DOCSTART-	O

Platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
PAF	NN	O	B-protein
)	NN	O	O
positively	NN	O	O
auto-regulates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-protein
PAF	NN	O	I-protein
receptor	NN	O	I-protein
transcript	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
leukocyte-type	NN	O	O
)	NN	O	O
through	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
platelet-activating	NN	O	I-DNA
factor	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
PAFR	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
transcribed	NN	O	O
by	NN	O	O
two	NN	O	O
distinct	NN	O	O
promoters	NN	O	B-DNA
(	NN	O	O
promoter	NN	O	B-DNA
1	NN	O	I-DNA
and	NN	O	O
promoter	NN	O	B-DNA
2	NN	O	I-DNA
)	NN	O	O
to	NN	O	O
generate	NN	O	O
two	NN	O	O
transcripts	NN	O	O
(	NN	O	O
designated	NN	O	O
as	NN	O	O
PAFR	NN	O	B-RNA
transcript	NN	O	I-RNA
1	NN	O	I-RNA
and	NN	O	O
PAFR	NN	O	B-RNA
transcript	NN	O	I-RNA
2	NN	O	I-RNA
)	NN	O	O
,	NN	O	O
though	NN	O	O
their	NN	O	O
open	NN	O	O
reading	NN	O	O
frames	NN	O	O
are	NN	O	O
identical	NN	O	O
.	NN	O	O

By	NN	O	O
primer	NN	O	O
extension	NN	O	O
analysis	NN	O	O
to	NN	O	O
discriminate	NN	O	O
two	NN	O	O
transcripts	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
PAFR	NN	O	B-RNA
transcript	NN	O	I-RNA
1	NN	O	I-RNA
(	NN	O	O
leukocyte-type	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
PAFR	NN	O	B-RNA
transcript	NN	O	I-RNA
2	NN	O	I-RNA
(	NN	O	O
tissue-type	NN	O	O
)	NN	O	O
,	NN	O	O
are	NN	O	O
upregulated	NN	O	O
by	NN	O	O
PAF	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
stomach	NN	O	I-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
JR-St	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
which	NN	O	O
expresses	NN	O	O
both	NN	O	O
functional	NN	O	O
PAFR	NN	O	B-RNA
transcript	NN	O	I-RNA
1	NN	O	I-RNA
and	NN	O	O
PAFR	NN	O	B-RNA
transcript	NN	O	I-RNA
2	NN	O	I-RNA
endogenously	NN	O	O
.	NN	O	O

Functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
1	NN	O	I-DNA
with	NN	O	O
a	NN	O	O
transient	NN	O	O
expression	NN	O	O
assay	NN	O	O
using	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
reporter	NN	O	O
showed	NN	O	O
that	NN	O	O
both	NN	O	O
PAF	NN	O	B-protein
and	NN	O	O
TPA	NN	O	O
activated	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
1	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
the	NN	O	O
deleted	NN	O	O
promoter	NN	O	O
lacking	NN	O	O
the	NN	O	O
three	NN	O	O
consensus	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
located	NN	O	O
from	NN	O	O
-571	NN	O	O
bp	NN	O	O
to	NN	O	O
-459	NN	O	O
bp	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
a	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
positive	NN	O	O
regulation	NN	O	O
of	NN	O	O
PAFR	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
PAF	NN	O	B-protein
through	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
possibly	NN	O	O
by	NN	O	O
a	NN	O	O
phosphorylation	NN	O	O
reaction	NN	O	O
involving	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
by	NN	O	O
PAF	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

One	NN	O	O
gene	NN	O	O
,	NN	O	O
two	NN	O	O
transcripts	NN	O	B-RNA
:	NN	O	O
isolation	NN	O	O
of	NN	O	O
an	NN	O	O
alternative	NN	O	O
transcript	NN	O	O
encoding	NN	O	O
for	NN	O	O
the	NN	O	O
autoantigen	NN	O	B-protein
La/SS-B	NN	O	I-protein
from	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
library	NN	O	I-DNA
of	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
primary	NN	O	O
Sjogrens	NN	O	O
'	NN	O	O
syndrome	NN	O	O
.	NN	O	O

A	NN	O	O
cDNA	NN	O	B-DNA
library	NN	O	I-DNA
was	NN	O	O
prepared	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
an	NN	O	O
autoimmune	NN	O	O
patient	NN	O	O
with	NN	O	O
primary	NN	O	O
Sjogrens	NN	O	O
'	NN	O	O
syndrome	NN	O	O
.	NN	O	O

The	NN	O	O
cDNA	NN	O	B-DNA
library	NN	O	I-DNA
was	NN	O	O
screened	NN	O	O
with	NN	O	O
the	NN	O	O
patients	NN	O	O
own	NN	O	O
autoimmune	NN	O	O
serum	NN	O	O
being	NN	O	O
monospecific	NN	O	O
for	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
autoantigen	NN	O	I-protein
La/SS-B	NN	O	I-protein
.	NN	O	O

Thereby	NN	O	O
an	NN	O	O
alternative	NN	O	O
type	NN	O	O
of	NN	O	O
La	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
identified	NN	O	O
that	NN	O	O
differed	NN	O	O
from	NN	O	O
the	NN	O	O
known	NN	O	O
La	NN	O	B-RNA
mRNA	NN	O	I-RNA
due	NN	O	O
to	NN	O	O
an	NN	O	O
exchange	NN	O	O
of	NN	O	O
the	NN	O	O
exon	NN	O	B-DNA
1	NN	O	I-DNA
.	NN	O	O

Sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
genomic	NN	O	O
region	NN	O	O
between	NN	O	O
the	NN	O	O
exons	NN	O	B-DNA
1	NN	O	I-DNA
and	NN	O	I-DNA
2	NN	O	I-DNA
showed	NN	O	O
that	NN	O	O
the	NN	O	O
alternative	NN	O	B-DNA
5'-end	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
intron	NN	O	B-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
alternative	NN	O	B-DNA
promoter	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
exists	NN	O	O
within	NN	O	O
the	NN	O	O
intron	NN	O	B-DNA
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
exon	NN	O	B-DNA
1	NN	O	I-DNA
,	NN	O	O
became	NN	O	O
evident	NN	O	O
.	NN	O	O

In	NN	O	O
consequence	NN	O	O
,	NN	O	O
the	NN	O	O
alternative	NN	O	B-RNA
La	NN	O	I-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
promoter	NN	O	O
switching	NN	O	O
combined	NN	O	O
with	NN	O	O
an	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
mechanism	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
intron	NN	O	O
,	NN	O	O
further	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
sites	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
were	NN	O	O
identified	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
suggestion	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
for	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
autoantigen	NN	O	I-protein
La/SS-B	NN	O	I-protein
alters	NN	O	O
in	NN	O	O
dependence	NN	O	O
on	NN	O	O
disease	NN	O	O
conditions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cross-linking	NN	O	O
CD40	NN	O	B-protein
on	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
rapidly	NN	O	O
activates	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
B	NN	O	B-protein
cell-associated	NN	O	I-protein
surface	NN	O	I-protein
molecule	NN	O	I-protein
CD40	NN	O	I-protein
functions	NN	O	O
to	NN	O	O
regulate	NN	O	O
B	NN	O	O
cell	NN	O	O
responses	NN	O	O
.	NN	O	O

Cross-linking	NN	O	O
CD40	NN	O	B-protein
on	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
lead	NN	O	O
to	NN	O	O
homotypic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
,	NN	O	O
IL-6	NN	O	B-protein
production	NN	O	O
,	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
to	NN	O	O
Ig	NN	O	O
isotype	NN	O	O
switching	NN	O	O
.	NN	O	O

Tyrosine	NN	O	O
kinase	NN	O	O
activity	NN	O	O
is	NN	O	O
increased	NN	O	O
shortly	NN	O	O
after	NN	O	O
engagement	NN	O	O
of	NN	O	O
this	NN	O	O
receptor	NN	O	O
.	NN	O	O

Little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
how	NN	O	O
the	NN	O	O
very	NN	O	O
early	NN	O	O
events	NN	O	O
induced	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
cross-linking	NN	O	O
link	NN	O	O
to	NN	O	O
cellular	NN	O	O
responses	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B-like	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
are	NN	O	O
activated	NN	O	O
after	NN	O	O
cross-linking	NN	O	O
CD40	NN	O	B-protein
on	NN	O	O
resting	NN	O	O
human	NN	O	B-cell_type
tonsillar	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
on	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
activation	NN	O	O
is	NN	O	O
rapid	NN	O	O
and	NN	O	O
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
a	NN	O	O
tyrosine	NN	O	O
kinase-dependent	NN	O	O
pathway	NN	O	O
.	NN	O	O

The	NN	O	O
complexes	NN	O	O
detected	NN	O	O
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
contain	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
(	NN	O	O
RelA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
and	NN	O	O
most	NN	O	O
likely	NN	O	O
other	NN	O	O
components	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
cross-linking	NN	O	O
CD40	NN	O	B-protein
supports	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
define	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
system	NN	O	O
as	NN	O	O
an	NN	O	O
intermediate	NN	O	O
event	NN	O	O
in	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
pathway	NN	O	O
can	NN	O	O
influence	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
B	NN	O	B-DNA
cell-associated	NN	O	I-DNA
genes	NN	O	I-DNA
with	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
consensus	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Protease	NN	O	O
inhibitors	NN	O	O
block	NN	O	O
lipopolysaccharide	NN	O	O
induction	NN	O	O
of	NN	O	O
tissue	NN	O	O
factor	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
preventing	NN	O	O
activation	NN	O	O
of	NN	O	O
c-Rel/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
.	NN	O	O

Tissue	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
TF	NN	O	B-protein
)	NN	O	O
is	NN	O	O
expressed	NN	O	O
rapidly	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
exposed	NN	O	O
to	NN	O	O
bacterial	NN	O	B-protein
endotoxin	NN	O	I-protein
(	NN	O	O
lipopolysaccharide	NN	O	O
,	NN	O	O
or	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

Transcriptional	NN	O	O
regulation	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
binding	NN	O	O
of	NN	O	O
c-Rel/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
to	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
B-like	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
TF	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
cytosolic	NN	O	B-protein
c-Rel/p65	NN	O	I-protein
heterodimers	NN	O	I-protein
and	NN	O	O
other	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
family	NN	O	I-protein
requires	NN	O	O
dissociation	NN	O	O
and	NN	O	O
proteolytic	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitor	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
protease	NN	O	O
inhibitors	NN	O	O
N	NN	O	O
alpha-tosylphenylalanyl	NN	O	O
chloromethyl	NN	O	O
ketone	NN	O	O
(	NN	O	O
TPCK	NN	O	O
)	NN	O	O
and	NN	O	O
N	NN	O	O
alpha-tosyl-L-lysine	NN	O	O
chloromethyl	NN	O	O
ketone	NN	O	O
(	NN	O	O
TLCK	NN	O	O
)	NN	O	O
block	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
proteins	NN	O	I-protein
by	NN	O	O
preventing	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

To	NN	O	O
determine	NN	O	O
if	NN	O	O
TPCK	NN	O	O
and	NN	O	O
TLCK	NN	O	O
inhibited	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
monocytic	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
pretreated	NN	O	O
with	NN	O	O
these	NN	O	O
inhibitors	NN	O	O
for	NN	O	O
30	NN	O	O
min	NN	O	O
before	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Both	NN	O	O
TPCK	NN	O	O
and	NN	O	O
TLCK	NN	O	O
inhibited	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
TF	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
TF	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

These	NN	O	O
inhibitors	NN	O	O
specifically	NN	O	O
prevented	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
c-Rel/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
TPCK	NN	O	O
and	NN	O	O
TLCK	NN	O	O
did	NN	O	O
not	NN	O	O
block	NN	O	O
induction	NN	O	O
of	NN	O	O
an	NN	O	O
immediate-early	NN	O	B-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
Egr-1	NN	O	B-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
indicated	NN	O	O
that	NN	O	O
inhibiting	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
c-Rel/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
prevented	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

1	NN	O	B-protein
,	NN	O	I-protein
25-Dihydroxyvitamin	NN	O	I-protein
D3	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
primary	NN	O	O
and	NN	O	O
secondary	NN	O	O
hyperparathyroidism	NN	O	O
.	NN	O	O

A	NN	O	O
decreased	NN	O	O
number	NN	O	O
of	NN	O	O
calcitriol	NN	O	B-protein
(	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
)	NN	O	I-protein
receptors	NN	O	I-protein
has	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
parathyroid	NN	O	O
glands	NN	O	O
of	NN	O	O
uremic	NN	O	O
animals	NN	O	O
.	NN	O	O

In	NN	O	O
humans	NN	O	O
,	NN	O	O
studies	NN	O	O
carried	NN	O	O
out	NN	O	O
in	NN	O	O
surgically	NN	O	O
removed	NN	O	O
parathyroid	NN	O	O
glands	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
calcitriol	NN	O	O
binding	NN	O	O
is	NN	O	O
higher	NN	O	O
in	NN	O	O
primary	NN	O	O
than	NN	O	O
in	NN	O	O
secondary	NN	O	O
hyperparathyroidism	NN	O	O
.	NN	O	O

Since	NN	O	O
specific	NN	O	O
receptors	NN	O	O
for	NN	O	O
calcitriol	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
specific	NN	O	O
uptake	NN	O	O
of	NN	O	O
3H-labelled	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
of	NN	O	O
12	NN	O	O
women	NN	O	O
with	NN	O	O
primary	NN	O	O
hyperparathyroidism	NN	O	O
(	NN	O	O
PHP	NN	O	O
)	NN	O	O
,	NN	O	O
8	NN	O	O
women	NN	O	O
with	NN	O	O
hyperparathyroidism	NN	O	O
secondary	NN	O	O
to	NN	O	O
chronic	NN	O	O
renal	NN	O	O
failure	NN	O	O
(	NN	O	O
SH	NN	O	O
)	NN	O	O
,	NN	O	O
9	NN	O	O
women	NN	O	O
with	NN	O	O
renal	NN	O	O
transplant	NN	O	O
(	NN	O	O
RT	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
23	NN	O	O
healthy	NN	O	O
women	NN	O	O
.	NN	O	O

The	NN	O	O
median	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
was	NN	O	O
similar	NN	O	O
in	NN	O	O
all	NN	O	O
three	NN	O	O
groups	NN	O	O
of	NN	O	O
patients	NN	O	O
and	NN	O	O
in	NN	O	O
healthy	NN	O	O
women	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
S.D.	NN	O	O
(	NN	O	O
range	NN	O	O
)	NN	O	O
:	NN	O	O
PHP	NN	O	O
,	NN	O	O
1.2	NN	O	O
+/-	NN	O	O
1.0	NN	O	O
(	NN	O	O
0.2-4	NN	O	O
)	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
M	NN	O	O
;	NN	O	O
SH	NN	O	O
,	NN	O	O
0.6	NN	O	O
+/-	NN	O	O
0.4	NN	O	O
(	NN	O	O
0.2-1.2	NN	O	O
)	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
M	NN	O	O
;	NN	O	O
RT	NN	O	O
,	NN	O	O
1.1	NN	O	O
+/-	NN	O	O
0.5	NN	O	O
(	NN	O	O
0.4-1.9	NN	O	O
)	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
M	NN	O	O
;	NN	O	O
controls	NN	O	O
,	NN	O	O
1.0	NN	O	O
+/-	NN	O	O
0.6	NN	O	O
(	NN	O	O
0.3-2.6	NN	O	O
)	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
.	NN	O	O
However	NN	O	O
,	NN	O	O
the	NN	O	O
maximal	NN	O	O
binding	NN	O	O
capacity	NN	O	O
(	NN	O	O
Nmax	NN	O	O
)	NN	O	O
was	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
in	NN	O	O
PHP	NN	O	O
(	NN	O	O
3.9	NN	O	O
+/-	NN	O	O
1.9	NN	O	O
(	NN	O	O
1.3-7.6	NN	O	O
)	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
vs.	NN	O	O
2.3	NN	O	O
+/-	NN	O	O
0.9	NN	O	O
(	NN	O	O
1.1-4.4	NN	O	O
)	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
controls	NN	O	O
;	NN	O	O
P	NN	O	O
=	NN	O	O
0.0006	NN	O	O
)	NN	O	O
and	NN	O	O
decreased	NN	O	O
in	NN	O	O
SH	NN	O	O
(	NN	O	O
0.8	NN	O	O
+/-	NN	O	O
0.5	NN	O	O
(	NN	O	O
0.2-1.6	NN	O	O
)	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
vs.	NN	O	O
2.3	NN	O	O
+/-	NN	O	O
0.9	NN	O	O
(	NN	O	O
1.1-4.4	NN	O	O
)	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
controls	NN	O	O
;	NN	O	O
P	NN	O	O
=	NN	O	O
0.0001	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
no	NN	O	O
changes	NN	O	O
were	NN	O	O
seen	NN	O	O
in	NN	O	O
RT	NN	O	O
(	NN	O	O
2.3	NN	O	O
+/-	NN	O	O
0.7	NN	O	O
(	NN	O	O
1.2-3.3	NN	O	O
)	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
vs.	NN	O	O
2.3	NN	O	O
+/-	NN	O	O
0.9	NN	O	O
(	NN	O	O
1.1-4.4	NN	O	O
)	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
controls	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
three	NN	O	O
patients	NN	O	O
with	NN	O	O
PHP	NN	O	O
who	NN	O	O
were	NN	O	O
subjected	NN	O	O
to	NN	O	O
parathyroidectomy	NN	O	O
,	NN	O	O
the	NN	O	O
calcitriol	NN	O	O
number	NN	O	O
came	NN	O	O
down	NN	O	O
to	NN	O	O
normal	NN	O	O
.	NN	O	O

Changes	NN	O	O
of	NN	O	O
calcitriol	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
primary	NN	O	O
and	NN	O	O
secondary	NN	O	O
hyperparathyroidism	NN	O	O
could	NN	O	O
magnify	NN	O	O
the	NN	O	O
consequences	NN	O	O
of	NN	O	O
disturbances	NN	O	O
in	NN	O	O
serum	NN	O	O
concentration	NN	O	O
of	NN	O	O
calcitriol	NN	O	O
itself	NN	O	O
and	NN	O	O
might	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
secondary	NN	O	O
hyperparathyroidism	NN	O	O
in	NN	O	O
uremia	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
of	NN	O	O
females	NN	O	O
with	NN	O	O
tiny	NN	O	B-DNA
ring	NN	O	I-DNA
X	NN	O	I-DNA
chromosomes	NN	O	I-DNA
is	NN	O	O
associated	NN	O	O
with	NN	O	O
inability	NN	O	O
of	NN	O	O
these	NN	O	O
chromosomes	NN	O	B-DNA
to	NN	O	O
undergo	NN	O	O
X	NN	O	O
inactivation	NN	O	O
.	NN	O	O

Mental	NN	O	O
retardation	NN	O	O
and	NN	O	O
a	NN	O	O
constellation	NN	O	O
of	NN	O	O
congenital	NN	O	O
malformations	NN	O	O
not	NN	O	O
usually	NN	O	O
associated	NN	O	O
with	NN	O	O
Turner	NN	O	O
syndrome	NN	O	O
are	NN	O	O
seen	NN	O	O
in	NN	O	O
some	NN	O	O
females	NN	O	O
with	NN	O	O
a	NN	O	O
mosaic	NN	O	O
45	NN	O	O
,	NN	O	O
X/46	NN	O	O
,	NN	O	O
X	NN	O	O
,	NN	O	O
r	NN	O	O
(	NN	O	O
X	NN	O	O
)	NN	O	O
karyotype	NN	O	O
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
these	NN	O	O
females	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
XIST	NN	O	B-DNA
locus	NN	O	I-DNA
on	NN	O	O
their	NN	O	O
tiny	NN	O	B-DNA
ring	NN	O	I-DNA
X	NN	O	I-DNA
chromosomes	NN	O	I-DNA
is	NN	O	O
either	NN	O	O
not	NN	O	O
present	NN	O	O
or	NN	O	O
not	NN	O	O
expressed	NN	O	O
.	NN	O	O

As	NN	O	O
XIST	NN	O	B-DNA
transcription	NN	O	O
is	NN	O	O
well	NN	O	O
correlated	NN	O	O
with	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
chromosome	NN	O	I-DNA
in	NN	O	O
female	NN	O	B-cell_type
somatic	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
spermatogonia	NN	O	B-cell_type
,	NN	O	O
nonexpression	NN	O	O
of	NN	O	O
the	NN	O	O
locus	NN	O	O
even	NN	O	O
when	NN	O	O
it	NN	O	O
is	NN	O	O
present	NN	O	O
suggests	NN	O	O
that	NN	O	O
these	NN	O	O
chromosomes	NN	O	B-DNA
are	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
ring	NN	O	O
X	NN	O	B-DNA
chromosomes	NN	O	I-DNA
lacking	NN	O	O
XIST	NN	O	B-DNA
expression	NN	O	O
(	NN	O	O
XISTE-	NN	O	O
)	NN	O	O
,	NN	O	O
from	NN	O	O
three	NN	O	O
females	NN	O	O
with	NN	O	O
severe	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
tiny	NN	O	O
ring	NN	O	O
X	NN	O	B-DNA
chromosomes	NN	O	I-DNA
studied	NN	O	O
with	NN	O	O
an	NN	O	O
antibody	NN	O	B-protein
specific	NN	O	O
for	NN	O	O
the	NN	O	O
acetylated	NN	O	O
isoforms	NN	O	O
of	NN	O	O
histone	NN	O	B-protein
H4	NN	O	I-protein
marking	NN	O	O
transcribed	NN	O	B-DNA
chromatin	NN	O	I-DNA
domains	NN	O	I-DNA
were	NN	O	O
labeled	NN	O	O
at	NN	O	O
a	NN	O	O
level	NN	O	O
consistent	NN	O	O
with	NN	O	O
their	NN	O	O
being	NN	O	O
active	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
examined	NN	O	O
tow	NN	O	O
of	NN	O	O
the	NN	O	O
XISTE-	NN	O	B-DNA
ring	NN	O	I-DNA
chromosomes	NN	O	I-DNA
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
genes	NN	O	B-DNA
that	NN	O	O
are	NN	O	O
normally	NN	O	O
silent	NN	O	O
on	NN	O	O
an	NN	O	O
inactive	NN	O	B-DNA
X	NN	O	I-DNA
are	NN	O	O
expressed	NN	O	O
from	NN	O	O
these	NN	O	O
chromosomes	NN	O	B-DNA
.	NN	O	O

Analyses	NN	O	O
of	NN	O	O
hybrid	NN	O	B-cell_line
cells	NN	O	I-cell_line
show	NN	O	O
that	NN	O	O
TIMP	NN	O	B-DNA
,	NN	O	O
ZXDA	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
ZXDB	NN	O	B-DNA
loci	NN	O	I-DNA
on	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
short	NN	O	I-DNA
arm	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
AR	NN	O	B-DNA
and	NN	O	O
PHKA1	NN	O	B-DNA
loci	NN	O	I-DNA
on	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
,	NN	O	O
are	NN	O	O
well	NN	O	O
expressed	NN	O	O
from	NN	O	O
the	NN	O	O
tiny	NN	O	B-DNA
ring	NN	O	I-DNA
X	NN	O	I-DNA
chromosome	NN	O	I-DNA
lacking	NN	O	O
XIST	NN	O	B-DNA
DNA	NN	O	I-DNA
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
the	NN	O	O
ring	NN	O	B-DNA
chromosome	NN	O	I-DNA
that	NN	O	O
has	NN	O	O
XIST	NN	O	B-DNA
DNA	NN	O	I-DNA
but	NN	O	O
does	NN	O	O
not	NN	O	O
transcribe	NN	O	O
it	NN	O	O
show	NN	O	O
that	NN	O	O
its	NN	O	O
AR	NN	O	O
allele	NN	O	O
is	NN	O	O
transcribed	NN	O	O
along	NN	O	O
with	NN	O	O
the	NN	O	O
one	NN	O	O
on	NN	O	O
the	NN	O	O
normal	NN	O	O
X	NN	O	O
allele	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
by	NN	O	O
CD28	NN	O	B-protein
signalling	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

Co-stimulation	NN	O	O
of	NN	O	O
T-lymphocytes	NN	O	B-cell_type
by	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
TcR	NN	O	B-protein
)	NN	O	O
occupancy	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
surface	NN	O	I-protein
molecule	NN	O	I-protein
results	NN	O	O
in	NN	O	O
enhanced	NN	O	O
proliferation	NN	O	O
and	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
production	NN	O	O
.	NN	O	O

The	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
triggered	NN	O	O
by	NN	O	O
CD28	NN	O	B-protein
involves	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
B-like	NN	O	I-DNA
sequence	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
5'-regulatory	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
called	NN	O	O
CD28-responsive	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
T-cells	NN	O	B-cell_type
by	NN	O	O
agonistic	NN	O	O
anti-CD28	NN	O	B-protein
antibodies	NN	O	I-protein
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
-	NN	O	O
or	NN	O	O
TcR	NN	O	B-protein
-derived	NN	O	O
signals	NN	O	O
induces	NN	O	O
the	NN	O	O
enhanced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
CD28	NN	O	B-protein
engagement	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
exerts	NN	O	O
opposite	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
.	NN	O	O

Whereas	NN	O	O
anti-CD28	NN	O	B-protein
together	NN	O	O
with	NN	O	O
PMA	NN	O	O
increased	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
trans-activation	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
PMA-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
was	NN	O	O
significantly	NN	O	O
suppressed	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
exerted	NN	O	O
by	NN	O	O
anti-CD28	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
functional	NN	O	O
reporter-gene	NN	O	O
assays	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
independently	NN	O	O
regulated	NN	O	O
and	NN	O	O
may	NN	O	O
perform	NN	O	O
different	NN	O	O
functions	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

HIV-1	NN	O	B-protein
Nef	NN	O	I-protein
leads	NN	O	O
to	NN	O	O
inhibition	NN	O	O
or	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
depending	NN	O	O
on	NN	O	O
its	NN	O	O
intracellular	NN	O	O
localization	NN	O	O
.	NN	O	O

Nef	NN	O	B-protein
of	NN	O	O
primate	NN	O	O
lentiviruses	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
viremia	NN	O	O
and	NN	O	O
progression	NN	O	O
to	NN	O	O
AIDS	NN	O	O
in	NN	O	O
monkeys	NN	O	O
.	NN	O	O

Negative	NN	O	O
,	NN	O	O
positive	NN	O	O
,	NN	O	O
and	NN	O	O
no	NN	O	O
effects	NN	O	O
of	NN	O	O
Nef	NN	O	B-protein
have	NN	O	O
also	NN	O	O
been	NN	O	O
reported	NN	O	O
on	NN	O	O
viral	NN	O	O
replication	NN	O	O
in	NN	O	O
cells	NN	O	O
.	NN	O	O

To	NN	O	O
reconcile	NN	O	O
these	NN	O	O
observations	NN	O	O
,	NN	O	O
we	NN	O	O
expressed	NN	O	O
a	NN	O	O
hybrid	NN	O	B-protein
CD8-Nef	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Two	NN	O	O
opposite	NN	O	O
phenotypes	NN	O	O
were	NN	O	O
found	NN	O	O
,	NN	O	O
which	NN	O	O
depended	NN	O	O
on	NN	O	O
the	NN	O	O
intracellular	NN	O	O
localization	NN	O	O
of	NN	O	O
Nef	NN	O	B-protein
.	NN	O	O

Expressed	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
or	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
,	NN	O	O
the	NN	O	O
chimera	NN	O	B-protein
inhibited	NN	O	O
or	NN	O	O
activated	NN	O	O
early	NN	O	O
signaling	NN	O	O
events	NN	O	O
from	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Activated	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
died	NN	O	O
by	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
and	NN	O	O
only	NN	O	O
cells	NN	O	O
with	NN	O	O
mutated	NN	O	O
nef	NN	O	B-DNA
genes	NN	O	I-DNA
expressing	NN	O	O
truncated	NN	O	O
Nefs	NN	O	B-protein
survived	NN	O	O
,	NN	O	O
which	NN	O	O
rendered	NN	O	O
Nef	NN	O	B-protein
nonfunctional	NN	O	O
.	NN	O	O

These	NN	O	O
mutations	NN	O	O
paralleled	NN	O	O
those	NN	O	O
in	NN	O	O
other	NN	O	O
viral	NN	O	O
strains	NN	O	O
passaged	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Not	NN	O	O
only	NN	O	O
do	NN	O	O
these	NN	O	O
positional	NN	O	O
effects	NN	O	O
of	NN	O	O
Nef	NN	O	B-protein
reconcile	NN	O	O
diverse	NN	O	O
phenotypes	NN	O	O
of	NN	O	O
Nef	NN	O	B-protein
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
its	NN	O	O
N-terminal	NN	O	O
myristylation	NN	O	O
,	NN	O	O
but	NN	O	O
they	NN	O	O
also	NN	O	O
explain	NN	O	O
effects	NN	O	O
of	NN	O	O
Nef	NN	O	B-protein
in	NN	O	O
HIV	NN	O	O
infection	NN	O	O
and	NN	O	O
progression	NN	O	O
to	NN	O	O
AIDS	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
intricate	NN	O	O
arrangement	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
regulates	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
alpha	NN	O	B-DNA
4	NN	O	I-DNA
integrin	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

alpha	NN	O	B-protein
4	NN	O	I-protein
integrins	NN	O	I-protein
mediate	NN	O	O
cell-cell	NN	O	O
and	NN	O	O
cell-extracellular	NN	O	O
matrix	NN	O	O
interactions	NN	O	O
that	NN	O	O
are	NN	O	O
critical	NN	O	O
for	NN	O	O
maturation	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
differentiation	NN	O	O
of	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
examine	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
controlling	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
alpha	NN	O	B-protein
4	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
constructs	NN	O	O
containing	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
deletion	NN	O	I-DNA
mutants	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
alpha	NN	O	B-DNA
4	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
was	NN	O	O
compared	NN	O	O
in	NN	O	O
transfection	NN	O	O
assays	NN	O	O
into	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
express	NN	O	O
alpha	NN	O	B-protein
4	NN	O	I-protein
and	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
do	NN	O	O
not	NN	O	O
.	NN	O	O

The	NN	O	O
sequence	NN	O	O
between	NN	O	O
position	NN	O	O
-42	NN	O	B-DNA
and	NN	O	I-DNA
-76	NN	O	I-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
(	NN	O	O
bp	NN	O	O
)	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
efficient	NN	O	O
transcription	NN	O	O
in	NN	O	O
cells	NN	O	O
that	NN	O	O
express	NN	O	O
alpha	NN	O	B-protein
4	NN	O	I-protein
,	NN	O	O
but	NN	O	O
it	NN	O	O
showed	NN	O	O
no	NN	O	O
activity	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
alpha	NN	O	B-protein
4	NN	O	I-protein
.	NN	O	O

Three	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
found	NN	O	O
in	NN	O	O
this	NN	O	O
region	NN	O	O
:	NN	O	O
two	NN	O	O
adjacent	NN	O	O
sites	NN	O	O
at	NN	O	O
positions	NN	O	O
-50	NN	O	B-DNA
and	NN	O	I-DNA
-54	NN	O	I-DNA
bp	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
more	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
site	NN	O	I-DNA
at	NN	O	O
position	NN	O	O
-67	NN	O	B-DNA
bp	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
constructs	NN	O	O
containing	NN	O	O
deletions	NN	O	O
and	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
3'-most	NN	O	B-DNA
site	NN	O	I-DNA
alone	NN	O	O
was	NN	O	O
sufficient	NN	O	O
for	NN	O	O
binding	NN	O	O
GA-binding	NN	O	B-protein
protein	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
GABP	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
/GABP	NN	O	I-protein
beta	NN	O	I-protein
and	NN	O	O
for	NN	O	O
a	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

When	NN	O	O
all	NN	O	O
three	NN	O	O
sites	NN	O	O
were	NN	O	O
present	NN	O	O
,	NN	O	O
a	NN	O	O
second	NN	O	O
complex	NN	O	O
``	NN	O	O
a	NN	O	O
''	NN	O	O
was	NN	O	O
detected	NN	O	O
,	NN	O	O
which	NN	O	O
contains	NN	O	O
an	NN	O	O
unknown	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

Formation	NN	O	O
of	NN	O	O
complex	NN	O	O
a	NN	O	O
was	NN	O	O
cell-type	NN	O	O
specific	NN	O	O
and	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
the	NN	O	O
5'-most	NN	O	B-DNA
Ets	NN	O	I-DNA
site	NN	O	I-DNA
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
binding	NN	O	O
to	NN	O	O
GABP	NN	O	B-protein
alpha/GABP	NN	O	I-protein
beta	NN	O	I-protein
,	NN	O	O
but	NN	O	O
it	NN	O	O
eliminated	NN	O	O
a	NN	O	O
.	NN	O	O

Concomitant	NN	O	O
with	NN	O	O
this	NN	O	O
loss	NN	O	O
of	NN	O	O
a	NN	O	O
,	NN	O	O
a	NN	O	O
new	NN	O	B-protein
Ets-1-containing	NN	O	I-protein
complex	NN	O	I-protein
``	NN	O	O
c	NN	O	B-protein
``	NN	O	O
appeared	NN	O	O
.	NN	O	O

Complex	NN	O	O
c	NN	O	O
substituted	NN	O	O
efficiently	NN	O	O
for	NN	O	O
complex	NN	O	O
a	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
although	NN	O	O
neither	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
5'-most	NN	O	B-DNA
Ets	NN	O	I-DNA
sites	NN	O	I-DNA
alone	NN	O	O
binds	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
they	NN	O	O
appear	NN	O	O
to	NN	O	O
act	NN	O	O
as	NN	O	O
modulators	NN	O	O
which	NN	O	O
control	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
Ets	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
the	NN	O	O
alpha	NN	O	B-DNA
4	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
arrangement	NN	O	O
of	NN	O	O
Ets	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
coupled	NN	O	O
with	NN	O	O
the	NN	O	O
tissue-	NN	O	O
and	NN	O	O
developmental-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
Ets	NN	O	B-protein
members	NN	O	I-protein
,	NN	O	O
likely	NN	O	O
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
defining	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
alpha	NN	O	B-protein
4	NN	O	I-protein
integrin	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Mechanism	NN	O	O
of	NN	O	O
antiandrogen	NN	O	O
action	NN	O	O
:	NN	O	O
conformational	NN	O	O
changes	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
.	NN	O	O

Androgen	NN	O	B-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
translated	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
and	NN	O	O
androgen-	NN	O	B-protein
and	NN	O	I-protein
antiandrogen-bound	NN	O	I-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
were	NN	O	O
studied	NN	O	O
using	NN	O	O
limited	NN	O	O
proteolytic	NN	O	O
digestion	NN	O	O
by	NN	O	O
trypsin	NN	O	B-protein
.	NN	O	O

Partial	NN	O	O
proteolysis	NN	O	O
of	NN	O	O
androgen-bound	NN	O	B-protein
receptor	NN	O	I-protein
protein	NN	O	I-protein
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
29-kDa	NN	O	B-protein
proteolysis-resisting	NN	O	I-protein
fragment	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
antiandrogen	NN	O	O
binding	NN	O	O
stabilised	NN	O	O
a	NN	O	O
35-kDa	NN	O	B-protein
fragment	NN	O	I-protein
.	NN	O	O

Both	NN	O	O
fragments	NN	O	O
contain	NN	O	O
the	NN	O	O
entire	NN	O	O
ligand	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
35-kDa	NN	O	B-protein
fragment	NN	O	I-protein
extended	NN	O	O
into	NN	O	O
the	NN	O	O
hinge	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
.	NN	O	O

Several	NN	O	O
antiandrogens	NN	O	O
show	NN	O	O
agonistic	NN	O	O
properties	NN	O	O
for	NN	O	O
a	NN	O	O
mutated	NN	O	B-protein
androgen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
LNCaP	NN	O	B-cell_line
cell	NN	O	I-cell_line
variant	NN	O	I-cell_line
)	NN	O	O
;	NN	O	O
trypsin	NN	O	B-protein
digestion	NN	O	O
of	NN	O	O
antiandrogen-bound	NN	O	B-protein
mutated	NN	O	I-protein
receptor	NN	O	I-protein
also	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
29-kDa	NN	O	B-protein
fragment	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
point	NN	O	O
to	NN	O	O
an	NN	O	O
important	NN	O	O
difference	NN	O	O
between	NN	O	O
antiandrogens	NN	O	O
and	NN	O	O
antagonists	NN	O	O
of	NN	O	O
other	NN	O	O
steroid	NN	O	B-protein
hormone	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Antiandrogens	NN	O	O
result	NN	O	O
in	NN	O	O
protection	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
hinge	NN	O	B-protein
region	NN	O	I-protein
and	NN	O	O
C-terminus	NN	O	B-protein
of	NN	O	O
the	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
agonist	NN	O	O
proteolytic	NN	O	O
attack	NN	O	O
,	NN	O	O
whereas	NN	O	O
other	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
antiestrogens	NN	O	O
and	NN	O	O
antiprogestagens	NN	O	O
expose	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
end	NN	O	I-protein
of	NN	O	O
the	NN	O	O
ligand	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
their	NN	O	O
respective	NN	O	O
receptors	NN	O	O
to	NN	O	O
protease	NN	O	B-protein
.	NN	O	O

Differences	NN	O	O
in	NN	O	O
conformation	NN	O	O
of	NN	O	O
the	NN	O	O
hinge	NN	O	B-protein
region	NN	O	I-protein
distinguish	NN	O	O
androgen-bound	NN	O	B-protein
from	NN	O	O
antiandrogen-bound	NN	O	B-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
,	NN	O	O
which	NN	O	O
represents	NN	O	O
an	NN	O	O
important	NN	O	O
feature	NN	O	O
of	NN	O	O
antiandrogen	NN	O	O
action	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
neuroblastoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
eukaryotic	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
it	NN	O	O
is	NN	O	O
constitutively	NN	O	O
present	NN	O	O
in	NN	O	O
cell	NN	O	O
nuclei	NN	O	O
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
many	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocates	NN	O	O
from	NN	O	O
cytosol	NN	O	O
to	NN	O	O
nucleus	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
transduction	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
,	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
,	NN	O	O
and	NN	O	O
other	NN	O	O
polyclonal	NN	O	O
signals	NN	O	O
.	NN	O	O

Using	NN	O	O
neuroblastoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
as	NN	O	O
models	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
in	NN	O	O
neural	NN	O	B-cell_type
cells	NN	O	I-cell_type
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
and	NN	O	O
translocated	NN	O	O
into	NN	O	O
nuclei	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
treatment	NN	O	O
.	NN	O	O

The	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
-activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
transcriptionally	NN	O	O
functional	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
was	NN	O	O
not	NN	O	O
correlated	NN	O	O
with	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
or	NN	O	O
proliferation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
reagents	NN	O	O
such	NN	O	O
as	NN	O	O
nerve	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
NGF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
induce	NN	O	O
phenotypical	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
SH-SY5Y	NN	O	B-cell_line
neuroblastoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
but	NN	O	O
only	NN	O	O
in	NN	O	O
that	NN	O	O
particular	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
a	NN	O	O
NGF-responsive	NN	O	B-cell_line
rat	NN	O	I-cell_line
pheochromocytoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
PC12	NN	O	B-cell_line
,	NN	O	O
PMA	NN	O	O
activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
NGF	NN	O	B-protein
did	NN	O	O
not	NN	O	O
.	NN	O	O

In	NN	O	O
other	NN	O	O
neuroblastoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
such	NN	O	O
as	NN	O	O
SK-N-Be	NN	O	B-cell_line
(	NN	O	I-cell_line
2	NN	O	I-cell_line
)	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
PMA	NN	O	O
induction	NN	O	O
of	NN	O	O
differentiation	NN	O	O
was	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
,	NN	O	O
moreover	NN	O	O
,	NN	O	O
that	NN	O	O
in	NN	O	O
SK-N-Be	NN	O	B-cell_line
(	NN	O	I-cell_line
2	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
was	NN	O	O
much	NN	O	O
lower	NN	O	O
compared	NN	O	O
with	NN	O	O
that	NN	O	O
in	NN	O	O
a	NN	O	O
control	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
that	NN	O	O
the	NN	O	O
low	NN	O	O
PKC	NN	O	B-protein
enzymatic	NN	O	O
activity	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
low	NN	O	O
PKC	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
not	NN	O	O
activated	NN	O	O
by	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
,	NN	O	O
which	NN	O	O
induced	NN	O	O
morphological	NN	O	O
differentiation	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
neuroblastoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
used	NN	O	O
in	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
neuroblastoma	NN	O	B-cell_type
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
obtained	NN	O	O
with	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
proved	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
not	NN	O	O
sufficient	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
neuroblastoma	NN	O	B-cell_type
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

ERP	NN	O	B-protein
,	NN	O	O
a	NN	O	O
new	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
ets	NN	O	B-protein
transcription	NN	O	I-protein
factor/oncoprotein	NN	O	I-protein
family	NN	O	I-protein
:	NN	O	O
cloning	NN	O	O
,	NN	O	O
characterization	NN	O	O
,	NN	O	O
and	NN	O	O
differential	NN	O	O
expression	NN	O	O
during	NN	O	O
B-lymphocyte	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
ets	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
proteins	NN	O	O
which	NN	O	O
function	NN	O	O
as	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
under	NN	O	O
physiological	NN	O	O
conditions	NN	O	O
and	NN	O	O
,	NN	O	O
if	NN	O	O
aberrantly	NN	O	O
expressed	NN	O	O
,	NN	O	O
can	NN	O	O
cause	NN	O	O
cellular	NN	O	O
transformation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
identified	NN	O	O
two	NN	O	O
regulatory	NN	O	O
elements	NN	O	O
in	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
heavy-chain	NN	O	I-DNA
(	NN	O	I-DNA
IgH	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
pi	NN	O	B-DNA
and	NN	O	O
microB	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
exhibit	NN	O	O
striking	NN	O	O
similarity	NN	O	O
to	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
ets-related	NN	O	B-DNA
proteins	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
identify	NN	O	O
ets-related	NN	O	B-protein
transcriptional	NN	O	I-protein
regulators	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
pre-B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
that	NN	O	O
may	NN	O	O
interact	NN	O	O
with	NN	O	O
either	NN	O	O
the	NN	O	O
pi	NN	O	B-DNA
or	NN	O	O
the	NN	O	O
microB	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
have	NN	O	O
used	NN	O	O
a	NN	O	O
PCR	NN	O	O
approach	NN	O	O
with	NN	O	O
degenerate	NN	O	O
oligonucleotides	NN	O	O
encoding	NN	O	O
conserved	NN	O	O
sequences	NN	O	O
in	NN	O	O
all	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
ets	NN	O	B-DNA
family	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
a	NN	O	O
new	NN	O	O
ets-related	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
ERP	NN	O	B-protein
(	NN	O	O
ets-related	NN	O	B-protein
protein	NN	O	I-protein
)	NN	O	O
,	NN	O	O
from	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
pre-B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
BASC	NN	O	B-cell_line
6C2	NN	O	I-cell_line
and	NN	O	O
from	NN	O	O
mouse	NN	O	O
lung	NN	O	O
tissue	NN	O	O
.	NN	O	O

The	NN	O	O
ERP	NN	O	B-protein
protein	NN	O	O
contains	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
high	NN	O	O
homology	NN	O	O
with	NN	O	O
the	NN	O	O
ETS	NN	O	O
DNA-binding	NN	O	O
domain	NN	O	O
common	NN	O	O
to	NN	O	O
all	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
ets	NN	O	B-protein
transcription	NN	O	I-protein
factor/oncoprotein	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Three	NN	O	O
additional	NN	O	O
smaller	NN	O	O
regions	NN	O	O
show	NN	O	O
homology	NN	O	O
to	NN	O	O
the	NN	O	O
ELK-1	NN	O	O
and	NN	O	O
SAP-1	NN	O	B-protein
genes	NN	O	O
,	NN	O	O
a	NN	O	O
subgroup	NN	O	O
of	NN	O	O
the	NN	O	O
ets	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
that	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
serum	NN	O	B-protein
response	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Full-length	NN	O	O
ERP	NN	O	B-protein
expresses	NN	O	O
only	NN	O	O
negligible	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
by	NN	O	O
itself	NN	O	O
.	NN	O	O

Removal	NN	O	O
of	NN	O	O
the	NN	O	O
carboxy	NN	O	B-protein
terminus	NN	O	I-protein
enables	NN	O	O
ERP	NN	O	B-protein
to	NN	O	O
interact	NN	O	O
with	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
ets-binding	NN	O	B-DNA
sites	NN	O	I-DNA
including	NN	O	O
the	NN	O	O
E74	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
pi	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
lck	NN	O	B-DNA
promoter	NN	O	I-DNA
ets	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
carboxy-terminal	NN	O	B-protein
negative	NN	O	I-protein
regulatory	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

At	NN	O	O
least	NN	O	O
three	NN	O	O
ERP-related	NN	O	B-protein
transcripts	NN	O	I-protein
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
tissues	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
within	NN	O	O
the	NN	O	O
B-cell	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
ERP	NN	O	B-protein
is	NN	O	O
highly	NN	O	O
expressed	NN	O	O
primarily	NN	O	O
at	NN	O	O
early	NN	O	O
stages	NN	O	O
of	NN	O	O
B-lymphocyte	NN	O	B-cell_type
development	NN	O	O
,	NN	O	O
and	NN	O	O
expression	NN	O	O
declines	NN	O	O
drastically	NN	O	O
upon	NN	O	O
B-cell	NN	O	O
maturation	NN	O	O
,	NN	O	O
correlating	NN	O	O
with	NN	O	O
the	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
pi	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
ERP	NN	O	B-protein
might	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_type
development	NN	O	O
and	NN	O	O
in	NN	O	O
IgH	NN	O	B-DNA
gene	NN	O	I-DNA
regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Gene	NN	O	O
for	NN	O	O
a	NN	O	O
tissue-specific	NN	O	B-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
(	NN	O	O
EBF	NN	O	B-protein
or	NN	O	O
Olf-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
expressed	NN	O	O
in	NN	O	O
early	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
adipocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
olfactory	NN	O	B-cell_type
neurons	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
located	NN	O	O
on	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
5	NN	O	I-DNA
,	NN	O	O
band	NN	O	B-DNA
q34	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
proximal	NN	O	B-DNA
mouse	NN	O	I-DNA
chromosome	NN	O	I-DNA
11	NN	O	I-DNA
.	NN	O	O

Murine	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
adipocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
olfactory	NN	O	B-cell_type
neurons	NN	O	I-cell_type
contain	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
participates	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
encoding	NN	O	O
tissue-specific	NN	O	O
components	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

Purification	NN	O	O
and	NN	O	O
cloning	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
,	NN	O	O
termed	NN	O	O
early	NN	O	O
B-cell	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
EBF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
from	NN	O	O
murine	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
independent	NN	O	O
cloning	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	O
,	NN	O	O
termed	NN	O	O
Olf-1	NN	O	B-protein
,	NN	O	O
from	NN	O	O
olfactory	NN	O	B-cell_type
neuronal	NN	O	I-cell_type
cells	NN	O	I-cell_type
revealed	NN	O	O
virtual	NN	O	O
complete	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
identity	NN	O	O
between	NN	O	O
these	NN	O	O
proteins	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
first	NN	O	O
step	NN	O	O
towards	NN	O	O
identifying	NN	O	O
a	NN	O	O
human	NN	O	O
genetic	NN	O	O
disorder	NN	O	O
or	NN	O	O
mouse	NN	O	O
mutation	NN	O	O
for	NN	O	O
which	NN	O	O
EBF	NN	O	B-protein
could	NN	O	O
be	NN	O	O
a	NN	O	O
candidate	NN	O	O
gene	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
chromosomally	NN	O	O
mapped	NN	O	O
the	NN	O	O
corresponding	NN	O	O
locus	NN	O	O
in	NN	O	O
both	NN	O	O
species	NN	O	O
.	NN	O	O

By	NN	O	O
Southern	NN	O	O
hybridization	NN	O	O
analyses	NN	O	O
of	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrid	NN	O	O
panels	NN	O	O
with	NN	O	O
murine	NN	O	B-DNA
cDNA	NN	O	I-DNA
probe	NN	O	I-DNA
,	NN	O	O
fluorescence	NN	O	O
chromosomal	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
genomic	NN	O	I-DNA
clones	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
recombinant	NN	O	B-cell_line
inbred	NN	O	I-cell_line
mouse	NN	O	I-cell_line
strains	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
single	NN	O	O
sites	NN	O	O
for	NN	O	O
EBF	NN	O	B-DNA
homologous	NN	O	I-DNA
sequences	NN	O	I-DNA
on	NN	O	O
human	NN	O	O
Chromosome	NN	O	O
(	NN	O	O
Chr	NN	O	O
)	NN	O	O
5	NN	O	O
,	NN	O	O
band	NN	O	B-DNA
q34	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
on	NN	O	O
proximal	NN	O	B-DNA
mouse	NN	O	I-DNA
Chr	NN	O	I-DNA
11	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
an	NN	O	O
evolutionarily	NN	O	B-DNA
conserved	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Calcineurin	NN	O	B-protein
activates	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
synergy	NN	O	O
with	NN	O	O
either	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
or	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
/AP-1	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Two	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
GM-kappa	NN	O	B-DNA
B/GC-box	NN	O	I-DNA
and	NN	O	O
CLE0	NN	O	B-DNA
,	NN	O	O
of	NN	O	O
the	NN	O	O
granulocyte-macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
GM-CSF	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
maximal	NN	O	O
induction	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
costimulation	NN	O	O
with	NN	O	O
phorbol-12-myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
(	NN	O	O
A23187	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
GM-kappa	NN	O	B-DNA
B	NN	O	I-DNA
sequence	NN	O	I-DNA
is	NN	O	O
recognized	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
mainly	NN	O	O
induced	NN	O	O
by	NN	O	O
PMA	NN	O	O
.	NN	O	O

The	NN	O	O
CLE0	NN	O	B-DNA
sequence	NN	O	I-DNA
interacts	NN	O	O
with	NN	O	O
factors	NN	O	O
,	NN	O	O
related	NN	O	O
to	NN	O	O
a	NN	O	O
PMA-induced	NN	O	B-protein
AP-1	NN	O	I-protein
and	NN	O	O
a	NN	O	O
PMA/A23187-induced	NN	O	B-protein
NF-AT	NN	O	I-protein
.	NN	O	O

We	NN	O	O
examined	NN	O	O
whether	NN	O	O
signal	NN	O	O
transducing	NN	O	O
components	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
(	NN	O	I-DNA
p50/p65	NN	O	I-DNA
)	NN	O	I-DNA
-	NN	O	I-DNA
or	NN	O	I-DNA
AP-1	NN	O	I-DNA
(	NN	O	I-DNA
c-Jun/c-Fos	NN	O	I-DNA
)	NN	O	I-DNA
-	NN	O	I-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
into	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
a	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
stimulate	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
cotransfection	NN	O	O
with	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
significantly	NN	O	O
augmented	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
GM-kappa	NN	O	B-DNA
B/GC-box	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
CLE0	NN	O	B-DNA
(	NN	O	O
AP-1/NF-AT	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
a	NN	O	O
constitutively	NN	O	O
active	NN	O	O
calcineurin	NN	O	B-protein
(	NN	O	O
CN	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
Ca2+/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
phosphatase	NN	O	I-protein
,	NN	O	O
potentiated	NN	O	O
by	NN	O	O
two	NN	O	O
fold	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
/AP-1	NN	O	B-protein
.	NN	O	O

Both	NN	O	O
constitutively	NN	O	O
active	NN	O	O
forms	NN	O	O
of	NN	O	O
CN	NN	O	B-protein
and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
synergistically	NN	O	O
activated	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
cooperation	NN	O	O
among	NN	O	O
NF-kappa	NN	O	B-DNA
B-	NN	O	I-DNA
,	NN	O	I-DNA
AP-1-	NN	O	I-DNA
and	NN	O	I-DNA
NF-AT-binding	NN	O	I-DNA
sequences	NN	O	I-DNA
is	NN	O	O
required	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
through	NN	O	O
PKC-	NN	O	O
and	NN	O	O
Ca2+-	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
downstream	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nonpituitary	NN	O	B-DNA
human	NN	O	I-DNA
prolactin	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
Pit-1	NN	O	B-protein
and	NN	O	O
differentially	NN	O	O
controlled	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
in	NN	O	O
endometrial	NN	O	O
stroma	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
PRL	NN	O	I-DNA
(	NN	O	I-DNA
hPRL	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
extrapituitary	NN	O	O
sites	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
uterus	NN	O	O
(	NN	O	O
decidualized	NN	O	O
endometrial	NN	O	O
stroma	NN	O	O
and	NN	O	O
myometrium	NN	O	O
)	NN	O	O
and	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
is	NN	O	O
directed	NN	O	O
by	NN	O	O
an	NN	O	O
alternative	NN	O	B-DNA
promoter	NN	O	I-DNA
which	NN	O	O
is	NN	O	O
located	NN	O	O
approximately	NN	O	O
6	NN	O	B-DNA
kilobases	NN	O	I-DNA
(	NN	O	I-DNA
kb	NN	O	I-DNA
)	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
pituitary-specific	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
delineate	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
mechanisms	NN	O	O
governing	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
nonpituitary	NN	O	O
PRL	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
3	NN	O	B-DNA
kb	NN	O	I-DNA
5'-flanking	NN	O	I-DNA
DNA	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
upstream	NN	O	O
decidual/lymphoid	NN	O	B-DNA
(	NN	O	I-DNA
dPRL	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Based	NN	O	O
on	NN	O	O
sequence	NN	O	O
homology	NN	O	O
we	NN	O	O
identified	NN	O	O
two	NN	O	O
binding	NN	O	B-DNA
motifs	NN	O	I-DNA
for	NN	O	O
Pit-1	NN	O	B-protein
and	NN	O	O
seven	NN	O	O
half-sites	NN	O	B-DNA
for	NN	O	O
glucocorticoid	NN	O	B-protein
receptor/progesterone	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
PR	NN	O	B-protein
)	NN	O	O
binding	NN	O	O
.	NN	O	O

We	NN	O	O
focused	NN	O	O
our	NN	O	O
studies	NN	O	O
on	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Pit-1	NN	O	B-protein
and	NN	O	O
of	NN	O	O
PR	NN	O	B-protein
as	NN	O	O
potential	NN	O	O
transcriptional	NN	O	B-protein
regulators	NN	O	I-protein
,	NN	O	O
since	NN	O	O
the	NN	O	O
POU	NN	O	B-protein
domain	NN	O	I-protein
protein	NN	O	I-protein
Pit-1	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
pituitary	NN	O	O
PRL	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
progesterone	NN	O	O
induces	NN	O	O
decidual	NN	O	O
transformation	NN	O	O
of	NN	O	O
the	NN	O	O
endometrial	NN	O	O
stroma	NN	O	O
,	NN	O	O
a	NN	O	O
differentiation	NN	O	O
process	NN	O	O
during	NN	O	O
which	NN	O	O
the	NN	O	O
decidual	NN	O	O
PRL	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
activated	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
including	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
endometrial	NN	O	O
stroma	NN	O	O
,	NN	O	O
that	NN	O	O
Pit-1	NN	O	B-protein
is	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
dPRL	NN	O	B-DNA
promoter/reporter	NN	O	I-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
carrying	NN	O	O
3	NN	O	B-DNA
kb	NN	O	I-DNA
5'-flanking	NN	O	I-DNA
DNA	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
experiments	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
activated	NN	O	O
PR	NN	O	B-protein
does	NN	O	O
not	NN	O	O
confer	NN	O	O
direct	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
on	NN	O	O
the	NN	O	O
dPRL	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
we	NN	O	O
compared	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transfected	NN	O	O
dPRL	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
PRL-secreting	NN	O	B-cell_type
and	NN	O	I-cell_type
nonsecreting	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
3	NN	O	B-DNA
kb	NN	O	I-DNA
5'-flanking	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
dPRL	NN	O	B-DNA
promoter	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
contain	NN	O	O
elements	NN	O	O
restricting	NN	O	O
expression	NN	O	O
to	NN	O	O
only	NN	O	O
those	NN	O	O
lymphocytes	NN	O	B-cell_type
that	NN	O	O
produce	NN	O	O
PRL	NN	O	O
but	NN	O	O
allowed	NN	O	O
expression	NN	O	O
of	NN	O	O
fusion	NN	O	O
reporter	NN	O	O
genes	NN	O	O
irrespective	NN	O	O
of	NN	O	O
the	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	O
PRL	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
was	NN	O	O
in	NN	O	O
sharp	NN	O	O
contrast	NN	O	O
to	NN	O	O
endometrial	NN	O	O
cells	NN	O	O
where	NN	O	O
3	NN	O	O
kb	NN	O	O
5'-flanking	NN	O	O
DNA	NN	O	O
conferred	NN	O	O
strong	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
on	NN	O	O
the	NN	O	O
dPRL	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
decidualized	NN	O	O
endometrial	NN	O	B-cell_type
stromal	NN	O	I-cell_type
cells	NN	O	I-cell_type
actively	NN	O	O
secreting	NN	O	O
PRL	NN	O	B-protein
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
allow	NN	O	O
transcription	NN	O	O
in	NN	O	O
undifferentiated	NN	O	O
non-PRL-secreting	NN	O	B-cell_type
endometrial	NN	O	I-cell_type
stromal	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
dPRL	NN	O	B-DNA
promoter	NN	O	I-DNA
construct	NN	O	O
in	NN	O	O
these	NN	O	O
undifferentiated	NN	O	O
cells	NN	O	O
could	NN	O	O
however	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
cAMP	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
progesterone	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
a	NN	O	O
signal	NN	O	O
transduced	NN	O	O
through	NN	O	O
the	NN	O	O
cAMP	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
is	NN	O	O
a	NN	O	O
primary	NN	O	O
inducer	NN	O	O
of	NN	O	O
decidual	NN	O	O
PRL	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signals	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
interleukin-4	NN	O	B-DNA
and	NN	O	I-DNA
interleukin-5	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
helper	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Mouse	NN	O	B-cell_line
thymoma	NN	O	I-cell_line
line	NN	O	I-cell_line
EL-4	NN	O	I-cell_line
cells	NN	O	I-cell_line
produce	NN	O	O
cytokines	NN	O	B-protein
such	NN	O	O
as	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
,	NN	O	O
IL-3	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
and	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
.	NN	O	O

EL-4	NN	O	B-cell_line
cells	NN	O	I-cell_line
also	NN	O	O
produce	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
when	NN	O	O
stimulated	NN	O	O
by	NN	O	O
PMA	NN	O	O
alone	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
cAMP	NN	O	O
greatly	NN	O	O
augments	NN	O	O
PMA-dependent	NN	O	O
IL-5	NN	O	B-protein
production	NN	O	O
.	NN	O	O

A	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assay	NN	O	O
revealed	NN	O	O
that	NN	O	O
two	NN	O	O
signals	NN	O	O
,	NN	O	O
PMA	NN	O	O
and	NN	O	O
cAMP	NN	O	O
,	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
cAMP	NN	O	O
almost	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
the	NN	O	O
PMA-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
IL-2	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
transfected	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
positively	NN	O	O
regulated	NN	O	O
by	NN	O	O
cAMP	NN	O	O
in	NN	O	O
a	NN	O	O
manner	NN	O	O
opposite	NN	O	O
to	NN	O	O
that	NN	O	O
for	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
NFs	NN	O	B-protein
)	NN	O	O
that	NN	O	O
regulates	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
promoter	NN	O	I-DNA
to	NN	O	O
cAMP	NN	O	O
and	NN	O	O
PMA	NN	O	O
has	NN	O	O
properties	NN	O	O
similar	NN	O	O
to	NN	O	O
NF	NN	O	B-protein
for	NN	O	O
activated	NN	O	B-cell_type
t	NN	O	I-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
defined	NN	O	O
as	NN	O	O
a	NN	O	O
responsive	NN	O	B-DNA
element	NN	O	I-DNA
for	NN	O	O
PMA	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
(	NN	O	O
A23187	NN	O	O
)	NN	O	O
,	NN	O	O
shares	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
with	NN	O	O
the	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
NF-activated	NN	O	I-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
attempted	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
a	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
for	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
electromobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
both	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
P65	NN	O	B-protein
or	NN	O	O
P65/P50	NN	O	B-protein
heterodimer	NN	O	I-protein
)	NN	O	O
and	NN	O	O
NF-AT	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
sequence	NN	O	O
specificity	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
was	NN	O	O
more	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
and	NN	O	O
only	NN	O	O
a	NN	O	O
small	NN	O	O
amount	NN	O	O
of	NN	O	O
P65	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
a	NN	O	O
component	NN	O	O
or	NN	O	O
components	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
have	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
reconstitute	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
alone	NN	O	O
does	NN	O	O
not	NN	O	O
account	NN	O	O
for	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	O
crude	NN	O	O
extract	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-AT-like	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-4	NN	O	B-DNA
and	NN	O	I-DNA
IL-5	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Solution	NN	O	O
structure	NN	O	O
of	NN	O	O
a	NN	O	O
POU-specific	NN	O	B-protein
homeodomain	NN	O	I-protein
:	NN	O	O
3D-NMR	NN	O	O
studies	NN	O	O
of	NN	O	O
human	NN	O	O
B-cell	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Oct-2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
POU	NN	O	B-protein
DNA-binding	NN	O	I-protein
motif	NN	O	I-protein
defines	NN	O	O
a	NN	O	O
conserved	NN	O	O
family	NN	O	O
of	NN	O	O
eukaryotic	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

This	NN	O	O
bipartite	NN	O	B-protein
motif	NN	O	I-protein
consists	NN	O	O
of	NN	O	O
an	NN	O	O
N-terminal	NN	O	B-protein
POU-specific	NN	O	I-protein
domain	NN	O	I-protein
(	NN	O	O
POUs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
flexible	NN	O	B-protein
linker	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
C-terminal	NN	O	B-protein
POU-specific	NN	O	I-protein
homeodomain	NN	O	I-protein
(	NN	O	O
POUHD	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
solution	NN	O	O
structure	NN	O	O
of	NN	O	O
a	NN	O	O
POU-specific	NN	O	B-protein
homeodomain	NN	O	I-protein
.	NN	O	O

An	NN	O	O
NMR	NN	O	O
model	NN	O	O
is	NN	O	O
obtained	NN	O	O
from	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
a	NN	O	O
human	NN	O	B-protein
B-cell	NN	O	I-protein
specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
which	NN	O	O
participates	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
fragment	NN	O	O
of	NN	O	O
Oct-2	NN	O	B-protein
containing	NN	O	O
POUHD	NN	O	B-protein
and	NN	O	O
an	NN	O	O
adjoining	NN	O	O
linker	NN	O	O
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
and	NN	O	O
characterized	NN	O	O
by	NN	O	O
three-dimensional	NN	O	O
nuclear	NN	O	O
magnetic	NN	O	O
resonance	NN	O	O
(	NN	O	O
3D-NMR	NN	O	O
)	NN	O	O
spectroscopy	NN	O	O
.	NN	O	O

Complete	NN	O	O
1H	NN	O	O
and	NN	O	O
15N	NN	O	O
resonance	NN	O	O
assignment	NN	O	O
of	NN	O	O
the	NN	O	O
POUHD	NN	O	B-protein
moiety	NN	O	I-protein
is	NN	O	O
presented	NN	O	O
.	NN	O	O

The	NN	O	O
POUHD	NN	O	B-protein
solution	NN	O	O
structure	NN	O	O
,	NN	O	O
as	NN	O	O
calculated	NN	O	O
by	NN	O	O
distance	NN	O	O
geometry	NN	O	O
and	NN	O	O
simulated	NN	O	O
annealing	NN	O	O
(	NN	O	O
DG/SA	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
canonical	NN	O	B-protein
homeodomains	NN	O	I-protein
.	NN	O	O

A	NN	O	O
salient	NN	O	O
difference	NN	O	O
between	NN	O	O
solution	NN	O	O
and	NN	O	O
crystal	NN	O	O
structures	NN	O	O
is	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
segment	NN	O	I-protein
of	NN	O	O
alpha-helix	NN	O	B-protein
3	NN	O	I-protein
(	NN	O	O
the	NN	O	O
HTH	NN	O	B-protein
recognition	NN	O	I-protein
helix	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
not	NN	O	O
well	NN	O	O
ordered	NN	O	O
in	NN	O	O
solution	NN	O	O
.	NN	O	O

Because	NN	O	O
this	NN	O	O
segment	NN	O	O
presumably	NN	O	O
folds	NN	O	O
upon	NN	O	O
specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
its	NN	O	O
flexibility	NN	O	O
in	NN	O	O
solution	NN	O	O
may	NN	O	O
reduce	NN	O	O
the	NN	O	O
intrinsic	NN	O	O
DNA	NN	O	O
affinity	NN	O	O
of	NN	O	O
POUHD	NN	O	B-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
POUs	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

NF-kappa	NN	O	O
B-dependent	NN	O	O
and	NN	O	O
-independent	NN	O	O
pathways	NN	O	O
of	NN	O	O
HIV	NN	O	O
activation	NN	O	O
in	NN	O	O
a	NN	O	O
chronically	NN	O	B-cell_line
infected	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

J	NN	O	B-cell_line
delta	NN	O	I-cell_line
K	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
isolated	NN	O	O
as	NN	O	O
a	NN	O	O
chronically	NN	O	B-cell_line
infected	NN	O	I-cell_line
survivor	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
following	NN	O	O
infection	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
dl-NF	NN	O	O
,	NN	O	O
a	NN	O	O
mutated	NN	O	O
strain	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
containing	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
sites	NN	O	O
.	NN	O	O

J	NN	O	B-cell_line
delta	NN	O	I-cell_line
K	NN	O	I-cell_line
cells	NN	O	I-cell_line
exhibited	NN	O	O
very	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
constitutive	NN	O	O
HIV	NN	O	O
production	NN	O	O
.	NN	O	O

HIV-1	NN	O	O
expression	NN	O	O
was	NN	O	O
activated	NN	O	O
from	NN	O	O
J	NN	O	B-cell_line
delta	NN	O	I-cell_line
K	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
,	NN	O	O
sodium	NN	O	O
butyrate	NN	O	O
(	NN	O	O
NaB	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
hexamethylene	NN	O	O
bisacetamide	NN	O	O
(	NN	O	O
HMBA	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
confirming	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
mediating	NN	O	O
TNF-alpha	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
strong	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
by	NN	O	O
NaB	NN	O	O
or	NN	O	O
HMBA	NN	O	O
in	NN	O	O
J	NN	O	B-cell_line
delta	NN	O	I-cell_line
K	NN	O	I-cell_line
cells	NN	O	I-cell_line
clearly	NN	O	O
demonstrates	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-independent	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
HIV	NN	O	O
activation	NN	O	O
in	NN	O	O
chronically	NN	O	B-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

J	NN	O	B-cell_line
delta	NN	O	I-cell_line
K	NN	O	I-cell_line
cells	NN	O	I-cell_line
may	NN	O	O
provide	NN	O	O
a	NN	O	O
useful	NN	O	O
model	NN	O	O
for	NN	O	O
characterizing	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-independent	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

JNK	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
signal	NN	O	O
integration	NN	O	O
during	NN	O	O
costimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
activation	NN	O	O
and	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
production	NN	O	O
require	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
signals	NN	O	O
,	NN	O	O
generated	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
or	NN	O	O
costimulation	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
auxiliary	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
how	NN	O	O
these	NN	O	O
stimuli	NN	O	O
affect	NN	O	O
mitogen	NN	O	B-protein
activated	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MAP	NN	O	I-protein
)	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

Full	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinases	NN	O	I-protein
that	NN	O	O
phosphorylate	NN	O	O
the	NN	O	O
Jun	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
JNK1	NN	O	B-protein
and	NN	O	O
JNK2	NN	O	B-protein
,	NN	O	O
required	NN	O	O
costimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
either	NN	O	O
TPA	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
or	NN	O	O
antibodies	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

Alone	NN	O	O
,	NN	O	O
each	NN	O	O
stimulus	NN	O	O
resulted	NN	O	O
in	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
activation	NN	O	O
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
its	NN	O	O
effect	NN	O	O
on	NN	O	O
IL-2	NN	O	B-protein
induction	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
inhibited	NN	O	O
the	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
JNK	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
competitive	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
Jun	NN	O	B-protein
phosphorylation	NN	O	O
by	NN	O	O
JNK	NN	O	B-protein
inhibited	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinases	NN	O	I-protein
ERK1	NN	O	B-protein
and	NN	O	O
ERK2	NN	O	B-protein
were	NN	O	O
fully	NN	O	O
activated	NN	O	O
by	NN	O	O
TPA	NN	O	O
or	NN	O	O
TCR	NN	O	B-protein
stimulation	NN	O	O
and	NN	O	O
were	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
Ca2+	NN	O	O
,	NN	O	O
CD28	NN	O	O
,	NN	O	O
or	NN	O	O
CsA	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
integration	NN	O	O
of	NN	O	O
signals	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
occurs	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
rat	NN	O	B-cell_type
splenocyte	NN	O	I-cell_type
proliferation	NN	O	O
with	NN	O	O
methylprednisolone	NN	O	O
:	NN	O	O
in	NN	O	O
vivo	NN	O	O
effect	NN	O	O
of	NN	O	O
liposomal	NN	O	O
formulation	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
a	NN	O	O
liposomal	NN	O	O
formulation	NN	O	O
of	NN	O	O
methylprednisolone	NN	O	O
(	NN	O	O
MPL	NN	O	O
)	NN	O	O
on	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
in	NN	O	O
spleen	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
investigated	NN	O	O
following	NN	O	O
IV	NN	O	O
dosing	NN	O	O
in	NN	O	O
rats	NN	O	O
.	NN	O	O

Liposomes	NN	O	O
do	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
suppressive	NN	O	O
action	NN	O	O
of	NN	O	O
MPL	NN	O	O
when	NN	O	O
placed	NN	O	O
in	NN	O	O
lymphocyte	NN	O	B-cell_line
culture	NN	O	I-cell_line
.	NN	O	O

Rat	NN	O	B-cell_type
splenocytes	NN	O	I-cell_type
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
greater	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
MPL	NN	O	O
(	NN	O	O
EC50	NN	O	O
=	NN	O	O
7.9	NN	O	O
nM	NN	O	O
)	NN	O	O
than	NN	O	O
do	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
EC50	NN	O	O
=	NN	O	O
28	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
studies	NN	O	O
in	NN	O	O
rats	NN	O	O
utilized	NN	O	O
2	NN	O	O
mg/kg	NN	O	O
IV	NN	O	O
bolus	NN	O	O
doses	NN	O	O
of	NN	O	O
liposomal	NN	O	O
MPL	NN	O	O
compared	NN	O	O
to	NN	O	O
drug	NN	O	O
in	NN	O	O
solution	NN	O	O
.	NN	O	O

Animals	NN	O	O
were	NN	O	O
sacrificed	NN	O	O
at	NN	O	O
various	NN	O	O
times	NN	O	O
post-dosing	NN	O	O
until	NN	O	O
120	NN	O	O
h	NN	O	O
,	NN	O	O
spleen	NN	O	O
was	NN	O	O
excised	NN	O	O
and	NN	O	O
,	NN	O	O
after	NN	O	O
incubation	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
with	NN	O	O
PHA	NN	O	B-protein
,	NN	O	O
splenocyte	NN	O	O
blastogenic	NN	O	O
responses	NN	O	O
were	NN	O	O
assessed	NN	O	O
by	NN	O	O
measuring	NN	O	O
cellular	NN	O	O
incorporation	NN	O	O
of	NN	O	O
3H-thymidine	NN	O	O
.	NN	O	O

The	NN	O	O
suppressive	NN	O	O
effect	NN	O	O
of	NN	O	O
liposomal	NN	O	O
MPL	NN	O	O
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
free	NN	O	O
drug	NN	O	O
was	NN	O	O
significantly	NN	O	O
prolonged	NN	O	O
(	NN	O	O
>	NN	O	O
120	NN	O	O
h	NN	O	O
vs	NN	O	O
<	NN	O	O
18	NN	O	O
h	NN	O	O
)	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
effects	NN	O	O
versus	NN	O	O
time	NN	O	O
were	NN	O	O
described	NN	O	O
by	NN	O	O
a	NN	O	O
pharmacodynamic	NN	O	O
model	NN	O	O
using	NN	O	O
MPL	NN	O	O
concentrations	NN	O	O
in	NN	O	O
plasma	NN	O	B-cell_type
as	NN	O	O
an	NN	O	O
input	NN	O	O
function	NN	O	O
.	NN	O	O

A	NN	O	O
nonlinear	NN	O	O
relationship	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
suppression	NN	O	O
of	NN	O	O
splenocyte	NN	O	B-cell_type
proliferation	NN	O	O
and	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
bound	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
spleen	NN	O	O
.	NN	O	O

Only	NN	O	O
partial	NN	O	O
receptor	NN	O	O
occupancy	NN	O	O
accompanied	NN	O	O
complete	NN	O	O
lymphocyte	NN	O	O
suppression	NN	O	O
.	NN	O	O

The	NN	O	O
suppression	NN	O	O
of	NN	O	O
endogenous	NN	O	O
corticosterone	NN	O	O
in	NN	O	O
plasma	NN	O	B-cell_type
for	NN	O	O
both	NN	O	O
treatments	NN	O	O
was	NN	O	O
similar	NN	O	O
with	NN	O	O
values	NN	O	O
from	NN	O	O
L-MPL	NN	O	O
rats	NN	O	O
returning	NN	O	O
to	NN	O	O
baseline	NN	O	O
after	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
enhanced	NN	O	O
efficacy	NN	O	O
of	NN	O	O
local	NN	O	O
immunosuppression	NN	O	O
by	NN	O	O
targeting	NN	O	O
spleen	NN	O	O
with	NN	O	O
liposomal	NN	O	O
MPL	NN	O	O
.	NN	O	O

-DOCSTART-	O

Function	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B/Rel	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
invariant	NN	O	I-DNA
chain	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
cell-specific	NN	O	O
binding	NN	O	O
of	NN	O	O
different	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
subunits	NN	O	I-protein
.	NN	O	O

The	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	O
major	NN	O	O
histocompatibility	NN	O	B-DNA
complex	NN	O	I-DNA
class	NN	O	I-DNA
II-associated	NN	O	I-DNA
invariant-chain	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
Ii	NN	O	B-DNA
)	NN	O	O
contains	NN	O	O
two	NN	O	O
NF-kappa	NN	O	B-DNA
B/Rel	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
located	NN	O	O
at	NN	O	O
-109	NN	O	B-DNA
to	NN	O	I-DNA
-118	NN	O	I-DNA
(	NN	O	O
Ii	NN	O	B-DNA
kappa	NN	O	I-DNA
B-1	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
-163	NN	O	B-DNA
to	NN	O	I-DNA
-172	NN	O	I-DNA
(	NN	O	O
Ii	NN	O	B-DNA
kappa	NN	O	I-DNA
B-2	NN	O	I-DNA
)	NN	O	O
from	NN	O	O
the	NN	O	O
transcription	NN	O	O
start	NN	O	O
site	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
differential	NN	O	O
function	NN	O	O
of	NN	O	O
each	NN	O	O
of	NN	O	O
these	NN	O	O
NF-kappa	NN	O	B-DNA
B/Rel	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
several	NN	O	O
distinct	NN	O	O
cell	NN	O	O
types	NN	O	O
depends	NN	O	O
on	NN	O	O
cell-specific	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Ii	NN	O	B-DNA
kappa	NN	O	I-DNA
B-1	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
the	NN	O	O
Ii-expressing	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
H9	NN	O	B-cell_line
,	NN	O	O
but	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
myelomonocytic	NN	O	B-cell_line
and	NN	O	I-cell_line
glia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
protein-DNA	NN	O	O
contacts	NN	O	O
are	NN	O	O
detectable	NN	O	O
at	NN	O	O
Ii	NN	O	B-DNA
kappa	NN	O	I-DNA
B-1	NN	O	I-DNA
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
in	NN	O	O
which	NN	O	O
this	NN	O	O
site	NN	O	O
is	NN	O	O
functional	NN	O	O
as	NN	O	O
either	NN	O	O
a	NN	O	O
positive	NN	O	B-DNA
or	NN	O	I-DNA
negative	NN	O	I-DNA
regulator	NN	O	I-DNA
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
supershift	NN	O	O
assays	NN	O	O
determine	NN	O	O
that	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
can	NN	O	O
bind	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
that	NN	O	O
DNA-binding	NN	O	B-protein
complexes	NN	O	I-protein
that	NN	O	O
contain	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p52	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
cRel	NN	O	B-protein
correlate	NN	O	O
with	NN	O	O
positive	NN	O	O
regulation	NN	O	O
whereas	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
correlates	NN	O	O
with	NN	O	O
negative	NN	O	O
regulation	NN	O	O
.	NN	O	O

Ii	NN	O	B-DNA
kappa	NN	O	I-DNA
B-2	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
site	NN	O	O
of	NN	O	O
positive	NN	O	O
regulation	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
a	NN	O	O
site	NN	O	O
of	NN	O	O
negative	NN	O	O
regulation	NN	O	O
in	NN	O	O
H9	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
myelomonocytic	NN	O	B-cell_line
,	NN	O	I-cell_line
and	NN	O	I-cell_line
glial	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
occupancy	NN	O	O
of	NN	O	O
this	NN	O	O
site	NN	O	O
is	NN	O	O
observed	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
H9	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Again	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
supershift	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p52	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
cRel	NN	O	B-protein
correlates	NN	O	O
with	NN	O	O
positive	NN	O	O
function	NN	O	O
whereas	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
only	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p52	NN	O	B-protein
correlates	NN	O	O
with	NN	O	O
negative	NN	O	O
function	NN	O	O
.	NN	O	O

This	NN	O	O
differential	NN	O	O
binding	NN	O	O
of	NN	O	O
specific	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
subunits	NN	O	I-protein
is	NN	O	O
likely	NN	O	O
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
disparate	NN	O	O
functions	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
NF-kappa	NN	O	B-DNA
B/Rel	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
replicative	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
tumour	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
AIDS-related	NN	O	O
non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
CD4	NN	O	B-protein
cell	NN	O	O
number	NN	O	O
and	NN	O	O
antibody	NN	O	O
titres	NN	O	O
to	NN	O	O
EBV	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
To	NN	O	O
determine	NN	O	O
whether	NN	O	O
activation	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
replication	NN	O	O
in	NN	O	O
tumour	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
AIDS-related	NN	O	O
non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
ARNHL	NN	O	O
)	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
CD4+	NN	O	B-protein
cell	NN	O	O
counts	NN	O	O
and	NN	O	O
influences	NN	O	O
antibody	NN	O	O
response	NN	O	O
to	NN	O	O
EBV	NN	O	O
[	NN	O	O
anti-Z	NN	O	B-protein
Epstein-Barr	NN	O	I-protein
replicative	NN	O	I-protein
activator	NN	O	I-protein
(	NN	O	O
ZEBRA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
anti-early	NN	O	B-protein
antigen	NN	O	I-protein
(	NN	O	O
EA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
anti-viral	NN	O	B-protein
capsid	NN	O	I-protein
antigen	NN	O	I-protein
(	NN	O	O
VCA	NN	O	B-protein
)	NN	O	O
]	NN	O	O
.	NN	O	O

DESIGN	NN	O	O
:	NN	O	O
Retrospective	NN	O	O
study	NN	O	O
based	NN	O	O
on	NN	O	O
immunohistochemistry	NN	O	O
and	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
to	NN	O	O
detect	NN	O	O
EBV	NN	O	O
replicative	NN	O	O
gene	NN	O	O
products	NN	O	O
in	NN	O	O
tissue	NN	O	O
samples	NN	O	O
from	NN	O	O
patients	NN	O	O
affected	NN	O	O
by	NN	O	O
ARNHL	NN	O	O
and	NN	O	O
correlation	NN	O	O
with	NN	O	O
CD4+	NN	O	B-protein
cell	NN	O	O
counts	NN	O	O
and	NN	O	O
results	NN	O	O
of	NN	O	O
EBV	NN	O	O
serology	NN	O	O
(	NN	O	O
including	NN	O	O
anti-	NN	O	O
ZEBRA	NN	O	B-protein
activity	NN	O	O
)	NN	O	O
in	NN	O	O
sera	NN	O	O
from	NN	O	O
the	NN	O	O
same	NN	O	O
patients	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Seventeen	NN	O	O
out	NN	O	O
of	NN	O	O
22	NN	O	O
cases	NN	O	O
of	NN	O	O
ARNHL	NN	O	O
were	NN	O	O
selected	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
EBV	NN	O	O
[	NN	O	O
Epstein-Barr	NN	O	O
early	NN	O	O
region	NN	O	O
(	NN	O	O
EBER	NN	O	O
)	NN	O	O
RNA-positive	NN	O	O
]	NN	O	O
.	NN	O	O

Immunohistochemistry	NN	O	O
was	NN	O	O
performed	NN	O	O
with	NN	O	O
anti-ZEBRA	NN	O	B-protein
,	NN	O	O
anti-EA-restricted	NN	O	B-protein
,	NN	O	I-protein
anti-VCA	NN	O	I-protein
antibodies	NN	O	I-protein
and	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
with	NN	O	O
BHLF1/NotI	NN	O	O
oligoprobes	NN	O	O
on	NN	O	O
tumour	NN	O	O
samples	NN	O	O
.	NN	O	O

Results	NN	O	O
were	NN	O	O
statistically	NN	O	O
correlated	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
CD4+	NN	O	B-protein
cell	NN	O	O
counts	NN	O	O
(	NN	O	O
17	NN	O	O
out	NN	O	O
of	NN	O	O
17	NN	O	O
)	NN	O	O
and	NN	O	O
with	NN	O	O
anti-EBV	NN	O	O
antibody	NN	O	O
titres	NN	O	O
(	NN	O	O
13	NN	O	O
out	NN	O	O
of	NN	O	O
17	NN	O	O
)	NN	O	O
assessed	NN	O	O
using	NN	O	O
standard	NN	O	O
immunofluorescence	NN	O	O
method	NN	O	O
and	NN	O	O
enzyme-linked	NN	O	O
immunosorbent	NN	O	O
assay	NN	O	O
procedure	NN	O	O
using	NN	O	O
recombinant	NN	O	B-protein
ZEBRA	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
as	NN	O	O
antigens	NN	O	B-protein
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
BZLF1	NN	O	B-protein
(	NN	O	O
ZEBRA	NN	O	B-protein
)	NN	O	O
or	NN	O	O
early	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
(	NN	O	O
EA-R	NN	O	B-protein
and	NN	O	O
EA-D/BHLF1/NotI	NN	O	B-protein
)	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
a	NN	O	O
small	NN	O	O
proportion	NN	O	O
(	NN	O	O
<	NN	O	O
0.01-5	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
tumour	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
eight	NN	O	O
of	NN	O	O
these	NN	O	O
17	NN	O	O
cases	NN	O	O
by	NN	O	O
immunohistochemistry	NN	O	O
and	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

Demonstration	NN	O	O
of	NN	O	O
replicative	NN	O	O
gene	NN	O	O
expression	NN	O	O
did	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
either	NN	O	O
low	NN	O	O
CD4+	NN	O	B-protein
cell	NN	O	O
counts	NN	O	O
(	NN	O	O
P	NN	O	O
>	NN	O	O
0.05	NN	O	O
)	NN	O	O
or	NN	O	O
anti-EBV	NN	O	O
antibody	NN	O	O
titres	NN	O	O
(	NN	O	O
P	NN	O	O
>	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

Anti-	NN	O	O
ZEBRA	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
not	NN	O	O
significantly	NN	O	O
increased	NN	O	O
in	NN	O	O
patients	NN	O	O
affected	NN	O	O
with	NN	O	O
ARNHL	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
of	NN	O	O
which	NN	O	O
expressed	NN	O	O
replicative	NN	O	O
gene	NN	O	O
products	NN	O	O
(	NN	O	O
P	NN	O	O
>	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
The	NN	O	O
degree	NN	O	O
of	NN	O	O
immunodeficiency	NN	O	O
does	NN	O	O
not	NN	O	O
clearly	NN	O	O
enhance	NN	O	O
replicative	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
tumour	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
ARNHL	NN	O	O
.	NN	O	O

EBV	NN	O	O
serology	NN	O	O
,	NN	O	O
including	NN	O	O
anti-	NN	O	O
ZEBRA	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
is	NN	O	O
not	NN	O	O
a	NN	O	O
reliable	NN	O	O
tool	NN	O	O
for	NN	O	O
predicting	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
such	NN	O	O
proliferations	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
prostaglandin	NN	O	B-protein
E2	NN	O	I-protein
on	NN	O	O
Th0-type	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
:	NN	O	O
modulation	NN	O	O
of	NN	O	O
functions	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
prostaglandin	NN	O	B-protein
E2	NN	O	I-protein
(	NN	O	O
PGE2	NN	O	B-protein
)	NN	O	O
on	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
and	NN	O	O
proliferation	NN	O	O
of	NN	O	O
the	NN	O	O
CD4+	NN	O	B-cell_line
human	NN	O	I-cell_line
helper	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
SP-B21	NN	O	I-cell_line
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

In	NN	O	O
cells	NN	O	O
stimulated	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
,	NN	O	O
PGE2	NN	O	B-protein
inhibited	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
examined	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
rIL-2	NN	O	B-protein
fully	NN	O	O
restored	NN	O	O
the	NN	O	O
proliferative	NN	O	O
response	NN	O	O
and	NN	O	O
partially	NN	O	O
restored	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-5	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
that	NN	O	O
of	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
in	NN	O	O
cells	NN	O	O
stimulated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
/A23187	NN	O	O
,	NN	O	O
PGE2	NN	O	B-protein
enhanced	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-5	NN	O	B-protein
,	NN	O	O
and	NN	O	O
only	NN	O	O
partially	NN	O	O
inhibited	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
PGE2	NN	O	B-protein
vary	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
mode	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-5	NN	O	B-protein
are	NN	O	O
regulated	NN	O	O
differently	NN	O	O
from	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

In	NN	O	O
a	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
only	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	I-protein
p50/p50	NN	O	I-protein
)	NN	O	I-protein
homodimer	NN	O	I-protein
was	NN	O	O
observed	NN	O	O
in	NN	O	O
a	NN	O	O
complex	NN	O	O
formed	NN	O	O
with	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sequence	NN	O	I-DNA
in	NN	O	O
unstimulated	NN	O	O
SP-B21	NN	O	O
cells	NN	O	O
.	NN	O	O

When	NN	O	O
cells	NN	O	O
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
or	NN	O	O
PMA/A23187	NN	O	O
,	NN	O	O
a	NN	O	O
complex	NN	O	O
formation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
p50	NN	O	B-protein
/p65	NN	O	B-protein
)	NN	O	O
heterodimer	NN	O	O
with	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sequence	NN	O	I-DNA
was	NN	O	O
induced	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
PGE2	NN	O	B-protein
or	NN	O	O
di-butyryl	NN	O	O
(	NN	O	O
Bt2	NN	O	O
)	NN	O	O
cAMP	NN	O	O
abolished	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	I-protein
p50/p65	NN	O	I-protein
)	NN	O	I-protein
heterodimer	NN	O	I-protein
to	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sequence	NN	O	I-DNA
in	NN	O	O
cells	NN	O	O
stimulated	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
but	NN	O	O
not	NN	O	O
with	NN	O	O
PMA/A23187	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
target	NN	O	O
of	NN	O	O
PGE2	NN	O	B-protein
action	NN	O	O
is	NN	O	O
a	NN	O	O
component	NN	O	O
in	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
by	NN	O	O
PGE2	NN	O	B-protein
is	NN	O	O
selective	NN	O	O
.	NN	O	O

PGE2	NN	O	B-protein
enhanced	NN	O	O
the	NN	O	O
complex	NN	O	O
formation	NN	O	O
with	NN	O	O
NF-AT	NN	O	B-DNA
,	NN	O	I-DNA
AP-1	NN	O	I-DNA
and	NN	O	I-DNA
CLE0	NN	O	I-DNA
sequences	NN	O	I-DNA
when	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
activated	NN	O	O
by	NN	O	O
either	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
or	NN	O	O
PMA/A23187	NN	O	O
stimulation	NN	O	O
.	NN	O	O

It	NN	O	O
seems	NN	O	O
therefore	NN	O	O
that	NN	O	O
PGE2	NN	O	B-protein
,	NN	O	O
by	NN	O	O
elevating	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
,	NN	O	O
interferes	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
pathway	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
but	NN	O	O
not	NN	O	O
for	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
or	NN	O	O
CLE0	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
regulatory	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
(	NN	O	O
5'-CGAAAATTTCC-3	NN	O	O
'	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin-4	NN	O	I-DNA
gene	NN	O	I-DNA
:	NN	O	O
relationship	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

The	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin-4	NN	O	I-DNA
(	NN	O	I-DNA
IL-4	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
was	NN	O	O
defined	NN	O	O
as	NN	O	O
a	NN	O	O
responsive	NN	O	B-DNA
element	NN	O	I-DNA
for	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
(	NN	O	O
A23187	NN	O	O
)	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
shares	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
with	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
NF-AT	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
examined	NN	O	O
whether	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
specific	NN	O	O
for	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
P65	NN	O	B-protein
or	NN	O	O
P65/P50	NN	O	B-protein
heterodimer	NN	O	I-protein
)	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
and	NN	O	O
activated	NN	O	O
transcription	NN	O	O
through	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
when	NN	O	O
expression	NN	O	B-DNA
plasmids	NN	O	I-DNA
were	NN	O	O
cotransfected	NN	O	O
with	NN	O	O
P	NN	O	B-DNA
sequence-driven	NN	O	I-DNA
reporter	NN	O	I-DNA
plasmids	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
EMSAs	NN	O	O
,	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
binding	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
unlabeled	NN	O	B-DNA
NF-AT	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
by	NN	O	O
the	NN	O	O
unlabeled	NN	O	B-DNA
AP1	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
purified	NN	O	B-protein
NF-AT	NN	O	I-protein
contained	NN	O	O
an	NN	O	O
activity	NN	O	O
that	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
mobility	NN	O	O
shift	NN	O	O
and	NN	O	O
sequence	NN	O	O
specificity	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
were	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
only	NN	O	O
a	NN	O	O
small	NN	O	O
amount	NN	O	O
of	NN	O	O
P65	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
crude	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
component	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
has	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
reconstitute	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
whereas	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
alone	NN	O	O
can	NN	O	O
not	NN	O	O
account	NN	O	O
for	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
crude	NN	O	O
extracts	NN	O	O
.	NN	O	O

Unlike	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
does	NN	O	O
not	NN	O	O
contain	NN	O	O
AP1	NN	O	B-protein
as	NN	O	O
its	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
component	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
early	NN	O	B-DNA
growth	NN	O	I-DNA
response	NN	O	I-DNA
1	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
and	NN	O	O
pp90rsk	NN	O	B-protein
during	NN	O	O
induction	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
work	NN	O	O
has	NN	O	O
studied	NN	O	O
mechanisms	NN	O	O
responsible	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
early	NN	O	B-DNA
growth	NN	O	I-DNA
response	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	I-DNA
EGR-1	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
U-937	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Differentiation	NN	O	O
of	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
the	NN	O	O
serine/threonine	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
,	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
EGR-1	NN	O	B-DNA
promoter-reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
EGR-1	NN	O	B-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
six	NN	O	O
CC	NN	O	O
(	NN	O	O
A/T	NN	O	O
)	NN	O	O
6GG	NN	O	O
(	NN	O	O
CArG	NN	O	O
)	NN	O	O
motifs	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
5'-most	NN	O	B-DNA
distal	NN	O	I-DNA
CArG	NN	O	I-DNA
sequences	NN	O	I-DNA
conferred	NN	O	O
TPA	NN	O	O
inducibility	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
little	NN	O	O
effect	NN	O	O
of	NN	O	O
TPA	NN	O	O
on	NN	O	O
EGR-1	NN	O	B-DNA
transcription	NN	O	O
in	NN	O	O
a	NN	O	O
TPA-resistant	NN	O	B-cell_line
U-937	NN	O	I-cell_line
cell	NN	O	I-cell_line
variant	NN	O	I-cell_line
,	NN	O	O
designated	NN	O	O
TUR	NN	O	B-cell_line
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
both	NN	O	O
U-937	NN	O	B-cell_line
and	NN	O	O
TUR	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
serine/threonine	NN	O	B-protein
protein	NN	O	I-protein
phosphatases	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	I-protein
2A	NN	O	I-protein
,	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
and	NN	O	O
EGR-1	NN	O	B-DNA
transcription	NN	O	O
through	NN	O	O
the	NN	O	O
5'-most	NN	O	B-DNA
CArG	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Since	NN	O	O
these	NN	O	O
findings	NN	O	O
supported	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
serine/threonine	NN	O	O
protein	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
EGR-1	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
40S	NN	O	B-protein
ribosomal	NN	O	I-protein
protein	NN	O	I-protein
S6	NN	O	I-protein
serine/threonine	NN	O	I-protein
kinases	NN	O	I-protein
,	NN	O	O
pp70S6K	NN	O	B-protein
and	NN	O	O
pp90rsk	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
both	NN	O	O
kinases	NN	O	O
participate	NN	O	O
in	NN	O	O
regulating	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
detectable	NN	O	O
activation	NN	O	O
of	NN	O	O
pp70S6K	NN	O	B-protein
during	NN	O	O
TPA-	NN	O	O
or	NN	O	O
okadaic	NN	O	O
acid-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
rapamycin	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
pp70S6K	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
induction	NN	O	O
of	NN	O	O
EGR-1	NN	O	B-DNA
expression	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
analysis	NN	O	O
of	NN	O	O
pp90rsk	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
a	NN	O	O
peptide	NN	O	O
derived	NN	O	O
from	NN	O	O
S6	NN	O	B-protein
protein	NN	O	I-protein
demonstrated	NN	O	O
stimulation	NN	O	O
of	NN	O	O
this	NN	O	O
kinase	NN	O	B-protein
in	NN	O	O
TPA-treated	NN	O	B-cell_line
U-937	NN	O	I-cell_line
,	NN	O	I-cell_line
and	NN	O	I-cell_line
not	NN	O	I-cell_line
TUR	NN	O	I-cell_line
,	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Okadaic	NN	O	O
acid	NN	O	O
treatment	NN	O	O
of	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
pp90rsk	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Antioxidants	NN	O	O
inhibit	NN	O	O
monocyte	NN	O	B-cell_type
adhesion	NN	O	O
by	NN	O	O
suppressing	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
mobilization	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
to	NN	O	O
generate	NN	O	O
radicals	NN	O	O
.	NN	O	O

Cell	NN	O	O
adhesion	NN	O	O
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	B-protein
by	NN	O	I-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
induction	NN	O	O
of	NN	O	O
surface	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
antioxidant	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
specifically	NN	O	O
inhibits	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Since	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
motifs	NN	O	I-DNA
are	NN	O	O
present	NN	O	O
in	NN	O	O
VCAM-1	NN	O	B-DNA
and	NN	O	I-DNA
intercellular	NN	O	I-DNA
adhesion	NN	O	I-DNA
molecule-1	NN	O	I-DNA
(	NN	O	I-DNA
ICAM-1	NN	O	I-DNA
)	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
used	NN	O	O
PDTC	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
and	NN	O	O
ICAM-1	NN	O	B-protein
induction	NN	O	O
and	NN	O	O
subsequent	NN	O	O
monocyte	NN	O	B-cell_type
adhesion	NN	O	O
in	NN	O	O
TNF-treated	NN	O	B-cell_line
human	NN	O	I-cell_line
umbilical	NN	O	I-cell_line
vein	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
HUVECs	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

PDTC	NN	O	O
or	NN	O	O
N-acetylcysteine	NN	O	O
dose	NN	O	O
dependently	NN	O	O
reduced	NN	O	O
TNF-induced	NN	O	B-protein
VCAM-1	NN	O	I-protein
but	NN	O	O
not	NN	O	O
ICAM-1	NN	O	B-protein
surface	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
also	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
arterial	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
and	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
(	NN	O	O
by	NN	O	O
70	NN	O	O
%	NN	O	O
at	NN	O	O
100	NN	O	O
mumol/L	NN	O	O
PDTC	NN	O	O
)	NN	O	O
in	NN	O	O
HUVECs	NN	O	B-cell_line
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
and	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
.	NN	O	O

Gel-shift	NN	O	O
analysis	NN	O	O
in	NN	O	O
HUVECs	NN	O	B-cell_line
demonstrated	NN	O	O
that	NN	O	O
PDTC	NN	O	O
prevented	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
mobilization	NN	O	O
by	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
only	NN	O	O
VCAM-1	NN	O	B-protein
induction	NN	O	O
was	NN	O	O
controlled	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Since	NN	O	O
HUVECs	NN	O	B-cell_line
released	NN	O	O
superoxide	NN	O	O
anions	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TNF	NN	O	O
,	NN	O	O
and	NN	O	O
H2O2	NN	O	O
induces	NN	O	O
VCAM-1	NN	O	B-protein
,	NN	O	O
PDTC	NN	O	O
may	NN	O	O
act	NN	O	O
as	NN	O	O
a	NN	O	O
radical	NN	O	O
scavenger	NN	O	O
.	NN	O	O

Although	NN	O	O
ICAM-1	NN	O	B-protein
induction	NN	O	O
was	NN	O	O
unaffected	NN	O	O
,	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NADPH	NN	O	B-protein
oxidase	NN	O	I-protein
(	NN	O	O
apocynin	NN	O	O
)	NN	O	O
or	NN	O	O
cytochrome	NN	O	B-protein
P-450	NN	O	I-protein
(	NN	O	O
SKF525a	NN	O	O
)	NN	O	O
suppressed	NN	O	O
VCAM-1	NN	O	B-protein
induction	NN	O	O
by	NN	O	O
TNF	NN	O	O
,	NN	O	O
revealing	NN	O	O
that	NN	O	O
several	NN	O	O
radical-generating	NN	O	O
systems	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
its	NN	O	O
regulation	NN	O	O
.	NN	O	O

PDTC	NN	O	O
,	NN	O	O
apocynin	NN	O	O
,	NN	O	O
or	NN	O	O
SKF525a	NN	O	O
decreased	NN	O	O
adhesion	NN	O	O
of	NN	O	O
monocytic	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
TNF-treated	NN	O	B-cell_line
HUVECs	NN	O	I-cell_line
(	NN	O	O
by	NN	O	O
75	NN	O	O
%	NN	O	O
at	NN	O	O
100	NN	O	O
mumol/L	NN	O	O
PDTC	NN	O	O
)	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
by	NN	O	O
anti-VCAM-1	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
1G11	NN	O	B-protein
indicated	NN	O	O
that	NN	O	O
U937	NN	O	B-cell_line
adhesion	NN	O	O
was	NN	O	O
VCAM-1	NN	O	B-protein
dependent	NN	O	O
and	NN	O	O
suppression	NN	O	O
by	NN	O	O
antioxidants	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
reduced	NN	O	O
VCAM-1	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Displacement	NN	O	O
of	NN	O	O
an	NN	O	O
E-box-binding	NN	O	B-protein
repressor	NN	O	I-protein
by	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
proteins	NN	O	I-protein
:	NN	O	O
implications	NN	O	O
for	NN	O	O
B-cell	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
(	NN	O	I-DNA
IgH	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
although	NN	O	O
it	NN	O	O
binds	NN	O	O
both	NN	O	O
B-cell-restricted	NN	O	B-protein
and	NN	O	I-protein
ubiquitous	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
in	NN	O	O
non-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
upon	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
(	NN	O	I-protein
bHLH	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
E2A	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
not	NN	O	O
only	NN	O	O
by	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
E2A	NN	O	B-protein
to	NN	O	O
its	NN	O	O
cognate	NN	O	O
E	NN	O	B-DNA
box	NN	O	I-DNA
but	NN	O	O
also	NN	O	O
by	NN	O	O
the	NN	O	O
resulting	NN	O	O
displacement	NN	O	O
of	NN	O	O
a	NN	O	O
repressor	NN	O	B-protein
from	NN	O	O
that	NN	O	O
same	NN	O	O
site	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
``	NN	O	B-protein
two-handed	NN	O	I-protein
''	NN	O	I-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
denoted	NN	O	O
ZEB	NN	O	B-protein
,	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
specificity	NN	O	O
of	NN	O	O
which	NN	O	O
mimics	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
repressor	NN	O	I-protein
.	NN	O	O

By	NN	O	O
employing	NN	O	O
a	NN	O	O
derivative	NN	O	O
E	NN	O	B-DNA
box	NN	O	I-DNA
that	NN	O	O
binds	NN	O	O
ZEB	NN	O	B-protein
but	NN	O	O
not	NN	O	O
E2A	NN	O	B-protein
,	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
repressor	NN	O	B-protein
is	NN	O	O
active	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
silenced	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
binding	NN	O	O
competition	NN	O	O
by	NN	O	O
bHLH	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
a	NN	O	O
necessary	NN	O	O
prerequisite	NN	O	O
of	NN	O	O
enhancer	NN	O	B-DNA
activity	NN	O	O
is	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	O
displacement	NN	O	O
of	NN	O	O
a	NN	O	O
ZEB-like	NN	O	B-protein
repressor	NN	O	I-protein
by	NN	O	O
bHLH	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
and	NN	O	O
aspirin	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
multiple	NN	O	O
cellular	NN	O	B-DNA
and	NN	O	I-DNA
viral	NN	O	I-DNA
genes	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
inflammation	NN	O	O
and	NN	O	O
infection	NN	O	O
including	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

The	NN	O	O
anti-inflammatory	NN	O	O
drugs	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
and	NN	O	O
aspirin	NN	O	O
inhibited	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
further	NN	O	O
explains	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
these	NN	O	O
drugs	NN	O	O
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
prevented	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibitor	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
therefore	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
retained	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
.	NN	O	O

Sodium	NN	O	O
salicylate	NN	O	O
and	NN	O	O
aspirin	NN	O	O
also	NN	O	O
inhibited	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
kappa	NN	O	I-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
HIV	NN	O	I-DNA
)	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
transfected	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
the	NN	O	O
antisense	NN	O	O
myb	NN	O	O
expression	NN	O	O
on	NN	O	O
hemin-	NN	O	O
and	NN	O	O
erythropoietin-	NN	O	O
induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
c-myb	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cell	NN	O	I-cell_line
induced	NN	O	O
by	NN	O	O
hemin	NN	O	B-protein
(	NN	O	O
Hm	NN	O	B-protein
)	NN	O	O
and	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
Epo	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
constructed	NN	O	O
recombinant	NN	O	B-DNA
plasmid	NN	O	I-DNA
that	NN	O	O
could	NN	O	O
produce	NN	O	O
antisense	NN	O	B-RNA
myb	NN	O	I-RNA
RNA	NN	O	I-RNA
after	NN	O	O
induction	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

During	NN	O	O
treatment	NN	O	O
with	NN	O	O
Hm	NN	O	B-protein
,	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
constitutively	NN	O	O
expressed	NN	O	O
c-myb	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
them	NN	O	O
began	NN	O	O
to	NN	O	O
synthesize	NN	O	O
hemoglobin	NN	O	B-protein
(	NN	O	O
Hb	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
antisense	NN	O	B-RNA
myb	NN	O	I-RNA
RNA	NN	O	I-RNA
reduced	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
c-myb	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
Hb-synthesizing	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
decreased	NN	O	O
to	NN	O	O
20	NN	O	O
%	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Epo	NN	O	B-protein
,	NN	O	O
c-myb	NN	O	B-RNA
mRNA	NN	O	I-RNA
declined	NN	O	O
and	NN	O	O
20	NN	O	O
%	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
synthesized	NN	O	O
Hb	NN	O	B-protein
regardless	NN	O	O
of	NN	O	O
antisense	NN	O	B-RNA
myb	NN	O	I-RNA
RNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
suggested	NN	O	O
that	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
c-myb	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
Hm	NN	O	B-protein
-induced	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
c-myb	NN	O	B-RNA
mRNA	NN	O	I-RNA
induced	NN	O	O
by	NN	O	O
antisense	NN	O	B-RNA
myb	NN	O	I-RNA
RNA	NN	O	I-RNA
expression	NN	O	O
suppresses	NN	O	O
Hm	NN	O	B-protein
-induced	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
amount	NN	O	O
of	NN	O	O
c-myb	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
reduced	NN	O	O
during	NN	O	O
the	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
Epo	NN	O	B-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
almost	NN	O	O
constant	NN	O	O
during	NN	O	O
Hm	NN	O	B-protein
-induced	NN	O	O
differentiation	NN	O	O
,	NN	O	O
but	NN	O	O
increased	NN	O	O
during	NN	O	O
Epo	NN	O	B-protein
treatment	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
supposed	NN	O	O
that	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
Hm	NN	O	B-protein
-induced	NN	O	O
differentiation	NN	O	O
is	NN	O	O
distinguished	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
Epo	NN	O	B-protein
-induced	NN	O	O
differentiation	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Prenatal	NN	O	O
immune	NN	O	O
challenge	NN	O	O
alters	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenocortical	NN	O	O
axis	NN	O	O
in	NN	O	O
adult	NN	O	O
rats	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
whether	NN	O	O
non-abortive	NN	O	O
maternal	NN	O	O
infections	NN	O	O
would	NN	O	O
compromise	NN	O	O
fetal	NN	O	O
brain	NN	O	O
development	NN	O	O
and	NN	O	O
alter	NN	O	O
hypothalamic-pituitary-adrenocortical	NN	O	O
(	NN	O	O
HPA	NN	O	O
)	NN	O	O
axis	NN	O	O
functioning	NN	O	O
when	NN	O	O
adult	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
putative	NN	O	O
teratogenic	NN	O	O
effects	NN	O	O
of	NN	O	O
a	NN	O	O
T	NN	O	O
cell-mediated	NN	O	O
immune	NN	O	O
response	NN	O	O
versus	NN	O	O
an	NN	O	O
endotoxic	NN	O	O
challenge	NN	O	O
,	NN	O	O
10-d-pregnant	NN	O	O
rats	NN	O	O
received	NN	O	O
a	NN	O	O
single	NN	O	O
intraperitoneal	NN	O	O
injection	NN	O	O
of	NN	O	O
5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
8	NN	O	O
)	NN	O	O
human	NN	O	B-cell_type
red	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HRBC	NN	O	B-cell_type
)	NN	O	O
or	NN	O	O
gram-negative	NN	O	O
bacterial	NN	O	O
endotoxin	NN	O	O
(	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
LPS	NN	O	O
:	NN	O	O
30	NN	O	O
micrograms/kg	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
adult	NN	O	O
male	NN	O	O
progeny	NN	O	O
(	NN	O	O
3	NN	O	O
mo	NN	O	O
old	NN	O	O
)	NN	O	O
of	NN	O	O
both	NN	O	O
experimental	NN	O	O
groups	NN	O	O
showed	NN	O	O
increased	NN	O	O
basal	NN	O	O
plasma	NN	O	O
corticosterone	NN	O	O
levels	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
after	NN	O	O
novelty	NN	O	O
stress	NN	O	O
the	NN	O	O
HRBC	NN	O	B-cell_type
group	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
LPS	NN	O	B-cell_line
group	NN	O	I-cell_line
,	NN	O	O
showed	NN	O	O
increased	NN	O	O
ACTH	NN	O	O
and	NN	O	O
corticosterone	NN	O	O
levels	NN	O	O
.	NN	O	O

Both	NN	O	O
groups	NN	O	O
showed	NN	O	O
substantial	NN	O	O
decreases	NN	O	O
in	NN	O	O
mineralocorticoid	NN	O	O
(	NN	O	O
MR	NN	O	O
)	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
(	NN	O	O
GR	NN	O	O
)	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
hippocampus	NN	O	O
,	NN	O	O
a	NN	O	O
limbic	NN	O	O
brain	NN	O	O
structure	NN	O	O
critical	NN	O	O
for	NN	O	O
HPA	NN	O	O
axis	NN	O	O
regulation	NN	O	O
,	NN	O	O
whereas	NN	O	O
GR	NN	O	B-protein
concentrations	NN	O	O
in	NN	O	O
the	NN	O	O
hypothalamus	NN	O	O
were	NN	O	O
unchanged	NN	O	O
and	NN	O	O
in	NN	O	O
anterior	NN	O	O
pituitary	NN	O	O
were	NN	O	O
slightly	NN	O	O
increased	NN	O	O
.	NN	O	O

HRBC	NN	O	B-cell_type
and	NN	O	O
LPS	NN	O	O
indeed	NN	O	O
stimulated	NN	O	O
the	NN	O	O
maternal	NN	O	O
immune	NN	O	O
system	NN	O	O
as	NN	O	O
revealed	NN	O	O
by	NN	O	O
specific	NN	O	O
anti-HRBC	NN	O	B-protein
antibody	NN	O	I-protein
production	NN	O	O
and	NN	O	O
enhanced	NN	O	O
IL-1	NN	O	O
beta	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
in	NN	O	O
splenocytes	NN	O	B-cell_type
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
a	NN	O	O
T	NN	O	O
cell-mediated	NN	O	O
immune	NN	O	O
response	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
an	NN	O	O
endotoxic	NN	O	O
challenge	NN	O	O
during	NN	O	O
pregnancy	NN	O	O
can	NN	O	O
induce	NN	O	O
anomalies	NN	O	O
in	NN	O	O
HPA	NN	O	O
axis	NN	O	O
function	NN	O	O
in	NN	O	O
adulthood	NN	O	O
.	NN	O	O

Clinically	NN	O	O
,	NN	O	O
it	NN	O	O
may	NN	O	O
be	NN	O	O
postulated	NN	O	O
that	NN	O	O
disturbed	NN	O	O
fetal	NN	O	O
brain	NN	O	O
development	NN	O	O
due	NN	O	O
to	NN	O	O
prenatal	NN	O	O
immune	NN	O	O
challenge	NN	O	O
increases	NN	O	O
the	NN	O	O
vulnerability	NN	O	O
to	NN	O	O
develop	NN	O	O
mental	NN	O	O
illness	NN	O	O
involving	NN	O	O
inadequate	NN	O	O
responses	NN	O	O
to	NN	O	O
stress	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
low	NN	O	O
NM23.H1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
identifying	NN	O	O
high	NN	O	O
malignancy	NN	O	O
human	NN	O	O
melanomas	NN	O	O
.	NN	O	O

The	NN	O	O
NM23	NN	O	B-DNA
gene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
proposed	NN	O	O
as	NN	O	O
a	NN	O	O
metastasis-suppressor	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
its	NN	O	O
use	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
as	NN	O	O
prognostic	NN	O	O
factor	NN	O	O
.	NN	O	O

NM23	NN	O	B-protein
was	NN	O	O
identified	NN	O	O
in	NN	O	O
a	NN	O	O
system	NN	O	O
of	NN	O	O
murine	NN	O	B-cell_line
melanoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
which	NN	O	O
an	NN	O	O
inverse	NN	O	O
relationship	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
NM23	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
metastatic	NN	O	O
ability	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
human	NN	O	O
malignant	NN	O	O
melanoma	NN	O	O
study	NN	O	O
NM23	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
metastases	NN	O	O
that	NN	O	O
developed	NN	O	O
less	NN	O	O
than	NN	O	O
24	NN	O	O
months	NN	O	O
after	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
the	NN	O	O
primary	NN	O	O
tumours	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
paper	NN	O	O
studies	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
NM23.H1	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
which	NN	O	O
derive	NN	O	O
from	NN	O	O
primary	NN	O	O
or	NN	O	O
metastatic	NN	O	O
human	NN	O	O
malignant	NN	O	O
melanomas	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
staging	NN	O	O
,	NN	O	O
infiltration	NN	O	O
degree	NN	O	O
,	NN	O	O
lymphocytic	NN	O	O
infiltration	NN	O	O
,	NN	O	O
cell	NN	O	O
morphology	NN	O	O
,	NN	O	O
cell	NN	O	O
pigmentation	NN	O	O
,	NN	O	O
karyotype	NN	O	O
,	NN	O	O
and	NN	O	O
disease-free	NN	O	O
survival	NN	O	O
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
NM23	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
derive	NN	O	O
from	NN	O	O
more	NN	O	O
infiltrating	NN	O	O
primary	NN	O	O
melanomas	NN	O	O
than	NN	O	O
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
obtained	NN	O	O
from	NN	O	O
less	NN	O	O
infiltrating	NN	O	O
tumours	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
tumours	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
disease-free	NN	O	O
survival	NN	O	O
of	NN	O	O
more	NN	O	O
than	NN	O	O
24	NN	O	O
months	NN	O	O
(	NN	O	O
24-58	NN	O	O
months	NN	O	O
)	NN	O	O
express	NN	O	O
the	NN	O	O
NM23	NN	O	B-DNA
gene	NN	O	I-DNA
at	NN	O	O
higher	NN	O	O
levels	NN	O	O
than	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
obtained	NN	O	O
from	NN	O	O
melanomas	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
disease-free	NN	O	O
survival	NN	O	O
of	NN	O	O
less	NN	O	O
than	NN	O	O
24	NN	O	O
months	NN	O	O
(	NN	O	O
6-15	NN	O	O
months	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
serine/threonine	NN	O	B-protein
kinase	NN	O	I-protein
that	NN	O	O
phosphorylates	NN	O	O
c-Fos	NN	O	B-protein
upon	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
via	NN	O	O
antigen	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Ligation	NN	O	O
of	NN	O	O
Ag	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
initiates	NN	O	O
signal	NN	O	O
transduction	NN	O	O
cascades	NN	O	O
which	NN	O	O
alter	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
that	NN	O	O
regulate	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
transmission	NN	O	O
of	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
membrane	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
is	NN	O	O
mediated	NN	O	O
principally	NN	O	O
through	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
and	NN	O	I-protein
serine/threonine	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
and	NN	O	O
characterized	NN	O	O
a	NN	O	O
novel	NN	O	O
serine/threonine	NN	O	B-protein
kinase	NN	O	I-protein
that	NN	O	O
phosphorylated	NN	O	O
the	NN	O	O
proto-oncogene	NN	O	B-protein
product	NN	O	I-protein
,	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
is	NN	O	O
termed	NN	O	O
Fos	NN	O	B-protein
kinase	NN	O	I-protein
.	NN	O	O

Fos	NN	O	B-protein
kinase	NN	O	I-protein
was	NN	O	O
rapidly	NN	O	O
activated	NN	O	O
after	NN	O	O
ligation	NN	O	O
of	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
and	NN	O	I-protein
CD2	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
and	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
anti-IgM	NN	O	B-protein
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
AF10	NN	O	B-cell_line
and	NN	O	O
Ramos	NN	O	B-cell_line
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
,	NN	O	O
PMA	NN	O	O
,	NN	O	O
was	NN	O	O
also	NN	O	O
a	NN	O	O
potent	NN	O	O
inducer	NN	O	O
of	NN	O	O
Fos	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
above	NN	O	O
populations	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
PKC	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
this	NN	O	O
enzyme	NN	O	B-protein
.	NN	O	O

Fos	NN	O	B-protein
kinase	NN	O	I-protein
phosphorylates	NN	O	O
c-Fos	NN	O	B-protein
at	NN	O	O
a	NN	O	O
site	NN	O	O
near	NN	O	O
the	NN	O	O
C-terminus	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
peptide	NN	O	O
derived	NN	O	O
from	NN	O	O
this	NN	O	O
region	NN	O	O
(	NN	O	O
residues	NN	O	B-protein
359-370	NN	O	I-protein
,	NN	O	O
RKGSSSNEPSSD	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
Fos	NN	O	O
peptide	NN	O	O
competitively	NN	O	O
inhibited	NN	O	O
c-Fos	NN	O	B-protein
phosphorylation	NN	O	O
.	NN	O	O

Fos	NN	O	B-protein
kinase	NN	O	I-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
distinct	NN	O	O
from	NN	O	O
other	NN	O	O
identified	NN	O	O
serine/threonine	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
including	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
,	NN	O	O
casein	NN	O	B-protein
kinase	NN	O	I-protein
II	NN	O	I-protein
,	NN	O	O
MAP	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
p70S6K	NN	O	B-protein
and	NN	O	O
p90RSK	NN	O	B-protein
.	NN	O	O

Fos	NN	O	B-protein
kinase	NN	O	I-protein
was	NN	O	O
purified	NN	O	O
by	NN	O	O
anion	NN	O	O
exchange	NN	O	O
chromatography	NN	O	O
and	NN	O	O
exhibited	NN	O	O
an	NN	O	O
apparent	NN	O	O
M	NN	O	O
(	NN	O	O
r	NN	O	O
)	NN	O	O
=	NN	O	O
65	NN	O	O
,	NN	O	O
000	NN	O	O
and	NN	O	O
isoelectric	NN	O	O
point	NN	O	O
=	NN	O	O
6.1	NN	O	O
.	NN	O	O

Fos	NN	O	B-protein
kinase	NN	O	I-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
through	NN	O	O
its	NN	O	O
capacity	NN	O	O
to	NN	O	O
phosphorylate	NN	O	O
c-Fos	NN	O	B-protein
at	NN	O	O
a	NN	O	O
site	NN	O	O
required	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
transrepressive	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
molecule	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
its	NN	O	O
rapid	NN	O	O
activation	NN	O	O
suggests	NN	O	O
it	NN	O	O
may	NN	O	O
have	NN	O	O
a	NN	O	O
wider	NN	O	O
role	NN	O	O
within	NN	O	O
signal	NN	O	O
transduction	NN	O	O
cascades	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Antigenic	NN	O	O
specificities	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
recovered	NN	O	O
from	NN	O	O
recurrent	NN	O	O
genital	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
virus	NN	O	O
type	NN	O	O
2	NN	O	O
lesions	NN	O	O
.	NN	O	O

Lesions	NN	O	O
resulting	NN	O	O
from	NN	O	O
recurrent	NN	O	O
genital	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
virus	NN	O	O
(	NN	O	O
HSV	NN	O	O
)	NN	O	O
infection	NN	O	O
are	NN	O	O
characterized	NN	O	O
by	NN	O	O
infiltration	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
antigenic	NN	O	O
specificity	NN	O	O
of	NN	O	O
47	NN	O	O
HSV-specific	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
recovered	NN	O	O
from	NN	O	O
the	NN	O	O
HSV-2	NN	O	O
buttock	NN	O	O
and	NN	O	O
thigh	NN	O	O
lesions	NN	O	O
of	NN	O	O
five	NN	O	O
patients	NN	O	O
.	NN	O	O

Clones	NN	O	O
with	NN	O	O
proliferative	NN	O	O
responses	NN	O	O
to	NN	O	O
recombinant	NN	O	B-protein
truncated	NN	O	I-protein
glycoprotein	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
gB	NN	O	B-protein
)	NN	O	O
or	NN	O	O
gD	NN	O	B-protein
of	NN	O	O
HSV-2	NN	O	O
or	NN	O	O
purified	NN	O	O
natural	NN	O	O
gC	NN	O	O
of	NN	O	O
HSV-2	NN	O	O
comprised	NN	O	O
a	NN	O	O
minority	NN	O	O
of	NN	O	O
the	NN	O	O
total	NN	O	O
number	NN	O	O
of	NN	O	O
HSV-specific	NN	O	B-cell_line
clones	NN	O	I-cell_line
isolated	NN	O	O
from	NN	O	O
lesions	NN	O	O
.	NN	O	O

The	NN	O	O
gC2-	NN	O	B-cell_line
and	NN	O	I-cell_line
gD2-specific	NN	O	I-cell_line
CD4+	NN	O	I-cell_line
clones	NN	O	I-cell_line
had	NN	O	O
cytotoxic	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
approximate	NN	O	O
locations	NN	O	O
of	NN	O	O
the	NN	O	O
HSV-2	NN	O	O
genes	NN	O	O
encoding	NN	O	O
HSV-2	NN	O	O
type-specific	NN	O	O
CD4+	NN	O	B-protein
antigens	NN	O	I-protein
have	NN	O	O
been	NN	O	O
determined	NN	O	O
by	NN	O	O
using	NN	O	O
HSV-1	NN	O	O
x	NN	O	O
HSV-2	NN	O	O
intertypic	NN	O	O
recombinant	NN	O	O
virus	NN	O	O
and	NN	O	O
include	NN	O	O
the	NN	O	O
approximate	NN	O	O
map	NN	O	O
regions	NN	O	O
0.30	NN	O	O
to	NN	O	O
0.46	NN	O	O
,	NN	O	O
0.59	NN	O	O
to	NN	O	O
0.67	NN	O	O
,	NN	O	O
0.67	NN	O	O
to	NN	O	O
0.73	NN	O	O
,	NN	O	O
and	NN	O	O
0.82	NN	O	O
to	NN	O	O
1.0	NN	O	O
units	NN	O	O
.	NN	O	O

The	NN	O	O
antigenic	NN	O	O
specificity	NN	O	O
of	NN	O	O
an	NN	O	O
HLA	NN	O	B-cell_line
DQ2-restricted	NN	O	I-cell_line
,	NN	O	I-cell_line
HSV-2	NN	O	I-cell_line
type-specific	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
425	NN	O	I-protein
to	NN	O	I-protein
444	NN	O	I-protein
of	NN	O	O
VP16	NN	O	B-protein
of	NN	O	O
HSV-2	NN	O	O
by	NN	O	O
sequential	NN	O	O
use	NN	O	O
of	NN	O	O
an	NN	O	O
intertypic	NN	O	O
recombinant	NN	O	O
virus	NN	O	O
containing	NN	O	O
VP16	NN	O	B-protein
of	NN	O	O
HSV-2	NN	O	O
in	NN	O	O
an	NN	O	O
HSV-1	NN	O	O
background	NN	O	O
,	NN	O	O
recombinant	NN	O	B-protein
VP16	NN	O	I-protein
fusion	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
and	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
.	NN	O	O

Each	NN	O	O
of	NN	O	O
the	NN	O	O
remaining	NN	O	O
four	NN	O	O
patients	NN	O	O
also	NN	O	O
yielded	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
type-specific	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
reactive	NN	O	O
with	NN	O	O
an	NN	O	O
HSV-2	NN	O	O
epitope	NN	O	O
mapping	NN	O	O
to	NN	O	O
approximately	NN	O	O
0.67	NN	O	O
to	NN	O	O
0.73	NN	O	O
map	NN	O	O
units	NN	O	O
.	NN	O	O

The	NN	O	O
antigenic	NN	O	O
specificities	NN	O	O
of	NN	O	O
lesion-derived	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
are	NN	O	O
quite	NN	O	O
diverse	NN	O	O
and	NN	O	O
include	NN	O	O
at	NN	O	O
least	NN	O	O
10	NN	O	O
epitopes	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
reactive	NN	O	O
with	NN	O	O
gC	NN	O	B-protein
and	NN	O	O
VP16	NN	O	B-protein
are	NN	O	O
reported	NN	O	O
here	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
.	NN	O	O

-DOCSTART-	O

Marked	NN	O	O
basophilia	NN	O	O
in	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukaemia	NN	O	O
treated	NN	O	O
with	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
:	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
basophils	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
report	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukaemia	NN	O	O
who	NN	O	O
developed	NN	O	O
marked	NN	O	O
basophilia	NN	O	O
during	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
treatment	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
genomic	NN	O	B-DNA
DNA	NN	O	I-DNA
and	NN	O	O
RNA	NN	O	O
extracted	NN	O	O
from	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
peripheral	NN	O	B-cell_type
leucocytes	NN	O	I-cell_type
in	NN	O	O
order	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
basophils	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
rearranged	NN	O	O
band	NN	O	O
in	NN	O	O
the	NN	O	O
Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
and	NN	O	O
a	NN	O	O
chimaeric	NN	O	B-protein
product	NN	O	I-protein
of	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
were	NN	O	O
strongly	NN	O	O
visible	NN	O	O
before	NN	O	O
ATRA	NN	O	O
treatment	NN	O	O
,	NN	O	O
but	NN	O	O
at	NN	O	O
the	NN	O	O
time	NN	O	O
of	NN	O	O
maximal	NN	O	O
basophilia	NN	O	O
both	NN	O	O
of	NN	O	O
them	NN	O	O
were	NN	O	O
markedly	NN	O	O
diminished	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
basophils	NN	O	B-cell_type
which	NN	O	O
appeared	NN	O	O
during	NN	O	O
the	NN	O	O
ATRA	NN	O	O
treatment	NN	O	O
are	NN	O	O
reactive	NN	O	O
in	NN	O	O
nature	NN	O	O
rather	NN	O	O
than	NN	O	O
a	NN	O	O
leukaemic	NN	O	B-cell_line
clone	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
6	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
Mycobacterium	NN	O	O
tuberculosis	NN	O	O
or	NN	O	O
lipopolysaccharide	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
NF-IL6	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
Proc	NN	O	O
Natl	NN	O	O
Acad	NN	O	O
Sci	NN	O	O
U	NN	O	O
S	NN	O	O
A	NN	O	O
1995	NN	O	O
Apr	NN	O	O
11	NN	O	O
;	NN	O	O
92	NN	O	O
(	NN	O	O
8	NN	O	O
)	NN	O	O
:	NN	O	O
3632	NN	O	O
]	NN	O	O

The	NN	O	O
host	NN	O	O
response	NN	O	O
to	NN	O	O
Mycobacterium	NN	O	O
tuberculosis	NN	O	O
includes	NN	O	O
granuloma	NN	O	O
formation	NN	O	O
at	NN	O	O
sites	NN	O	O
of	NN	O	O
infection	NN	O	O
and	NN	O	O
systemic	NN	O	O
symptoms	NN	O	O
.	NN	O	O

Cytokines	NN	O	B-protein
have	NN	O	O
been	NN	O	O
identified	NN	O	O
by	NN	O	O
immunohistochemistry	NN	O	O
in	NN	O	O
granulomas	NN	O	O
in	NN	O	O
animal	NN	O	O
models	NN	O	O
of	NN	O	O
bacillus	NN	O	O
Calmette-Guerin	NN	O	O
(	NN	O	O
BCG	NN	O	O
)	NN	O	O
infection	NN	O	O
and	NN	O	O
are	NN	O	O
released	NN	O	O
by	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
upon	NN	O	O
stimulation	NN	O	O
by	NN	O	O
mycobacterial	NN	O	O
proteins	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
regard	NN	O	O
,	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
interleukin	NN	O	B-protein
6	NN	O	I-protein
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
and	NN	O	O
pathological	NN	O	O
events	NN	O	O
of	NN	O	O
tuberculosis	NN	O	O
infection	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
lipoarabinomannan	NN	O	B-protein
(	NN	O	O
LAM	NN	O	B-protein
)	NN	O	O
from	NN	O	O
the	NN	O	O
mycobacterial	NN	O	O
cell	NN	O	O
wall	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
virtually	NN	O	O
devoid	NN	O	O
of	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
stimulated	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
to	NN	O	O
release	NN	O	O
IL-6	NN	O	B-protein
in	NN	O	O
a	NN	O	O
dose-response	NN	O	O
manner	NN	O	O
.	NN	O	O

LAM	NN	O	B-protein
and	NN	O	O
LPS	NN	O	O
were	NN	O	O
potent	NN	O	O
inducers	NN	O	O
of	NN	O	O
IL-6	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Both	NN	O	O
LAM	NN	O	B-protein
-and	NN	O	O
LPS-inducible	NN	O	B-DNA
IL-6	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
was	NN	O	O
localized	NN	O	O
to	NN	O	O
a	NN	O	O
DNA	NN	O	B-DNA
fragment	NN	O	I-DNA
,	NN	O	O
positions	NN	O	O
-158	NN	O	B-DNA
to	NN	O	I-DNA
-49	NN	O	I-DNA
,	NN	O	O
by	NN	O	O
deletion	NN	O	O
analysis	NN	O	O
and	NN	O	O
chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
assay	NN	O	O
.	NN	O	O

Two	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
NF-IL6	NN	O	I-protein
(	NN	O	O
positions	NN	O	O
-153	NN	O	B-DNA
to	NN	O	I-DNA
-145	NN	O	I-DNA
and	NN	O	O
-83	NN	O	O
to	NN	O	O
-75	NN	O	O
)	NN	O	O
and	NN	O	O
one	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
(	NN	O	I-DNA
positions	NN	O	I-DNA
-72	NN	O	I-DNA
to	NN	O	I-DNA
-63	NN	O	I-DNA
)	NN	O	I-DNA
motifs	NN	O	I-DNA
are	NN	O	O
present	NN	O	O
within	NN	O	O
this	NN	O	O
fragment	NN	O	O
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
of	NN	O	O
these	NN	O	O
motifs	NN	O	O
within	NN	O	O
the	NN	O	O
IL-6	NN	O	B-protein
promoter	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
each	NN	O	O
has	NN	O	O
positive	NN	O	O
regulatory	NN	O	O
activity	NN	O	O
and	NN	O	O
that	NN	O	O
they	NN	O	O
could	NN	O	O
act	NN	O	O
in	NN	O	O
a	NN	O	O
function-	NN	O	O
and	NN	O	O
orientation-independent	NN	O	O
manner	NN	O	O
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
all	NN	O	O
three	NN	O	O
elements	NN	O	O
abolished	NN	O	O
inducibility	NN	O	O
of	NN	O	O
IL-6	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
by	NN	O	O
both	NN	O	O
LAM	NN	O	B-protein
and	NN	O	O
LPS	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
NF-IL6	NN	O	B-DNA
and	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
mediate	NN	O	O
IL-6	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
both	NN	O	O
LPS	NN	O	O
and	NN	O	O
LAM	NN	O	B-protein
,	NN	O	O
acting	NN	O	O
as	NN	O	O
bacterial	NN	O	B-DNA
or	NN	O	I-DNA
mycobacterial	NN	O	I-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
CD14	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
with	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
.	NN	O	O

CD14	NN	O	B-protein
,	NN	O	O
a	NN	O	O
monocyte/macrophage	NN	O	B-protein
receptor	NN	O	I-protein
for	NN	O	O
the	NN	O	O
complex	NN	O	O
of	NN	O	O
LPS	NN	O	O
and	NN	O	O
LPS	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
is	NN	O	O
a	NN	O	O
differentiation	NN	O	O
marker	NN	O	O
for	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
CD14	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
VitD3	NN	O	O
)	NN	O	O
-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Using	NN	O	O
FACS	NN	O	O
,	NN	O	O
Northern	NN	O	O
blotting	NN	O	O
,	NN	O	O
and	NN	O	O
nuclear	NN	O	O
run-on	NN	O	O
analyses	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
CD14	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
maturation	NN	O	O
is	NN	O	O
regulated	NN	O	O
mainly	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
new	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
CD14	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
cloned	NN	O	O
the	NN	O	O
CD14	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
upstream	NN	O	I-DNA
sequence	NN	O	I-DNA
and	NN	O	O
demonstrated	NN	O	O
its	NN	O	O
tissue-specific	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

Using	NN	O	O
stable	NN	O	O
transfection	NN	O	O
of	NN	O	O
the	NN	O	O
monocytoid	NN	O	B-cell_line
U937	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
with	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
deletion	NN	O	O
mutants	NN	O	O
of	NN	O	O
the	NN	O	O
CD14	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
upstream	NN	O	I-DNA
sequence	NN	O	I-DNA
coupled	NN	O	O
to	NN	O	O
a	NN	O	O
reporter	NN	O	O
gene	NN	O	O
construct	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
bp	NN	O	B-DNA
-128	NN	O	I-DNA
to	NN	O	I-DNA
-70	NN	O	I-DNA
is	NN	O	O
the	NN	O	O
critical	NN	O	O
region	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
CD14	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

This	NN	O	O
region	NN	O	O
contains	NN	O	O
two	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
Sp1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

A	NN	O	O
3-bp	NN	O	O
mutation	NN	O	O
at	NN	O	O
the	NN	O	O
distal	NN	O	O
Sp1-binding	NN	O	B-DNA
site	NN	O	I-DNA
not	NN	O	O
only	NN	O	O
eliminates	NN	O	O
Sp1	NN	O	B-protein
interaction	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
abolishes	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
VitD3	NN	O	O
induction	NN	O	O
of	NN	O	O
CD14	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analysis	NN	O	O
does	NN	O	O
not	NN	O	O
detect	NN	O	O
a	NN	O	O
direct	NN	O	O
interaction	NN	O	O
of	NN	O	O
the	NN	O	O
CD14	NN	O	B-protein
distal	NN	O	O
Sp1-binding	NN	O	B-DNA
site	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D3	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
its	NN	O	O
partner	NN	O	O
,	NN	O	O
the	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
VitD3	NN	O	O
induces	NN	O	O
CD14	NN	O	B-protein
indirectly	NN	O	O
through	NN	O	O
some	NN	O	O
intermediary	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
for	NN	O	O
Sp1	NN	O	B-protein
in	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

-DOCSTART-	O

DNA-binding	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
regulatory	NN	O	O
properties	NN	O	O
of	NN	O	O
hepatic	NN	O	B-protein
leukemia	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
HLF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
t	NN	O	O
(	NN	O	O
17	NN	O	O
;	NN	O	O
19	NN	O	O
)	NN	O	O
acute	NN	O	B-protein
lymphoblastic	NN	O	I-protein
leukemia	NN	O	I-protein
chimera	NN	O	I-protein
E2A-HLF	NN	O	I-protein
.	NN	O	O

The	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
17	NN	O	I-DNA
;	NN	O	I-DNA
19	NN	O	I-DNA
)	NN	O	I-DNA
translocation	NN	O	O
in	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemias	NN	O	O
results	NN	O	O
in	NN	O	O
creation	NN	O	O
of	NN	O	O
E2A-hepatic	NN	O	B-protein
leukemia	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
HLF	NN	O	B-protein
)	NN	O	O
chimeric	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
contain	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
and	NN	O	I-protein
protein	NN	O	I-protein
dimerization	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
leucine	NN	O	I-protein
zipper	NN	O	I-protein
(	NN	O	I-protein
bZIP	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
HLF	NN	O	B-protein
fused	NN	O	O
to	NN	O	O
a	NN	O	O
portion	NN	O	O
of	NN	O	O
E2A	NN	O	B-protein
proteins	NN	O	I-protein
with	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
properties	NN	O	O
.	NN	O	O

An	NN	O	O
in	NN	O	O
vitro	NN	O	O
binding	NN	O	O
site	NN	O	O
selection	NN	O	O
procedure	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
preferentially	NN	O	O
bound	NN	O	O
by	NN	O	O
wild-type	NN	O	B-protein
HLF	NN	O	I-protein
and	NN	O	O
chimeric	NN	O	B-protein
E2A-HLF	NN	O	I-protein
proteins	NN	O	O
isolated	NN	O	O
from	NN	O	O
various	NN	O	O
t	NN	O	B-cell_line
(	NN	O	I-cell_line
17	NN	O	I-cell_line
;	NN	O	I-cell_line
19	NN	O	I-cell_line
)	NN	O	I-cell_line
-bearing	NN	O	I-cell_line
leukemias	NN	O	I-cell_line
.	NN	O	O

All	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
selectively	NN	O	O
bind	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
sequence	NN	O	I-DNA
5'-GTTACGTAAT-3	NN	O	O
'	NN	O	O
with	NN	O	O
high	NN	O	O
affinity	NN	O	O
.	NN	O	O

Wild-type	NN	O	O
and	NN	O	O
chimeric	NN	O	B-protein
HLF	NN	O	I-protein
proteins	NN	O	I-protein
also	NN	O	O
bound	NN	O	O
closely	NN	O	O
related	NN	O	O
sites	NN	O	O
identified	NN	O	O
previously	NN	O	O
for	NN	O	O
bZIP	NN	O	B-protein
proteins	NN	O	I-protein
of	NN	O	O
both	NN	O	O
the	NN	O	O
proline	NN	O	B-protein
-and	NN	O	O
acidic	NN	O	O
amino	NN	O	O
acid-rich	NN	O	O
(	NN	O	O
PAR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
C/EBP	NN	O	B-protein
subfamilies	NN	O	I-protein
;	NN	O	O
however	NN	O	O
,	NN	O	O
E2A-HLF	NN	O	B-protein
proteins	NN	O	O
were	NN	O	O
significantly	NN	O	O
less	NN	O	O
tolerant	NN	O	O
of	NN	O	O
certain	NN	O	O
deviations	NN	O	O
from	NN	O	O
the	NN	O	O
HLF	NN	O	B-DNA
consensus	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
differences	NN	O	O
were	NN	O	O
directly	NN	O	O
attributable	NN	O	O
to	NN	O	O
loss	NN	O	O
of	NN	O	O
an	NN	O	O
HLF	NN	O	B-DNA
ancillary	NN	O	I-DNA
DNA-binding	NN	O	I-DNA
domain	NN	O	I-DNA
in	NN	O	O
all	NN	O	O
E2A-HLF	NN	O	B-protein
chimeras	NN	O	I-protein
and	NN	O	O
were	NN	O	O
further	NN	O	O
exacerbated	NN	O	O
by	NN	O	O
a	NN	O	O
zipper	NN	O	O
mutation	NN	O	O
in	NN	O	O
one	NN	O	O
isolate	NN	O	O
.	NN	O	O

Both	NN	O	O
wild-type	NN	O	B-protein
and	NN	O	O
chimeric	NN	O	B-protein
HLF	NN	O	I-protein
proteins	NN	O	I-protein
displayed	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
properties	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
and	NN	O	I-cell_type
nonlymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
on	NN	O	O
reporter	NN	O	B-DNA
genes	NN	O	I-DNA
containing	NN	O	O
HLF	NN	O	B-DNA
or	NN	O	I-DNA
C/EBP	NN	O	I-DNA
consensus	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

But	NN	O	O
on	NN	O	O
reporter	NN	O	B-DNA
genes	NN	O	I-DNA
with	NN	O	O
nonoptimal	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
their	NN	O	O
transcriptional	NN	O	O
properties	NN	O	O
diverged	NN	O	O
and	NN	O	O
E2A-HLF	NN	O	B-protein
competitively	NN	O	O
inhibited	NN	O	O
activation	NN	O	O
by	NN	O	O
wild-type	NN	O	B-protein
PAR	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
establish	NN	O	O
a	NN	O	O
spectrum	NN	O	O
of	NN	O	O
binding	NN	O	O
site-specific	NN	O	O
transcriptional	NN	O	O
properties	NN	O	O
for	NN	O	O
E2A-HLF	NN	O	B-protein
which	NN	O	O
may	NN	O	O
preferentially	NN	O	O
activate	NN	O	O
expression	NN	O	O
of	NN	O	O
select	NN	O	O
subordinate	NN	O	O
genes	NN	O	O
as	NN	O	O
a	NN	O	O
homodimer	NN	O	O
and	NN	O	O
potentially	NN	O	O
antagonize	NN	O	O
expression	NN	O	O
of	NN	O	O
others	NN	O	O
through	NN	O	O
heteromeric	NN	O	O
interactions	NN	O	O
.	NN	O	O

-DOCSTART-	O

ZAP-70	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
CD45	NN	O	B-protein
,	NN	O	O
and	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
involvement	NN	O	O
in	NN	O	O
UV-	NN	O	O
and	NN	O	O
H2O2-induced	NN	O	O
T	NN	O	O
cell	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

Several	NN	O	O
mammalian	NN	O	O
responses	NN	O	O
to	NN	O	O
UV	NN	O	O
irradiation	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
are	NN	O	O
believed	NN	O	O
to	NN	O	O
involve	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

UV	NN	O	O
irradiation	NN	O	O
and	NN	O	O
H2O2	NN	O	O
treatment	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
induce	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
signals	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
observed	NN	O	O
following	NN	O	O
biological	NN	O	O
stimulation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
cell	NN	O	O
surface	NN	O	O
molecules	NN	O	O
in	NN	O	O
these	NN	O	O
responses	NN	O	O
.	NN	O	O

Normal	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
whose	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
CD3	NN	O	B-protein
was	NN	O	O
depleted	NN	O	O
showed	NN	O	O
impaired	NN	O	O
UV-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
signals	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
deficient	NN	O	O
in	NN	O	O
CD3	NN	O	O
or	NN	O	O
CD45	NN	O	O
expression	NN	O	O
also	NN	O	O
gave	NN	O	O
impaired	NN	O	O
UV	NN	O	O
responses	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
all	NN	O	O
these	NN	O	O
cell	NN	O	O
types	NN	O	O
still	NN	O	O
gave	NN	O	O
strong	NN	O	O
Ca2+	NN	O	O
and	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
responses	NN	O	O
to	NN	O	O
H2O2	NN	O	O
.	NN	O	O

The	NN	O	O
T	NN	O	O
cell	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	B-protein
ZAP-70	NN	O	I-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
highly	NN	O	O
responsive	NN	O	O
to	NN	O	O
UV	NN	O	O
and	NN	O	O
H2O2	NN	O	O
treatment	NN	O	O
.	NN	O	O

ZAP-70	NN	O	B-protein
responsiveness	NN	O	O
to	NN	O	O
UV	NN	O	O
required	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
CD3	NN	O	B-protein
and	NN	O	O
CD45	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
only	NN	O	O
CD3	NN	O	B-protein
was	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
H2O2	NN	O	O
.	NN	O	O

UV-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
CD3	NN	O	B-protein
depletion	NN	O	O
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
such	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
biological	NN	O	O
responses	NN	O	O
to	NN	O	O
UV	NN	O	O
.	NN	O	O

In	NN	O	O
nonlymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
epidermal	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
displayed	NN	O	O
increased	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
within	NN	O	O
seconds	NN	O	O
of	NN	O	O
UV	NN	O	O
irradiation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
UV-induced	NN	O	O
signal	NN	O	O
transduction	NN	O	O
is	NN	O	O
mediated	NN	O	O
via	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
that	NN	O	O
normally	NN	O	O
respond	NN	O	O
to	NN	O	O
biological	NN	O	O
stimulation	NN	O	O
,	NN	O	O
whereas	NN	O	O
H2O2	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
partially	NN	O	O
bypass	NN	O	O
this	NN	O	O
requirement	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
by	NN	O	O
redox	NN	O	O
changes	NN	O	O
induced	NN	O	O
through	NN	O	O
glutathione	NN	O	O
depletion	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
redox	NN	O	O
changes	NN	O	O
in	NN	O	O
vivo	NN	O	O
on	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
two	NN	O	O
human	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
HL-60	NN	O	B-cell_line
and	NN	O	O
KG-1	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
glutathione-depleting	NN	O	O
agent	NN	O	O
diethyl	NN	O	O
maleate	NN	O	O
(	NN	O	O
DEM	NN	O	O
)	NN	O	O
prevented	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
differentiated	NN	O	O
features	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
,	NN	O	O
including	NN	O	O
adherence	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
to	NN	O	O
plastic	NN	O	O
surfaces	NN	O	O
and	NN	O	O
repression	NN	O	O
of	NN	O	O
the	NN	O	O
myeloperoxidase	NN	O	B-DNA
and	NN	O	I-DNA
CD34	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
DEM	NN	O	O
abolished	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP-1	NN	O	B-protein
and	NN	O	O
Egr-1	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
inhibition	NN	O	O
of	NN	O	O
differentiation	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
to	NN	O	O
redox	NN	O	O
modifications	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Lipopolysaccharide	NN	O	O
induction	NN	O	O
of	NN	O	O
tissue	NN	O	B-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
binding	NN	O	O
of	NN	O	O
c-Rel/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
to	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
B-like	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
activates	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
proteins	NN	O	O
and	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
rapid	NN	O	O
induction	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
,	NN	O	O
including	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
TF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

TF	NN	O	B-protein
is	NN	O	O
the	NN	O	O
primary	NN	O	O
cellular	NN	O	O
initiator	NN	O	O
of	NN	O	O
the	NN	O	O
coagulation	NN	O	B-protein
protease	NN	O	I-protein
cascades	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
complex	NN	O	I-protein
from	NN	O	O
human	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
bound	NN	O	O
to	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
B-like	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
5'-CGGAGTTTCC-3	NN	O	B-DNA
'	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
TF	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
nuclear	NN	O	B-protein
complex	NN	O	I-protein
was	NN	O	O
activated	NN	O	O
by	NN	O	O
LPS	NN	O	O
with	NN	O	O
kinetics	NN	O	O
that	NN	O	O
preceded	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
TF	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
binding	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
TF	NN	O	B-DNA
site	NN	O	I-DNA
bound	NN	O	O
translated	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	I-protein
p65	NN	O	I-protein
homodimers	NN	O	I-protein
but	NN	O	O
not	NN	O	O
p50/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
or	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	I-protein
.	NN	O	O

Base-pair	NN	O	O
substitutions	NN	O	O
in	NN	O	O
the	NN	O	O
TF	NN	O	B-DNA
site	NN	O	I-DNA
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
cytosine	NN	O	O
at	NN	O	O
position	NN	O	B-protein
1	NN	O	I-protein
precluded	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
under	NN	O	O
low-ionic-strength	NN	O	O
conditions	NN	O	O
,	NN	O	O
the	NN	O	O
TF	NN	O	B-protein
complex	NN	O	I-protein
did	NN	O	O
not	NN	O	O
migrate	NN	O	O
with	NN	O	O
translated	NN	O	O
p50/p65	NN	O	B-protein
dimers	NN	O	I-protein
but	NN	O	O
instead	NN	O	O
comigrated	NN	O	O
with	NN	O	O
c-Rel/p65	NN	O	B-protein
dimers	NN	O	I-protein
.	NN	O	O

Antibodies	NN	O	O
against	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
Rel	NN	O	O
proteins	NN	O	O
and	NN	O	O
UV	NN	O	O
cross-linking	NN	O	O
studies	NN	O	O
revealed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	O
p65	NN	O	O
and	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
in	NN	O	O
the	NN	O	O
TF	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
further	NN	O	O
showed	NN	O	O
that	NN	O	O
c-Rel/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
selectively	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
TF	NN	O	B-DNA
kappa	NN	O	I-DNA
B-like	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Functional	NN	O	O
studies	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
TF	NN	O	B-DNA
site	NN	O	I-DNA
conferred	NN	O	O
LPS	NN	O	O
inducibility	NN	O	O
on	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
was	NN	O	O
transactivated	NN	O	O
by	NN	O	O
c-Rel	NN	O	B-protein
or	NN	O	O
p65	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
c-Rel/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
to	NN	O	O
a	NN	O	O
novel	NN	O	O
kappa	NN	O	B-DNA
B-like	NN	O	I-DNA
site	NN	O	I-DNA
mediated	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
by	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
matrix	NN	O	I-protein
protein	NN	O	I-protein
tenascin	NN	O	B-protein
.	NN	O	O

Tenascin	NN	O	B-protein
(	NN	O	O
TN	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
extracellular	NN	O	B-protein
matrix	NN	O	I-protein
protein	NN	O	I-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
widely	NN	O	O
in	NN	O	O
the	NN	O	O
fetus	NN	O	O
and	NN	O	O
sparingly	NN	O	O
in	NN	O	O
the	NN	O	O
adult	NN	O	O
,	NN	O	O
but	NN	O	O
reappears	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
in	NN	O	O
certain	NN	O	O
areas	NN	O	O
of	NN	O	O
tissue	NN	O	O
insult	NN	O	O
such	NN	O	O
as	NN	O	O
tumor	NN	O	O
matrices	NN	O	O
and	NN	O	O
sites	NN	O	O
of	NN	O	O
wound	NN	O	O
healing	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
soluble	NN	O	O
TN	NN	O	B-protein
inhibits	NN	O	O
proliferation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
alpha	NN	O	B-protein
CD3	NN	O	I-protein
Ab	NN	O	I-protein
co-immobilized	NN	O	O
with	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
matrix	NN	O	I-protein
protein	NN	O	I-protein
fibronectin	NN	O	B-protein
(	NN	O	O
FN	NN	O	B-protein
)	NN	O	O
.	NN	O	O

TN	NN	O	B-protein
also	NN	O	O
inhibits	NN	O	O
proliferation	NN	O	O
driven	NN	O	O
by	NN	O	O
alpha	NN	O	B-protein
CD3/IL-2	NN	O	I-protein
or	NN	O	O
by	NN	O	O
phorbol	NN	O	B-protein
ester/IL-2	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	O
it	NN	O	O
prevents	NN	O	O
high	NN	O	O
level	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
TN	NN	O	B-protein
in	NN	O	O
culture	NN	O	O
medium	NN	O	O
does	NN	O	O
not	NN	O	O
detectably	NN	O	O
alter	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
resulting	NN	O	O
from	NN	O	O
T	NN	O	O
cell	NN	O	O
triggering	NN	O	O
with	NN	O	O
alpha	NN	O	B-protein
CD3	NN	O	I-protein
,	NN	O	O
but	NN	O	O
at	NN	O	O
later	NN	O	O
time	NN	O	O
points	NN	O	O
prevents	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
functional	NN	O	O
NF-AT1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
T	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
postulated	NN	O	O
role	NN	O	O
for	NN	O	O
TN	NN	O	B-protein
as	NN	O	O
a	NN	O	O
natural	NN	O	O
antagonist	NN	O	O
to	NN	O	O
FN	NN	O	B-protein
action	NN	O	O
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
occurring	NN	O	O
at	NN	O	O
tissue	NN	O	O
sites	NN	O	O
in	NN	O	O
which	NN	O	O
TN	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
could	NN	O	O
be	NN	O	O
influenced	NN	O	O
by	NN	O	O
its	NN	O	O
presence	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
Nef	NN	O	O
protein	NN	O	O
down-regulates	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
after	NN	O	O
T-cell	NN	O	O
receptor	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
negative	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
Nef	NN	O	B-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
down-regulate	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

To	NN	O	O
define	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
the	NN	O	O
Nef	NN	O	B-protein
protein	NN	O	O
,	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
affected	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
nef	NN	O	O
gene	NN	O	O
were	NN	O	O
examined	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor	NN	O	O
,	NN	O	O
interleukin-1	NN	O	O
,	NN	O	O
or	NN	O	O
lipopolysaccharide	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
factors	NN	O	O
to	NN	O	O
a	NN	O	O
similar	NN	O	O
level	NN	O	O
whether	NN	O	O
or	NN	O	O
not	NN	O	O
the	NN	O	O
cells	NN	O	O
expressed	NN	O	O
the	NN	O	O
nef	NN	O	O
gene	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
mitogens	NN	O	O
or	NN	O	O
antibodies	NN	O	O
to	NN	O	O
the	NN	O	O
T-cell	NN	O	O
receptor	NN	O	O
(	NN	O	O
TCR	NN	O	O
)	NN	O	O
-	NN	O	O
CD3	NN	O	B-protein
complex	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
factors	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
nef	NN	O	O
gene	NN	O	O
compared	NN	O	O
with	NN	O	O
cells	NN	O	O
not	NN	O	O
expressing	NN	O	O
the	NN	O	O
nef	NN	O	O
gene	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
Nef	NN	O	B-protein
protein	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
-TCR	NN	O	O
complex	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
Nef	NN	O	B-protein
protein	NN	O	O
down-regulates	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
factors	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
through	NN	O	O
an	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
TCR-dependent	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
alpha-lipoic	NN	O	O
acid	NN	O	O
and	NN	O	O
dihydrolipoic	NN	O	O
acid	NN	O	O
on	NN	O	O
expression	NN	O	O
of	NN	O	O
proto-oncogene	NN	O	B-DNA
c-fos	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
human	NN	O	O
mediator	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	O
response	NN	O	O
to	NN	O	O
serum	NN	O	O
,	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
such	NN	O	O
as	NN	O	O
12-O-tetradecanoyl-phorbol-13	NN	O	O
acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
AP-1	NN	O	B-protein
complex	NN	O	O
consists	NN	O	O
of	NN	O	O
distinct	NN	O	O
protein	NN	O	B-protein
heterodimers	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
proto-oncogene	NN	O	B-DNA
c-fos	NN	O	I-DNA
and	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
whose	NN	O	O
gene	NN	O	O
expression	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
TPA	NN	O	O
,	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
and	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Recent	NN	O	O
findings	NN	O	O
suggest	NN	O	O
an	NN	O	O
involvement	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
in	NN	O	O
the	NN	O	O
pathway	NN	O	O
of	NN	O	O
TPA	NN	O	O
and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
leading	NN	O	O
to	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
and	NN	O	I-RNA
c-jun	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
alpha-lipoic	NN	O	O
acid	NN	O	O
and	NN	O	O
dihydrolipoic	NN	O	O
acid	NN	O	O
(	NN	O	O
natural	NN	O	O
thiol	NN	O	O
antioxidants	NN	O	O
)	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

When	NN	O	O
cells	NN	O	O
were	NN	O	O
preincubated	NN	O	O
with	NN	O	O
dihydrolipoic	NN	O	O
acid	NN	O	O
(	NN	O	O
0.2	NN	O	O
mM	NN	O	O
)	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
suppressed	NN	O	O
at	NN	O	O
30	NN	O	O
min	NN	O	O
after	NN	O	O
stimulation	NN	O	O
of	NN	O	O
TPA	NN	O	O
(	NN	O	O
0.5	NN	O	O
microM	NN	O	O
)	NN	O	O
whereas	NN	O	O
in	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
preincubation	NN	O	O
of	NN	O	O
alpha-lipoic	NN	O	O
acid	NN	O	O
(	NN	O	O
0.2	NN	O	O
microM	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
was	NN	O	O
enhanced	NN	O	O
at	NN	O	O
30	NN	O	O
min	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
support	NN	O	O
the	NN	O	O
idea	NN	O	O
that	NN	O	O
superoxide	NN	O	O
anion	NN	O	O
radical	NN	O	O
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

-DOCSTART-	O

Appraisal	NN	O	O
of	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
index	NN	O	O
of	NN	O	O
antioxidants	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
their	NN	O	O
in	NN	O	O
vitro	NN	O	O
effects	NN	O	O
on	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
-induced	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Antioxidant	NN	O	O
molecules	NN	O	O
have	NN	O	O
been	NN	O	O
suggested	NN	O	O
to	NN	O	O
be	NN	O	O
of	NN	O	O
therapeutic	NN	O	O
value	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
HIV-infected	NN	O	O
patients	NN	O	O
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
this	NN	O	O
possibility	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
in	NN	O	O
vitro	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
two	NN	O	O
types	NN	O	O
of	NN	O	O
antioxidant	NN	O	O
molecules	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
inhibition	NN	O	O
of	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
main	NN	O	O
reservoirs	NN	O	O
of	NN	O	O
HIV	NN	O	O
,	NN	O	O
and	NN	O	O
also	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
competence	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
PBMC	NN	O	B-cell_type
proliferation	NN	O	O
.	NN	O	O

We	NN	O	O
tested	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
BHA	NN	O	O
,	NN	O	O
a	NN	O	O
phenolic	NN	O	O
,	NN	O	O
lipid-soluble	NN	O	O
,	NN	O	O
chain-breaking	NN	O	O
antioxidant	NN	O	O
,	NN	O	O
and	NN	O	O
NAC	NN	O	O
,	NN	O	O
a	NN	O	O
known	NN	O	O
glutathione	NN	O	O
precursor	NN	O	O
with	NN	O	O
some	NN	O	O
direct	NN	O	O
free-radical	NN	O	O
scavenging	NN	O	O
properties	NN	O	O
as	NN	O	O
well	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
in	NN	O	O
latently	NN	O	O
infected	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
productively	NN	O	O
and	NN	O	O
chronically	NN	O	O
infected	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Both	NN	O	O
antioxidants	NN	O	O
inhibited	NN	O	O
TNF	NN	O	B-protein
-or	NN	O	O
PMA-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
sustained	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
permanently	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
virus	NN	O	O
itself	NN	O	O
in	NN	O	O
chronically	NN	O	O
HIV-infected	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
resulted	NN	O	O
in	NN	O	O
only	NN	O	O
a	NN	O	O
partial	NN	O	O
inhibition	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
-or	NN	O	O
PMA-	NN	O	O
induced	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
no	NN	O	O
detectable	NN	O	O
effect	NN	O	O
on	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
chronically	NN	O	B-cell_line
infected	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
may	NN	O	O
be	NN	O	O
the	NN	O	O
first	NN	O	O
limitation	NN	O	O
to	NN	O	O
potential	NN	O	O
antiviral	NN	O	O
effects	NN	O	O
of	NN	O	O
antioxidant	NN	O	O
therapies	NN	O	O
.	NN	O	O

Another	NN	O	O
limitation	NN	O	O
is	NN	O	O
that	NN	O	O
antioxidant	NN	O	O
concentrations	NN	O	O
high	NN	O	O
enough	NN	O	O
to	NN	O	O
block	NN	O	O
NK-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
suppressive	NN	O	O
effect	NN	O	O
on	NN	O	O
immune	NN	O	O
functions	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
because	NN	O	O
NAC	NN	O	O
and	NN	O	O
BHA	NN	O	O
blocked	NN	O	O
IL-2-induced	NN	O	B-cell_type
PBMC	NN	O	I-cell_type
proliferation	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
warrant	NN	O	O
prudence	NN	O	O
in	NN	O	O
the	NN	O	O
design	NN	O	O
of	NN	O	O
antioxidant-based	NN	O	O
therapies	NN	O	O
aimed	NN	O	O
at	NN	O	O
suppressing	NN	O	O
HIV	NN	O	O
replication	NN	O	O
.	NN	O	O

-DOCSTART-	O

Isolation	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
gelatinase	NN	O	O
granules	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
recently	NN	O	O
confirmed	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
gelatinase	NN	O	O
granules	NN	O	O
as	NN	O	O
a	NN	O	O
subpopulation	NN	O	O
of	NN	O	O
peroxidase-negative	NN	O	O
granules	NN	O	O
by	NN	O	O
double-labeling	NN	O	O
immunogold	NN	O	O
electron	NN	O	O
microscopy	NN	O	O
on	NN	O	O
intact	NN	O	O
cells	NN	O	O
and	NN	O	O
by	NN	O	O
subcellular	NN	O	O
fractionation	NN	O	O
.	NN	O	O

Further	NN	O	O
characterization	NN	O	O
of	NN	O	O
gelatinase	NN	O	O
granules	NN	O	O
has	NN	O	O
been	NN	O	O
hampered	NN	O	O
by	NN	O	O
poor	NN	O	O
separation	NN	O	O
of	NN	O	O
specific	NN	O	O
and	NN	O	O
gelatinase	NN	O	O
granules	NN	O	O
on	NN	O	O
both	NN	O	O
two-layer	NN	O	O
Percoll	NN	O	O
gradients	NN	O	O
and	NN	O	O
sucrose	NN	O	O
gradients	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
three-layer	NN	O	O
Percoll	NN	O	O
density	NN	O	O
gradient	NN	O	O
that	NN	O	O
allows	NN	O	O
separation	NN	O	O
of	NN	O	O
the	NN	O	O
different	NN	O	O
granules	NN	O	O
and	NN	O	O
vesicles	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
;	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
it	NN	O	O
allows	NN	O	O
separation	NN	O	O
of	NN	O	O
specific	NN	O	O
and	NN	O	O
gelatinase	NN	O	O
granules	NN	O	O
.	NN	O	O

This	NN	O	O
allows	NN	O	O
us	NN	O	O
to	NN	O	O
characterize	NN	O	O
these	NN	O	O
two	NN	O	O
granule	NN	O	O
populations	NN	O	O
with	NN	O	O
regard	NN	O	O
to	NN	O	O
their	NN	O	O
content	NN	O	O
of	NN	O	O
membrane	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
which	NN	O	O
become	NN	O	O
incorporated	NN	O	O
into	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
during	NN	O	O
exocytosis	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
gelatinase	NN	O	O
granules	NN	O	O
,	NN	O	O
defined	NN	O	O
as	NN	O	O
peroxidase-negative	NN	O	O
granules	NN	O	O
containing	NN	O	O
gelatinase	NN	O	B-protein
but	NN	O	O
lacking	NN	O	O
lactoferrin	NN	O	B-protein
,	NN	O	O
contain	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
total	NN	O	O
cell	NN	O	O
gelatinase	NN	O	B-protein
,	NN	O	O
with	NN	O	O
the	NN	O	O
remaining	NN	O	O
residing	NN	O	O
in	NN	O	O
specific	NN	O	O
granules	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
20	NN	O	O
%	NN	O	O
to	NN	O	O
25	NN	O	O
%	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
adhesion	NN	O	B-protein
protein	NN	O	I-protein
Mac-1	NN	O	I-protein
and	NN	O	O
the	NN	O	O
NADPH-oxidase	NN	O	B-protein
component	NN	O	I-protein
cytochrome	NN	O	I-protein
b558	NN	O	B-protein
is	NN	O	O
localized	NN	O	O
in	NN	O	O
gelatinase	NN	O	B-protein
granules	NN	O	O
.	NN	O	O

Although	NN	O	O
no	NN	O	O
qualitative	NN	O	O
difference	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
specific	NN	O	O
granules	NN	O	O
and	NN	O	O
gelatinase	NN	O	B-protein
granules	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
cytochrome	NN	O	B-protein
b558	NN	O	I-protein
and	NN	O	O
Mac-1	NN	O	B-protein
,	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
neutrophil	NN	O	B-cell_type
with	NN	O	O
FMLP	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
selective	NN	O	O
mobilization	NN	O	O
of	NN	O	O
the	NN	O	O
least	NN	O	O
dense	NN	O	O
peroxidase-negative	NN	O	O
granules	NN	O	O
,	NN	O	O
ie	NN	O	O
,	NN	O	O
gelatinase	NN	O	B-protein
granules	NN	O	O
,	NN	O	O
which	NN	O	O
,	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
secretory	NN	O	O
vesicles	NN	O	O
,	NN	O	O
furnish	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
with	NN	O	O
Mac-1	NN	O	B-protein
and	NN	O	O
cytochrome	NN	O	B-protein
b558	NN	O	I-protein
.	NN	O	O

This	NN	O	O
shows	NN	O	O
that	NN	O	O
gelatinase	NN	O	B-protein
granules	NN	O	O
are	NN	O	O
functionally	NN	O	O
important	NN	O	O
relative	NN	O	O
to	NN	O	O
specific	NN	O	O
granules	NN	O	O
in	NN	O	O
mediating	NN	O	O
early	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor.kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Autocrine	NN	O	O
role	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
expression	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
NF	NN	O	O
)	NN	O	O
activation	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
has	NN	O	O
been	NN	O	O
studied	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
human	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
strongly	NN	O	O
induced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF.kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

This	NN	O	O
activation	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
translocation	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
NF.kappa	NN	O	B-protein
B	NN	O	I-protein
proteins	NN	O	O
from	NN	O	O
cytoplasmic	NN	O	O
stores	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
where	NN	O	O
they	NN	O	O
bound	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
either	NN	O	O
as	NN	O	O
p50.p65	NN	O	B-protein
or	NN	O	O
as	NN	O	O
p50.c-Rel	NN	O	B-protein
heterodimers	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
all	NN	O	O
of	NN	O	O
those	NN	O	O
events	NN	O	O
were	NN	O	O
largely	NN	O	O
indirect	NN	O	O
and	NN	O	O
mediated	NN	O	O
by	NN	O	O
endogenously	NN	O	O
secreted	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
,	NN	O	O
as	NN	O	O
they	NN	O	O
were	NN	O	O
strongly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
neutralizing	NN	O	B-protein
anti-TNF	NN	O	I-protein
alpha	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
which	NN	O	O
blocked	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
production	NN	O	O
induced	NN	O	O
by	NN	O	O
PKC	NN	O	B-protein
,	NN	O	O
strongly	NN	O	O
inhibited	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
NF.kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
either	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
or	NN	O	O
IL-2	NN	O	B-protein
partially	NN	O	O
recovered	NN	O	O
cyclosporin	NN	O	O
A-induced	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
inhibition	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
completely	NN	O	O
recovered	NN	O	O
NF.kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Those	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
PKC	NN	O	B-protein
activation	NN	O	O
has	NN	O	O
only	NN	O	O
a	NN	O	O
triggering	NN	O	O
role	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
endogenously	NN	O	O
secreted	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
plays	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
quantitative	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	O
or	NN	O	O
NF.kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Superantigens	NN	O	B-protein
activate	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
superantigens	NN	O	O
to	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
results	NN	O	O
in	NN	O	O
transduction	NN	O	O
of	NN	O	O
biochemical	NN	O	O
signals	NN	O	O
leading	NN	O	O
to	NN	O	O
cellular	NN	O	O
activation	NN	O	O
and	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
staphylococcal	NN	O	B-protein
superantigens	NN	O	I-protein
toxic	NN	O	B-protein
shock	NN	O	I-protein
syndrome	NN	O	I-protein
toxin-1	NN	O	I-protein
(	NN	O	O
TSST-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
staphylococcal	NN	O	B-protein
enterotoxin	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	O
SEA	NN	O	B-protein
)	NN	O	O
activate	NN	O	O
HIV-1-LTR	NN	O	B-DNA
-driven	NN	O	O
transcription	NN	O	O
of	NN	O	O
chloramphenicol	NN	O	B-protein
acetyl	NN	O	I-protein
transferase	NN	O	I-protein
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	B-cell_line
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
HIV-1-	NN	O	O
LTR	NN	O	B-DNA
-driven	NN	O	O
transcription	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
superantigens	NN	O	B-protein
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
DNA-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Superantigens	NN	O	B-protein
also	NN	O	O
increased	NN	O	O
viral	NN	O	O
protein	NN	O	O
secretion	NN	O	O
from	NN	O	O
the	NN	O	O
granulocyte-macrophage	NN	O	B-cell_line
colony-stimulating	NN	O	I-cell_line
factor-pretreated	NN	O	I-cell_line
chronically	NN	O	I-cell_line
infected	NN	O	I-cell_line
human	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U1	NN	O	I-cell_line
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
superantigens	NN	O	B-protein
occurred	NN	O	O
via	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor-alpha-dependent	NN	O	O
and	NN	O	O
-independent	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
superantigens	NN	O	B-protein
and	NN	O	O
other	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
ligands	NN	O	I-protein
may	NN	O	O
activate	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Mitogen	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
induces	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
new	NN	O	O
cyclic	NN	O	B-protein
AMP	NN	O	I-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
nuclear	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

A	NN	O	O
large	NN	O	O
body	NN	O	O
of	NN	O	O
evidence	NN	O	O
indicates	NN	O	O
that	NN	O	O
experimental	NN	O	O
agents	NN	O	O
which	NN	O	O
raise	NN	O	O
cellular	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
inhibit	NN	O	O
T	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
division	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
many	NN	O	O
studies	NN	O	O
have	NN	O	O
reported	NN	O	O
that	NN	O	O
mitogen	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
increases	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
,	NN	O	O
implying	NN	O	O
a	NN	O	O
positive	NN	O	O
physiological	NN	O	O
role	NN	O	O
for	NN	O	O
cAMP	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
process	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
mitogen	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
induces	NN	O	O
nuclear	NN	O	O
factors	NN	O	O
that	NN	O	O
form	NN	O	O
complexes	NN	O	O
with	NN	O	O
cyclic	NN	O	B-protein
AMP	NN	O	I-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Four	NN	O	O
complexes	NN	O	O
are	NN	O	O
identified	NN	O	O
by	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
two	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
induced	NN	O	O
by	NN	O	O
mitogen	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

All	NN	O	O
four	NN	O	O
complexes	NN	O	O
contain	NN	O	O
CREB	NN	O	B-protein
and	NN	O	O
are	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
cAMP	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
CRE	NN	O	I-DNA
)	NN	O	I-DNA
core	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
TGACGTCA	NN	O	O
)	NN	O	O
,	NN	O	O
as	NN	O	O
indicated	NN	O	O
by	NN	O	O
antibody	NN	O	O
and	NN	O	O
oligonucleotide	NN	O	O
competition	NN	O	O
experiments	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
the	NN	O	O
four	NN	O	O
complexes	NN	O	O
to	NN	O	O
CRE	NN	O	B-DNA
is	NN	O	O
prevented	NN	O	O
by	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
and	NN	O	O
is	NN	O	O
restored	NN	O	O
by	NN	O	O
rephosphorylation	NN	O	O
with	NN	O	O
cAMP-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
or	NN	O	O
endogenous	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

Similar	NN	O	O
complexes	NN	O	O
are	NN	O	O
detected	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Mitogen	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
electrophoretic	NN	O	B-protein
mobility	NN	O	I-protein
shift	NN	O	I-protein
assay	NN	O	I-protein
complexes	NN	O	I-protein
is	NN	O	O
not	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
protein	NN	O	O
phosphorylation	NN	O	O
or	NN	O	O
by	NN	O	O
induction	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
the	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
mitogen	NN	O	O
increases	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
a	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
that	NN	O	O
dimerizes	NN	O	O
with	NN	O	O
CREB	NN	O	B-protein
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
new	NN	O	O
CREB	NN	O	B-protein
complexes	NN	O	I-protein
implies	NN	O	O
a	NN	O	O
physiological	NN	O	O
role	NN	O	O
for	NN	O	O
cAMP	NN	O	O
in	NN	O	O
mitogen	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Alpha-tocopherol	NN	O	O
inhibits	NN	O	O
agonist-induced	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
to	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Antioxidants	NN	O	O
have	NN	O	O
been	NN	O	O
proposed	NN	O	O
to	NN	O	O
be	NN	O	O
anti-atherosclerotic	NN	O	O
agents	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
their	NN	O	O
beneficial	NN	O	O
effects	NN	O	O
are	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
alpha-tocopherol	NN	O	O
(	NN	O	O
alpha-tcp	NN	O	O
)	NN	O	O
on	NN	O	O
one	NN	O	O
cellular	NN	O	O
event	NN	O	O
in	NN	O	O
atherosclerotic	NN	O	O
plaque	NN	O	O
development	NN	O	O
,	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
stimulated	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
ECs	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

Human	NN	O	O
umbilical	NN	O	O
vein	NN	O	O
ECs	NN	O	B-cell_type
were	NN	O	O
pretreated	NN	O	O
with	NN	O	O
alpha-tcp	NN	O	O
before	NN	O	O
stimulation	NN	O	O
with	NN	O	O
known	NN	O	O
agonists	NN	O	O
of	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
:	NN	O	O
IL-1	NN	O	B-protein
(	NN	O	O
10	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
,	NN	O	O
LPS	NN	O	O
(	NN	O	O
10	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
,	NN	O	O
thrombin	NN	O	B-protein
(	NN	O	O
30	NN	O	O
U/ml	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
PMA	NN	O	O
(	NN	O	O
10	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

Agonist-induced	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
basal	NN	O	O
adhesion	NN	O	O
,	NN	O	O
was	NN	O	O
inhibited	NN	O	O
in	NN	O	O
a	NN	O	O
time-	NN	O	O
and	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
by	NN	O	O
alpha-tcp	NN	O	O
.	NN	O	O

The	NN	O	O
IC50	NN	O	O
of	NN	O	O
alpha-tcp	NN	O	O
on	NN	O	O
an	NN	O	O
IL-1	NN	O	B-protein
-induced	NN	O	O
response	NN	O	O
was	NN	O	O
45	NN	O	O
microM	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
steady	NN	O	O
state	NN	O	O
levels	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-protein
which	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
to	NN	O	O
E-selectin	NN	O	B-protein
to	NN	O	O
inhibit	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
in	NN	O	O
this	NN	O	O
system	NN	O	O
.	NN	O	O

Probucol	NN	O	O
(	NN	O	O
50	NN	O	O
microM	NN	O	O
)	NN	O	O
and	NN	O	O
N-acetylcysteine	NN	O	O
(	NN	O	O
20	NN	O	O
mM	NN	O	O
)	NN	O	O
also	NN	O	O
inhibited	NN	O	O
agonist-induced	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
;	NN	O	O
whereas	NN	O	O
,	NN	O	O
several	NN	O	O
other	NN	O	O
antioxidants	NN	O	O
had	NN	O	O
no	NN	O	O
significant	NN	O	O
effect	NN	O	O
.	NN	O	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
alpha-tcp	NN	O	O
effect	NN	O	O
since	NN	O	O
no	NN	O	O
suppression	NN	O	O
of	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
substrates	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
necessary	NN	O	O
but	NN	O	O
not	NN	O	O
sufficient	NN	O	O
for	NN	O	O
E-selectin	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
EC	NN	O	B-cell_line
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
failed	NN	O	O
to	NN	O	O
show	NN	O	O
an	NN	O	O
alpha-tcp-induced	NN	O	O
decrease	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
after	NN	O	O
cytokine	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
alpha-tcp	NN	O	O
acts	NN	O	O
as	NN	O	O
an	NN	O	O
anti-atherosclerotic	NN	O	O
molecule	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
generation	NN	O	O
of	NN	O	O
oxidized	NN	O	O
LDL	NN	O	O
--	NN	O	O
a	NN	O	O
putative	NN	O	O
triggering	NN	O	O
molecule	NN	O	O
in	NN	O	O
the	NN	O	O
atherosclerotic	NN	O	O
process	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
point	NN	O	O
to	NN	O	O
a	NN	O	O
novel	NN	O	O
alternative	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
alpha-tcp	NN	O	O
.	NN	O	O

-DOCSTART-	O

Central	NN	O	B-cell_type
nervous	NN	O	I-cell_type
system-derived	NN	O	I-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
a	NN	O	O
kappa	NN	O	O
B-binding	NN	O	O
activity	NN	O	O
that	NN	O	O
enhances	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
transcription	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
facilitates	NN	O	O
TAR	NN	O	B-DNA
-independent	NN	O	O
transactivation	NN	O	O
by	NN	O	O
Tat	NN	O	B-protein
.	NN	O	O

The	NN	O	O
Tat	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
activator	NN	O	O
of	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
-directed	NN	O	O
transcription	NN	O	O
.	NN	O	O

While	NN	O	O
in	NN	O	O
most	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
activation	NN	O	O
requires	NN	O	O
interaction	NN	O	O
of	NN	O	O
Tat	NN	O	O
with	NN	O	O
the	NN	O	O
unusual	NN	O	O
transcription	NN	O	B-DNA
element	NN	O	I-DNA
TAR	NN	O	B-DNA
,	NN	O	O
astrocytic	NN	O	B-cell_type
glial	NN	O	I-cell_type
cells	NN	O	I-cell_type
support	NN	O	O
TAR	NN	O	B-DNA
-independent	NN	O	O
transactivation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
by	NN	O	O
Tat	NN	O	O
.	NN	O	O

This	NN	O	O
alternative	NN	O	O
pathway	NN	O	O
of	NN	O	O
Tat	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
kappa	NN	O	O
B	NN	O	O
domain	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
the	NN	O	O
prototypical	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	O
factor	NN	O	O
kappa	NN	O	O
B	NN	O	O
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
present	NN	O	O
in	NN	O	O
many	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
including	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Tat	NN	O	B-protein
transactivation	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
domain	NN	O	I-DNA
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
allow	NN	O	O
replication	NN	O	O
of	NN	O	O
TAR	NN	O	B-DNA
-deleted	NN	O	O
mutant	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
astrocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
kappa	NN	O	B-protein
B-specific	NN	O	I-protein
binding	NN	O	I-protein
factors	NN	O	I-protein
present	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
glial	NN	O	I-cell_type
astrocytes	NN	O	I-cell_type
that	NN	O	O
differ	NN	O	O
from	NN	O	O
prototypical	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
novel	NN	O	O
astrocyte-derived	NN	O	O
kappa	NN	O	O
B-binding	NN	O	O
activity	NN	O	O
is	NN	O	O
retained	NN	O	O
on	NN	O	O
an	NN	O	O
HIV-1	NN	O	O
Tat	NN	O	B-protein
affinity	NN	O	O
column	NN	O	O
,	NN	O	O
while	NN	O	O
prototypical	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
from	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
not	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
astrocyte-derived	NN	O	B-protein
kappa	NN	O	I-protein
B-binding	NN	O	I-protein
factors	NN	O	I-protein
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
this	NN	O	O
activation	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
domain	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
TAR	NN	O	B-DNA
-independent	NN	O	O
transactivation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
is	NN	O	O
reproduced	NN	O	O
in	NN	O	O
vitro	NN	O	O
in	NN	O	O
an	NN	O	O
astrocyte	NN	O	O
factor-dependent	NN	O	O
manner	NN	O	O
which	NN	O	O
correlates	NN	O	O
with	NN	O	O
kappa	NN	O	O
B-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
importance	NN	O	O
of	NN	O	O
the	NN	O	O
central	NN	O	B-protein
nervous	NN	O	I-protein
system-enriched	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-protein
interleukin-13	NN	O	I-protein
activates	NN	O	O
the	NN	O	O
interleukin-4-dependent	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-IL4	NN	O	B-protein
sharing	NN	O	O
a	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
motif	NN	O	I-protein
with	NN	O	O
an	NN	O	O
interferon-gamma-induced	NN	O	B-protein
nuclear	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
interleukin-13	NN	O	B-protein
(	NN	O	O
IL-13	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
on	NN	O	O
cellular	NN	O	O
functions	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
quite	NN	O	O
similar	NN	O	O
.	NN	O	O

We	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
in	NN	O	O
monocytes	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
both	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
activate	NN	O	O
the	NN	O	O
same	NN	O	O
recently	NN	O	O
identified	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-IL4	NN	O	B-protein
which	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
specific	NN	O	B-DNA
responsive	NN	O	I-DNA
element	NN	O	I-DNA
IL-4RE	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
activated	NN	O	O
by	NN	O	O
interferon-gamma	NN	O	O
also	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
IL-4RE	NN	O	B-DNA
.	NN	O	O

It	NN	O	O
differs	NN	O	O
from	NN	O	O
NF-IL4	NN	O	B-protein
in	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
of	NN	O	O
the	NN	O	O
complex	NN	O	O
with	NN	O	O
DNA	NN	O	O
,	NN	O	O
in	NN	O	O
its	NN	O	O
DNA-binding	NN	O	O
specificity	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
proteins	NN	O	O
interacting	NN	O	O
with	NN	O	O
the	NN	O	O
DNA	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Sensitivity	NN	O	O
against	NN	O	O
various	NN	O	O
enzyme	NN	O	O
inhibitors	NN	O	O
suggests	NN	O	O
that	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
are	NN	O	O
shared	NN	O	O
by	NN	O	O
all	NN	O	O
three	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Encephalomyocarditis	NN	O	O
virus	NN	O	O
internal	NN	O	O
ribosomal	NN	O	O
entry	NN	O	O
site	NN	O	O
RNA-protein	NN	O	O
interactions	NN	O	O
.	NN	O	O

Translational	NN	O	O
initiation	NN	O	O
of	NN	O	O
encephalomyocarditis	NN	O	B-RNA
virus	NN	O	I-RNA
(	NN	O	I-RNA
EMCV	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
occurs	NN	O	O
by	NN	O	O
ribosomal	NN	O	O
entry	NN	O	O
into	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
nontranslated	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
EMCV	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
rather	NN	O	O
than	NN	O	O
by	NN	O	O
ribosomal	NN	O	O
scanning	NN	O	O
.	NN	O	O

Internal	NN	O	O
ribosomal	NN	O	O
binding	NN	O	O
requires	NN	O	O
a	NN	O	O
cis-acting	NN	O	B-DNA
element	NN	O	I-DNA
termed	NN	O	O
the	NN	O	O
internal	NN	O	B-DNA
ribosomal	NN	O	I-DNA
entry	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
IRES	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

IRES	NN	O	B-DNA
elements	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
proposed	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
translation	NN	O	O
of	NN	O	O
picornavirus	NN	O	B-RNA
mRNAs	NN	O	I-RNA
and	NN	O	O
some	NN	O	O
cellular	NN	O	B-RNA
mRNAs	NN	O	I-RNA
.	NN	O	O

Internal	NN	O	O
ribosome	NN	O	O
binding	NN	O	O
likely	NN	O	O
requires	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
trans-acting	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
recognize	NN	O	O
both	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
and	NN	O	O
the	NN	O	O
ribosomal	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

Five	NN	O	O
cellular	NN	O	B-protein
proteins	NN	O	I-protein
(	NN	O	O
p52	NN	O	B-protein
,	NN	O	O
p57	NN	O	B-protein
,	NN	O	O
p70	NN	O	B-protein
,	NN	O	O
p72	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p100	NN	O	B-protein
)	NN	O	O
cross-link	NN	O	O
the	NN	O	O
EMCV	NN	O	O
IRES	NN	O	B-DNA
or	NN	O	O
fragments	NN	O	O
of	NN	O	O
the	NN	O	O
IRES	NN	O	B-DNA
.	NN	O	O

For	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
,	NN	O	O
p57	NN	O	B-protein
,	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
IRES	NN	O	B-DNA
correlates	NN	O	O
with	NN	O	O
translation	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
p57	NN	O	B-protein
was	NN	O	O
identified	NN	O	O
to	NN	O	O
be	NN	O	O
very	NN	O	O
similar	NN	O	O
,	NN	O	O
if	NN	O	O
not	NN	O	O
identical	NN	O	O
,	NN	O	O
to	NN	O	O
polypyrimidine	NN	O	B-protein
tract-binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
cross-linking	NN	O	O
results	NN	O	O
with	NN	O	O
21	NN	O	O
different	NN	O	O
EMCV	NN	O	B-DNA
IRES	NN	O	I-DNA
fragments	NN	O	I-DNA
and	NN	O	O
cytoplasmic	NN	O	O
HeLa	NN	O	O
extract	NN	O	O
or	NN	O	O
rabbit	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
as	NN	O	O
the	NN	O	O
source	NN	O	O
of	NN	O	O
polypeptides	NN	O	O
,	NN	O	O
consensus	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
p52	NN	O	B-protein
,	NN	O	O
p57	NN	O	B-protein
,	NN	O	O
p70	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p100	NN	O	B-protein
are	NN	O	O
proposed	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
suggested	NN	O	O
that	NN	O	O
each	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
recognizes	NN	O	O
primarily	NN	O	O
a	NN	O	O
structural	NN	O	O
feature	NN	O	O
of	NN	O	O
the	NN	O	O
RNA	NN	O	O
rather	NN	O	O
than	NN	O	O
a	NN	O	O
specific	NN	O	O
sequence	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
heterodimerization	NN	O	B-protein
partner	NN	O	I-protein
for	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Retinoid-like	NN	O	B-protein
receptors	NN	O	I-protein
play	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
hormonal	NN	O	O
responses	NN	O	O
by	NN	O	O
forming	NN	O	O
heterodimers	NN	O	B-protein
with	NN	O	O
other	NN	O	O
nuclear	NN	O	B-protein
hormone	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
the	NN	O	O
peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
PPAR	NN	O	B-protein
)	NN	O	O
as	NN	O	O
a	NN	O	O
new	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
THR	NN	O	I-protein
)	NN	O	I-protein
auxiliary	NN	O	I-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
heterodimerizing	NN	O	O
with	NN	O	O
THR	NN	O	B-protein
in	NN	O	O
solution	NN	O	O
.	NN	O	O

Although	NN	O	O
these	NN	O	O
heterodimers	NN	O	B-protein
do	NN	O	O
not	NN	O	O
recognize	NN	O	O
a	NN	O	O
classical	NN	O	O
thyroid	NN	O	B-DNA
hormone	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
TRE	NN	O	B-DNA
)	NN	O	O
characterized	NN	O	O
by	NN	O	O
direct	NN	O	O
repeat	NN	O	O
separated	NN	O	O
by	NN	O	O
four	NN	O	O
nucleotides	NN	O	O
(	NN	O	O
DR+4	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
PPAR	NN	O	B-protein
behaves	NN	O	O
as	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
thyroid	NN	O	O
hormone	NN	O	O
(	NN	O	O
TH	NN	O	O
)	NN	O	O
action	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
TH-dependent	NN	O	O
positive	NN	O	O
effect	NN	O	O
is	NN	O	O
elicited	NN	O	O
by	NN	O	O
selective	NN	O	O
interaction	NN	O	O
of	NN	O	O
the	NN	O	O
THR	NN	O	B-protein
beta-PPAR	NN	O	I-protein
but	NN	O	O
not	NN	O	O
the	NN	O	O
THR	NN	O	B-protein
alpha-PPAR	NN	O	I-protein
heterodimer	NN	O	I-protein
with	NN	O	O
a	NN	O	O
novel	NN	O	O
TRE	NN	O	B-DNA
(	NN	O	O
DR+2	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

The	NN	O	O
critical	NN	O	O
region	NN	O	O
of	NN	O	O
THR	NN	O	B-protein
beta	NN	O	O
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
3	NN	O	O
amino	NN	O	O
acids	NN	O	O
in	NN	O	O
the	NN	O	O
distal	NN	O	B-protein
box	NN	O	I-protein
of	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
PPAR	NN	O	B-protein
can	NN	O	O
positively	NN	O	O
or	NN	O	O
negatively	NN	O	O
influence	NN	O	O
TH	NN	O	O
action	NN	O	O
depending	NN	O	O
on	NN	O	O
TRE	NN	O	B-DNA
structure	NN	O	O
and	NN	O	O
THR	NN	O	B-protein
isotype	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Comparison	NN	O	O
of	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
and	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
as	NN	O	O
inducers	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Several	NN	O	O
human	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
growing	NN	O	O
in	NN	O	O
vitro	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
to	NN	O	O
more	NN	O	O
mature	NN	O	O
monocyte/macrophage-like	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
-activating	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
PMA	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
PMA	NN	O	O
,	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
THP-1	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
acquire	NN	O	O
macrophage-like	NN	O	O
characteristics	NN	O	O
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
.	NN	O	O

To	NN	O	O
analyze	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
mechanisms	NN	O	O
induced	NN	O	O
by	NN	O	O
RA	NN	O	O
,	NN	O	O
we	NN	O	O
first	NN	O	O
compared	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
PMA	NN	O	O
and	NN	O	O
RA	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
which	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Both	NN	O	O
RA	NN	O	O
and	NN	O	O
PMA	NN	O	O
effectively	NN	O	O
down-regulated	NN	O	O
c-myc	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
while	NN	O	O
c-myb	NN	O	B-DNA
expression	NN	O	O
decreased	NN	O	O
only	NN	O	O
after	NN	O	O
PMA	NN	O	O
treatment	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
beta	NN	O	B-DNA
2-integrin	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
CD11a	NN	O	B-DNA
and	NN	O	O
CD11b	NN	O	B-DNA
,	NN	O	O
was	NN	O	O
clearly	NN	O	O
increased	NN	O	O
after	NN	O	O
both	NN	O	O
of	NN	O	O
these	NN	O	O
treatments	NN	O	O
.	NN	O	O

Their	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
src-family	NN	O	B-DNA
tyrosine	NN	O	I-DNA
kinase	NN	O	I-DNA
genes	NN	O	I-DNA
were	NN	O	O
different	NN	O	O
:	NN	O	O
hck	NN	O	B-DNA
expression	NN	O	O
was	NN	O	O
similarly	NN	O	O
induced	NN	O	O
by	NN	O	O
these	NN	O	O
agents	NN	O	O
but	NN	O	O
lyn	NN	O	O
expression	NN	O	O
was	NN	O	O
stronger	NN	O	O
and	NN	O	O
more	NN	O	O
rapid	NN	O	O
after	NN	O	O
RA	NN	O	O
treatment	NN	O	O
.	NN	O	O

RA	NN	O	O
also	NN	O	O
enhanced	NN	O	O
lyn	NN	O	B-RNA
mRNA	NN	O	I-RNA
production	NN	O	O
rapidly	NN	O	O
in	NN	O	O
HL-60	NN	O	B-cell_line
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
lyn	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
is	NN	O	O
common	NN	O	O
in	NN	O	O
monocytic	NN	O	O
and	NN	O	O
granulocytic	NN	O	O
maturation	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
examine	NN	O	O
whether	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
RA-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
,	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyl	NN	O	I-DNA
transferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
-reporter	NN	O	I-DNA
gene	NN	O	I-DNA
containing	NN	O	O
5	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
PMA	NN	O	O
,	NN	O	O
RA	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
any	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
thus	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
RA-induced	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
those	NN	O	O
genes	NN	O	O
described	NN	O	O
above	NN	O	O
were	NN	O	O
not	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
active	NN	O	O
v-abl	NN	O	B-protein
protein	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
blocks	NN	O	O
immunoglobulin	NN	O	B-DNA
light-chain	NN	O	I-DNA
gene	NN	O	I-DNA
rearrangement	NN	O	O
.	NN	O	O

Lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
transformed	NN	O	O
by	NN	O	O
Abelson	NN	O	O
murine	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
have	NN	O	O
provided	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
classic	NN	O	O
models	NN	O	O
for	NN	O	O
study	NN	O	O
of	NN	O	O
early	NN	O	O
B-cell	NN	O	O
development	NN	O	O
and	NN	O	O
immunoglobulin	NN	O	B-protein
rearrangement	NN	O	O
.	NN	O	O

Most	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
have	NN	O	O
rearranged	NN	O	O
their	NN	O	O
heavy-chain	NN	O	B-DNA
locus	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
their	NN	O	O
light	NN	O	B-DNA
chain	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
an	NN	O	O
active	NN	O	O
v-abl	NN	O	B-protein
protein	NN	O	I-protein
interferes	NN	O	O
with	NN	O	O
this	NN	O	O
differentiation	NN	O	O
step	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
light-chain	NN	O	B-DNA
gene	NN	O	I-DNA
structure	NN	O	I-DNA
was	NN	O	O
examined	NN	O	O
in	NN	O	O
pre-B	NN	O	B-cell_line
cells	NN	O	I-cell_line
transformed	NN	O	O
by	NN	O	O
temperature-sensitive	NN	O	O
mutants	NN	O	O
of	NN	O	O
the	NN	O	O
Abelson	NN	O	O
virus	NN	O	O
and	NN	O	O
in	NN	O	O
derivatives	NN	O	O
that	NN	O	O
survive	NN	O	O
at	NN	O	O
the	NN	O	O
nonpermissive	NN	O	O
temperature	NN	O	O
because	NN	O	O
they	NN	O	O
express	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
BCL-2	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
reveal	NN	O	O
that	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
v-abl	NN	O	B-protein
protein	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
triggers	NN	O	O
high-frequency	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
kappa	NN	O	B-DNA
and	NN	O	I-DNA
lambda	NN	O	I-DNA
light-chain	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
events	NN	O	O
are	NN	O	O
accompanied	NN	O	O
by	NN	O	O
marked	NN	O	O
increases	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
RAG-1	NN	O	B-RNA
and	NN	O	I-RNA
RAG-2	NN	O	I-RNA
RNAs	NN	O	I-RNA
.	NN	O	O

These	NN	O	O
increases	NN	O	O
occur	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
but	NN	O	O
are	NN	O	O
dependent	NN	O	O
on	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
v-abl	NN	O	B-protein
protein	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

As	NN	O	O
documented	NN	O	O
in	NN	O	O
the	NN	O	O
accompanying	NN	O	O
paper	NN	O	O
(	NN	O	O
Klug	NN	O	O
et	NN	O	O
al.	NN	O	O
,	NN	O	O
this	NN	O	O
issue	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
active	NN	O	O
v-abl	NN	O	B-protein
protein	NN	O	I-protein
also	NN	O	O
suppresses	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/rel	NN	O	I-protein
and	NN	O	O
expression	NN	O	O
controlled	NN	O	O
by	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
intron	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Together	NN	O	O
these	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
v-abl	NN	O	B-protein
protein	NN	O	I-protein
specifically	NN	O	O
interferes	NN	O	O
with	NN	O	O
light-chain	NN	O	B-DNA
gene	NN	O	I-DNA
rearrangement	NN	O	O
by	NN	O	O
suppressing	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
pathways	NN	O	O
essential	NN	O	O
for	NN	O	O
this	NN	O	O
stage	NN	O	O
of	NN	O	O
B-cell	NN	O	B-cell_type
differentiation	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
is	NN	O	O
important	NN	O	O
in	NN	O	O
regulating	NN	O	O
RAG	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Calcium	NN	O	O
signalling	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
a	NN	O	O
cyclophilin	NN	O	B-protein
B-binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
immunosuppressant	NN	O	O
drug	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
blocks	NN	O	O
a	NN	O	O
calcium-dependent	NN	O	O
signal	NN	O	O
from	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
that	NN	O	O
normally	NN	O	O
leads	NN	O	O
to	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

When	NN	O	O
bound	NN	O	O
to	NN	O	O
cyclophilin	NN	O	B-protein
,	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
binds	NN	O	O
and	NN	O	O
inactivates	NN	O	O
the	NN	O	O
key	NN	O	O
signalling	NN	O	O
intermediate	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

To	NN	O	O
identify	NN	O	O
potential	NN	O	O
cellular	NN	O	O
homologues	NN	O	O
of	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
that	NN	O	O
might	NN	O	O
regulate	NN	O	O
calcium	NN	O	O
signalling	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
cloned	NN	O	O
human	NN	O	O
genes	NN	O	O
encoding	NN	O	O
cyclophilin	NN	O	B-protein
B-binding-proteins	NN	O	I-protein
using	NN	O	O
the	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
system	NN	O	O
.	NN	O	O

One	NN	O	O
gene	NN	O	O
product	NN	O	O
,	NN	O	O
when	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
specifically	NN	O	O
induced	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
by	NN	O	O
activating	NN	O	O
the	NN	O	O
T-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
NF-AT	NN	O	B-protein
and	NN	O	O
NF-IL2A	NN	O	B-protein
.	NN	O	O

This	NN	O	O
protein	NN	O	O
,	NN	O	O
termed	NN	O	O
calcium-signal	NN	O	B-protein
modulating	NN	O	I-protein
cyclophilin	NN	O	I-protein
ligand	NN	O	I-protein
(	NN	O	O
CAML	NN	O	B-protein
)	NN	O	O
,	NN	O	O
acts	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
upstream	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
by	NN	O	O
causing	NN	O	O
an	NN	O	O
influx	NN	O	O
of	NN	O	O
calcium	NN	O	O
.	NN	O	O

CAML	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
new	NN	O	O
participant	NN	O	O
in	NN	O	O
the	NN	O	O
calcium-signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
,	NN	O	O
implicating	NN	O	O
cyclophilin	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
calcium	NN	O	O
signalling	NN	O	O
,	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
cyclosporin	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
and	NN	O	O
genomic	NN	O	O
configuration	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
,	NN	O	O
IL-3	NN	O	B-protein
,	NN	O	O
M-CSF	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
C-FMS	NN	O	B-protein
)	NN	O	O
,	NN	O	O
early	NN	O	B-DNA
growth	NN	O	I-DNA
response	NN	O	I-DNA
gene-1	NN	O	I-DNA
(	NN	O	O
EGR-1	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
M-CSF	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
primary	NN	O	O
myelodysplastic	NN	O	O
syndromes	NN	O	O
.	NN	O	O

Peripheral	NN	O	O
blood	NN	O	O
mononuclear	NN	O	O
cells	NN	O	O
from	NN	O	O
seventeen	NN	O	O
patients	NN	O	O
with	NN	O	O
primary	NN	O	O
myelodysplastic	NN	O	O
syndromes	NN	O	O
(	NN	O	O
MDS	NN	O	O
)	NN	O	O
in	NN	O	O
advanced	NN	O	O
stage	NN	O	O
were	NN	O	O
enriched	NN	O	O
for	NN	O	O
blasts	NN	O	B-cell_type
and	NN	O	O
tested	NN	O	O
for	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
karyotype	NN	O	O
,	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
genomic	NN	O	O
configuration	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-3	NN	O	B-DNA
,	NN	O	I-DNA
GM-CSF	NN	O	I-DNA
,	NN	O	I-DNA
FMS	NN	O	I-DNA
and	NN	O	I-DNA
EGR-1	NN	O	I-DNA
genes	NN	O	I-DNA
which	NN	O	O
are	NN	O	O
all	NN	O	O
located	NN	O	O
on	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
arm	NN	O	I-DNA
of	NN	O	I-DNA
chromosome	NN	O	I-DNA
5	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
that	NN	O	O
has	NN	O	O
been	NN	O	O
recently	NN	O	O
reassigned	NN	O	O
to	NN	O	O
the	NN	O	O
short	NN	O	B-DNA
arm	NN	O	I-DNA
of	NN	O	I-DNA
chromosome	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	O
lp	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
was	NN	O	O
also	NN	O	O
investigated	NN	O	O
.	NN	O	O

Aims	NN	O	O
of	NN	O	O
the	NN	O	O
study	NN	O	O
were	NN	O	O
to	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
assess	NN	O	O
the	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
in	NN	O	O
the	NN	O	O
maintenance	NN	O	O
and	NN	O	O
expansion	NN	O	O
of	NN	O	O
the	NN	O	O
neoplastic	NN	O	B-cell_line
clones	NN	O	I-cell_line
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
search	NN	O	O
for	NN	O	O
constitutional	NN	O	O
losses	NN	O	O
or	NN	O	O
rearrangements	NN	O	O
of	NN	O	O
one	NN	O	O
allele	NN	O	B-DNA
followed	NN	O	O
by	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
second	NN	O	O
allele	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
same	NN	O	O
genes	NN	O	O
in	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
latter	NN	O	O
issue	NN	O	O
was	NN	O	O
investigated	NN	O	O
by	NN	O	O
comparing	NN	O	O
,	NN	O	O
in	NN	O	O
8	NN	O	O
cases	NN	O	O
,	NN	O	O
constitutive	NN	O	B-DNA
DNA	NN	O	I-DNA
from	NN	O	O
skin	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
with	NN	O	O
leukemic	NN	O	B-DNA
DNA	NN	O	I-DNA
.	NN	O	O

Eleven	NN	O	O
of	NN	O	O
the	NN	O	O
17	NN	O	O
patients	NN	O	O
had	NN	O	O
abnormal	NN	O	O
karyotypes	NN	O	O
.	NN	O	O

The	NN	O	O
M-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
6	NN	O	O
cases	NN	O	O
and	NN	O	O
the	NN	O	O
FMS	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
EGR-1	NN	O	B-DNA
genes	NN	O	I-DNA
were	NN	O	O
expressed	NN	O	O
in	NN	O	O
2	NN	O	O
of	NN	O	O
the	NN	O	O
latter	NN	O	O
cases	NN	O	O
.	NN	O	O

An	NN	O	O
autocrine	NN	O	O
mechanism	NN	O	O
of	NN	O	O
growth	NN	O	O
could	NN	O	O
be	NN	O	O
hypothesized	NN	O	O
only	NN	O	O
for	NN	O	O
the	NN	O	O
2	NN	O	O
patients	NN	O	O
whose	NN	O	O
cells	NN	O	O
expressed	NN	O	O
both	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-DNA
and	NN	O	I-DNA
FMS	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

No	NN	O	O
germline	NN	O	O
changes	NN	O	O
or	NN	O	O
rearrangements	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
studied	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
deregulation	NN	O	O
of	NN	O	O
genes	NN	O	O
encoding	NN	O	O
for	NN	O	O
certain	NN	O	O
hemopoietic	NN	O	B-protein
growth	NN	O	I-protein
factors	NN	O	I-protein
or	NN	O	O
receptors	NN	O	O
does	NN	O	O
not	NN	O	O
seem	NN	O	O
to	NN	O	O
represent	NN	O	O
a	NN	O	O
major	NN	O	O
mechanism	NN	O	O
of	NN	O	O
MDS	NN	O	O
progression	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
human	NN	O	B-DNA
homeobox	NN	O	I-DNA
gene	NN	O	I-DNA
distantly	NN	O	O
related	NN	O	O
to	NN	O	O
proboscipedia	NN	O	B-DNA
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
and	NN	O	O
pancreatic	NN	O	O
tissues	NN	O	O
.	NN	O	O

A	NN	O	O
novel	NN	O	O
human	NN	O	B-DNA
homeobox	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
HB9	NN	O	B-DNA
,	NN	O	O
was	NN	O	O
isolated	NN	O	O
from	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
library	NN	O	I-DNA
prepared	NN	O	O
from	NN	O	O
in	NN	O	O
vitro	NN	O	B-cell_type
stimulated	NN	O	I-cell_type
human	NN	O	I-cell_type
tonsil	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
from	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
genomic	NN	O	I-DNA
library	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
HB9	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
composed	NN	O	O
of	NN	O	O
3	NN	O	O
exons	NN	O	B-DNA
spread	NN	O	O
over	NN	O	O
6	NN	O	O
kilobases	NN	O	O
of	NN	O	O
DNA	NN	O	O
.	NN	O	O

An	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
of	NN	O	O
1206	NN	O	O
nucleotides	NN	O	O
is	NN	O	O
in	NN	O	O
frame	NN	O	O
with	NN	O	O
a	NN	O	O
diverged	NN	O	B-DNA
homeodomain	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
predicted	NN	O	O
HB9	NN	O	B-protein
protein	NN	O	I-protein
has	NN	O	O
a	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
41	NN	O	O
kilodaltons	NN	O	O
and	NN	O	O
is	NN	O	O
enriched	NN	O	O
for	NN	O	O
alanine	NN	O	O
,	NN	O	O
glycine	NN	O	O
,	NN	O	O
and	NN	O	O
leucine	NN	O	O
.	NN	O	O

The	NN	O	O
HB9	NN	O	B-DNA
homeodomain	NN	O	I-DNA
is	NN	O	O
most	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-DNA
melanogaster	NN	O	I-DNA
homeobox	NN	O	I-DNA
gene	NN	O	I-DNA
proboscipedia	NN	O	B-DNA
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
poly	NN	O	B-RNA
(	NN	O	I-RNA
A	NN	O	I-RNA
)	NN	O	I-RNA
RNA	NN	O	I-RNA
purified	NN	O	O
from	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
RPMI	NN	O	I-cell_line
8226	NN	O	I-cell_line
and	NN	O	O
from	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
revealed	NN	O	O
a	NN	O	O
major	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcript	NN	O	I-RNA
of	NN	O	O
2.2	NN	O	O
kilobases	NN	O	O
.	NN	O	O

Similar	NN	O	O
analysis	NN	O	O
of	NN	O	O
poly	NN	O	B-RNA
(	NN	O	I-RNA
A	NN	O	I-RNA
)	NN	O	I-RNA
RNA	NN	O	I-RNA
from	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
adult	NN	O	O
tissues	NN	O	O
demonstrated	NN	O	O
HB9	NN	O	B-DNA
transcripts	NN	O	O
in	NN	O	O
pancreas	NN	O	O
,	NN	O	O
small	NN	O	O
intestine	NN	O	O
,	NN	O	O
and	NN	O	O
colon	NN	O	O
.	NN	O	O

Reverse	NN	O	O
transcriptase-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
examine	NN	O	O
HB9	NN	O	B-RNA
RNA	NN	O	I-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
hematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

HB9	NN	O	B-RNA
RNA	NN	O	I-RNA
transcripts	NN	O	I-RNA
were	NN	O	O
most	NN	O	O
prevalent	NN	O	O
in	NN	O	O
several	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
transcripts	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
RNA	NN	O	O
prepared	NN	O	O
from	NN	O	O
tonsil	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
studies	NN	O	O
localized	NN	O	O
them	NN	O	O
in	NN	O	O
the	NN	O	O
germinal	NN	O	O
center	NN	O	O
region	NN	O	O
of	NN	O	O
adult	NN	O	O
tonsil	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
HB9	NN	O	B-DNA
in	NN	O	O
regulating	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
and	NN	O	O
pancreatic	NN	O	O
tissues	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-protein
immunodeficiency	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
Tat	NN	O	I-protein
upregulates	NN	O	O
interleukin-2	NN	O	B-protein
secretion	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Dysregulation	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
secreted	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
AIDS	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
human	NN	O	B-protein
immunodeficiency	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
HIV-1	NN	O	I-protein
)	NN	O	I-protein
Tat	NN	O	I-protein
on	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
IL-2	NN	O	B-DNA
promoter-chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
constructs	NN	O	I-DNA
and	NN	O	O
IL-2-secreting	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
a	NN	O	O
model	NN	O	O
system	NN	O	O
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-protein
Tat	NN	O	I-protein
induced	NN	O	O
a	NN	O	O
five-	NN	O	O
to	NN	O	O
eightfold	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
stimulated	NN	O	O
with	NN	O	O
phytohemagglutinin	NN	O	B-protein
and	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
.	NN	O	O

IL-2	NN	O	B-protein
secretion	NN	O	O
was	NN	O	O
increased	NN	O	O
more	NN	O	O
than	NN	O	O
twofold	NN	O	O
in	NN	O	O
both	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
primary	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
stimulated	NN	O	O
by	NN	O	O
extracellular	NN	O	B-protein
HIV-1	NN	O	I-protein
Tat	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
suggested	NN	O	O
that	NN	O	O
Tat	NN	O	O
exerts	NN	O	O
its	NN	O	O
effect	NN	O	O
on	NN	O	O
IL-2	NN	O	B-protein
primarily	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

The	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
at	NN	O	O
positions	NN	O	O
-206	NN	O	B-DNA
to	NN	O	I-DNA
-195	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
required	NN	O	O
but	NN	O	O
not	NN	O	O
sufficient	NN	O	O
for	NN	O	O
the	NN	O	O
Tat	NN	O	B-protein
effect	NN	O	O
.	NN	O	O

The	NN	O	O
Tat	NN	O	B-protein
-mediated	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
could	NN	O	O
selectively	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
antisense	NN	O	O
tat	NN	O	O
or-unlike	NN	O	O
the	NN	O	O
analogous	NN	O	O
effect	NN	O	O
of	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
lymphotropic	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
Tax	NN	O	I-protein
-by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

The	NN	O	O
observed	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
levels	NN	O	O
might	NN	O	O
facilitate	NN	O	O
virus	NN	O	O
spread	NN	O	O
from	NN	O	O
or	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
it	NN	O	O
might	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
hypergammaglobulinemia	NN	O	O
or	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
found	NN	O	O
to	NN	O	O
be	NN	O	O
dysregulated	NN	O	O
,	NN	O	O
the	NN	O	O
T-helper	NN	O	O
cell	NN	O	O
dysfunctions	NN	O	O
observed	NN	O	O
in	NN	O	O
AIDS	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
lymphoblastoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
low-dose	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
pleiotropic	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
which	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
several	NN	O	O
specific	NN	O	O
genes	NN	O	O
.	NN	O	O

Recent	NN	O	O
reports	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
in	NN	O	O
the	NN	O	O
dose	NN	O	O
range	NN	O	O
of	NN	O	O
2-50	NN	O	O
Gy	NN	O	O
results	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
KG-1	NN	O	I-cell_line
myeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
human	NN	O	B-cell_type
B-lymphocyte	NN	O	I-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
the	NN	O	O
precise	NN	O	O
mechanism	NN	O	O
involved	NN	O	O
and	NN	O	O
the	NN	O	O
significance	NN	O	O
are	NN	O	O
not	NN	O	O
yet	NN	O	O
known	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
report	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
even	NN	O	O
lower	NN	O	O
doses	NN	O	O
of	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
,	NN	O	O
0.25-2.0	NN	O	O
Gy	NN	O	O
,	NN	O	O
are	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
EBV-transformed	NN	O	B-cell_line
244B	NN	O	I-cell_line
human	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
are	NN	O	O
in	NN	O	O
a	NN	O	O
dose	NN	O	O
range	NN	O	O
where	NN	O	O
the	NN	O	O
viability	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
remains	NN	O	O
very	NN	O	O
high	NN	O	O
.	NN	O	O

After	NN	O	O
exposure	NN	O	O
to	NN	O	O
137Cs	NN	O	O
gamma	NN	O	O
rays	NN	O	O
at	NN	O	O
a	NN	O	O
dose	NN	O	O
rate	NN	O	O
of	NN	O	O
1.17	NN	O	O
Gy/min	NN	O	O
,	NN	O	O
a	NN	O	O
maximum	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
seen	NN	O	O
at	NN	O	O
8	NN	O	O
h	NN	O	O
after	NN	O	O
a	NN	O	O
0.5-Gy	NN	O	O
exposure	NN	O	O
.	NN	O	O

Time-course	NN	O	O
studies	NN	O	O
revealed	NN	O	O
a	NN	O	O
biphasic	NN	O	O
time-dependent	NN	O	O
expression	NN	O	O
after	NN	O	O
0.5-	NN	O	O
,	NN	O	O
1-	NN	O	O
and	NN	O	O
2-Gy	NN	O	O
exposures	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
for	NN	O	O
each	NN	O	O
time	NN	O	O
examined	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
maximum	NN	O	O
after	NN	O	O
the	NN	O	O
0.5-Gy	NN	O	O
exposure	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
and	NN	O	I-protein
p65	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
subunits	NN	O	I-protein
was	NN	O	O
also	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
differentially	NN	O	O
after	NN	O	O
exposures	NN	O	O
to	NN	O	O
1.0	NN	O	O
and	NN	O	O
2.0	NN	O	O
Gy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Alternative	NN	O	O
splicing	NN	O	O
of	NN	O	O
RNA	NN	O	B-RNA
transcripts	NN	O	I-RNA
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
p105	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
gene	NN	O	I-DNA
generates	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma	NN	O	I-protein
isoforms	NN	O	I-protein
with	NN	O	O
different	NN	O	O
inhibitory	NN	O	O
activities	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
105-kDa	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
p105	NN	O	I-protein
)	NN	O	I-protein
precursor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
subunit	NN	O	O
of	NN	O	O
transcription	NN	O	O
factor	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
also	NN	O	O
encodes	NN	O	O
a	NN	O	O
p70	NN	O	B-protein
I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
607	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
of	NN	O	O
p105	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
alternative	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
generates	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma	NN	O	I-protein
isoforms	NN	O	I-protein
with	NN	O	O
properties	NN	O	O
different	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
p70	NN	O	B-protein
.	NN	O	O

One	NN	O	O
63-kDa	NN	O	B-protein
isoform	NN	O	I-protein
,	NN	O	O
termed	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma-1	NN	O	I-protein
,	NN	O	O
which	NN	O	O
lacks	NN	O	O
59	NN	O	O
amino	NN	O	O
acids	NN	O	O
C-terminal	NN	O	O
to	NN	O	O
ankyrin	NN	O	B-protein
repeat	NN	O	I-protein
7	NN	O	I-protein
,	NN	O	O
has	NN	O	O
a	NN	O	O
novel	NN	O	O
35-amino	NN	O	B-protein
acid	NN	O	I-protein
C	NN	O	I-protein
terminus	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
an	NN	O	O
alternative	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
p105	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
55-kDa	NN	O	B-protein
isoform	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma-2	NN	O	I-protein
,	NN	O	O
lacks	NN	O	O
the	NN	O	O
190	NN	O	B-protein
C-terminal	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
of	NN	O	O
p70	NN	O	B-protein
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
p70	NN	O	B-protein
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma-1	NN	O	I-protein
is	NN	O	O
found	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
and	NN	O	O
nucleus	NN	O	O
,	NN	O	O
whereas	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma-2	NN	O	I-protein
is	NN	O	O
predominantly	NN	O	O
nuclear	NN	O	O
.	NN	O	O

The	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma	NN	O	I-protein
isoforms	NN	O	I-protein
also	NN	O	O
display	NN	O	O
differences	NN	O	O
in	NN	O	O
specificity	NN	O	O
and	NN	O	O
affinity	NN	O	O
for	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

While	NN	O	O
p70	NN	O	B-protein
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma	NN	O	I-protein
inhibits	NN	O	O
p50-	NN	O	O
,	NN	O	O
p65-	NN	O	O
,	NN	O	O
and	NN	O	O
c-Rel-mediated	NN	O	O
transactivation	NN	O	O
and/or	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
both	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma-1	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma-2	NN	O	I-protein
are	NN	O	O
specific	NN	O	O
for	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
have	NN	O	O
different	NN	O	O
affinities	NN	O	O
for	NN	O	O
this	NN	O	O
subunit	NN	O	O
.	NN	O	O

The	NN	O	O
absence	NN	O	O
in	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma-1	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma-2	NN	O	I-protein
of	NN	O	O
a	NN	O	O
protein	NN	O	B-DNA
kinase	NN	O	I-DNA
A	NN	O	I-DNA
site	NN	O	I-DNA
whose	NN	O	O
phosphorylation	NN	O	O
modulates	NN	O	O
p70	NN	O	B-protein
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma	NN	O	I-protein
inhibitory	NN	O	O
activity	NN	O	O
suggests	NN	O	O
that	NN	O	O
alternative	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
may	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
generate	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma	NN	O	I-protein
isoforms	NN	O	I-protein
that	NN	O	O
respond	NN	O	O
differently	NN	O	O
to	NN	O	O
intracellular	NN	O	O
signals	NN	O	O
.	NN	O	O

-DOCSTART-	O

Structure	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GATA3	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

GATA3	NN	O	B-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
GATA	NN	O	B-protein
family	NN	O	I-protein
that	NN	O	O
is	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
T-lymphocyte	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
participate	NN	O	O
in	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
gene	NN	O	O
activation	NN	O	O
through	NN	O	O
binding	NN	O	O
to	NN	O	O
enhancers	NN	O	B-DNA
.	NN	O	O

To	NN	O	O
understand	NN	O	O
GATA3	NN	O	B-DNA
gene	NN	O	I-DNA
regulation	NN	O	O
,	NN	O	O
we	NN	O	O
cloned	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
end	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
GATA3	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GATA3	NN	O	I-DNA
gene	NN	O	I-DNA
contains	NN	O	O
six	NN	O	O
exons	NN	O	B-DNA
distributed	NN	O	O
over	NN	O	O
17	NN	O	O
kb	NN	O	O
of	NN	O	O
DNA	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
human	NN	O	B-protein
GATA3	NN	O	I-protein
zinc	NN	O	I-protein
fingers	NN	O	I-protein
are	NN	O	O
encoded	NN	O	O
by	NN	O	O
two	NN	O	O
separate	NN	O	O
exons	NN	O	B-DNA
highly	NN	O	O
conserved	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
GATA1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
no	NN	O	O
other	NN	O	O
structural	NN	O	O
homologies	NN	O	O
between	NN	O	O
these	NN	O	O
two	NN	O	O
genes	NN	O	O
can	NN	O	O
be	NN	O	O
found	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
mouse	NN	O	I-DNA
GATA3	NN	O	I-DNA
transcription	NN	O	I-DNA
units	NN	O	I-DNA
start	NN	O	O
at	NN	O	O
a	NN	O	O
major	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
promoter	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
genes	NN	O	O
revealed	NN	O	O
that	NN	O	O
they	NN	O	O
are	NN	O	O
embedded	NN	O	O
within	NN	O	O
a	NN	O	O
CpG	NN	O	B-DNA
island	NN	O	I-DNA
and	NN	O	O
share	NN	O	O
structural	NN	O	O
features	NN	O	O
often	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
housekeeping	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
DNA	NN	O	B-DNA
fragment	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GATA3	NN	O	I-DNA
transcription	NN	O	I-DNA
unit	NN	O	I-DNA
,	NN	O	O
3	NN	O	B-DNA
kb	NN	O	I-DNA
upstream	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
initiation	NN	O	B-DNA
site	NN	O	I-DNA
and	NN	O	O
4	NN	O	B-DNA
kb	NN	O	I-DNA
downstream	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
polyadenylation	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
displays	NN	O	O
T-cell	NN	O	O
specificity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
LBR	NN	O	B-protein
,	NN	O	O
an	NN	O	O
integral	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
envelope	NN	O	O
inner	NN	O	O
membrane	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
characterized	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
LBR	NN	O	B-protein
,	NN	O	O
an	NN	O	O
integral	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
envelope	NN	O	O
inner	NN	O	O
membrane	NN	O	O
.	NN	O	O

Restriction	NN	O	O
mapping	NN	O	O
shows	NN	O	O
that	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
unit	NN	O	I-DNA
spans	NN	O	O
approximately	NN	O	O
35	NN	O	O
kilobases	NN	O	O
.	NN	O	O

A	NN	O	O
transcription	NN	O	O
start	NN	O	O
site	NN	O	O
is	NN	O	O
located	NN	O	O
approximately	NN	O	O
4	NN	O	O
kilobases	NN	O	O
5	NN	O	O
'	NN	O	O
to	NN	O	O
the	NN	O	O
translation	NN	O	B-DNA
initiation	NN	O	I-DNA
codon	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
an	NN	O	O
RNA	NN	O	O
splice	NN	O	O
of	NN	O	O
3863	NN	O	O
bases	NN	O	O
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
5'-untranslated	NN	O	B-DNA
region	NN	O	I-DNA
to	NN	O	O
generate	NN	O	O
mature	NN	O	B-RNA
HeLa	NN	O	I-RNA
cell	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

5	NN	O	O
'	NN	O	O
to	NN	O	O
the	NN	O	O
identified	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
are	NN	O	O
two	NN	O	O
CCAAT	NN	O	B-DNA
sequences	NN	O	I-DNA
and	NN	O	O
potential	NN	O	B-DNA
recognition	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
including	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
AP-2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-kB	NN	O	B-protein
.	NN	O	O

There	NN	O	O
are	NN	O	O
13	NN	O	O
protein	NN	O	O
coding	NN	O	O
exons	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
LBR	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

LBR	NN	O	B-protein
's	NN	O	O
nucleoplasmic	NN	O	B-protein
domain	NN	O	I-protein
is	NN	O	O
encoded	NN	O	O
by	NN	O	O
exons	NN	O	B-DNA
1-4	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
its	NN	O	O
hydrophobic	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
with	NN	O	O
eight	NN	O	O
putative	NN	O	O
transmembrane	NN	O	B-protein
segments	NN	O	I-protein
,	NN	O	O
is	NN	O	O
encoded	NN	O	O
by	NN	O	O
exons	NN	O	B-DNA
5-13	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
hydrophobic	NN	O	B-protein
domain	NN	O	I-protein
is	NN	O	O
homologous	NN	O	O
to	NN	O	O
three	NN	O	O
yeast	NN	O	O
polypeptides	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
higher	NN	O	B-DNA
eukaryotic	NN	O	I-DNA
gene	NN	O	I-DNA
could	NN	O	O
have	NN	O	O
evolved	NN	O	O
from	NN	O	O
recombination	NN	O	O
between	NN	O	O
a	NN	O	O
gene	NN	O	O
that	NN	O	O
encoded	NN	O	O
a	NN	O	O
soluble	NN	O	B-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
a	NN	O	O
membrane	NN	O	B-DNA
protein	NN	O	I-DNA
gene	NN	O	I-DNA
similar	NN	O	O
to	NN	O	O
those	NN	O	O
in	NN	O	O
yeast	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
are	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
structural	NN	O	O
organization	NN	O	O
of	NN	O	O
a	NN	O	O
vertebrate	NN	O	B-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
an	NN	O	O
integral	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
envelope	NN	O	O
that	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
polypeptides	NN	O	O
conserved	NN	O	O
in	NN	O	O
evolution	NN	O	O
.	NN	O	O

-DOCSTART-	O

Retinoic	NN	O	O
acid-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
CD38	NN	O	B-protein
antigen	NN	O	I-protein
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor-alpha	NN	O	I-protein
.	NN	O	O

CD38	NN	O	B-protein
is	NN	O	O
a	NN	O	O
leukocyte	NN	O	B-protein
differentiation	NN	O	I-protein
antigen	NN	O	I-protein
that	NN	O	O
has	NN	O	O
been	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
phenotypic	NN	O	O
marker	NN	O	O
of	NN	O	O
different	NN	O	O
subpopulations	NN	O	O
of	NN	O	O
T-	NN	O	B-cell_type
and	NN	O	I-cell_type
B-	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
CD38	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
during	NN	O	O
early	NN	O	O
stages	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Virtually	NN	O	O
no	NN	O	O
information	NN	O	O
is	NN	O	O
available	NN	O	O
on	NN	O	O
regulation	NN	O	O
and	NN	O	O
functions	NN	O	O
of	NN	O	O
CD38	NN	O	B-protein
.	NN	O	O

Recently	NN	O	O
we	NN	O	O
reported	NN	O	O
that	NN	O	O
all-trans-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
and	NN	O	O
highly	NN	O	O
specific	NN	O	O
inducer	NN	O	O
of	NN	O	O
CD38	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
promyelocytic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
ATRA-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
CD38	NN	O	B-protein
antigen	NN	O	I-protein
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
retinoic	NN	O	B-protein
acid-alpha	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
.	NN	O	O

ATRA	NN	O	O
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
CD38	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
a	NN	O	O
mutant	NN	O	B-cell_line
subclone	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
HL-60	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
designated	NN	O	O
HL-60R	NN	O	B-cell_line
)	NN	O	O
that	NN	O	O
is	NN	O	O
relatively	NN	O	O
resistant	NN	O	O
to	NN	O	O
ATRA-induced	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Retroviral	NN	O	O
vector-mediated	NN	O	O
transduction	NN	O	O
of	NN	O	O
RA	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
into	NN	O	O
this	NN	O	O
HL-60R	NN	O	B-cell_line
subclone	NN	O	I-cell_line
completely	NN	O	O
restored	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
to	NN	O	O
ATRA	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
express	NN	O	O
CD38	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
CD38	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
not	NN	O	O
inducible	NN	O	O
by	NN	O	O
ATRA	NN	O	O
in	NN	O	O
HL-60R	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
functional	NN	O	O
RAR	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
RAR	NN	O	B-protein
gamma	NN	O	I-protein
,	NN	O	O
or	NN	O	O
RXR	NN	O	B-protein
alpha	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
CD38	NN	O	B-protein
in	NN	O	O
acute	NN	O	B-cell_type
promyelocytic	NN	O	I-cell_type
and	NN	O	I-cell_type
acute	NN	O	I-cell_type
myeloblastic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
independent	NN	O	O
of	NN	O	O
ATRA-induced	NN	O	O
cytodifferentiation	NN	O	O
.	NN	O	O

Following	NN	O	O
culture	NN	O	O
with	NN	O	O
ATRA	NN	O	O
,	NN	O	O
increased	NN	O	O
CD38	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
were	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
CD34+	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
on	NN	O	O
normal	NN	O	B-cell_type
circulating	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
.	NN	O	O

From	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
CD38	NN	O	B-protein
is	NN	O	O
ATRA	NN	O	O
inducible	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
normal	NN	O	B-cell_type
CD34+	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
effect	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
differentiation	NN	O	O
and	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
in	NN	O	O
HL-60	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
similar	NN	O	O
role	NN	O	O
for	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
in	NN	O	O
CD38	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
other	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Some	NN	O	O
antioxidants	NN	O	O
inhibit	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
co-ordinate	NN	O	O
fashion	NN	O	O
,	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
,	NN	O	O
IL-beta	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-6	NN	O	B-protein
by	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Some	NN	O	O
antioxidants	NN	O	O
,	NN	O	O
including	NN	O	O
butylated	NN	O	O
hydroxyanisole	NN	O	O
(	NN	O	O
BHA	NN	O	O
)	NN	O	O
,	NN	O	O
tetrahydropapaveroline	NN	O	O
(	NN	O	O
THP	NN	O	O
)	NN	O	O
,	NN	O	O
nordihydroguiauretic	NN	O	O
acid	NN	O	O
,	NN	O	O
and	NN	O	O
10	NN	O	O
,	NN	O	O
11-dihydroxyaporphine	NN	O	O
(	NN	O	O
DHA	NN	O	O
)	NN	O	O
,	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
potent	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
-alpha	NN	O	I-protein
,	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
and	NN	O	O
IL-6	NN	O	B-protein
by	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
stimulated	NN	O	O
by	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
(	NN	O	O
IC50s	NN	O	O
in	NN	O	O
the	NN	O	O
low	NN	O	O
micromolar	NN	O	O
range	NN	O	O
)	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
was	NN	O	O
gene	NN	O	O
selective	NN	O	O
and	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
general	NN	O	O
effects	NN	O	O
on	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
by	NN	O	O
PBMC	NN	O	B-cell_type
was	NN	O	O
observed	NN	O	O
also	NN	O	O
when	NN	O	O
other	NN	O	O
inducers	NN	O	O
were	NN	O	O
used	NN	O	O
(	NN	O	O
staphylococci	NN	O	O
,	NN	O	O
silica	NN	O	O
,	NN	O	O
zymosan	NN	O	O
)	NN	O	O
.	NN	O	O

Much	NN	O	O
higher	NN	O	O
concentrations	NN	O	O
of	NN	O	O
other	NN	O	O
antioxidants	NN	O	O
--	NN	O	O
including	NN	O	O
ascorbic	NN	O	O
acid	NN	O	O
,	NN	O	O
trolox	NN	O	O
,	NN	O	O
alpha-tocopherol	NN	O	O
,	NN	O	O
butylated	NN	O	O
hydroxytoluene	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
5-lipoxygenase	NN	O	O
inhibitor	NN	O	O
zileuton	NN	O	O
--	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

The	NN	O	O
active	NN	O	O
compounds	NN	O	O
did	NN	O	O
not	NN	O	O
inhibit	NN	O	O
IL-1	NN	O	B-protein
-induced	NN	O	O
production	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
in	NN	O	O
fibroblasts	NN	O	B-cell_type
,	NN	O	O
showing	NN	O	O
the	NN	O	O
cell	NN	O	O
selectivity	NN	O	O
of	NN	O	O
the	NN	O	O
effect	NN	O	O
.	NN	O	O

Antioxidant-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
was	NN	O	O
correlated	NN	O	O
with	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
corresponding	NN	O	O
messenger	NN	O	B-RNA
RNAs	NN	O	I-RNA
.	NN	O	O

Nuclear	NN	O	O
run-on	NN	O	O
experiments	NN	O	O
showed	NN	O	O
that	NN	O	O
THP	NN	O	O
inhibited	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1	NN	O	B-DNA
beta	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

THP	NN	O	O
decreased	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
detected	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
PBMC	NN	O	B-cell_type
cultured	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
LPS	NN	O	O
.	NN	O	O

THP	NN	O	O
and	NN	O	O
DHA	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
in	NN	O	O
the	NN	O	O
circulation	NN	O	O
of	NN	O	O
mice	NN	O	O
following	NN	O	O
LPS	NN	O	O
injection	NN	O	O
.	NN	O	O

Thus	NN	O	O
antioxidants	NN	O	O
vary	NN	O	O
widely	NN	O	O
in	NN	O	O
potency	NN	O	O
as	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
for	NN	O	O
pro-inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

Coordinate	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
for	NN	O	O
inflammatory	NN	O	O
cytokines	NN	O	B-protein
could	NN	O	O
provide	NN	O	O
a	NN	O	O
strategy	NN	O	O
for	NN	O	O
therapy	NN	O	O
of	NN	O	O
diseases	NN	O	O
with	NN	O	O
inflammatory	NN	O	O
pathogenesis	NN	O	O
and	NN	O	O
for	NN	O	O
septic	NN	O	O
shock	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
interleukin-4-induced	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
:	NN	O	O
IL-4	NN	O	B-protein
Stat	NN	O	I-protein
.	NN	O	O

Interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
immunomodulatory	NN	O	O
cytokine	NN	O	O
secreted	NN	O	O
by	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
basophils	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
modulating	NN	O	O
the	NN	O	O
balance	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
helper	NN	O	I-cell_type
(	NN	O	I-cell_type
Th	NN	O	I-cell_type
)	NN	O	I-cell_type
cell	NN	O	I-cell_type
subsets	NN	O	I-cell_type
,	NN	O	O
favoring	NN	O	O
expansion	NN	O	O
of	NN	O	O
the	NN	O	O
Th2	NN	O	B-cell_type
lineage	NN	O	I-cell_type
relative	NN	O	O
to	NN	O	O
Th1	NN	O	B-cell_type
.	NN	O	O

Imbalance	NN	O	O
of	NN	O	O
these	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
subsets	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
immunological	NN	O	O
diseases	NN	O	O
including	NN	O	O
allergy	NN	O	O
,	NN	O	O
inflammation	NN	O	O
,	NN	O	O
and	NN	O	O
autoimmune	NN	O	O
disease	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
may	NN	O	O
mediate	NN	O	O
its	NN	O	O
biological	NN	O	O
effects	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
activating	NN	O	O
a	NN	O	O
tyrosine-phosphorylated	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

This	NN	O	O
protein	NN	O	O
has	NN	O	O
now	NN	O	O
been	NN	O	O
purified	NN	O	O
and	NN	O	O
its	NN	O	O
encoding	NN	O	O
gene	NN	O	O
cloned	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
the	NN	O	O
primary	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
indicates	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
Stat	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
hereby	NN	O	O
designated	NN	O	O
IL-4	NN	O	B-protein
Stat	NN	O	I-protein
.	NN	O	O

Study	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
activities	NN	O	O
of	NN	O	O
phosphotyrosine-containing	NN	O	O
peptides	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	I-protein
the	NN	O	I-protein
IL-4	NN	O	I-protein
receptor	NN	O	I-protein
provided	NN	O	O
evidence	NN	O	O
for	NN	O	O
direct	NN	O	O
coupling	NN	O	O
of	NN	O	O
receptor	NN	O	B-protein
and	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
during	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
Stat	NN	O	I-protein
activation	NN	O	O
cycle	NN	O	O
.	NN	O	O

Such	NN	O	O
observations	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
Stat	NN	O	I-protein
has	NN	O	O
the	NN	O	O
same	NN	O	O
functional	NN	O	B-protein
domain	NN	O	I-protein
for	NN	O	O
both	NN	O	O
receptor	NN	O	O
coupling	NN	O	O
and	NN	O	O
dimerization	NN	O	O
.	NN	O	O

-DOCSTART-	O

Evaluation	NN	O	O
of	NN	O	O
the	NN	O	O
respiratory	NN	O	O
epithelium	NN	O	O
of	NN	O	O
normals	NN	O	O
and	NN	O	O
individuals	NN	O	O
with	NN	O	O
cystic	NN	O	O
fibrosis	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
adenovirus	NN	O	B-DNA
E1a	NN	O	I-DNA
sequences	NN	O	I-DNA
relevant	NN	O	O
to	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
E1a-	NN	O	O
adenovirus	NN	O	O
vectors	NN	O	O
for	NN	O	O
gene	NN	O	O
therapy	NN	O	O
for	NN	O	O
the	NN	O	O
respiratory	NN	O	O
manifestations	NN	O	O
of	NN	O	O
cystic	NN	O	O
fibrosis	NN	O	O
.	NN	O	O

Lung	NN	O	O
disease	NN	O	O
associated	NN	O	O
with	NN	O	O
disorders	NN	O	O
such	NN	O	O
as	NN	O	O
cystic	NN	O	O
fibrosis	NN	O	O
(	NN	O	O
CF	NN	O	O
)	NN	O	O
may	NN	O	O
be	NN	O	O
amenable	NN	O	O
to	NN	O	O
somatic	NN	O	O
gene	NN	O	O
therapy	NN	O	O
in	NN	O	O
which	NN	O	O
there	NN	O	O
is	NN	O	O
delivery	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	B-DNA
gene	NN	O	I-DNA
directly	NN	O	O
to	NN	O	O
the	NN	O	O
respiratory	NN	O	O
epithelium	NN	O	O
using	NN	O	O
E1a-	NN	O	O
adenovirus	NN	O	O
(	NN	O	O
Ad	NN	O	O
)	NN	O	O
type	NN	O	O
2-	NN	O	O
or	NN	O	O
5-based	NN	O	O
vectors	NN	O	O
.	NN	O	O

For	NN	O	O
safety	NN	O	O
reasons	NN	O	O
,	NN	O	O
the	NN	O	O
Ad	NN	O	O
vectors	NN	O	O
are	NN	O	O
rendered	NN	O	O
replication	NN	O	O
deficient	NN	O	O
by	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
E1a	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

Because	NN	O	O
there	NN	O	O
is	NN	O	O
the	NN	O	O
theoretical	NN	O	O
possibility	NN	O	O
of	NN	O	O
an	NN	O	O
E1a-	NN	O	O
replication-deficient	NN	O	O
vector	NN	O	O
replicating	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
recombination	NN	O	O
or	NN	O	O
complementation	NN	O	O
with	NN	O	O
Ad	NN	O	B-DNA
2/5	NN	O	I-DNA
E1a	NN	O	I-DNA
sequences	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
the	NN	O	O
target	NN	O	O
cell	NN	O	O
,	NN	O	O
this	NN	O	O
study	NN	O	O
is	NN	O	O
directed	NN	O	O
toward	NN	O	O
evaluating	NN	O	O
respiratory	NN	O	O
epithelium	NN	O	O
of	NN	O	O
normals	NN	O	O
and	NN	O	O
individuals	NN	O	O
with	NN	O	O
CF	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
E1a	NN	O	O
sequences	NN	O	O
.	NN	O	O

Using	NN	O	O
Ad	NN	O	B-DNA
2/5	NN	O	I-DNA
E1a-specific	NN	O	I-DNA
primers	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
to	NN	O	O
evaluate	NN	O	O
DNA	NN	O	O
recovered	NN	O	O
from	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
nasal	NN	O	O
and	NN	O	O
bronchial	NN	O	O
epithelium	NN	O	O
recovered	NN	O	O
by	NN	O	O
brushing	NN	O	O
,	NN	O	O
E1a	NN	O	B-DNA
sequences	NN	O	I-DNA
were	NN	O	O
detected	NN	O	O
in	NN	O	O
respiratory	NN	O	O
epithelium	NN	O	O
of	NN	O	O
19	NN	O	O
of	NN	O	O
91	NN	O	O
normals	NN	O	O
(	NN	O	O
21	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
E1a-positive	NN	O	O
samples	NN	O	O
,	NN	O	O
the	NN	O	O
average	NN	O	O
of	NN	O	O
E1a	NN	O	O
copy	NN	O	O
number	NN	O	O
was	NN	O	O
55	NN	O	O
+/-	NN	O	O
18/10	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
recovered	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
CF	NN	O	O
individuals	NN	O	O
,	NN	O	O
7	NN	O	O
of	NN	O	O
52	NN	O	O
(	NN	O	O
13	NN	O	O
%	NN	O	O
)	NN	O	O
had	NN	O	O
detectable	NN	O	O
E1a	NN	O	B-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
respiratory	NN	O	O
epithelium	NN	O	O
,	NN	O	O
with	NN	O	O
E1a	NN	O	O
copy	NN	O	O
number	NN	O	O
in	NN	O	O
the	NN	O	O
positive	NN	O	O
samples	NN	O	O
of	NN	O	O
80	NN	O	O
+/-	NN	O	O
21/10	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
recovered	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
detectable	NN	O	O
Ad	NN	O	B-DNA
2/5	NN	O	I-DNA
E1a	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
respiratory	NN	O	O
epithelium	NN	O	O
of	NN	O	O
a	NN	O	O
small	NN	O	O
percentage	NN	O	O
of	NN	O	O
normals	NN	O	O
and	NN	O	O
individuals	NN	O	O
with	NN	O	O
CF	NN	O	O
.	NN	O	O

Because	NN	O	O
of	NN	O	O
the	NN	O	O
theoretical	NN	O	O
potential	NN	O	O
of	NN	O	O
such	NN	O	O
sequences	NN	O	O
supporting	NN	O	O
replication	NN	O	O
of	NN	O	O
E1a-	NN	O	O
Ad	NN	O	O
vectors	NN	O	O
,	NN	O	O
human	NN	O	O
gene	NN	O	O
therapy	NN	O	O
protocols	NN	O	O
for	NN	O	O
CF	NN	O	O
utilizing	NN	O	O
such	NN	O	O
vectors	NN	O	O
should	NN	O	O
consider	NN	O	O
evaluating	NN	O	O
study	NN	O	O
individuals	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Ad	NN	O	B-DNA
2/5	NN	O	I-DNA
E1a	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
respiratory	NN	O	O
epithelium	NN	O	O
.	NN	O	O

-DOCSTART-	O

Leiomyosarcoma	NN	O	O
of	NN	O	O
the	NN	O	O
vulva	NN	O	O
:	NN	O	O
report	NN	O	O
of	NN	O	O
a	NN	O	O
case	NN	O	O
.	NN	O	O

A	NN	O	O
52-year-old	NN	O	O
female	NN	O	O
presented	NN	O	O
with	NN	O	O
a	NN	O	O
progressively	NN	O	O
enlarging	NN	O	O
vulvar	NN	O	O
mass	NN	O	O
.	NN	O	O

Pathological	NN	O	O
evaluation	NN	O	O
revealed	NN	O	O
a	NN	O	O
high-grade	NN	O	O
vulvar	NN	O	O
leiomyosarcoma	NN	O	O
.	NN	O	O

Immunohistochemical	NN	O	O
and	NN	O	O
ultrastructural	NN	O	O
studies	NN	O	O
were	NN	O	O
performed	NN	O	O
to	NN	O	O
support	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
effort	NN	O	O
to	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
biology	NN	O	O
of	NN	O	O
this	NN	O	O
tumor	NN	O	O
additional	NN	O	O
immunohistochemical	NN	O	O
studies	NN	O	O
for	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
product	NN	O	I-protein
of	NN	O	O
p53	NN	O	B-DNA
tumor	NN	O	I-DNA
suppressor	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
type	NN	O	O
of	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
infiltrating	NN	O	O
the	NN	O	O
tumor	NN	O	O
were	NN	O	O
performed	NN	O	O
.	NN	O	O

Tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
showed	NN	O	O
an	NN	O	O
overexpression	NN	O	O
of	NN	O	O
p53	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
were	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
-positive	NN	O	O
.	NN	O	O

Macrophages	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
infiltrated	NN	O	O
the	NN	O	O
tumor	NN	O	O
in	NN	O	O
moderate	NN	O	O
numbers	NN	O	O
with	NN	O	O
occasional	NN	O	O
lymphoid	NN	O	O
aggregate	NN	O	O
formation	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
attempt	NN	O	O
to	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
biology	NN	O	O
of	NN	O	O
these	NN	O	O
tumors	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stimulation	NN	O	O
of	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
by	NN	O	O
leukemia	NN	O	B-protein
inhibitory	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

This	NN	O	O
study	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
leukemia	NN	O	B-protein
inhibitory	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
LIF	NN	O	B-protein
)	NN	O	O
on	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
replication	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
(	NN	O	O
MNP	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

LIF	NN	O	B-protein
induced	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
increase	NN	O	O
in	NN	O	O
p24	NN	O	B-protein
antigen	NN	O	I-protein
production	NN	O	O
in	NN	O	O
the	NN	O	O
chronically	NN	O	B-cell_line
infected	NN	O	I-cell_line
promonocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U1	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
magnitude	NN	O	O
and	NN	O	O
time	NN	O	O
kinetics	NN	O	O
of	NN	O	O
the	NN	O	O
LIF	NN	O	B-protein
effects	NN	O	O
were	NN	O	O
similar	NN	O	O
to	NN	O	O
interleukin	NN	O	B-protein
1	NN	O	I-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
other	NN	O	O
cytokines	NN	O	O
known	NN	O	O
to	NN	O	O
induce	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
this	NN	O	O
cell	NN	O	O
line	NN	O	O
.	NN	O	O

To	NN	O	O
characterize	NN	O	O
mechanisms	NN	O	O
responsible	NN	O	O
for	NN	O	O
these	NN	O	O
LIF	NN	O	B-protein
effects	NN	O	O
,	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	B-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kB	NN	O	I-protein
,	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
,	NN	O	O
and	NN	O	O
potential	NN	O	O
interactions	NN	O	O
with	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

LIF	NN	O	B-protein
increased	NN	O	O
steady-state	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV	NN	O	B-RNA
mRNA	NN	O	I-RNA
at	NN	O	O
2.0	NN	O	O
,	NN	O	O
4.3	NN	O	O
,	NN	O	O
and	NN	O	O
9.2	NN	O	O
kB	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
detectable	NN	O	O
by	NN	O	O
24	NN	O	O
h	NN	O	O
and	NN	O	O
persisted	NN	O	O
until	NN	O	O
72	NN	O	O
h	NN	O	O
.	NN	O	O

The	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
NF-kB	NN	O	I-protein
is	NN	O	O
a	NN	O	O
central	NN	O	O
mediator	NN	O	O
in	NN	O	O
cytokine	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
.	NN	O	O

NF-kB	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
higher	NN	O	O
in	NN	O	O
unstimulated	NN	O	B-cell_line
U1	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
parent	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U937	NN	O	B-cell_line
.	NN	O	O

In	NN	O	O
both	NN	O	O
cell	NN	O	O
lines	NN	O	O
LIF	NN	O	B-protein
increased	NN	O	O
NF-kB	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
NF-kB	NN	O	B-protein
and	NN	O	O
HIV	NN	O	O
replication	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
are	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
dependent	NN	O	O
on	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
.	NN	O	O

The	NN	O	O
oxygen	NN	O	O
radical	NN	O	O
scavenger	NN	O	O
N-acetyl-L-cysteine	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
nitric	NN	O	B-protein
oxide	NN	O	I-protein
synthase	NN	O	I-protein
,	NN	O	O
inhibited	NN	O	O
LIF	NN	O	B-protein
-induced	NN	O	O
HIV	NN	O	O
replication	NN	O	O
.	NN	O	O

LIF	NN	O	B-protein
induces	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	O
but	NN	O	O
its	NN	O	O
effects	NN	O	O
on	NN	O	O
HIV	NN	O	O
replication	NN	O	O
were	NN	O	O
not	NN	O	O
inhibited	NN	O	O
by	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	O
IL-1	NN	O	B-protein
,	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
or	NN	O	O
IL-6	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
identify	NN	O	O
LIF	NN	O	B-protein
as	NN	O	O
a	NN	O	O
stimulus	NN	O	O
of	NN	O	O
HIV	NN	O	O
replication	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
growth	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
B104	NN	O	B-cell_line
,	NN	O	O
by	NN	O	O
anti-MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
growth	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
B104	NN	O	B-cell_line
,	NN	O	O
by	NN	O	O
anti-MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antibodies	NN	O	I-protein
(	NN	O	O
Ab	NN	O	B-protein
)	NN	O	O
were	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
in	NN	O	O
anti-IgM	NN	O	B-protein
Ab	NN	O	I-protein
-induced	NN	O	O
B104	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
.	NN	O	O

Two	NN	O	O
anti-MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
Ab	NN	O	I-protein
,	NN	O	O
L227	NN	O	B-protein
and	NN	O	O
2.06	NN	O	B-protein
,	NN	O	O
inhibited	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
B104	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
2.06	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
L227	NN	O	B-protein
,	NN	O	O
needed	NN	O	O
to	NN	O	O
be	NN	O	O
further	NN	O	O
cross-linked	NN	O	O
with	NN	O	O
a	NN	O	O
goat	NN	O	B-protein
anti-mouse	NN	O	I-protein
IgG	NN	O	I-protein
Ab	NN	O	I-protein
(	NN	O	O
GAM	NN	O	B-protein
)	NN	O	O
to	NN	O	O
show	NN	O	O
the	NN	O	O
effect	NN	O	O
.	NN	O	O

L227	NN	O	B-protein
induced	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
intracellular	NN	O	O
free	NN	O	O
Ca2+	NN	O	O
concentration	NN	O	O
(	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
)	NN	O	O
from	NN	O	O
the	NN	O	O
intracellular	NN	O	O
pool	NN	O	O
and	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
phosphatidyl	NN	O	O
inositol	NN	O	O
turnover	NN	O	O
,	NN	O	O
or	NN	O	O
expression	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
whereas	NN	O	O
2.06	NN	O	B-protein
plus	NN	O	O
GAM	NN	O	B-protein
induced	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
from	NN	O	O
both	NN	O	O
the	NN	O	O
intracellular	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
the	NN	O	O
extracellular	NN	O	O
pools	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
of	NN	O	O
B104	NN	O	B-cell_line
cell	NN	O	I-cell_line
growth	NN	O	O
induced	NN	O	O
by	NN	O	O
anti-MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
Ab	NN	O	I-protein
was	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-independent	NN	O	O
and	NN	O	O
not	NN	O	O
inhibited	NN	O	O
by	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
or	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
arrest	NN	O	O
at	NN	O	O
the	NN	O	O
G2/M	NN	O	O
interphase	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
.	NN	O	O

These	NN	O	O
features	NN	O	O
are	NN	O	O
very	NN	O	O
different	NN	O	O
from	NN	O	O
those	NN	O	O
observed	NN	O	O
in	NN	O	O
B104	NN	O	B-cell_line
cell	NN	O	I-cell_line
death	NN	O	O
induced	NN	O	O
by	NN	O	O
anti-IgM	NN	O	B-protein
Ab	NN	O	I-protein
.	NN	O	O

Neither	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
nor	NN	O	O
the	NN	O	O
morphology	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
transduced	NN	O	O
the	NN	O	O
negative	NN	O	O
signals	NN	O	O
through	NN	O	O
intracellular	NN	O	O
mechanisms	NN	O	O
different	NN	O	O
from	NN	O	O
those	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
surface	NN	O	O
IgM	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
block	NN	O	O
for	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3-mediated	NN	O	O
gene	NN	O	O
regulation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Elements	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
steroid	NN	O	O
hormone	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
)	NN	O	O
to	NN	O	O
induce	NN	O	O
a	NN	O	O
biological	NN	O	O
response	NN	O	O
include	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
specific	NN	O	O
intracellular	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
vitamin	NN	O	B-protein
D3	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
)	NN	O	O
and	NN	O	O
modulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
via	NN	O	O
hormone-activated	NN	O	O
receptor	NN	O	O
binding	NN	O	O
to	NN	O	O
regulatory	NN	O	B-protein
regions	NN	O	I-protein
of	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
parameters	NN	O	O
were	NN	O	O
examined	NN	O	O
in	NN	O	O
normal	NN	O	O
and	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus-immortalized	NN	O	I-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
compared	NN	O	O
with	NN	O	O
1	NN	O	B-cell_line
alpha	NN	O	I-cell_line
,	NN	O	I-cell_line
25-	NN	O	I-cell_line
(	NN	O	I-cell_line
OH	NN	O	I-cell_line
)	NN	O	I-cell_line
2D3-responsive	NN	O	I-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
monocytic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
resting	NN	O	O
tonsillar	NN	O	O
B	NN	O	O
cells	NN	O	O
did	NN	O	O
not	NN	O	O
express	NN	O	O
VDR	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
activation	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
with	NN	O	O
interleukin-4	NN	O	B-protein
induced	NN	O	O
VDR	NN	O	B-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
exogenously	NN	O	O
supplemented	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
.	NN	O	O

As	NN	O	O
indicators	NN	O	O
of	NN	O	O
hormone-mediated	NN	O	O
gene	NN	O	O
regulation	NN	O	O
we	NN	O	O
analyzed	NN	O	O
modulation	NN	O	O
of	NN	O	O
CD23	NN	O	B-protein
,	NN	O	O
a	NN	O	O
common	NN	O	B-protein
B	NN	O	I-protein
cell/monocyte	NN	O	I-protein
surface	NN	O	I-protein
antigen	NN	O	I-protein
,	NN	O	O
and	NN	O	O
24-hydroxylase	NN	O	B-protein
.	NN	O	O

1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
inhibited	NN	O	O
CD23	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
yet	NN	O	O
failed	NN	O	O
to	NN	O	O
modulate	NN	O	O
CD23	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
induced	NN	O	O
24-hydroxylase	NN	O	B-protein
mRNA	NN	O	O
expression	NN	O	O
and	NN	O	O
metabolic	NN	O	O
activity	NN	O	O
in	NN	O	O
both	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
lectin-activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
yet	NN	O	O
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
24-hydroxylase	NN	O	B-RNA
mRNA	NN	O	I-RNA
or	NN	O	O
its	NN	O	O
metabolic	NN	O	O
activity	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
although	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
can	NN	O	O
express	NN	O	O
VDR	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	B-protein
,	NN	O	O
they	NN	O	O
exhibit	NN	O	O
a	NN	O	O
functional	NN	O	O
block	NN	O	O
for	NN	O	O
vitamin	NN	O	O
D-dependent	NN	O	O
gene	NN	O	O
regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Positive	NN	O	O
and	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
:	NN	O	O
role	NN	O	O
of	NN	O	O
multiple	NN	O	O
regulatory	NN	O	O
sites	NN	O	O
.	NN	O	O

Interleukin	NN	O	O
2	NN	O	O
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
lymphokine	NN	O	B-protein
required	NN	O	O
in	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
proliferation	NN	O	O
,	NN	O	O
clonal	NN	O	O
expansion	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
displays	NN	O	O
both	NN	O	O
T	NN	O	O
cell	NN	O	O
specific	NN	O	O
and	NN	O	O
inducible	NN	O	O
expression	NN	O	O
:	NN	O	O
it	NN	O	O
is	NN	O	O
only	NN	O	O
expressed	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
after	NN	O	O
antigenic	NN	O	O
or	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Several	NN	O	O
cis-acting	NN	O	B-DNA
regulatory	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
analysed	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
these	NN	O	O
cis-acting	NN	O	B-DNA
regulatory	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
native	NN	O	O
IL-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
promoter	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
(	NN	O	O
-276	NN	O	B-DNA
to	NN	O	I-DNA
-261	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
octamer	NN	O	I-DNA
(	NN	O	O
-256	NN	O	B-DNA
to	NN	O	I-DNA
-248	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
octamer	NN	O	I-DNA
(	NN	O	O
-75	NN	O	B-DNA
to	NN	O	I-DNA
-66	NN	O	I-DNA
)	NN	O	O
sites	NN	O	O
not	NN	O	O
only	NN	O	O
act	NN	O	O
as	NN	O	O
enhancers	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cellular	NN	O	O
stimulation	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
have	NN	O	O
a	NN	O	O
silencing	NN	O	O
effect	NN	O	O
on	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
resting	NN	O	O
cells	NN	O	O
.	NN	O	O

Two	NN	O	O
other	NN	O	O
sites	NN	O	O
display	NN	O	O
disparate	NN	O	O
effects	NN	O	O
on	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
different	NN	O	O
T	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
:	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
purine	NN	O	I-DNA
box	NN	O	I-DNA
(	NN	O	O
-291	NN	O	B-DNA
to	NN	O	I-DNA
-277	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
purine	NN	O	I-DNA
box	NN	O	I-DNA
sites	NN	O	I-DNA
(	NN	O	O
-145	NN	O	B-DNA
to	NN	O	I-DNA
-128	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
(	NN	O	I-DNA
-186	NN	O	I-DNA
to	NN	O	I-DNA
-176	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
(	NN	O	O
-206	NN	O	B-DNA
to	NN	O	I-DNA
-195	NN	O	I-DNA
)	NN	O	O
respond	NN	O	O
to	NN	O	O
different	NN	O	O
cellular	NN	O	O
activation	NN	O	O
in	NN	O	O
EL4	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
mediated	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
PMA	NN	O	O
stimulation	NN	O	O
while	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
responded	NN	O	O
to	NN	O	O
IL-1	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
is	NN	O	O
a	NN	O	O
complex	NN	O	O
process	NN	O	O
and	NN	O	O
multiple	NN	O	O
cis-acting	NN	O	B-DNA
regulatory	NN	O	I-DNA
sites	NN	O	I-DNA
interact	NN	O	O
to	NN	O	O
exert	NN	O	O
different	NN	O	O
effects	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
representative	NN	O	O
of	NN	O	O
alternative	NN	O	O
stages	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Sp1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
critical	NN	O	O
factor	NN	O	O
for	NN	O	O
the	NN	O	O
monocytic	NN	O	O
specific	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-protein
CD14	NN	O	I-protein
.	NN	O	O

CD14	NN	O	B-protein
is	NN	O	O
a	NN	O	O
membrane	NN	O	B-protein
glycoprotein	NN	O	I-protein
expressed	NN	O	O
specifically	NN	O	O
on	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
its	NN	O	O
expression	NN	O	O
is	NN	O	O
markedly	NN	O	O
increased	NN	O	O
during	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
monocyte	NN	O	B-cell_type
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
study	NN	O	O
CD14	NN	O	B-DNA
gene	NN	O	I-DNA
regulation	NN	O	O
,	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD14	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
cloned	NN	O	O
from	NN	O	O
a	NN	O	O
partial	NN	O	O
EcoRI	NN	O	B-DNA
digested	NN	O	I-DNA
chromosome	NN	O	I-DNA
5	NN	O	I-DNA
library	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
5.5-kilobase	NN	O	B-DNA
genomic	NN	O	I-DNA
clone	NN	O	I-DNA
contained	NN	O	O
the	NN	O	O
full-length	NN	O	O
CD14	NN	O	B-DNA
coding	NN	O	I-DNA
sequence	NN	O	I-DNA
and	NN	O	O
4.2	NN	O	B-DNA
kilobases	NN	O	I-DNA
of	NN	O	I-DNA
5'-upstream	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

One	NN	O	O
major	NN	O	B-DNA
and	NN	O	I-DNA
one	NN	O	I-DNA
minor	NN	O	I-DNA
transcription	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
101	NN	O	B-DNA
and	NN	O	I-DNA
130	NN	O	I-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
(	NN	O	I-DNA
bp	NN	O	I-DNA
)	NN	O	I-DNA
upstream	NN	O	I-DNA
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
from	NN	O	O
the	NN	O	O
protein	NN	O	B-DNA
translation	NN	O	I-DNA
start	NN	O	I-DNA
ATG	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
DNA	NN	O	B-DNA
fragment	NN	O	I-DNA
containing	NN	O	O
128	NN	O	B-DNA
bp	NN	O	I-DNA
of	NN	O	I-DNA
upstream	NN	O	I-DNA
sequence	NN	O	I-DNA
had	NN	O	O
strong	NN	O	O
,	NN	O	O
monocyte-specific	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
CD14	NN	O	B-cell_line
positive	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
as	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
nonmonocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
HeLa	NN	O	B-cell_line
and	NN	O	O
REX	NN	O	B-cell_line
.	NN	O	O

Four	NN	O	O
regions	NN	O	O
in	NN	O	O
this	NN	O	O
DNA	NN	O	B-DNA
fragment	NN	O	I-DNA
interact	NN	O	O
with	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
isolated	NN	O	O
from	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
Sp1	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
three	NN	O	O
different	NN	O	O
regions	NN	O	O
in	NN	O	O
the	NN	O	O
CD14	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
Sp1	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
-110	NN	O	B-DNA
bp	NN	O	I-DNA
)	NN	O	O
decreased	NN	O	O
tissue-specific	NN	O	O
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
transactivation	NN	O	O
experiments	NN	O	O
,	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Sp1	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
CD14	NN	O	B-protein
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CD14	NN	O	B-protein
Sp1	NN	O	B-protein
site	NN	O	O
oligonucleotides	NN	O	O
bound	NN	O	O
preferentially	NN	O	O
to	NN	O	O
a	NN	O	O
105-kDa	NN	O	B-protein
Sp1	NN	O	I-protein
species	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
higher	NN	O	O
relative	NN	O	O
levels	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
than	NN	O	O
non-monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
modification	NN	O	O
of	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
may	NN	O	O
explain	NN	O	O
how	NN	O	O
the	NN	O	O
Sp1	NN	O	B-protein
site	NN	O	O
mediates	NN	O	O
monocytic	NN	O	O
specific	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
interleukin-8	NN	O	B-DNA
AP-1	NN	O	I-DNA
and	NN	O	I-DNA
kappa	NN	O	I-DNA
B-like	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
genetic	NN	O	O
end	NN	O	O
targets	NN	O	O
of	NN	O	O
FK506-sensitive	NN	O	O
pathway	NN	O	O
accompanied	NN	O	O
by	NN	O	O
calcium	NN	O	O
mobilization	NN	O	O
.	NN	O	O

FK506	NN	O	O
,	NN	O	O
an	NN	O	O
immunosuppressant	NN	O	O
,	NN	O	O
inhibits	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
several	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
FK506	NN	O	O
suppressed	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
chemotactic	NN	O	B-protein
cytokine	NN	O	I-protein
,	NN	O	O
interleukin-8	NN	O	B-protein
(	NN	O	O
IL-8	NN	O	B-protein
)	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
activated	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
calcium	NN	O	O
(	NN	O	O
Ca2+	NN	O	O
)	NN	O	O
ionophore	NN	O	O
(	NN	O	O
ionomycin	NN	O	O
)	NN	O	O
.	NN	O	O

By	NN	O	O
deleted	NN	O	O
and	NN	O	O
mutated	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
IL-8	NN	O	B-DNA
promoters	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
and	NN	O	I-DNA
kappa	NN	O	I-DNA
B-like	NN	O	I-DNA
sites	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
responsive	NN	O	O
elements	NN	O	O
for	NN	O	O
PMA	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
.	NN	O	O

FK506	NN	O	O
suppressed	NN	O	O
the	NN	O	O
transcriptions	NN	O	O
through	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
or	NN	O	O
kappa	NN	O	B-DNA
B-like	NN	O	I-DNA
sites	NN	O	I-DNA
induced	NN	O	O
by	NN	O	O
PMA	NN	O	O
plus	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-mobilizing	NN	O	O
agents	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
those	NN	O	O
induced	NN	O	O
by	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-independent	NN	O	O
stimuli	NN	O	O
.	NN	O	O

In	NN	O	O
gel	NN	O	O
retardation	NN	O	O
analysis	NN	O	O
,	NN	O	O
FK506	NN	O	O
had	NN	O	O
little	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
of	NN	O	O
PMA/ionomycin-induced	NN	O	O
nuclear	NN	O	O
factors	NN	O	O
,	NN	O	O
which	NN	O	O
were	NN	O	O
recognized	NN	O	O
with	NN	O	O
anti-JunD	NN	O	B-protein
or	NN	O	O
c-Fos	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
FK506	NN	O	O
or	NN	O	O
EGTA	NN	O	O
(	NN	O	O
Ca2+	NN	O	O
chelator	NN	O	O
)	NN	O	O
similarly	NN	O	O
affected	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
kappa	NN	O	B-protein
B-like	NN	O	I-protein
site	NN	O	I-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
,	NN	O	O
which	NN	O	O
were	NN	O	O
not	NN	O	O
recognized	NN	O	O
by	NN	O	O
any	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
the	NN	O	O
human	NN	O	B-protein
Rel	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p49	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
confirmed	NN	O	O
the	NN	O	O
previous	NN	O	O
report	NN	O	O
that	NN	O	O
FK506	NN	O	O
suppressed	NN	O	O
the	NN	O	O
PMA/ionomycin-induced	NN	O	O
activation	NN	O	O
through	NN	O	O
authentic	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
immunoglobulin	NN	O	B-DNA
(	NN	O	I-DNA
Ig	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
to	NN	O	O
which	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
was	NN	O	O
also	NN	O	O
decreased	NN	O	O
by	NN	O	O
FK506	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
both	NN	O	O
IL-8	NN	O	B-DNA
kappa	NN	O	I-DNA
B-like	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
Ig	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
are	NN	O	O
FK506-sensitive	NN	O	O
in	NN	O	O
spite	NN	O	O
of	NN	O	O
the	NN	O	O
difference	NN	O	O
of	NN	O	O
binding	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
not	NN	O	O
only	NN	O	O
the	NN	O	O
reported	NN	O	O
IL-2	NN	O	B-DNA
NF-AT	NN	O	I-DNA
and	NN	O	I-DNA
NFIL-2A	NN	O	I-DNA
sites	NN	O	I-DNA
and	NN	O	O
Ig	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
also	NN	O	O
the	NN	O	O
IL-8	NN	O	B-protein
AP-1	NN	O	B-DNA
and	NN	O	O
kappa	NN	O	B-DNA
B-like	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
terminals	NN	O	O
of	NN	O	O
FK506-sensitive	NN	O	O
pathway	NN	O	O
involving	NN	O	O
Ca2+	NN	O	O
mobilization	NN	O	O
.	NN	O	O

-DOCSTART-	O

Androgen	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
human	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
of	NN	O	O
healthy	NN	O	O
males	NN	O	O
and	NN	O	O
females	NN	O	O
.	NN	O	O

Androgen	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
circulating	NN	O	B-cell_type
human	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
of	NN	O	O
healthy	NN	O	O
donors	NN	O	O
of	NN	O	O
both	NN	O	O
sexes	NN	O	O
.	NN	O	O

Cells	NN	O	O
were	NN	O	O
separated	NN	O	O
from	NN	O	O
blood	NN	O	O
samples	NN	O	O
on	NN	O	O
a	NN	O	O
Ficoll	NN	O	O
gradient	NN	O	O
and	NN	O	O
incubated	NN	O	O
with	NN	O	O
different	NN	O	O
concentrations	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
testosterone	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
400-fold	NN	O	O
excess	NN	O	O
of	NN	O	O
unlabelled	NN	O	O
testosterone	NN	O	O
.	NN	O	O

Binding	NN	O	O
data	NN	O	O
were	NN	O	O
derived	NN	O	O
from	NN	O	O
Scatchard	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	B-protein
sites	NN	O	I-protein
fulfil	NN	O	O
the	NN	O	O
required	NN	O	O
criteria	NN	O	O
for	NN	O	O
specific	NN	O	O
steroid	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
however	NN	O	O
differ	NN	O	O
somewhat	NN	O	O
from	NN	O	O
the	NN	O	O
classic	NN	O	O
androgen	NN	O	B-protein
receptors	NN	O	I-protein
from	NN	O	O
genital	NN	O	B-cell_type
skin	NN	O	I-cell_type
fibroblast	NN	O	I-cell_type
:	NN	O	O
in	NN	O	O
fertile	NN	O	O
adult	NN	O	O
males	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
20	NN	O	O
)	NN	O	O
the	NN	O	O
binding	NN	O	B-protein
sites	NN	O	I-protein
showed	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
a	NN	O	O
high	NN	O	O
affinity	NN	O	O
for	NN	O	O
testosterone	NN	O	O
(	NN	O	O
1.32	NN	O	O
+/-	NN	O	O
0.49	NN	O	O
nM	NN	O	O
;	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SD	NN	O	O
)	NN	O	O
,	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
a	NN	O	O
saturable	NN	O	O
capacity	NN	O	O
(	NN	O	O
184	NN	O	O
+/-	NN	O	O
52	NN	O	O
binding	NN	O	B-protein
sites	NN	O	I-protein
per	NN	O	O
cell	NN	O	O
;	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SD	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
a	NN	O	O
characteristic	NN	O	O
competitive	NN	O	O
binding	NN	O	O
profile	NN	O	O
for	NN	O	O
other	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
(	NN	O	O
relative	NN	O	O
binding	NN	O	O
affinities	NN	O	O
:	NN	O	O
testosterone	NN	O	O
=	NN	O	O
dihydrotestosterone	NN	O	O
>	NN	O	O
17	NN	O	O
beta-estradiol	NN	O	O
>	NN	O	O
progesterone	NN	O	O
,	NN	O	O
whereas	NN	O	O
aldosterone	NN	O	O
,	NN	O	O
17-hydroxy-progesterone	NN	O	O
and	NN	O	O
cortisol	NN	O	O
did	NN	O	O
not	NN	O	O
compete	NN	O	O
appreciably	NN	O	O
)	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
binding	NN	O	B-protein
sites	NN	O	I-protein
determined	NN	O	O
using	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dihydrotestosterone	NN	O	O
,	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
RU-1881	NN	O	O
,	NN	O	O
or	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
testosterone	NN	O	O
were	NN	O	O
comparable	NN	O	O
.	NN	O	O

This	NN	O	O
raises	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
androgen	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
differ	NN	O	O
from	NN	O	O
those	NN	O	O
in	NN	O	O
genital	NN	O	B-cell_type
skin	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
apparent	NN	O	O
correlation	NN	O	O
between	NN	O	O
serum	NN	O	O
testosterone	NN	O	O
concentrations	NN	O	O
and	NN	O	O
androgen	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
.	NN	O	O

In	NN	O	O
fertile	NN	O	O
women	NN	O	O
remarkable	NN	O	O
changes	NN	O	O
in	NN	O	O
androgen	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
were	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
the	NN	O	O
menstrual	NN	O	O
cycle	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
immediate	NN	O	O
preovulatory	NN	O	O
period	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
androgen	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
provides	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
the	NN	O	O
experimental	NN	O	O
basis	NN	O	O
for	NN	O	O
an	NN	O	O
hypothesis	NN	O	O
of	NN	O	O
direct	NN	O	O
,	NN	O	O
receptor-mediated	NN	O	O
effects	NN	O	O
of	NN	O	O
androgens	NN	O	O
on	NN	O	O
mature	NN	O	B-cell_type
immunocompetent	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
immunological	NN	O	O
implications	NN	O	O
of	NN	O	O
these	NN	O	O
results	NN	O	O
are	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
uses	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
costimulatory	NN	O	O
pathway	NN	O	O
.	NN	O	O

IL-8	NN	O	B-protein
,	NN	O	O
a	NN	O	O
potent	NN	O	O
chemotactic	NN	O	B-protein
factor	NN	O	I-protein
for	NN	O	O
neutrophil	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
and	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
is	NN	O	O
a	NN	O	O
proinflammatory	NN	O	O
cytokine	NN	O	O
secreted	NN	O	O
by	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
including	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
molecule	NN	O	I-protein
delivers	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
essential	NN	O	O
for	NN	O	O
lymphokine	NN	O	B-protein
production	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
a	NN	O	O
conserved	NN	O	B-DNA
sequence	NN	O	I-DNA
element	NN	O	I-DNA
found	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
several	NN	O	O
lymphokine	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Anti-CD28-stimulated	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
produced	NN	O	O
significant	NN	O	O
amounts	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
;	NN	O	O
additionally	NN	O	O
,	NN	O	O
costimulation	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
and	NN	O	O
anti-CD28	NN	O	B-protein
Abs	NN	O	I-protein
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
synergistic	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

Sequence	NN	O	O
homology	NN	O	O
,	NN	O	O
single	NN	O	O
nucleotide	NN	O	O
mutations	NN	O	O
,	NN	O	O
and	NN	O	O
anti-CD28	NN	O	B-protein
Ab	NN	O	I-protein
stimulation	NN	O	O
studies	NN	O	O
established	NN	O	O
that	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B-like	NN	O	I-DNA
sequence	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
IL-8	NN	O	B-DNA
gene	NN	O	I-DNA
functioned	NN	O	O
as	NN	O	O
a	NN	O	O
CD28	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
rapamycin	NN	O	O
,	NN	O	O
blocked	NN	O	O
the	NN	O	O
synergistic	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
expression	NN	O	O
achieved	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
and	NN	O	O
anti-	NN	O	O
CD28	NN	O	B-protein
costimulation	NN	O	O
.	NN	O	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
a	NN	O	O
CD28	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
provide	NN	O	O
new	NN	O	O
insights	NN	O	O
into	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
and	NN	O	O
persistence	NN	O	O
of	NN	O	O
immune	NN	O	O
disorders	NN	O	O
characterized	NN	O	O
by	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
psoriasis	NN	O	O
and	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
.	NN	O	O

-DOCSTART-	O

MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
signaling	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_type
activation	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

The	NN	O	O
cognate	NN	O	O
interaction	NN	O	O
between	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
APCs	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
mediated	NN	O	O
by	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	I-protein
MHC	NN	O	I-protein
)	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
delivery	NN	O	O
of	NN	O	O
activation	NN	O	O
signals	NN	O	O
to	NN	O	O
the	NN	O	O
APC	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
signals	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
co-stimulatory	NN	O	O
activity	NN	O	O
by	NN	O	O
APCs	NN	O	B-cell_type
and	NN	O	O
have	NN	O	O
important	NN	O	O
consequences	NN	O	O
for	NN	O	O
cell	NN	O	O
effector	NN	O	O
function	NN	O	O
.	NN	O	O

MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	O
also	NN	O	O
serve	NN	O	O
as	NN	O	O
receptors	NN	O	O
for	NN	O	O
B-cell	NN	O	B-cell_type
stimulation	NN	O	O
by	NN	O	O
microbial	NN	O	B-protein
superantigens	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
,	NN	O	O
Paul	NN	O	O
Scholl	NN	O	O
and	NN	O	O
Raif	NN	O	O
Geha	NN	O	O
discuss	NN	O	O
recent	NN	O	O
advances	NN	O	O
in	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
signaling	NN	O	O
and	NN	O	O
analyse	NN	O	O
their	NN	O	O
role	NN	O	O
in	NN	O	O
human	NN	O	O
B-cell	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
.	NN	O	O

Diminished	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
do	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
have	NN	O	O
diminished	NN	O	O
receptor	NN	O	O
density	NN	O	O
;	NN	O	O
thus	NN	O	O
,	NN	O	O
patients	NN	O	O
with	NN	O	O
RA	NN	O	O
should	NN	O	O
show	NN	O	O
partial	NN	O	O
resistance	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
in	NN	O	O
RA	NN	O	O
patients	NN	O	O
compared	NN	O	O
with	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
.	NN	O	O

METHODS	NN	O	O
.	NN	O	O

We	NN	O	O
determined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
on	NN	O	O
lymphocyte	NN	O	B-cell_type
proliferation	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
release	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
.	NN	O	O

Proliferation	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
release	NN	O	O
were	NN	O	O
inhibited	NN	O	O
in	NN	O	O
RA	NN	O	O
patients	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
extent	NN	O	O
as	NN	O	O
in	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
.	NN	O	O

Diminished	NN	O	O
receptor	NN	O	O
density	NN	O	O
in	NN	O	O
RA	NN	O	O
patients	NN	O	O
does	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
.	NN	O	O

-DOCSTART-	O

Arrested	NN	O	O
development	NN	O	O
:	NN	O	O
understanding	NN	O	O
v-abl	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
activity	NN	O	O
of	NN	O	O
the	NN	O	O
v-abl	NN	O	B-DNA
oncogene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
subvert	NN	O	O
the	NN	O	O
normal	NN	O	O
second	NN	O	O
messenger	NN	O	O
systems	NN	O	O
used	NN	O	O
by	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Transformation	NN	O	O
of	NN	O	O
bone	NN	O	O
marrow	NN	O	O
with	NN	O	O
the	NN	O	O
Abelson	NN	O	O
murine	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
arrested	NN	O	O
at	NN	O	O
the	NN	O	O
pre-B	NN	O	O
cell	NN	O	O
stage	NN	O	O
.	NN	O	O

Recent	NN	O	O
reports	NN	O	O
have	NN	O	O
characterized	NN	O	O
these	NN	O	O
cells	NN	O	O
expressing	NN	O	O
high	NN	O	O
v-abl	NN	O	B-DNA
kinase	NN	O	O
activity	NN	O	O
as	NN	O	O
deficient	NN	O	O
in	NN	O	O
detectable	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
low	NN	O	O
level	NN	O	O
RAG	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
v-abl	NN	O	B-DNA
may	NN	O	O
be	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
blocking	NN	O	O
these	NN	O	O
two	NN	O	O
crucial	NN	O	O
elements	NN	O	O
in	NN	O	O
the	NN	O	O
maturation	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
:	NN	O	O
a	NN	O	O
possible	NN	O	O
marker	NN	O	O
of	NN	O	O
brain	NN	O	O
involution	NN	O	O
?	NN	O	O

A	NN	O	O
similarity	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
found	NN	O	O
between	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
brain	NN	O	O
and	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
tissue	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
as	NN	O	O
a	NN	O	O
possible	NN	O	O
marker	NN	O	O
of	NN	O	O
brain	NN	O	O
involution	NN	O	O
.	NN	O	O

Type	NN	O	B-protein
I	NN	O	I-protein
corticosteroid	NN	O	I-protein
receptors	NN	O	I-protein
are	NN	O	O
down	NN	O	O
regulated	NN	O	O
by	NN	O	O
excess	NN	O	O
of	NN	O	O
mineralocorticoids	NN	O	O
(	NN	O	O
primary	NN	O	O
and	NN	O	O
secondary	NN	O	O
hyperaldosteronism	NN	O	O
,	NN	O	O
pseudohyperaldosteronism	NN	O	O
)	NN	O	O
and	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
(	NN	O	O
Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
)	NN	O	O
.	NN	O	O
Type	NN	O	B-protein
II	NN	O	I-protein
corticosteroid	NN	O	I-protein
receptors	NN	O	I-protein
are	NN	O	O
not	NN	O	O
reduced	NN	O	O
by	NN	O	O
excess	NN	O	O
of	NN	O	O
endogenous	NN	O	O
corticosteroids	NN	O	O
(	NN	O	O
Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
normal	NN	O	O
adults	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
direct	NN	O	O
significant	NN	O	O
correlation	NN	O	O
between	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
and	NN	O	O
Type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	O
between	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
and	NN	O	O
Type	NN	O	B-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
in	NN	O	O
Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
the	NN	O	O
correlation	NN	O	O
is	NN	O	O
inverse	NN	O	O
between	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
at	NN	O	O
8	NN	O	O
a.m.	NN	O	O
and	NN	O	O
Type	NN	O	B-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
an	NN	O	O
aged	NN	O	O
population	NN	O	O
the	NN	O	O
mean	NN	O	O
numbers	NN	O	O
of	NN	O	O
Type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	O
of	NN	O	O
Type	NN	O	B-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
are	NN	O	O
lower	NN	O	O
and	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
is	NN	O	O
higher	NN	O	O
than	NN	O	O
in	NN	O	O
adult	NN	O	O
controls	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
increase	NN	O	O
of	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
is	NN	O	O
not	NN	O	O
followed	NN	O	O
by	NN	O	O
a	NN	O	O
clinical	NN	O	O
picture	NN	O	O
of	NN	O	O
hypercorticism	NN	O	O
.	NN	O	O

Corticosteroid	NN	O	B-protein
Type	NN	O	I-protein
I	NN	O	I-protein
and	NN	O	I-protein
Type	NN	O	I-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
are	NN	O	O
inversely	NN	O	O
correlated	NN	O	O
with	NN	O	O
age	NN	O	O
.	NN	O	O

After	NN	O	O
dexamethasone	NN	O	O
suppression	NN	O	O
(	NN	O	O
1	NN	O	O
mg	NN	O	O
at	NN	O	O
11	NN	O	O
p.m.	NN	O	O
)	NN	O	O
Type	NN	O	B-protein
I	NN	O	I-protein
receptors	NN	O	I-protein
always	NN	O	O
decrease	NN	O	O
in	NN	O	O
controls	NN	O	O
while	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
Type	NN	O	B-protein
II	NN	O	I-protein
is	NN	O	O
not	NN	O	O
homogeneous	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
aged	NN	O	O
group	NN	O	O
of	NN	O	O
patients	NN	O	O
,	NN	O	O
both	NN	O	O
receptors	NN	O	O
are	NN	O	O
reduced	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
decrease	NN	O	O
with	NN	O	O
age	NN	O	O
of	NN	O	O
corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
is	NN	O	O
possibly	NN	O	O
related	NN	O	O
to	NN	O	O
a	NN	O	O
physiological	NN	O	O
involution	NN	O	O
of	NN	O	O
corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
that	NN	O	O
this	NN	O	O
reduction	NN	O	O
does	NN	O	O
increase	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentration	NN	O	O
,	NN	O	O
without	NN	O	O
affecting	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
effector	NN	O	O
mechanism	NN	O	O
.	NN	O	O

-DOCSTART-	O

All-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
and	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
co-operate	NN	O	O
to	NN	O	O
promote	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
promyeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
HL60	NN	O	B-cell_line
to	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

A	NN	O	O
basis	NN	O	O
for	NN	O	O
differentiation	NN	O	O
therapy	NN	O	O
of	NN	O	O
leukemias	NN	O	O
is	NN	O	O
provided	NN	O	O
by	NN	O	O
knowledge	NN	O	O
of	NN	O	O
agents	NN	O	O
which	NN	O	O
induce	NN	O	O
specific	NN	O	O
lineage	NN	O	O
maturation	NN	O	O
.	NN	O	O

All-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
induces	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
is	NN	O	O
used	NN	O	O
to	NN	O	O
treat	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
RA	NN	O	O
did	NN	O	O
not	NN	O	O
induced	NN	O	O
neutrophil	NN	O	O
differentiation	NN	O	O
in	NN	O	O
serum-free	NN	O	B-cell_line
grown	NN	O	I-cell_line
HL60	NN	O	I-cell_line
cells	NN	O	I-cell_line
whereas	NN	O	O
50	NN	O	O
nM	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
D3	NN	O	O
)	NN	O	O
induced	NN	O	O
maximal	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Increasing	NN	O	O
RA	NN	O	O
concentrations	NN	O	O
reduced	NN	O	O
the	NN	O	O
D3	NN	O	O
concentration	NN	O	O
required	NN	O	O
for	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Cells	NN	O	O
treated	NN	O	O
with	NN	O	O
5	NN	O	O
nM	NN	O	O
D3	NN	O	O
showed	NN	O	O
little	NN	O	O
response	NN	O	O
,	NN	O	O
but	NN	O	O
differentiated	NN	O	O
maximally	NN	O	O
with	NN	O	O
5	NN	O	O
nM	NN	O	O
D3	NN	O	O
and	NN	O	O
10	NN	O	O
nM	NN	O	O
RA	NN	O	O
.	NN	O	O

The	NN	O	O
D3	NN	O	O
analogs	NN	O	O
MC903	NN	O	O
,	NN	O	O
EB1089	NN	O	O
and	NN	O	O
KH1060	NN	O	O
were	NN	O	O
more	NN	O	O
potent	NN	O	O
inducers	NN	O	O
of	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
extent	NN	O	O
to	NN	O	O
which	NN	O	O
analog	NN	O	O
activity	NN	O	O
was	NN	O	O
increased	NN	O	O
after	NN	O	O
cotreatment	NN	O	O
with	NN	O	O
RA	NN	O	O
was	NN	O	O
inversely	NN	O	O
related	NN	O	O
to	NN	O	O
potency	NN	O	O
.	NN	O	O

Twenty-four	NN	O	O
hour	NN	O	O
treatment	NN	O	O
with	NN	O	O
10	NN	O	O
nM	NN	O	O
RA	NN	O	B-cell_line
primed	NN	O	I-cell_line
cells	NN	O	I-cell_line
for	NN	O	O
response	NN	O	O
to	NN	O	O
5	NN	O	O
nM	NN	O	O
D3	NN	O	O
;	NN	O	O
the	NN	O	O
reverse	NN	O	O
sequence	NN	O	O
being	NN	O	O
ineffective	NN	O	O
.	NN	O	O

Priming	NN	O	O
with	NN	O	O
10	NN	O	O
nM	NN	O	O
RA	NN	O	O
,	NN	O	O
or	NN	O	O
subsequent	NN	O	O
treatment	NN	O	O
with	NN	O	O
D3	NN	O	O
(	NN	O	O
5	NN	O	O
nM	NN	O	O
)	NN	O	O
,	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
expression	NN	O	O
of	NN	O	O
mRNAs	NN	O	B-RNA
encoding	NN	O	O
receptors	NN	O	B-protein
for	NN	O	O
D3	NN	O	O
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
RA	NN	O	O
(	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
or	NN	O	O
9-CIS	NN	O	O
RA	NN	O	O
(	NN	O	O
RXR	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	I-protein
beta	NN	O	I-protein
,	NN	O	I-protein
gamma	NN	O	I-protein
)	NN	O	O
.	NN	O	O

That	NN	O	O
RA	NN	O	O
promotes	NN	O	O
both	NN	O	O
neutrophil	NN	O	O
and	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
has	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
RA	NN	O	O
and	NN	O	O
D3	NN	O	O
in	NN	O	O
treatment	NN	O	O
of	NN	O	O
leukemias	NN	O	O
and	NN	O	O
provides	NN	O	O
insight	NN	O	O
into	NN	O	O
mechanisms	NN	O	O
whereby	NN	O	O
RAR	NN	O	O
,	NN	O	O
VDR	NN	O	B-protein
and	NN	O	O
RXR	NN	O	B-protein
facilitate	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
An	NN	O	O
overexpression	NN	O	O
of	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
blocks	NN	O	O
myeloid	NN	O	B-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
at	NN	O	O
the	NN	O	O
promyelocyte	NN	O	O
stage	NN	O	O
]	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
vitamin	NN	O	O
A	NN	O	O
derivative	NN	O	O
,	NN	O	O
exerts	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
biological	NN	O	O
effects	NN	O	O
related	NN	O	O
to	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
pleiotropic	NN	O	O
effects	NN	O	O
of	NN	O	O
RA	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
through	NN	O	O
specific	NN	O	O
nuclear	NN	O	O
RA	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
RARs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

RARs	NN	O	B-protein
are	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
steroid/thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
and	NN	O	O
exhibit	NN	O	O
a	NN	O	O
molecular	NN	O	O
structure	NN	O	O
that	NN	O	O
possess	NN	O	O
discrete	NN	O	O
DNA-binding	NN	O	O
and	NN	O	O
RA	NN	O	B-protein
(	NN	O	I-protein
ligand	NN	O	I-protein
)	NN	O	I-protein
-binding	NN	O	I-protein
domains	NN	O	I-protein
.	NN	O	O

In	NN	O	O
hematopoietic	NN	O	O
system	NN	O	O
,	NN	O	O
RA	NN	O	O
and	NN	O	O
RARs	NN	O	B-protein
,	NN	O	O
predominantly	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
may	NN	O	O
play	NN	O	O
key	NN	O	O
roles	NN	O	O
for	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
currently	NN	O	O
unknown	NN	O	O
how	NN	O	O
RA	NN	O	O
and	NN	O	O
RARs	NN	O	B-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
regulating	NN	O	O
normal	NN	O	O
hematopoietic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

To	NN	O	O
make	NN	O	O
clear	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
RA	NN	O	O
and	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
in	NN	O	O
the	NN	O	O
normal	NN	O	O
hematopoiesis	NN	O	O
,	NN	O	O
I	NN	O	O
have	NN	O	O
introduced	NN	O	O
the	NN	O	O
construct	NN	O	O
of	NN	O	O
human	NN	O	B-protein
RAR	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
hRAR	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
into	NN	O	O
murine	NN	O	B-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
retroviral	NN	O	O
vector	NN	O	O
,	NN	O	O
and	NN	O	O
selected	NN	O	O
infected	NN	O	O
cells	NN	O	O
with	NN	O	O
drug	NN	O	B-DNA
resistant	NN	O	I-DNA
marker	NN	O	I-DNA
(	NN	O	O
Neo	NN	O	B-DNA
(	NN	O	I-DNA
r	NN	O	I-DNA
)	NN	O	I-DNA
)	NN	O	O
cultured	NN	O	O
on	NN	O	O
the	NN	O	O
stroma	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
PA6-neo	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
and	NN	O	O
analyzed	NN	O	O
the	NN	O	O
behavior	NN	O	O
of	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

All	NN	O	O
of	NN	O	O
procedure	NN	O	O
were	NN	O	O
done	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Most	NN	O	O
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
hRAR	NN	O	B-protein
alpha	NN	O	I-protein
exhibited	NN	O	O
promyelocytic	NN	O	B-cell_type
morphology	NN	O	I-cell_type
and	NN	O	O
were	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
blocked	NN	O	O
at	NN	O	O
the	NN	O	O
promyelocytic	NN	O	O
stage	NN	O	O
in	NN	O	O
their	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
these	NN	O	O
immature	NN	O	B-cell_type
cells	NN	O	I-cell_type
differentiated	NN	O	O
terminally	NN	O	O
into	NN	O	O
mature	NN	O	O
granulocytes	NN	O	O
by	NN	O	O
adding	NN	O	O
with	NN	O	O
RA	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
.	NN	O	O

RAR	NN	O	B-protein
alpha	NN	O	I-protein
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
also	NN	O	O
able	NN	O	O
to	NN	O	O
differentiate	NN	O	O
into	NN	O	O
mature	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
both	NN	O	O
of	NN	O	O
long	NN	O	O
term	NN	O	O
culture	NN	O	O
and	NN	O	O
IL3	NN	O	B-cell_line
colony	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
an	NN	O	O
overexpression	NN	O	O
of	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
alone	NN	O	O
is	NN	O	O
effective	NN	O	O
to	NN	O	O
suppress	NN	O	O
myeloid	NN	O	B-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
and	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
of	NN	O	O
myeloid	NN	O	O
precursors	NN	O	O
.	NN	O	O

The	NN	O	O
system	NN	O	O
described	NN	O	O
here	NN	O	O
may	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
studying	NN	O	O
the	NN	O	O
the	NN	O	O
essential	NN	O	B-DNA
genes	NN	O	I-DNA
for	NN	O	O
differentiation	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Hypoxic	NN	O	O
induction	NN	O	O
of	NN	O	O
interleukin-8	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Because	NN	O	O
leukocyte-mediated	NN	O	O
tissue	NN	O	O
damage	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
pathologic	NN	O	O
picture	NN	O	O
in	NN	O	O
ischemia/reperfusion	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
sought	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
PMNs	NN	O	B-cell_type
are	NN	O	O
directed	NN	O	O
into	NN	O	O
hypoxic	NN	O	O
tissue	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
ECs	NN	O	B-cell_type
)	NN	O	O
in	NN	O	O
hypoxia	NN	O	O
,	NN	O	O
PO2	NN	O	O
approximately	NN	O	O
14-18	NN	O	O
Torr	NN	O	O
,	NN	O	O
led	NN	O	O
to	NN	O	O
time-dependent	NN	O	O
release	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
antigen	NN	O	O
into	NN	O	O
the	NN	O	O
conditioned	NN	O	O
medium	NN	O	O
;	NN	O	O
this	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
increased	NN	O	O
chemotactic	NN	O	O
activity	NN	O	O
for	NN	O	O
PMNs	NN	O	B-cell_type
,	NN	O	O
blocked	NN	O	O
by	NN	O	O
antibody	NN	O	O
to	NN	O	O
IL-8	NN	O	B-protein
.	NN	O	O

Production	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
by	NN	O	O
hypoxic	NN	O	O
ECs	NN	O	B-cell_type
occurred	NN	O	O
concomitantly	NN	O	O
with	NN	O	O
both	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-8	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
based	NN	O	O
on	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
analysis	NN	O	O
,	NN	O	O
and	NN	O	O
increased	NN	O	O
IL-8	NN	O	B-protein
transcription	NN	O	O
,	NN	O	O
based	NN	O	O
on	NN	O	O
nuclear	NN	O	O
run-on	NN	O	O
assays	NN	O	O
.	NN	O	O

Northern	NN	O	O
analysis	NN	O	O
of	NN	O	O
mRNA	NN	O	O
from	NN	O	O
hypoxic	NN	O	O
ECs	NN	O	B-cell_type
also	NN	O	O
demonstrated	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
macrophage	NN	O	B-protein
chemotactic	NN	O	I-protein
protein-1	NN	O	I-protein
,	NN	O	O
another	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
chemokine	NN	O	B-protein
superfamily	NN	O	I-protein
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

IL-8	NN	O	B-DNA
gene	NN	O	I-DNA
induction	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
increased	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
hypoxic	NN	O	B-cell_type
ECs	NN	O	I-cell_type
for	NN	O	O
the	NN	O	O
NF-kB	NN	O	O
site	NN	O	O
.	NN	O	O

Studies	NN	O	O
with	NN	O	O
human	NN	O	O
umbilical	NN	O	O
vein	NN	O	O
segments	NN	O	O
exposed	NN	O	O
to	NN	O	O
hypoxia	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
increased	NN	O	O
elaboration	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
antigen	NN	O	I-protein
compared	NN	O	O
with	NN	O	O
normoxic	NN	O	O
controls	NN	O	O
.	NN	O	O

In	NN	O	O
mice	NN	O	O
exposed	NN	O	O
to	NN	O	O
hypoxia	NN	O	O
(	NN	O	O
PO2	NN	O	O
approximately	NN	O	O
30-40	NN	O	O
Torr	NN	O	O
)	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
increased	NN	O	O
pulmonary	NN	O	O
leukostasis	NN	O	O
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
increased	NN	O	O
myeloperoxidase	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
tissue	NN	O	O
homogenates	NN	O	O
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
,	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
transcripts	NN	O	O
for	NN	O	O
IP-10	NN	O	B-protein
,	NN	O	O
a	NN	O	O
murine	NN	O	O
homologue	NN	O	O
in	NN	O	O
the	NN	O	O
chemokine	NN	O	B-protein
family	NN	O	I-protein
related	NN	O	O
to	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
hypoxic	NN	O	O
lung	NN	O	O
tissue	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
hypoxia	NN	O	O
constitutes	NN	O	O
a	NN	O	O
stimulus	NN	O	O
for	NN	O	O
leukocyte	NN	O	O
chemotaxis	NN	O	O
and	NN	O	O
tissue	NN	O	O
leukostasis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Thrombin	NN	O	B-protein
and	NN	O	O
thrombin	NN	O	B-protein
receptor	NN	O	I-protein
agonist	NN	O	O
peptide	NN	O	O
induce	NN	O	O
early	NN	O	O
events	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
synergize	NN	O	O
with	NN	O	O
TCR	NN	O	B-protein
cross-linking	NN	O	O
for	NN	O	O
CD69	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
interleukin	NN	O	O
2	NN	O	O
production	NN	O	O
.	NN	O	O

Thrombin	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
induced	NN	O	O
a	NN	O	O
transient	NN	O	O
increase	NN	O	O
in	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
.	NN	O	O

Proteolytic	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	B-protein
was	NN	O	O
required	NN	O	O
for	NN	O	O
this	NN	O	O
effect	NN	O	O
since	NN	O	O
diisopropyl	NN	O	B-protein
fluorophosphate-thrombin	NN	O	I-protein
failed	NN	O	O
to	NN	O	O
increase	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
hirudin	NN	O	O
and	NN	O	O
anti-thrombin	NN	O	B-protein
III	NN	O	I-protein
inhibited	NN	O	O
the	NN	O	O
thrombin-induced	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
rise	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
synthetic	NN	O	O
thrombin	NN	O	B-protein
receptor	NN	O	I-protein
agonist	NN	O	O
peptide	NN	O	O
(	NN	O	O
TRP	NN	O	O
)	NN	O	O
of	NN	O	O
7	NN	O	O
residues	NN	O	O
(	NN	O	O
SFLLRNP	NN	O	O
)	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
as	NN	O	O
effective	NN	O	O
as	NN	O	O
thrombin	NN	O	B-protein
for	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
mobilization	NN	O	O
,	NN	O	O
and	NN	O	O
both	NN	O	O
agonists	NN	O	O
induced	NN	O	O
Ca2+	NN	O	O
release	NN	O	O
exclusively	NN	O	O
from	NN	O	O
internal	NN	O	O
stores	NN	O	O
.	NN	O	O

Thrombin	NN	O	B-protein
stimulated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
several	NN	O	O
proteins	NN	O	O
of	NN	O	O
molecular	NN	O	B-protein
mass	NN	O	I-protein
40	NN	O	I-protein
,	NN	O	I-protein
42	NN	O	I-protein
,	NN	O	I-protein
70	NN	O	I-protein
,	NN	O	I-protein
120	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
130	NN	O	I-protein
kDa	NN	O	I-protein
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
good	NN	O	O
correlation	NN	O	O
between	NN	O	O
thrombin	NN	O	B-protein
-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
latter	NN	O	O
three	NN	O	O
proteins	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
mobilization	NN	O	O
.	NN	O	O

Thrombin	NN	O	B-protein
and	NN	O	O
TRP	NN	O	O
also	NN	O	O
caused	NN	O	O
translocation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
from	NN	O	O
the	NN	O	O
cytosol	NN	O	O
to	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
likely	NN	O	O
consequence	NN	O	O
of	NN	O	O
these	NN	O	O
events	NN	O	O
,	NN	O	O
thrombin	NN	O	B-protein
activated	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
NF-kB	NN	O	I-protein
.	NN	O	O

Several	NN	O	O
cell	NN	O	O
lines	NN	O	O
of	NN	O	O
hematopoietic	NN	O	O
origin	NN	O	O
including	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
HPB.ALL	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
erythroleukemic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
K562	NN	O	I-cell_line
were	NN	O	O
responsive	NN	O	O
to	NN	O	O
thrombin	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
others	NN	O	O
such	NN	O	O
as	NN	O	O
THP1	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
myelomonocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
BL2	NN	O	B-cell_line
,	NN	O	I-cell_line
a	NN	O	O
Burkitt	NN	O	B-cell_line
lymphoma	NN	O	I-cell_line
were	NN	O	O
refractory	NN	O	O
to	NN	O	O
thrombin	NN	O	B-protein
or	NN	O	O
TRP	NN	O	O
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
magnitude	NN	O	O
of	NN	O	O
the	NN	O	O
thrombin	NN	O	B-protein
response	NN	O	O
in	NN	O	O
the	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
paralleled	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
thrombin	NN	O	B-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
phytohemagglutinin	NN	O	B-protein
and	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
dramatic	NN	O	O
inhibition	NN	O	O
of	NN	O	O
thrombin	NN	O	B-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
concomitant	NN	O	O
loss	NN	O	O
of	NN	O	O
the	NN	O	O
thrombin	NN	O	B-protein
response	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
thrombin	NN	O	B-protein
and	NN	O	O
TRP	NN	O	O
enhanced	NN	O	O
CD69	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
production	NN	O	O
induced	NN	O	O
by	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
cross-linking	NN	O	O
in	NN	O	O
both	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
findings	NN	O	O
highlight	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
thrombin	NN	O	B-protein
as	NN	O	O
a	NN	O	O
potential	NN	O	O
regulator	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stress	NN	O	O
response	NN	O	O
of	NN	O	O
senescent	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
reduced	NN	O	O
hsp70	NN	O	B-protein
is	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
proliferative	NN	O	O
block	NN	O	O
.	NN	O	O

Senescent	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
cultures	NN	O	I-cell_line
are	NN	O	O
unable	NN	O	O
to	NN	O	O
undergo	NN	O	O
proliferation	NN	O	O
,	NN	O	O
but	NN	O	O
show	NN	O	O
no	NN	O	O
difference	NN	O	O
from	NN	O	O
early	NN	O	B-cell_line
passage	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
cytotoxic	NN	O	O
function	NN	O	O
or	NN	O	O
surface	NN	O	O
antigenic	NN	O	O
profile	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
feature	NN	O	O
of	NN	O	O
senescent	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
the	NN	O	O
dramatic	NN	O	O
reduction	NN	O	O
in	NN	O	O
hsp70	NN	O	B-protein
production	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
heat	NN	O	O
shock	NN	O	O
.	NN	O	O

This	NN	O	O
decline	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
heat	NN	O	I-DNA
shock	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
progressive	NN	O	O
decline	NN	O	O
in	NN	O	O
the	NN	O	O
heat	NN	O	O
shock	NN	O	O
response	NN	O	O
of	NN	O	O
cultured	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
percent	NN	O	O
proliferative	NN	O	O
life	NN	O	O
span	NN	O	O
completed	NN	O	O
rather	NN	O	O
than	NN	O	O
with	NN	O	O
the	NN	O	O
actual	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
at	NN	O	O
the	NN	O	O
time	NN	O	O
of	NN	O	O
heat	NN	O	O
shock	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
for	NN	O	O
senescent	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
the	NN	O	O
reduced	NN	O	O
ability	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
heat	NN	O	O
shock	NN	O	O
by	NN	O	O
producing	NN	O	O
hsp70	NN	O	B-protein
,	NN	O	O
although	NN	O	O
possibly	NN	O	O
lying	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
,	NN	O	O
may	NN	O	O
nevertheless	NN	O	O
be	NN	O	O
unrelated	NN	O	O
to	NN	O	O
the	NN	O	O
reduced	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
or	NN	O	O
the	NN	O	O
diminished	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
also	NN	O	O
manifested	NN	O	O
by	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
phospholipase	NN	O	B-protein
D	NN	O	I-protein
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
has	NN	O	O
been	NN	O	O
ascribed	NN	O	O
to	NN	O	O
the	NN	O	O
early	NN	O	O
events	NN	O	O
leading	NN	O	O
to	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
phospholipase	NN	O	B-protein
D	NN	O	I-protein
(	NN	O	O
PLD	NN	O	B-protein
)	NN	O	O
may	NN	O	O
regulate	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
in	NN	O	O
human	NN	O	O
T	NN	O	O
cells	NN	O	O
by	NN	O	O
transfecting	NN	O	O
human	NN	O	O
T	NN	O	B-cell_line
lymphocyte	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
a	NN	O	O
plasmid	NN	O	O
containing	NN	O	O
an	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
detected	NN	O	O
activatable	NN	O	O
PLD	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
addition	NN	O	O
of	NN	O	O
phosphatidic	NN	O	O
acid	NN	O	O
(	NN	O	O
PA	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
physiologic	NN	O	O
product	NN	O	O
of	NN	O	O
PLD	NN	O	B-protein
action	NN	O	O
on	NN	O	O
phospholipids	NN	O	O
,	NN	O	O
is	NN	O	O
rapidly	NN	O	O
incorporated	NN	O	O
into	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
leads	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
activated	NN	O	O
both	NN	O	O
PLD	NN	O	B-protein
and	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
.	NN	O	O

Wortmannin	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
receptor-coupled	NN	O	O
PLD	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
blocked	NN	O	O
the	NN	O	O
anti-CD3	NN	O	B-protein
-induced	NN	O	O
increases	NN	O	O
in	NN	O	O
both	NN	O	O
PLD	NN	O	B-protein
activity	NN	O	O
and	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
a	NN	O	O
good	NN	O	O
correlation	NN	O	O
in	NN	O	O
the	NN	O	O
transfected	NN	O	B-cell_line
cells	NN	O	I-cell_line
between	NN	O	O
PLD	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
under	NN	O	O
different	NN	O	O
experimental	NN	O	O
conditions	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
ethanol	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
the	NN	O	O
PLD	NN	O	B-protein
pathway	NN	O	O
,	NN	O	O
blocked	NN	O	O
the	NN	O	O
anti-CD3-stimulated	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
anti-CD3	NN	O	B-protein
-mediated	NN	O	O
response	NN	O	O
was	NN	O	O
not	NN	O	O
inhibited	NN	O	O
by	NN	O	O
neomycin	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
phosphoinositide	NN	O	O
hydrolysis	NN	O	O
.	NN	O	O

The	NN	O	O
increases	NN	O	O
in	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
induced	NN	O	O
by	NN	O	O
PA	NN	O	O
or	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
were	NN	O	O
efficiently	NN	O	O
abrogated	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
propranolol	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
PA	NN	O	O
phosphohydrolase	NN	O	B-protein
and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
PA	NN	O	O
-and	NN	O	O
the	NN	O	O
anti-CD3	NN	O	B-protein
-induced	NN	O	O
increases	NN	O	O
in	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
were	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
inhibitors	NN	O	O
or	NN	O	O
by	NN	O	O
PKC	NN	O	B-protein
down-regulation	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
PLD	NN	O	B-protein
stimulation	NN	O	O
can	NN	O	O
activate	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
factor	NN	O	O
activity	NN	O	O
by	NN	O	O
PA	NN	O	O
is	NN	O	O
mediated	NN	O	O
via	NN	O	O
PKC	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
either	NN	O	O
through	NN	O	O
a	NN	O	O
direct	NN	O	O
activating	NN	O	O
effect	NN	O	O
of	NN	O	O
PA	NN	O	O
or	NN	O	O
through	NN	O	O
PA-derived	NN	O	O
diacylglycerol	NN	O	O
formation	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
also	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
role	NN	O	O
of	NN	O	O
PLD	NN	O	B-protein
-derived	NN	O	O
lipids	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
resulting	NN	O	O
from	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
TCR/CD3	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
increased	NN	O	O
PLD	NN	O	B-protein
activity	NN	O	O
can	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Upregulation	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-DNA
by	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
latent	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
LMP1	NN	O	B-protein
:	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	O
response	NN	O	O
that	NN	O	O
is	NN	O	O
delayed	NN	O	O
relative	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
to	NN	O	O
induction	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
markers	NN	O	I-protein
.	NN	O	O

An	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
latent	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
LMP1	NN	O	B-protein
to	NN	O	O
enhance	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
infected	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
upregulation	NN	O	O
of	NN	O	O
the	NN	O	O
bcl-2	NN	O	B-DNA
oncogene	NN	O	I-DNA
was	NN	O	O
first	NN	O	O
suggested	NN	O	O
by	NN	O	O
experiments	NN	O	O
involving	NN	O	O
gene	NN	O	O
transfection	NN	O	O
and	NN	O	O
the	NN	O	O
selection	NN	O	O
of	NN	O	O
stable	NN	O	O
LMP1+	NN	O	B-cell_line
clones	NN	O	I-cell_line
(	NN	O	O
S.Henderson	NN	O	O
,	NN	O	O
M.	NN	O	O
Rowe	NN	O	O
,	NN	O	O
C.Gregory	NN	O	O
,	NN	O	O
F.Wang	NN	O	O
,	NN	O	O
E.Kieff	NN	O	O
,	NN	O	O
and	NN	O	O
A.Rickinson	NN	O	O
,	NN	O	O
Cell	NN	O	O
65	NN	O	O
:	NN	O	O
1107-1115	NN	O	O
,	NN	O	O
1991	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
not	NN	O	O
possible	NN	O	O
to	NN	O	O
ascertain	NN	O	O
whether	NN	O	O
Bcl-2	NN	O	B-protein
upregulation	NN	O	O
was	NN	O	O
a	NN	O	O
specific	NN	O	O
consequence	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
expression	NN	O	O
or	NN	O	O
an	NN	O	O
artifact	NN	O	O
of	NN	O	O
the	NN	O	O
selection	NN	O	O
procedure	NN	O	O
whereby	NN	O	O
rare	NN	O	O
Bcl-2+	NN	O	B-cell_line
cells	NN	O	I-cell_line
already	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
starting	NN	O	O
population	NN	O	O
might	NN	O	O
best	NN	O	O
be	NN	O	O
able	NN	O	O
to	NN	O	O
tolerate	NN	O	O
the	NN	O	O
potentially	NN	O	O
toxic	NN	O	O
effects	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
reexamined	NN	O	O
this	NN	O	O
issue	NN	O	O
by	NN	O	O
using	NN	O	O
two	NN	O	O
different	NN	O	O
experimental	NN	O	O
approaches	NN	O	O
that	NN	O	O
allowed	NN	O	O
LMP1	NN	O	B-protein
-induced	NN	O	O
effects	NN	O	O
to	NN	O	O
be	NN	O	O
monitored	NN	O	O
immediately	NN	O	O
following	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	O
protein	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
selective	NN	O	O
pressures	NN	O	O
;	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
and	NN	O	O
upregulation	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	B-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
ICAM-1	NN	O	B-protein
were	NN	O	O
used	NN	O	O
as	NN	O	O
early	NN	O	O
indices	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
function	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
first	NN	O	O
approach	NN	O	O
,	NN	O	O
stable	NN	O	O
clones	NN	O	O
of	NN	O	O
two	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
carrying	NN	O	O
an	NN	O	O
LMP1	NN	O	B-protein
gene	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	O
metallothionein	NN	O	B-DNA
promoter	NN	O	I-DNA
were	NN	O	O
induced	NN	O	O
to	NN	O	O
express	NN	O	O
LMP1	NN	O	B-protein
in	NN	O	O
all	NN	O	O
cells	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NK-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
upregulation	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
occurred	NN	O	O
within	NN	O	O
24	NN	O	O
h	NN	O	O
and	NN	O	O
were	NN	O	O
followed	NN	O	O
at	NN	O	O
48	NN	O	O
to	NN	O	O
72	NN	O	O
h	NN	O	O
by	NN	O	O
upregulation	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
second	NN	O	O
approach	NN	O	O
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
generality	NN	O	O
of	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
by	NN	O	O
transiently	NN	O	O
expressing	NN	O	O
LMP1	NN	O	B-protein
from	NN	O	O
a	NN	O	O
strong	NN	O	O
constitutively	NN	O	O
active	NN	O	O
promoter	NN	O	O
in	NN	O	O
a	NN	O	O
range	NN	O	O
of	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

All	NN	O	O
six	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
tested	NN	O	O
showed	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
LMP1	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
was	NN	O	O
followed	NN	O	O
in	NN	O	O
five	NN	O	O
of	NN	O	O
six	NN	O	O
lines	NN	O	O
by	NN	O	O
expression	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
and	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
same	NN	O	O
experiments	NN	O	O
,	NN	O	O
all	NN	O	O
three	NN	O	O
non-	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
showed	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
ICAM-1	NN	O	B-protein
upregulation	NN	O	O
but	NN	O	O
never	NN	O	O
any	NN	O	O
effect	NN	O	O
upon	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
conclude	NN	O	O
that	NN	O	O
Bcl-2	NN	O	B-protein
upregulation	NN	O	O
is	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
panoply	NN	O	O
of	NN	O	O
cellular	NN	O	O
changes	NN	O	O
induced	NN	O	O
by	NN	O	O
LMP1	NN	O	B-protein
but	NN	O	O
that	NN	O	O
the	NN	O	O
effect	NN	O	O
is	NN	O	O
cell	NN	O	O
type	NN	O	O
specific	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
also	NN	O	O
suggest	NN	O	O
that	NN	O	O
whilst	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
may	NN	O	O
be	NN	O	O
an	NN	O	O
essential	NN	O	O
component	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
,	NN	O	O
other	NN	O	O
cell-specific	NN	O	B-protein
factors	NN	O	I-protein
may	NN	O	O
be	NN	O	O
required	NN	O	O
to	NN	O	O
effect	NN	O	O
some	NN	O	O
functions	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Long-term	NN	O	O
inositol	NN	O	O
phosphate	NN	O	O
release	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
correlates	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
and	NN	O	O
NF-AT	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
anti-CD3-activated	NN	O	B-cell_line
peripheral	NN	O	I-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
cascade	NN	O	O
of	NN	O	O
events	NN	O	O
within	NN	O	O
the	NN	O	O
first	NN	O	O
few	NN	O	O
minutes	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
stimulation	NN	O	O
has	NN	O	O
been	NN	O	O
well	NN	O	O
characterized	NN	O	O
.	NN	O	O

Although	NN	O	O
many	NN	O	O
second	NN	O	O
messengers	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
model	NN	O	O
systems	NN	O	O
,	NN	O	O
the	NN	O	O
rate-limiting	NN	O	O
step	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
not	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

To	NN	O	O
model	NN	O	O
effective	NN	O	O
versus	NN	O	O
ineffective	NN	O	O
CD3	NN	O	B-protein
-mediated	NN	O	O
stimulation	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
used	NN	O	O
two	NN	O	O
anti-CD3	NN	O	B-protein
mAbs	NN	O	I-protein
that	NN	O	O
differ	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
stimulate	NN	O	O
purified	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
OKT3	NN	O	B-protein
,	NN	O	O
which	NN	O	O
causes	NN	O	O
early	NN	O	O
second	NN	O	O
messenger	NN	O	O
generation	NN	O	O
but	NN	O	O
is	NN	O	O
unable	NN	O	O
to	NN	O	O
activate	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
without	NN	O	O
a	NN	O	O
second	NN	O	O
signal	NN	O	O
,	NN	O	O
and	NN	O	O
64.1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
stimulates	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
on	NN	O	O
its	NN	O	O
own	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
was	NN	O	O
similar	NN	O	O
for	NN	O	O
both	NN	O	O
mAbs	NN	O	B-protein
over	NN	O	O
a	NN	O	O
period	NN	O	O
of	NN	O	O
hours	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
inositol	NN	O	O
phosphate	NN	O	O
response	NN	O	O
was	NN	O	O
stronger	NN	O	O
for	NN	O	O
64.1	NN	O	B-protein
than	NN	O	O
for	NN	O	O
OKT3	NN	O	B-protein
.	NN	O	O

To	NN	O	O
tie	NN	O	O
these	NN	O	O
events	NN	O	O
to	NN	O	O
gene	NN	O	O
activation	NN	O	O
,	NN	O	O
we	NN	O	O
measured	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
NF-AT	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
after	NN	O	O
anti-CD3	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Both	NN	O	O
stimuli	NN	O	O
induced	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
components	NN	O	I-protein
(	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
(	NN	O	O
RelA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
p50	NN	O	B-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
)	NN	O	O
)	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
only	NN	O	O
64.1	NN	O	B-protein
induced	NN	O	O
NF-AT	NN	O	B-protein
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
correlating	NN	O	O
with	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
activate	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
NF-AT	NN	O	B-protein
induction	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
were	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
inositol	NN	O	O
phosphate	NN	O	O
release	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
gross	NN	O	O
levels	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
induced	NN	O	O
late	NN	O	O
following	NN	O	O
the	NN	O	O
response	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
induction	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	O
expression	NN	O	O
occurred	NN	O	O
even	NN	O	O
with	NN	O	O
the	NN	O	O
smaller	NN	O	O
second	NN	O	O
messenger	NN	O	O
response	NN	O	O
generated	NN	O	O
by	NN	O	O
OKT3	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

HLA-DR-	NN	O	O
,	NN	O	O
CD33+	NN	O	O
,	NN	O	O
CD56+	NN	O	O
,	NN	O	O
CD16-	NN	O	O
myeloid/natural	NN	O	O
killer	NN	O	O
cell	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
:	NN	O	O
a	NN	O	O
previously	NN	O	O
unrecognized	NN	O	O
form	NN	O	O
of	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
potentially	NN	O	O
misdiagnosed	NN	O	O
as	NN	O	O
French-American-British	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia-M3	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
and	NN	O	O
characterized	NN	O	O
a	NN	O	O
previously	NN	O	O
unrecognized	NN	O	O
form	NN	O	O
of	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
that	NN	O	O
shares	NN	O	O
features	NN	O	O
of	NN	O	O
both	NN	O	O
myeloid	NN	O	B-cell_type
and	NN	O	I-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

From	NN	O	O
a	NN	O	O
consecutive	NN	O	O
series	NN	O	O
of	NN	O	O
350	NN	O	O
cases	NN	O	O
of	NN	O	O
adult	NN	O	O
de	NN	O	O
novo	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
(	NN	O	O
AML	NN	O	O
)	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
20	NN	O	O
cases	NN	O	O
(	NN	O	O
6	NN	O	O
%	NN	O	O
)	NN	O	O
with	NN	O	O
a	NN	O	O
unique	NN	O	O
immunophenotype	NN	O	O
:	NN	O	O
CD33+	NN	O	O
,	NN	O	O
CD56+	NN	O	O
,	NN	O	O
CD11a+	NN	O	O
,	NN	O	O
CD13lo	NN	O	O
,	NN	O	O
CD15lo	NN	O	O
,	NN	O	O
CD34+/-	NN	O	O
,	NN	O	O
HLA-DR	NN	O	B-protein
-	NN	O	O
,	NN	O	O
CD16	NN	O	B-protein
-	NN	O	O
.	NN	O	O

Multicolor	NN	O	O
flow	NN	O	O
cytometric	NN	O	O
assays	NN	O	O
confirmed	NN	O	O
the	NN	O	O
coexpression	NN	O	O
of	NN	O	O
myeloid	NN	O	B-protein
(	NN	O	I-protein
CD33	NN	O	I-protein
,	NN	O	I-protein
CD13	NN	O	I-protein
,	NN	O	I-protein
CD15	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
NK	NN	O	I-protein
cell-associated	NN	O	I-protein
(	NN	O	I-protein
CD56	NN	O	I-protein
)	NN	O	I-protein
antigens	NN	O	I-protein
in	NN	O	O
each	NN	O	O
case	NN	O	O
,	NN	O	O
whereas	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
assays	NN	O	O
confirmed	NN	O	O
the	NN	O	O
identity	NN	O	O
of	NN	O	O
CD56	NN	O	B-protein
(	NN	O	O
neural	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
)	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
blasts	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
two	NN	O	O
cases	NN	O	O
expressed	NN	O	O
CD4	NN	O	B-protein
,	NN	O	O
no	NN	O	O
case	NN	O	O
expressed	NN	O	O
CD2	NN	O	B-protein
,	NN	O	O
CD3	NN	O	B-protein
,	NN	O	O
or	NN	O	O
CD8	NN	O	B-protein
and	NN	O	O
no	NN	O	O
case	NN	O	O
showed	NN	O	O
clonal	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
genes	NN	O	O
encoding	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	I-protein
gamma	NN	O	I-protein
,	NN	O	I-protein
delta	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Leukemic	NN	O	O
blasts	NN	O	O
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
cases	NN	O	O
shared	NN	O	O
unique	NN	O	O
morphologic	NN	O	O
features	NN	O	O
(	NN	O	O
deeply	NN	O	O
invaginated	NN	O	O
nuclear	NN	O	O
membranes	NN	O	O
,	NN	O	O
scant	NN	O	O
cytoplasm	NN	O	O
with	NN	O	O
fine	NN	O	O
azurophilic	NN	O	O
granularity	NN	O	O
,	NN	O	O
and	NN	O	O
finely	NN	O	O
granular	NN	O	O
Sudan	NN	O	O
black	NN	O	O
B	NN	O	O
and	NN	O	O
myeloperoxidase	NN	O	B-protein
cytochemical	NN	O	O
reactivity	NN	O	O
)	NN	O	O
that	NN	O	O
were	NN	O	O
remarkably	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
;	NN	O	O
particularly	NN	O	O
the	NN	O	O
microgranular	NN	O	B-protein
variant	NN	O	I-protein
(	NN	O	O
FAB	NN	O	B-protein
AML-M3v	NN	O	I-protein
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
all	NN	O	O
20	NN	O	O
cases	NN	O	O
lacked	NN	O	O
the	NN	O	O
t	NN	O	O
(	NN	O	O
15	NN	O	O
;	NN	O	O
17	NN	O	O
)	NN	O	O
and	NN	O	O
17	NN	O	O
cases	NN	O	O
tested	NN	O	O
lacked	NN	O	O
the	NN	O	O
promyelocytic/retinoic	NN	O	B-RNA
acid	NN	O	I-RNA
receptor	NN	O	I-RNA
alpha	NN	O	I-RNA
(	NN	O	I-RNA
RAR	NN	O	I-RNA
alpha	NN	O	I-RNA
)	NN	O	I-RNA
fusion	NN	O	I-RNA
transcript	NN	O	I-RNA
in	NN	O	O
RT-PCR	NN	O	O
assays	NN	O	O
;	NN	O	O
12	NN	O	O
cases	NN	O	O
had	NN	O	O
46	NN	O	O
,	NN	O	O
XX	NN	O	O
or	NN	O	O
46	NN	O	O
,	NN	O	O
XY	NN	O	O
karyotypes	NN	O	O
,	NN	O	O
whereas	NN	O	O
2	NN	O	O
cases	NN	O	O
had	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
chromosome	NN	O	B-DNA
17q	NN	O	I-DNA
:	NN	O	O
1	NN	O	O
with	NN	O	O
del	NN	O	B-DNA
(	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q25	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
other	NN	O	O
with	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
11	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q23	NN	O	I-DNA
;	NN	O	I-DNA
q21	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
promyelocytic	NN	O	B-RNA
leukemia	NN	O	I-RNA
zinc	NN	O	I-RNA
finger/RAR	NN	O	I-RNA
alpha	NN	O	I-RNA
fusion	NN	O	I-RNA
transcript	NN	O	I-RNA
.	NN	O	O

All	NN	O	O
cases	NN	O	O
tested	NN	O	O
(	NN	O	O
6/20	NN	O	O
)	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
case	NN	O	O
with	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
11	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
failed	NN	O	O
to	NN	O	O
differentiate	NN	O	O
in	NN	O	O
vitro	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
cases	NN	O	O
may	NN	O	O
account	NN	O	O
for	NN	O	O
some	NN	O	O
APLs	NN	O	O
that	NN	O	O
have	NN	O	O
not	NN	O	O
shown	NN	O	O
a	NN	O	O
clinical	NN	O	O
response	NN	O	O
to	NN	O	O
ATRA	NN	O	O
.	NN	O	O

Four	NN	O	O
of	NN	O	O
6	NN	O	O
cases	NN	O	O
tested	NN	O	O
showed	NN	O	O
functional	NN	O	O
NK	NN	O	O
cell-mediated	NN	O	O
cytotoxicity	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
relationship	NN	O	O
between	NN	O	O
these	NN	O	O
unique	NN	O	O
CD33+	NN	O	O
,	NN	O	O
CD56+	NN	O	O
,	NN	O	O
CD16	NN	O	B-protein
-acute	NN	O	O
leukemias	NN	O	O
and	NN	O	O
normal	NN	O	B-cell_type
CD56+	NN	O	I-cell_type
,	NN	O	I-cell_type
CD16-	NN	O	I-cell_type
NK	NN	O	I-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
panning	NN	O	O
and	NN	O	O
multiparameter	NN	O	O
flow	NN	O	O
cytometric	NN	O	O
sorting	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
normal	NN	O	B-cell_type
CD56+	NN	O	I-cell_type
,	NN	O	I-cell_type
CD33+	NN	O	I-cell_type
,	NN	O	I-cell_type
CD16-	NN	O	I-cell_type
counterpart	NN	O	I-cell_type
cell	NN	O	I-cell_type
at	NN	O	O
a	NN	O	O
frequency	NN	O	O
of	NN	O	O
1	NN	O	O
%	NN	O	O
to	NN	O	O
2	NN	O	O
%	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
healthy	NN	O	O
individuals	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
this	NN	O	O
form	NN	O	O
of	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
may	NN	O	O
arise	NN	O	O
from	NN	O	O
transformation	NN	O	O
of	NN	O	O
a	NN	O	O
precursor	NN	O	O
cell	NN	O	O
common	NN	O	O
to	NN	O	O
both	NN	O	O
the	NN	O	O
myeloid	NN	O	B-cell_type
and	NN	O	I-cell_type
NK	NN	O	I-cell_type
cell	NN	O	I-cell_type
lineages	NN	O	I-cell_type
;	NN	O	O
thus	NN	O	O
we	NN	O	O
propose	NN	O	O
the	NN	O	O
designation	NN	O	O
myeloid/NK	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Recognition	NN	O	O
of	NN	O	O
this	NN	O	O
new	NN	O	O
leukemic	NN	O	O
entity	NN	O	O
will	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
distinguishing	NN	O	O
these	NN	O	O
ATRA-nonresponsive	NN	O	O
cases	NN	O	O
from	NN	O	O
ATRA-responsive	NN	O	O
true	NN	O	O
APL	NN	O	O
.	NN	O	O

-DOCSTART-	O

IL-4	NN	O	B-protein
down-regulates	NN	O	O
IL-2	NN	O	B-protein
-	NN	O	O
,	NN	O	O
IL-3	NN	O	B-protein
-	NN	O	O
,	NN	O	O
and	NN	O	O
GM-CSF	NN	O	B-protein
-induced	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
monocytes	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
,	NN	O	O
a	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
T-helper	NN	O	B-cell_type
0	NN	O	I-cell_type
(	NN	O	I-cell_type
Th0	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	I-cell_type
2	NN	O	I-cell_type
(	NN	O	I-cell_type
Th2	NN	O	I-cell_type
)	NN	O	I-cell_type
subset	NN	O	I-cell_type
,	NN	O	O
was	NN	O	O
originally	NN	O	O
described	NN	O	O
as	NN	O	O
a	NN	O	O
B-cell	NN	O	B-protein
stimulatory	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
has	NN	O	O
subsequently	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
suppress	NN	O	O
IL-1	NN	O	O
alpha	NN	O	O
,	NN	O	O
IL-1	NN	O	O
beta	NN	O	O
,	NN	O	O
IL-6	NN	O	O
,	NN	O	O
IL-8	NN	O	O
,	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
stimulated	NN	O	O
with	NN	O	O
LPS	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
upregulate	NN	O	O
IL-1	NN	O	B-DNA
receptor	NN	O	I-DNA
antagonist	NN	O	I-DNA
(	NN	O	I-DNA
IL1-RA	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
monocytes	NN	O	B-cell_type
evoked	NN	O	O
by	NN	O	O
other	NN	O	O
T-helper	NN	O	B-protein
cell	NN	O	I-protein
cytokines	NN	O	I-protein
:	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

IL-4	NN	O	B-protein
down-regulated	NN	O	O
mRNA	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
stimulated	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

IL-4	NN	O	B-protein
also	NN	O	O
suppressed	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
IL-6	NN	O	B-protein
mRNA	NN	O	O
expression	NN	O	O
.	NN	O	O

Temporal	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
down-regulatory	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
IL-2-	NN	O	O
,	NN	O	O
IL-3-	NN	O	O
,	NN	O	O
or	NN	O	O
GM-CSF-induced	NN	O	O
proinflammatory	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
provided	NN	O	O
evidence	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
acts	NN	O	O
predominantly	NN	O	O
on	NN	O	O
the	NN	O	O
post-transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
supported	NN	O	O
by	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
the	NN	O	O
down-regulatory	NN	O	O
capacity	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
dependent	NN	O	O
on	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
did	NN	O	O
not	NN	O	O
exert	NN	O	O
significant	NN	O	O
influence	NN	O	O
on	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-1-RA	NN	O	B-protein
or	NN	O	O
various	NN	O	O
CSFs	NN	O	B-protein
by	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
proto-oncogene	NN	O	B-DNA
and	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	B-cell_line
after	NN	O	O
stimulation	NN	O	O
with	NN	O	O
mycoplasma-derived	NN	O	O
material	NN	O	O
MDHM	NN	O	O
.	NN	O	O

Mycoplasma	NN	O	O
fermentans-derived	NN	O	O
high-molecular-weight	NN	O	O
material	NN	O	O
(	NN	O	O
MDHM	NN	O	O
)	NN	O	O
was	NN	O	O
originally	NN	O	O
described	NN	O	O
to	NN	O	O
induce	NN	O	O
differentiation	NN	O	O
of	NN	O	O
murine	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
to	NN	O	O
cytolytic	NN	O	B-cell_type
effector	NN	O	I-cell_type
T-cells	NN	O	I-cell_type
by	NN	O	O
stimulating	NN	O	O
IL-6	NN	O	B-protein
release	NN	O	O
from	NN	O	O
adherent	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
human	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
monocytes	NN	O	B-cell_type
(	NN	O	O
PBMo	NN	O	B-cell_type
)	NN	O	O
also	NN	O	O
respond	NN	O	O
to	NN	O	O
MDHM	NN	O	O
with	NN	O	O
increases	NN	O	O
in	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
both	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
level	NN	O	O
.	NN	O	O

The	NN	O	O
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-1	NN	O	B-RNA
beta	NN	O	I-RNA
and	NN	O	I-RNA
TNF	NN	O	I-RNA
alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
monocytic	NN	O	O
THP-1	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
increased	NN	O	O
as	NN	O	O
quickly	NN	O	O
as	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
PBMo	NN	O	B-cell_type
,	NN	O	O
THP-1	NN	O	B-cell_line
and	NN	O	O
14	NN	O	O
other	NN	O	O
monocytic/myeloid	NN	O	B-cell_line
leukemia-derived	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
did	NN	O	O
not	NN	O	O
secrete	NN	O	O
measurable	NN	O	O
amounts	NN	O	O
of	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
upon	NN	O	O
treatment	NN	O	O
with	NN	O	O
MDHM	NN	O	O
.	NN	O	O

IL-1	NN	O	B-DNA
beta	NN	O	I-DNA
and	NN	O	I-DNA
IL-6	NN	O	I-DNA
genes	NN	O	I-DNA
contain	NN	O	O
AP-1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
as	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
protein	NN	O	O
being	NN	O	O
composed	NN	O	O
of	NN	O	O
c-jun	NN	O	B-protein
and	NN	O	I-protein
c-fos	NN	O	I-protein
gene	NN	O	I-protein
products	NN	O	I-protein
.	NN	O	O

In	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
increased	NN	O	O
after	NN	O	O
incubation	NN	O	O
with	NN	O	O
MDHM	NN	O	O
while	NN	O	O
positive	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
remained	NN	O	O
unaffected	NN	O	O
.	NN	O	O

Although	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
AP-1	NN	O	B-protein
regulated	NN	O	O
cytokine	NN	O	B-protein
mRNA	NN	O	O
expression	NN	O	O
,	NN	O	O
results	NN	O	O
from	NN	O	O
PBMo	NN	O	B-cell_type
are	NN	O	O
not	NN	O	O
in	NN	O	O
accordance	NN	O	O
with	NN	O	O
this	NN	O	O
notion	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
primary	NN	O	B-cell_type
cells	NN	O	I-cell_type
MDHM-induced	NN	O	O
elevation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
was	NN	O	O
preceded	NN	O	O
by	NN	O	O
a	NN	O	O
downregulation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
while	NN	O	O
positive	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
was	NN	O	O
not	NN	O	O
modulated	NN	O	O
.	NN	O	O

c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
,	NN	O	O
constitutively	NN	O	O
high	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
was	NN	O	O
induced	NN	O	O
in	NN	O	O
MDHM-stimulated	NN	O	O
PBMo	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
MDHM-stimulated	NN	O	O
induction	NN	O	O
of	NN	O	O
cytokine	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
different	NN	O	O
proto-oncogene	NN	O	B-DNA
responses	NN	O	O
in	NN	O	O
PBMo	NN	O	B-cell_type
and	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
differences	NN	O	O
may	NN	O	O
represent	NN	O	O
different	NN	O	O
regulatory	NN	O	O
pathways	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
cell	NN	O	O
systems	NN	O	O
.	NN	O	O

Alternatively	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
neither	NN	O	O
AP-1	NN	O	B-protein
nor	NN	O	O
the	NN	O	O
c-myc	NN	O	B-protein
protein	NN	O	I-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
MDHM-induced	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-1	NN	O	O
beta	NN	O	O
,	NN	O	O
IL-6	NN	O	O
or	NN	O	O
TNF	NN	O	O
alpha	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
present	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
clearly	NN	O	O
that	NN	O	O
mycoplasma	NN	O	B-protein
products	NN	O	I-protein
can	NN	O	O
have	NN	O	O
a	NN	O	O
profound	NN	O	O
impact	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
status	NN	O	O
of	NN	O	O
eukaryotic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Novel	NN	O	B-protein
membrane	NN	O	I-protein
receptors	NN	O	I-protein
for	NN	O	O
aldosterone	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
a	NN	O	O
50	NN	O	B-protein
kDa	NN	O	I-protein
protein	NN	O	I-protein
on	NN	O	O
SDS-PAGE	NN	O	O
.	NN	O	O

Fast	NN	O	O
in	NN	O	O
vitro	NN	O	O
effects	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
on	NN	O	O
the	NN	O	O
Na+/H	NN	O	B-protein
(	NN	O	I-protein
+	NN	O	I-protein
)	NN	O	I-protein
-exchanger	NN	O	I-protein
,	NN	O	O
inositoltrisphosphate	NN	O	O
generation	NN	O	O
and	NN	O	O
corresponding	NN	O	O
specific	NN	O	O
binding	NN	O	O
to	NN	O	O
plasma	NN	O	O
membranes	NN	O	O
at	NN	O	O
Kd-values	NN	O	O
of	NN	O	O
approximately	NN	O	O
0.1	NN	O	O
nM	NN	O	O
have	NN	O	O
been	NN	O	O
found	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
and	NN	O	O
vascular	NN	O	B-cell_type
smooth	NN	O	I-cell_type
muscle	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
novel	NN	O	O
aldosterone	NN	O	O
membrane	NN	O	O
receptor	NN	O	O
was	NN	O	O
analyzed	NN	O	O
on	NN	O	O
SDS-PAGE	NN	O	O
after	NN	O	O
labeling	NN	O	O
of	NN	O	O
microsomal	NN	O	O
membranes	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
with	NN	O	O
a	NN	O	O
[	NN	O	O
125I	NN	O	O
]	NN	O	O
-aldosterone-derivative	NN	O	O
by	NN	O	O
use	NN	O	O
of	NN	O	O
BASED	NN	O	O
as	NN	O	O
a	NN	O	O
photoactivatable	NN	O	O
crosslinker	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
1	NN	O	O
nM	NN	O	O
[	NN	O	O
125I	NN	O	O
]	NN	O	O
-aldosterone	NN	O	O
was	NN	O	O
found	NN	O	O
at	NN	O	O
a	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
approximately	NN	O	O
50	NN	O	O
kDa	NN	O	O
which	NN	O	O
was	NN	O	O
absent	NN	O	O
with	NN	O	O
1	NN	O	O
microM	NN	O	O
cold	NN	O	O
aldosterone	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
cortisol	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
media	NN	O	O
.	NN	O	O

This	NN	O	O
aldosterone-selectivity	NN	O	O
is	NN	O	O
typical	NN	O	O
and	NN	O	O
discriminatory	NN	O	O
for	NN	O	O
the	NN	O	O
new	NN	O	O
aldosterone	NN	O	B-protein
membrane	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Solubilization	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
protein	NN	O	I-protein
from	NN	O	O
membranes	NN	O	O
by	NN	O	O
high	NN	O	O
salt	NN	O	O
concentrations	NN	O	O
(	NN	O	O
1	NN	O	O
M	NN	O	O
NaCl	NN	O	O
,	NN	O	O
1	NN	O	O
mM	NN	O	O
EDTA	NN	O	O
)	NN	O	O
was	NN	O	O
not	NN	O	O
achieved	NN	O	O
.	NN	O	O

It	NN	O	O
,	NN	O	O
thus	NN	O	O
,	NN	O	O
appears	NN	O	O
as	NN	O	O
an	NN	O	O
integral	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Dithiothreitol	NN	O	O
,	NN	O	O
a	NN	O	O
sulfhydryl	NN	O	O
agent	NN	O	O
,	NN	O	O
does	NN	O	O
not	NN	O	O
reduce	NN	O	O
specific	NN	O	O
aldosterone	NN	O	O
binding	NN	O	O
indicating	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
SH-groups	NN	O	B-protein
in	NN	O	O
the	NN	O	O
binding	NN	O	B-protein
domain	NN	O	I-protein
or	NN	O	O
sensitive	NN	O	O
structures	NN	O	O
of	NN	O	O
the	NN	O	O
receptors	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
are	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
novel	NN	O	B-protein
membrane	NN	O	I-protein
receptor	NN	O	I-protein
for	NN	O	O
aldosterone	NN	O	O
with	NN	O	O
regard	NN	O	O
to	NN	O	O
molecular	NN	O	O
weight	NN	O	O
and	NN	O	O
basic	NN	O	O
properties	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
and	NN	O	O
other	NN	O	O
related	NN	O	O
results	NN	O	O
are	NN	O	O
reviewed	NN	O	O
here	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
transcriptional	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
nuclease-hypersensitive	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
pol	NN	O	B-DNA
gene	NN	O	I-DNA
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
chromatin	NN	O	O
organization	NN	O	O
of	NN	O	O
the	NN	O	O
integrated	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	I-DNA
HIV-1	NN	O	I-DNA
)	NN	O	I-DNA
genome	NN	O	I-DNA
has	NN	O	O
previously	NN	O	O
revealed	NN	O	O
a	NN	O	O
major	NN	O	B-DNA
constitutive	NN	O	I-DNA
DNase	NN	O	I-DNA
I-hypersensitive	NN	O	I-DNA
site	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
the	NN	O	O
pol	NN	O	B-DNA
gene	NN	O	I-DNA
(	NN	O	O
E.	NN	O	O
Verdin	NN	O	O
,	NN	O	O
J.	NN	O	O
Virol.	NN	O	O
65	NN	O	O
:	NN	O	O
6790-6799	NN	O	O
,	NN	O	O
1991	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
report	NN	O	O
,	NN	O	O
high-resolution	NN	O	O
mapping	NN	O	O
of	NN	O	O
this	NN	O	O
site	NN	O	O
with	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	O
micrococcal	NN	O	B-protein
nuclease	NN	O	I-protein
identified	NN	O	O
a	NN	O	O
nucleosome-free	NN	O	B-DNA
region	NN	O	I-DNA
centered	NN	O	O
around	NN	O	O
nucleotides	NN	O	B-DNA
(	NN	O	I-DNA
nt	NN	O	I-DNA
)	NN	O	I-DNA
4490	NN	O	I-DNA
to	NN	O	I-DNA
4766	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
500-bp	NN	O	B-DNA
fragment	NN	O	I-DNA
encompassing	NN	O	O
this	NN	O	O
hypersensitive	NN	O	B-DNA
site	NN	O	I-DNA
(	NN	O	O
nt	NN	O	B-DNA
4481	NN	O	I-DNA
to	NN	O	I-DNA
4982	NN	O	I-DNA
)	NN	O	O
exhibited	NN	O	O
transcription-enhancing	NN	O	O
activity	NN	O	O
(	NN	O	O
two-	NN	O	O
to	NN	O	O
threefold	NN	O	O
)	NN	O	O
when	NN	O	O
it	NN	O	O
was	NN	O	O
cloned	NN	O	O
in	NN	O	O
its	NN	O	O
natural	NN	O	O
position	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
promoter	NN	O	I-DNA
after	NN	O	O
transient	NN	O	O
transfection	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
and	NN	O	O
CEM	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Using	NN	O	O
in	NN	O	O
vitro	NN	O	O
footprinting	NN	O	O
and	NN	O	O
gel	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
four	NN	O	O
distinct	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
within	NN	O	O
this	NN	O	O
positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Site	NN	O	B-DNA
B	NN	O	I-DNA
(	NN	O	O
nt	NN	O	B-DNA
4519	NN	O	I-DNA
to	NN	O	I-DNA
4545	NN	O	I-DNA
)	NN	O	O
specifically	NN	O	O
bound	NN	O	O
four	NN	O	O
distinct	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
complexes	NN	O	I-protein
:	NN	O	O
a	NN	O	O
ubiquitous	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
a	NN	O	O
T-cell-specific	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
monocyte/macrophage-	NN	O	B-protein
and	NN	O	I-protein
B-cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
PU.1/Spi-1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
most	NN	O	O
HIV-1	NN	O	O
isolates	NN	O	O
in	NN	O	O
which	NN	O	O
this	NN	O	O
PU	NN	O	B-DNA
box	NN	O	I-DNA
was	NN	O	O
not	NN	O	O
conserved	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
replaced	NN	O	O
by	NN	O	O
a	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
related	NN	O	O
factor	NN	O	O
Ets1	NN	O	B-protein
.	NN	O	O

Factors	NN	O	O
binding	NN	O	O
to	NN	O	O
site	NN	O	B-DNA
C	NN	O	I-DNA
(	NN	O	O
nt	NN	O	B-DNA
4681	NN	O	I-DNA
to	NN	O	I-DNA
4701	NN	O	I-DNA
)	NN	O	O
had	NN	O	O
a	NN	O	O
DNA-binding	NN	O	O
specificity	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
factors	NN	O	O
binding	NN	O	O
to	NN	O	O
site	NN	O	B-DNA
B	NN	O	I-DNA
,	NN	O	O
except	NN	O	O
for	NN	O	O
PU.1/Spi-1	NN	O	B-protein
.	NN	O	O

A	NN	O	O
GC	NN	O	B-DNA
box	NN	O	I-DNA
containing	NN	O	O
a	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
Sp1	NN	O	B-protein
was	NN	O	O
identified	NN	O	O
(	NN	O	O
nt	NN	O	B-DNA
4623	NN	O	I-DNA
to	NN	O	I-DNA
4631	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

Site	NN	O	B-DNA
D	NN	O	I-DNA
(	NN	O	O
nt	NN	O	B-DNA
4816	NN	O	I-DNA
to	NN	O	I-DNA
4851	NN	O	I-DNA
)	NN	O	O
specifically	NN	O	O
bound	NN	O	O
a	NN	O	O
ubiquitously	NN	O	B-protein
expressed	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
identify	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
a	NN	O	O
nuclease-hypersensitive	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
pol	NN	O	B-DNA
gene	NN	O	I-DNA
of	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
its	NN	O	O
activity	NN	O	O
may	NN	O	O
be	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
complex	NN	O	O
interplay	NN	O	O
of	NN	O	O
cis-	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
v-src	NN	O	B-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
correlates	NN	O	O
with	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
rapidly	NN	O	O
inducible	NN	O	O
transcriptional	NN	O	B-DNA
activator	NN	O	I-DNA
that	NN	O	O
responds	NN	O	O
to	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
signals	NN	O	O
and	NN	O	O
influences	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Protein	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
transmit	NN	O	O
signals	NN	O	O
from	NN	O	O
cytokine	NN	O	B-protein
and	NN	O	I-protein
immune	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Very	NN	O	O
little	NN	O	O
information	NN	O	O
exists	NN	O	O
linking	NN	O	O
these	NN	O	O
two	NN	O	O
important	NN	O	O
classes	NN	O	O
of	NN	O	O
signaling	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

We	NN	O	O
now	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
v-src	NN	O	B-DNA
expression	NN	O	O
correlates	NN	O	O
with	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	I-protein
complex	NN	O	I-protein
similar	NN	O	O
to	NN	O	O
that	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
,	NN	O	O
as	NN	O	O
detected	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
using	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
complex	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
inhibitor	NN	O	O
,	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
.	NN	O	O

The	NN	O	O
v-src-induced	NN	O	B-protein
complex	NN	O	I-protein
comprised	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
components	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
supershift	NN	O	O
and	NN	O	O
immunoblot	NN	O	O
analysis	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
functional	NN	O	O
correlate	NN	O	O
of	NN	O	O
this	NN	O	O
finding	NN	O	O
,	NN	O	O
transient	NN	O	O
co-transfection	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
different	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
demonstrated	NN	O	O
that	NN	O	O
v-src	NN	O	B-DNA
activated	NN	O	O
this	NN	O	O
promoter	NN	O	B-protein
in	NN	O	O
a	NN	O	O
kappa	NN	O	O
B-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
by	NN	O	O
v-src	NN	O	B-DNA
was	NN	O	O
more	NN	O	O
sensitive	NN	O	O
to	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
proximal	NN	O	O
,	NN	O	O
rather	NN	O	O
than	NN	O	O
the	NN	O	O
distal	NN	O	O
,	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
implications	NN	O	O
for	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
signaling	NN	O	O
and	NN	O	O
HIV-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
are	NN	O	O
considered	NN	O	O
.	NN	O	O

-DOCSTART-	O

trans-activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
a	NN	O	O
cDNA	NN	O	O
and	NN	O	O
its	NN	O	O
genomic	NN	O	O
clones	NN	O	O
of	NN	O	O
human	NN	O	O
herpesvirus-6	NN	O	O
.	NN	O	O

Human	NN	O	O
herpesvirus	NN	O	O
6	NN	O	O
(	NN	O	O
HHV-6	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
lymphotropic	NN	O	O
herpesvirus	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
it	NN	O	O
can	NN	O	O
productively	NN	O	O
infect	NN	O	O
human	NN	O	O
CD4+	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

Co-infection	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
HHV-6	NN	O	O
can	NN	O	O
lead	NN	O	O
to	NN	O	O
both	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
acceleration	NN	O	O
of	NN	O	O
the	NN	O	O
cytopathic	NN	O	O
effects	NN	O	O
.	NN	O	O

An	NN	O	O
HHV-6	NN	O	B-DNA
(	NN	O	I-DNA
GS	NN	O	I-DNA
)	NN	O	I-DNA
cDNA	NN	O	I-DNA
clone	NN	O	I-DNA
,	NN	O	O
pCD41	NN	O	B-DNA
,	NN	O	O
encoding	NN	O	O
for	NN	O	O
a	NN	O	O
41-kDa	NN	O	B-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
identified	NN	O	O
and	NN	O	O
characterized	NN	O	O
previously	NN	O	O
(	NN	O	O
Chang	NN	O	O
and	NN	O	O
Balachandran	NN	O	O
,	NN	O	O
J.	NN	O	O
Virol.	NN	O	O
65	NN	O	O
,	NN	O	O
2884-2894	NN	O	O
and	NN	O	O
7085	NN	O	O
,	NN	O	O
1991	NN	O	O
)	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analyses	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
protein	NN	O	O
has	NN	O	O
significant	NN	O	O
homology	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
cytomegalovirus	NN	O	I-DNA
UL44	NN	O	I-DNA
gene	NN	O	I-DNA
coding	NN	O	O
for	NN	O	O
the	NN	O	O
ICP36	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
early-late-class	NN	O	B-protein
phosphoprotein	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
this	NN	O	O
cDNA	NN	O	B-DNA
as	NN	O	O
the	NN	O	O
probe	NN	O	O
,	NN	O	O
a	NN	O	O
3.8-kb	NN	O	B-DNA
EcoRI	NN	O	I-DNA
genomic	NN	O	I-DNA
fragment	NN	O	I-DNA
encoding	NN	O	O
the	NN	O	O
HHV-6	NN	O	B-protein
(	NN	O	I-protein
GS	NN	O	I-protein
)	NN	O	I-protein
P41	NN	O	I-protein
was	NN	O	O
cloned	NN	O	O
and	NN	O	O
designated	NN	O	O
as	NN	O	O
pGD41	NN	O	B-DNA
.	NN	O	O

When	NN	O	O
cotransfected	NN	O	O
with	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
CAT	NN	O	I-DNA
into	NN	O	O
CV-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
both	NN	O	O
the	NN	O	O
pCD41	NN	O	B-DNA
and	NN	O	O
pGD41	NN	O	B-DNA
clones	NN	O	O
trans-activated	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

Sequence	NN	O	O
analyses	NN	O	O
of	NN	O	O
pCD41	NN	O	B-DNA
indicate	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
two	NN	O	O
potential	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frames	NN	O	I-DNA
(	NN	O	O
ORFs	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
A	NN	O	B-DNA
and	NN	O	O
B	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
are	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
ORFs	NN	O	B-DNA
found	NN	O	O
in	NN	O	O
the	NN	O	O
genomic	NN	O	O
clone	NN	O	O
pGD41	NN	O	B-DNA
.	NN	O	O

Deletion	NN	O	O
constructs	NN	O	O
of	NN	O	O
the	NN	O	O
pCD41	NN	O	B-DNA
clone	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
ORF-A	NN	O	B-protein
was	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

Deletion	NN	O	O
analyses	NN	O	O
of	NN	O	O
the	NN	O	O
pCD41	NN	O	B-DNA
ORF-A	NN	O	B-protein
and	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
promoter	NN	O	B-DNA
constructs	NN	O	I-DNA
further	NN	O	O
mapped	NN	O	O
an	NN	O	O
internal	NN	O	O
functional	NN	O	O
promoter	NN	O	O
within	NN	O	O
the	NN	O	O
pCD41	NN	O	B-DNA
sequence	NN	O	O
that	NN	O	O
can	NN	O	O
direct	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
the	NN	O	O
trans-activating	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

By	NN	O	O
using	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
deletion	NN	O	I-DNA
mutants	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	B-DNA
sites	NN	O	I-DNA
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
critical	NN	O	O
for	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
pCD41	NN	O	B-DNA
trans-activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD14	NN	O	B-protein
-mediated	NN	O	O
translocation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
lipopolysaccharide	NN	O	O
does	NN	O	O
not	NN	O	O
require	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

During	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
serious	NN	O	O
bacterial	NN	O	O
infections	NN	O	O
,	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
macrophage	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
mediators	NN	O	O
and	NN	O	O
systemic	NN	O	O
symptoms	NN	O	O
including	NN	O	O
hemodynamic	NN	O	O
instability	NN	O	O
and	NN	O	O
shock	NN	O	O
.	NN	O	O

CD14	NN	O	B-protein
,	NN	O	O
a	NN	O	O
glycosylphosphatidylinositol-linked	NN	O	B-protein
antigen	NN	O	I-protein
,	NN	O	O
functions	NN	O	O
as	NN	O	O
an	NN	O	O
LPS	NN	O	B-protein
signaling	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

A	NN	O	O
critical	NN	O	O
issue	NN	O	O
concerns	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
CD14	NN	O	B-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
no	NN	O	O
transmembrane	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
transduces	NN	O	O
its	NN	O	O
signal	NN	O	O
following	NN	O	O
LPS	NN	O	O
binding	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
investigators	NN	O	O
have	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
CD14	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
is	NN	O	O
effected	NN	O	O
through	NN	O	O
a	NN	O	O
receptor-associated	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
TK	NN	O	O
)	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
multicomponent	NN	O	O
receptor	NN	O	O
model	NN	O	O
of	NN	O	O
LPS	NN	O	O
signaling	NN	O	O
.	NN	O	O

Wild-type	NN	O	O
Chinese	NN	O	B-cell_line
hamster	NN	O	I-cell_line
ovary	NN	O	I-cell_line
(	NN	O	I-cell_line
CHO	NN	O	I-cell_line
)	NN	O	I-cell_line
-K1	NN	O	I-cell_line
cells	NN	O	I-cell_line
can	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
endotoxin	NN	O	O
to	NN	O	O
release	NN	O	O
arachidonate	NN	O	O
following	NN	O	O
transfection	NN	O	O
with	NN	O	O
human	NN	O	O
CD14	NN	O	B-protein
(	NN	O	O
CHO/CD14	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
cytosolic	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
LPS-inducible	NN	O	O
responses	NN	O	O
.	NN	O	O

We	NN	O	O
sought	NN	O	O
to	NN	O	O
determine	NN	O	O
if	NN	O	O
this	NN	O	O
pathway	NN	O	O
were	NN	O	O
present	NN	O	O
in	NN	O	O
CHO/CD14	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
relationship	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
to	NN	O	O
the	NN	O	O
CD14	NN	O	B-protein
receptor	NN	O	O
system	NN	O	O
.	NN	O	O

LPS-stimulated	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
CHO/CD14	NN	O	B-cell_line
cells	NN	O	I-cell_line
resembled	NN	O	O
the	NN	O	O
same	NN	O	O
response	NN	O	O
in	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
macrophage-like	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
RAW	NN	O	B-cell_line
264.7	NN	O	I-cell_line
.	NN	O	O

Protein	NN	O	O
synthesis	NN	O	O
inhibitors	NN	O	O
and	NN	O	O
corticosteroids	NN	O	O
,	NN	O	O
which	NN	O	O
suppress	NN	O	O
arachidonate	NN	O	O
release	NN	O	O
and	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
CHO/	NN	O	O
CD14	NN	O	B-protein
or	NN	O	O
RAW	NN	O	B-cell_line
264.7	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
is	NN	O	O
an	NN	O	O
early	NN	O	O
event	NN	O	O
.	NN	O	O

Although	NN	O	O
TK	NN	O	O
activity	NN	O	O
was	NN	O	O
consistently	NN	O	O
observed	NN	O	O
by	NN	O	O
immunoblotting	NN	O	O
extracts	NN	O	O
from	NN	O	O
activated	NN	O	O
RAW	NN	O	B-cell_line
264.7	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
LPS-induced	NN	O	O
phosphotyrosine	NN	O	O
residues	NN	O	O
were	NN	O	O
not	NN	O	O
observed	NN	O	O
from	NN	O	O
similarly	NN	O	O
treated	NN	O	O
CHO/CD14	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
TK	NN	O	O
inhibitors	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
and	NN	O	O
genistein	NN	O	O
failed	NN	O	O
to	NN	O	O
inhibit	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
CHO/CD14	NN	O	B-cell_line
or	NN	O	O
RAW	NN	O	B-cell_line
264.7	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
both	NN	O	O
of	NN	O	O
these	NN	O	O
agents	NN	O	O
inhibited	NN	O	O
LPS-induced	NN	O	O
TK	NN	O	O
activity	NN	O	O
in	NN	O	O
RAW	NN	O	B-cell_line
264.7	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
imply	NN	O	O
that	NN	O	O
TK	NN	O	O
activity	NN	O	O
is	NN	O	O
not	NN	O	O
obligatory	NN	O	O
for	NN	O	O
CD14	NN	O	B-protein
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
to	NN	O	O
occur	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signals	NN	O	O
transduced	NN	O	O
through	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
molecule	NN	O	I-protein
on	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
native	NN	O	B-protein
envelope	NN	O	I-protein
glycoproteins	NN	O	I-protein
of	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
gp160	NN	O	O
can	NN	O	O
induce	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
stimulatory	NN	O	O
effects	NN	O	O
of	NN	O	O
gp160	NN	O	O
are	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
molecule	NN	O	I-protein
,	NN	O	O
since	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
soluble	NN	O	O
CD4	NN	O	B-protein
abrogates	NN	O	O
its	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
gp160-induced	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
complex	NN	O	I-protein
consists	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
c-rel	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
stimulatory	NN	O	O
effect	NN	O	O
of	NN	O	O
gp160	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
is	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
independent	NN	O	O
,	NN	O	O
is	NN	O	O
dependent	NN	O	O
upon	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
and	NN	O	O
abrogated	NN	O	O
by	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
.	NN	O	O

The	NN	O	O
gp160-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
CD4	NN	O	B-cell_type
positive	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
biological	NN	O	O
effects	NN	O	O
,	NN	O	O
e.g.	NN	O	O
,	NN	O	O
enhanced	NN	O	O
HIV	NN	O	O
replication	NN	O	O
,	NN	O	O
hypergammaglobulinemia	NN	O	O
,	NN	O	O
increased	NN	O	O
cytokine	NN	O	B-protein
secretion	NN	O	O
,	NN	O	O
hypercellularity	NN	O	O
in	NN	O	O
bone	NN	O	O
marrow	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

No	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
on	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
during	NN	O	O
pregnancy	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
was	NN	O	O
analysed	NN	O	O
,	NN	O	O
using	NN	O	O
an	NN	O	O
enzyme	NN	O	O
linked	NN	O	O
immunosorbent	NN	O	O
assay	NN	O	O
(	NN	O	O
Abbott	NN	O	O
PgR-EIA	NN	O	O
monoclonal	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
evaluate	NN	O	O
its	NN	O	O
prognostic	NN	O	O
character	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
spontaneous	NN	O	O
abortion	NN	O	O
.	NN	O	O

Cytosols	NN	O	O
were	NN	O	O
prepared	NN	O	O
from	NN	O	O
lymphocytes	NN	O	O
of	NN	O	O
24	NN	O	O
healthy	NN	O	O
pregnant	NN	O	O
women	NN	O	O
(	NN	O	O
11	NN	O	O
first	NN	O	O
,	NN	O	O
10	NN	O	O
second	NN	O	O
and	NN	O	O
three	NN	O	O
third	NN	O	O
trimester	NN	O	O
)	NN	O	O
,	NN	O	O
seven	NN	O	O
healthy	NN	O	O
non-pregnant	NN	O	O
women	NN	O	O
,	NN	O	O
nine	NN	O	O
women	NN	O	O
with	NN	O	O
recurrent	NN	O	O
spontaneous	NN	O	O
abortion	NN	O	O
,	NN	O	O
and	NN	O	O
six	NN	O	O
healthy	NN	O	O
men	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
breast	NN	O	I-cell_line
carcinoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
ZR-75-1	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
which	NN	O	O
expresses	NN	O	O
the	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
was	NN	O	O
analysed	NN	O	O
throughout	NN	O	O
.	NN	O	O

The	NN	O	O
ZR-75-1	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
showed	NN	O	O
an	NN	O	O
expression	NN	O	O
of	NN	O	O
642	NN	O	O
fmol/mg	NN	O	O
whereas	NN	O	O
lymphocytes	NN	O	O
of	NN	O	O
pregnant	NN	O	O
women	NN	O	O
showed	NN	O	O
an	NN	O	O
expression	NN	O	O
<	NN	O	O
or	NN	O	O
=	NN	O	O
4	NN	O	O
fmol/mg	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
of	NN	O	O
non-pregnant	NN	O	O
women	NN	O	O
,	NN	O	O
women	NN	O	O
with	NN	O	O
threatened	NN	O	O
pre-term	NN	O	O
delivery	NN	O	O
,	NN	O	O
and	NN	O	O
men	NN	O	O
showed	NN	O	O
equivalent	NN	O	O
levels	NN	O	O
:	NN	O	O
3	NN	O	O
+/-	NN	O	O
1	NN	O	O
,	NN	O	O
3	NN	O	O
+/-	NN	O	O
2	NN	O	O
and	NN	O	O
5	NN	O	O
+/-	NN	O	O
4	NN	O	O
fmol/mg	NN	O	O
respectively	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
evidence	NN	O	O
of	NN	O	O
specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
pregnancy	NN	O	O
and	NN	O	O
exclude	NN	O	O
any	NN	O	O
prognostic	NN	O	O
character	NN	O	O
in	NN	O	O
spontaneous	NN	O	O
abortion	NN	O	O
.	NN	O	O

A	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
known	NN	O	O
effect	NN	O	O
of	NN	O	O
progesterone	NN	O	O
on	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
also	NN	O	O
excluded	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tolerance	NN	O	O
to	NN	O	O
lipopolysaccharide	NN	O	O
involves	NN	O	O
mobilization	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
with	NN	O	O
predominance	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
with	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
leads	NN	O	O
to	NN	O	O
rapid	NN	O	O
and	NN	O	O
transient	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
like	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

When	NN	O	O
such	NN	O	O
cells	NN	O	O
are	NN	O	O
precultured	NN	O	O
for	NN	O	O
2	NN	O	O
days	NN	O	O
with	NN	O	O
a	NN	O	O
low	NN	O	O
dose	NN	O	O
of	NN	O	O
LPS	NN	O	O
(	NN	O	O
20	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
followed	NN	O	O
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
a	NN	O	O
high	NN	O	O
dose	NN	O	O
of	NN	O	O
LPS	NN	O	O
(	NN	O	O
1	NN	O	O
microgram/ml	NN	O	O
)	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
minimal	NN	O	O
,	NN	O	O
i.e.	NN	O	O
the	NN	O	O
cells	NN	O	O
are	NN	O	O
tolerant	NN	O	O
.	NN	O	O

In	NN	O	O
nuclear	NN	O	O
run-on	NN	O	O
analysis	NN	O	O
,	NN	O	O
such	NN	O	O
tolerant	NN	O	B-cell_type
cells	NN	O	I-cell_type
show	NN	O	O
only	NN	O	O
a	NN	O	O
low	NN	O	O
degree	NN	O	O
of	NN	O	O
transcription	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
tolerance	NN	O	O
operates	NN	O	O
at	NN	O	O
or	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
level	NN	O	O
.	NN	O	O

The	NN	O	O
CD14	NN	O	B-protein
LPS	NN	O	O
receptor	NN	O	O
is	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
up-regulated	NN	O	O
(	NN	O	O
not	NN	O	O
down-regulated	NN	O	O
)	NN	O	O
in	NN	O	O
tolerant	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
LPS	NN	O	O
can	NN	O	O
,	NN	O	O
in	NN	O	O
fact	NN	O	O
,	NN	O	O
still	NN	O	O
lead	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
tolerant	NN	O	B-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
mobilization	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Resolution	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
gel	NN	O	O
shift	NN	O	O
analysis	NN	O	O
shows	NN	O	O
that	NN	O	O
the	NN	O	O
binding	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
mobilized	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_line
Mono	NN	O	I-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
consists	NN	O	O
mainly	NN	O	O
of	NN	O	O
p50-p65	NN	O	B-protein
heterodimers	NN	O	I-protein
,	NN	O	O
while	NN	O	O
in	NN	O	O
tolerant	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
homodimer	NN	O	I-protein
is	NN	O	O
predominant	NN	O	O
.	NN	O	O

This	NN	O	O
increase	NN	O	O
in	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	I-protein
coincides	NN	O	O
with	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
p105	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
suggestive	NN	O	O
of	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
.	NN	O	O

Reporter	NN	O	O
gene	NN	O	O
analysis	NN	O	O
reveals	NN	O	O
that	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
mobilized	NN	O	O
in	NN	O	O
tolerant	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
functionally	NN	O	O
inactive	NN	O	O
in	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-DNA
B-dependent	NN	O	I-DNA
luciferase	NN	O	I-DNA
constructs	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
or	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
5'-region	NN	O	I-DNA
show	NN	O	O
only	NN	O	O
minimal	NN	O	O
transactivation	NN	O	O
after	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
primary	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
when	NN	O	O
precultured	NN	O	O
with	NN	O	O
a	NN	O	O
low	NN	O	O
dose	NN	O	O
of	NN	O	O
LPS	NN	O	O
,	NN	O	O
also	NN	O	O
become	NN	O	O
tolerant	NN	O	O
and	NN	O	O
produce	NN	O	O
little	NN	O	O
TNF	NN	O	B-protein
after	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
tolerant	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
also	NN	O	O
up-regulate	NN	O	O
CD14	NN	O	B-protein
,	NN	O	O
and	NN	O	O
they	NN	O	O
mobilize	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
with	NN	O	O
a	NN	O	O
predominance	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	I-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
tolerance	NN	O	O
to	NN	O	O
LPS	NN	O	O
is	NN	O	O
determined	NN	O	O
by	NN	O	O
post-receptor	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
involve	NN	O	O
an	NN	O	O
altered	NN	O	O
composition	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Analysis	NN	O	O
of	NN	O	O
Oct2-isoform	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Oct2-isoform	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
splenic	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
lipopolysaccharide	NN	O	O
or	NN	O	O
lipopolysaccharide	NN	O	O
plus	NN	O	O
phorbol-di-butyrate	NN	O	O
was	NN	O	O
analysed	NN	O	O
by	NN	O	O
cDNA	NN	O	O
cloning	NN	O	O
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
of	NN	O	O
Oct2-positive	NN	O	B-cell_line
clones	NN	O	I-cell_line
was	NN	O	O
1/15	NN	O	O
,	NN	O	O
000	NN	O	O
in	NN	O	O
both	NN	O	O
libraries	NN	O	O
.	NN	O	O

Two	NN	O	O
new	NN	O	O
isoforms	NN	O	O
were	NN	O	O
found	NN	O	O
that	NN	O	O
generate	NN	O	O
novel	NN	O	O
amino-	NN	O	B-protein
or	NN	O	I-protein
carboxy-terminal	NN	O	I-protein
sequences	NN	O	I-protein
.	NN	O	O

An	NN	O	O
isoform	NN	O	O
lacking	NN	O	O
exon	NN	O	B-DNA
11	NN	O	I-DNA
destroyed	NN	O	O
the	NN	O	O
carboxy-terminal	NN	O	B-protein
leucin-zipper	NN	O	I-protein
region	NN	O	I-protein
and	NN	O	O
introduced	NN	O	O
a	NN	O	O
frame	NN	O	O
shift	NN	O	O
creating	NN	O	O
a	NN	O	O
novel	NN	O	O
,	NN	O	O
proline-rich	NN	O	B-protein
carboxy	NN	O	I-protein
terminus	NN	O	I-protein
.	NN	O	O

A	NN	O	O
new	NN	O	O
exon	NN	O	B-DNA
containing	NN	O	O
a	NN	O	O
highly	NN	O	O
basic	NN	O	O
region	NN	O	O
(	NN	O	O
4c	NN	O	O
)	NN	O	O
was	NN	O	O
characterized	NN	O	O
,	NN	O	O
between	NN	O	B-DNA
exons	NN	O	I-DNA
4	NN	O	I-DNA
and	NN	O	I-DNA
5	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
exon	NN	O	O
was	NN	O	O
inserted	NN	O	O
between	NN	O	B-DNA
glutamine-rich	NN	O	I-DNA
regions	NN	O	I-DNA
2	NN	O	I-DNA
and	NN	O	I-DNA
3	NN	O	I-DNA
,	NN	O	O
carboxy	NN	O	B-protein
terminal	NN	O	I-protein
of	NN	O	O
a	NN	O	O
tentative	NN	O	O
leucine-zipper	NN	O	B-protein
structure	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
a	NN	O	O
new	NN	O	O
combination	NN	O	O
isoform	NN	O	O
containing	NN	O	O
Oct2a	NN	O	B-protein
's	NN	O	I-protein
amino	NN	O	I-protein
terminal	NN	O	I-protein
insert	NN	O	I-protein
(	NN	O	O
exon	NN	O	B-DNA
7a	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
Oct2b	NN	O	B-protein
's	NN	O	I-protein
carboxy	NN	O	I-protein
terminal	NN	O	I-protein
insert	NN	O	I-protein
(	NN	O	O
exon	NN	O	B-DNA
13	NN	O	I-DNA
)	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
created	NN	O	O
a	NN	O	O
novel	NN	O	O
large	NN	O	O
isoform	NN	O	O
,	NN	O	O
Oct2ab	NN	O	B-protein
.	NN	O	O

More	NN	O	O
frequent	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
classical	NN	O	O
Oct2a	NN	O	B-protein
and	NN	O	O
Oct2b	NN	O	B-protein
isoforms	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
a	NN	O	O
preference	NN	O	O
for	NN	O	O
the	NN	O	O
Oct2ab	NN	O	B-protein
and	NN	O	O
Oct2ba	NN	O	B-protein
isoforms	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
lipopolysaccharide	NN	O	B-cell_line
plus	NN	O	I-cell_line
phorbol-di-butyrate-treated	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Positive	NN	O	O
regulators	NN	O	O
of	NN	O	O
the	NN	O	O
lineage-specific	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
in	NN	O	O
differentiating	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
major	NN	O	O
regulator	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
,	NN	O	I-cell_type
megakaryocyte	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
mast	NN	O	I-cell_type
cell	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

GATA-1	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
WGATAR	NN	O	B-DNA
consensus	NN	O	I-DNA
motifs	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
regulatory	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
virtually	NN	O	O
all	NN	O	O
erythroid	NN	O	B-DNA
cell-specific	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Analyses	NN	O	O
with	NN	O	O
cultured	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
cell-free	NN	O	O
systems	NN	O	O
have	NN	O	O
provided	NN	O	O
strong	NN	O	O
evidence	NN	O	O
that	NN	O	O
GATA-1	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
control	NN	O	O
of	NN	O	O
globin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
during	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Targeted	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
GATA-1	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
embryonic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
demonstrated	NN	O	O
its	NN	O	O
requirement	NN	O	O
in	NN	O	O
normal	NN	O	O
erythroid	NN	O	O
development	NN	O	O
.	NN	O	O

Efficient	NN	O	O
rescue	NN	O	O
of	NN	O	O
the	NN	O	O
defect	NN	O	O
requires	NN	O	O
an	NN	O	O
intact	NN	O	O
GATA	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
autoregulatory	NN	O	O
control	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
whether	NN	O	O
GATA-1	NN	O	B-protein
expression	NN	O	O
involves	NN	O	O
additional	NN	O	O
regulatory	NN	O	B-protein
factors	NN	O	I-protein
or	NN	O	O
is	NN	O	O
maintained	NN	O	O
entirely	NN	O	O
by	NN	O	O
an	NN	O	O
autoregulatory	NN	O	O
loop	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
used	NN	O	O
a	NN	O	O
transient	NN	O	O
heterokaryon	NN	O	O
system	NN	O	O
to	NN	O	O
test	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
erythroid	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
activate	NN	O	O
the	NN	O	O
GATA-1	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
nonerythroid	NN	O	O
nuclei	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
proerythroblasts	NN	O	B-cell_type
and	NN	O	O
mature	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
contain	NN	O	O
a	NN	O	O
diffusible	NN	O	O
activity	NN	O	O
(	NN	O	O
TAG	NN	O	O
)	NN	O	O
capable	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
that	NN	O	O
this	NN	O	O
activity	NN	O	O
decreases	NN	O	O
during	NN	O	O
the	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Nuclei	NN	O	O
from	NN	O	O
GATA-1	NN	O	B-cell_line
-mutant	NN	O	I-cell_line
embryonic	NN	O	I-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
can	NN	O	O
still	NN	O	O
be	NN	O	O
reprogrammed	NN	O	O
to	NN	O	O
express	NN	O	O
their	NN	O	O
globin	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
erythroid	NN	O	B-cell_type
heterokaryons	NN	O	I-cell_type
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
de	NN	O	O
novo	NN	O	O
induction	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
globin	NN	O	B-DNA
gene	NN	O	I-DNA
activation	NN	O	O
following	NN	O	O
cell	NN	O	O
fusion	NN	O	O
.	NN	O	O

-DOCSTART-	O

Role	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-protein
Nef	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
activation	NN	O	O
pathways	NN	O	O
in	NN	O	O
CD4	NN	O	B-protein
+	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
immunodeficiency	NN	O	I-protein
virus	NN	O	I-protein
(	NN	O	I-protein
HIV-1	NN	O	I-protein
)	NN	O	I-protein
Nef	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
pathways	NN	O	O
was	NN	O	O
investigated	NN	O	O
using	NN	O	O
a	NN	O	O
Jurkat	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
Nef	NN	O	O
expression	NN	O	O
vector	NN	O	O
.	NN	O	O

Secretion	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
,	NN	O	O
and	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
(	NN	O	I-protein
Fos/Jun	NN	O	I-protein
)	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
or	NN	O	O
immobilized	NN	O	O
antibodies	NN	O	O
to	NN	O	O
CD3	NN	O	B-protein
were	NN	O	O
monitored	NN	O	O
.	NN	O	O

These	NN	O	O
parameters	NN	O	O
were	NN	O	O
not	NN	O	O
modified	NN	O	O
by	NN	O	O
Nef	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
stimulation	NN	O	O
with	NN	O	O
the	NN	O	O
same	NN	O	O
stimuli	NN	O	O
resulted	NN	O	O
in	NN	O	O
partial	NN	O	O
inhibition	NN	O	O
of	NN	O	O
LTR	NN	O	B-DNA
activation	NN	O	O
in	NN	O	O
Nef+	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
was	NN	O	O
not	NN	O	O
mediated	NN	O	O
through	NN	O	O
Nef	NN	O	B-protein
phosphorylation	NN	O	O
on	NN	O	O
Thr-15	NN	O	O
or	NN	O	O
GTP-binding	NN	O	O
activity	NN	O	O
because	NN	O	O
mutations	NN	O	O
in	NN	O	O
critical	NN	O	O
sites	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
this	NN	O	O
inhibition	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
truncated	NN	O	O
LTRs	NN	O	B-DNA
confirmed	NN	O	O
that	NN	O	O
inhibition	NN	O	O
of	NN	O	O
LTR	NN	O	B-DNA
activation	NN	O	O
was	NN	O	O
not	NN	O	O
mediated	NN	O	O
through	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-binding	NN	O	O
activity	NN	O	O
but	NN	O	O
through	NN	O	O
the	NN	O	O
region	NN	O	O
containing	NN	O	O
the	NN	O	O
negative	NN	O	B-DNA
responding	NN	O	I-DNA
elements	NN	O	I-DNA
(	NN	O	O
NREs	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
Nef	NN	O	B-protein
downmodulates	NN	O	O
LTR	NN	O	B-DNA
activation	NN	O	O
without	NN	O	O
significantly	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
capacity	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
respond	NN	O	O
to	NN	O	O
immunological	NN	O	O
activations	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tat-binding	NN	O	B-protein
protein	NN	O	I-protein
7	NN	O	I-protein
is	NN	O	O
a	NN	O	O
subunit	NN	O	O
of	NN	O	O
the	NN	O	O
26S	NN	O	B-protein
protease	NN	O	I-protein
.	NN	O	O

Subunit	NN	O	B-protein
6	NN	O	I-protein
(	NN	O	O
S6	NN	O	B-protein
)	NN	O	O
,	NN	O	O
an	NN	O	O
integral	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
26S	NN	O	B-protein
protease	NN	O	I-protein
from	NN	O	O
human	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
,	NN	O	O
has	NN	O	O
been	NN	O	O
studied	NN	O	O
by	NN	O	O
SDS-PAGE	NN	O	O
,	NN	O	O
peptide	NN	O	O
mapping	NN	O	O
and	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
.	NN	O	O

S6	NN	O	B-protein
was	NN	O	O
cleaved	NN	O	O
with	NN	O	O
CNBr	NN	O	B-protein
and	NN	O	O
three	NN	O	O
internal	NN	O	O
peptides	NN	O	O
were	NN	O	O
sequenced	NN	O	O
.	NN	O	O

A	NN	O	O
comparison	NN	O	O
with	NN	O	O
known	NN	O	O
proteins	NN	O	O
in	NN	O	O
Genbank	NN	O	O
revealed	NN	O	O
that	NN	O	O
all	NN	O	O
three	NN	O	O
S6	NN	O	B-protein
peptides	NN	O	O
match	NN	O	O
the	NN	O	O
predicted	NN	O	O
sequence	NN	O	O
of	NN	O	O
TBP7	NN	O	B-protein
,	NN	O	O
Tat-binding	NN	O	B-protein
protein	NN	O	I-protein
7	NN	O	I-protein
.	NN	O	O

Based	NN	O	O
on	NN	O	O
peptide	NN	O	O
matches	NN	O	O
covering	NN	O	O
more	NN	O	O
than	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
TBP7	NN	O	B-protein
sequence	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
the	NN	O	O
migration	NN	O	O
of	NN	O	O
S6	NN	O	B-protein
on	NN	O	O
SDS-PAGE	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
estimated	NN	O	O
molecular	NN	O	O
mass	NN	O	O
for	NN	O	O
TBP7	NN	O	B-protein
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
subunit	NN	O	B-protein
6	NN	O	I-protein
of	NN	O	O
the	NN	O	O
26S	NN	O	B-protein
protease	NN	O	I-protein
is	NN	O	O
TBP7	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Hypoxia	NN	O	O
causes	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
through	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
on	NN	O	O
tyrosine	NN	O	O
residues	NN	O	O
.	NN	O	O

The	NN	O	O
response	NN	O	O
of	NN	O	O
mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
stress	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
such	NN	O	O
as	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
to	NN	O	O
induce	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
early	NN	O	B-DNA
response	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
exposure	NN	O	O
of	NN	O	O
cells	NN	O	O
to	NN	O	O
hypoxia	NN	O	O
(	NN	O	O
0.02	NN	O	O
%	NN	O	O
O2	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
degradation	NN	O	O
,	NN	O	O
increased	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
transactivation	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
construct	NN	O	I-DNA
containing	NN	O	O
two	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
DNA	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
protein	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
inhibitors	NN	O	O
and	NN	O	O
the	NN	O	O
dominant	NN	O	B-DNA
negative	NN	O	I-DNA
allele	NN	O	I-DNA
of	NN	O	O
c-Raf-1	NN	O	B-DNA
(	NN	O	O
Raf	NN	O	B-protein
301	NN	O	I-protein
)	NN	O	O
inhibited	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
degradation	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
,	NN	O	O
and	NN	O	O
transactivation	NN	O	O
of	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
by	NN	O	O
hypoxia	NN	O	O
.	NN	O	O

To	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
direct	NN	O	O
link	NN	O	O
between	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
pattern	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
with	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
we	NN	O	O
immunoprecipitated	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
after	NN	O	O
varying	NN	O	O
times	NN	O	O
of	NN	O	O
hypoxic	NN	O	O
exposure	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
its	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
status	NN	O	O
increased	NN	O	O
during	NN	O	O
hypoxic	NN	O	O
exposure	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
transfer	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphoryl	NN	O	O
groups	NN	O	O
onto	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
prevented	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
degradation	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
.	NN	O	O

In	NN	O	O
comparison	NN	O	O
to	NN	O	O
other	NN	O	O
activators	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
such	NN	O	O
as	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
or	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
we	NN	O	O
did	NN	O	O
not	NN	O	O
detect	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
status	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
following	NN	O	O
treatment	NN	O	O
with	NN	O	O
either	NN	O	O
of	NN	O	O
these	NN	O	O
agents	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
during	NN	O	O
hypoxia	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
proximal	NN	O	O
step	NN	O	O
which	NN	O	O
precedes	NN	O	O
its	NN	O	O
dissociation	NN	O	O
and	NN	O	O
degradation	NN	O	O
from	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Overproduction	NN	O	O
of	NN	O	O
NFKB2	NN	O	B-protein
(	NN	O	O
lyt-10	NN	O	B-protein
)	NN	O	O
and	NN	O	O
c-Rel	NN	O	B-protein
:	NN	O	O
a	NN	O	O
mechanism	NN	O	O
for	NN	O	O
HTLV-I	NN	O	B-protein
Tax	NN	O	I-protein
-mediated	NN	O	O
trans-activation	NN	O	O
via	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
signalling	NN	O	O
pathway	NN	O	O
.	NN	O	O

Molecular	NN	O	O
,	NN	O	O
biochemical	NN	O	O
and	NN	O	O
epidemiological	NN	O	O
evidence	NN	O	O
implicate	NN	O	O
HTLV-I	NN	O	O
as	NN	O	O
an	NN	O	O
etiologic	NN	O	O
agent	NN	O	O
of	NN	O	O
adult	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ATL	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
HTLV-I	NN	O	O
,	NN	O	O
a	NN	O	O
positive	NN	O	B-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	O
HTLV-I	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
viral	NN	O	O
oncogene	NN	O	O
that	NN	O	O
also	NN	O	O
increases	NN	O	O
transcription	NN	O	O
of	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
including	NN	O	O
GM-CSF	NN	O	B-protein
,	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	O
targets	NN	O	O
of	NN	O	O
the	NN	O	O
trans-activating	NN	O	O
effects	NN	O	O
of	NN	O	O
Tax	NN	O	O
is	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
pleiotropic	NN	O	B-protein
regulators	NN	O	I-protein
of	NN	O	O
immunoregulatory	NN	O	O
,	NN	O	O
cytokine	NN	O	O
and	NN	O	O
viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
NFKB2	NN	O	B-protein
(	NN	O	O
lyt-10	NN	O	B-protein
)	NN	O	O
and	NN	O	O
c-Rel	NN	O	B-protein
are	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
HTLV-I	NN	O	O
infected	NN	O	O
and	NN	O	O
Tax-expressing	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
,	NN	O	O
together	NN	O	O
,	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
PMA	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Most	NN	O	O
importantly	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
a	NN	O	O
Tax	NN	O	B-protein
-dependent	NN	O	O
correlation	NN	O	O
between	NN	O	O
expression	NN	O	O
of	NN	O	O
NFKB2	NN	O	B-protein
(	NN	O	O
p100	NN	O	B-protein
)	NN	O	O
and	NN	O	O
processing	NN	O	O
to	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
NFKB2	NN	O	B-protein
(	NN	O	O
p52	NN	O	B-protein
)	NN	O	O
form	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
and	NN	O	O
trans-activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-mediated	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
NFKB2	NN	O	B-protein
precursor	NN	O	O
is	NN	O	O
physically	NN	O	O
associated	NN	O	O
with	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	O
with	NN	O	O
Tax	NN	O	B-protein
in	NN	O	O
HTLV-I	NN	O	B-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
NFKB2	NN	O	B-protein
synthesis	NN	O	O
and	NN	O	O
processing	NN	O	O
allows	NN	O	O
continuous	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
otherwise	NN	O	O
cytoplasmic	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
,	NN	O	O
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
overexpression	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
NFKB2	NN	O	B-protein
alters	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
signalling	NN	O	O
pathway	NN	O	O
and	NN	O	O
contributes	NN	O	O
to	NN	O	O
leukemic	NN	O	O
transformation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

-DOCSTART-	O

Retinoic	NN	O	O
acid	NN	O	O
downmodulates	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
and	NN	O	O
GATA1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
purified	NN	O	B-cell_line
adult-progenitor	NN	O	I-cell_line
culture	NN	O	I-cell_line
.	NN	O	O

All-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
morphogen	NN	O	O
in	NN	O	O
vertebrate	NN	O	O
development	NN	O	O
,	NN	O	O
a	NN	O	O
normal	NN	O	O
constituent	NN	O	O
in	NN	O	O
human	NN	O	O
adult	NN	O	O
blood	NN	O	O
and	NN	O	O
is	NN	O	O
also	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
in	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
RA	NN	O	O
on	NN	O	O
normal	NN	O	O
hematopoiesis	NN	O	O
by	NN	O	O
using	NN	O	O
early	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HPC	NN	O	B-cell_type
)	NN	O	O
stringently	NN	O	O
purified	NN	O	O
from	NN	O	O
adult	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
.	NN	O	O

In	NN	O	O
clonogenetic	NN	O	B-cell_line
fetal	NN	O	I-cell_line
calf	NN	O	I-cell_line
serum-supplemented	NN	O	I-cell_line
(	NN	O	I-cell_line
FCS+	NN	O	I-cell_line
)	NN	O	I-cell_line
or	NN	O	I-cell_line
-nonsupplemented	NN	O	I-cell_line
(	NN	O	I-cell_line
FCS-	NN	O	I-cell_line
)	NN	O	I-cell_line
culture	NN	O	I-cell_line
treated	NN	O	O
with	NN	O	O
saturating	NN	O	O
levels	NN	O	O
of	NN	O	O
interleukin-3	NN	O	B-protein
(	NN	O	O
IL-3	NN	O	B-protein
)	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
Ep	NN	O	B-protein
)	NN	O	O
(	NN	O	O
combined	NN	O	O
with	NN	O	O
c-kit	NN	O	B-protein
ligand	NN	O	O
in	NN	O	O
FCS	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
-culture	NN	O	O
conditions	NN	O	O
)	NN	O	O
,	NN	O	O
RA	NN	O	O
induces	NN	O	O
a	NN	O	O
dramatic	NN	O	O
dose-dependent	NN	O	O
shift	NN	O	O
from	NN	O	O
erythroid	NN	O	O
to	NN	O	O
granulomonocytic	NN	O	O
colony	NN	O	O
formation	NN	O	O
,	NN	O	O
the	NN	O	O
latter	NN	O	O
colonies	NN	O	O
being	NN	O	O
essentially	NN	O	O
represented	NN	O	O
by	NN	O	O
granulocytic	NN	O	B-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
shift	NN	O	O
is	NN	O	O
apparently	NN	O	O
not	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
recruitment	NN	O	O
phenomenon	NN	O	O
,	NN	O	O
because	NN	O	O
in	NN	O	O
FCS+	NN	O	B-cell_line
culture	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
total	NN	O	O
number	NN	O	O
of	NN	O	O
colonies	NN	O	O
is	NN	O	O
not	NN	O	O
significantly	NN	O	O
modified	NN	O	O
by	NN	O	O
RA	NN	O	O
addition	NN	O	O
.	NN	O	O

In	NN	O	O
FCS-	NN	O	B-cell_line
liquid-suspension	NN	O	I-cell_line
culture	NN	O	I-cell_line
supplemented	NN	O	O
with	NN	O	O
saturating	NN	O	O
Ep	NN	O	B-protein
level	NN	O	O
and	NN	O	O
low-dose	NN	O	B-protein
IL-3/GM-CSF	NN	O	I-protein
,	NN	O	O
adult	NN	O	O
HPC	NN	O	B-cell_type
undergo	NN	O	O
unilineage	NN	O	O
erythropoietic	NN	O	O
differentiation	NN	O	O
:	NN	O	O
Here	NN	O	O
again	NN	O	O
,	NN	O	O
treatment	NN	O	O
with	NN	O	O
high-dose	NN	O	O
RA	NN	O	O
induces	NN	O	O
a	NN	O	O
shift	NN	O	O
from	NN	O	O
the	NN	O	O
erythroid	NN	O	O
to	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
pathway	NN	O	O
.	NN	O	O

Studies	NN	O	O
on	NN	O	O
RA	NN	O	O
time-response	NN	O	O
or	NN	O	O
pulse	NN	O	O
treatment	NN	O	O
in	NN	O	O
semisolid	NN	O	O
or	NN	O	O
liquid	NN	O	O
culture	NN	O	O
show	NN	O	O
that	NN	O	O
early	NN	O	O
RA	NN	O	O
addition	NN	O	O
is	NN	O	O
most	NN	O	O
effective	NN	O	O
,	NN	O	O
thus	NN	O	O
indicating	NN	O	O
that	NN	O	O
early	NN	O	O
but	NN	O	O
not	NN	O	O
late	NN	O	O
HPC	NN	O	B-cell_type
are	NN	O	O
sensitive	NN	O	O
to	NN	O	O
its	NN	O	O
action	NN	O	O
.	NN	O	O

We	NN	O	O
then	NN	O	O
analyzed	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
master	NN	O	B-DNA
GATA1	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
a	NN	O	O
finger	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
required	NN	O	O
for	NN	O	O
normal	NN	O	O
erythroid	NN	O	O
development	NN	O	O
;	NN	O	O
addition	NN	O	O
of	NN	O	O
RA	NN	O	O
to	NN	O	O
HPC	NN	O	B-cell_type
stimulated	NN	O	O
into	NN	O	O
unilineage	NN	O	O
erythropoietic	NN	O	O
differentiation	NN	O	O
in	NN	O	O
liquid	NN	O	O
culture	NN	O	O
caused	NN	O	O
a	NN	O	O
virtually	NN	O	O
complete	NN	O	O
inhibition	NN	O	O
of	NN	O	O
GATA1	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
RA	NN	O	O
directly	NN	O	O
inhibits	NN	O	O
the	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
program	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
early	NN	O	O
adult	NN	O	O
HPC	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
a	NN	O	O
shift	NN	O	O
from	NN	O	O
the	NN	O	O
erythroid	NN	O	O
to	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
pathway	NN	O	O
.	NN	O	O

This	NN	O	O
phenomenon	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
inhibition	NN	O	O
of	NN	O	O
GATA1	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
the	NN	O	O
early	NN	O	O
stages	NN	O	O
of	NN	O	O
erythropoietic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
phosphatidylinositol	NN	O	O
turnover	NN	O	O
and	NN	O	O
EGR-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
by	NN	O	O
crosslinking	NN	O	O
of	NN	O	O
surface	NN	O	O
IgM	NN	O	B-protein
and	NN	O	O
IgD	NN	O	B-protein
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
B104	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	I-cell_line
B104	NN	O	I-cell_line
,	NN	O	O
expressed	NN	O	O
surface	NN	O	B-protein
IgM	NN	O	I-protein
(	NN	O	O
sIgM	NN	O	B-protein
)	NN	O	O
and	NN	O	O
surface	NN	O	B-protein
IgD	NN	O	I-protein
(	NN	O	O
sIgD	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
crosslinking	NN	O	O
of	NN	O	O
sIgM	NN	O	B-protein
and	NN	O	O
sIgD	NN	O	B-protein
by	NN	O	O
anti-IgM	NN	O	B-protein
antibody	NN	O	I-protein
(	NN	O	I-protein
Ab	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
anti-IgD	NN	O	B-protein
Ab	NN	O	I-protein
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
induced	NN	O	O
Ca2+	NN	O	O
influx	NN	O	O
to	NN	O	O
almost	NN	O	O
the	NN	O	O
same	NN	O	O
degree	NN	O	O
,	NN	O	O
whereas	NN	O	O
only	NN	O	O
sIgM	NN	O	B-protein
-crosslinking	NN	O	O
caused	NN	O	O
B104	NN	O	B-cell_line
cell	NN	O	I-cell_line
death	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
(	NN	O	O
cAMP	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
hydrolysis	NN	O	O
of	NN	O	O
inositol	NN	O	O
phosphates	NN	O	O
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
Egr-1	NN	O	O
and	NN	O	O
c-fos	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
by	NN	O	O
sIgM-	NN	O	O
and	NN	O	O
sIgD-crosslinking	NN	O	O
to	NN	O	O
examine	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
signals	NN	O	O
mediated	NN	O	O
through	NN	O	O
sIgM	NN	O	B-protein
and	NN	O	O
sIgD	NN	O	B-protein
in	NN	O	O
B104	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Both	NN	O	O
sIgM-	NN	O	O
and	NN	O	O
sIgD-crosslinking	NN	O	O
with	NN	O	O
antibodies	NN	O	O
induced	NN	O	O
elevation	NN	O	O
of	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
,	NN	O	O
phosphatidylinositol	NN	O	O
turnover	NN	O	O
,	NN	O	O
PKC	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-RNA
and	NN	O	I-RNA
c-fos	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
although	NN	O	O
sIgM	NN	O	B-protein
-crosslinking	NN	O	O
was	NN	O	O
more	NN	O	O
effective	NN	O	O
than	NN	O	O
sIgD	NN	O	B-protein
-crosslinking	NN	O	O
,	NN	O	O
presumably	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
higher	NN	O	O
expression	NN	O	O
of	NN	O	O
sIgM	NN	O	B-protein
than	NN	O	O
of	NN	O	O
sIgD	NN	O	B-protein
.	NN	O	O

Egr-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
sIgM-	NN	O	O
and	NN	O	O
sIgD-crosslinking	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
H7	NN	O	B-protein
,	NN	O	O
erbstatin	NN	O	O
and	NN	O	O
genistein	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
HA1004	NN	O	O
.	NN	O	O

Erbstatin	NN	O	O
and	NN	O	O
genistein	NN	O	O
inhibited	NN	O	O
the	NN	O	O
sIg-crosslinking-induced	NN	O	O
Egr-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
parallel	NN	O	O
to	NN	O	O
that	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
sIg-crosslinking-induced	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

Phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
induced	NN	O	O
Egr-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
but	NN	O	O
forskolin	NN	O	O
and	NN	O	O
dibutyryl	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
Egr-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
activating	NN	O	O
signals	NN	O	O
through	NN	O	O
sIgM	NN	O	B-protein
and	NN	O	O
sIgD	NN	O	B-protein
are	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
kinase-	NN	O	O
and	NN	O	O
PKC-dependent	NN	O	O
,	NN	O	O
but	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
-independent	NN	O	O
.	NN	O	O

Cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
and	NN	O	O
FK506	NN	O	O
rescued	NN	O	O
B104	NN	O	B-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
death	NN	O	O
induced	NN	O	O
by	NN	O	O
anti-IgM	NN	O	B-protein
Ab	NN	O	I-protein
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
and	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
showing	NN	O	O
that	NN	O	O
CsA	NN	O	O
and	NN	O	O
FK506	NN	O	O
affect	NN	O	O
signal	NN	O	O
transducers	NN	O	O
differently	NN	O	O
from	NN	O	O
or	NN	O	O
downstream	NN	O	O
to	NN	O	O
these	NN	O	O
molecules	NN	O	O
.	NN	O	O

The	NN	O	O
difference	NN	O	O
in	NN	O	O
signals	NN	O	O
transduced	NN	O	O
through	NN	O	O
sIgM	NN	O	B-protein
and	NN	O	O
sIgD	NN	O	B-protein
in	NN	O	O
B104	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Direct	NN	O	O
exposure	NN	O	O
to	NN	O	O
2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8-tetrachlorodibenzo-p-dioxin	NN	O	O
(	NN	O	O
TCDD	NN	O	O
)	NN	O	O
increases	NN	O	O
infectivity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
to	NN	O	O
a	NN	O	O
malarial	NN	O	O
parasite	NN	O	O
.	NN	O	O

Direct	NN	O	O
exposure	NN	O	O
to	NN	O	O
10	NN	O	O
nM	NN	O	O
2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8-TCDD	NN	O	O
caused	NN	O	O
a	NN	O	O
75	NN	O	O
%	NN	O	O
increase	NN	O	O
and	NN	O	O
a	NN	O	O
2-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
infectivity	NN	O	O
of	NN	O	O
isolated	NN	O	O
human	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
to	NN	O	O
P.	NN	O	O
falciparum	NN	O	O
after	NN	O	O
48	NN	O	O
hours	NN	O	O
when	NN	O	O
the	NN	O	O
parasites	NN	O	O
were	NN	O	O
in	NN	O	O
an	NN	O	O
unsynchronized	NN	O	O
or	NN	O	O
synchronized	NN	O	O
state	NN	O	O
of	NN	O	O
growth	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
with	NN	O	O
10	NN	O	O
microM	NN	O	O
sodium	NN	O	O
orthovanadate	NN	O	O
(	NN	O	O
NaOV	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
plasma	NN	O	B-protein
membrane	NN	O	I-protein
Ca-ATPase	NN	O	I-protein
and	NN	O	O
phosphotyrosine	NN	O	B-protein
phosphatase	NN	O	I-protein
,	NN	O	O
decreased	NN	O	O
parasitemia	NN	O	O
by	NN	O	O
30	NN	O	O
%	NN	O	O
.	NN	O	O

Co-treatment	NN	O	O
of	NN	O	O
RBCs	NN	O	B-cell_type
with	NN	O	O
TCDD	NN	O	O
and	NN	O	O
NaOV	NN	O	O
completely	NN	O	O
blocked	NN	O	O
the	NN	O	O
TCDD-induced	NN	O	O
increase	NN	O	O
in	NN	O	O
parasitemia	NN	O	O
.	NN	O	O

Because	NN	O	O
erythrocytes	NN	O	B-cell_type
are	NN	O	O
anucleated	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
are	NN	O	O
discussed	NN	O	O
as	NN	O	O
evidence	NN	O	O
for	NN	O	O
biochemical	NN	O	O
changes	NN	O	O
by	NN	O	O
TCDD	NN	O	O
without	NN	O	O
requiring	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
gene	NN	O	B-protein
products	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Evidence	NN	O	O
for	NN	O	O
a	NN	O	O
trans-acting	NN	O	B-protein
activator	NN	O	I-protein
function	NN	O	O
regulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
CD5	NN	O	I-protein
antigen	NN	O	I-protein
.	NN	O	O

Interspecies	NN	O	O
somatic	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybrids	NN	O	I-cell_line
were	NN	O	O
generated	NN	O	O
by	NN	O	O
fusing	NN	O	O
the	NN	O	O
mouse	NN	O	B-cell_line
T-lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
BW5147	NN	O	B-cell_line
,	NN	O	O
with	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
at	NN	O	O
different	NN	O	O
stages	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Thymocytes	NN	O	B-cell_type
,	NN	O	O
activated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
or	NN	O	O
an	NN	O	O
activated	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
were	NN	O	O
used	NN	O	O
as	NN	O	O
human	NN	O	O
partners	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
in	NN	O	O
three	NN	O	O
independent	NN	O	O
fusions	NN	O	O
.	NN	O	O

Irrespective	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
cell	NN	O	O
partner	NN	O	O
used	NN	O	O
for	NN	O	O
fusion	NN	O	O
,	NN	O	O
a	NN	O	O
certain	NN	O	O
number	NN	O	O
of	NN	O	O
hybrids	NN	O	B-cell_line
lost	NN	O	O
CD5	NN	O	B-protein
surface	NN	O	O
expression	NN	O	O
over	NN	O	O
a	NN	O	O
period	NN	O	O
of	NN	O	O
time	NN	O	O
in	NN	O	O
culture	NN	O	O
.	NN	O	O

Analysis	NN	O	O
at	NN	O	O
the	NN	O	O
phenotype	NN	O	O
and	NN	O	O
genetic	NN	O	O
level	NN	O	O
showed	NN	O	O
that	NN	O	O
lack	NN	O	O
of	NN	O	O
CD5	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
due	NN	O	O
neither	NN	O	O
to	NN	O	O
segregation	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
autosome	NN	O	I-DNA
11	NN	O	I-DNA
,	NN	O	O
on	NN	O	O
which	NN	O	O
the	NN	O	O
CD5	NN	O	B-DNA
gene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
,	NN	O	O
nor	NN	O	O
to	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
CD5	NN	O	B-DNA
structural	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
loss	NN	O	O
of	NN	O	O
CD5	NN	O	B-protein
surface	NN	O	O
expression	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
specific	NN	O	O
mRNA	NN	O	B-RNA
.	NN	O	O

Since	NN	O	O
these	NN	O	O
hybrids	NN	O	O
preferentially	NN	O	O
segregate	NN	O	O
human	NN	O	O
chromosomes	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
indicate	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
non-syntenic	NN	O	B-DNA
trans-active	NN	O	I-DNA
locus	NN	O	I-DNA
,	NN	O	O
or	NN	O	O
loci	NN	O	O
,	NN	O	O
positively	NN	O	O
controlling	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD5	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
gene	NN	O	I-DNA
during	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U937	NN	O	I-cell_line
requires	NN	O	O
a	NN	O	O
novel	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
MS-2	NN	O	B-protein
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
J	NN	O	O
Immunol	NN	O	O
1999	NN	O	O
Jul	NN	O	O
15	NN	O	O
;	NN	O	O
163	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
:	NN	O	O
1091	NN	O	O
]	NN	O	O

The	NN	O	O
differentiation	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
precursors	NN	O	I-cell_type
into	NN	O	O
mature	NN	O	O
myelomonocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
beta2	NN	O	B-protein
integrin	NN	O	I-protein
CD11b	NN	O	B-protein
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
Sp1	NN	O	B-protein
and	NN	O	O
PU.1	NN	O	B-protein
prime	NN	O	I-protein
the	NN	O	O
CD11b	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
factors	NN	O	O
responsible	NN	O	O
for	NN	O	O
its	NN	O	O
inducible	NN	O	O
expression	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
homologous	NN	O	O
genes	NN	O	O
encoding	NN	O	O
CD11a	NN	O	B-protein
and	NN	O	O
CD11c	NN	O	B-protein
also	NN	O	O
exhibit	NN	O	O
inducible	NN	O	O
expression	NN	O	O
during	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
compared	NN	O	O
the	NN	O	O
nucleotide	NN	O	B-DNA
sequences	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CD11a	NN	O	B-DNA
,	NN	O	I-DNA
CD11b	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
CD11c	NN	O	I-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
to	NN	O	O
identify	NN	O	O
common	NN	O	O
elements	NN	O	O
that	NN	O	O
might	NN	O	O
contribute	NN	O	O
to	NN	O	O
inducible	NN	O	O
expression	NN	O	O
.	NN	O	O

This	NN	O	O
analysis	NN	O	O
identified	NN	O	O
one	NN	O	O
such	NN	O	O
element	NN	O	O
repeated	NN	O	O
four	NN	O	O
times	NN	O	O
within	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
these	NN	O	O
elements	NN	O	O
indicated	NN	O	O
that	NN	O	O
two	NN	O	O
,	NN	O	O
MS-2beta	NN	O	B-protein
and	NN	O	O
MS-2gamma	NN	O	B-protein
,	NN	O	O
are	NN	O	O
critical	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
gene	NN	O	I-DNA
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
pro-monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U937	NN	O	I-cell_line
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
indicate	NN	O	O
that	NN	O	O
MS-2beta	NN	O	B-protein
and	NN	O	O
MS-2gamma	NN	O	B-protein
interact	NN	O	O
with	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
induced	NN	O	O
during	NN	O	O
U937	NN	O	B-cell_line
differentiation	NN	O	O
.	NN	O	O

These	NN	O	O
factors	NN	O	O
are	NN	O	O
detected	NN	O	O
at	NN	O	O
the	NN	O	O
time	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
activated	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
is	NN	O	O
approximately	NN	O	O
28	NN	O	O
kDa	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
DNA	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
are	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
novel	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
MS-2	NN	O	B-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
MS-2	NN	O	B-protein
mediates	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
gene	NN	O	I-DNA
as	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
monocytic	NN	O	O
lineage	NN	O	O
mature	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
multiple	NN	O	O
potential	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
MS-2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
promoter	NN	O	O
regions	NN	O	O
of	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
genes	NN	O	O
expressed	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
suggests	NN	O	O
this	NN	O	O
factor	NN	O	O
plays	NN	O	O
a	NN	O	O
general	NN	O	O
role	NN	O	O
in	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Acetylsalicylic	NN	O	O
acid	NN	O	O
and	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
inhibit	NN	O	O
LPS-induced	NN	O	O
NF-kappa	NN	O	B-protein
B/c-Rel	NN	O	I-protein
nuclear	NN	O	O
translocation	NN	O	O
,	NN	O	O
and	NN	O	O
synthesis	NN	O	O
of	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
TF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alfa	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
acetylsalicylic	NN	O	O
acid	NN	O	O
and	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
on	NN	O	O
the	NN	O	O
LPS-induced	NN	O	O
synthesis	NN	O	O
of	NN	O	O
the	NN	O	O
pro-coagulant	NN	O	O
protein	NN	O	O
tissue	NN	O	O
factor	NN	O	O
(	NN	O	O
TF	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
pro-inflammatory	NN	O	O
protein	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor-alpha	NN	O	O
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
prostaglandin	NN	O	B-protein
PGE2	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Both	NN	O	O
drugs	NN	O	O
dose-dependently	NN	O	O
inhibited	NN	O	O
LPS-induced	NN	O	B-protein
TF	NN	O	I-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
synthesis	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
and	NN	O	O
the	NN	O	O
protein	NN	O	O
level	NN	O	O
,	NN	O	O
and	NN	O	O
reduced	NN	O	O
PGE2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

As	NN	O	O
evidenced	NN	O	O
by	NN	O	O
electro	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
prototypic	NN	O	I-DNA
probe	NN	O	I-DNA
,	NN	O	O
these	NN	O	O
drugs	NN	O	O
probably	NN	O	O
exert	NN	O	O
their	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
by	NN	O	O
interference	NN	O	O
with	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/c-Rel	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
may	NN	O	O
expand	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
anti-thrombotic	NN	O	O
and	NN	O	O
anti-inflammatory	NN	O	O
effects	NN	O	O
of	NN	O	O
these	NN	O	O
drugs	NN	O	O
when	NN	O	O
activation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
occurs	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interferon	NN	O	O
augments	NN	O	O
PML	NN	O	B-protein
and	NN	O	O
PML/RAR	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
normal	NN	O	O
myeloid	NN	O	B-cell_type
and	NN	O	O
acute	NN	O	B-cell_type
promyelocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
cooperates	NN	O	O
with	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
to	NN	O	O
induce	NN	O	O
maturation	NN	O	O
of	NN	O	O
a	NN	O	O
retinoid-resistant	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
PML	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
fused	NN	O	O
to	NN	O	O
the	NN	O	O
retinoic	NN	O	B-DNA
acid	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
RAR	NN	O	B-DNA
alpha	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
15	NN	O	O
;	NN	O	O
17	NN	O	O
translocation	NN	O	O
.	NN	O	O

PML	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
diverse	NN	O	O
tissues	NN	O	O
and	NN	O	O
cell	NN	O	O
lines	NN	O	O
and	NN	O	O
localized	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
with	NN	O	O
a	NN	O	O
typical	NN	O	O
speckled	NN	O	O
pattern	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
bone	NN	O	O
marrow	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
preferentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

PML	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
transcriptionally	NN	O	O
regulated	NN	O	O
by	NN	O	O
class	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
II	NN	O	I-protein
interferons	NN	O	I-protein
,	NN	O	O
which	NN	O	O
raises	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
interferons	NN	O	O
modulate	NN	O	O
the	NN	O	O
function	NN	O	O
and	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
potential	NN	O	O
of	NN	O	O
normal	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
precursors	NN	O	O
by	NN	O	O
activating	NN	O	O
PML	NN	O	B-protein
-dependent	NN	O	O
pathways	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
interferons	NN	O	O
could	NN	O	O
act	NN	O	O
on	NN	O	O
APL	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
alone	NN	O	O
or	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
,	NN	O	O
especially	NN	O	O
if	NN	O	O
the	NN	O	O
PML/RAR	NN	O	B-RNA
alpha	NN	O	I-RNA
fusion	NN	O	I-RNA
transcript	NN	O	I-RNA
that	NN	O	O
results	NN	O	O
from	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
15	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
is	NN	O	O
induced	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
PML	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
low	NN	O	O
levels	NN	O	O
or	NN	O	O
not	NN	O	O
expressed	NN	O	O
in	NN	O	O
normal	NN	O	O
circulating	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
polymorphonucleate	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
is	NN	O	O
markedly	NN	O	O
induced	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
;	NN	O	O
that	NN	O	O
PML	NN	O	B-protein
and	NN	O	O
PML/RAR	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
is	NN	O	O
augmented	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
in	NN	O	O
the	NN	O	O
NB4	NN	O	B-cell_line
APL	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
carries	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
15	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
in	NN	O	O
APL	NN	O	B-cell_type
blasts	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
;	NN	O	O
that	NN	O	O
interferon	NN	O	B-protein
inhibits	NN	O	O
growth	NN	O	O
and	NN	O	O
survival	NN	O	O
of	NN	O	O
NB4	NN	O	O
APL	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
cooperation	NN	O	O
with	NN	O	O
RA	NN	O	O
;	NN	O	O
that	NN	O	O
interferons	NN	O	O
alone	NN	O	O
have	NN	O	O
minimal	NN	O	O
maturation	NN	O	O
effect	NN	O	O
on	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
;	NN	O	O
and	NN	O	O
,	NN	O	O
finally	NN	O	O
,	NN	O	O
that	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
,	NN	O	I-protein
but	NN	O	I-protein
not	NN	O	I-protein
alpha	NN	O	I-protein
or	NN	O	I-protein
beta	NN	O	I-protein
,	NN	O	O
induces	NN	O	O
maturation	NN	O	O
and	NN	O	O
growth	NN	O	O
suppression	NN	O	O
of	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
de	NN	O	O
novo	NN	O	O
retinoid	NN	O	O
resistance	NN	O	O
,	NN	O	O
and	NN	O	O
partially	NN	O	O
restores	NN	O	O
RA	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
Ara-C	NN	O	O
on	NN	O	O
neutral	NN	O	B-protein
sphingomyelinase	NN	O	I-protein
and	NN	O	O
mitogen-	NN	O	B-protein
and	NN	O	I-protein
stress-	NN	O	I-protein
activated	NN	O	I-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
in	NN	O	O
T-lymphocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Neutral	NN	O	B-protein
sphingomyelinase	NN	O	I-protein
(	NN	O	O
SMase	NN	O	B-protein
)	NN	O	O
can	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
extracellular	NN	O	O
signals	NN	O	O
to	NN	O	O
produce	NN	O	O
ceramide	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
affect	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
activities	NN	O	O
.	NN	O	O

Neutral	NN	O	O
SMase	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
assessed	NN	O	O
in	NN	O	O
membranes	NN	O	O
from	NN	O	O
Jurkat	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
EL4	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
murine	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Ara-C	NN	O	O
activated	NN	O	O
SMase	NN	O	B-protein
with	NN	O	O
10	NN	O	O
minutes	NN	O	O
in	NN	O	O
both	NN	O	O
Jurkat	NN	O	O
and	NN	O	O
EL4	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

PMA	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
Ara-C	NN	O	O
or	NN	O	O
ceramides	NN	O	O
,	NN	O	O
activated	NN	O	O
ERK	NN	O	B-protein
MAPKS	NN	O	I-protein
,	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
and	NN	O	O
EL4	NN	O	B-cell_line
.	NN	O	O

PMA	NN	O	O
acted	NN	O	O
synergistically	NN	O	O
with	NN	O	O
ionomycin	NN	O	O
to	NN	O	O
activate	NN	O	O
JNK	NN	O	B-protein
MAPKs	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
and	NN	O	O
EL4	NN	O	B-cell_line
within	NN	O	O
10	NN	O	O
minutes	NN	O	O
.	NN	O	O

Ara-C	NN	O	O
activated	NN	O	O
JNKs	NN	O	B-protein
only	NN	O	O
after	NN	O	O
prolonged	NN	O	O
incubation	NN	O	O
(	NN	O	O
90-120	NN	O	O
minutes	NN	O	O
)	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
ceramide	NN	O	O
is	NN	O	O
not	NN	O	O
a	NN	O	O
positive	NN	O	O
signal	NN	O	O
for	NN	O	O
ERK	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
Ara-C	NN	O	O
on	NN	O	O
JNK	NN	O	B-protein
activity	NN	O	O
may	NN	O	O
be	NN	O	O
mediated	NN	O	O
through	NN	O	O
secondary	NN	O	O
response	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

Comparative	NN	O	O
analysis	NN	O	O
identifies	NN	O	O
conserved	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
receptor-associated	NN	O	I-DNA
factor	NN	O	I-DNA
3	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
simian	NN	O	I-DNA
Epstein-Barr	NN	O	I-DNA
virus	NN	O	I-DNA
oncogene	NN	O	I-DNA
LMP1	NN	O	I-DNA
.	NN	O	O

Nonhuman	NN	O	O
primates	NN	O	O
are	NN	O	O
naturally	NN	O	O
infected	NN	O	O
with	NN	O	O
a	NN	O	O
B-lymphotropic	NN	O	O
herpesvirus	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
simian	NN	O	O
EBV	NN	O	O
share	NN	O	O
considerable	NN	O	O
genetic	NN	O	O
,	NN	O	O
biologic	NN	O	O
,	NN	O	O
and	NN	O	O
epidemiologic	NN	O	O
features	NN	O	O
with	NN	O	O
human	NN	O	O
EBV	NN	O	O
,	NN	O	O
including	NN	O	O
virus-induced	NN	O	O
tumorigenesis	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
latent	NN	O	O
,	NN	O	O
transformation-associated	NN	O	O
viral	NN	O	O
genes	NN	O	O
demonstrate	NN	O	O
marked	NN	O	O
sequence	NN	O	O
divergence	NN	O	O
among	NN	O	O
species	NN	O	O
despite	NN	O	O
the	NN	O	O
conserved	NN	O	O
functions	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
the	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
LMP1	NN	O	I-protein
)	NN	O	I-protein
homologs	NN	O	I-protein
from	NN	O	O
the	NN	O	O
simian	NN	O	O
EBV	NN	O	O
naturally	NN	O	O
infecting	NN	O	O
baboons	NN	O	O
(	NN	O	O
cercopithicine	NN	O	O
herpesvirus	NN	O	O
12	NN	O	O
,	NN	O	O
herpesvirus	NN	O	O
papio	NN	O	O
)	NN	O	O
and	NN	O	O
rhesus	NN	O	O
monkeys	NN	O	O
(	NN	O	O
cercopithicine	NN	O	O
herpesvirus	NN	O	O
15	NN	O	O
)	NN	O	O
for	NN	O	O
a	NN	O	O
comparative	NN	O	O
study	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	O
EBV	NN	O	O
oncogene	NN	O	O
.	NN	O	O

The	NN	O	O
transmembrane	NN	O	O
domains	NN	O	O
are	NN	O	O
well	NN	O	O
conserved	NN	O	O
,	NN	O	O
but	NN	O	O
there	NN	O	O
is	NN	O	O
striking	NN	O	O
sequence	NN	O	O
divergence	NN	O	O
of	NN	O	O
the	NN	O	O
carboxy-terminal	NN	O	B-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
essential	NN	O	O
for	NN	O	O
B-cell	NN	O	B-cell_type
immortalization	NN	O	O
and	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
the	NN	O	O
simian	NN	O	B-protein
EBV	NN	O	I-protein
LMP1s	NN	O	I-protein
retain	NN	O	O
most	NN	O	O
functions	NN	O	O
in	NN	O	O
common	NN	O	O
with	NN	O	O
EBV	NN	O	O
LMP1	NN	O	B-protein
,	NN	O	O
including	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
NF-	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
to	NN	O	O
bind	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-associated	NN	O	I-protein
factor	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
TRAF3	NN	O	B-protein
)	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor-responsive	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
ICAM1	NN	O	B-DNA
,	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Multiple	NN	O	O
TRAF3	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
containing	NN	O	O
a	NN	O	O
PXQXT/S	NN	O	B-DNA
core	NN	O	I-DNA
sequence	NN	O	I-DNA
can	NN	O	O
be	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
simian	NN	O	B-protein
EBV	NN	O	I-protein
LMP1s	NN	O	I-protein
by	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
binding	NN	O	O
assay	NN	O	O
.	NN	O	O

A	NN	O	O
PXQXT/S-containing	NN	O	B-DNA
sequence	NN	O	I-DNA
is	NN	O	O
also	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
marker	NN	O	O
,	NN	O	O
CD30	NN	O	B-protein
,	NN	O	O
and	NN	O	O
binds	NN	O	O
TRAF3	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
last	NN	O	B-protein
13	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
containing	NN	O	O
a	NN	O	O
PXQXT/S	NN	O	B-DNA
sequence	NN	O	I-DNA
are	NN	O	O
highly	NN	O	O
conserved	NN	O	O
in	NN	O	O
human	NN	O	O
and	NN	O	O
simian	NN	O	O
EBV	NN	O	O
LMP1	NN	O	B-protein
but	NN	O	O
do	NN	O	O
not	NN	O	O
bind	NN	O	O
TRAF3	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
distinct	NN	O	O
role	NN	O	O
for	NN	O	O
this	NN	O	O
conserved	NN	O	O
region	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
conserved	NN	O	O
TRAF3	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
LMP1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
CD30	NN	O	B-protein
Hodgkin	NN	O	I-protein
's	NN	O	I-protein
disease	NN	O	I-protein
marker	NN	O	I-protein
provides	NN	O	O
further	NN	O	O
evidence	NN	O	O
that	NN	O	O
a	NN	O	O
TRAF3	NN	O	B-protein
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
malignant	NN	O	O
transformation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Chromosome	NN	O	B-DNA
1	NN	O	I-DNA
aneusomy	NN	O	O
with	NN	O	O
1p36	NN	O	B-DNA
under-representation	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
histologic	NN	O	O
grade	NN	O	O
,	NN	O	O
DNA	NN	O	O
aneuploidy	NN	O	O
,	NN	O	O
high	NN	O	O
c-erb	NN	O	B-protein
B-2	NN	O	I-protein
and	NN	O	O
loss	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
ductal	NN	O	O
breast	NN	O	O
carcinoma	NN	O	O
.	NN	O	O

Chromosome	NN	O	B-DNA
1	NN	O	I-DNA
abnormalities	NN	O	O
with	NN	O	O
loss	NN	O	O
of	NN	O	O
1p36	NN	O	B-DNA
have	NN	O	O
been	NN	O	O
investigated	NN	O	O
in	NN	O	O
95	NN	O	O
breast-cancer	NN	O	O
samples	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
a	NN	O	O
dual-target	NN	O	O
fluorescence	NN	O	O
in-situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
technique	NN	O	O
using	NN	O	O
the	NN	O	O
pUC	NN	O	B-DNA
1.77	NN	O	I-DNA
and	NN	O	O
p1-79	NN	O	B-DNA
probes	NN	O	I-DNA
,	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
1q12	NN	O	B-DNA
and	NN	O	I-DNA
1p36	NN	O	I-DNA
regions	NN	O	I-DNA
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Abnormalities	NN	O	O
for	NN	O	O
one	NN	O	O
or	NN	O	O
both	NN	O	O
probes	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
83/95	NN	O	O
samples	NN	O	O
.	NN	O	O

Relative	NN	O	O
1p36	NN	O	B-DNA
under-representation	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
79/95	NN	O	O
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
relevance	NN	O	O
of	NN	O	O
these	NN	O	O
alterations	NN	O	O
was	NN	O	O
studied	NN	O	O
by	NN	O	O
comparing	NN	O	O
the	NN	O	O
FISH	NN	O	O
results	NN	O	O
with	NN	O	O
several	NN	O	O
parameters	NN	O	O
currently	NN	O	O
used	NN	O	O
in	NN	O	O
breast-cancer	NN	O	O
pathology	NN	O	O
.	NN	O	O

Distinct	NN	O	O
patterns	NN	O	O
of	NN	O	O
chromosome	NN	O	B-DNA
1	NN	O	I-DNA
abnormalities	NN	O	O
were	NN	O	O
found	NN	O	O
among	NN	O	O
the	NN	O	O
histologic	NN	O	O
types	NN	O	O
of	NN	O	O
breast	NN	O	O
carcinoma	NN	O	O
.	NN	O	O

Lobular	NN	O	O
or	NN	O	O
mucinous	NN	O	O
samples	NN	O	O
showed	NN	O	O
few	NN	O	O
or	NN	O	O
no	NN	O	O
alterations	NN	O	O
,	NN	O	O
whereas	NN	O	O
most	NN	O	O
ductal	NN	O	O
samples	NN	O	O
had	NN	O	O
high	NN	O	O
chromosome	NN	O	B-DNA
1	NN	O	I-DNA
polysomy	NN	O	O
with	NN	O	O
under-representation	NN	O	O
of	NN	O	O
1p36	NN	O	B-DNA
.	NN	O	O

In	NN	O	O
ductal	NN	O	O
carcinomas	NN	O	O
,	NN	O	O
chromosome	NN	O	B-DNA
1	NN	O	I-DNA
alterations	NN	O	O
increased	NN	O	O
with	NN	O	O
histologic	NN	O	O
grade	NN	O	O
,	NN	O	O
DNA	NN	O	O
aneuploidy	NN	O	O
,	NN	O	O
loss	NN	O	O
of	NN	O	O
bcl-2	NN	O	O
and	NN	O	O
high	NN	O	O
c-erb	NN	O	B-protein
B-2	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

These	NN	O	O
associations	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
statistically	NN	O	O
significant	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
between	NN	O	O
chromosome	NN	O	B-DNA
1	NN	O	I-DNA
alterations	NN	O	O
and	NN	O	O
nuclear	NN	O	O
grade	NN	O	O
,	NN	O	O
age	NN	O	O
,	NN	O	O
size	NN	O	O
,	NN	O	O
lymph-node	NN	O	O
involvement	NN	O	O
,	NN	O	O
hormonal	NN	O	O
receptor	NN	O	O
presence	NN	O	O
,	NN	O	O
proliferation	NN	O	O
activity	NN	O	O
or	NN	O	O
p53	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
was	NN	O	O
detected	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
the	NN	O	O
utility	NN	O	O
of	NN	O	O
this	NN	O	O
FISH	NN	O	O
technique	NN	O	O
for	NN	O	O
a	NN	O	O
better	NN	O	O
definition	NN	O	O
of	NN	O	O
the	NN	O	O
biological	NN	O	O
characteristics	NN	O	O
of	NN	O	O
ductal	NN	O	O
carcinomas	NN	O	O
.	NN	O	O

-DOCSTART-	O

G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra	NN	O	O
,	NN	O	O
a	NN	O	O
natural	NN	O	O
bacterial	NN	O	O
cell	NN	O	O
wall	NN	O	O
breakdown	NN	O	O
product	NN	O	O
,	NN	O	O
induces	NN	O	O
interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
interleukin-6	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
study	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
inflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
expression	NN	O	O
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
J	NN	O	O
Biol	NN	O	O
Chem	NN	O	O
1994	NN	O	O
Jun	NN	O	O
17	NN	O	O
;	NN	O	O
269	NN	O	O
(	NN	O	O
24	NN	O	O
)	NN	O	O
:	NN	O	O
16983	NN	O	O
]	NN	O	O

It	NN	O	O
is	NN	O	O
believed	NN	O	O
that	NN	O	O
induction	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
bacterial	NN	O	O
cell	NN	O	O
wall	NN	O	O
components	NN	O	O
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
and	NN	O	O
course	NN	O	O
of	NN	O	O
sepsis	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
most	NN	O	O
attention	NN	O	O
has	NN	O	O
been	NN	O	O
focused	NN	O	O
on	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
N-acetylglucosaminyl-1	NN	O	O
,	NN	O	O
6-anhydro-N-acetylmuramyl-L-alanyl-D-	NN	O	O
isoglutamyl-m-diaminopimelyl-D-alanine	NN	O	O
(	NN	O	O
G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
breakdown	NN	O	O
product	NN	O	O
of	NN	O	O
peptidoglycan	NN	O	O
that	NN	O	O
is	NN	O	O
produced	NN	O	O
by	NN	O	O
soluble	NN	O	O
lytic	NN	O	B-protein
transglycosylase	NN	O	I-protein
of	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
,	NN	O	O
to	NN	O	O
induce	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
strongly	NN	O	O
induce	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
beta	NN	O	I-protein
and	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
after	NN	O	O
2	NN	O	O
h	NN	O	O
and	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
IL-6	NN	O	B-protein
protein	NN	O	I-protein
secretion	NN	O	O
after	NN	O	O
48	NN	O	O
h	NN	O	O
of	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
increase	NN	O	O
in	NN	O	O
mRNA	NN	O	O
accumulation	NN	O	O
was	NN	O	O
at	NN	O	O
least	NN	O	O
partly	NN	O	O
due	NN	O	O
to	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
rates	NN	O	O
of	NN	O	O
the	NN	O	O
respective	NN	O	O
genes	NN	O	B-DNA
and	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
strong	NN	O	O
induction	NN	O	O
of	NN	O	O
nuclear	NN	O	O
factor-kappa	NN	O	O
B	NN	O	O
and	NN	O	O
activator	NN	O	O
protein-1	NN	O	O
transcription	NN	O	O
factor	NN	O	O
expression	NN	O	O
.	NN	O	O

Experiments	NN	O	O
using	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
,	NN	O	O
and	NN	O	O
tyrosine	NN	O	O
kinase-dependent	NN	O	O
pathways	NN	O	O
revealed	NN	O	O
that	NN	O	O
G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra-induced	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
involves	NN	O	O
activation	NN	O	O
of	NN	O	O
an	NN	O	O
H7-inhibitable	NN	O	O
pathway	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
the	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
inhibitor	NN	O	O
cycloheximide	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
shown	NN	O	O
that	NN	O	O
G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra-induced	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
new	NN	O	O
protein	NN	O	O
,	NN	O	O
whereas	NN	O	O
G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra-induced	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
mRNA	NN	O	O
accumulation	NN	O	O
does	NN	O	O
not	NN	O	O
.	NN	O	O

When	NN	O	O
responses	NN	O	O
to	NN	O	O
G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra	NN	O	O
were	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
to	NN	O	O
LPS	NN	O	O
and	NN	O	O
muramyldipeptide	NN	O	O
(	NN	O	O
MDP	NN	O	O
)	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
optimal	NN	O	O
response	NN	O	O
to	NN	O	O
G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra	NN	O	O
induction	NN	O	O
was	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
LPS	NN	O	O
but	NN	O	O
significantly	NN	O	O
higher	NN	O	O
than	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
MDP	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
maximal	NN	O	O
G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra-induced	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
could	NN	O	O
be	NN	O	O
enhanced	NN	O	O
by	NN	O	O
co-stimulation	NN	O	O
with	NN	O	O
LPS	NN	O	O
or	NN	O	O
MDP	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
different	NN	O	O
receptors	NN	O	O
and/or	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
were	NN	O	O
involved	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra	NN	O	O
induces	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
IL-6	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
suggesting	NN	O	O
a	NN	O	O
possible	NN	O	O
role	NN	O	O
for	NN	O	O
G	NN	O	O
(	NN	O	O
Anh	NN	O	O
)	NN	O	O
MTetra	NN	O	O
in	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
during	NN	O	O
sepsis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Increased	NN	O	O
proliferation	NN	O	O
,	NN	O	O
cytotoxicity	NN	O	O
,	NN	O	O
and	NN	O	O
gene	NN	O	O
expression	NN	O	O
after	NN	O	O
stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
through	NN	O	O
a	NN	O	O
surface	NN	O	O
ganglioside	NN	O	O
(	NN	O	O
GD3	NN	O	O
)	NN	O	O
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
J	NN	O	O
Immunol	NN	O	O
1994	NN	O	O
Jul	NN	O	O
15	NN	O	O
;	NN	O	O
153	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
:	NN	O	O
910	NN	O	O
]	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
gangliosides	NN	O	O
have	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
cell	NN	O	O
signaling	NN	O	O
and	NN	O	O
recognition	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
their	NN	O	O
specific	NN	O	O
function	NN	O	O
in	NN	O	O
these	NN	O	O
processes	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
clearly	NN	O	O
defined	NN	O	O
.	NN	O	O

A	NN	O	O
mAb	NN	O	B-protein
,	NN	O	O
R24	NN	O	B-protein
,	NN	O	O
that	NN	O	O
reacts	NN	O	O
specifically	NN	O	O
with	NN	O	O
a	NN	O	O
cell	NN	O	O
surface	NN	O	O
ganglioside	NN	O	O
(	NN	O	O
GD3	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
stimulate	NN	O	O
proliferation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
derived	NN	O	O
from	NN	O	O
human	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
the	NN	O	O
R24	NN	O	B-protein
mAb	NN	O	I-protein
affects	NN	O	O
T	NN	O	O
cell	NN	O	O
functions	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
observed	NN	O	O
that	NN	O	O
the	NN	O	O
R24	NN	O	B-protein
mAb	NN	O	I-protein
stimulates	NN	O	O
GD3+	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
cytotoxicity	NN	O	O
,	NN	O	O
and	NN	O	O
surface	NN	O	O
marker	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
alpha-chain	NN	O	I-protein
,	NN	O	O
IL-2R	NN	O	B-protein
beta-chain	NN	O	I-protein
,	NN	O	O
HLA-DR	NN	O	B-protein
,	NN	O	O
CD11a	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CD11c	NN	O	B-protein
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
IFN-gamma	NN	O	B-protein
activity	NN	O	O
but	NN	O	O
not	NN	O	O
IL-1	NN	O	O
,	NN	O	O
IL-2	NN	O	O
,	NN	O	O
or	NN	O	O
IL-4	NN	O	O
activity	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
culture	NN	O	O
supernatants	NN	O	O
72	NN	O	O
h	NN	O	O
after	NN	O	O
R24	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
some	NN	O	O
donors	NN	O	O
,	NN	O	O
increased	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
activity	NN	O	O
also	NN	O	O
was	NN	O	O
detected	NN	O	O
after	NN	O	O
R24	NN	O	B-protein
treatment	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
R24	NN	O	B-protein
treatment	NN	O	O
resulted	NN	O	O
in	NN	O	O
translocation	NN	O	O
of	NN	O	O
c-rel	NN	O	B-protein
,	NN	O	O
but	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
p50	NN	O	I-protein
or	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
from	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
containing	NN	O	O
c-rel	NN	O	B-protein
and	NN	O	O
p50	NN	O	B-protein
.	NN	O	O

This	NN	O	O
treatment	NN	O	O
also	NN	O	O
caused	NN	O	O
increased	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
specific	NN	O	O
protein	NN	O	O
substrates	NN	O	O
.	NN	O	O

R24	NN	O	B-protein
-stimulated	NN	O	O
increases	NN	O	O
in	NN	O	O
proliferation	NN	O	O
,	NN	O	O
cytotoxicity	NN	O	O
,	NN	O	O
and	NN	O	O
cell	NN	O	O
surface	NN	O	O
protein	NN	O	O
expression	NN	O	O
could	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
and	NN	O	O
staurosporin	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
cyclophilin/calcineurin	NN	O	B-protein
and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
R24	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
,	NN	O	O
a	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
inhibitor	NN	O	O
,	NN	O	O
blocked	NN	O	O
the	NN	O	O
R24	NN	O	B-protein
-stimulated	NN	O	O
increase	NN	O	O
in	NN	O	O
proliferation	NN	O	O
but	NN	O	O
not	NN	O	O
cytotoxicity	NN	O	O
at	NN	O	O
concentrations	NN	O	O
consistent	NN	O	O
with	NN	O	O
specificity	NN	O	O
for	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
multiple	NN	O	O
biochemical	NN	O	O
pathways	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
R24	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Genes	NN	O	O
encoding	NN	O	O
general	NN	O	B-protein
initiation	NN	O	I-protein
factors	NN	O	I-protein
for	NN	O	O
RNA	NN	O	B-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
transcription	NN	O	O
are	NN	O	O
dispersed	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
genome	NN	O	I-DNA
.	NN	O	O

General	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
accurate	NN	O	O
initiation	NN	O	O
of	NN	O	O
transcription	NN	O	O
by	NN	O	O
RNA	NN	O	B-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
.	NN	O	O

Human	NN	O	B-DNA
cDNAs	NN	O	I-DNA
encoding	NN	O	O
subunits	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
have	NN	O	O
been	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
.	NN	O	O

Using	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
genes	NN	O	B-DNA
encoding	NN	O	O
the	NN	O	O
TATA-box	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
TBP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
TFIIB	NN	O	B-protein
,	NN	O	O
TFIIE	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
TFIIE	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
RAP30	NN	O	B-protein
,	NN	O	O
RAP74	NN	O	B-protein
and	NN	O	O
the	NN	O	O
62	NN	O	B-protein
kDa	NN	O	I-protein
subunit	NN	O	I-protein
,	NN	O	O
of	NN	O	O
TFIIH	NN	O	B-protein
are	NN	O	O
located	NN	O	O
at	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
chromosomal	NN	O	I-DNA
bands	NN	O	I-DNA
6q26-27	NN	O	I-DNA
,	NN	O	I-DNA
1p21-22	NN	O	I-DNA
,	NN	O	I-DNA
3q21-24	NN	O	I-DNA
,	NN	O	I-DNA
8p12	NN	O	I-DNA
,	NN	O	I-DNA
13q14	NN	O	I-DNA
,	NN	O	I-DNA
19p13.3	NN	O	I-DNA
and	NN	O	I-DNA
11p14-15.1	NN	O	I-DNA
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

This	NN	O	O
dispersed	NN	O	O
localization	NN	O	O
of	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
functionally	NN	O	O
related	NN	O	O
gene	NN	O	O
provides	NN	O	O
insights	NN	O	O
into	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
genome	NN	O	I-DNA
evolution	NN	O	O
and	NN	O	O
their	NN	O	O
possible	NN	O	O
involvement	NN	O	O
in	NN	O	O
human	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

BCL-6	NN	O	B-protein
and	NN	O	O
the	NN	O	O
molecular	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
B-cell	NN	O	O
lymphoma	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
presented	NN	O	O
identify	NN	O	O
the	NN	O	O
first	NN	O	O
genetic	NN	O	O
lesion	NN	O	O
associated	NN	O	O
with	NN	O	O
DLCL	NN	O	O
,	NN	O	O
the	NN	O	O
most	NN	O	O
clinically	NN	O	O
relevant	NN	O	O
form	NN	O	O
of	NN	O	O
NHL	NN	O	O
.	NN	O	O

Although	NN	O	O
no	NN	O	O
proof	NN	O	O
yet	NN	O	O
exists	NN	O	O
of	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
these	NN	O	O
lesions	NN	O	O
in	NN	O	O
DLCL	NN	O	O
pathogenesis	NN	O	O
,	NN	O	O
the	NN	O	O
feature	NN	O	O
of	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
,	NN	O	O
its	NN	O	O
specific	NN	O	O
pattern	NN	O	O
of	NN	O	O
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
clustering	NN	O	O
of	NN	O	O
lesions	NN	O	O
disrupting	NN	O	O
its	NN	O	O
regulatory	NN	O	O
domain	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
deregulation	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
expression	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
DLCL	NN	O	O
development	NN	O	O
.	NN	O	O

A	NN	O	O
more	NN	O	O
precise	NN	O	O
definition	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
in	NN	O	O
normal	NN	O	O
and	NN	O	O
neoplastic	NN	O	O
B-cell	NN	O	O
development	NN	O	O
is	NN	O	O
the	NN	O	O
goal	NN	O	O
of	NN	O	O
ongoing	NN	O	O
study	NN	O	O
of	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
engineered	NN	O	O
either	NN	O	O
to	NN	O	O
express	NN	O	O
BCL-6	NN	O	B-protein
under	NN	O	O
heterologous	NN	O	B-DNA
promoters	NN	O	I-DNA
or	NN	O	O
lacking	NN	O	O
BCL-6	NN	O	B-protein
function	NN	O	O
due	NN	O	O
to	NN	O	O
targeted	NN	O	O
deletions	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
contributing	NN	O	O
to	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
DLCL	NN	O	O
pathogenesis	NN	O	O
,	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
lesions	NN	O	O
may	NN	O	O
have	NN	O	O
relevant	NN	O	O
clinical	NN	O	O
implications	NN	O	O
.	NN	O	O

DLCL	NN	O	O
represent	NN	O	O
a	NN	O	O
heterogeneous	NN	O	O
group	NN	O	O
of	NN	O	O
neoplasms	NN	O	O
which	NN	O	O
are	NN	O	O
treated	NN	O	O
homogeneously	NN	O	O
despite	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
only	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
experience	NN	O	O
long-term	NN	O	O
disease-free	NN	O	O
survival	NN	O	O
(	NN	O	O
Schneider	NN	O	O
et	NN	O	O
al.	NN	O	O
1990	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
fact	NN	O	O
that	NN	O	O
BCL-6	NN	O	B-protein
rearrangements	NN	O	O
identify	NN	O	O
biologically	NN	O	O
and	NN	O	O
clinically	NN	O	O
distinct	NN	O	O
subsets	NN	O	O
of	NN	O	O
DLCL	NN	O	O
suggests	NN	O	O
that	NN	O	O
these	NN	O	O
lesions	NN	O	O
may	NN	O	O
be	NN	O	O
useful	NN	O	O
as	NN	O	O
markers	NN	O	O
in	NN	O	O
selection	NN	O	O
of	NN	O	O
differential	NN	O	O
therapeutic	NN	O	O
strategies	NN	O	O
based	NN	O	O
on	NN	O	O
different	NN	O	O
risk	NN	O	O
groups	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
rearrangements	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
and	NN	O	O
monitor	NN	O	O
the	NN	O	O
malignant	NN	O	B-cell_type
clone	NN	O	I-cell_type
with	NN	O	O
sensitive	NN	O	O
PCR-based	NN	O	O
techniques	NN	O	O
.	NN	O	O

Since	NN	O	O
clinical	NN	O	O
remission	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
a	NN	O	O
significant	NN	O	O
fraction	NN	O	O
of	NN	O	O
DLCL	NN	O	O
cases	NN	O	O
,	NN	O	O
these	NN	O	O
markers	NN	O	O
may	NN	O	O
serve	NN	O	O
as	NN	O	O
critical	NN	O	O
tools	NN	O	O
for	NN	O	O
sensitive	NN	O	O
monitoring	NN	O	O
of	NN	O	O
minimal	NN	O	O
residual	NN	O	O
disease	NN	O	O
and	NN	O	O
early	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
relapse	NN	O	O
(	NN	O	O
Gribben	NN	O	O
et	NN	O	O
al.	NN	O	O
1993	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Pancreatic	NN	O	O
development	NN	O	O
and	NN	O	O
maturation	NN	O	O
of	NN	O	O
the	NN	O	O
islet	NN	O	B-cell_type
B	NN	O	I-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
pluripotent	NN	O	B-cell_line
islet	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

Pancreas	NN	O	O
organogenesis	NN	O	O
is	NN	O	O
a	NN	O	O
highly	NN	O	O
regulated	NN	O	O
process	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
two	NN	O	O
anlage	NN	O	O
evaginate	NN	O	O
from	NN	O	O
the	NN	O	O
primitive	NN	O	O
gut	NN	O	O
.	NN	O	O

They	NN	O	O
later	NN	O	O
fuse	NN	O	O
,	NN	O	O
and	NN	O	O
,	NN	O	O
under	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
the	NN	O	O
surrounding	NN	O	O
mesenchyme	NN	O	O
,	NN	O	O
the	NN	O	O
mature	NN	O	O
organ	NN	O	O
develops	NN	O	O
,	NN	O	O
being	NN	O	O
mainly	NN	O	O
composed	NN	O	O
of	NN	O	O
ductal	NN	O	O
,	NN	O	O
exocrine	NN	O	O
and	NN	O	O
endocrine	NN	O	O
compartments	NN	O	O
.	NN	O	O

Early	NN	O	O
buds	NN	O	O
are	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
branching	NN	O	O
morphogenesis	NN	O	O
of	NN	O	O
the	NN	O	O
ductal	NN	O	O
epithelium	NN	O	O
from	NN	O	O
which	NN	O	O
endocrine	NN	O	B-cell_type
and	NN	O	I-cell_type
exocrine	NN	O	I-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
bud	NN	O	O
to	NN	O	O
eventually	NN	O	O
form	NN	O	O
the	NN	O	O
two	NN	O	O
other	NN	O	O
compartments	NN	O	O
.	NN	O	O

The	NN	O	O
three	NN	O	O
compartments	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
of	NN	O	O
common	NN	O	O
endodermal	NN	O	O
origin	NN	O	O
;	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
earlier	NN	O	O
hypotheses	NN	O	O
,	NN	O	O
which	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
endocrine	NN	O	O
compartment	NN	O	O
was	NN	O	O
of	NN	O	O
neuroectodermal	NN	O	O
origin	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
thus	NN	O	O
generally	NN	O	O
believed	NN	O	O
that	NN	O	O
the	NN	O	O
pancreatic	NN	O	O
endocrine-lineage	NN	O	O
possesses	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
mature	NN	O	O
along	NN	O	O
a	NN	O	O
differentiation	NN	O	O
pathway	NN	O	O
that	NN	O	O
shares	NN	O	O
many	NN	O	O
characteristics	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
neuronal	NN	O	O
differentiation	NN	O	O
.	NN	O	O

During	NN	O	O
recent	NN	O	O
years	NN	O	O
,	NN	O	O
studies	NN	O	O
of	NN	O	O
insulin-gene	NN	O	B-DNA
regulation	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
insulin-gene	NN	O	B-DNA
activity	NN	O	O
have	NN	O	O
provided	NN	O	O
information	NN	O	O
on	NN	O	O
pancreas	NN	O	O
development	NN	O	O
in	NN	O	O
general	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
review	NN	O	O
summarizes	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
special	NN	O	O
focus	NN	O	O
on	NN	O	O
our	NN	O	O
own	NN	O	O
studies	NN	O	O
on	NN	O	O
pluripotent	NN	O	B-cell_line
endocrine	NN	O	I-cell_line
cultures	NN	O	I-cell_line
of	NN	O	O
rat	NN	O	O
pancreas	NN	O	O
.	NN	O	O

-DOCSTART-	O

Octamer	NN	O	O
independent	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
germline	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Previous	NN	O	O
analyses	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
V	NN	O	I-DNA
region	NN	O	I-DNA
promoters	NN	O	I-DNA
has	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
discovery	NN	O	O
of	NN	O	O
a	NN	O	O
common	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
which	NN	O	O
is	NN	O	O
functionally	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
and	NN	O	O
developmentally	NN	O	O
regulated	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
germline	NN	O	B-DNA
promoters	NN	O	I-DNA
(	NN	O	O
Ko	NN	O	B-DNA
)	NN	O	O
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
J	NN	O	B-DNA
region	NN	O	I-DNA
gene	NN	O	I-DNA
segments	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
locus	NN	O	I-DNA
also	NN	O	O
contain	NN	O	O
an	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
(	NN	O	O
containing	NN	O	O
a	NN	O	O
single	NN	O	O
base	NN	O	B-DNA
pair	NN	O	I-DNA
mutation	NN	O	I-DNA
and	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
the	NN	O	O
variant	NN	O	O
octamer	NN	O	O
)	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
previously	NN	O	O
to	NN	O	O
bind	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
elucidate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
variant	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
germline	NN	O	B-DNA
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
unrearranged	NN	O	O
kappa	NN	O	B-DNA
locus	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
have	NN	O	O
quantitated	NN	O	O
the	NN	O	O
relative	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
for	NN	O	O
the	NN	O	O
variant	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
and	NN	O	O
determined	NN	O	O
the	NN	O	O
functional	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
in	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
variant	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
binds	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
5-fold	NN	O	O
lower	NN	O	O
affinity	NN	O	O
than	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
octamer	NN	O	I-DNA
motif	NN	O	I-DNA
,	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
variant	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
to	NN	O	O
either	NN	O	O
a	NN	O	O
consensus	NN	O	B-DNA
octamer	NN	O	I-DNA
or	NN	O	O
non-octamer	NN	O	B-DNA
motif	NN	O	I-DNA
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
from	NN	O	O
the	NN	O	O
germline	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
also	NN	O	O
find	NN	O	O
significant	NN	O	O
differences	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
germline	NN	O	B-DNA
and	NN	O	O
V	NN	O	B-DNA
region	NN	O	I-DNA
promoters	NN	O	I-DNA
by	NN	O	O
kappa	NN	O	B-DNA
enhancers	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
germline	NN	O	B-DNA
promoters	NN	O	I-DNA
and	NN	O	O
V	NN	O	B-DNA
region	NN	O	I-DNA
promoters	NN	O	I-DNA
differ	NN	O	O
in	NN	O	O
their	NN	O	O
dependence	NN	O	O
on	NN	O	O
octamer	NN	O	B-DNA
for	NN	O	O
activation	NN	O	O
and	NN	O	O
respond	NN	O	O
differently	NN	O	O
to	NN	O	O
enhancer	NN	O	O
activation	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
have	NN	O	O
important	NN	O	O
implications	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
germline	NN	O	B-DNA
transcription	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
concomitant	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
V-J	NN	O	O
recombination	NN	O	O
of	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
locus	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Generation	NN	O	O
of	NN	O	O
CD1+RelB+	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
tartrate-resistant	NN	O	B-cell_type
acid	NN	O	I-cell_type
phosphatase-positive	NN	O	I-cell_type
osteoclast-like	NN	O	I-cell_type
multinucleated	NN	O	I-cell_type
giant	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
previously	NN	O	O
showed	NN	O	O
that	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
M-CSF	NN	O	B-protein
)	NN	O	O
stimulate	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
into	NN	O	O
two	NN	O	O
phenotypically	NN	O	O
distinct	NN	O	O
types	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
not	NN	O	O
only	NN	O	O
CSF	NN	O	B-protein
but	NN	O	O
also	NN	O	O
many	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
are	NN	O	O
produced	NN	O	O
under	NN	O	O
various	NN	O	O
conditions	NN	O	O
.	NN	O	O

Those	NN	O	O
cytokines	NN	O	B-protein
may	NN	O	O
modulate	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
by	NN	O	O
CSFs	NN	O	B-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
CD14+	NN	O	B-cell_type
adherent	NN	O	I-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
can	NN	O	O
differentiate	NN	O	O
into	NN	O	O
CD1+relB+	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
DC	NN	O	B-cell_type
)	NN	O	O
by	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
plus	NN	O	O
interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
and	NN	O	O
that	NN	O	O
they	NN	O	O
differentiate	NN	O	O
into	NN	O	O
tartrate-resistant	NN	O	B-cell_type
acid	NN	O	I-cell_type
phosphatase	NN	O	I-cell_type
(	NN	O	I-cell_type
TRAP	NN	O	I-cell_type
)	NN	O	I-cell_type
-positive	NN	O	I-cell_type
osteoclast-like	NN	O	I-cell_type
multinucleated	NN	O	I-cell_type
giant	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
MGC	NN	O	B-cell_type
)	NN	O	O
by	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
M-CSF	NN	O	B-protein
plus	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
monocyte-derived	NN	O	B-cell_line
DC	NN	O	I-cell_line
were	NN	O	O
not	NN	O	O
terminally	NN	O	B-cell_line
differentiated	NN	O	I-cell_line
cells	NN	O	I-cell_line
;	NN	O	O
they	NN	O	O
could	NN	O	O
still	NN	O	O
convert	NN	O	O
to	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
M-CSF	NN	O	B-protein
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
stimulated	NN	O	O
the	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
DC	NN	O	B-cell_type
by	NN	O	O
downregulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
cfms	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
aborting	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
convert	NN	O	O
to	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
had	NN	O	O
no	NN	O	O
demonstrable	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
IFN-gamma	NN	O	B-protein
antagonized	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
suppressed	NN	O	O
the	NN	O	O
DC	NN	O	B-cell_type
and	NN	O	O
MGC	NN	O	B-cell_type
formation	NN	O	O
induced	NN	O	O
by	NN	O	O
GM-CSF	NN	O	B-protein
+	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
M-CSF	NN	O	B-protein
+	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
provide	NN	O	O
a	NN	O	O
new	NN	O	O
aspect	NN	O	O
to	NN	O	O
our	NN	O	O
knowledge	NN	O	O
of	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
and	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
are	NN	O	O
flexible	NN	O	O
in	NN	O	O
their	NN	O	O
differentiation	NN	O	O
potential	NN	O	O
and	NN	O	O
are	NN	O	O
precursors	NN	O	O
not	NN	O	O
only	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
but	NN	O	O
also	NN	O	O
of	NN	O	O
CD1+relB+DC	NN	O	B-cell_type
and	NN	O	O
TRAP-positive	NN	O	B-cell_type
MGC	NN	O	I-cell_type
.	NN	O	O

Such	NN	O	O
a	NN	O	O
diverse	NN	O	O
pathway	NN	O	O
of	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
may	NN	O	O
constitute	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
basic	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
immune	NN	O	O
regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibitor	NN	O	O
,	NN	O	O
tepoxalin	NN	O	O
,	NN	O	O
suppresses	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	B-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
CD62E	NN	O	B-protein
,	NN	O	O
CD11b/CD18	NN	O	B-protein
and	NN	O	O
CD106	NN	O	B-protein
.	NN	O	O

Tepoxalin	NN	O	O
,	NN	O	O
a	NN	O	O
dual	NN	O	O
enzyme	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
cyclooxygenase	NN	O	B-protein
and	NN	O	O
5-lipoxygenase	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
inhibit	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Its	NN	O	O
immunosuppressive	NN	O	O
property	NN	O	O
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
cyclosporin	NN	O	O
because	NN	O	O
only	NN	O	O
tepoxalin	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
cyclosporin	NN	O	O
,	NN	O	O
suppresses	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
tepoxalin	NN	O	O
selectively	NN	O	O
inhibits	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	B-protein
ICAM-1	NN	O	I-protein
,	NN	O	I-protein
CD54	NN	O	I-protein
)	NN	O	I-protein
/MAC-1	NN	O	I-protein
(	NN	O	O
CD11b/CD18	NN	O	B-protein
)	NN	O	O
dependent	NN	O	O
adhesion	NN	O	O
of	NN	O	O
polymorphonuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
IL-1	NN	O	B-protein
activated	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
inhibition	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
cell	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

Flow	NN	O	O
cytometry	NN	O	O
analyses	NN	O	O
on	NN	O	O
cultured	NN	O	O
cells	NN	O	O
that	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
tepoxalin	NN	O	O
or	NN	O	O
antisense	NN	O	O
oligonucleotides	NN	O	O
to	NN	O	O
the	NN	O	O
P65/p50	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
then	NN	O	O
stimulated	NN	O	O
with	NN	O	O
PMA	NN	O	O
,	NN	O	O
revealed	NN	O	O
a	NN	O	O
reduced	NN	O	O
expression	NN	O	O
of	NN	O	O
CD11b/CD18	NN	O	B-protein
on	NN	O	O
monocytic	NN	O	B-cell_line
HL60	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
endothelial	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
CD62E	NN	O	B-protein
)	NN	O	O
and	NN	O	O
vascular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
CD106	NN	O	B-protein
)	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
other	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
such	NN	O	O
as	NN	O	O
lymphocyte	NN	O	B-protein
function	NN	O	I-protein
associated-antigen-1	NN	O	I-protein
(	NN	O	O
CD11a/CD18	NN	O	B-protein
)	NN	O	O
and	NN	O	O
CD54	NN	O	B-protein
were	NN	O	O
unaffected	NN	O	O
.	NN	O	O

Tepoxalin	NN	O	O
also	NN	O	O
inhibited	NN	O	O
the	NN	O	O
secretion	NN	O	O
of	NN	O	O
a	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
regulated	NN	O	I-protein
chemokine	NN	O	I-protein
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
a	NN	O	O
known	NN	O	O
inducer	NN	O	O
of	NN	O	O
CD11b/CD18	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Thus	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
CD11b/CD18	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
tepoxalin	NN	O	O
may	NN	O	O
involve	NN	O	O
IL-8	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
can	NN	O	O
be	NN	O	O
modulated	NN	O	O
and	NN	O	O
that	NN	O	O
tepoxalin	NN	O	O
may	NN	O	O
be	NN	O	O
useful	NN	O	O
in	NN	O	O
treating	NN	O	O
selected	NN	O	O
adhesion	NN	O	O
mediated	NN	O	O
events	NN	O	O
such	NN	O	O
as	NN	O	O
leukocyte	NN	O	O
migration	NN	O	O
or	NN	O	O
atherosclerotic	NN	O	O
plaque	NN	O	O
formation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
a	NN	O	O
CD43/leukosialin	NN	O	B-protein
-mediated	NN	O	O
pathway	NN	O	O
for	NN	O	O
inducing	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T-lymphoblastoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
(	NN	O	O
mAb	NN	O	B-protein
)	NN	O	O
J393	NN	O	B-protein
induces	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
.	NN	O	O

NH2	NN	O	B-protein
-terminal	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
identified	NN	O	O
the	NN	O	O
140-kDa	NN	O	B-protein
surface	NN	O	I-protein
antigen	NN	O	I-protein
for	NN	O	O
mAb	NN	O	B-protein
J393	NN	O	I-protein
as	NN	O	O
CD43/leukosialin	NN	O	B-protein
,	NN	O	O
the	NN	O	O
major	NN	O	O
sialoglycoprotein	NN	O	B-protein
of	NN	O	O
leukocytes	NN	O	B-cell_type
.	NN	O	O

While	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
co-expressed	NN	O	O
two	NN	O	O
discrete	NN	O	O
cell-surface	NN	O	B-protein
isoforms	NN	O	I-protein
of	NN	O	O
CD43	NN	O	B-protein
,	NN	O	O
recognized	NN	O	O
by	NN	O	O
mAb	NN	O	B-protein
J393	NN	O	I-protein
and	NN	O	O
mAb	NN	O	B-protein
G10-2	NN	O	I-protein
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
only	NN	O	O
J393/CD43	NN	O	B-protein
signaled	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

J393/CD43	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
hyposialylated	NN	O	O
,	NN	O	O
bearing	NN	O	O
predominantly	NN	O	O
O-linked	NN	O	O
monosaccharide	NN	O	O
glycans	NN	O	O
,	NN	O	O
whereas	NN	O	O
G10-2/CD43	NN	O	B-protein
bore	NN	O	O
complex	NN	O	O
sialylated	NN	O	O
tetra-	NN	O	O
and	NN	O	O
hexasaccharide	NN	O	O
chains	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
soluble	NN	O	O
,	NN	O	O
bivalent	NN	O	O
mAb	NN	O	B-protein
J393	NN	O	I-protein
killed	NN	O	O
25-50	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	B-cell_line
population	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
concomitant	NN	O	O
engagement	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
CD3.TcR	NN	O	B-protein
complex	NN	O	I-protein
or	NN	O	O
the	NN	O	O
integrins	NN	O	B-protein
CD18	NN	O	B-protein
and	NN	O	O
CD29	NN	O	B-protein
significantly	NN	O	O
potentiated	NN	O	O
this	NN	O	O
effect	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
mAb	NN	O	B-protein
J393	NN	O	I-protein
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
specific	NN	O	O
protein	NN	O	B-protein
substrates	NN	O	I-protein
that	NN	O	O
underwent	NN	O	O
hyperphosphorylation	NN	O	O
upon	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
costimulation	NN	O	O
.	NN	O	O

Tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
inhibition	NN	O	O
by	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
diminished	NN	O	O
J393/CD43	NN	O	B-protein
-mediated	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
whereas	NN	O	O
inhibition	NN	O	O
of	NN	O	O
phosphotyrosine	NN	O	B-protein
phosphatase	NN	O	I-protein
activity	NN	O	O
by	NN	O	O
bis	NN	O	O
(	NN	O	O
maltolato	NN	O	O
)	NN	O	O
oxovanadium-IV	NN	O	O
enhanced	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
activation	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
altered	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
as	NN	O	O
J393/	NN	O	O
CD43	NN	O	B-protein
ligation	NN	O	O
prompted	NN	O	O
decreases	NN	O	O
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
protein	NN	O	I-protein
NF-kappaB	NN	O	B-protein
and	NN	O	O
proteins	NN	O	O
binding	NN	O	O
the	NN	O	O
interferon-inducible	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Since	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
express	NN	O	O
cryptic	NN	O	B-protein
epitopes	NN	O	I-protein
for	NN	O	O
mAb	NN	O	B-protein
J393	NN	O	I-protein
,	NN	O	O
these	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
tightly	NN	O	O
regulated	NN	O	O
CD43	NN	O	B-protein
-mediated	NN	O	O
pathway	NN	O	O
for	NN	O	O
inducing	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Sterol	NN	O	O
dependent	NN	O	O
LDL-receptor	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
normal	NN	O	O
and	NN	O	O
CML	NN	O	O
patients	NN	O	O
.	NN	O	O

Sterol	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
SRE	NN	O	B-DNA
)	NN	O	O
has	NN	O	O
been	NN	O	O
recognized	NN	O	O
to	NN	O	O
regulate	NN	O	O
various	NN	O	O
key	NN	O	O
genes	NN	O	B-DNA
coding	NN	O	O
for	NN	O	O
especially	NN	O	O
low	NN	O	B-protein
density	NN	O	I-protein
lipoprotein	NN	O	I-protein
(	NN	O	I-protein
LDL	NN	O	I-protein
)	NN	O	I-protein
-receptor	NN	O	I-protein
,	NN	O	O
3-hydroxy-3-methylglutaryl	NN	O	B-protein
coenzyme	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	I-protein
HMG-CoA	NN	O	I-protein
)	NN	O	I-protein
reductase	NN	O	I-protein
and	NN	O	O
HMG-CoA	NN	O	B-protein
synthase	NN	O	I-protein
known	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
cholesterol	NN	O	O
feedback	NN	O	O
mechanism	NN	O	O
.	NN	O	O

The	NN	O	O
deranged	NN	O	O
cholesterol	NN	O	O
feedback	NN	O	O
mechanism	NN	O	O
has	NN	O	O
been	NN	O	O
widely	NN	O	O
recognised	NN	O	O
in	NN	O	O
initiation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
progression	NN	O	O
of	NN	O	O
various	NN	O	O
types	NN	O	O
of	NN	O	O
cancers	NN	O	O
including	NN	O	O
chronic	NN	O	O
myeloid	NN	O	O
leukaemia	NN	O	O
(	NN	O	O
CML	NN	O	O
)	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
addressed	NN	O	O
to	NN	O	O
understand	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
and	NN	O	O
revealed	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
unique	NN	O	B-protein
47	NN	O	I-protein
kDa	NN	O	I-protein
protein	NN	O	I-protein
factor	NN	O	I-protein
having	NN	O	O
affinity	NN	O	O
for	NN	O	O
this	NN	O	O
SRE	NN	O	B-DNA
sequence	NN	O	I-DNA
in	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
normal	NN	O	O
subjects	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
its	NN	O	O
absence	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
untreated	NN	O	O
CML	NN	O	O
patients	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
factor	NN	O	O
appeared	NN	O	O
when	NN	O	O
the	NN	O	O
CML	NN	O	O
patients	NN	O	O
achieved	NN	O	O
complete	NN	O	O
haematological	NN	O	O
remission	NN	O	O
(	NN	O	O
CHR	NN	O	O
)	NN	O	O
through	NN	O	O
alpha-interferon	NN	O	B-protein
therapy	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
an	NN	O	O
inverse	NN	O	O
relationship	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
between	NN	O	O
sterol	NN	O	O
modulated	NN	O	O
LDL-receptor	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
and	NN	O	O
the	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
this	NN	O	O
47	NN	O	B-protein
kDa	NN	O	I-protein
factor	NN	O	I-protein
to	NN	O	O
the	NN	O	O
SRE	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Based	NN	O	O
upon	NN	O	O
these	NN	O	O
results	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
alpha-interferon	NN	O	B-protein
through	NN	O	O
its	NN	O	O
receptor	NN	O	O
initiates	NN	O	O
phosphatidic	NN	O	O
acid	NN	O	O
dependent	NN	O	O
signalling	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
regulates	NN	O	O
the	NN	O	O
affinity	NN	O	O
of	NN	O	O
47	NN	O	B-protein
kDa	NN	O	I-protein
sterol	NN	O	I-protein
regulatory	NN	O	I-protein
element	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
LDL-receptor	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
CML	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Multiple	NN	O	O
prolactin-responsive	NN	O	B-DNA
elements	NN	O	I-DNA
mediate	NN	O	O
G1	NN	O	O
and	NN	O	O
S	NN	O	O
phase	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
interferon	NN	O	B-DNA
regulatory	NN	O	I-DNA
factor-1	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
interferon	NN	O	B-DNA
regulatory	NN	O	I-DNA
factor-1	NN	O	I-DNA
(	NN	O	I-DNA
IRF-1	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
both	NN	O	O
an	NN	O	O
immediate-early	NN	O	B-DNA
G1	NN	O	I-DNA
phase	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
an	NN	O	O
S	NN	O	B-DNA
phase	NN	O	I-DNA
gene	NN	O	I-DNA
inducible	NN	O	O
by	NN	O	O
PRL	NN	O	B-protein
in	NN	O	O
rat	NN	O	B-cell_line
Nb2	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
PRL	NN	O	B-protein
regulates	NN	O	O
the	NN	O	O
biphasic	NN	O	O
expression	NN	O	O
of	NN	O	O
IRF-1	NN	O	B-protein
,	NN	O	O
we	NN	O	O
cloned	NN	O	O
the	NN	O	O
rat	NN	O	B-DNA
IRF-1	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
functionally	NN	O	O
characterized	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Upon	NN	O	O
transfection	NN	O	O
into	NN	O	O
Nb2	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
1.7	NN	O	O
kilobases	NN	O	O
(	NN	O	O
kb	NN	O	O
)	NN	O	O
of	NN	O	O
IRF-1	NN	O	B-DNA
5'-flanking	NN	O	I-DNA
DNA	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
a	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyl	NN	O	I-DNA
transferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
mediated	NN	O	O
a	NN	O	O
30-fold	NN	O	O
induction	NN	O	O
of	NN	O	O
CAT	NN	O	B-protein
enzyme	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
24	NN	O	O
h	NN	O	O
of	NN	O	O
PRL	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Deletion	NN	O	O
mutants	NN	O	O
containing	NN	O	O
1.3	NN	O	B-DNA
,	NN	O	I-DNA
0.6	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
0.2	NN	O	I-DNA
kb	NN	O	I-DNA
5'-flanking	NN	O	I-DNA
DNA	NN	O	I-DNA
were	NN	O	O
incrementally	NN	O	O
less	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
,	NN	O	O
although	NN	O	O
0.2	NN	O	O
kb	NN	O	O
still	NN	O	O
mediated	NN	O	O
a	NN	O	O
12-fold	NN	O	O
induction	NN	O	O
by	NN	O	O
PRL	NN	O	B-protein
.	NN	O	O

The	NN	O	O
sequence	NN	O	O
between	NN	O	O
-1.7	NN	O	O
and	NN	O	O
-0.2	NN	O	O
kb	NN	O	O
linked	NN	O	O
to	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
thymidine	NN	O	I-DNA
kinase	NN	O	I-DNA
promoter	NN	O	I-DNA
failed	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
PRL	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
upstream	NN	O	B-DNA
PRL	NN	O	I-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
may	NN	O	O
require	NN	O	O
an	NN	O	O
interaction	NN	O	O
with	NN	O	O
promoter-proximal	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

By	NN	O	O
assaying	NN	O	O
CAT	NN	O	B-protein
enzyme	NN	O	I-protein
activity	NN	O	O
across	NN	O	O
a	NN	O	O
24-h	NN	O	O
PRL	NN	O	B-protein
induction	NN	O	O
time	NN	O	O
course	NN	O	O
,	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
assign	NN	O	O
G1	NN	O	O
vs.	NN	O	O
S	NN	O	O
phase	NN	O	O
PRL	NN	O	B-protein
responses	NN	O	O
of	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-DNA
gene	NN	O	I-DNA
to	NN	O	O
different	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-protein
5'-flanking	NN	O	B-DNA
and	NN	O	O
promoter	NN	O	B-DNA
DNA	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
0.2-kb	NN	O	O
IRF-CAT	NN	O	B-DNA
construct	NN	O	I-DNA
was	NN	O	O
induced	NN	O	O
by	NN	O	O
PRL	NN	O	B-protein
stimulation	NN	O	O
during	NN	O	O
the	NN	O	O
G1	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
1.7-kb	NN	O	O
IRF-	NN	O	O
CAT	NN	O	B-protein
construct	NN	O	O
was	NN	O	O
inducible	NN	O	O
by	NN	O	O
PRL	NN	O	B-protein
during	NN	O	O
both	NN	O	O
G1	NN	O	O
and	NN	O	O
S	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
the	NN	O	O
PRL	NN	O	B-protein
-induced	NN	O	O
biphasic	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-DNA
gene	NN	O	I-DNA
appears	NN	O	O
to	NN	O	O
be	NN	O	O
controlled	NN	O	O
by	NN	O	O
separate	NN	O	O
PRL-responsive	NN	O	B-DNA
elements	NN	O	I-DNA
:	NN	O	O
elements	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	O
0.2	NN	O	O
kb	NN	O	O
of	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
act	NN	O	O
during	NN	O	O
early	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
elements	NN	O	O
between	NN	O	O
0.2	NN	O	O
and	NN	O	O
1.7	NN	O	O
kb	NN	O	O
act	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
the	NN	O	O
proximal	NN	O	O
0.2-kb	NN	O	O
region	NN	O	O
during	NN	O	O
S	NN	O	O
phase	NN	O	O
progression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
BZLF1	NN	O	B-DNA
promoter	NN	O	I-DNA
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
by	NN	O	O
second	NN	O	O
messengers	NN	O	O
in	NN	O	O
anti-immunoglobulin-treated	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Initiation	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
BZLF1	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
BZLF1	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
(	NN	O	O
Zp	NN	O	B-DNA
)	NN	O	O
was	NN	O	O
activated	NN	O	O
by	NN	O	O
crosslinking	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
immunoglobulin	NN	O	I-protein
(	NN	O	O
Ig	NN	O	B-protein
)	NN	O	O
with	NN	O	O
anti-Ig	NN	O	B-protein
antibody	NN	O	I-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
other	NN	O	O
viral	NN	O	O
genes	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
several	NN	O	O
anti-Ig	NN	O	O
response	NN	O	O
elements	NN	O	O
within	NN	O	O
Zp	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
were	NN	O	O
originally	NN	O	O
defined	NN	O	O
as	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	B-DNA
(	NN	O	I-DNA
TPA	NN	O	I-DNA
)	NN	O	I-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
(	NN	O	O
ZI	NN	O	O
repeats	NN	O	O
and	NN	O	O
ZII	NN	O	B-DNA
,	NN	O	O
an	NN	O	O
AP-1-like	NN	O	B-DNA
domain	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

Since	NN	O	O
anti-Ig	NN	O	O
crosslinking	NN	O	O
leads	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
level	NN	O	O
,	NN	O	O
Zp	NN	O	B-DNA
was	NN	O	O
tested	NN	O	O
for	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
these	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
A23187	NN	O	O
increased	NN	O	O
Zp	NN	O	B-DNA
activity	NN	O	O
.	NN	O	O

When	NN	O	O
the	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
was	NN	O	O
used	NN	O	O
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
TPA	NN	O	O
,	NN	O	O
a	NN	O	O
PKC	NN	O	B-protein
activator	NN	O	O
,	NN	O	O
the	NN	O	O
Zp	NN	O	B-DNA
induction	NN	O	O
was	NN	O	O
synergistically	NN	O	O
enhanced	NN	O	O
.	NN	O	O

1-	NN	O	O
(	NN	O	O
5-Isoquinolinyl	NN	O	O
sulfonyl	NN	O	O
)	NN	O	O
-2-methylpiperazine	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
,	NN	O	O
inhibited	NN	O	O
the	NN	O	O
anti-Ig	NN	O	B-protein
inducibility	NN	O	O
of	NN	O	O
Zp	NN	O	B-DNA
.	NN	O	O

Calmodulin	NN	O	O
antagonists	NN	O	O
,	NN	O	O
compound	NN	O	O
R24571	NN	O	O
and	NN	O	O
trifluoperazine	NN	O	O
,	NN	O	O
blocked	NN	O	O
the	NN	O	O
Zp	NN	O	B-DNA
activation	NN	O	O
with	NN	O	O
anti-Ig	NN	O	B-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
Zp	NN	O	B-DNA
responds	NN	O	O
directly	NN	O	O
to	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
both	NN	O	O
PKC	NN	O	B-protein
and	NN	O	O
calcium/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

Requirement	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activation	NN	O	O
for	NN	O	O
the	NN	O	O
anti-Ig	NN	O	B-protein
-mediated	NN	O	O
Zp	NN	O	B-DNA
activation	NN	O	O
was	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
through	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
herbimycin	NN	O	O
.	NN	O	O

These	NN	O	O
cellular	NN	O	B-protein
gene	NN	O	I-protein
regulatory	NN	O	I-protein
molecules	NN	O	I-protein
induced	NN	O	O
with	NN	O	O
anti-Ig	NN	O	O
may	NN	O	O
cooperatively	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
part	NN	O	O
in	NN	O	O
achieving	NN	O	O
efficient	NN	O	O
EBV	NN	O	O
activation	NN	O	O
as	NN	O	O
seen	NN	O	O
with	NN	O	O
anti-Ig	NN	O	B-protein
treatment	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
SCL	NN	O	B-protein
protein	NN	O	I-protein
displays	NN	O	O
cell-specific	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
size	NN	O	O
.	NN	O	O

SCL	NN	O	B-protein
protein	NN	O	I-protein
production	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
hemopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
by	NN	O	O
immunoblotting	NN	O	O
using	NN	O	O
specific	NN	O	O
polyclonal	NN	O	O
antisera	NN	O	O
.	NN	O	O

SCL	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_line
,	NN	O	I-cell_line
megakaryocyte	NN	O	I-cell_line
,	NN	O	I-cell_line
mast	NN	O	I-cell_line
and	NN	O	I-cell_line
early	NN	O	I-cell_line
myeloid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
several	NN	O	O
lymphoid	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
which	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
harbor	NN	O	O
SCL	NN	O	B-DNA
gene	NN	O	I-DNA
rearrangements	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
most	NN	O	O
cell	NN	O	O
lines	NN	O	O
,	NN	O	O
proteins	NN	O	O
of	NN	O	O
molecular	NN	O	O
weight	NN	O	O
49	NN	O	O
and	NN	O	O
44	NN	O	O
kDa	NN	O	O
were	NN	O	O
found	NN	O	O
,	NN	O	O
however	NN	O	O
two	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
lines	NN	O	O
expressed	NN	O	O
only	NN	O	O
lower	NN	O	O
molecular	NN	O	B-protein
weight	NN	O	I-protein
species	NN	O	I-protein
of	NN	O	I-protein
24	NN	O	I-protein
and	NN	O	I-protein
22	NN	O	I-protein
kDa	NN	O	I-protein
.	NN	O	O

This	NN	O	O
size	NN	O	O
discrepancy	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
cell-specific	NN	O	O
translational	NN	O	O
regulation	NN	O	O
,	NN	O	O
since	NN	O	O
overexpression	NN	O	O
of	NN	O	O
a	NN	O	O
retrovirally	NN	O	O
transfected	NN	O	O
SCL	NN	O	B-DNA
gene	NN	O	I-DNA
yielded	NN	O	O
the	NN	O	O
higher	NN	O	O
molecular	NN	O	O
weight	NN	O	O
forms	NN	O	O
in	NN	O	O
most	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
GP+E-86	NN	O	B-cell_line
,	NN	O	O
AT2.5	NN	O	B-cell_line
,	NN	O	O
M1	NN	O	B-cell_line
)	NN	O	O
but	NN	O	O
only	NN	O	O
the	NN	O	O
22	NN	O	B-protein
kDa	NN	O	I-protein
form	NN	O	I-protein
in	NN	O	O
the	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
WEHI-3B/D+	NN	O	B-cell_line
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
full-length	NN	O	O
SCL	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
the	NN	O	O
lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
SupT1	NN	O	O
and	NN	O	O
Raji	NN	O	O
,	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
cell	NN	O	O
phenotype	NN	O	O
and	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
evidence	NN	O	O
for	NN	O	O
autoregulation	NN	O	O
of	NN	O	O
SCL	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
restricted	NN	O	O
pattern	NN	O	O
of	NN	O	O
SCL	NN	O	B-protein
protein	NN	O	I-protein
synthesis	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
restricted	NN	O	O
expression	NN	O	O
of	NN	O	O
SCL	NN	O	B-RNA
mRNA	NN	O	I-RNA
documented	NN	O	O
previously	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
present	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
SCL	NN	O	B-protein
protein	NN	O	I-protein
size	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
regulation	NN	O	O
of	NN	O	O
translation	NN	O	O
in	NN	O	O
a	NN	O	O
cell-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

-DOCSTART-	O

Description	NN	O	O
and	NN	O	O
functional	NN	O	O
implications	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
sex-determining	NN	O	B-DNA
gene	NN	O	I-DNA
SRY	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
sex-determining	NN	O	O
gene	NN	O	O
SRY	NN	O	B-DNA
was	NN	O	O
screened	NN	O	O
for	NN	O	O
molecular	NN	O	O
alteration	NN	O	O
in	NN	O	O
an	NN	O	O
XY	NN	O	O
sex-reversed	NN	O	O
female	NN	O	O
by	NN	O	O
single-strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
technique	NN	O	O
.	NN	O	O

An	NN	O	O
A-to-G	NN	O	O
transition	NN	O	O
was	NN	O	O
detected	NN	O	O
which	NN	O	O
leads	NN	O	O
to	NN	O	O
an	NN	O	O
exchange	NN	O	O
of	NN	O	O
a	NN	O	O
tyrosine	NN	O	O
by	NN	O	O
a	NN	O	O
cysteine	NN	O	O
in	NN	O	O
the	NN	O	O
SRY	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
affected	NN	O	O
tyrosine	NN	O	O
residue	NN	O	O
located	NN	O	O
at	NN	O	O
the	NN	O	O
C	NN	O	B-protein
terminus	NN	O	I-protein
of	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
evolutionarily	NN	O	O
strongly	NN	O	O
conserved	NN	O	O
among	NN	O	O
the	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
containing	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
gel	NN	O	O
shift	NN	O	O
assay	NN	O	O
and	NN	O	O
peptide	NN	O	O
synthesis	NN	O	O
such	NN	O	O
a	NN	O	O
mutation	NN	O	O
is	NN	O	O
shown	NN	O	O
to	NN	O	O
abolish	NN	O	O
the	NN	O	O
SRY	NN	O	B-DNA
protein	NN	O	O
DNA	NN	O	O
binding	NN	O	O
ability	NN	O	O
.	NN	O	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
this	NN	O	O
particular	NN	O	O
amino	NN	O	O
acid	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
specificity	NN	O	O
is	NN	O	O
also	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
T-cell	NN	O	B-cell_type
activation	NN	O	O
by	NN	O	O
vanadate	NN	O	O
peroxide	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
phosphatases	NN	O	I-protein
.	NN	O	O

Rapid	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
key	NN	O	O
cellular	NN	O	O
proteins	NN	O	O
is	NN	O	O
a	NN	O	O
crucial	NN	O	O
event	NN	O	O
in	NN	O	O
the	NN	O	O
transduction	NN	O	O
of	NN	O	O
activation	NN	O	O
signals	NN	O	O
to	NN	O	O
T-lymphocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
regulatory	NN	O	O
role	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
phosphatases	NN	O	I-protein
(	NN	O	O
PTPases	NN	O	B-protein
)	NN	O	O
in	NN	O	O
this	NN	O	O
process	NN	O	O
was	NN	O	O
explored	NN	O	O
by	NN	O	O
studying	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
a	NN	O	O
powerful	NN	O	O
PTPase	NN	O	B-protein
inhibitor	NN	O	O
,	NN	O	O
vanadate	NN	O	O
peroxide	NN	O	O
(	NN	O	O
pervanadate	NN	O	O
)	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
cascade	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
human	NN	O	I-cell_line
leukaemic	NN	O	I-cell_line
T-cells	NN	O	I-cell_line
.	NN	O	O

Pervanadate	NN	O	O
induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
lck	NN	O	B-protein
and	NN	O	O
fyn	NN	O	B-protein
(	NN	O	O
4-	NN	O	O
and	NN	O	O
3-fold	NN	O	O
respectively	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
dramatic	NN	O	O
increase	NN	O	O
in	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
notably	NN	O	O
phospholipase	NN	O	B-protein
C	NN	O	I-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
.	NN	O	O

After	NN	O	O
this	NN	O	O
event	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
a	NN	O	O
rise	NN	O	O
in	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
concentration	NN	O	O
,	NN	O	O
corresponding	NN	O	O
to	NN	O	O
an	NN	O	O
influx	NN	O	O
.	NN	O	O

This	NN	O	O
effect	NN	O	O
required	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CD45	NN	O	B-protein
PTPase	NN	O	I-protein
and	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
CD45-deficient	NN	O	B-cell_line
variants	NN	O	I-cell_line
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
the	NN	O	O
CD45-negative	NN	O	B-cell_line
variant	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
pervanadate	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
was	NN	O	O
globally	NN	O	O
decreased	NN	O	O
and	NN	O	O
some	NN	O	O
phosphorylated	NN	O	O
substrates	NN	O	O
were	NN	O	O
specifically	NN	O	O
missing	NN	O	O
.	NN	O	O

Pervanadate	NN	O	O
also	NN	O	O
stimulated	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
accumulation	NN	O	O
of	NN	O	O
its	NN	O	O
mRNA	NN	O	B-RNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
several	NN	O	O
other	NN	O	O
hallmarks	NN	O	O
of	NN	O	O
T-lymphocyte	NN	O	B-cell_type
activation	NN	O	O
such	NN	O	O
as	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CD69	NN	O	O
antigen	NN	O	O
and	NN	O	O
the	NN	O	O
interleukin	NN	O	O
2	NN	O	O
receptor	NN	O	O
alpha-chain	NN	O	O
(	NN	O	O
CD25	NN	O	O
)	NN	O	O
.	NN	O	O

Pervanadate	NN	O	O
synergized	NN	O	O
with	NN	O	O
signals	NN	O	O
delivered	NN	O	O
by	NN	O	O
T-cell	NN	O	B-cell_type
antigen	NN	O	B-protein
receptor	NN	O	I-protein
engagement	NN	O	O
or	NN	O	O
by	NN	O	O
a	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
to	NN	O	O
induce	NN	O	O
interleukin	NN	O	O
2	NN	O	O
production	NN	O	O
.	NN	O	O

Pervanadate	NN	O	B-protein
activated	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

We	NN	O	O
thus	NN	O	O
conclude	NN	O	O
that	NN	O	O
PTPases	NN	O	B-protein
play	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
culminating	NN	O	O
in	NN	O	O
T-lymphocyte	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
appears	NN	O	O
sufficient	NN	O	O
per	NN	O	O
se	NN	O	O
to	NN	O	O
initiate	NN	O	O
a	NN	O	O
complete	NN	O	O
activation	NN	O	O
programme	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
via	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
requires	NN	O	O
a	NN	O	O
Raf	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
Ca2+	NN	O	O
influx	NN	O	O
.	NN	O	O

Functional	NN	O	O
synergy	NN	O	O
between	NN	O	O
Raf	NN	O	B-protein
and	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

Signals	NN	O	O
transduced	NN	O	O
via	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
activate	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
,	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
is	NN	O	O
critical	NN	O	O
to	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
many	NN	O	O
genes	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
differentiated	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
activator	NN	O	O
PMA	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
Ca2+	NN	O	O
ionophores	NN	O	O
mimics	NN	O	O
this	NN	O	O
process	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
two	NN	O	O
agents	NN	O	O
are	NN	O	O
often	NN	O	O
substituted	NN	O	O
for	NN	O	O
TCR	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
bypassing	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
identify	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
components	NN	O	O
involved	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
following	NN	O	O
TCR	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

TCR	NN	O	B-protein
signaling	NN	O	O
was	NN	O	O
triggered	NN	O	O
by	NN	O	O
treating	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
PHA	NN	O	B-protein
or	NN	O	O
anti-CD3	NN	O	B-protein
Abs	NN	O	I-protein
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
monitored	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
and/or	NN	O	O
by	NN	O	O
kappaB	NN	O	B-protein
-dependent	NN	O	O
reporter	NN	O	O
assays	NN	O	O
.	NN	O	O

Contrary	NN	O	O
to	NN	O	O
the	NN	O	O
idea	NN	O	O
that	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
high	NN	O	O
doses	NN	O	O
of	NN	O	O
staurosporine	NN	O	O
did	NN	O	O
not	NN	O	O
interfere	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
by	NN	O	O
PHA	NN	O	B-protein
,	NN	O	O
while	NN	O	O
the	NN	O	O
same	NN	O	O
dose	NN	O	O
of	NN	O	O
staurosporine	NN	O	O
completely	NN	O	O
blocked	NN	O	O
activation	NN	O	O
by	NN	O	O
PMA	NN	O	O
.	NN	O	O

PHA	NN	O	B-protein
-induced	NN	O	O
kappaB-dependent	NN	O	O
reporter	NN	O	O
activity	NN	O	O
was	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
effectively	NN	O	O
blocked	NN	O	O
by	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
form	NN	O	O
of	NN	O	O
Raf-1	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
for	NN	O	O
a	NN	O	O
Raf	NN	O	B-protein
kinase	NN	O	I-protein
.	NN	O	O

The	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
was	NN	O	O
also	NN	O	O
dependent	NN	O	O
on	NN	O	O
a	NN	O	O
Ca2+	NN	O	O
influx	NN	O	O
,	NN	O	O
because	NN	O	O
the	NN	O	O
Ca2+	NN	O	O
channel	NN	O	O
blocker	NN	O	O
,	NN	O	O
SK	NN	O	O
&	NN	O	O
F	NN	O	O
96365	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
other	NN	O	O
agents	NN	O	O
that	NN	O	O
prevented	NN	O	O
the	NN	O	O
Ca2+	NN	O	O
influx	NN	O	O
,	NN	O	O
inhibited	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
a	NN	O	O
constitutively	NN	O	O
active	NN	O	O
form	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
largely	NN	O	O
substituted	NN	O	O
for	NN	O	O
the	NN	O	O
Ca2+	NN	O	O
requirement	NN	O	O
and	NN	O	O
reversed	NN	O	O
the	NN	O	O
blockade	NN	O	O
by	NN	O	O
SK	NN	O	O
&	NN	O	O
F	NN	O	O
96365	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
these	NN	O	O
observations	NN	O	O
,	NN	O	O
coexpression	NN	O	O
of	NN	O	O
constitutively	NN	O	O
active	NN	O	O
forms	NN	O	O
of	NN	O	O
Raf-1	NN	O	B-protein
and	NN	O	O
calcineurin	NN	O	B-protein
synergistically	NN	O	O
induced	NN	O	O
kappaB	NN	O	B-protein
-dependent	NN	O	O
reporter	NN	O	O
activity	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
physiologically	NN	O	O
relevant	NN	O	O
functional	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
kinase	NN	O	B-protein
and	NN	O	O
the	NN	O	O
phosphatase	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Synergistic	NN	O	O
interactions	NN	O	O
between	NN	O	O
overlapping	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
serum	NN	O	B-protein
response	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
ELK-1	NN	O	B-protein
proteins	NN	O	I-protein
mediate	NN	O	O
both	NN	O	O
basal	NN	O	O
enhancement	NN	O	O
and	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
primate	NN	O	B-DNA
cytomegalovirus	NN	O	I-DNA
major	NN	O	I-DNA
immediate-early	NN	O	I-DNA
promoters	NN	O	I-DNA
in	NN	O	O
monocyte	NN	O	O
and	NN	O	O
T-lymphocyte	NN	O	B-cell_type
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Cytomegalovirus	NN	O	O
(	NN	O	O
CMV	NN	O	O
)	NN	O	O
infection	NN	O	O
is	NN	O	O
nonpermissive	NN	O	O
or	NN	O	O
persistent	NN	O	O
in	NN	O	O
many	NN	O	O
lymphoid	NN	O	B-cell_type
and	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
but	NN	O	O
can	NN	O	O
be	NN	O	O
activated	NN	O	O
in	NN	O	O
differentiated	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
elsewhere	NN	O	O
that	NN	O	O
both	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
immediate-early	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
MIE	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
infectious	NN	O	O
progeny	NN	O	O
virus	NN	O	O
expression	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
otherwise	NN	O	O
nonpermissive	NN	O	O
monocyte-like	NN	O	B-cell_line
U-937	NN	O	I-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
infected	NN	O	O
with	NN	O	O
either	NN	O	O
human	NN	O	O
CMV	NN	O	O
(	NN	O	O
HCMV	NN	O	O
)	NN	O	O
or	NN	O	O
simian	NN	O	O
CMV	NN	O	O
(	NN	O	O
SCMV	NN	O	O
)	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
the	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
.	NN	O	O

Two	NN	O	O
multicopy	NN	O	B-DNA
basal	NN	O	I-DNA
enhancer	NN	O	I-DNA
motifs	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
SCMV	NN	O	O
MIE	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
namely	NN	O	O
,	NN	O	O
11	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
16-bp	NN	O	O
cyclic	NN	O	B-DNA
AMP	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CRE	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
3	NN	O	O
copies	NN	O	O
of	NN	O	O
novel	NN	O	O
17-bp	NN	O	O
serum	NN	O	B-DNA
response	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
SRF	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
referred	NN	O	O
to	NN	O	O
as	NN	O	O
the	NN	O	O
SNE	NN	O	B-DNA
(	NN	O	O
SRF/NFkappaB-like	NN	O	B-DNA
element	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
four	NN	O	O
classical	NN	O	O
NFkappaB	NN	O	B-DNA
sites	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
HCMV	NN	O	O
version	NN	O	O
,	NN	O	O
contribute	NN	O	O
to	NN	O	O
TPA	NN	O	O
responsiveness	NN	O	O
in	NN	O	O
transient	NN	O	O
assays	NN	O	O
in	NN	O	O
monocyte	NN	O	B-cell_type
and	NN	O	I-cell_type
T-cell	NN	O	I-cell_type
types	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
SCMV	NN	O	B-DNA
SNE	NN	O	I-DNA
sites	NN	O	I-DNA
contain	NN	O	O
potential	NN	O	O
overlapping	NN	O	O
core	NN	O	O
recognition	NN	O	O
binding	NN	O	O
motifs	NN	O	O
for	NN	O	O
SRF	NN	O	B-protein
,	NN	O	O
Rel/NFkappaB	NN	O	B-protein
,	NN	O	O
ETS	NN	O	B-protein
,	NN	O	O
and	NN	O	O
YY1	NN	O	B-protein
class	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
but	NN	O	O
fail	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
either	NN	O	O
serum	NN	O	B-protein
or	NN	O	I-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
to	NN	O	O
evaluate	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
TPA	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
the	NN	O	O
SNE	NN	O	B-DNA
motifs	NN	O	I-DNA
and	NN	O	O
of	NN	O	O
a	NN	O	O
related	NN	O	O
16-bp	NN	O	O
SEE	NN	O	B-DNA
(	NN	O	O
SRF/ETS	NN	O	B-DNA
element	NN	O	I-DNA
)	NN	O	O
motif	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
HCMV	NN	O	B-DNA
and	NN	O	I-DNA
chimpanzee	NN	O	I-DNA
CMV	NN	O	I-DNA
MIE	NN	O	I-DNA
enhancers	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
functional	NN	O	O
responses	NN	O	O
and	NN	O	O
protein	NN	O	O
binding	NN	O	O
properties	NN	O	O
of	NN	O	O
multimerized	NN	O	B-DNA
wild-type	NN	O	I-DNA
and	NN	O	I-DNA
mutant	NN	O	I-DNA
elements	NN	O	I-DNA
added	NN	O	O
upstream	NN	O	O
to	NN	O	O
the	NN	O	O
SCMV	NN	O	B-DNA
MIE	NN	O	I-DNA
or	NN	O	O
simian	NN	O	B-DNA
virus	NN	O	I-DNA
40	NN	O	I-DNA
minimal	NN	O	I-DNA
promoter	NN	O	I-DNA
regions	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
U-937	NN	O	B-cell_line
,	NN	O	O
K-562	NN	O	B-cell_line
,	NN	O	O
HL-60	NN	O	B-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
Jurkat	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Unlike	NN	O	O
classical	NN	O	O
NFkappaB	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
neither	NN	O	O
the	NN	O	O
SNE	NN	O	B-DNA
nor	NN	O	O
the	NN	O	O
SEE	NN	O	B-DNA
motif	NN	O	I-DNA
responded	NN	O	O
to	NN	O	O
phosphatase	NN	O	O
inhibition	NN	O	O
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
TPA	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
both	NN	O	O
CMV	NN	O	B-DNA
elements	NN	O	I-DNA
proved	NN	O	O
to	NN	O	O
involve	NN	O	O
synergistic	NN	O	O
interactions	NN	O	O
between	NN	O	O
the	NN	O	O
core	NN	O	O
SRF	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
CCATATATGG	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
adjacent	NN	O	O
inverted	NN	O	O
ETS	NN	O	B-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
(	NN	O	O
TTCC	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
correlated	NN	O	O
directly	NN	O	O
with	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
bound	NN	O	O
tripartite	NN	O	B-protein
complex	NN	O	I-protein
containing	NN	O	O
both	NN	O	O
the	NN	O	O
cellular	NN	O	O
SRF	NN	O	B-protein
and	NN	O	O
ELK-1	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

This	NN	O	O
protein	NN	O	B-protein
complex	NN	O	I-protein
was	NN	O	O
more	NN	O	O
abundant	NN	O	O
in	NN	O	O
U-937	NN	O	O
,	NN	O	O
K-562	NN	O	O
,	NN	O	O
and	NN	O	O
HeLa	NN	O	O
cell	NN	O	O
extracts	NN	O	O
than	NN	O	O
in	NN	O	O
Raji	NN	O	B-cell_line
,	NN	O	O
HF	NN	O	B-cell_line
,	NN	O	O
BALB/c	NN	O	B-cell_line
3T3	NN	O	I-cell_line
,	NN	O	O
or	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
altered	NN	O	O
only	NN	O	O
twofold	NN	O	O
after	NN	O	O
TPA	NN	O	O
treatment	NN	O	O
.	NN	O	O

A	NN	O	O
40-fold	NN	O	O
stimulation	NN	O	O
of	NN	O	O
chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
activity	NN	O	O
mediated	NN	O	O
by	NN	O	O
four	NN	O	O
tandem	NN	O	B-DNA
repeats	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
SNE	NN	O	B-DNA
could	NN	O	O
be	NN	O	O
induced	NN	O	O
within	NN	O	O
2	NN	O	O
h	NN	O	O
(	NN	O	O
and	NN	O	O
up	NN	O	O
to	NN	O	O
250-fold	NN	O	O
within	NN	O	O
6	NN	O	O
h	NN	O	O
)	NN	O	O
after	NN	O	O
addition	NN	O	O
of	NN	O	O
TPA	NN	O	O
in	NN	O	O
DNA-transfected	NN	O	B-cell_line
U-937	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
stimulation	NN	O	O
appeared	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
true	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
event	NN	O	O
rather	NN	O	O
than	NN	O	O
a	NN	O	O
differentiation	NN	O	O
response	NN	O	O
.	NN	O	O

Slight	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
SRF	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
compared	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
classical	NN	O	O
c-Fos	NN	O	B-DNA
promoter	NN	O	I-DNA
serum	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
together	NN	O	O
with	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
spacing	NN	O	O
between	NN	O	O
the	NN	O	O
SRF	NN	O	B-DNA
and	NN	O	I-DNA
ETS	NN	O	I-DNA
motifs	NN	O	I-DNA
,	NN	O	O
appear	NN	O	O
to	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
inability	NN	O	O
of	NN	O	O
the	NN	O	O
SCMV	NN	O	B-DNA
SNEs	NN	O	I-DNA
to	NN	O	O
respond	NN	O	O
to	NN	O	O
serum	NN	O	O
induction	NN	O	O
.	NN	O	O

-DOCSTART-	O

T-cell	NN	O	B-cell_type
-directed	NN	O	O
TAL-1	NN	O	B-protein
expression	NN	O	O
induces	NN	O	O
T-cell	NN	O	O
malignancies	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
TAL-1	NN	O	B-DNA
gene	NN	O	I-DNA
specifies	NN	O	O
for	NN	O	O
a	NN	O	O
basic	NN	O	O
domain-helix-loop-helix	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
normal	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

In	NN	O	O
human	NN	O	O
pathology	NN	O	O
,	NN	O	O
the	NN	O	O
TAL-1	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
high	NN	O	O
percentage	NN	O	O
of	NN	O	O
T-cell	NN	O	B-cell_type
acute	NN	O	O
lymphoblastic	NN	O	O
leukemias	NN	O	O
in	NN	O	O
the	NN	O	O
pediatric	NN	O	O
age	NN	O	O
range	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
established	NN	O	O
whether	NN	O	O
the	NN	O	O
expression	NN	O	O
has	NN	O	O
a	NN	O	O
causal	NN	O	O
role	NN	O	O
in	NN	O	O
oncogenesis	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
phenotype	NN	O	O
of	NN	O	O
mouse	NN	O	B-cell_line
transgenic	NN	O	I-cell_line
lines	NN	O	I-cell_line
obtained	NN	O	O
by	NN	O	O
inducing	NN	O	O
tal-1	NN	O	B-protein
protein	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
using	NN	O	O
the	NN	O	O
LCK	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
survival	NN	O	O
rate	NN	O	O
of	NN	O	O
tal-1	NN	O	B-DNA
transgenic	NN	O	O
animals	NN	O	O
was	NN	O	O
much	NN	O	O
lower	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
control	NN	O	O
mice	NN	O	O
.	NN	O	O

Histopathological	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
lymphomas	NN	O	O
of	NN	O	O
T-cell	NN	O	O
type	NN	O	O
,	NN	O	O
often	NN	O	O
comprising	NN	O	O
a	NN	O	O
minor	NN	O	O
B-cell	NN	O	O
component	NN	O	O
.	NN	O	O

Some	NN	O	O
mice	NN	O	O
showed	NN	O	O
marked	NN	O	O
splenic	NN	O	O
lymphocyte	NN	O	O
depletion	NN	O	O
.	NN	O	O

Primary	NN	O	B-cell_line
lymphocyte	NN	O	I-cell_line
cultures	NN	O	I-cell_line
showed	NN	O	O
partial	NN	O	O
independence	NN	O	O
from	NN	O	O
exogenous	NN	O	O
growth	NN	O	O
stimuli	NN	O	O
and	NN	O	O
increased	NN	O	O
resistance	NN	O	O
to	NN	O	O
low-serum	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
unravel	NN	O	O
the	NN	O	O
tal-1	NN	O	B-DNA
oncogenic	NN	O	O
potential	NN	O	O
,	NN	O	O
a	NN	O	O
strain	NN	O	O
of	NN	O	O
tal-1	NN	O	B-DNA
transgenic	NN	O	O
mice	NN	O	O
was	NN	O	O
crossbred	NN	O	O
with	NN	O	O
p53-/-	NN	O	O
mice	NN	O	O
;	NN	O	O
the	NN	O	O
survival	NN	O	O
rate	NN	O	O
in	NN	O	O
these	NN	O	O
animals	NN	O	O
was	NN	O	O
reduced	NN	O	O
by	NN	O	O
more	NN	O	O
than	NN	O	O
one-half	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
tal-1	NN	O	B-DNA
mice	NN	O	O
,	NN	O	O
and	NN	O	O
histopathological	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
exclusively	NN	O	O
T-cell	NN	O	B-cell_type
lymphomas	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
TAL-1	NN	O	B-protein
,	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
per	NN	O	O
se	NN	O	O
a	NN	O	O
potent	NN	O	O
oncogene	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
exert	NN	O	O
a	NN	O	O
key	NN	O	O
leukemogenetic	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
T-cell	NN	O	B-cell_type
acute	NN	O	O
lymphoblastic	NN	O	O
leukemias	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tissue	NN	O	O
and	NN	O	O
cell-type	NN	O	O
specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
tuberous	NN	O	B-DNA
sclerosis	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
TSC2	NN	O	B-DNA
,	NN	O	O
in	NN	O	O
human	NN	O	O
tissues	NN	O	O
.	NN	O	O

TSC2	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
gene	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
16p13.3	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
the	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
neurocutaneous	NN	O	O
disorder	NN	O	O
,	NN	O	O
tuberous	NN	O	O
sclerosis	NN	O	O
complex	NN	O	O
(	NN	O	O
TSC	NN	O	O
)	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
a	NN	O	O
partial	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
from	NN	O	O
the	NN	O	O
cloned	NN	O	O
TSC2	NN	O	B-DNA
and	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
methodology	NN	O	O
,	NN	O	O
we	NN	O	O
constructed	NN	O	O
a	NN	O	O
digoxigenin-labeled	NN	O	B-DNA
complementary	NN	O	I-DNA
DNA	NN	O	I-DNA
probe	NN	O	I-DNA
to	NN	O	O
examine	NN	O	O
TSC2	NN	O	B-DNA
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
autopsy-	NN	O	O
or	NN	O	O
biopsy-derived	NN	O	O
human	NN	O	O
tissues	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

TSC2	NN	O	B-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
was	NN	O	O
widely	NN	O	O
expressed	NN	O	O
in	NN	O	O
various	NN	O	O
cell	NN	O	O
types	NN	O	O
throughout	NN	O	O
the	NN	O	O
body	NN	O	O
,	NN	O	O
including	NN	O	O
epithelia	NN	O	O
,	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
cells	NN	O	O
with	NN	O	O
endocrine	NN	O	O
functions	NN	O	O
,	NN	O	O
e.g.	NN	O	O
,	NN	O	O
adrenal	NN	O	O
cortex	NN	O	O
and	NN	O	O
anterior	NN	O	O
pituitary	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
prominently	NN	O	O
and	NN	O	O
selectively	NN	O	O
(	NN	O	O
within	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
)	NN	O	O
expressed	NN	O	O
in	NN	O	O
pyramidal	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
cerebral	NN	O	O
cortex	NN	O	O
and	NN	O	O
other	NN	O	O
motor	NN	O	B-cell_type
neurons	NN	O	I-cell_type
,	NN	O	O
e.g.	NN	O	O
,	NN	O	O
in	NN	O	O
spinal	NN	O	O
cord	NN	O	O
and	NN	O	O
brainstem	NN	O	O
nuclei	NN	O	O
.	NN	O	O

Visceral	NN	O	O
TSC2	NN	O	B-DNA
expression	NN	O	O
was	NN	O	O
comparable	NN	O	O
in	NN	O	O
autopsy	NN	O	O
tissues	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
and	NN	O	O
without	NN	O	O
TSC	NN	O	O
;	NN	O	O
TSC2	NN	O	B-DNA
messenger	NN	O	O
RNA	NN	O	O
expression	NN	O	O
was	NN	O	O
most	NN	O	O
prominent	NN	O	O
in	NN	O	O
cells	NN	O	O
with	NN	O	O
a	NN	O	O
rapid	NN	O	O
mitotic	NN	O	O
rate	NN	O	O
and	NN	O	O
turnover	NN	O	O
,	NN	O	O
e.g.	NN	O	O
,	NN	O	O
epithelia	NN	O	O
and	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
with	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
pyramidal	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
other	NN	O	O
neurons	NN	O	O
being	NN	O	O
an	NN	O	O
obvious	NN	O	O
exception	NN	O	O
,	NN	O	O
and/or	NN	O	O
in	NN	O	O
cells	NN	O	O
with	NN	O	O
important	NN	O	O
secretory/transport	NN	O	O
functions	NN	O	O
.	NN	O	O

This	NN	O	O
widespread	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
TSC2	NN	O	B-DNA
gene	NN	O	I-DNA
supports	NN	O	O
the	NN	O	O
view	NN	O	O
that	NN	O	O
it	NN	O	O
encodes	NN	O	O
a	NN	O	O
protein	NN	O	O
vital	NN	O	O
to	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
metabolism	NN	O	O
or	NN	O	O
one	NN	O	O
that	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
tumor/growth	NN	O	O
suppressor	NN	O	O
.	NN	O	O

-DOCSTART-	O

Apoptosis	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
differential	NN	O	O
activity	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
and	NN	O	O
IL-1beta-converting	NN	O	B-protein
enzyme	NN	O	I-protein
family	NN	O	I-protein
protease	NN	O	O
inhibitors	NN	O	O
on	NN	O	O
glucocorticoid-	NN	O	O
and	NN	O	O
Fas-mediated	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

Fas	NN	O	B-protein
-mediated	NN	O	O
apoptosis	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Restimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
blasts	NN	O	I-cell_type
up-regulates	NN	O	O
Fas	NN	O	B-protein
and	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	O
expression	NN	O	O
,	NN	O	O
with	NN	O	O
subsequent	NN	O	O
interaction	NN	O	O
leading	NN	O	O
to	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
in	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
blocks	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
many	NN	O	O
stimuli	NN	O	O
,	NN	O	O
but	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
-mediated	NN	O	O
killing	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
consistently	NN	O	O
observed	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
behavior	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
in	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
blasts	NN	O	I-cell_type
were	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
with	NN	O	O
Bcl-2	NN	O	B-protein
and	NN	O	O
related	NN	O	O
gene	NN	O	B-protein
products	NN	O	I-protein
to	NN	O	O
determine	NN	O	O
the	NN	O	O
effect	NN	O	O
on	NN	O	O
apoptotic	NN	O	O
signaling	NN	O	O
.	NN	O	O

Transient	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
in	NN	O	O
mouse	NN	O	O
and	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
blasts	NN	O	I-cell_type
did	NN	O	O
not	NN	O	O
block	NN	O	O
Fas	NN	O	B-protein
-mediated	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
whereas	NN	O	O
etoposide-	NN	O	O
and	NN	O	O
glucocorticoid-induced	NN	O	O
cytotoxicity	NN	O	O
was	NN	O	O
potently	NN	O	O
inhibited	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
Bcl-xL	NN	O	B-protein
and	NN	O	O
adenovirus	NN	O	B-protein
E1B	NN	O	I-protein
19K	NN	O	I-protein
did	NN	O	O
not	NN	O	O
interfere	NN	O	O
with	NN	O	O
anti-	NN	O	O
Fas	NN	O	B-protein
killing	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
interleukin-1beta-converting	NN	O	O
enzyme	NN	O	O
family	NN	O	O
protease	NN	O	O
inhibitors	NN	O	O
Ac-DEVD-CHO	NN	O	O
and	NN	O	O
CrmA	NN	O	O
blocked	NN	O	O
Fas	NN	O	B-protein
-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
use	NN	O	O
distinct	NN	O	O
apoptosis	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
with	NN	O	O
differential	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
Bcl-2	NN	O	B-protein
and	NN	O	O
interleukin-1beta-converting	NN	O	O
enzyme	NN	O	O
family	NN	O	O
protease	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

Since	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
normally	NN	O	O
express	NN	O	O
Bcl-2	NN	O	B-protein
and	NN	O	O
Bcl-xL	NN	O	B-protein
following	NN	O	O
activation	NN	O	O
,	NN	O	O
their	NN	O	O
inability	NN	O	O
to	NN	O	O
block	NN	O	O
Fas	NN	O	B-protein
-mediated	NN	O	O
apoptosis	NN	O	O
may	NN	O	O
allow	NN	O	O
for	NN	O	O
the	NN	O	O
elimination	NN	O	O
of	NN	O	O
self-reactive	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
appropriate	NN	O	O
regulation	NN	O	O
of	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Increased	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
transcription	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
ciprofloxacin	NN	O	O
.	NN	O	O

The	NN	O	O
fluoroquinolone	NN	O	O
antibiotic	NN	O	O
,	NN	O	O
ciprofloxacin	NN	O	O
(	NN	O	O
cipro	NN	O	O
)	NN	O	O
,	NN	O	O
induces	NN	O	O
hyperproduction	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
in	NN	O	O
stimulated	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
investigation	NN	O	O
an	NN	O	O
enhanced	NN	O	O
and	NN	O	O
prolonged	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
response	NN	O	O
was	NN	O	O
also	NN	O	O
detected	NN	O	O
in	NN	O	O
both	NN	O	O
stimulated	NN	O	O
(	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
mitogens	NN	O	I-protein
or	NN	O	O
alloantigens	NN	O	B-protein
)	NN	O	O
murine	NN	O	B-cell_type
splenocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
the	NN	O	O
stimulated	NN	O	O
murine	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
EL-4	NN	O	I-cell_line
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
ciprofloxacin	NN	O	O
(	NN	O	O
5-80	NN	O	O
micrograms/ml	NN	O	O
)	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
control	NN	O	O
cells	NN	O	O
without	NN	O	O
antibiotics	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
was	NN	O	O
inhibited	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
were	NN	O	O
unaffected	NN	O	O
at	NN	O	O
24	NN	O	O
h	NN	O	O
and	NN	O	O
only	NN	O	O
slightly	NN	O	O
upregulated	NN	O	O
at	NN	O	O
48	NN	O	O
and	NN	O	O
72	NN	O	O
h	NN	O	O
of	NN	O	O
culture	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
splenocytes	NN	O	I-cell_type
incubated	NN	O	O
with	NN	O	O
cipro	NN	O	O
(	NN	O	O
20	NN	O	O
micrograms/ml	NN	O	O
)	NN	O	O
.	NN	O	O

EL-4	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
plasmid	NN	O	O
containing	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
enhancer	NN	O	B-DNA
region	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
the	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
revealed	NN	O	O
that	NN	O	O
cipro	NN	O	O
enhanced	NN	O	O
IL-2	NN	O	O
gene	NN	O	O
induction	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
EL-4	NN	O	B-cell_type
cells	NN	O	I-cell_type
incubated	NN	O	O
with	NN	O	O
ciprofloxacin	NN	O	O
showed	NN	O	O
an	NN	O	O
early	NN	O	O
peak	NN	O	O
and	NN	O	O
more	NN	O	O
activated	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
NFAT-1	NN	O	B-protein
)	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
control	NN	O	O
cells	NN	O	O
without	NN	O	O
antibiotics	NN	O	O
.	NN	O	O

Cipro	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
AP-1	NN	O	B-protein
or	NN	O	O
NFIL-2A	NN	O	B-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
cipro	NN	O	O
inhibited	NN	O	O
IFN-gamma	NN	O	B-protein
synthesis	NN	O	O
,	NN	O	O
but	NN	O	O
enhanced	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
means	NN	O	O
of	NN	O	O
influencing	NN	O	O
NFAT-1	NN	O	B-protein
and	NN	O	O
causing	NN	O	O
an	NN	O	O
increased	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
Tax	NN	O	B-protein
associates	NN	O	O
with	NN	O	O
and	NN	O	O
is	NN	O	O
negatively	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B2	NN	O	I-protein
p100	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
:	NN	O	O
implications	NN	O	O
for	NN	O	O
viral	NN	O	O
latency	NN	O	O
.	NN	O	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
is	NN	O	O
the	NN	O	O
etiologic	NN	O	O
agent	NN	O	O
of	NN	O	O
the	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
,	NN	O	O
an	NN	O	O
aggressive	NN	O	O
and	NN	O	O
often	NN	O	O
fatal	NN	O	O
malignancy	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_line
human	NN	O	I-cell_line
CD4	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

HTLV-I	NN	O	O
encodes	NN	O	O
an	NN	O	O
essential	NN	O	O
40-kDa	NN	O	B-protein
protein	NN	O	I-protein
termed	NN	O	O
Tax	NN	O	B-protein
that	NN	O	O
not	NN	O	O
only	NN	O	O
transactivates	NN	O	O
the	NN	O	O
long	NN	O	O
terminal	NN	O	O
repeat	NN	O	O
of	NN	O	O
this	NN	O	O
retrovirus	NN	O	O
but	NN	O	O
also	NN	O	O
induces	NN	O	O
an	NN	O	O
array	NN	O	O
of	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Tax	NN	O	B-protein
-mediated	NN	O	O
transformation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
likely	NN	O	O
involves	NN	O	O
the	NN	O	O
deregulated	NN	O	O
expression	NN	O	O
of	NN	O	O
various	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
that	NN	O	O
normally	NN	O	O
regulate	NN	O	O
lymphocyte	NN	O	B-cell_type
growth	NN	O	O
produced	NN	O	O
by	NN	O	O
altered	NN	O	O
activity	NN	O	O
of	NN	O	O
various	NN	O	O
endogenous	NN	O	O
host	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
Tax	NN	O	B-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
modulating	NN	O	O
the	NN	O	O
expression	NN	O	O
or	NN	O	O
activity	NN	O	O
of	NN	O	O
various	NN	O	O
host	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
and	NN	O	O
CREB/ATF	NN	O	B-protein
families	NN	O	I-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
HEB-1	NN	O	B-protein
and	NN	O	O
p67SRF	NN	O	B-protein
.	NN	O	O

An	NN	O	O
additional	NN	O	O
distinguishing	NN	O	O
characteristic	NN	O	O
of	NN	O	O
HTLV-I	NN	O	O
infection	NN	O	O
is	NN	O	O
the	NN	O	O
profound	NN	O	O
state	NN	O	O
of	NN	O	O
viral	NN	O	O
latency	NN	O	O
that	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
circulating	NN	O	O
primary	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
HTLV-I	NN	O	O
Tax	NN	O	B-protein
can	NN	O	O
physically	NN	O	O
associate	NN	O	O
with	NN	O	O
p100	NN	O	B-protein
,	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
Rel-related	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B2	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
both	NN	O	O
in	NN	O	O
transfected	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
HTLV-I-infected	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
physical	NN	O	O
interaction	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
with	NN	O	O
p100	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HTLV-I	NN	O	O
and	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeats	NN	O	I-DNA
,	NN	O	O
reflecting	NN	O	O
p100	NN	O	B-protein
-mediated	NN	O	O
cytoplasmic	NN	O	O
sequestration	NN	O	O
of	NN	O	O
the	NN	O	O
normally	NN	O	O
nuclearly	NN	O	O
expressed	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
mutant	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
that	NN	O	O
selectively	NN	O	O
fails	NN	O	O
to	NN	O	O
activate	NN	O	O
nuclear	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
expression	NN	O	O
does	NN	O	O
not	NN	O	O
associate	NN	O	O
with	NN	O	O
p100	NN	O	B-protein
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Rhabdomyosarcomas	NN	O	O
do	NN	O	O
not	NN	O	O
contain	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
domains	NN	O	O
of	NN	O	O
myogenic	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Skeletal	NN	O	O
myogenesis	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
MyoD	NN	O	B-protein
,	NN	O	O
myogenin	NN	O	B-protein
,	NN	O	O
myf5	NN	O	B-protein
,	NN	O	O
and	NN	O	O
myf6	NN	O	B-protein
)	NN	O	O
that	NN	O	O
are	NN	O	O
``	NN	O	B-protein
basic	NN	O	I-protein
helix-loop-helix	NN	O	I-protein
''	NN	O	I-protein
proteins	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
muscle-specific	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
promote	NN	O	O
their	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
after	NN	O	O
a	NN	O	O
mutation	NN	O	O
of	NN	O	O
Leu122	NN	O	O
to	NN	O	O
Arg	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
basic	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
MyoD	NN	O	B-protein
confers	NN	O	O
c-myc	NN	O	B-DNA
-like	NN	O	O
functional	NN	O	O
characteristics	NN	O	O
to	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
used	NN	O	O
single-strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
such	NN	O	O
mutations	NN	O	O
occur	NN	O	O
naturally	NN	O	O
in	NN	O	O
rhabdomyosarcomas	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
all	NN	O	O
the	NN	O	O
myogenic	NN	O	B-protein
factors	NN	O	I-protein
remain	NN	O	O
unaltered	NN	O	O
in	NN	O	O
rhabdomyosarcomas	NN	O	O
.	NN	O	O

Selection	NN	O	O
against	NN	O	O
such	NN	O	O
mutations	NN	O	O
may	NN	O	O
be	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
functional	NN	O	O
redundancy	NN	O	O
of	NN	O	O
these	NN	O	O
myogenic	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Function	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
ubiquitous	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
its	NN	O	O
properties	NN	O	O
seem	NN	O	O
to	NN	O	O
be	NN	O	O
most	NN	O	O
extensively	NN	O	O
exploited	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Among	NN	O	O
these	NN	O	O
properties	NN	O	O
are	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
's	NN	O	O
rapid	NN	O	O
posttranslational	NN	O	O
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
many	NN	O	O
pathogenic	NN	O	O
signals	NN	O	O
,	NN	O	O
its	NN	O	O
direct	NN	O	O
participation	NN	O	O
in	NN	O	O
cytoplasmic/nuclear	NN	O	O
signaling	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
potency	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
great	NN	O	O
variety	NN	O	O
of	NN	O	O
genes	NN	O	O
encoding	NN	O	O
immunologically	NN	O	B-protein
relevant	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
vertebrates	NN	O	O
,	NN	O	O
five	NN	O	O
distinct	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
subunits	NN	O	I-protein
are	NN	O	O
currently	NN	O	O
known	NN	O	O
which	NN	O	O
might	NN	O	O
extensively	NN	O	O
heterodimerize	NN	O	O
,	NN	O	O
thereby	NN	O	O
forming	NN	O	O
complexes	NN	O	O
with	NN	O	O
distinct	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
,	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
specificity	NN	O	O
,	NN	O	O
and	NN	O	O
cell	NN	O	O
type-	NN	O	O
and	NN	O	O
cell	NN	O	O
stage-specific	NN	O	O
distribution	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
dimers	NN	O	I-protein
is	NN	O	O
tightly	NN	O	O
controlled	NN	O	O
by	NN	O	O
accessory	NN	O	B-protein
proteins	NN	O	I-protein
called	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
subunits	NN	O	I-protein
of	NN	O	O
which	NN	O	O
there	NN	O	O
are	NN	O	O
also	NN	O	O
five	NN	O	O
different	NN	O	O
species	NN	O	O
currently	NN	O	O
known	NN	O	O
in	NN	O	O
vertebrates	NN	O	O
.	NN	O	O

I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
inhibit	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
prevent	NN	O	O
nuclear	NN	O	O
uptake	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

An	NN	O	O
exception	NN	O	O
is	NN	O	O
the	NN	O	O
Bcl-3	NN	O	B-protein
protein	NN	O	I-protein
which	NN	O	O
in	NN	O	O
addition	NN	O	O
can	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
activating	NN	O	I-protein
subunit	NN	O	I-protein
in	NN	O	O
th	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Other	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
rather	NN	O	O
involved	NN	O	O
in	NN	O	O
terminating	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
's	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

The	NN	O	O
intracellular	NN	O	O
events	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
inactivation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
i.e.	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
are	NN	O	O
complex	NN	O	O
.	NN	O	O

They	NN	O	O
involve	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
proteolytic	NN	O	O
reactions	NN	O	O
and	NN	O	O
seem	NN	O	O
to	NN	O	O
be	NN	O	O
controlled	NN	O	O
by	NN	O	O
the	NN	O	O
cells	NN	O	O
'	NN	O	O
redox	NN	O	O
status	NN	O	O
.	NN	O	O

Interference	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
or	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
may	NN	O	O
be	NN	O	O
beneficial	NN	O	O
in	NN	O	O
suppressing	NN	O	O
toxic/septic	NN	O	O
shock	NN	O	O
,	NN	O	O
graft-vs-host	NN	O	O
reactions	NN	O	O
,	NN	O	O
acute	NN	O	O
inflammatory	NN	O	O
reactions	NN	O	O
,	NN	O	O
acute	NN	O	O
phase	NN	O	O
response	NN	O	O
,	NN	O	O
and	NN	O	O
radiation	NN	O	O
damage	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
antioxidants	NN	O	O
and	NN	O	O
specific	NN	O	O
protease	NN	O	O
inhibitors	NN	O	O
may	NN	O	O
provide	NN	O	O
a	NN	O	O
pharmacological	NN	O	O
basis	NN	O	O
for	NN	O	O
interfering	NN	O	O
with	NN	O	O
these	NN	O	O
acute	NN	O	O
processes	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lack	NN	O	O
of	NN	O	O
T-cell-mediated	NN	O	O
recognition	NN	O	O
of	NN	O	O
the	NN	O	O
fusion	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
pml/RAR-alpha	NN	O	B-protein
hybrid	NN	O	I-protein
protein	NN	O	I-protein
by	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
previous	NN	O	O
studies	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
fusion	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
pml/RAR-alpha	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
expressed	NN	O	O
by	NN	O	O
acute	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
(	NN	O	I-cell_line
APL	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
can	NN	O	O
be	NN	O	O
specifically	NN	O	O
recognized	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
donor	NN	O	O
(	NN	O	O
D.	NN	O	O
E.	NN	O	O
)	NN	O	O
CD4	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
a	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
DR11-restricted	NN	O	O
fashion	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
here	NN	O	O
the	NN	O	O
results	NN	O	O
on	NN	O	O
the	NN	O	O
recognition	NN	O	O
of	NN	O	O
several	NN	O	O
pml/RAR-alpha	NN	O	O
peptides	NN	O	O
by	NN	O	O
APL	NN	O	O
patients	NN	O	O
expressing	NN	O	O
HLA	NN	O	B-protein
DR11	NN	O	I-protein
.	NN	O	O

The	NN	O	O
in	NN	O	O
vitro	NN	O	O
immunization	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
four	NN	O	O
patients	NN	O	O
in	NN	O	O
remission	NN	O	O
(	NN	O	O
S.R.	NN	O	O
,	NN	O	O
F.R.	NN	O	O
,	NN	O	O
M.M.	NN	O	O
,	NN	O	O
P.	NN	O	O
G.	NN	O	O
)	NN	O	O
with	NN	O	O
BCR1/25	NN	O	O
,	NN	O	O
a	NN	O	O
25-mer	NN	O	O
pml/	NN	O	O
RAR-alpha	NN	O	B-protein
,	NN	O	O
did	NN	O	O
not	NN	O	O
elicit	NN	O	O
either	NN	O	O
a	NN	O	O
polyclonal	NN	O	O
or	NN	O	O
a	NN	O	O
clonal	NN	O	O
immune	NN	O	O
response	NN	O	O
specific	NN	O	O
to	NN	O	O
the	NN	O	O
peptide	NN	O	O
.	NN	O	O

We	NN	O	O
then	NN	O	O
generated	NN	O	O
new	NN	O	O
donor	NN	O	O
anti-pml/RAR-alpha	NN	O	B-cell_line
CD4	NN	O	I-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
clones	NN	O	O
were	NN	O	O
tested	NN	O	O
for	NN	O	O
their	NN	O	O
recognition	NN	O	O
of	NN	O	O
BCR1/25	NN	O	O
.	NN	O	O

One	NN	O	O
clone	NN	O	O
(	NN	O	O
C3/5	NN	O	B-cell_line
,	NN	O	I-cell_line
CD3	NN	O	I-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
,	NN	O	I-cell_line
CD4	NN	O	I-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
,	NN	O	I-cell_line
CD8	NN	O	I-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
)	NN	O	O
was	NN	O	O
selected	NN	O	O
for	NN	O	O
further	NN	O	O
analysis	NN	O	O
.	NN	O	O

Clone	NN	O	B-cell_line
C3/5	NN	O	I-cell_line
showed	NN	O	O
specific	NN	O	O
proliferation	NN	O	O
,	NN	O	O
cytotoxicity	NN	O	O
,	NN	O	O
and	NN	O	O
cytokine	NN	O	O
(	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
)	NN	O	O
production	NN	O	O
when	NN	O	O
challenged	NN	O	O
with	NN	O	O
autologous	NN	O	B-cell_line
lymphoblastic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
pulsed	NN	O	O
with	NN	O	O
peptide	NN	O	O
BCR1/25	NN	O	O
.	NN	O	O

C3/5	NN	O	B-cell_line
cells	NN	O	I-cell_line
developed	NN	O	O
specific	NN	O	O
proliferation	NN	O	O
and	NN	O	O
cytotoxicity	NN	O	O
when	NN	O	O
challenged	NN	O	O
with	NN	O	O
peptide-pulsed	NN	O	B-cell_line
lymphoblastic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
four	NN	O	O
DR11	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
APL	NN	O	O
patients	NN	O	O
.	NN	O	O

APL	NN	O	B-cell_type
blasts	NN	O	I-cell_type
,	NN	O	O
available	NN	O	O
only	NN	O	O
from	NN	O	O
patients	NN	O	O
F.R.	NN	O	O
and	NN	O	O
P.G.	NN	O	O
,	NN	O	O
were	NN	O	O
not	NN	O	O
lysed	NN	O	O
by	NN	O	O
C3/5	NN	O	B-cell_line
and	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
present	NN	O	O
peptide	NN	O	O
BCR1/25	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
APL	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
IFN-gamma	NN	O	B-protein
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
and	NN	O	O
recognition	NN	O	O
by	NN	O	O
the	NN	O	O
C3/5	NN	O	B-cell_line
clone	NN	O	I-cell_line
.	NN	O	O

Since	NN	O	O
APL	NN	O	B-cell_line
cells	NN	O	I-cell_line
do	NN	O	O
not	NN	O	O
express	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
we	NN	O	O
tested	NN	O	O
in	NN	O	O
two	NN	O	O
donors	NN	O	O
(	NN	O	O
D.E.	NN	O	O
and	NN	O	O
C.H.R.	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
patients	NN	O	O
S.R.and	NN	O	O
P.G.whether	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
9-mer	NN	O	O
peptides	NN	O	O
(	NN	O	O
BCR1/9	NN	O	O
)	NN	O	O
would	NN	O	O
generate	NN	O	O
a	NN	O	O
CD8	NN	O	B-protein
/HLA	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
-restricted	NN	O	O
response	NN	O	O
.	NN	O	O

No	NN	O	O
peptide-specific	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
or	NN	O	O
clone	NN	O	O
could	NN	O	O
be	NN	O	O
generated	NN	O	O
from	NN	O	O
both	NN	O	O
donors	NN	O	O
and	NN	O	O
patients	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
are	NN	O	O
discussed	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
possible	NN	O	O
therapeutic	NN	O	O
approaches	NN	O	O
to	NN	O	O
the	NN	O	O
immunotherapy	NN	O	O
of	NN	O	O
APL	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
proximal	NN	O	O
regulatory	NN	O	O
element	NN	O	O
of	NN	O	O
the	NN	O	O
interferon-gamma	NN	O	B-DNA
promoter	NN	O	I-DNA
mediates	NN	O	O
selective	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
is	NN	O	O
produced	NN	O	O
by	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
certain	NN	O	O
subsets	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
the	NN	O	O
basis	NN	O	O
for	NN	O	O
its	NN	O	O
selective	NN	O	O
expression	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

Within	NN	O	O
the	NN	O	O
region	NN	O	O
between	NN	O	O
-108	NN	O	O
and	NN	O	O
-40	NN	O	O
base	NN	O	O
pairs	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
promoter	NN	O	O
are	NN	O	O
two	NN	O	O
conserved	NN	O	O
and	NN	O	O
essential	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
confer	NN	O	O
activation-specific	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
report	NN	O	O
describes	NN	O	O
studies	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
most	NN	O	O
proximal	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
is	NN	O	O
an	NN	O	O
important	NN	O	O
determinant	NN	O	O
of	NN	O	O
its	NN	O	O
restricted	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
proximal	NN	O	O
element	NN	O	O
is	NN	O	O
a	NN	O	O
composite	NN	O	O
site	NN	O	O
that	NN	O	O
binds	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
CREB/ATF	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
octamer	NN	O	B-protein
families	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Jun	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
activation-induced	NN	O	O
transcription	NN	O	O
and	NN	O	O
binds	NN	O	O
preferably	NN	O	O
as	NN	O	O
a	NN	O	O
heterodimer	NN	O	O
with	NN	O	O
ATF-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
CREB	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
dampen	NN	O	O
transcription	NN	O	O
from	NN	O	O
this	NN	O	O
element	NN	O	O
.	NN	O	O

The	NN	O	O
CpG	NN	O	O
dinucleotide	NN	O	O
in	NN	O	O
this	NN	O	O
element	NN	O	B-DNA
is	NN	O	O
selectively	NN	O	O
methylated	NN	O	O
in	NN	O	O
Th2	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
other	NN	O	O
cells	NN	O	O
that	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
methylation	NN	O	O
markedly	NN	O	O
reduces	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
DNA	NN	O	O
methylation	NN	O	O
and	NN	O	O
for	NN	O	O
binding	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
mediate	NN	O	O
or	NN	O	O
impede	NN	O	O
transcription	NN	O	O
,	NN	O	O
this	NN	O	O
element	NN	O	O
appears	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
controlling	NN	O	O
IFN-gamma	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD40	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
lipopolysaccharide	NN	O	O
and	NN	O	O
anti-IgM	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
persistent	NN	O	O
nuclear	NN	O	O
accumulation	NN	O	O
of	NN	O	O
RelB	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
stimulation	NN	O	O
on	NN	O	O
the	NN	O	O
activity	NN	O	O
and	NN	O	O
nuclear	NN	O	O
appearance	NN	O	O
of	NN	O	O
Rel/nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	I-protein
NF-kappaB	NN	O	I-protein
)	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
primary	NN	O	B-cell_type
murine	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
triggering	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
ligands	NN	O	I-protein
expressed	NN	O	O
on	NN	O	O
L	NN	O	B-cell_type
cells	NN	O	I-cell_type
led	NN	O	O
to	NN	O	O
strong	NN	O	O
activation	NN	O	O
of	NN	O	O
an	NN	O	O
NF-kappaB-controlled	NN	O	B-DNA
beta-globin	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
primary	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Analyses	NN	O	O
of	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
individual	NN	O	O
members	NN	O	O
of	NN	O	O
Rel	NN	O	B-protein
proteins	NN	O	I-protein
after	NN	O	O
CD40	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
showed	NN	O	O
a	NN	O	O
strong	NN	O	O
and	NN	O	O
long-lasting	NN	O	O
accumulation	NN	O	O
of	NN	O	O
RelB	NN	O	B-protein
and	NN	O	O
,	NN	O	O
less	NN	O	O
pronounced	NN	O	O
,	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
.	NN	O	O

LPS	NN	O	O
stimulation	NN	O	O
did	NN	O	O
not	NN	O	O
give	NN	O	O
rise	NN	O	O
to	NN	O	O
a	NN	O	O
persistent	NN	O	O
nuclear	NN	O	O
accumulation	NN	O	O
of	NN	O	O
RelB	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
nuclear	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
RelB	NN	O	B-protein
,	NN	O	O
accumulated	NN	O	O
after	NN	O	O
B	NN	O	O
cell	NN	O	O
receptor	NN	O	O
stimulation	NN	O	O
.	NN	O	O

CD40	NN	O	B-protein
induced	NN	O	O
not	NN	O	O
only	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
but	NN	O	O
also	NN	O	O
de	NN	O	O
novo	NN	O	O
synthesis	NN	O	O
of	NN	O	O
RelB	NN	O	B-RNA
RNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
.	NN	O	O

S107	NN	O	B-cell_line
plasmacytoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
express	NN	O	O
CD40	NN	O	B-protein
but	NN	O	O
are	NN	O	O
defective	NN	O	O
for	NN	O	O
the	NN	O	O
nuclear	NN	O	O
appearance	NN	O	O
of	NN	O	O
p50/p65-NF-kappaB	NN	O	B-protein
,	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
RelB	NN	O	B-protein
after	NN	O	O
CD40	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
S107	NN	O	B-cell_line
cells	NN	O	I-cell_line
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
relB	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
stimulation	NN	O	O
of	NN	O	O
nuclear	NN	O	O
RelB	NN	O	B-protein
translocation	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
exerts	NN	O	O
a	NN	O	O
long-lasting	NN	O	O
stimulatory	NN	O	O
effect	NN	O	O
on	NN	O	O
both	NN	O	O
the	NN	O	O
transcription	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
RelB	NN	O	B-protein
.	NN	O	O

Since	NN	O	O
LPS	NN	O	O
and	NN	O	O
anti-IgM	NN	O	B-protein
were	NN	O	O
unable	NN	O	O
to	NN	O	O
activate	NN	O	O
RelB	NN	O	B-protein
,	NN	O	O
CD40	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
trigger	NN	O	O
a	NN	O	O
special	NN	O	O
program	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and/or	NN	O	O
differentiation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
a	NN	O	O
leukocyte-specific	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
body	NN	O	O
.	NN	O	O

The	NN	O	O
nuclear	NN	O	O
body	NN	O	O
(	NN	O	O
NB	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
cellular	NN	O	O
organelle	NN	O	O
that	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
and	NN	O	O
viral	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
NB	NN	O	O
is	NN	O	O
also	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
antibodies	NN	O	B-protein
in	NN	O	O
the	NN	O	O
serum	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
the	NN	O	O
autoimmune	NN	O	O
disease	NN	O	O
primary	NN	O	O
biliary	NN	O	O
cirrhosis	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
serum	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
primary	NN	O	O
biliary	NN	O	O
cirrhosis	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
a	NN	O	O
cDNA	NN	O	O
encoding	NN	O	O
a	NN	O	O
novel	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
NB	NN	O	O
,	NN	O	O
a	NN	O	O
140-kDa	NN	O	B-protein
protein	NN	O	I-protein
designated	NN	O	O
Sp140	NN	O	B-protein
.	NN	O	O

The	NN	O	O
predicted	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
amino-terminal	NN	O	B-protein
portion	NN	O	I-protein
of	NN	O	O
Sp140	NN	O	B-protein
was	NN	O	O
similar	NN	O	O
to	NN	O	O
Sp100	NN	O	B-protein
,	NN	O	O
a	NN	O	O
previously	NN	O	O
identified	NN	O	O
NB	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
carboxyl	NN	O	B-protein
portion	NN	O	I-protein
of	NN	O	O
Sp140	NN	O	B-protein
contained	NN	O	O
a	NN	O	O
zinc-finger	NN	O	B-protein
domain	NN	O	I-protein
and	NN	O	O
a	NN	O	O
bromodomain	NN	O	B-protein
,	NN	O	O
motifs	NN	O	O
that	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
proteins	NN	O	O
regulating	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

High	NN	O	O
levels	NN	O	O
of	NN	O	O
Sp140	NN	O	B-RNA
mRNA	NN	O	I-RNA
were	NN	O	O
detected	NN	O	O
in	NN	O	O
human	NN	O	O
spleen	NN	O	O
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
other	NN	O	O
human	NN	O	O
tissues	NN	O	O
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
SP140	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
myeloid	NN	O	B-cell_line
precursor	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
HL60	NN	O	B-cell_line
and	NN	O	O
NB4	NN	O	B-cell_line
markedly	NN	O	O
increased	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
chemically	NN	O	O
induced	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Immunohistochemical	NN	O	O
techniques	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
SP140	NN	O	B-protein
localized	NN	O	O
to	NN	O	O
the	NN	O	O
NB	NN	O	O
in	NN	O	O
differentiated	NN	O	O
HL60	NN	O	B-cell_line
and	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
location	NN	O	O
of	NN	O	O
Sp140	NN	O	B-protein
in	NN	O	O
the	NN	O	O
NB	NN	O	O
,	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
in	NN	O	O
cells	NN	O	O
involved	NN	O	O
in	NN	O	O
host	NN	O	O
defense	NN	O	O
,	NN	O	O
suggest	NN	O	O
that	NN	O	O
Sp140	NN	O	B-protein
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
and	NN	O	O
viral	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
and	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
(	NN	O	O
GCS	NN	O	O
)	NN	O	O
profoundly	NN	O	O
inhibit	NN	O	O
several	NN	O	O
aspects	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
immunity	NN	O	O
largely	NN	O	O
through	NN	O	O
inhibition	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
posttranscriptional	NN	O	O
levels	NN	O	O
.	NN	O	O

GCS	NN	O	O
were	NN	O	O
also	NN	O	O
reported	NN	O	O
to	NN	O	O
act	NN	O	O
indirectly	NN	O	O
by	NN	O	O
inducing	NN	O	O
transforming	NN	O	O
growth	NN	O	B-protein
factor-beta	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
blocks	NN	O	O
T	NN	O	O
cell	NN	O	O
immunity	NN	O	O
.	NN	O	O

In	NN	O	O
exerting	NN	O	O
their	NN	O	O
antiproliferative	NN	O	O
effects	NN	O	O
,	NN	O	O
GCS	NN	O	O
diffuse	NN	O	O
into	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
where	NN	O	O
they	NN	O	O
bind	NN	O	O
their	NN	O	O
cytoplasmic	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
translocates	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
where	NN	O	O
it	NN	O	O
inhibits	NN	O	O
transcription	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
through	NN	O	O
direct	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
(	NN	O	O
GRE	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	O
region	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
or	NN	O	O
,	NN	O	O
alternatively	NN	O	O
,	NN	O	O
through	NN	O	O
antagonism	NN	O	O
of	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
their	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
on	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
GCS	NN	O	O
up-regulate	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
that	NN	O	O
correlates	NN	O	O
with	NN	O	O
enhanced	NN	O	O
cytokine	NN	O	B-protein
effects	NN	O	O
on	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
,	NN	O	O
we	NN	O	O
summarize	NN	O	O
the	NN	O	O
current	NN	O	O
state	NN	O	O
of	NN	O	O
knowledge	NN	O	O
of	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
GCS	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
phenomenon	NN	O	O
of	NN	O	O
steroid-induced	NN	O	O
rebound	NN	O	O
,	NN	O	O
which	NN	O	O
ensues	NN	O	O
upon	NN	O	O
GCS	NN	O	O
withdrawal	NN	O	O
.	NN	O	O

-DOCSTART-	O

Isolation	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
murine	NN	O	B-DNA
fra-1	NN	O	I-DNA
:	NN	O	O
induction	NN	O	O
mediated	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
and	NN	O	O
surface	NN	O	B-protein
Ig	NN	O	I-protein
is	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
dependent	NN	O	O
.	NN	O	O

The	NN	O	O
murine	NN	O	B-DNA
fra-1	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
encoding	NN	O	O
Fos-related	NN	O	B-protein
Ag	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
was	NN	O	O
isolated	NN	O	O
from	NN	O	O
a	NN	O	O
splenic	NN	O	B-DNA
cDNA	NN	O	I-DNA
library	NN	O	I-DNA
and	NN	O	O
sequenced	NN	O	O
.	NN	O	O

Murine	NN	O	B-DNA
fra-1	NN	O	I-DNA
was	NN	O	O
highly	NN	O	O
homologous	NN	O	O
to	NN	O	O
rat	NN	O	B-DNA
and	NN	O	I-DNA
human	NN	O	I-DNA
fra-1	NN	O	I-DNA
.	NN	O	O

Oligonucleotide	NN	O	O
primers	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
sequence	NN	O	I-DNA
were	NN	O	O
used	NN	O	O
to	NN	O	O
construct	NN	O	O
a	NN	O	O
quantitative	NN	O	O
reverse	NN	O	O
transcription-PCR	NN	O	O
assay	NN	O	O
for	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

B	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
stimulation	NN	O	O
via	NN	O	O
both	NN	O	O
CD40	NN	O	B-protein
and	NN	O	O
surface	NN	O	B-protein
Ig	NN	O	I-protein
(	NN	O	I-protein
sIg	NN	O	I-protein
)	NN	O	I-protein
receptors	NN	O	I-protein
substantially	NN	O	O
induced	NN	O	O
fra-1	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
and	NN	O	O
for	NN	O	O
both	NN	O	O
receptors	NN	O	O
,	NN	O	O
induction	NN	O	O
was	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
dependent	NN	O	O
.	NN	O	O

This	NN	O	O
contrasts	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
by	NN	O	O
both	NN	O	O
CD40	NN	O	B-protein
and	NN	O	O
sIg	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
PKC	NN	O	B-protein
independent	NN	O	O
and	NN	O	O
indicates	NN	O	O
that	NN	O	O
CD40	NN	O	B-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
signaling	NN	O	O
through	NN	O	O
PKC	NN	O	B-protein
or	NN	O	O
a	NN	O	O
closely	NN	O	O
related	NN	O	O
kinase	NN	O	B-protein
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
fra-1	NN	O	B-DNA
following	NN	O	O
engagement	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
did	NN	O	O
not	NN	O	O
require	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
PKC	NN	O	B-protein
-dependent	NN	O	O
linkage	NN	O	O
between	NN	O	O
CD40	NN	O	B-protein
and	NN	O	O
fra-1	NN	O	B-DNA
is	NN	O	O
direct	NN	O	O
.	NN	O	O

CD40	NN	O	B-protein
-mediated	NN	O	O
fra-1	NN	O	B-DNA
induction	NN	O	O
did	NN	O	O
require	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
CD40	NN	O	B-protein
,	NN	O	O
like	NN	O	O
sIg	NN	O	B-protein
,	NN	O	O
may	NN	O	O
employ	NN	O	O
PKC	NN	O	B-protein
in	NN	O	O
producing	NN	O	O
select	NN	O	O
outcomes	NN	O	O
,	NN	O	O
that	NN	O	O
individual	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
receptors	NN	O	I-protein
may	NN	O	O
signal	NN	O	O
downstream	NN	O	O
events	NN	O	O
via	NN	O	O
both	NN	O	O
PKC-dependent	NN	O	O
and	NN	O	O
PKC-independent	NN	O	O
pathways	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
multiple	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
may	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
closely	NN	O	O
related	NN	O	O
genes	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Pentoxifylline	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
infection	NN	O	O
with	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
.	NN	O	O

Cytokine	NN	O	O
dysregulation	NN	O	O
in	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
infection	NN	O	O
has	NN	O	O
been	NN	O	O
documented	NN	O	O
in	NN	O	O
numerous	NN	O	O
studies	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
cited	NN	O	O
as	NN	O	O
an	NN	O	O
important	NN	O	O
component	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
this	NN	O	O
retroviral	NN	O	O
infection	NN	O	O
.	NN	O	O

Pharmacological	NN	O	O
modification	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
dysregulation	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
as	NN	O	O
a	NN	O	O
therapeutic	NN	O	O
modality	NN	O	O
for	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

Dr.	NN	O	O
Dezube	NN	O	O
of	NN	O	O
Beth	NN	O	O
Israel	NN	O	O
Hospital	NN	O	O
(	NN	O	O
Boston	NN	O	O
)	NN	O	O
concisely	NN	O	O
reviews	NN	O	O
the	NN	O	O
state	NN	O	O
of	NN	O	O
our	NN	O	O
knowledge	NN	O	O
regarding	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
pentoxifylline	NN	O	O
on	NN	O	O
expression	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
,	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
known	NN	O	O
to	NN	O	O
influence	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
and	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
possible	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
of	NN	O	O
advanced	NN	O	O
infection	NN	O	O
with	NN	O	O
this	NN	O	O
virus	NN	O	O
.	NN	O	O

Pentoxifylline	NN	O	O
,	NN	O	O
a	NN	O	O
trisubstituted	NN	O	O
xanthine	NN	O	O
derivative	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
decrease	NN	O	O
blood	NN	O	O
viscosity	NN	O	O
and	NN	O	O
is	NN	O	O
reasonably	NN	O	O
well	NN	O	O
tolerated	NN	O	O
by	NN	O	O
most	NN	O	O
recipients	NN	O	O
of	NN	O	O
the	NN	O	O
drug	NN	O	O
.	NN	O	O

Results	NN	O	O
of	NN	O	O
preliminary	NN	O	O
studies	NN	O	O
,	NN	O	O
many	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
conducted	NN	O	O
by	NN	O	O
Dr.	NN	O	O
Dezube	NN	O	O
,	NN	O	O
suggest	NN	O	O
that	NN	O	O
use	NN	O	O
of	NN	O	O
this	NN	O	O
agent	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
antiretroviral	NN	O	O
compounds	NN	O	O
may	NN	O	O
prove	NN	O	O
useful	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
granulocyte-macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
cooperative	NN	O	O
binding	NN	O	O
of	NN	O	O
Elf-1	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
granulocyte-macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
GM-CSF	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
studied	NN	O	O
extensively	NN	O	O
as	NN	O	O
a	NN	O	O
model	NN	O	O
system	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
during	NN	O	O
T-lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
expressed	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
is	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
following	NN	O	O
activation	NN	O	O
through	NN	O	O
the	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
T-cell	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

A	NN	O	O
highly	NN	O	O
conserved	NN	O	O
19-bp	NN	O	O
element	NN	O	O
located	NN	O	O
immediately	NN	O	O
5	NN	O	O
'	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
TATA	NN	O	I-DNA
box	NN	O	I-DNA
(	NN	O	O
bp	NN	O	B-DNA
-34	NN	O	I-DNA
to	NN	O	I-DNA
-52	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
herein	NN	O	O
called	NN	O	O
purine	NN	O	B-DNA
box	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	O
PB1	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
a	NN	O	O
T-cell	NN	O	B-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
complex	NN	O	I-protein
and	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
following	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
PB1	NN	O	B-DNA
sequence	NN	O	O
motif	NN	O	O
is	NN	O	O
highly	NN	O	O
conserved	NN	O	O
in	NN	O	O
both	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
murine	NN	O	I-DNA
GM-CSF	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
PB1	NN	O	B-DNA
element	NN	O	I-DNA
alone	NN	O	O
confers	NN	O	O
inducibility	NN	O	O
on	NN	O	O
a	NN	O	O
heterologous	NN	O	O
promoter	NN	O	O
following	NN	O	O
transfection	NN	O	O
into	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
identify	NN	O	O
a	NN	O	O
major	NN	O	O
PB1	NN	O	B-protein
nuclear	NN	O	I-protein
protein-binding	NN	O	I-protein
complex	NN	O	I-protein
that	NN	O	O
is	NN	O	O
not	NN	O	O
present	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
is	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
following	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
PB1	NN	O	B-DNA
is	NN	O	O
composed	NN	O	O
of	NN	O	O
adjacent	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
Ets	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
mutagenesis	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
both	NN	O	O
the	NN	O	O
Ets	NN	O	B-DNA
and	NN	O	I-DNA
AP-1	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	O
PB1	NN	O	B-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
complex	NN	O	I-protein
and	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
element	NN	O	O
and	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Steroid-resistant	NN	O	O
asthma	NN	O	O
.	NN	O	O

Cellular	NN	O	O
mechanisms	NN	O	O
contributing	NN	O	O
to	NN	O	O
inadequate	NN	O	O
response	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
.	NN	O	O

The	NN	O	O
current	NN	O	O
study	NN	O	O
examined	NN	O	O
whether	NN	O	O
alterations	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
binding	NN	O	O
contribute	NN	O	O
to	NN	O	O
poor	NN	O	O
response	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
in	NN	O	O
asthma	NN	O	O
.	NN	O	O

29	NN	O	O
asthma	NN	O	O
patients	NN	O	O
with	NN	O	O
forced	NN	O	O
expiratory	NN	O	O
volume	NN	O	O
in	NN	O	O
1	NN	O	O
s	NN	O	O
(	NN	O	O
FEV1	NN	O	O
)	NN	O	O
<	NN	O	O
70	NN	O	O
%	NN	O	O
predicted	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

Patients	NN	O	O
were	NN	O	O
classified	NN	O	O
as	NN	O	O
steroid	NN	O	O
sensitive	NN	O	O
(	NN	O	O
SS	NN	O	O
)	NN	O	O
if	NN	O	O
their	NN	O	O
morning	NN	O	O
FEV1	NN	O	O
increased	NN	O	O
>	NN	O	O
30	NN	O	O
%	NN	O	O
after	NN	O	O
a	NN	O	O
1-wk	NN	O	O
course	NN	O	O
of	NN	O	O
oral	NN	O	O
prednisone	NN	O	O
20	NN	O	O
mg	NN	O	O
twice	NN	O	O
daily	NN	O	O
and	NN	O	O
steroid	NN	O	O
resistant	NN	O	O
(	NN	O	O
SR	NN	O	O
)	NN	O	O
if	NN	O	O
they	NN	O	O
failed	NN	O	O
to	NN	O	O
increase	NN	O	O
>	NN	O	O
15	NN	O	O
%	NN	O	O
.	NN	O	O

PBMC	NN	O	O
obtained	NN	O	O
from	NN	O	O
these	NN	O	O
two	NN	O	O
groups	NN	O	O
,	NN	O	O
17	NN	O	O
SR	NN	O	O
and	NN	O	O
12	NN	O	O
SS	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
12	NN	O	O
normal	NN	O	O
controls	NN	O	O
were	NN	O	O
analyzed.	NN	O	O
SR	NN	O	O
patients	NN	O	O
had	NN	O	O
two	NN	O	O
distinguishable	NN	O	O
GR	NN	O	B-protein
binding	NN	O	O
abnormalities	NN	O	O
:	NN	O	O
15	NN	O	O
of	NN	O	O
the	NN	O	O
17	NN	O	O
SR	NN	O	O
patients	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
GR	NN	O	B-protein
binding	NN	O	O
affinity	NN	O	O
,	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
SS	NN	O	O
patients	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.0001	NN	O	O
)	NN	O	O
and	NN	O	O
normal	NN	O	O
controls	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.0001	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
defect	NN	O	O
was	NN	O	O
localized	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
reverted	NN	O	O
to	NN	O	O
normal	NN	O	O
after	NN	O	O
48	NN	O	O
h	NN	O	O
in	NN	O	O
culture	NN	O	O
media	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
incubation	NN	O	O
with	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
sustained	NN	O	O
this	NN	O	O
abnormality	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
two	NN	O	O
SR	NN	O	O
patients	NN	O	O
had	NN	O	O
an	NN	O	O
abnormally	NN	O	O
low	NN	O	O
GR	NN	O	B-protein
number	NN	O	O
with	NN	O	O
normal	NN	O	O
binding	NN	O	O
affinity	NN	O	O
that	NN	O	O
was	NN	O	O
not	NN	O	O
limited	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
GR	NN	O	B-protein
number	NN	O	O
failed	NN	O	O
to	NN	O	O
normalize	NN	O	O
after	NN	O	O
incubation	NN	O	O
in	NN	O	O
media	NN	O	O
alone	NN	O	O
or	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
SR	NN	O	O
asthma	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
more	NN	O	O
than	NN	O	O
one	NN	O	O
abnormality	NN	O	O
,	NN	O	O
the	NN	O	O
majority	NN	O	O
related	NN	O	O
to	NN	O	O
a	NN	O	O
reversible	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
reduction	NN	O	O
in	NN	O	O
GR	NN	O	B-protein
binding	NN	O	O
affinity	NN	O	O
and	NN	O	O
the	NN	O	O
second	NN	O	O
related	NN	O	O
to	NN	O	O
an	NN	O	O
irreversible	NN	O	O
reduction	NN	O	O
in	NN	O	O
GR	NN	O	B-protein
number	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
may	NN	O	O
have	NN	O	O
important	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
design	NN	O	O
of	NN	O	O
alternative	NN	O	O
treatment	NN	O	O
approaches	NN	O	O
for	NN	O	O
recalcitrant	NN	O	O
asthma	NN	O	O
.	NN	O	O

-DOCSTART-	O

Prevalence	NN	O	O
of	NN	O	O
aneuploidy	NN	O	O
,	NN	O	O
overexpressed	NN	O	O
ER	NN	O	B-protein
,	NN	O	O
and	NN	O	O
overexpressed	NN	O	O
EGFR	NN	O	B-protein
in	NN	O	O
random	NN	O	O
breast	NN	O	O
aspirates	NN	O	O
of	NN	O	O
women	NN	O	O
at	NN	O	O
high	NN	O	O
and	NN	O	O
low	NN	O	O
risk	NN	O	O
for	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

Breast	NN	O	O
tissue	NN	O	O
biomarkers	NN	O	O
which	NN	O	O
accurately	NN	O	O
predict	NN	O	O
breast	NN	O	O
cancer	NN	O	O
development	NN	O	O
within	NN	O	O
a	NN	O	O
10	NN	O	O
year	NN	O	O
period	NN	O	O
in	NN	O	O
high	NN	O	O
risk	NN	O	O
women	NN	O	O
are	NN	O	O
needed	NN	O	O
but	NN	O	O
currently	NN	O	O
not	NN	O	O
available	NN	O	O
.	NN	O	O

We	NN	O	O
initiated	NN	O	O
this	NN	O	O
study	NN	O	O
to	NN	O	O
determine	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
breast	NN	O	O
tissue	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
women	NN	O	O
at	NN	O	O
high	NN	O	O
risk	NN	O	O
for	NN	O	O
breast	NN	O	O
cancer	NN	O	O
,	NN	O	O
and	NN	O	O
2	NN	O	O
)	NN	O	O
if	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
biomarker	NN	O	O
abnormalities	NN	O	O
is	NN	O	O
greater	NN	O	O
in	NN	O	O
high	NN	O	O
risk	NN	O	O
than	NN	O	O
in	NN	O	O
low	NN	O	O
risk	NN	O	O
women	NN	O	O
.	NN	O	O

Eligible	NN	O	O
high	NN	O	O
risk	NN	O	O
women	NN	O	O
were	NN	O	O
those	NN	O	O
with	NN	O	O
a	NN	O	O
first	NN	O	O
degree	NN	O	O
relative	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
,	NN	O	O
prior	NN	O	O
breast	NN	O	O
cancer	NN	O	O
,	NN	O	O
or	NN	O	O
precancerous	NN	O	O
mastopathy	NN	O	O
.	NN	O	O

Low	NN	O	O
risk	NN	O	O
women	NN	O	O
were	NN	O	O
those	NN	O	O
without	NN	O	O
these	NN	O	O
or	NN	O	O
other	NN	O	O
major	NN	O	O
identifiable	NN	O	O
risk	NN	O	O
factors	NN	O	O
.	NN	O	O

Ductal	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
obtained	NN	O	O
via	NN	O	O
random	NN	O	O
fine	NN	O	O
needle	NN	O	O
aspirations	NN	O	O
and	NN	O	O
cytologically	NN	O	O
classified	NN	O	O
.	NN	O	O

Biomarkers	NN	O	O
included	NN	O	O
DNA	NN	O	O
ploidy	NN	O	O
,	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
epidermal	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
EGFR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
prevalence	NN	O	O
of	NN	O	O
DNA	NN	O	O
aneuploidy	NN	O	O
was	NN	O	O
30	NN	O	O
%	NN	O	O
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
10	NN	O	O
%	NN	O	O
,	NN	O	O
and	NN	O	O
overexpression	NN	O	O
of	NN	O	O
EGFR	NN	O	B-protein
35	NN	O	O
%	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
206	NN	O	O
high	NN	O	O
risk	NN	O	O
women	NN	O	O
whose	NN	O	O
median	NN	O	O
10	NN	O	O
year	NN	O	O
Gail	NN	O	O
risk	NN	O	O
(	NN	O	O
projected	NN	O	O
probability	NN	O	O
)	NN	O	O
of	NN	O	O
developing	NN	O	O
breast	NN	O	O
cancer	NN	O	O
was	NN	O	O
4.5	NN	O	O
%	NN	O	O
.	NN	O	O

The	NN	O	O
prevalence	NN	O	O
of	NN	O	O
aneuploidy	NN	O	O
and	NN	O	O
overexpressed	NN	O	O
EGFR	NN	O	B-protein
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
in	NN	O	O
the	NN	O	O
high	NN	O	O
risk	NN	O	O
women	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
25	NN	O	O
low	NN	O	O
risk	NN	O	O
controls	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.002	NN	O	O
)	NN	O	O
,	NN	O	O
whose	NN	O	O
median	NN	O	O
10	NN	O	O
year	NN	O	O
Gail	NN	O	O
risk	NN	O	O
was	NN	O	O
0.7	NN	O	O
%	NN	O	O
.	NN	O	O

The	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
overexpression	NN	O	O
between	NN	O	O
high	NN	O	O
and	NN	O	O
low	NN	O	O
risk	NN	O	O
groups	NN	O	O
was	NN	O	O
not	NN	O	O
statistically	NN	O	O
significant	NN	O	O
(	NN	O	O
p	NN	O	O
=	NN	O	O
0.095	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
low	NN	O	O
prevalence	NN	O	O
of	NN	O	O
overexpressed	NN	O	O
ER	NN	O	B-protein
and	NN	O	O
the	NN	O	O
small	NN	O	O
number	NN	O	O
of	NN	O	O
controls	NN	O	O
.	NN	O	O

A	NN	O	O
significant	NN	O	O
difference	NN	O	O
was	NN	O	O
noted	NN	O	O
in	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
abnormal	NN	O	O
biomarkers	NN	O	O
between	NN	O	O
the	NN	O	O
high	NN	O	O
risk	NN	O	O
and	NN	O	O
low	NN	O	O
risk	NN	O	O
women	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
large	NN	O	O
prospective	NN	O	O
trial	NN	O	O
is	NN	O	O
needed	NN	O	O
to	NN	O	O
determine	NN	O	O
if	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
of	NN	O	O
these	NN	O	O
biomarkers	NN	O	O
,	NN	O	O
is	NN	O	O
predictive	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
nuclear	NN	O	B-RNA
factor	NN	O	I-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
subunit	NN	O	I-RNA
p65	NN	O	I-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
in	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-cell_line
human	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
treated	NN	O	O
with	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
.	NN	O	O

Lipopolysaccharide	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
potent	NN	O	O
trigger	NN	O	O
substances	NN	O	O
for	NN	O	O
monocytes	NN	O	O
and	NN	O	O
macrophages	NN	O	O
causing	NN	O	O
secretion	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
such	NN	O	O
as	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
interleukin-1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
factors	NN	O	O
involved	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
these	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
still	NN	O	O
unknown	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
;	NN	O	I-protein
heterodimer	NN	O	I-protein
of	NN	O	I-protein
p50	NN	O	I-protein
and	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
have	NN	O	O
been	NN	O	O
suggested	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
gene	NN	O	O
transcription	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
when	NN	O	O
monocytes	NN	O	B-cell_type
are	NN	O	O
stimulated	NN	O	O
with	NN	O	O
lipopolysaccharide	NN	O	O
.	NN	O	O

Nonsteroidal	NN	O	O
anti-inflammatory	NN	O	O
drugs	NN	O	O
such	NN	O	O
as	NN	O	O
salicylates	NN	O	O
have	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
treat	NN	O	O
symptoms	NN	O	O
of	NN	O	O
inflammation	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
new	NN	O	O
mechanism	NN	O	O
of	NN	O	O
drug	NN	O	O
action	NN	O	O
was	NN	O	O
suggested	NN	O	O
recently	NN	O	O
.	NN	O	O

Salicylates	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
inhibit	NN	O	O
lipopolysaccharide-induced	NN	O	O
gene	NN	O	O
transcription	NN	O	O
via	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
preventing	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibitor	NN	O	I-protein
``	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
``	NN	O	O
,	NN	O	O
blocking	NN	O	O
the	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
into	NN	O	O
the	NN	O	O
nuclear	NN	O	O
compartment	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
subunit	NN	O	O
involved	NN	O	O
in	NN	O	O
this	NN	O	O
mechanism	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
defined	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
salicylates	NN	O	O
affect	NN	O	O
cytokine	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
active	NN	O	O
and	NN	O	O
inactive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-RNA
B	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
in	NN	O	O
Porphyromonas	NN	O	O
gingivalis	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-cell_line
human	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
assessed	NN	O	O
.	NN	O	O

High	NN	O	O
doses	NN	O	O
of	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
suppressed	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	B-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
suppression	NN	O	O
of	NN	O	O
total	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
p50	NN	O	B-protein
on	NN	O	O
tissue	NN	O	O
oligonucleotide	NN	O	O
had	NN	O	O
no	NN	O	O
effects	NN	O	O
on	NN	O	O
lipopolysaccharide-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
subunit	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
salicylate	NN	O	O
treatment	NN	O	O
and	NN	O	O
highlight	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
salicylate	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Glucocorticoid-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
RANTES	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
chemokine	NN	O	B-protein
RANTES	NN	O	B-protein
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
allergic	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
including	NN	O	O
asthma	NN	O	O
and	NN	O	O
rhinitis	NN	O	O
which	NN	O	O
are	NN	O	O
frequently	NN	O	O
treated	NN	O	O
with	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
dexamethasone	NN	O	O
dramatically	NN	O	O
inhibited	NN	O	O
RANTES	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
dose	NN	O	O
dependently	NN	O	O
in	NN	O	O
anti-CD3	NN	O	B-cell_line
activated	NN	O	I-cell_line
Hut-78	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
human	NN	O	B-cell_type
PBMCs	NN	O	I-cell_type
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
RANTES	NN	O	B-protein
expression	NN	O	O
did	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
secondary	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
inhibition	NN	O	O
and	NN	O	O
required	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
intracellular	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
RANTES	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
directly	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
efficacy	NN	O	O
of	NN	O	O
these	NN	O	O
agents	NN	O	O
in	NN	O	O
suppressing	NN	O	O
cellular	NN	O	O
infiltration	NN	O	O
and	NN	O	O
to	NN	O	O
their	NN	O	O
anti-inflammatory	NN	O	O
properties	NN	O	O
.	NN	O	O

-DOCSTART-	O

Association	NN	O	O
of	NN	O	O
TRAF1	NN	O	B-protein
,	NN	O	O
TRAF2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TRAF3	NN	O	B-protein
with	NN	O	O
an	NN	O	O
Epstein-Barr	NN	O	B-DNA
virus	NN	O	I-DNA
LMP1	NN	O	I-DNA
domain	NN	O	I-DNA
important	NN	O	O
for	NN	O	O
B-lymphocyte	NN	O	B-cell_type
transformation	NN	O	O
:	NN	O	O
role	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
(	NN	O	I-protein
EBV	NN	O	I-protein
)	NN	O	I-protein
transforming	NN	O	I-protein
protein	NN	O	I-protein
LMP1	NN	O	I-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
constitutively	NN	O	B-protein
activated	NN	O	I-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TNFR	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
an	NN	O	O
intrinsic	NN	O	O
ability	NN	O	O
to	NN	O	O
aggregate	NN	O	O
in	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
and	NN	O	O
an	NN	O	O
association	NN	O	O
of	NN	O	O
its	NN	O	O
cytoplasmic	NN	O	B-protein
carboxyl	NN	O	I-protein
terminus	NN	O	I-protein
(	NN	O	O
CT	NN	O	B-protein
)	NN	O	O
with	NN	O	O
TNFR-associated	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
TRAFs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
EBV-transformed	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
most	NN	O	O
of	NN	O	O
TRAF1	NN	O	B-protein
or	NN	O	O
TRAF3	NN	O	B-protein
and	NN	O	O
5	NN	O	O
%	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
are	NN	O	O
associated	NN	O	O
with	NN	O	O
LMP1	NN	O	B-protein
and	NN	O	O
that	NN	O	O
most	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
is	NN	O	O
associated	NN	O	O
with	NN	O	O
TRAF1	NN	O	B-protein
or	NN	O	O
TRAF3	NN	O	B-protein
.	NN	O	O

TRAF1	NN	O	B-protein
,	NN	O	O
TRAF2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TRAF3	NN	O	B-protein
bind	NN	O	O
to	NN	O	O
a	NN	O	O
single	NN	O	O
site	NN	O	O
in	NN	O	O
the	NN	O	O
LMP1	NN	O	B-protein
CT	NN	O	I-protein
corresponding	NN	O	O
to	NN	O	O
amino	NN	O	O
acids	NN	O	O
(	NN	O	O
aa	NN	O	O
)	NN	O	O
199	NN	O	O
to	NN	O	O
214	NN	O	O
,	NN	O	O
within	NN	O	O
a	NN	O	O
domain	NN	O	O
which	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
B-lymphocyte	NN	O	O
growth	NN	O	O
transformation	NN	O	O
(	NN	O	O
aa	NN	O	O
187	NN	O	O
to	NN	O	O
231	NN	O	O
)	NN	O	O
.	NN	O	O

Further	NN	O	O
deletional	NN	O	O
and	NN	O	O
alanine	NN	O	O
mutagenesis	NN	O	O
analyses	NN	O	O
and	NN	O	O
comparison	NN	O	O
with	NN	O	O
TRAF	NN	O	B-DNA
binding	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
CD40	NN	O	O
,	NN	O	O
in	NN	O	O
CD30	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
LMP1	NN	O	B-protein
of	NN	O	O
other	NN	O	O
lymphycryptoviruses	NN	O	O
provide	NN	O	O
the	NN	O	O
first	NN	O	O
evidence	NN	O	O
that	NN	O	O
PXQXT/S	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
core	NN	O	O
TRAF	NN	O	B-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
negative	NN	O	O
effects	NN	O	O
of	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
LMP1	NN	O	B-DNA
(	NN	O	I-DNA
1-231	NN	O	I-DNA
)	NN	O	I-DNA
core	NN	O	I-DNA
TRAF	NN	O	I-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
on	NN	O	O
TRAF	NN	O	O
binding	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
genetically	NN	O	O
link	NN	O	O
the	NN	O	O
TRAFs	NN	O	B-protein
to	NN	O	O
LMP1	NN	O	B-protein
(	NN	O	O
1-231	NN	O	O
)	NN	O	O
-mediated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
LMP1	NN	O	B-protein
(	NN	O	O
1-231	NN	O	O
)	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
TRAF1	NN	O	B-protein
/TRAF2	NN	O	B-protein
heteroaggregates	NN	O	O
since	NN	O	O
TRAF1	NN	O	B-protein
is	NN	O	O
unique	NN	O	O
among	NN	O	O
the	NN	O	O
TRAFs	NN	O	B-protein
in	NN	O	O
coactivating	NN	O	O
NF-kappaB	NN	O	B-protein
with	NN	O	O
LMP1	NN	O	B-protein
(	NN	O	O
1-231	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
TRAF2	NN	O	B-protein
dominant-negative	NN	O	O
mutant	NN	O	O
can	NN	O	O
block	NN	O	O
LMP1	NN	O	B-protein
(	NN	O	O
1-231	NN	O	O
)	NN	O	O
-mediated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
TRAF1	NN	O	B-protein
coactivation	NN	O	O
,	NN	O	O
and	NN	O	O
30	NN	O	O
%	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
is	NN	O	O
associated	NN	O	O
with	NN	O	O
TRAF1	NN	O	B-protein
in	NN	O	O
EBV-transformed	NN	O	O
B	NN	O	O
cells	NN	O	O
.	NN	O	O

TRAF3	NN	O	B-protein
is	NN	O	O
a	NN	O	O
negative	NN	O	O
modulator	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
(	NN	O	O
1-231	NN	O	O
)	NN	O	O
-mediated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
TRAF1	NN	O	B-protein
,	NN	O	I-protein
-2	NN	O	I-protein
,	NN	O	I-protein
or	NN	O	I-protein
-3	NN	O	I-protein
does	NN	O	O
not	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
terminal	NN	O	B-protein
LMP1	NN	O	I-protein
CT	NN	O	I-protein
aa	NN	O	O
333	NN	O	O
to	NN	O	O
386	NN	O	O
which	NN	O	O
can	NN	O	O
independently	NN	O	O
mediate	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

The	NN	O	O
constitutive	NN	O	O
association	NN	O	O
of	NN	O	O
TRAFs	NN	O	B-protein
with	NN	O	O
LMP1	NN	O	B-protein
through	NN	O	O
the	NN	O	O
aa	NN	O	B-protein
187	NN	O	I-protein
to	NN	O	I-protein
231	NN	O	I-protein
domain	NN	O	I-protein
which	NN	O	O
is	NN	O	O
important	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
primary	NN	O	O
B-lymphocyte	NN	O	B-cell_type
growth	NN	O	O
transformation	NN	O	O
implicates	NN	O	O
TRAF	NN	O	B-protein
aggregation	NN	O	O
in	NN	O	O
LMP1	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lack	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
allergen-specific	NN	O	I-cell_line
Th2	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

IL-12	NN	O	B-protein
is	NN	O	O
a	NN	O	O
powerful	NN	O	O
skewer	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
responses	NN	O	O
toward	NN	O	O
the	NN	O	O
Th1	NN	O	O
phenotype	NN	O	O
by	NN	O	O
inducing	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_line
Th	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
addressed	NN	O	O
the	NN	O	O
question	NN	O	O
of	NN	O	O
whether	NN	O	O
IL-12	NN	O	B-protein
can	NN	O	O
reverse	NN	O	O
established	NN	O	O
Th2	NN	O	O
responses	NN	O	O
into	NN	O	O
Th1/Th0	NN	O	O
responses	NN	O	O
by	NN	O	O
inducing	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
in	NN	O	O
memory	NN	O	B-cell_line
Th2	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
this	NN	O	O
aim	NN	O	O
,	NN	O	O
allergen-specific	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
(	NN	O	O
TCC	NN	O	B-cell_line
)	NN	O	O
were	NN	O	O
generated	NN	O	O
from	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
three	NN	O	O
atopic	NN	O	O
patients	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
cytokine	NN	O	B-protein
profiles	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

The	NN	O	O
majority	NN	O	O
of	NN	O	O
these	NN	O	O
TCC	NN	O	B-cell_line
exhibited	NN	O	O
a	NN	O	O
strongly	NN	O	O
polarized	NN	O	O
Th2	NN	O	O
cytokine	NN	O	B-protein
profile	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
could	NN	O	O
not	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
exogenous	NN	O	O
IL-12	NN	O	B-protein
.	NN	O	O

Only	NN	O	O
those	NN	O	O
TCC	NN	O	B-cell_line
with	NN	O	O
low	NN	O	O
IFN-gamma	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
responded	NN	O	O
to	NN	O	O
IL-12	NN	O	B-protein
by	NN	O	O
additional	NN	O	O
enhancement	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
.	NN	O	O

The	NN	O	O
IL-12	NN	O	B-protein
nonresponsiveness	NN	O	O
of	NN	O	O
the	NN	O	O
Th2	NN	O	B-cell_line
clones	NN	O	I-cell_line
was	NN	O	O
further	NN	O	O
evident	NN	O	O
by	NN	O	O
the	NN	O	O
total	NN	O	O
lack	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
-induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT4	NN	O	B-protein
(	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription-4	NN	O	I-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
is	NN	O	O
typically	NN	O	O
involved	NN	O	O
in	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
IL-12	NN	O	B-protein
also	NN	O	O
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
STAT4-containing	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
these	NN	O	O
Th2	NN	O	B-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

All	NN	O	O
TCC	NN	O	B-cell_line
expressed	NN	O	O
equal	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
low-affinity	NN	O	B-protein
IL-12R	NN	O	I-protein
beta1	NN	O	I-protein
subunit	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
human	NN	O	O
allergen-specific	NN	O	O
Th	NN	O	O
cells	NN	O	O
with	NN	O	O
strongly	NN	O	O
polarized	NN	O	O
Th2	NN	O	O
cytokine	NN	O	B-protein
profiles	NN	O	O
do	NN	O	O
not	NN	O	O
respond	NN	O	O
to	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
produce	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

The	NN	O	O
apparent	NN	O	O
high	NN	O	O
frequency	NN	O	O
of	NN	O	O
IL-12-nonresponsive	NN	O	B-cell_line
Th	NN	O	I-cell_line
cells	NN	O	I-cell_line
within	NN	O	O
the	NN	O	O
allergen-specific	NN	O	O
populations	NN	O	O
in	NN	O	O
atopic	NN	O	O
patients	NN	O	O
predicts	NN	O	O
a	NN	O	O
limited	NN	O	O
skewing	NN	O	O
potential	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
in	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
established	NN	O	O
Th2	NN	O	O
responses	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
affecting	NN	O	O
newly	NN	O	O
recruited	NN	O	O
naive	NN	O	O
Th	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus-encoded	NN	O	I-protein
dUTPase	NN	O	I-protein
:	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
,	NN	O	O
reactivated	NN	O	O
or	NN	O	O
chronic	NN	O	O
virus	NN	O	O
infection	NN	O	O
develop	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
the	NN	O	O
enzyme	NN	O	B-protein
.	NN	O	O

The	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
(	NN	O	I-protein
EBV	NN	O	I-protein
)	NN	O	I-protein
-specific	NN	O	I-protein
dUTPase	NN	O	I-protein
was	NN	O	O
amplified	NN	O	O
from	NN	O	O
virus	NN	O	B-DNA
DNA	NN	O	I-DNA
by	NN	O	O
PCR	NN	O	O
.	NN	O	O

The	NN	O	O
active	NN	O	O
enzyme	NN	O	O
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
and	NN	O	O
in	NN	O	O
insect	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
a	NN	O	O
non-fusion	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
protein	NN	O	O
from	NN	O	O
E.	NN	O	O
coli	NN	O	O
specifically	NN	O	O
converted	NN	O	O
dUTP	NN	O	O
to	NN	O	O
dUMP	NN	O	O
and	NN	O	O
did	NN	O	O
not	NN	O	O
react	NN	O	O
with	NN	O	O
other	NN	O	O
dNTPs	NN	O	O
or	NN	O	O
NTPs	NN	O	O
.	NN	O	O

Preliminary	NN	O	O
experiments	NN	O	O
yielded	NN	O	O
a	NN	O	O
Km	NN	O	O
value	NN	O	O
of	NN	O	O
about	NN	O	O
0.8	NN	O	O
microM	NN	O	O
for	NN	O	O
dUTP	NN	O	O
.	NN	O	O

MAbs	NN	O	O
against	NN	O	O
the	NN	O	O
dUTPase	NN	O	B-protein
reacted	NN	O	O
with	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
approximately	NN	O	O
31	NN	O	O
kDa	NN	O	O
in	NN	O	O
12-O-tetradecanoyl-phorbol-13-acetate	NN	O	B-cell_line
(	NN	O	I-cell_line
TPA	NN	O	I-cell_line
)	NN	O	I-cell_line
-stimulated	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
harbouring	NN	O	O
either	NN	O	O
type	NN	O	O
1	NN	O	O
or	NN	O	O
type	NN	O	O
2	NN	O	O
EBV	NN	O	O
.	NN	O	O

The	NN	O	O
protein	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
untreated	NN	O	B-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
low	NN	O	O
levels	NN	O	O
,	NN	O	O
whereas	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
lytic	NN	O	O
replication	NN	O	O
cycle	NN	O	O
by	NN	O	O
TPA	NN	O	O
treatment	NN	O	O
or	NN	O	O
by	NN	O	O
providing	NN	O	O
the	NN	O	O
immediate	NN	O	B-protein
early	NN	O	I-protein
transactivator	NN	O	I-protein
BZLF1	NN	O	B-protein
in	NN	O	O
trans	NN	O	O
resulted	NN	O	O
in	NN	O	O
increased	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
virus	NN	O	B-protein
dUTPase	NN	O	I-protein
isolated	NN	O	O
from	NN	O	O
EBV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
a	NN	O	O
phosphoprotein	NN	O	B-protein
.	NN	O	O

The	NN	O	O
protein	NN	O	O
expressed	NN	O	O
in	NN	O	O
insect	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
used	NN	O	O
to	NN	O	O
test	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
specific	NN	O	B-protein
antibodies	NN	O	I-protein
in	NN	O	O
sera	NN	O	O
from	NN	O	O
normal	NN	O	O
,	NN	O	O
healthy	NN	O	O
carriers	NN	O	O
and	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
various	NN	O	O
diseases	NN	O	O
.	NN	O	O

While	NN	O	O
the	NN	O	O
sera	NN	O	O
of	NN	O	O
EBV-negative	NN	O	O
individuals	NN	O	O
(	NN	O	O
0/3	NN	O	O
)	NN	O	O
or	NN	O	O
healthy	NN	O	O
carriers	NN	O	O
(	NN	O	O
0/33	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
contain	NN	O	O
detectable	NN	O	O
levels	NN	O	O
of	NN	O	O
antibodies	NN	O	O
,	NN	O	O
patients	NN	O	O
with	NN	O	O
mononucleosis	NN	O	O
(	NN	O	O
5/18	NN	O	O
)	NN	O	O
,	NN	O	O
chronic	NN	O	O
EBV	NN	O	O
infection	NN	O	O
(	NN	O	O
2/7	NN	O	O
)	NN	O	O
,	NN	O	O
EBV	NN	O	O
reactivation	NN	O	O
(	NN	O	O
7/20	NN	O	O
)	NN	O	O
and	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
infection	NN	O	O
(	NN	O	O
5/24	NN	O	O
)	NN	O	O
showed	NN	O	O
elevated	NN	O	O
antibody	NN	O	O
titres	NN	O	O
against	NN	O	O
the	NN	O	O
enzyme	NN	O	O
.	NN	O	O

This	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
dUTPase	NN	O	O
is	NN	O	O
expressed	NN	O	O
during	NN	O	O
EBV	NN	O	O
replication	NN	O	O
and	NN	O	O
reactivation	NN	O	O
.	NN	O	O

The	NN	O	O
enzyme	NN	O	O
might	NN	O	O
therefore	NN	O	O
be	NN	O	O
a	NN	O	O
potential	NN	O	O
target	NN	O	O
for	NN	O	O
drug	NN	O	O
therapy	NN	O	O
under	NN	O	O
conditions	NN	O	O
of	NN	O	O
active	NN	O	O
DNA	NN	O	O
replication	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
catalytic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
pp56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
its	NN	O	O
regulatory	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
is	NN	O	O
sufficient	NN	O	O
for	NN	O	O
inducing	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

The	NN	O	O
lymphoid	NN	O	B-protein
src	NN	O	I-protein
kinase	NN	O	I-protein
pp56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
different	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
responses	NN	O	O
,	NN	O	O
including	NN	O	O
CD4	NN	O	B-protein
-mediated	NN	O	O
enhancement	NN	O	O
of	NN	O	O
Ag-induced	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
early	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
,	NN	O	O
and	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
assumed	NN	O	O
that	NN	O	O
pp56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
acts	NN	O	O
on	NN	O	O
these	NN	O	O
processes	NN	O	O
by	NN	O	O
phosphorylating	NN	O	O
substrates	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
recently	NN	O	O
reported	NN	O	O
that	NN	O	O
the	NN	O	O
NH2	NN	O	B-protein
regulatory	NN	O	B-protein
domain	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
mediate	NN	O	O
CD4	NN	O	B-protein
accessory	NN	O	O
function	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
address	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
the	NN	O	O
regulatory	NN	O	B-protein
and	NN	O	I-protein
catalytic	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
pp56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
to	NN	O	O
another	NN	O	O
function	NN	O	O
of	NN	O	O
this	NN	O	O
enzyme	NN	O	O
independent	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
:	NN	O	O
TCR-induced	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Two	NN	O	O
pp56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
mutants	NN	O	I-protein
lacking	NN	O	O
either	NN	O	O
the	NN	O	O
entire	NN	O	O
catalytic	NN	O	B-protein
domain	NN	O	I-protein
or	NN	O	O
the	NN	O	O
entire	NN	O	O
NH2	NN	O	B-protein
regulatory	NN	O	B-protein
domain	NN	O	I-protein
were	NN	O	O
generated	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
abilities	NN	O	O
to	NN	O	O
trigger	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
TCR-regulated	NN	O	B-DNA
nuclear	NN	O	I-DNA
factor	NN	O	I-DNA
of	NN	O	I-DNA
activated	NN	O	I-DNA
T	NN	O	I-DNA
cells	NN	O	I-DNA
(	NN	O	I-DNA
NF-AT	NN	O	I-DNA
)	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
were	NN	O	O
compared	NN	O	O
.	NN	O	O

Only	NN	O	O
the	NN	O	O
catalytic	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
NH2	NN	O	B-protein
regulatory	NN	O	O
,	NN	O	O
domain	NN	O	O
of	NN	O	O
pp56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
NF-AT	NN	O	B-protein
region	NN	O	O
transactivation	NN	O	O
on	NN	O	O
its	NN	O	O
own	NN	O	O
and	NN	O	O
to	NN	O	O
cooperate	NN	O	O
with	NN	O	O
other	NN	O	O
intracellular	NN	O	O
signals	NN	O	O
to	NN	O	O
trigger	NN	O	O
this	NN	O	O
response	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
pp56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
IL-2	NN	O	B-protein
cytokine	NN	O	B-protein
production	NN	O	O
to	NN	O	O
an	NN	O	O
extent	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
wild-type	NN	O	O
pp56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
different	NN	O	O
domains	NN	O	O
of	NN	O	O
the	NN	O	O
pp56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
molecule	NN	O	O
contribute	NN	O	O
to	NN	O	O
regulate	NN	O	O
distinct	NN	O	O
biologic	NN	O	O
functions	NN	O	O
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
NH2	NN	O	B-protein
regulatory	NN	O	B-protein
domain	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
mediate	NN	O	O
CD4	NN	O	B-protein
accessory	NN	O	O
function	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
pp56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
,	NN	O	O
mimicking	NN	O	O
TCR	NN	O	B-protein
ligation	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
The	NN	O	O
changes	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	O
receptors	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
asthmatic	NN	O	O
subjects	NN	O	O
]	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GCR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
was	NN	O	O
determined	NN	O	O
by	NN	O	O
radioligand-binding	NN	O	O
assay	NN	O	O
in	NN	O	O
extrinsic	NN	O	O
and	NN	O	O
intrinsic	NN	O	O
asthmatics	NN	O	O
.	NN	O	O

Their	NN	O	O
corresponding	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
were	NN	O	O
assessed	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
average	NN	O	O
number	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
in	NN	O	O
asthmatics	NN	O	O
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
linear	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
and	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
asthma	NN	O	O
.	NN	O	O

Besides	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
also	NN	O	O
a	NN	O	O
linear	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
and	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
the	NN	O	O
initial	NN	O	O
attack	NN	O	O
of	NN	O	O
asthma	NN	O	O
.	NN	O	O

No	NN	O	O
difference	NN	O	O
in	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
level	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
asthmatics	NN	O	O
and	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
primary	NN	O	O
and	NN	O	O
general	NN	O	O
impairment	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
metabolism	NN	O	O
in	NN	O	O
the	NN	O	O
asthmatics	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
in	NN	O	O
the	NN	O	O
asthmatics	NN	O	O
is	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

The	NN	O	O
decrease	NN	O	O
of	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
in	NN	O	O
asthmatics	NN	O	O
,	NN	O	O
we	NN	O	O
think	NN	O	O
,	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
heredity	NN	O	O
and	NN	O	O
repeated	NN	O	O
attacks	NN	O	O
of	NN	O	O
asthma	NN	O	O
.	NN	O	O

-DOCSTART-	O

Novel	NN	O	O
aldosterone	NN	O	B-protein
receptors	NN	O	I-protein
:	NN	O	O
specificity-conferring	NN	O	O
mechanism	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
membrane	NN	O	O
.	NN	O	O

Functional	NN	O	O
studies	NN	O	O
in	NN	O	O
extra-renal	NN	O	B-cell_type
,	NN	O	I-cell_type
nonepithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
such	NN	O	O
as	NN	O	O
smooth	NN	O	B-cell_type
muscle	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
more	NN	O	O
recently	NN	O	O
circulating	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
have	NN	O	O
provided	NN	O	O
increasing	NN	O	O
evidence	NN	O	O
that	NN	O	O
aldosterone	NN	O	O
produces	NN	O	O
not	NN	O	O
only	NN	O	O
classical	NN	O	O
genomic	NN	O	O
effects	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
rapid	NN	O	O
non-genomic	NN	O	O
effects	NN	O	O
on	NN	O	O
transmembrane	NN	O	O
electrolyte	NN	O	O
movements	NN	O	O
.	NN	O	O

These	NN	O	O
involve	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
sodium/proton-exchanger	NN	O	B-protein
of	NN	O	O
the	NN	O	O
cell	NN	O	O
membrane	NN	O	O
at	NN	O	O
very	NN	O	O
low	NN	O	O
,	NN	O	O
physiological	NN	O	O
concentrations	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
with	NN	O	O
an	NN	O	O
acute	NN	O	O
onset	NN	O	O
within	NN	O	O
1-2	NN	O	O
minutes	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
messenger	NN	O	O
cascade	NN	O	O
involved	NN	O	O
is	NN	O	O
the	NN	O	O
inositol-1	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
5-trisphosphate/calcium	NN	O	O
pathway	NN	O	O
which	NN	O	O
responds	NN	O	O
over	NN	O	O
the	NN	O	O
same	NN	O	O
rapid	NN	O	O
time	NN	O	O
course	NN	O	O
.	NN	O	O

Such	NN	O	O
changes	NN	O	O
clearly	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
genomic	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
later	NN	O	O
effects	NN	O	O
than	NN	O	O
the	NN	O	O
membrane-related	NN	O	O
rapid	NN	O	O
responses	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
its	NN	O	O
rapid	NN	O	O
time	NN	O	O
course	NN	O	O
the	NN	O	O
unique	NN	O	O
characteristics	NN	O	O
of	NN	O	O
this	NN	O	O
new	NN	O	O
pathway	NN	O	O
for	NN	O	O
steroid	NN	O	O
action	NN	O	O
include	NN	O	O
a	NN	O	O
10000-fold	NN	O	O
selectivity	NN	O	O
for	NN	O	O
aldosterone	NN	O	O
over	NN	O	O
cortisol	NN	O	O
and	NN	O	O
the	NN	O	O
ineffectiveness	NN	O	O
of	NN	O	O
spironolactones	NN	O	O
,	NN	O	O
classical	NN	O	O
mineralocorticoid	NN	O	O
antagonists	NN	O	O
,	NN	O	O
as	NN	O	O
antagonists	NN	O	O
of	NN	O	O
the	NN	O	O
response	NN	O	O
.	NN	O	O

Subsequently	NN	O	O
binding	NN	O	O
sites	NN	O	O
have	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
which	NN	O	O
show	NN	O	O
pharmacological	NN	O	O
(	NN	O	O
aldosterone	NN	O	O
specificity	NN	O	O
)	NN	O	O
and	NN	O	O
kinetic	NN	O	O
(	NN	O	O
high	NN	O	O
turnover	NN	O	O
)	NN	O	O
properties	NN	O	O
identical	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
rapid	NN	O	O
aldosterone	NN	O	O
effects	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
cells	NN	O	O
.	NN	O	O

SDS-PAGE	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
protein	NN	O	I-protein
has	NN	O	O
shown	NN	O	O
a	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
approximately	NN	O	O
50	NN	O	O
kD	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
paper	NN	O	O
reviews	NN	O	O
the	NN	O	O
data	NN	O	O
supporting	NN	O	O
a	NN	O	O
new	NN	O	O
,	NN	O	O
two-step	NN	O	O
model	NN	O	O
for	NN	O	O
non-genomic	NN	O	O
and	NN	O	O
genomic	NN	O	O
aldosterone	NN	O	O
effects	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
suggests	NN	O	O
a	NN	O	O
novel	NN	O	O
specificity-conferring	NN	O	O
mechanism	NN	O	O
for	NN	O	O
mineralocorticoid	NN	O	O
action	NN	O	O
at	NN	O	O
the	NN	O	O
membrane	NN	O	O
level	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
macrophage	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
PU.1	NN	O	B-protein
directs	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
M-CSF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
tissue-specific	NN	O	O
fashion	NN	O	O
from	NN	O	O
two	NN	O	O
distinct	NN	O	O
promoters	NN	O	B-DNA
in	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
and	NN	O	O
the	NN	O	O
placenta	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
further	NN	O	O
understand	NN	O	O
the	NN	O	O
transcription	NN	O	O
factors	NN	O	O
which	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
commitment	NN	O	O
of	NN	O	O
multipotential	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
to	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	O
lineage	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
initiated	NN	O	O
an	NN	O	O
investigation	NN	O	O
of	NN	O	O
the	NN	O	O
factors	NN	O	O
which	NN	O	O
activate	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-protein
receptor	NN	O	I-protein
very	NN	O	O
early	NN	O	O
during	NN	O	O
the	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
process	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
monocytic	NN	O	I-DNA
M-CSF	NN	O	I-DNA
receptor	NN	O	I-DNA
promoter	NN	O	I-DNA
directs	NN	O	O
reporter	NN	O	O
gene	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
tissue-specific	NN	O	O
fashion	NN	O	O
.	NN	O	O

Since	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
few	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
which	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
monocyte	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
the	NN	O	O
macrophage-	NN	O	B-protein
and	NN	O	I-protein
B-cell-specific	NN	O	I-protein
PU.1	NN	O	I-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
PU.1	NN	O	B-protein
binds	NN	O	O
and	NN	O	O
activates	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-DNA
receptor	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
both	NN	O	O
in	NN	O	O
vitro-translated	NN	O	B-protein
PU.1	NN	O	I-protein
and	NN	O	O
PU.1	NN	O	B-protein
from	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
bind	NN	O	O
to	NN	O	O
a	NN	O	O
specific	NN	O	O
site	NN	O	O
in	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-DNA
receptor	NN	O	I-DNA
promoter	NN	O	I-DNA
just	NN	O	O
upstream	NN	O	O
from	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
transcription	NN	O	I-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
this	NN	O	O
site	NN	O	O
which	NN	O	O
eliminate	NN	O	O
PU.1	NN	O	B-protein
binding	NN	O	O
decrease	NN	O	O
M-CSF	NN	O	B-protein
receptor	NN	O	I-protein
promoter	NN	O	O
activity	NN	O	O
significantly	NN	O	O
in	NN	O	O
macrophage	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
only	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
PU.1	NN	O	B-protein
transactivates	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-DNA
receptor	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
nonmacrophage	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
PU.1	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
macrophage	NN	O	O
gene	NN	O	O
regulation	NN	O	O
and	NN	O	O
development	NN	O	O
by	NN	O	O
directing	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
receptor	NN	O	O
for	NN	O	O
a	NN	O	O
key	NN	O	B-protein
macrophage	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Increased	NN	O	O
natural	NN	O	O
killer	NN	O	O
cell	NN	O	O
activity	NN	O	O
correlates	NN	O	O
with	NN	O	O
low	NN	O	O
or	NN	O	O
negative	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
HER-2/neu	NN	O	B-DNA
oncogene	NN	O	I-DNA
in	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
.	NN	O	O

Increased	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
HER-2/neu	NN	O	B-DNA
oncogene	NN	O	I-DNA
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
correlates	NN	O	O
with	NN	O	O
decreased	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
concentration	NN	O	O
and	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
prognostic	NN	O	O
factor	NN	O	O
.	NN	O	O

The	NN	O	O
authors	NN	O	O
investigated	NN	O	O
whether	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
correlation	NN	O	O
between	NN	O	O
HER-2/neu	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
immunologic	NN	O	O
parameters	NN	O	O
representing	NN	O	O
tumor	NN	O	O
defense	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

METHOD	NN	O	O
.	NN	O	O

A	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
investigate	NN	O	O
HER-2/neu	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
chromium-release	NN	O	O
assay	NN	O	O
using	NN	O	O
the	NN	O	O
K562	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
as	NN	O	O
target	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
measure	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
.	NN	O	O

In	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
,	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
compared	NN	O	O
with	NN	O	O
patients	NN	O	O
with	NN	O	O
benign	NN	O	B-cell_type
tumors	NN	O	I-cell_type
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.006	NN	O	O
)	NN	O	O
or	NN	O	O
healthy	NN	O	O
control	NN	O	O
subjects	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.002	NN	O	O
)	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
23.3	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
showed	NN	O	O
an	NN	O	O
overexpression	NN	O	O
of	NN	O	O
HER-2/neu	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Within	NN	O	O
this	NN	O	O
group	NN	O	O
of	NN	O	O
patients	NN	O	O
,	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
(	NN	O	O
45.6	NN	O	O
+/-	NN	O	O
16.1	NN	O	O
%	NN	O	O
)	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
group	NN	O	O
with	NN	O	O
no	NN	O	O
HER-2/neu	NN	O	B-protein
overexpression	NN	O	O
(	NN	O	O
57.3	NN	O	O
+/-	NN	O	O
11.0	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
did	NN	O	O
not	NN	O	O
increase	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
HER-2/neu	NN	O	B-protein
overexpression	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
statistically	NN	O	O
significant	NN	O	O
correlation	NN	O	O
of	NN	O	O
cytolytic	NN	O	O
effector	NN	O	O
cell	NN	O	O
function	NN	O	O
with	NN	O	O
HER-2/neu	NN	O	B-protein
expression	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.003	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
HER-2/neu	NN	O	B-protein
overexpression	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
negative	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
status	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.005	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
add	NN	O	O
further	NN	O	O
evidence	NN	O	O
to	NN	O	O
previous	NN	O	O
observations	NN	O	O
from	NN	O	O
the	NN	O	O
authors	NN	O	O
'	NN	O	O
laboratory	NN	O	O
that	NN	O	O
certain	NN	O	O
tumor	NN	O	O
characteristics	NN	O	O
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
reactions	NN	O	O
of	NN	O	O
the	NN	O	O
host	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
The	NN	O	O
value	NN	O	O
of	NN	O	O
the	NN	O	O
clinical	NN	O	O
test	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
pulmonary	NN	O	O
heart	NN	O	O
disease	NN	O	O
]	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
inquire	NN	O	O
into	NN	O	O
the	NN	O	O
functional	NN	O	O
state	NN	O	O
of	NN	O	O
adrenal	NN	O	O
cortex	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
pulmonary	NN	O	O
heart	NN	O	O
disease	NN	O	O
,	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GCR	NN	O	B-protein
)	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
pulmonary	NN	O	O
heart	NN	O	O
disease	NN	O	O
was	NN	O	O
determined	NN	O	O
with	NN	O	O
radioligand-binding	NN	O	O
assay	NN	O	O
and	NN	O	O
the	NN	O	O
corresponding	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
were	NN	O	O
assessed	NN	O	O
with	NN	O	O
radioimmune	NN	O	O
assays	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
in	NN	O	O
the	NN	O	O
patients	NN	O	O
was	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
and	NN	O	O
it	NN	O	O
was	NN	O	O
increased	NN	O	O
when	NN	O	O
their	NN	O	O
health	NN	O	O
state	NN	O	O
was	NN	O	O
improved	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
still	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
in	NN	O	O
the	NN	O	O
patients	NN	O	O
was	NN	O	O
greatly	NN	O	O
increased	NN	O	O
when	NN	O	O
these	NN	O	O
patients	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
oxygen	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

No	NN	O	O
difference	NN	O	O
in	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
the	NN	O	O
patients	NN	O	O
and	NN	O	O
the	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
(	NN	O	O
P	NN	O	O
>	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
adrenal	NN	O	O
cortex	NN	O	O
may	NN	O	O
be	NN	O	O
improved	NN	O	O
by	NN	O	O
the	NN	O	O
compensation	NN	O	O
mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
lower	NN	O	O
GCR	NN	O	B-protein
number	NN	O	O
was	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
lacking	NN	O	O
of	NN	O	O
oxygen	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
may	NN	O	O
be	NN	O	O
improved	NN	O	O
by	NN	O	O
inhalation	NN	O	O
of	NN	O	O
oxygen	NN	O	O
.	NN	O	O

Therefore	NN	O	O
oxygen	NN	O	O
therapy	NN	O	O
is	NN	O	O
helpful	NN	O	O
in	NN	O	O
raising	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
controlling	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signaling	NN	O	O
via	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
in	NN	O	O
JAK3-deficient	NN	O	B-cell_line
severe	NN	O	I-cell_line
combined	NN	O	I-cell_line
immunodeficiency	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
:	NN	O	O
JAK3	NN	O	B-protein
-dependent	NN	O	O
and	NN	O	O
independent	NN	O	O
pathways	NN	O	O
.	NN	O	O

Both	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
bind	NN	O	O
to	NN	O	O
receptors	NN	O	O
containing	NN	O	O
the	NN	O	O
common	NN	O	B-protein
gamma	NN	O	I-protein
chain	NN	O	I-protein
and	NN	O	O
JAK3	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
JAK3	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
proper	NN	O	O
lymphoid	NN	O	O
development	NN	O	O
,	NN	O	O
the	NN	O	O
precise	NN	O	O
roles	NN	O	O
of	NN	O	O
this	NN	O	O
kinase	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
have	NN	O	O
not	NN	O	O
been	NN	O	O
defined	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
lacking	NN	O	O
JAK3	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
JAK1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT5	NN	O	B-protein
all	NN	O	O
required	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
-mediated	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
JAK1	NN	O	B-protein
,	NN	O	O
STAT6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
insulin	NN	O	B-protein
receptor	NN	O	I-protein
substrates	NN	O	O
1	NN	O	O
and	NN	O	O
2	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
effects	NN	O	O
were	NN	O	O
clearly	NN	O	O
improved	NN	O	O
following	NN	O	O
JAK3	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
signaling	NN	O	O
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
is	NN	O	O
comparatively	NN	O	O
inefficient	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
may	NN	O	O
help	NN	O	O
in	NN	O	O
understanding	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
the	NN	O	O
immunodeficiency	NN	O	O
that	NN	O	O
occurs	NN	O	O
with	NN	O	O
mutations	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
and	NN	O	O
may	NN	O	O
suggest	NN	O	O
a	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
pleiotropic	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Elf-1	NN	O	B-protein
and	NN	O	O
Stat5	NN	O	B-protein
bind	NN	O	O
to	NN	O	O
a	NN	O	O
critical	NN	O	O
element	NN	O	O
in	NN	O	O
a	NN	O	O
new	NN	O	O
enhancer	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin-2	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
Mol	NN	O	O
Cell	NN	O	O
Biol	NN	O	O
1997	NN	O	O
Apr	NN	O	O
;	NN	O	O
17	NN	O	O
(	NN	O	O
4	NN	O	O
)	NN	O	O
:	NN	O	O
2351	NN	O	O
]	NN	O	O

The	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha-chain	NN	O	I-DNA
(	NN	O	I-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
key	NN	O	O
regulator	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
.	NN	O	O

IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
is	NN	O	O
rapidly	NN	O	O
and	NN	O	O
potently	NN	O	O
induced	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogenic	NN	O	O
stimuli	NN	O	O
.	NN	O	O

Interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
stimulates	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

transcription	NN	O	O
,	NN	O	O
thereby	NN	O	O
amplifying	NN	O	O
expression	NN	O	O
of	NN	O	O
its	NN	O	O
own	NN	O	O
high-affinity	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
transcription	NN	O	O
is	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
controlled	NN	O	O
by	NN	O	O
two	NN	O	O
positive	NN	O	B-protein
regulatory	NN	O	I-protein
regions	NN	O	I-protein
,	NN	O	O
PRRI	NN	O	B-DNA
and	NN	O	O
PRRII	NN	O	B-DNA
.	NN	O	O

PRRI	NN	O	B-DNA
is	NN	O	O
an	NN	O	O
inducible	NN	O	B-DNA
proximal	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
located	NN	O	O
between	NN	O	B-DNA
nucleotides	NN	O	I-DNA
-276	NN	O	I-DNA
and	NN	O	I-DNA
-244	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
contains	NN	O	O
NF-kappaB	NN	O	B-DNA
and	NN	O	I-DNA
SRE/CArG	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

PRRII	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
T-cell-specific	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
located	NN	O	O
between	NN	O	O
nucleotides	NN	O	O
-137	NN	O	O
and	NN	O	O
-64	NN	O	O
,	NN	O	O
which	NN	O	O
binds	NN	O	O
the	NN	O	O
T-cell-specific	NN	O	B-protein
Ets	NN	O	I-protein
protein	NN	O	I-protein
Elf-1	NN	O	B-protein
and	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
none	NN	O	O
of	NN	O	O
these	NN	O	O
proximal	NN	O	O
regions	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-2R	NN	O	O
alpha	NN	O	O
transcription	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

To	NN	O	O
find	NN	O	O
new	NN	O	O
regulatory	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
8.5	NN	O	O
kb	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	O
'	NN	O	O
end	NN	O	O
noncoding	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
sequenced	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
an	NN	O	O
86-nucleotide	NN	O	O
fragment	NN	O	O
that	NN	O	O
is	NN	O	O
90	NN	O	O
%	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
recently	NN	O	O
characterized	NN	O	O
murine	NN	O	B-DNA
IL-2-responsive	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
mIL-2rE	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

This	NN	O	O
putative	NN	O	O
human	NN	O	B-DNA
IL-2rE	NN	O	I-DNA
,	NN	O	O
designated	NN	O	O
PRRIII	NN	O	B-DNA
,	NN	O	O
confers	NN	O	O
IL-2	NN	O	B-protein
responsiveness	NN	O	O
on	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

PRRIII	NN	O	B-DNA
contains	NN	O	O
a	NN	O	O
Stat	NN	O	B-DNA
protein	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
that	NN	O	O
overlaps	NN	O	O
with	NN	O	O
an	NN	O	O
EBS	NN	O	B-DNA
motif	NN	O	I-DNA
(	NN	O	O
GASd/EBSd	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

These	NN	O	O
are	NN	O	O
essential	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
inducibility	NN	O	O
of	NN	O	O
PRRIII/CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
.	NN	O	O

IL-2	NN	O	B-protein
induced	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
Stat5a	NN	O	B-protein
and	NN	O	I-protein
b	NN	O	I-protein
proteins	NN	O	I-protein
to	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GASd	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
confirm	NN	O	O
the	NN	O	O
physiological	NN	O	O
relevance	NN	O	O
of	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
we	NN	O	O
carried	NN	O	O
out	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprinting	NN	O	O
experiments	NN	O	O
which	NN	O	O
showed	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
correlated	NN	O	O
with	NN	O	O
occupancy	NN	O	O
of	NN	O	O
the	NN	O	O
GASd	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
GASd/EBSd	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
regulation	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
T-cell-specific	NN	O	B-protein
Elf-1	NN	O	I-protein
factor	NN	O	I-protein
can	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
repressor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Signal	NN	O	O
transduction	NN	O	O
by	NN	O	O
DR3	NN	O	B-protein
,	NN	O	O
a	NN	O	O
death	NN	O	B-protein
domain-containing	NN	O	I-protein
receptor	NN	O	I-protein
related	NN	O	O
to	NN	O	O
TNFR-1	NN	O	B-protein
and	NN	O	O
CD95	NN	O	B-protein
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-1	NN	O	I-protein
(	NN	O	O
TNFR-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
CD95	NN	O	B-protein
(	NN	O	O
also	NN	O	O
called	NN	O	O
Fas	NN	O	B-protein
or	NN	O	O
APO-1	NN	O	B-protein
)	NN	O	O
are	NN	O	O
cytokine	NN	O	B-protein
receptors	NN	O	I-protein
that	NN	O	O
engage	NN	O	O
the	NN	O	O
apoptosis	NN	O	O
pathway	NN	O	O
through	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
intracellular	NN	O	O
homology	NN	O	O
,	NN	O	O
designated	NN	O	O
the	NN	O	O
``	NN	O	O
death	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O
''	NN	O	O
Another	NN	O	O
death	NN	O	B-protein
domain-containing	NN	O	I-protein
member	NN	O	I-protein
of	NN	O	O
the	NN	O	O
TNFR	NN	O	B-protein
family	NN	O	I-protein
,	NN	O	O
death	NN	O	B-protein
receptor	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
DR3	NN	O	B-protein
)	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
and	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
induce	NN	O	O
both	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
DR3	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
restricted	NN	O	O
to	NN	O	O
tissues	NN	O	O
enriched	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

DR3	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
complex	NN	O	O
of	NN	O	O
intracellular	NN	O	B-protein
signaling	NN	O	I-protein
molecules	NN	O	I-protein
including	NN	O	O
TRADD	NN	O	B-protein
,	NN	O	O
TRAF2	NN	O	B-protein
,	NN	O	O
FADD	NN	O	B-protein
,	NN	O	O
and	NN	O	O
FLICE	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
DR3	NN	O	B-protein
likely	NN	O	O
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
lymphocyte	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p52	NN	O	B-protein
,	NN	O	O
and	NN	O	O
RelB	NN	O	B-protein
proteins	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
accessory	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
in	NN	O	O
situ	NN	O	O
.	NN	O	O

The	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p52	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
and	NN	O	O
RelB	NN	O	B-protein
,	NN	O	O
constitute	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
positive	NN	O	O
regulation	NN	O	O
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
genes	NN	O	O
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
that	NN	O	O
RelB	NN	O	B-protein
knockout	NN	O	O
mice	NN	O	O
have	NN	O	O
no	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
DC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

An	NN	O	O
overexpression	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
has	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
follicular	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
FDC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

A	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
were	NN	O	O
key	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
functions	NN	O	O
of	NN	O	O
accessory	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
in	NN	O	O
situ	NN	O	O
the	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
accessory	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
by	NN	O	O
immunohistochemistry	NN	O	O
and	NN	O	O
double	NN	O	O
labeling	NN	O	O
by	NN	O	O
immunofluorescence	NN	O	O
from	NN	O	O
five	NN	O	O
normal	NN	O	O
human	NN	O	O
tonsils	NN	O	O
and	NN	O	O
five	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
with	NN	O	O
follicular	NN	O	O
hyperplasia	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
proteins	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
all	NN	O	O
cell	NN	O	O
types	NN	O	O
including	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
germinal	NN	O	O
centers	NN	O	O
GC	NN	O	O
,	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p52	NN	O	B-protein
,	NN	O	O
and	NN	O	O
RelB	NN	O	B-protein
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
FDC	NN	O	B-cell_type
only	NN	O	O
and	NN	O	O
were	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
CD68+	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
T	NN	O	O
cell	NN	O	O
areas	NN	O	O
,	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p52	NN	O	B-protein
,	NN	O	O
and	NN	O	O
RelB	NN	O	B-protein
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
HLA-DR+	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
an	NN	O	O
antigen-presenting	NN	O	O
cell	NN	O	O
(	NN	O	O
APC	NN	O	O
)	NN	O	O
morphology	NN	O	O
.	NN	O	O

p52	NN	O	B-protein
and	NN	O	O
RelB	NN	O	B-protein
were	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
in	NN	O	O
both	NN	O	O
CD1a+	NN	O	B-cell_line
and	NN	O	O
CD68+	NN	O	B-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
area	NN	O	O
,	NN	O	O
whereas	NN	O	O
p50	NN	O	B-protein
was	NN	O	O
found	NN	O	O
only	NN	O	O
in	NN	O	O
CD68-	NN	O	B-cell_line
and	NN	O	I-cell_line
CD1a-	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Cells	NN	O	O
with	NN	O	O
nuclear	NN	O	O
p50	NN	O	B-protein
were	NN	O	O
negative	NN	O	O
for	NN	O	O
the	NN	O	O
CD38	NN	O	B-protein
,	NN	O	O
CD20	NN	O	B-protein
and	NN	O	O
CD2	NN	O	B-protein
markers	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
,	NN	O	O
physiologically	NN	O	O
,	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
nuclear	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p52	NN	O	B-protein
and	NN	O	O
RelB	NN	O	B-protein
are	NN	O	O
restricted	NN	O	O
to	NN	O	O
accessory	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
,	NN	O	O
which	NN	O	O
include	NN	O	O
FDC	NN	O	B-cell_type
in	NN	O	O
GC	NN	O	O
,	NN	O	O
and	NN	O	O
DC	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	O
in	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
zone	NN	O	O
,	NN	O	O
that	NN	O	O
specialized	NN	O	O
scavenger	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
from	NN	O	O
GC	NN	O	B-cell_type
do	NN	O	O
not	NN	O	O
have	NN	O	O
detectable	NN	O	O
levels	NN	O	O
of	NN	O	O
p52	NN	O	B-protein
and	NN	O	O
RelB	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
macrophages	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
area	NN	O	O
,	NN	O	O
known	NN	O	O
to	NN	O	O
present	NN	O	O
the	NN	O	O
antigen	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
do	NN	O	O
have	NN	O	O
both	NN	O	O
nuclear	NN	O	O
p52	NN	O	B-protein
and	NN	O	O
RelB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
zone	NN	O	O
,	NN	O	O
p52	NN	O	B-protein
and	NN	O	O
RelB	NN	O	B-protein
are	NN	O	O
located	NN	O	O
in	NN	O	O
nuclei	NN	O	O
of	NN	O	O
both	NN	O	O
CD1a+	NN	O	B-cell_line
,	NN	O	O
CD68+	NN	O	B-cell_line
or	NN	O	O
both	NN	O	O
,	NN	O	O
cells	NN	O	O
APC	NN	O	B-cell_type
,	NN	O	O
whereas	NN	O	O
p50	NN	O	B-protein
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
CD1a-	NN	O	B-cell_type
and	NN	O	O
CD68-	NN	O	B-cell_type
APC	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
different	NN	O	O
patterns	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p52	NN	O	B-protein
and	NN	O	O
RelB	NN	O	B-protein
protein	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
may	NN	O	O
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
their	NN	O	O
different	NN	O	O
roles	NN	O	O
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tyrosines	NN	O	O
113	NN	O	O
,	NN	O	O
128	NN	O	O
,	NN	O	O
and	NN	O	O
145	NN	O	O
of	NN	O	O
SLP-76	NN	O	B-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
augmentation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

SLP-76	NN	O	B-protein
(	NN	O	O
SH2	NN	O	B-protein
domain	NN	O	I-protein
leukocyte	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
76	NN	O	O
kDa	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
recently	NN	O	O
identified	NN	O	O
substrate	NN	O	O
of	NN	O	O
the	NN	O	O
TCR-stimulated	NN	O	B-protein
protein	NN	O	I-protein
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
that	NN	O	O
functions	NN	O	O
in	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
cascade	NN	O	O
linking	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
with	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
engagement	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
results	NN	O	O
in	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
SLP-76	NN	O	B-protein
in	NN	O	O
its	NN	O	O
amino-terminal	NN	O	B-protein
acidic	NN	O	I-protein
region	NN	O	I-protein
.	NN	O	O

Two	NN	O	O
tyrosines	NN	O	O
(	NN	O	O
Y113	NN	O	O
and	NN	O	O
Y128	NN	O	O
)	NN	O	O
fall	NN	O	O
within	NN	O	O
an	NN	O	O
identical	NN	O	O
five	NN	O	O
amino-acid	NN	O	O
motif	NN	O	O
and	NN	O	O
are	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
phosphorylated	NN	O	O
upon	NN	O	O
TCR	NN	O	B-protein
ligation	NN	O	O
.	NN	O	O

Although	NN	O	O
mutation	NN	O	O
of	NN	O	O
either	NN	O	O
Y113	NN	O	O
and	NN	O	O
Y128	NN	O	O
has	NN	O	O
a	NN	O	O
minimal	NN	O	O
effect	NN	O	O
on	NN	O	O
SLP-76	NN	O	B-protein
function	NN	O	O
,	NN	O	O
mutation	NN	O	O
of	NN	O	O
both	NN	O	O
residues	NN	O	O
decreases	NN	O	O
significantly	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
SLP-76	NN	O	B-protein
to	NN	O	O
promote	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

A	NN	O	O
third	NN	O	O
tyrosine	NN	O	O
within	NN	O	O
the	NN	O	O
amino-terminal	NN	O	B-protein
region	NN	O	I-protein
(	NN	O	O
Y145	NN	O	O
)	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
most	NN	O	O
important	NN	O	O
for	NN	O	O
optimal	NN	O	O
SLP-76	NN	O	B-protein
function	NN	O	O
,	NN	O	O
as	NN	O	O
altering	NN	O	O
it	NN	O	O
alone	NN	O	O
to	NN	O	O
phenylalanine	NN	O	O
has	NN	O	O
a	NN	O	O
potent	NN	O	O
impact	NN	O	O
on	NN	O	O
SLP-76	NN	O	B-protein
augmentation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-DNA
interferon	NN	O	I-DNA
regulatory	NN	O	I-DNA
factor	NN	O	I-DNA
2	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Intron-exon	NN	O	O
organization	NN	O	O
and	NN	O	O
functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

Interferon	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	O
IRF-2	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
protein	NN	O	I-protein
that	NN	O	O
terminates	NN	O	O
interferon	NN	O	B-protein
beta	NN	O	I-protein
expression	NN	O	O
initiated	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
the	NN	O	O
genomic	NN	O	B-DNA
DNA	NN	O	I-DNA
for	NN	O	O
human	NN	O	B-DNA
IRF-2	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
determined	NN	O	O
the	NN	O	O
intron-exon	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IRF-2	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
mapped	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
transcription	NN	O	I-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
identified	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
potential	NN	O	O
regulatory	NN	O	O
elements	NN	O	O
in	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
localized	NN	O	O
the	NN	O	O
IRF-2	NN	O	B-DNA
gene	NN	O	I-DNA
on	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
4	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
IRF-2	NN	O	B-protein
promoter	NN	O	B-DNA
region	NN	O	I-DNA
contains	NN	O	O
a	NN	O	O
CpG	NN	O	O
island	NN	O	O
,	NN	O	O
with	NN	O	O
several	NN	O	O
GC	NN	O	B-DNA
boxes	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
putative	NN	O	O
NF-kappa	NN	O	B-DNA
B-binding	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
CAAT	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
no	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
was	NN	O	O
linked	NN	O	O
with	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
is	NN	O	O
inducible	NN	O	O
by	NN	O	O
both	NN	O	O
interferons	NN	O	B-protein
(	NN	O	O
interferon-alpha	NN	O	B-protein
and	NN	O	I-protein
-gamma	NN	O	I-protein
)	NN	O	O
and	NN	O	O
interferon	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
.	NN	O	O

The	NN	O	O
region	NN	O	O
which	NN	O	O
induced	NN	O	O
these	NN	O	O
inductions	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
being	NN	O	O
confined	NN	O	O
to	NN	O	O
40	NN	O	B-DNA
nucleotides	NN	O	I-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
transcriptional	NN	O	I-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
by	NN	O	O
testing	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
clones	NN	O	O
with	NN	O	O
truncated	NN	O	B-DNA
promoter	NN	O	I-DNA
of	NN	O	O
IRF-2	NN	O	B-protein
.	NN	O	O

This	NN	O	O
region	NN	O	O
contains	NN	O	O
elements	NN	O	O
which	NN	O	O
are	NN	O	O
shared	NN	O	O
with	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
enhancers	NN	O	I-DNA
of	NN	O	O
other	NN	O	O
genes	NN	O	O
including	NN	O	O
interferon	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
interferon	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
and	NN	O	O
interferon-inducible	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
interferon	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
not	NN	O	O
only	NN	O	O
triggers	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
interferon	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
limiting	NN	O	O
the	NN	O	O
duration	NN	O	O
of	NN	O	O
this	NN	O	O
response	NN	O	O
by	NN	O	O
activating	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
IRF-2	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Interferon	NN	O	B-protein
alpha	NN	O	I-protein
selectively	NN	O	O
affects	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
myeloid	NN	O	I-protein
cell	NN	O	I-protein
nuclear	NN	O	I-protein
differentiation	NN	O	I-protein
antigen	NN	O	I-protein
in	NN	O	O
late	NN	O	B-cell_type
stage	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_type
but	NN	O	O
not	NN	O	O
the	NN	O	O
granulocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
human	NN	O	B-protein
myeloid	NN	O	I-protein
cell	NN	O	I-protein
nuclear	NN	O	I-protein
differentiation	NN	O	I-protein
antigen	NN	O	I-protein
(	NN	O	O
MNDA	NN	O	B-protein
)	NN	O	O
is	NN	O	O
expressed	NN	O	O
constitutively	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
myeloid	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
appearing	NN	O	O
in	NN	O	O
myeloblast	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
some	NN	O	O
cases	NN	O	O
of	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
and	NN	O	O
consistently	NN	O	O
being	NN	O	O
detected	NN	O	O
in	NN	O	O
promyelocyte	NN	O	B-cell_type
stage	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
all	NN	O	O
later	NN	O	B-cell_type
stage	NN	O	I-cell_type
cells	NN	O	I-cell_type
including	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
human	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
HL-60	NN	O	B-cell_line
,	NN	O	O
U937	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
THP-1	NN	O	B-cell_line
,	NN	O	O
express	NN	O	O
similar	NN	O	O
levels	NN	O	O
of	NN	O	O
immunochemically	NN	O	O
detectable	NN	O	O
MNDA	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
,	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
MNDA	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
primary	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
is	NN	O	O
very	NN	O	O
low	NN	O	O
it	NN	O	O
was	NN	O	O
up-regulated	NN	O	O
at	NN	O	O
6	NN	O	O
h	NN	O	O
following	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
alpha	NN	O	I-protein
on	NN	O	O
the	NN	O	O
MNDA	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
HL-60	NN	O	B-cell_line
,	NN	O	O
U937	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
THP-1	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
MNDA	NN	O	B-RNA
mRNA	NN	O	I-RNA
level	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
other	NN	O	O
agents	NN	O	O
including	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
,	NN	O	O
endotoxin	NN	O	B-protein
,	NN	O	O
poly	NN	O	B-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
.poly	NN	O	I-protein
(	NN	O	I-protein
C	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
and	NN	O	O
FMLP	NN	O	B-protein
.	NN	O	O

The	NN	O	O
MNDA	NN	O	B-RNA
mRNA	NN	O	I-RNA
level	NN	O	O
in	NN	O	O
the	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
was	NN	O	O
also	NN	O	O
unaffected	NN	O	O
by	NN	O	O
the	NN	O	O
latter	NN	O	O
four	NN	O	O
agents	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
differentiation	NN	O	O
in	NN	O	O
the	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
induces	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
which	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
MNDA	NN	O	B-RNA
mRNA	NN	O	I-RNA
level	NN	O	O
.	NN	O	O

This	NN	O	O
reduced	NN	O	O
level	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
could	NN	O	O
then	NN	O	O
be	NN	O	O
elevated	NN	O	O
with	NN	O	O
subsequent	NN	O	O
interferon	NN	O	B-protein
alpha	NN	O	I-protein
treatment	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
on	NN	O	O
MNDA	NN	O	B-RNA
mRNA	NN	O	I-RNA
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
since	NN	O	O
inhibiting	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
MNDA	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
variation	NN	O	O
in	NN	O	O
MNDA	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
were	NN	O	O
not	NN	O	O
observed	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
alpha	NN	O	I-protein
to	NN	O	O
up-regulate	NN	O	O
MNDA	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
treated	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
observations	NN	O	O
made	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
YB-1	NN	O	B-protein
in	NN	O	O
human	NN	O	O
T-cell	NN	O	O
lymphotropic	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
basal	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Sequences	NN	O	O
which	NN	O	O
control	NN	O	O
basal	NN	O	O
human	NN	O	O
T-cell	NN	O	O
lymphotropic	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
transcription	NN	O	O
likely	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
initiation	NN	O	O
and	NN	O	O
maintenance	NN	O	O
of	NN	O	O
virus	NN	O	O
replication	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
identified	NN	O	O
and	NN	O	O
analyzed	NN	O	O
a	NN	O	O
45-nucleotide	NN	O	B-DNA
sequence	NN	O	I-DNA
(	NN	O	O
downstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
1	NN	O	I-DNA
[	NN	O	O
DRE	NN	O	B-DNA
1	NN	O	I-DNA
]	NN	O	O
)	NN	O	O
,	NN	O	O
+195	NN	O	B-DNA
to	NN	O	I-DNA
+240	NN	O	I-DNA
,	NN	O	O
at	NN	O	O
the	NN	O	O
boundary	NN	O	O
of	NN	O	O
the	NN	O	O
R/U5	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
which	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
HTLV-I	NN	O	O
basal	NN	O	O
transcription	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
protein	NN	O	O
,	NN	O	O
p37	NN	O	O
,	NN	O	O
which	NN	O	O
specifically	NN	O	O
bound	NN	O	O
to	NN	O	O
DRE	NN	O	B-DNA
1	NN	O	I-DNA
.	NN	O	O

An	NN	O	O
affinity	NN	O	O
column	NN	O	O
fraction	NN	O	O
,	NN	O	O
containing	NN	O	O
p37	NN	O	B-protein
,	NN	O	O
stimulated	NN	O	O
HTLV-I	NN	O	O
transcription	NN	O	O
approximately	NN	O	O
12-fold	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
(	NN	O	O
15B-7	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
from	NN	O	O
a	NN	O	O
Jurkat	NN	O	B-DNA
expression	NN	O	I-DNA
library	NN	O	I-DNA
,	NN	O	O
that	NN	O	O
binds	NN	O	O
specifically	NN	O	O
to	NN	O	O
the	NN	O	O
DRE	NN	O	B-DNA
1	NN	O	I-DNA
regulatory	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
the	NN	O	O
cDNA	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
similarly	NN	O	O
to	NN	O	O
the	NN	O	O
results	NN	O	O
obtained	NN	O	O
with	NN	O	O
purified	NN	O	O
Jurkat	NN	O	O
protein	NN	O	O
,	NN	O	O
was	NN	O	O
decreased	NN	O	O
by	NN	O	O
introduction	NN	O	O
of	NN	O	O
site-specific	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
DRE	NN	O	B-DNA
1	NN	O	I-DNA
regulatory	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
and	NN	O	O
translation	NN	O	O
of	NN	O	O
15B-7	NN	O	B-DNA
cDNA	NN	O	I-DNA
produced	NN	O	O
a	NN	O	O
fusion	NN	O	B-protein
protein	NN	O	I-protein
which	NN	O	O
bound	NN	O	O
specifically	NN	O	O
to	NN	O	O
the	NN	O	O
HTLV-I	NN	O	O
+195	NN	O	O
to	NN	O	O
+240	NN	O	O
oligonucleotide	NN	O	O
.	NN	O	O

The	NN	O	O
partial	NN	O	B-DNA
cDNA	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
protein	NN	O	O
which	NN	O	O
is	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
196	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
of	NN	O	O
the	NN	O	O
36-kDa	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
YB-1	NN	O	B-protein
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
a	NN	O	O
YB-1	NN	O	B-DNA
expression	NN	O	I-DNA
plasmid	NN	O	I-DNA
increases	NN	O	O
HTLV-I	NN	O	O
basal	NN	O	O
transcription	NN	O	O
approximately	NN	O	O
14-fold	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
weight	NN	O	O
,	NN	O	O
DNA-binding	NN	O	O
characteristics	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
transactivation	NN	O	O
activity	NN	O	O
,	NN	O	O
we	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
previously	NN	O	O
identified	NN	O	O
DRE	NN	O	B-DNA
1	NN	O	I-DNA
-binding	NN	O	O
protein	NN	O	O
,	NN	O	O
p37	NN	O	B-protein
,	NN	O	O
is	NN	O	O
YB-1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Changes	NN	O	O
in	NN	O	O
triiodothyronine	NN	O	O
(	NN	O	O
T3	NN	O	O
)	NN	O	O
mononuclear	NN	O	O
leukocyte	NN	O	O
receptor	NN	O	O
kinetics	NN	O	O
after	NN	O	O
T3	NN	O	O
administration	NN	O	O
and	NN	O	O
multiple	NN	O	O
cold-air	NN	O	O
exposures	NN	O	O
.	NN	O	O

Repeated	NN	O	O
cold-air	NN	O	O
exposures	NN	O	O
increase	NN	O	O
human	NN	O	O
triiodothyronine	NN	O	O
(	NN	O	O
T3	NN	O	O
)	NN	O	O
plasma	NN	O	O
clearance	NN	O	O
rates	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
T3	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
NT3R	NN	O	B-protein
)	NN	O	O
in	NN	O	O
this	NN	O	O
condition	NN	O	O
,	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
were	NN	O	O
analyzed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
MNL	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
supplemented	NN	O	O
one	NN	O	O
group	NN	O	O
of	NN	O	O
individuals	NN	O	O
with	NN	O	O
a	NN	O	O
daily	NN	O	O
oral	NN	O	O
replacement	NN	O	O
dose	NN	O	O
of	NN	O	O
T3	NN	O	O
to	NN	O	O
isolate	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
serum	NN	O	O
thyroxine	NN	O	O
(	NN	O	O
T4	NN	O	O
)	NN	O	O
and	NN	O	O
thyrotropin	NN	O	O
(	NN	O	O
TSH	NN	O	O
)	NN	O	O
levels	NN	O	O
on	NN	O	O
receptor	NN	O	O
kinetics	NN	O	O
.	NN	O	O

The	NN	O	O
subjects	NN	O	O
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
cold	NN	O	O
air	NN	O	O
(	NN	O	O
4	NN	O	O
degrees	NN	O	O
C	NN	O	O
)	NN	O	O
twice/d	NN	O	O
,	NN	O	O
30	NN	O	O
min/exposure	NN	O	O
,	NN	O	O
for	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
80	NN	O	O
exposures	NN	O	O
.	NN	O	O

The	NN	O	O
T3-subjects	NN	O	O
received	NN	O	O
placebo	NN	O	O
[	NN	O	O
n	NN	O	O
=	NN	O	O
8	NN	O	O
]	NN	O	O
and	NN	O	O
the	NN	O	O
T3	NN	O	O
+	NN	O	O
subjects	NN	O	O
received	NN	O	O
T3	NN	O	O
(	NN	O	O
30	NN	O	O
micrograms/d	NN	O	O
)	NN	O	O
[	NN	O	O
n	NN	O	O
=	NN	O	O
8	NN	O	O
]	NN	O	O
in	NN	O	O
a	NN	O	O
double-blind	NN	O	O
fashion	NN	O	O
.	NN	O	O

Mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
were	NN	O	O
isolated	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
before	NN	O	O
the	NN	O	O
cold	NN	O	O
exposure	NN	O	O
and	NN	O	O
drug	NN	O	O
regimen	NN	O	O
began	NN	O	O
,	NN	O	O
and	NN	O	O
then	NN	O	O
after	NN	O	O
every	NN	O	O
20	NN	O	O
exposures	NN	O	O
.	NN	O	O

The	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
and	NN	O	O
maximum	NN	O	O
binding	NN	O	O
capacity	NN	O	O
(	NN	O	O
MBC	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
NT3R	NN	O	B-protein
values	NN	O	O
were	NN	O	O
log	NN	O	O
transformed	NN	O	O
to	NN	O	O
minimize	NN	O	O
between-subject	NN	O	O
variability	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
T3+	NN	O	O
group	NN	O	O
,	NN	O	O
serum	NN	O	O
total	NN	O	O
thyroxine	NN	O	O
(	NN	O	O
TT4	NN	O	O
)	NN	O	O
,	NN	O	O
free	NN	O	O
T4	NN	O	O
(	NN	O	O
FT4	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
TSH	NN	O	O
were	NN	O	O
approx	NN	O	O
50	NN	O	O
%	NN	O	O
lower	NN	O	O
than	NN	O	O
both	NN	O	O
basal	NN	O	O
and	NN	O	O
T3-values	NN	O	O
.	NN	O	O

The	NN	O	O
log10Kd	NN	O	O
increased	NN	O	O
0.304	NN	O	O
+/-	NN	O	O
0.139	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.04	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
log10MBC	NN	O	O
increased	NN	O	O
0.49	NN	O	O
+/-	NN	O	O
0.10	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
T3+	NN	O	O
subjects	NN	O	O
compared	NN	O	O
to	NN	O	O
baseline	NN	O	O
.	NN	O	O

This	NN	O	O
change	NN	O	O
in	NN	O	O
MBC	NN	O	O
represents	NN	O	O
a	NN	O	O
311	NN	O	O
%	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
MBC	NN	O	O
over	NN	O	O
baseline	NN	O	O
and	NN	O	O
a	NN	O	O
fivefold	NN	O	O
increase	NN	O	O
over	NN	O	O
placebo-treated	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
T3	NN	O	O
-group	NN	O	O
showed	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
MBC	NN	O	O
over	NN	O	O
the	NN	O	O
study	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
describe	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
the	NN	O	O
rapid	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
NT3R	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
combined	NN	O	O
influence	NN	O	O
of	NN	O	O
cold	NN	O	O
exposure	NN	O	O
and	NN	O	O
reduced	NN	O	O
circulating	NN	O	O
T4	NN	O	O
and	NN	O	O
TSH	NN	O	O
.	NN	O	O

-DOCSTART-	O

HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
CD40-activated	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

CD40-stimulated	NN	O	B-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
are	NN	O	O
highly	NN	O	O
permissive	NN	O	O
to	NN	O	O
a	NN	O	O
productive	NN	O	O
infection	NN	O	O
by	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
.	NN	O	O

In	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
contains	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
control	NN	O	I-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
virus	NN	O	O
,	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
assays	NN	O	O
with	NN	O	O
plasmids	NN	O	O
containing	NN	O	O
deleted	NN	O	O
parts	NN	O	O
of	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
region	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
a	NN	O	O
reporter	NN	O	O
gene	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
essential	NN	O	O
for	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
CD40-stimulated	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
and	NN	O	O
supershift	NN	O	O
assays	NN	O	O
revealed	NN	O	O
that	NN	O	O
important	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
composed	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
c-Rel	NN	O	B-protein
NF-kappaB	NN	O	B-protein
subunits	NN	O	I-protein
was	NN	O	O
present	NN	O	O
in	NN	O	O
nuclei	NN	O	O
of	NN	O	O
CD40-stimulated	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
confirm	NN	O	O
at	NN	O	O
a	NN	O	O
molecular	NN	O	O
level	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
to	NN	O	O
replicate	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
this	NN	O	O
activity	NN	O	O
is	NN	O	O
strongly	NN	O	O
associated	NN	O	O
with	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Sequence	NN	O	O
analysis	NN	O	O
and	NN	O	O
expression	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
human	NN	O	O
FOSB	NN	O	B-DNA
gene	NN	O	I-DNA
(	NN	O	O
G0S3	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

G0S3	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
putative	NN	O	B-DNA
G0/G1	NN	O	I-DNA
switch	NN	O	I-DNA
regulatory	NN	O	I-DNA
genes	NN	O	I-DNA
(	NN	O	O
G0S	NN	O	B-DNA
genes	NN	O	I-DNA
)	NN	O	O
selected	NN	O	O
by	NN	O	O
screening	NN	O	O
cDNA	NN	O	B-DNA
libraries	NN	O	I-DNA
prepared	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
cultured	NN	O	O
for	NN	O	O
2	NN	O	O
hr	NN	O	O
with	NN	O	O
lectin	NN	O	B-protein
and	NN	O	O
cycloheximide	NN	O	O
.	NN	O	O

The	NN	O	O
sequence	NN	O	O
shows	NN	O	O
high	NN	O	O
homology	NN	O	O
with	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
FOSB	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
AP1	NN	O	B-protein
transcriptional	NN	O	I-protein
regulator	NN	O	I-protein
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
cDNA	NN	O	B-DNA
and	NN	O	O
genomic	NN	O	O
sequences	NN	O	O
reveals	NN	O	O
a	NN	O	O
4-exon	NN	O	B-DNA
structure	NN	O	I-DNA
characteristic	NN	O	O
of	NN	O	O
the	NN	O	O
FOS	NN	O	B-DNA
family	NN	O	I-DNA
of	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Freshly	NN	O	O
isolated	NN	O	O
cells	NN	O	O
show	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
FOSB/G0S3	NN	O	B-RNA
and	NN	O	I-RNA
FOS/G0S7	NN	O	I-RNA
mRNAs	NN	O	I-RNA
,	NN	O	O
which	NN	O	O
decline	NN	O	O
rapidly	NN	O	O
during	NN	O	O
incubation	NN	O	O
in	NN	O	O
culture	NN	O	O
medium	NN	O	O
.	NN	O	O

The	NN	O	O
kinetics	NN	O	O
of	NN	O	O
expression	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
high	NN	O	O
initial	NN	O	O
levels	NN	O	O
are	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
isolation	NN	O	O
procedure	NN	O	O
,	NN	O	O
and	NN	O	O
do	NN	O	O
not	NN	O	O
reflect	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
cells	NN	O	O
preincubated	NN	O	O
for	NN	O	O
a	NN	O	O
day	NN	O	O
,	NN	O	O
levels	NN	O	O
of	NN	O	O
FOS	NN	O	B-RNA
mRNA	NN	O	I-RNA
reach	NN	O	O
a	NN	O	O
maximum	NN	O	O
20	NN	O	O
min	NN	O	O
after	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
lectin	NN	O	B-protein
and	NN	O	O
decline	NN	O	O
to	NN	O	O
control	NN	O	O
levels	NN	O	O
over	NN	O	O
the	NN	O	O
next	NN	O	O
3	NN	O	O
hr	NN	O	O
.	NN	O	O

Levels	NN	O	O
of	NN	O	O
FOSB	NN	O	B-RNA
mRNA	NN	O	I-RNA
reach	NN	O	O
a	NN	O	O
maximum	NN	O	O
40	NN	O	O
min	NN	O	O
after	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
lectin	NN	O	B-protein
and	NN	O	O
decline	NN	O	O
to	NN	O	O
control	NN	O	O
levels	NN	O	O
over	NN	O	O
the	NN	O	O
next	NN	O	O
6	NN	O	O
hr	NN	O	O
.	NN	O	O

In	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
cells	NN	O	O
,	NN	O	O
both	NN	O	O
FOS	NN	O	B-RNA
and	NN	O	I-RNA
FOSB	NN	O	I-RNA
mRNAs	NN	O	I-RNA
increase	NN	O	O
dramatically	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
inhibitor	NN	O	O
cycloheximide	NN	O	O
.	NN	O	O

In	NN	O	O
preincubated	NN	O	O
cells	NN	O	O
,	NN	O	O
the	NN	O	O
cycloheximide	NN	O	O
response	NN	O	O
is	NN	O	O
decreased	NN	O	O
,	NN	O	O
especially	NN	O	O
in	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
FOSB	NN	O	B-DNA
.	NN	O	O

These	NN	O	O
differences	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
FOS	NN	O	B-DNA
and	NN	O	O
FOSB	NN	O	B-DNA
suggest	NN	O	O
different	NN	O	O
roles	NN	O	O
and	NN	O	O
regulation	NN	O	O
.	NN	O	O

Regions	NN	O	O
of	NN	O	O
low	NN	O	O
base	NN	O	O
order-dependent	NN	O	O
stem-loop	NN	O	O
potential	NN	O	O
in	NN	O	O
the	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
are	NN	O	O
defined	NN	O	O
.	NN	O	O

These	NN	O	O
indicate	NN	O	O
where	NN	O	O
base	NN	O	O
order	NN	O	O
has	NN	O	O
been	NN	O	O
adapted	NN	O	O
for	NN	O	O
purposes	NN	O	O
other	NN	O	O
than	NN	O	O
stem-loop	NN	O	O
stability	NN	O	O
(	NN	O	O
e.g.	NN	O	O
,	NN	O	O
encoding	NN	O	O
proteins	NN	O	O
or	NN	O	O
gene	NN	O	O
regulation	NN	O	O
)	NN	O	O
.	NN	O	O

Regions	NN	O	O
of	NN	O	O
low	NN	O	O
potential	NN	O	O
in	NN	O	O
a	NN	O	O
68.5-kb	NN	O	O
genomic	NN	O	O
segment	NN	O	O
containing	NN	O	O
the	NN	O	O
FOSB	NN	O	B-DNA
gene	NN	O	I-DNA
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
potential	NN	O	O
may	NN	O	O
help	NN	O	O
locate	NN	O	O
genes	NN	O	O
in	NN	O	O
uncharted	NN	O	B-DNA
DNA	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Susceptibility	NN	O	O
to	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
down	NN	O	O
regulation	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	O
I	NN	O	O
expression	NN	O	O
in	NN	O	O
adenovirus	NN	O	B-cell_line
12	NN	O	I-cell_line
transformed	NN	O	I-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
different	NN	O	O
E1A	NN	O	B-DNA
domains	NN	O	I-DNA
.	NN	O	O

All	NN	O	O
human	NN	O	O
adenoviruses	NN	O	O
transform	NN	O	O
rodent	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
cells	NN	O	O
transformed	NN	O	O
by	NN	O	O
serotypes	NN	O	O
belonging	NN	O	O
to	NN	O	O
subgroups	NN	O	O
A	NN	O	O
(	NN	O	O
Ad12	NN	O	O
)	NN	O	O
and	NN	O	O
B	NN	O	O
(	NN	O	O
Ad3	NN	O	O
)	NN	O	O
are	NN	O	O
tumorigenic	NN	O	O
for	NN	O	O
immunocompetent	NN	O	O
animals	NN	O	O
.	NN	O	O

In	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
MHC-class	NN	O	B-protein
I	NN	O	I-protein
antigens	NN	O	I-protein
is	NN	O	O
repressed	NN	O	O
and	NN	O	O
might	NN	O	O
allow	NN	O	O
them	NN	O	O
to	NN	O	O
escape	NN	O	O
from	NN	O	O
recognition	NN	O	O
by	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
CTL	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
to	NN	O	O
develop	NN	O	O
in	NN	O	O
tumor	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
these	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
appear	NN	O	O
resistant	NN	O	O
to	NN	O	O
lysis	NN	O	O
by	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
E1A	NN	O	B-DNA
domain	NN	O	I-DNA
(	NN	O	O
s	NN	O	O
)	NN	O	O
responsible	NN	O	O
for	NN	O	O
these	NN	O	O
properties	NN	O	O
several	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
created	NN	O	O
by	NN	O	O
transforming	NN	O	O
baby	NN	O	B-cell_line
rat	NN	O	I-cell_line
kidney	NN	O	I-cell_line
(	NN	O	I-cell_line
BRK	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
plasmids	NN	O	B-DNA
expressing	NN	O	O
different	NN	O	O
Ad2/Ad12	NN	O	B-protein
hybrid	NN	O	I-protein
E1A	NN	O	I-protein
gene	NN	O	I-protein
products	NN	O	I-protein
.	NN	O	O

The	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
1	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
inhibited	NN	O	O
in	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
Ad12	NN	O	B-protein
13S	NN	O	I-protein
mRNA	NN	O	I-protein
product	NN	O	I-protein
and	NN	O	O
in	NN	O	O
cells	NN	O	O
transformed	NN	O	O
with	NN	O	O
Ad2/Ad12	NN	O	B-protein
hybrid	NN	O	I-protein
E1A	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
harboring	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
part	NN	O	I-protein
of	NN	O	O
the	NN	O	O
conserved	NN	O	B-DNA
region	NN	O	I-DNA
(	NN	O	I-DNA
CR	NN	O	I-DNA
)	NN	O	I-DNA
3	NN	O	I-DNA
of	NN	O	O
Ad12	NN	O	O
.	NN	O	O

Susceptibility	NN	O	O
of	NN	O	O
these	NN	O	O
transformed	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
to	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
determined	NN	O	O
by	NN	O	O
cytolytic	NN	O	O
assays	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
obtained	NN	O	O
suggest	NN	O	O
that	NN	O	O
two	NN	O	O
Ad12	NN	O	B-DNA
E1A	NN	O	I-DNA
domains	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
to	NN	O	O
induce	NN	O	O
resistance	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
to	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Cell	NN	O	O
specific	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
Bruton	NN	O	I-DNA
's	NN	O	I-DNA
agammaglobulinemia	NN	O	I-DNA
tyrosine	NN	O	I-DNA
kinase	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
Btk	NN	O	B-protein
)	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
Sp1-	NN	O	B-protein
and	NN	O	O
Spi-1/PU.1-family	NN	O	B-protein
members	NN	O	I-protein
.	NN	O	O

Bruton	NN	O	B-protein
's	NN	O	I-protein
agammaglobulinemia	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
Btk	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
disease	NN	O	O
X-linked	NN	O	O
agammaglobulinemia	NN	O	O
(	NN	O	O
XLA	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
all	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
T-cells	NN	O	B-cell_type
and	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

For	NN	O	O
this	NN	O	O
expression	NN	O	O
pattern	NN	O	O
the	NN	O	O
first	NN	O	O
280	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
transcriptional	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
seems	NN	O	O
to	NN	O	O
be	NN	O	O
sufficient	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
footprinting	NN	O	O
analysis	NN	O	O
within	NN	O	O
this	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
revealed	NN	O	O
two	NN	O	O
Sp1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
PU-box	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Spi-1/PU.1	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
closely	NN	O	O
related	NN	O	O
factor	NN	O	O
Spi-B	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
PU-box	NN	O	B-DNA
in	NN	O	O
B-cells	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
erythroleukemia	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
K562	NN	O	I-cell_line
,	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
Spi-B	NN	O	B-protein
,	NN	O	O
only	NN	O	O
PU.1	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
Btk	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
either	NN	O	O
site	NN	O	O
reduced	NN	O	O
the	NN	O	O
expression	NN	O	O
in	NN	O	O
transient	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
PU	NN	O	B-DNA
box	NN	O	I-DNA
had	NN	O	O
no	NN	O	O
effect	NN	O	O
in	NN	O	O
the	NN	O	O
T-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
,	NN	O	O
where	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
Spi-1	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
Spi-B	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
PU.1	NN	O	B-protein
were	NN	O	O
able	NN	O	O
to	NN	O	O
transactivate	NN	O	O
Btk	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

In	NN	O	O
fetal	NN	O	O
liver	NN	O	O
of	NN	O	O
PU.1-/-	NN	O	O
mice	NN	O	O
,	NN	O	O
which	NN	O	O
lack	NN	O	O
lymphoid	NN	O	B-cell_type
and	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
expression	NN	O	O
of	NN	O	O
Btk	NN	O	B-protein
was	NN	O	O
reduced	NN	O	O
two-	NN	O	O
to	NN	O	O
threefold	NN	O	O
but	NN	O	O
not	NN	O	O
abolished	NN	O	O
.	NN	O	O

Collectively	NN	O	O
this	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Btk	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
combined	NN	O	O
action	NN	O	O
of	NN	O	O
Sp1-	NN	O	B-protein
and	NN	O	I-protein
PU.1-family	NN	O	I-protein
members	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

T	NN	O	O
cell	NN	O	O
response	NN	O	O
to	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
transactivators	NN	O	I-protein
in	NN	O	O
chronic	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
.	NN	O	O

Rheumatoid	NN	O	O
arthritis	NN	O	O
is	NN	O	O
a	NN	O	O
multistep	NN	O	O
disorder	NN	O	O
associated	NN	O	O
with	NN	O	O
autoimmune	NN	O	O
features	NN	O	O
of	NN	O	O
yet	NN	O	O
unknown	NN	O	O
etiology	NN	O	O
.	NN	O	O

Implication	NN	O	O
of	NN	O	O
viruses	NN	O	O
such	NN	O	O
as	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
in	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
pathogenesis	NN	O	O
has	NN	O	O
been	NN	O	O
suspected	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
several	NN	O	O
indirect	NN	O	O
observations	NN	O	O
,	NN	O	O
but	NN	O	O
thus	NN	O	O
far	NN	O	O
,	NN	O	O
a	NN	O	O
direct	NN	O	O
link	NN	O	O
between	NN	O	O
EBV	NN	O	O
and	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
provided	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
large	NN	O	O
fraction	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
infiltrating	NN	O	O
affected	NN	O	O
joints	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
chronic	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
recognizes	NN	O	O
two	NN	O	O
EBV	NN	O	B-protein
transactivators	NN	O	I-protein
(	NN	O	O
BZLF1	NN	O	B-protein
and	NN	O	O
BMLF1	NN	O	B-protein
)	NN	O	O
in	NN	O	O
a	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
-restricted	NN	O	O
fashion	NN	O	O
.	NN	O	O

Responses	NN	O	O
to	NN	O	O
these	NN	O	O
EBV	NN	O	B-protein
antigens	NN	O	I-protein
by	NN	O	O
synovial	NN	O	O
lymphocytes	NN	O	O
from	NN	O	O
several	NN	O	O
other	NN	O	O
chronic	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
patients	NN	O	O
were	NN	O	O
readily	NN	O	O
detectable	NN	O	O
.	NN	O	O

Thus	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
direct	NN	O	O
contribution	NN	O	O
of	NN	O	O
EBV	NN	O	O
to	NN	O	O
chronic	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

They	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
against	NN	O	O
EBV	NN	O	B-protein
transactivating	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
which	NN	O	O
might	NN	O	O
be	NN	O	O
central	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
virus	NN	O	O
reactivation	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD14	NN	O	B-protein
-mediated	NN	O	O
signal	NN	O	O
pathway	NN	O	O
of	NN	O	O
Porphyromonas	NN	O	O
gingivalis	NN	O	O
lipopolysaccharide	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
gingival	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

Lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
induces	NN	O	O
expression	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
in	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
via	NN	O	O
CD14	NN	O	B-protein
,	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
LPS	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
expressed	NN	O	O
predominantly	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
Porphyromonas	NN	O	O
gingivalis	NN	O	O
LPS	NN	O	O
(	NN	O	O
P-LPS	NN	O	O
)	NN	O	O
also	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
gingival	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
important	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
CD14	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
gingival	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
and	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
P-LPS-mediated	NN	O	O
signal-transducing	NN	O	B-protein
mechanism	NN	O	O
in	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
unexpectedly	NN	O	O
by	NN	O	O
immunohistochemical	NN	O	O
,	NN	O	O
Western	NN	O	O
blotting	NN	O	O
(	NN	O	O
immunoblotting	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
Northern	NN	O	O
(	NN	O	O
RNA	NN	O	O
)	NN	O	O
blotting	NN	O	O
assays	NN	O	O
that	NN	O	O
CD14	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
high	NN	O	O
density	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
gingival	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

P-LPS-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte	NN	O	B-DNA
chemoattractant	NN	O	I-DNA
protein	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	I-DNA
MCP-1	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
cells	NN	O	O
was	NN	O	O
inhibited	NN	O	O
markedly	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
anti-human	NN	O	O
CD14	NN	O	B-protein
antibody	NN	O	O
and	NN	O	O
was	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
by	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
.	NN	O	O

The	NN	O	O
inhibitor	NN	O	O
also	NN	O	O
dramatically	NN	O	O
inhibited	NN	O	O
monocyte	NN	O	O
chemotactic	NN	O	O
activity	NN	O	O
of	NN	O	O
and	NN	O	O
MCP-1	NN	O	B-protein
production	NN	O	O
by	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
P-LPS-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
MCP-1	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
cells	NN	O	O
also	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
two	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
curcumin	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
pyrolidine	NN	O	O
dithiocarbamate	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Both	NN	O	O
inhibitors	NN	O	O
inhibited	NN	O	O
monocyte	NN	O	O
chemotactic	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
culture	NN	O	O
supernatant	NN	O	O
of	NN	O	O
P-LPS-treated	NN	O	O
cells	NN	O	O
.	NN	O	O

Gel	NN	O	O
shift	NN	O	O
mobility	NN	O	O
assay	NN	O	O
showed	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
contents	NN	O	O
in	NN	O	O
P-LPS-treated	NN	O	O
cells	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CD14	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
gingival	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
and	NN	O	O
to	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
signal-transducing	NN	O	O
pathway	NN	O	O
of	NN	O	O
P-LPS	NN	O	O
in	NN	O	O
the	NN	O	O
cells	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
CD14	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

rel	NN	O	B-protein
Is	NN	O	O
rapidly	NN	O	O
tyrosine-phosphorylated	NN	O	O
following	NN	O	O
granulocyte-colony	NN	O	B-protein
stimulating	NN	O	I-protein
factor	NN	O	I-protein
treatment	NN	O	O
of	NN	O	O
human	NN	O	O
neutrophils	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
with	NN	O	O
granulocyte-colony	NN	O	B-protein
stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
G-CSF	NN	O	B-protein
)	NN	O	O
results	NN	O	O
in	NN	O	O
an	NN	O	O
enhanced	NN	O	O
respiratory	NN	O	O
burst	NN	O	O
,	NN	O	O
prolonged	NN	O	O
survival	NN	O	O
,	NN	O	O
and	NN	O	O
increased	NN	O	O
tumor	NN	O	O
cell	NN	O	O
killing	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
G-CSF	NN	O	B-protein
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
binding	NN	O	O
to	NN	O	O
specific	NN	O	O
,	NN	O	O
high	NN	O	O
affinity	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

G-CSF	NN	O	B-protein
receptors	NN	O	I-protein
lack	NN	O	O
intrinsic	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
but	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
rapid	NN	O	O
induction	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
activity	NN	O	O
.	NN	O	O

Antiphosphotyrosine	NN	O	O
immunoblots	NN	O	O
of	NN	O	O
whole	NN	O	O
cell	NN	O	O
lysates	NN	O	O
prepared	NN	O	O
from	NN	O	O
neutrophils	NN	O	B-cell_type
show	NN	O	O
that	NN	O	O
the	NN	O	O
G-CSF	NN	O	B-protein
rapidly	NN	O	O
induces	NN	O	O
prominent	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
a	NN	O	O
relative	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
80	NN	O	O
kDa	NN	O	O
.	NN	O	O

Using	NN	O	O
monospecific	NN	O	B-protein
antibodies	NN	O	I-protein
,	NN	O	O
the	NN	O	O
80-kDa	NN	O	O
tyrosine-phosphorylated	NN	O	B-protein
protein	NN	O	I-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
p80c-rel	NN	O	B-protein
,	NN	O	O
a	NN	O	O
proto-oncogene	NN	O	B-DNA
belonging	NN	O	O
to	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
regulators	NN	O	I-protein
which	NN	O	O
include	NN	O	O
NF-kB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
p80c-rel	NN	O	B-protein
was	NN	O	O
unique	NN	O	O
to	NN	O	O
G-CSF	NN	O	B-protein
in	NN	O	O
that	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony	NN	O	I-protein
stimulating	NN	O	I-protein
factor	NN	O	I-protein
which	NN	O	O
also	NN	O	O
stimulates	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
does	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
p80c-rel	NN	O	B-protein
.	NN	O	O

The	NN	O	O
consequences	NN	O	O
of	NN	O	O
p80c-rel	NN	O	B-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
are	NN	O	O
not	NN	O	O
yet	NN	O	O
known	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
tyrosine-phosphorylated	NN	O	O
p80c-rel	NN	O	B-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
,	NN	O	O
and	NN	O	O
G-CSF	NN	O	B-protein
stimulation	NN	O	O
results	NN	O	O
in	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
p80c-rel	NN	O	B-protein
which	NN	O	O
binds	NN	O	O
to	NN	O	O
DNA	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
biochemical	NN	O	O
events	NN	O	O
which	NN	O	O
occurs	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
following	NN	O	O
G-CSF	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
leads	NN	O	O
directly	NN	O	O
to	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
p80c-rel	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activated	NN	O	O
by	NN	O	O
G-CSF	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
directly	NN	O	O
transduce	NN	O	O
a	NN	O	O
signal	NN	O	O
to	NN	O	O
a	NN	O	O
protein	NN	O	O
which	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
regulator	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Patterns	NN	O	O
of	NN	O	O
Pan	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
role	NN	O	O
of	NN	O	O
Pan	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
endocrine	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
complex	NN	O	O
formation	NN	O	O
.	NN	O	O

The	NN	O	O
Pan	NN	O	B-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
distinct	NN	O	O
transcripts	NN	O	O
,	NN	O	O
Pan-1	NN	O	B-protein
and	NN	O	O
Pan-2	NN	O	B-protein
(	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
E47	NN	O	B-protein
and	NN	O	O
E12	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
,	NN	O	O
by	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
alternative	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analyses	NN	O	O
performed	NN	O	O
on	NN	O	O
rat	NN	O	O
and	NN	O	O
mouse	NN	O	O
tissues	NN	O	O
have	NN	O	O
detected	NN	O	O
ubiquitously	NN	O	O
expressed	NN	O	O
Pan	NN	O	O
transcripts	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
abundance	NN	O	O
,	NN	O	O
distribution	NN	O	O
,	NN	O	O
and	NN	O	O
form	NN	O	O
of	NN	O	O
Pan	NN	O	B-protein
proteins	NN	O	I-protein
have	NN	O	O
not	NN	O	O
been	NN	O	O
clearly	NN	O	O
defined	NN	O	O
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
representing	NN	O	O
endocrine	NN	O	B-cell_type
,	NN	O	I-cell_type
fibroblast	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
lineages	NN	O	I-cell_type
using	NN	O	O
polyclonal	NN	O	O
antisera	NN	O	O
to	NN	O	O
detect	NN	O	O
E2A	NN	O	B-protein
proteins	NN	O	I-protein
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
significant	NN	O	O
E2A	NN	O	B-protein
protein	NN	O	I-protein
expression	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
B-lymphocytes	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
,	NN	O	O
Yae	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
specific	NN	O	O
for	NN	O	O
Pan/E2A	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
and	NN	O	O
have	NN	O	O
used	NN	O	O
the	NN	O	O
Yae	NN	O	B-protein
antibody	NN	O	I-protein
to	NN	O	O
examine	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
endocrine	NN	O	B-cell_type
and	NN	O	I-cell_type
nonendocrine	NN	O	I-cell_type
cell	NN	O	I-cell_type
lineages	NN	O	I-cell_type
for	NN	O	O
differences	NN	O	O
in	NN	O	O
Pan/	NN	O	O
E2A	NN	O	B-protein
protein	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
subcellular	NN	O	O
localization	NN	O	O
,	NN	O	O
and	NN	O	O
heteromeric	NN	O	O
complex	NN	O	O
formation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
previous	NN	O	O
results	NN	O	O
obtained	NN	O	O
using	NN	O	O
polyclonal	NN	O	O
antiseras	NN	O	O
to	NN	O	O
detect	NN	O	O
Pan/	NN	O	O
E2A	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
we	NN	O	O
report	NN	O	O
comparable	NN	O	O
levels	NN	O	O
of	NN	O	O
Pan	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
GH/PRL-	NN	O	B-cell_line
and	NN	O	I-cell_line
insulin-producing	NN	O	I-cell_line
,	NN	O	I-cell_line
B-	NN	O	I-cell_line
and	NN	O	I-cell_line
T-lymphocyte	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

IEF-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
pancreatic	NN	O	O
beta-cell	NN	O	O
type-specific	NN	O	O
complex	NN	O	O
believed	NN	O	O
to	NN	O	O
regulate	NN	O	O
insulin	NN	O	O
expression	NN	O	O
,	NN	O	O
is	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
consist	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
distinct	NN	O	O
species	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
does	NN	O	O
not	NN	O	O
contain	NN	O	O
Pan	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
postulated	NN	O	O
that	NN	O	O
pituitary	NN	O	B-cell_type
endocrine	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
pancreatic	NN	O	B-cell_type
endocrine	NN	O	I-cell_type
beta-cells	NN	O	I-cell_type
share	NN	O	O
identical	NN	O	O
Pan/E2A	NN	O	B-protein
complexes	NN	O	I-protein
,	NN	O	O
native-Western	NN	O	O
analyses	NN	O	O
of	NN	O	O
pituitary	NN	O	O
and	NN	O	O
endocrine	NN	O	B-cell_type
beta-cells	NN	O	I-cell_type
detect	NN	O	O
Pan	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
distinct	NN	O	O
cell	NN	O	B-protein
type-specific	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Visualization	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
p50	NN	O	I-protein
subunit	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
follicular	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
germinal	NN	O	O
centers	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
50	NN	O	B-protein
kDa/65	NN	O	I-protein
kDa	NN	O	I-protein
(	NN	O	I-protein
p50/p65	NN	O	I-protein
)	NN	O	I-protein
heterodimer	NN	O	I-protein
,	NN	O	O
is	NN	O	O
a	NN	O	O
ubiquitous	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
positive	NN	O	O
regulation	NN	O	O
of	NN	O	O
various	NN	O	O
immune	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
related	NN	O	O
to	NN	O	O
a	NN	O	O
particular	NN	O	O
cell	NN	O	O
type	NN	O	O
and/or	NN	O	O
differentiation	NN	O	O
step	NN	O	O
during	NN	O	O
immunopoiesis	NN	O	O
.	NN	O	O

Using	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
on	NN	O	O
sections	NN	O	O
from	NN	O	O
non	NN	O	O
HIV	NN	O	O
hyperplastic	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
gene	NN	O	O
of	NN	O	O
the	NN	O	O
105	NN	O	B-protein
kDa	NN	O	I-protein
precursor	NN	O	I-protein
of	NN	O	O
p50	NN	O	B-protein
was	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
the	NN	O	O
light	NN	O	O
zone	NN	O	O
of	NN	O	O
germinal	NN	O	O
centers	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
network	NN	O	O
aspect	NN	O	O
,	NN	O	O
which	NN	O	O
suggested	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
follicular	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
FDC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

By	NN	O	O
immunohistochemistry	NN	O	O
,	NN	O	O
p50	NN	O	B-protein
protein	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
and	NN	O	O
nucleus	NN	O	O
of	NN	O	O
FDC	NN	O	B-cell_type
,	NN	O	O
confirming	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
FDC	NN	O	B-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
p50	NN	O	B-protein
protein	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
all	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
we	NN	O	O
focused	NN	O	O
our	NN	O	O
study	NN	O	O
on	NN	O	O
isolated	NN	O	O
FDC	NN	O	B-cell_type
clusters	NN	O	O
from	NN	O	O
normal	NN	O	O
tonsils	NN	O	O
.	NN	O	O

As	NN	O	O
showed	NN	O	O
on	NN	O	O
tissue	NN	O	O
sections	NN	O	O
,	NN	O	O
we	NN	O	O
detected	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
in	NN	O	O
both	NN	O	O
cytoplasm	NN	O	O
and	NN	O	O
nucleus	NN	O	O
of	NN	O	O
FDC	NN	O	B-cell_type
.	NN	O	O

Nuclei	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
FDC	NN	O	B-cell_type
clusters	NN	O	O
were	NN	O	O
negative	NN	O	O
.	NN	O	O

We	NN	O	O
next	NN	O	O
studied	NN	O	O
p65	NN	O	O
and	NN	O	O
c-Rel	NN	O	O
protein	NN	O	O
expression	NN	O	O
in	NN	O	O
FDC	NN	O	B-cell_type
clusters	NN	O	O
.	NN	O	O

p65	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
FDC	NN	O	B-cell_type
,	NN	O	O
whereas	NN	O	O
nuclei	NN	O	O
were	NN	O	O
negative	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
p65	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
some	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

c-Rel	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
and	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
FDC	NN	O	B-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicated	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
T	NN	O	O
cell-dependent	NN	O	O
B	NN	O	O
cell	NN	O	O
immunopoiesis	NN	O	O
occurring	NN	O	O
in	NN	O	O
FDC	NN	O	B-cell_type
clusters	NN	O	O
,	NN	O	O
p50	NN	O	B-protein
is	NN	O	O
mainly	NN	O	O
related	NN	O	O
to	NN	O	O
FDC	NN	O	B-cell_type
with	NN	O	O
a	NN	O	O
massive	NN	O	O
overexpression	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
,	NN	O	O
whereas	NN	O	O
p65	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
scattered	NN	O	O
manner	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
c-Rel	NN	O	O
protein	NN	O	O
exclusively	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
FDC	NN	O	B-cell_type
clusters	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
subunits	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
the	NN	O	O
c-Rel	NN	O	B-protein
protein	NN	O	I-protein
have	NN	O	O
different	NN	O	O
roles	NN	O	O
in	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
during	NN	O	O
B	NN	O	O
cell	NN	O	O
immunopoiesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
NF-ATc	NN	O	B-protein
and	NN	O	O
NF-ATp	NN	O	B-protein
:	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
components	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

Present	NN	O	O
evidence	NN	O	O
indicates	NN	O	O
a	NN	O	O
pathway	NN	O	O
of	NN	O	O
signal	NN	O	O
transmission	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
is	NN	O	O
outlined	NN	O	O
in	NN	O	O
figure	NN	O	O
1	NN	O	O
.	NN	O	O

The	NN	O	O
elevation	NN	O	O
in	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
that	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
interactions	NN	O	O
at	NN	O	O
the	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
leads	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
calcium-dependent	NN	O	B-protein
phosphatase	NN	O	I-protein
calcineurin	NN	O	B-protein
.	NN	O	O

This	NN	O	O
in	NN	O	O
turn	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
nuclear	NN	O	O
association	NN	O	O
of	NN	O	O
the	NN	O	O
cytosolic	NN	O	B-protein
component	NN	O	I-protein
of	NN	O	O
NF-ATc	NN	O	B-protein
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
and	NN	O	O
the	NN	O	O
nuclear	NN	O	O
import	NN	O	O
of	NN	O	O
NF-ATc	NN	O	B-protein
can	NN	O	O
both	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
or	NN	O	O
FK506	NN	O	O
in	NN	O	O
complex	NN	O	O
with	NN	O	O
their	NN	O	O
respective	NN	O	O
immunophilins	NN	O	B-protein
.	NN	O	O

Once	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
NF-ATc	NN	O	B-protein
interacts	NN	O	O
with	NN	O	O
NF-ATn	NN	O	B-protein
to	NN	O	O
form	NN	O	O
an	NN	O	O
active	NN	O	B-protein
transcriptional	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

NF-ATn	NN	O	B-protein
is	NN	O	O
a	NN	O	O
ubiquitous	NN	O	O
protein	NN	O	O
,	NN	O	O
can	NN	O	O
be	NN	O	O
synthesized	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
PMA	NN	O	O
,	NN	O	O
and	NN	O	O
has	NN	O	O
many	NN	O	O
similarities	NN	O	O
to	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
NF-ATc	NN	O	B-protein
enters	NN	O	O
the	NN	O	O
nucleus	NN	O	O
is	NN	O	O
unknown	NN	O	O
,	NN	O	O
and	NN	O	O
although	NN	O	O
it	NN	O	O
appears	NN	O	O
to	NN	O	O
require	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
NF-ATc	NN	O	B-protein
has	NN	O	O
not	NN	O	O
yet	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
in	NN	O	O
vivo	NN	O	O
substrate	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

Alternative	NN	O	O
mechanisms	NN	O	O
include	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
NF-ATc	NN	O	B-protein
operates	NN	O	O
on	NN	O	O
some	NN	O	O
cytoplasmic	NN	O	B-protein
anchor	NN	O	I-protein
or	NN	O	O
that	NN	O	O
other	NN	O	O
proteins	NN	O	O
that	NN	O	O
are	NN	O	O
controlled	NN	O	O
by	NN	O	O
calcineurin	NN	O	B-protein
carry	NN	O	O
out	NN	O	O
the	NN	O	O
nuclear	NN	O	O
import	NN	O	O
of	NN	O	O
NF-ATc	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
NF-ATp	NN	O	B-protein
copurifies	NN	O	O
with	NN	O	O
NF-ATc	NN	O	B-protein
,	NN	O	O
there	NN	O	O
is	NN	O	O
as	NN	O	O
yet	NN	O	O
no	NN	O	O
understanding	NN	O	O
of	NN	O	O
how	NN	O	O
NF-ATp	NN	O	B-protein
is	NN	O	O
functioning	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Now	NN	O	O
that	NN	O	O
these	NN	O	O
proteins	NN	O	O
are	NN	O	O
purified	NN	O	O
and	NN	O	O
cloned	NN	O	O
,	NN	O	O
the	NN	O	O
major	NN	O	O
goals	NN	O	O
will	NN	O	O
be	NN	O	O
to	NN	O	O
understand	NN	O	O
their	NN	O	O
role	NN	O	O
and	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
other	NN	O	O
family	NN	O	O
members	NN	O	O
in	NN	O	O
thymic	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
by	NN	O	O
low-density	NN	O	B-protein
lipoprotein	NN	O	I-protein
.	NN	O	O

Low-density	NN	O	B-protein
lipoprotein	NN	O	I-protein
(	NN	O	O
LDL	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
well-established	NN	O	O
risk	NN	O	O
factor	NN	O	O
for	NN	O	O
atherosclerosis	NN	O	O
.	NN	O	O

When	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
are	NN	O	O
incubated	NN	O	O
with	NN	O	O
this	NN	O	O
lipoprotein	NN	O	O
in	NN	O	O
pathophysiologic	NN	O	O
amounts	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
are	NN	O	O
activated	NN	O	O
.	NN	O	O

Among	NN	O	O
the	NN	O	O
documented	NN	O	O
cellular	NN	O	O
responses	NN	O	O
to	NN	O	O
LDL	NN	O	B-protein
is	NN	O	O
increased	NN	O	O
recruitment	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
which	NN	O	O
are	NN	O	O
believed	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
promoting	NN	O	O
intimal	NN	O	O
plaque	NN	O	O
formation	NN	O	O
.	NN	O	O

The	NN	O	O
findings	NN	O	O
presented	NN	O	O
here	NN	O	O
link	NN	O	O
an	NN	O	O
atheogenic	NN	O	B-protein
lipoprotein	NN	O	I-protein
,	NN	O	O
LDL	NN	O	B-protein
,	NN	O	O
with	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
an	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
important	NN	O	O
in	NN	O	O
atherogenesis	NN	O	O
Human	NN	O	O
LDL	NN	O	B-protein
induces	NN	O	O
the	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
transcriptionally	NN	O	O
with	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
VCAM	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
effect	NN	O	O
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
anti-VCAM	NN	O	B-protein
antibodies	NN	O	I-protein
.	NN	O	O

After	NN	O	O
a	NN	O	O
2-day	NN	O	O
incubation	NN	O	O
in	NN	O	O
LDL	NN	O	B-protein
,	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
key	NN	O	O
oxidative-stress	NN	O	O
sensor	NN	O	O
for	NN	O	O
VCAM	NN	O	B-protein
regulation	NN	O	O
,	NN	O	O
remains	NN	O	O
at	NN	O	O
basal	NN	O	O
level	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
activities	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
GATA	NN	O	B-protein
,	NN	O	O
on	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
are	NN	O	O
increased	NN	O	O
by	NN	O	O
LDL	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
LDL	NN	O	B-protein
-enhanced	NN	O	O
endothelial	NN	O	O
recruitment	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
is	NN	O	O
attributed	NN	O	O
to	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
which	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
through	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
GATA	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
identify	NN	O	O
LDL	NN	O	B-protein
as	NN	O	O
a	NN	O	O
VCAM	NN	O	B-protein
-inducer	NN	O	O
possibly	NN	O	O
distinct	NN	O	O
from	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
endotoxin	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
transcription	NN	O	O
factor	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
DNA	NN	O	I-DNA
sequence	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
various	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Although	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IgE	NN	O	O
synthesis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
we	NN	O	O
first	NN	O	O
examined	NN	O	O
the	NN	O	O
participation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
germline	NN	O	B-cell_line
C	NN	O	I-cell_line
epsilon	NN	O	I-cell_line
transcription	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
Burkitt	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
DND39	NN	O	B-cell_line
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
DND39	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
IL-4	NN	O	B-protein
or	NN	O	O
anti-CD40	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
mAb	NN	O	B-protein
)	NN	O	O
activated	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
and	NN	O	O
subsequently	NN	O	O
induced	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
was	NN	O	O
identified	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

n-Acetyl-L-cysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
antioxidant	NN	O	O
,	NN	O	O
blocked	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
caused	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
by	NN	O	O
anti-CD40	NN	O	B-protein
mAb	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-4-driven	NN	O	B-cell_line
germline	NN	O	I-cell_line
C	NN	O	I-cell_line
epsilon	NN	O	I-cell_line
transcription	NN	O	O
by	NN	O	O
NAC	NN	O	O
was	NN	O	O
not	NN	O	O
sufficient	NN	O	O
,	NN	O	O
the	NN	O	O
agent	NN	O	O
remarkably	NN	O	O
diminished	NN	O	O
anti-CD40	NN	O	B-protein
mAb	NN	O	I-protein
-mediated	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
germline	NN	O	B-DNA
C	NN	O	O
epsilon	NN	O	O
transcription	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
NAC	NN	O	O
on	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
human	NN	O	O
normal	NN	O	O
B	NN	O	O
cells	NN	O	O
costimulated	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
anti-CD40	NN	O	B-protein
mAb	NN	O	I-protein
.	NN	O	O

NAC	NN	O	O
was	NN	O	O
effective	NN	O	O
in	NN	O	O
inhibiting	NN	O	O
mature	NN	O	O
C	NN	O	O
epsilon	NN	O	O
transcription	NN	O	O
and	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
the	NN	O	O
T	NN	O	O
cell-independent	NN	O	O
culture	NN	O	O
system	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
NAC	NN	O	O
did	NN	O	O
not	NN	O	O
significantly	NN	O	O
affect	NN	O	O
the	NN	O	O
spontaneous	NN	O	O
production	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
by	NN	O	O
atopic	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
is	NN	O	O
commonly	NN	O	O
inducible	NN	O	O
in	NN	O	O
DND39	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
anti-CD40	NN	O	B-protein
mAb	NN	O	I-protein
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
sensitive	NN	O	O
to	NN	O	O
NAC	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Prostaglandin	NN	O	O
E2	NN	O	O
induction	NN	O	O
of	NN	O	O
binding	NN	O	O
activity	NN	O	O
to	NN	O	O
CRE	NN	O	B-DNA
and	NN	O	O
AP-2	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Prostaglandins	NN	O	O
of	NN	O	O
the	NN	O	O
E	NN	O	O
series	NN	O	O
are	NN	O	O
immunomodulatory	NN	O	O
agents	NN	O	O
which	NN	O	O
exert	NN	O	O
inhibitory	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
stimulatory	NN	O	O
effects	NN	O	O
on	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

Since	NN	O	O
it	NN	O	O
is	NN	O	O
known	NN	O	O
that	NN	O	O
PGE2	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
increase	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
it	NN	O	O
can	NN	O	O
affect	NN	O	O
gene	NN	O	O
expression	NN	O	O
through	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
which	NN	O	O
bind	NN	O	O
enhancer	NN	O	O
elements	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	O
regions	NN	O	O
of	NN	O	O
cAMP-regulated	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
a	NN	O	O
short	NN	O	O
treatment	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
PGE2	NN	O	O
induces	NN	O	O
specific	NN	O	O
binding	NN	O	O
activity	NN	O	O
to	NN	O	O
CRE	NN	O	B-DNA
and	NN	O	O
AP-2	NN	O	B-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
AP-1	NN	O	B-DNA
,	NN	O	I-DNA
DNA	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Since	NN	O	O
the	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
protein	NN	O	O
phosphatase	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
prolongs	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
phosphorylation	NN	O	O
events	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
occur	NN	O	O
.	NN	O	O

This	NN	O	O
activity	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
increased	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
because	NN	O	O
forskolin	NN	O	O
and	NN	O	O
IBMX	NN	O	O
mimic	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
PGE2	NN	O	O
.	NN	O	O

More	NN	O	O
interestingly	NN	O	O
,	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
with	NN	O	O
CRE-CAT	NN	O	B-DNA
plasmide	NN	O	I-DNA
show	NN	O	O
that	NN	O	O
PGE2	NN	O	O
activates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
CRE-containing	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
positive	NN	O	O
role	NN	O	O
for	NN	O	O
PGE2	NN	O	O
on	NN	O	O
some	NN	O	O
immune	NN	O	O
functions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nasal	NN	O	B-cell_type
NK-	NN	O	I-cell_type
and	NN	O	I-cell_type
T-cell	NN	O	I-cell_type
lymphomas	NN	O	I-cell_type
share	NN	O	O
the	NN	O	O
same	NN	O	O
type	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
latency	NN	O	O
as	NN	O	O
nasopharyngeal	NN	O	O
carcinoma	NN	O	O
and	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

Nasal	NN	O	B-cell_type
T/NK-cell	NN	O	I-cell_type
lymphomas	NN	O	I-cell_type
can	NN	O	O
be	NN	O	O
further	NN	O	O
separated	NN	O	O
into	NN	O	O
those	NN	O	O
of	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
cell	NN	O	I-cell_type
lineage	NN	O	I-cell_type
or	NN	O	O
of	NN	O	O
T-cell	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
with	NN	O	O
differences	NN	O	O
in	NN	O	O
cellular	NN	O	O
phenotype	NN	O	O
,	NN	O	O
T-cell	NN	O	B-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
TcR	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
rearrangement	NN	O	O
and	NN	O	O
TcR	NN	O	O
transcript	NN	O	O
expression	NN	O	O
.	NN	O	O

Both	NN	O	O
NK-	NN	O	B-cell_type
and	NN	O	I-cell_type
T-cell	NN	O	I-cell_type
subtypes	NN	O	I-cell_type
are	NN	O	O
closely	NN	O	O
associated	NN	O	O
with	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
EBV	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
determined	NN	O	O
in	NN	O	O
23	NN	O	O
cases	NN	O	O
of	NN	O	O
nasal	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
NL	NN	O	O
)	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridisation	NN	O	O
(	NN	O	O
ISH	NN	O	O
)	NN	O	O
,	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
and	NN	O	O
immunohistochemistry	NN	O	O
(	NN	O	O
IH	NN	O	O
)	NN	O	O
.	NN	O	O

Of	NN	O	O
the	NN	O	O
23	NN	O	O
cases	NN	O	O
,	NN	O	O
19	NN	O	O
were	NN	O	O
classified	NN	O	O
as	NN	O	O
NK-cell	NN	O	O
and	NN	O	O
4	NN	O	O
as	NN	O	O
T-cell	NN	O	O
tumours	NN	O	O
.	NN	O	O

ISH	NN	O	O
for	NN	O	O
EBV-encoded	NN	O	B-RNA
small	NN	O	I-RNA
non-polyadenylated	NN	O	I-RNA
RNAs	NN	O	I-RNA
showed	NN	O	O
that	NN	O	O
all	NN	O	O
cases	NN	O	O
,	NN	O	O
whether	NN	O	O
NK	NN	O	O
or	NN	O	O
T	NN	O	O
,	NN	O	O
harboured	NN	O	O
EBV	NN	O	O
in	NN	O	O
virtually	NN	O	O
all	NN	O	O
tumour	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

RT-PCR	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
NL	NN	O	B-cell_type
of	NN	O	O
both	NN	O	O
subtypes	NN	O	O
expressed	NN	O	O
EBNAI	NN	O	B-protein
of	NN	O	O
the	NN	O	O
QUK	NN	O	O
splice	NN	O	O
pattern	NN	O	O
,	NN	O	O
the	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
LMP1	NN	O	B-protein
and	NN	O	I-protein
2	NN	O	I-protein
and	NN	O	O
the	NN	O	O
BamHI	NN	O	B-RNA
A	NN	O	I-RNA
rightward	NN	O	I-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
EBNA2	NN	O	B-RNA
mRNAs	NN	O	I-RNA
,	NN	O	O
compatible	NN	O	O
with	NN	O	O
the	NN	O	O
latency	NN	O	O
type	NN	O	O
II	NN	O	O
pattern	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
analysis	NN	O	O
of	NN	O	O
EBV	NN	O	O
protein	NN	O	O
expression	NN	O	O
by	NN	O	O
IH	NN	O	O
revealed	NN	O	O
a	NN	O	O
heterogeneous	NN	O	O
pattern	NN	O	O
of	NN	O	O
EBV	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
at	NN	O	O
the	NN	O	O
single-cell	NN	O	O
level	NN	O	O
consisting	NN	O	O
of	NN	O	O
both	NN	O	O
LMP1+	NN	O	B-cell_type
and	NN	O	I-cell_type
LMP1-	NN	O	I-cell_type
tumour	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
mixture	NN	O	O
of	NN	O	O
latency	NN	O	O
I	NN	O	O
and	NN	O	O
II	NN	O	O
.	NN	O	O

Although	NN	O	O
2	NN	O	O
early	NN	O	B-protein
lytic	NN	O	I-protein
transcripts	NN	O	I-protein
,	NN	O	O
BZLF1	NN	O	B-protein
and	NN	O	O
BHRF1	NN	O	B-protein
,	NN	O	O
were	NN	O	O
also	NN	O	O
detected	NN	O	O
in	NN	O	O
13	NN	O	O
and	NN	O	O
10	NN	O	O
cases	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-protein
staining	NN	O	O
in	NN	O	O
any	NN	O	O
case	NN	O	O
indicates	NN	O	O
that	NN	O	O
these	NN	O	O
lytic	NN	O	O
transcripts	NN	O	O
are	NN	O	O
most	NN	O	O
likely	NN	O	O
expressed	NN	O	O
by	NN	O	O
rare	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
biopsies	NN	O	O
entering	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
.	NN	O	O

The	NN	O	O
viral	NN	O	O
transcriptional	NN	O	O
pattern	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
nasopharyngeal	NN	O	O
carcinoma	NN	O	O
and	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
suggests	NN	O	O
that	NN	O	O
EBV	NN	O	O
can	NN	O	O
exploit	NN	O	O
common	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
diverse	NN	O	O
host	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Down-regulation	NN	O	O
of	NN	O	O
immunogenic	NN	O	B-protein
proteins	NN	O	I-protein
(	NN	O	O
EBNA2-EBNA6	NN	O	B-protein
)	NN	O	O
in	NN	O	O
nasal	NN	O	O
lymphoma	NN	O	O
may	NN	O	O
enable	NN	O	O
tumour	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
evade	NN	O	O
host	NN	O	O
cytotoxic	NN	O	O
T-cell	NN	O	O
surveillance	NN	O	O
.	NN	O	O

-DOCSTART-	O

Potent	NN	O	O
gene	NN	O	O
regulatory	NN	O	O
and	NN	O	O
antiproliferative	NN	O	O
activities	NN	O	O
of	NN	O	O
20-methyl	NN	O	O
analogues	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
dihydroxyvitamin	NN	O	O
D3	NN	O	O
.	NN	O	O

The	NN	O	O
biological	NN	O	O
active	NN	O	O
form	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
VD	NN	O	O
)	NN	O	O
,	NN	O	O
regulates	NN	O	O
cellular	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

This	NN	O	O
provides	NN	O	O
the	NN	O	O
hormone	NN	O	O
with	NN	O	O
an	NN	O	O
interesting	NN	O	O
therapeutic	NN	O	O
potential	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
hypercalcemia	NN	O	O
is	NN	O	O
a	NN	O	O
side	NN	O	O
effect	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
VD	NN	O	O
's	NN	O	O
classical	NN	O	O
action	NN	O	O
,	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
calcium	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

This	NN	O	O
made	NN	O	O
the	NN	O	O
need	NN	O	O
for	NN	O	O
VD	NN	O	O
analogues	NN	O	O
with	NN	O	O
selectively	NN	O	O
increased	NN	O	O
cell	NN	O	O
regulatory	NN	O	O
properties	NN	O	O
.	NN	O	O

Studies	NN	O	O
with	NN	O	O
20-epi	NN	O	O
analogues	NN	O	O
pointed	NN	O	O
out	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
the	NN	O	O
carbon-20	NN	O	O
position	NN	O	O
and	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
20-methyl	NN	O	O
derivatives	NN	O	O
of	NN	O	O
VD	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
the	NN	O	O
biological	NN	O	O
properties	NN	O	O
of	NN	O	O
the	NN	O	O
compounds	NN	O	O
ZK161422	NN	O	O
and	NN	O	O
ZK157202	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
20-methyl-	NN	O	O
and	NN	O	O
20-methyl-23-eneanalogues	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
analyzed	NN	O	O
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
VD	NN	O	O
.	NN	O	O

Both	NN	O	O
compounds	NN	O	O
show	NN	O	O
about	NN	O	O
2-fold	NN	O	O
lower	NN	O	O
affinity	NN	O	O
to	NN	O	O
the	NN	O	O
VD	NN	O	O
receptor	NN	O	O
(	NN	O	O
VDR	NN	O	O
)	NN	O	O
than	NN	O	O
VD	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
compared	NN	O	O
to	NN	O	O
VD	NN	O	O
,	NN	O	O
their	NN	O	O
antiproliferative	NN	O	O
effect	NN	O	O
is	NN	O	O
up	NN	O	O
to	NN	O	O
30-fold	NN	O	O
higher	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
even	NN	O	O
up	NN	O	O
to	NN	O	O
300-fold	NN	O	O
higher	NN	O	O
on	NN	O	O
human	NN	O	B-cell_line
breast	NN	O	I-cell_line
cancer	NN	O	I-cell_line
MCF-7	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Whereas	NN	O	O
the	NN	O	O
hypercalcemic	NN	O	O
effect	NN	O	O
for	NN	O	O
ZK157202	NN	O	O
is	NN	O	O
also	NN	O	O
increased	NN	O	O
10-fold	NN	O	O
,	NN	O	O
ZK161422	NN	O	O
has	NN	O	O
the	NN	O	O
same	NN	O	O
calcium-mobilizing	NN	O	O
potency	NN	O	O
as	NN	O	O
VD	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
ZK161422	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
ZK157202	NN	O	O
,	NN	O	O
showed	NN	O	O
preference	NN	O	O
for	NN	O	O
gene	NN	O	O
activation	NN	O	O
from	NN	O	O
a	NN	O	O
promoter	NN	O	B-DNA
carrying	NN	O	O
a	NN	O	O
VD	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
with	NN	O	O
a	NN	O	O
palindromic	NN	O	O
arrangement	NN	O	O
of	NN	O	O
two	NN	O	O
hexameric	NN	O	B-DNA
receptor	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
spaced	NN	O	O
by	NN	O	O
9	NN	O	O
nucleotides	NN	O	O
(	NN	O	O
IP9	NN	O	O
)	NN	O	O
rather	NN	O	O
than	NN	O	O
for	NN	O	O
activation	NN	O	O
from	NN	O	O
a	NN	O	O
response	NN	O	B-DNA
element	NN	O	I-DNA
formed	NN	O	O
by	NN	O	O
a	NN	O	O
direct	NN	O	O
repeat	NN	O	O
spaced	NN	O	O
by	NN	O	O
3	NN	O	O
nucleotides	NN	O	O
(	NN	O	O
DR3	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
supports	NN	O	O
a	NN	O	O
model	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
promoter	NN	O	O
selectivity	NN	O	O
reflects	NN	O	O
the	NN	O	O
selectively	NN	O	O
increased	NN	O	O
antiproliferative	NN	O	O
effect	NN	O	O
of	NN	O	O
VD	NN	O	O
analogues	NN	O	O
.	NN	O	O

-DOCSTART-	O

Calcium-dependent	NN	O	O
immediate-early	NN	O	B-DNA
gene	NN	O	I-DNA
induction	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
is	NN	O	O
negatively	NN	O	O
regulated	NN	O	O
by	NN	O	O
p21Ha-ras	NN	O	B-protein
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
immediate-early	NN	O	B-DNA
(	NN	O	I-DNA
IE	NN	O	I-DNA
)	NN	O	I-DNA
response	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
egr-1	NN	O	B-DNA
,	NN	O	O
c-fos	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	O
occurs	NN	O	O
rapidly	NN	O	O
after	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
process	NN	O	O
of	NN	O	O
activation	NN	O	O
involves	NN	O	O
calcium	NN	O	O
mobilization	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

p21	NN	O	B-protein
(	NN	O	I-protein
ras	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
a	NN	O	O
guanine	NN	O	B-protein
nucleotide	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
mediates	NN	O	O
T-cell	NN	O	O
signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
PKC	NN	O	B-protein
-dependent	NN	O	O
and	NN	O	O
PKC	NN	O	B-protein
-independent	NN	O	O
pathways	NN	O	O
.	NN	O	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
ras	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
calcium-dependent	NN	O	O
signals	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
through	NN	O	O
analysis	NN	O	O
of	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
inductions	NN	O	O
of	NN	O	O
the	NN	O	O
IE	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
calcium	NN	O	O
signals	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
activated	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
ras	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	O
and	NN	O	O
their	NN	O	O
correlated	NN	O	O
consequences	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
activated	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
ras	NN	O	I-protein
)	NN	O	I-protein
negatively	NN	O	O
regulated	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IE	NN	O	B-DNA
genes	NN	O	I-DNA
by	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
of	NN	O	O
calcium-activated	NN	O	O
IE	NN	O	B-DNA
gene	NN	O	I-DNA
induction	NN	O	O
was	NN	O	O
reversed	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
in	NN	O	O
this	NN	O	O
regulation	NN	O	O
.	NN	O	O

A	NN	O	O
later	NN	O	O
result	NN	O	O
of	NN	O	O
inhibition	NN	O	O
of	NN	O	O
this	NN	O	O
activation	NN	O	O
pathway	NN	O	O
by	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
ras	NN	O	I-protein
)	NN	O	I-protein
was	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
and	NN	O	O
subsequent	NN	O	O
coordinate	NN	O	O
reductions	NN	O	O
in	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
and	NN	O	O
protein	NN	O	O
production	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
p2l	NN	O	B-protein
(	NN	O	I-protein
ras	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
an	NN	O	O
essential	NN	O	O
mediator	NN	O	O
in	NN	O	O
generating	NN	O	O
not	NN	O	O
only	NN	O	O
positive	NN	O	O
but	NN	O	O
also	NN	O	O
negative	NN	O	O
modulatory	NN	O	O
mechanisms	NN	O	O
controlling	NN	O	O
the	NN	O	O
competence	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
inductive	NN	O	O
stimulations	NN	O	O
.	NN	O	O

-DOCSTART-	O

Calcineurin	NN	O	B-protein
acts	NN	O	O
in	NN	O	O
synergy	NN	O	O
with	NN	O	O
PMA	NN	O	O
to	NN	O	O
inactivate	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B/MAD3	NN	O	I-protein
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
consists	NN	O	O
of	NN	O	O
several	NN	O	O
independent	NN	O	O
T	NN	O	B-DNA
cell	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
TcR	NN	O	I-DNA
)	NN	O	I-DNA
responsive	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
promoters	NN	O	B-DNA
dependent	NN	O	O
on	NN	O	O
these	NN	O	O
elements	NN	O	O
is	NN	O	O
inhibitable	NN	O	O
by	NN	O	O
the	NN	O	O
immunosuppressants	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
and	NN	O	O
tacrolimus	NN	O	O
(	NN	O	O
FK-506	NN	O	O
)	NN	O	O
.	NN	O	O

Calcineurin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
Ca2+/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
phosphatase	NN	O	I-protein
,	NN	O	O
is	NN	O	O
the	NN	O	O
FK-506	NN	O	O
-and	NN	O	O
CsA-sensitive	NN	O	O
enzyme	NN	O	O
required	NN	O	O
for	NN	O	O
TcR	NN	O	O
mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
a	NN	O	O
constitutively	NN	O	O
active	NN	O	O
form	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
partially	NN	O	O
substitutes	NN	O	O
for	NN	O	O
the	NN	O	O
Ca2+	NN	O	O
co-stimulus	NN	O	O
required	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
elements	NN	O	I-DNA
IL-2A	NN	O	B-DNA
(	NN	O	O
which	NN	O	O
binds	NN	O	O
the	NN	O	O
factors	NN	O	O
OAP	NN	O	B-protein
and	NN	O	O
Oct-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IL-2E	NN	O	B-DNA
(	NN	O	O
which	NN	O	O
binds	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
completely	NN	O	O
substitutes	NN	O	O
for	NN	O	O
the	NN	O	O
Ca2+	NN	O	O
co-stimulus	NN	O	O
required	NN	O	O
to	NN	O	O
stimulate	NN	O	O
an	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
element	NN	O	O
.	NN	O	O

Calcineurin	NN	O	B-protein
stimulates	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
element	NN	O	I-DNA
by	NN	O	O
enhancing	NN	O	O
inactivation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B/MAD3	NN	O	I-protein
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
thereby	NN	O	O
increasing	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
nuclear	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
the	NN	O	O
first	NN	O	O
demonstration	NN	O	O
in	NN	O	O
vivo	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
phosphatase	NN	O	I-protein
can	NN	O	O
inactivate	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
one	NN	O	O
possible	NN	O	O
explanation	NN	O	O
for	NN	O	O
mechanism-based	NN	O	O
toxicities	NN	O	O
associated	NN	O	O
with	NN	O	O
FK-506	NN	O	O
and	NN	O	O
CsA	NN	O	O
by	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
these	NN	O	O
drugs	NN	O	O
can	NN	O	O
inhibit	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
virtually	NN	O	O
ubiquitous	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Detection	NN	O	O
of	NN	O	O
minimal	NN	O	O
residual	NN	O	O
disease	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
by	NN	O	O
RT-PCR	NN	O	O
:	NN	O	O
necessity	NN	O	O
of	NN	O	O
chemotherapy	NN	O	O
following	NN	O	O
ATRA	NN	O	O
therapy	NN	O	O
.	NN	O	O

The	NN	O	O
PML/RAR	NN	O	B-DNA
alpha	NN	O	I-DNA
fusion	NN	O	I-DNA
gene	NN	O	I-DNA
resulting	NN	O	O
from	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
15	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
translocation	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
specific	NN	O	O
marker	NN	O	O
for	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
bone	NN	O	O
marrow	NN	O	O
cells	NN	O	O
by	NN	O	O
reverse	NN	O	O
transcriptase-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
to	NN	O	O
detect	NN	O	O
residual	NN	O	O
PML/RAR	NN	O	B-cell_type
alpha	NN	O	I-cell_type
mRNA-containing	NN	O	I-cell_type
cells	NN	O	I-cell_type
following	NN	O	O
treatment	NN	O	O
with	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
and	NN	O	O
cytotoxic	NN	O	O
chemotherapy	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
APL	NN	O	O
.	NN	O	O

This	NN	O	O
RT-PCR	NN	O	O
assay	NN	O	O
can	NN	O	O
detect	NN	O	O
one	NN	O	O
leukemic	NN	O	B-cell_type
cell	NN	O	I-cell_type
in	NN	O	O
10	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
PML/RAR	NN	O	B-RNA
alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
still	NN	O	O
detectable	NN	O	O
despite	NN	O	O
clinical	NN	O	O
remission	NN	O	O
following	NN	O	O
ATRA	NN	O	O
treatment	NN	O	O
,	NN	O	O
but	NN	O	O
undetectable	NN	O	O
following	NN	O	O
consolidation	NN	O	O
with	NN	O	O
chemotherapy	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
technique	NN	O	O
is	NN	O	O
useful	NN	O	O
for	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
minimal	NN	O	O
residual	NN	O	O
disease	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
APL	NN	O	O
and	NN	O	O
that	NN	O	O
cytotoxic	NN	O	O
chemotherapy	NN	O	O
following	NN	O	O
ATRA	NN	O	O
therapy	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
elimination	NN	O	O
of	NN	O	O
APL	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Glucocorticoid-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
interleukin-2	NN	O	O
receptor	NN	O	O
alpha	NN	O	O
and	NN	O	O
-beta	NN	O	O
subunit	NN	O	O
expression	NN	O	O
by	NN	O	O
human	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
functions	NN	O	O
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
DM	NN	O	O
)	NN	O	O
on	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
and	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-2R	NN	O	B-protein
)	NN	O	O
generation	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
inhibited	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
by	NN	O	O
30	NN	O	O
%	NN	O	O
-88	NN	O	O
%	NN	O	O
,	NN	O	O
relative	NN	O	O
to	NN	O	O
its	NN	O	O
concentration	NN	O	O
within	NN	O	O
the	NN	O	O
cultures	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
DM	NN	O	O
on	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
(	NN	O	O
Tac	NN	O	B-protein
,	NN	O	O
p55	NN	O	B-protein
,	NN	O	O
CD25	NN	O	B-protein
)	NN	O	O
and	NN	O	O
beta	NN	O	B-DNA
(	NN	O	I-DNA
p75	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
examined	NN	O	O
next	NN	O	O
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
purified	NN	O	O
phytohemagglutinin	NN	O	B-protein
(	NN	O	O
PHA-p	NN	O	B-protein
)	NN	O	O
and	NN	O	O
4	NN	O	O
beta-phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
addition	NN	O	O
of	NN	O	O
DM	NN	O	O
to	NN	O	O
the	NN	O	O
cultures	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
60	NN	O	O
%	NN	O	O
reduction	NN	O	O
in	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
a	NN	O	O
30	NN	O	O
%	NN	O	O
reduction	NN	O	O
in	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
membrane	NN	O	O
expression	NN	O	O
compared	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
cultured	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
DM	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
membrane	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
M	NN	O	O
DM	NN	O	O
was	NN	O	O
partially	NN	O	O
reversible	NN	O	O
by	NN	O	O
recombinant	NN	O	B-protein
human	NN	O	I-protein
IL-2	NN	O	I-protein
(	NN	O	O
rhIL-2	NN	O	B-protein
)	NN	O	O
.	NN	O	O

By	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
DM	NN	O	O
caused	NN	O	O
a	NN	O	O
comparable	NN	O	O
decrease	NN	O	O
in	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
in	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
mRNA	NN	O	O
levels	NN	O	O
to	NN	O	O
membrane	NN	O	O
receptor	NN	O	O
expression	NN	O	O
in	NN	O	O
mitogen-stimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

By	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
assays	NN	O	O
,	NN	O	O
DM	NN	O	O
regulated	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
at	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
while	NN	O	O
transcription	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-DNA
beta	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
DM	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
DM	NN	O	O
on	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
transcription	NN	O	O
was	NN	O	O
examined	NN	O	O
by	NN	O	O
determining	NN	O	O
the	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
p50	NN	O	O
subunit	NN	O	O
of	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
kappa	NN	O	O
B	NN	O	O
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
stimulates	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
M	NN	O	O
DM	NN	O	O
increased	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
p50	NN	O	O
NF-kappa	NN	O	B-RNA
B	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
by	NN	O	O
four-fold	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
levels	NN	O	O
obtained	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
DM	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
increased	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
DM	NN	O	O
.	NN	O	O

In	NN	O	O
sum	NN	O	O
,	NN	O	O
DM	NN	O	O
regulates	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
membrane	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
by	NN	O	O
more	NN	O	O
than	NN	O	O
one	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
.	NN	O	O

-DOCSTART-	O

Alteration	NN	O	O
of	NN	O	O
structural	NN	O	O
order	NN	O	O
of	NN	O	O
human	NN	O	O
erythrocyte	NN	O	O
ghost	NN	O	O
membrane	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
antagonist	NN	O	O
RU	NN	O	O
486	NN	O	O
.	NN	O	O

High-dose	NN	O	O
pulse	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
successfully	NN	O	O
in	NN	O	O
the	NN	O	O
clinic	NN	O	O
in	NN	O	O
severe	NN	O	O
pathological	NN	O	O
conditions	NN	O	O
for	NN	O	O
about	NN	O	O
20	NN	O	O
years	NN	O	O
.	NN	O	O

The	NN	O	O
mode	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
action	NN	O	O
after	NN	O	O
administration	NN	O	O
of	NN	O	O
such	NN	O	O
megadoses	NN	O	O
is	NN	O	O
inexplicable	NN	O	O
up	NN	O	O
to	NN	O	O
now	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
supposed	NN	O	O
that	NN	O	O
some	NN	O	O
effects	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
membrane	NN	O	O
alterations	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
in-vitro	NN	O	O
experiments	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
of	NN	O	O
further	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
and	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
antagonist	NN	O	O
RU	NN	O	O
486	NN	O	O
,	NN	O	O
on	NN	O	O
structural	NN	O	O
order	NN	O	O
of	NN	O	O
human	NN	O	O
erythrocyte	NN	O	O
ghost	NN	O	O
membranes	NN	O	O
was	NN	O	O
investigated	NN	O	O
by	NN	O	O
determining	NN	O	O
the	NN	O	O
steady-state	NN	O	O
fluorescence	NN	O	O
anisotropy	NN	O	O
of	NN	O	O
diphenylhexatriene	NN	O	O
(	NN	O	O
DPH	NN	O	O
)	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
induce	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
in	NN	O	O
membrane	NN	O	O
structural	NN	O	O
order	NN	O	O
at	NN	O	O
concentrations	NN	O	O
of	NN	O	O
about	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
M	NN	O	O
in	NN	O	O
a	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
a	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
uptake	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
by	NN	O	O
the	NN	O	O
ghost	NN	O	O
membranes	NN	O	O
and	NN	O	O
the	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
structural	NN	O	O
order	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
glucocorticoids	NN	O	O
tested	NN	O	O
,	NN	O	O
methylprednisolone	NN	O	O
and	NN	O	O
corticosterone	NN	O	O
,	NN	O	O
were	NN	O	O
also	NN	O	O
effective	NN	O	O
at	NN	O	O
concentrations	NN	O	O
of	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
M	NN	O	O
or	NN	O	O
greater	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
membrane	NN	O	O
structural	NN	O	O
order	NN	O	O
with	NN	O	O
RU	NN	O	O
486	NN	O	O
up	NN	O	O
to	NN	O	O
a	NN	O	O
concentration	NN	O	O
of	NN	O	O
10	NN	O	O
(	NN	O	O
-4	NN	O	O
)	NN	O	O
M	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
simultaneous	NN	O	O
incubation	NN	O	O
of	NN	O	O
RU	NN	O	O
486	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
caused	NN	O	O
a	NN	O	O
distinct	NN	O	O
interference	NN	O	O
of	NN	O	O
RU	NN	O	O
486	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
glucocorticoid-induced	NN	O	O
membrane	NN	O	O
perturbation	NN	O	O
,	NN	O	O
the	NN	O	O
possibility	NN	O	O
to	NN	O	O
inhibit	NN	O	O
it	NN	O	O
by	NN	O	O
RU	NN	O	O
486	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
inactivity	NN	O	O
of	NN	O	O
the	NN	O	O
structurally	NN	O	O
related	NN	O	O
progesterone	NN	O	O
,	NN	O	O
refer	NN	O	O
to	NN	O	O
relatively	NN	O	O
specific	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
the	NN	O	O
membrane	NN	O	O
of	NN	O	O
erythrocyte	NN	O	B-cell_type
ghosts	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Mutation	NN	O	O
of	NN	O	O
tyrosines	NN	O	O
492/493	NN	O	O
in	NN	O	O
the	NN	O	O
kinase	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
ZAP-70	NN	O	B-protein
affects	NN	O	O
multiple	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

The	NN	O	O
protein-tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
ZAP-70	NN	O	B-protein
is	NN	O	O
implicated	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
the	NN	O	O
Src	NN	O	B-protein
kinase	NN	O	I-protein
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
in	NN	O	O
controlling	NN	O	O
the	NN	O	O
early	NN	O	O
steps	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
cascade	NN	O	O
.	NN	O	O

To	NN	O	O
help	NN	O	O
elucidate	NN	O	O
further	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
ZAP-70	NN	O	B-protein
regulates	NN	O	O
these	NN	O	O
initial	NN	O	O
events	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
a	NN	O	O
dominant-negative	NN	O	O
mutant	NN	O	O
approach	NN	O	O
.	NN	O	O

We	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
ZAP-70	NN	O	B-protein
mutated	NN	O	O
on	NN	O	O
Tyr-492	NN	O	O
and	NN	O	O
Tyr-493	NN	O	O
in	NN	O	O
the	NN	O	O
putative	NN	O	O
regulatory	NN	O	O
loop	NN	O	O
of	NN	O	O
its	NN	O	O
kinase	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

This	NN	O	O
mutant	NN	O	O
inhibited	NN	O	O
TCR	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
both	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
increase	NN	O	O
and	NN	O	O
Ras	NN	O	B-protein
-regulated	NN	O	O
activation	NN	O	O
of	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
TCR	NN	O	B-protein
-induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
pp36-38	NN	O	B-protein
,	NN	O	O
thought	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
upstream	NN	O	O
of	NN	O	O
these	NN	O	O
pathways	NN	O	O
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
reduced	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
wild-type	NN	O	B-protein
ZAP-70	NN	O	I-protein
induced	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
.	NN	O	O

The	NN	O	O
ZAP-70	NN	O	B-protein
mutant	NN	O	O
studied	NN	O	O
here	NN	O	O
could	NN	O	O
be	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
when	NN	O	O
associated	NN	O	O
to	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
zeta	NN	O	I-protein
chain	NN	O	I-protein
and	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
bind	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

This	NN	O	O
result	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
Tyr-492	NN	O	O
and	NN	O	O
Tyr-493	NN	O	O
are	NN	O	O
not	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
Src	NN	O	O
homology	NN	O	O
domain	NN	O	O
2-mediated	NN	O	O
association	NN	O	O
of	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
with	NN	O	O
ZAP-70	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
data	NN	O	O
are	NN	O	O
most	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
recruitment	NN	O	O
to	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
allows	NN	O	O
ZAP-70	NN	O	B-protein
autophosphorylation	NN	O	O
and	NN	O	O
binding	NN	O	O
to	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
phosphorylates	NN	O	O
Tyr-492	NN	O	O
and/or	NN	O	O
Tyr-493	NN	O	O
with	NN	O	O
consequent	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
ZAP-70	NN	O	B-protein
kinase	NN	O	O
activity	NN	O	O
.	NN	O	O

ZAP-70	NN	O	B-protein
will	NN	O	O
then	NN	O	O
be	NN	O	O
able	NN	O	O
to	NN	O	O
effectively	NN	O	O
control	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
its	NN	O	O
substrates	NN	O	O
and	NN	O	O
lead	NN	O	O
to	NN	O	O
gene	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
on	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
steroid-sensitive	NN	O	O
and	NN	O	O
steroid-resistant	NN	O	O
asthma	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Glucocorticoids	NN	O	O
are	NN	O	O
important	NN	O	O
medications	NN	O	O
used	NN	O	O
to	NN	O	O
control	NN	O	O
the	NN	O	O
airway	NN	O	O
inflammation	NN	O	O
associated	NN	O	O
with	NN	O	O
asthma	NN	O	O
.	NN	O	O

Synthetic	NN	O	O
glucocorticoids	NN	O	O
vary	NN	O	O
in	NN	O	O
their	NN	O	O
binding	NN	O	O
affinity	NN	O	O
for	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
(	NN	O	O
GCR	NN	O	O
)	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
We	NN	O	O
compared	NN	O	O
hydrocortisone	NN	O	O
,	NN	O	O
beclomethasone	NN	O	O
dipropionate	NN	O	O
,	NN	O	O
triamcinolone	NN	O	O
acetonide	NN	O	O
,	NN	O	O
flunisolide	NN	O	O
,	NN	O	O
and	NN	O	O
budesonide	NN	O	O
with	NN	O	O
regard	NN	O	O
to	NN	O	O
their	NN	O	O
capacity	NN	O	O
to	NN	O	O
inhibit	NN	O	O
phytohemagglutinin	NN	O	B-protein
-induced	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
from	NN	O	O
six	NN	O	O
patients	NN	O	O
with	NN	O	O
steroid-sensitive	NN	O	O
asthma	NN	O	O
and	NN	O	O
seven	NN	O	O
patients	NN	O	O
with	NN	O	O
steroid-resistant	NN	O	O
asthma	NN	O	O
.	NN	O	O

Peripheral	NN	O	O
blood	NN	O	O
mononuclear	NN	O	O
cell	NN	O	O
GCR	NN	O	B-protein
binding	NN	O	O
affinities	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
budesonide	NN	O	O
were	NN	O	O
also	NN	O	O
determined	NN	O	O
for	NN	O	O
both	NN	O	O
patient	NN	O	O
groups	NN	O	O
by	NN	O	O
using	NN	O	O
a	NN	O	O
radioligand	NN	O	O
binding	NN	O	O
assay	NN	O	O
and	NN	O	O
Scatchard	NN	O	O
analysis	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Dose-dependent	NN	O	O
inhibition	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
for	NN	O	O
all	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
both	NN	O	O
patient	NN	O	O
groups	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
steroid-resistant	NN	O	O
group	NN	O	O
requiring	NN	O	O
approximately	NN	O	O
2	NN	O	O
log-fold	NN	O	O
more	NN	O	O
glucocorticoids	NN	O	O
for	NN	O	O
an	NN	O	O
equivalent	NN	O	O
degree	NN	O	O
of	NN	O	O
inhibition	NN	O	O
.	NN	O	O

The	NN	O	O
mean	NN	O	O
concentrations	NN	O	O
necessary	NN	O	O
to	NN	O	O
cause	NN	O	O
50	NN	O	O
%	NN	O	O
inhibition	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
(	NN	O	O
IC50s	NN	O	O
)	NN	O	O
for	NN	O	O
the	NN	O	O
steroid-sensitive	NN	O	O
group	NN	O	O
ranged	NN	O	O
from	NN	O	O
2	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
mol/L	NN	O	O
for	NN	O	O
budesonide	NN	O	O
to	NN	O	O
7	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
mol/L	NN	O	O
for	NN	O	O
hydrocortisone	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
mean	NN	O	O
IC50s	NN	O	O
for	NN	O	O
the	NN	O	O
steroid-resistant	NN	O	O
group	NN	O	O
ranged	NN	O	O
from	NN	O	O
approximately	NN	O	O
2	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
mol/L	NN	O	O
for	NN	O	O
budesonide	NN	O	O
to	NN	O	O
greater	NN	O	O
than	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
mol/L	NN	O	O
for	NN	O	O
hydrocortisone	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
correlation	NN	O	O
was	NN	O	O
noted	NN	O	O
between	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
inhibition	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
(	NN	O	O
IC50	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
to	NN	O	O
the	NN	O	O
GCR	NN	O	B-protein
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
steroid-resistant	NN	O	O
asthma	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
reduced	NN	O	O
GCR	NN	O	B-protein
binding	NN	O	O
affinity	NN	O	O
.	NN	O	O

The	NN	O	O
GCR	NN	O	B-protein
binding	NN	O	O
affinity	NN	O	O
for	NN	O	O
budesonide	NN	O	O
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
in	NN	O	O
both	NN	O	O
groups	NN	O	O
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
lower	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
)	NN	O	O
than	NN	O	O
that	NN	O	O
obtained	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
such	NN	O	O
as	NN	O	O
budesonide	NN	O	O
,	NN	O	O
by	NN	O	O
virtue	NN	O	O
of	NN	O	O
their	NN	O	O
high	NN	O	O
GCR	NN	O	B-protein
binding	NN	O	O
affinities	NN	O	O
and	NN	O	O
greater	NN	O	O
ability	NN	O	O
to	NN	O	O
suppress	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
,	NN	O	O
may	NN	O	O
therefore	NN	O	O
be	NN	O	O
beneficial	NN	O	O
in	NN	O	O
the	NN	O	O
management	NN	O	O
of	NN	O	O
difficult-to-control	NN	O	O
asthma	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
cyclin-dependent	NN	O	I-DNA
kinase	NN	O	I-DNA
inhibitor	NN	O	I-DNA
gene	NN	O	I-DNA
p27Kip1	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
cyclin-dependent	NN	O	B-DNA
kinase	NN	O	I-DNA
inhibitor	NN	O	I-DNA
p27Kip1	NN	O	B-DNA
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
cell-cycle	NN	O	O
progression	NN	O	O
.	NN	O	O

p27Kip1	NN	O	B-DNA
directly	NN	O	O
inhibits	NN	O	O
the	NN	O	O
catalytic	NN	O	O
activity	NN	O	O
of	NN	O	O
cyclin/cdks	NN	O	B-protein
(	NN	O	I-protein
cyclin-dependent	NN	O	I-protein
kinase	NN	O	I-protein
)	NN	O	I-protein
complexes	NN	O	I-protein
and/or	NN	O	O
interferes	NN	O	O
physically	NN	O	O
with	NN	O	O
cyclin/cdks	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
CAK	NN	O	B-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
level	NN	O	O
of	NN	O	O
p27Kip1	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
maximal	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
Go	NN	O	I-cell_type
T-cells	NN	O	I-cell_type
and	NN	O	O
rapidly	NN	O	O
declined	NN	O	O
following	NN	O	O
anti-CD3	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
cloning	NN	O	O
of	NN	O	O
p27Kip1	NN	O	B-DNA
gene	NN	O	I-DNA
from	NN	O	O
murine	NN	O	B-DNA
genomic	NN	O	I-DNA
DNA	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
of	NN	O	O
the	NN	O	O
p27Kip1	NN	O	B-DNA
gene	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
consists	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
exons	NN	O	O
and	NN	O	O
spans	NN	O	O
more	NN	O	O
than	NN	O	O
5.6	NN	O	B-DNA
kb	NN	O	I-DNA
of	NN	O	I-DNA
DNA	NN	O	I-DNA
.	NN	O	O

Primer	NN	O	O
extension	NN	O	O
and	NN	O	O
nuclease	NN	O	O
S1	NN	O	O
protection	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
two	NN	O	O
major	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
lacked	NN	O	O
a	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
but	NN	O	O
contained	NN	O	O
potential	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
factors	NN	O	I-protein
including	NN	O	O
two	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
CRE	NN	O	B-protein
,	NN	O	O
Myb	NN	O	B-protein
and	NN	O	O
NFkB	NN	O	B-protein
located	NN	O	O
at	NN	O	O
positions	NN	O	B-DNA
-153	NN	O	I-DNA
,	NN	O	I-DNA
-178	NN	O	I-DNA
,	NN	O	I-DNA
-286	NN	O	I-DNA
,	NN	O	I-DNA
-875	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
-1011	NN	O	I-DNA
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

To	NN	O	O
analyze	NN	O	O
the	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
controlling	NN	O	O
p27Kip1	NN	O	B-DNA
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
characterized	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
from	NN	O	O
nt	NN	O	B-DNA
-1609	NN	O	I-DNA
to	NN	O	I-DNA
+178	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
-326	NN	O	B-DNA
to	NN	O	I-DNA
-615	NN	O	I-DNA
region	NN	O	I-DNA
contained	NN	O	O
positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
NF-ATp	NN	O	I-protein
is	NN	O	O
a	NN	O	O
hallmark	NN	O	O
of	NN	O	O
unstimulated	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
B-CLL	NN	O	O
patients	NN	O	O
.	NN	O	O

B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukaemia	NN	O	O
(	NN	O	O
CLL	NN	O	O
)	NN	O	O
were	NN	O	O
analysed	NN	O	O
for	NN	O	O
the	NN	O	O
nuclear	NN	O	O
presence	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
which	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
control	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
following	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
were	NN	O	O
studied	NN	O	O
:	NN	O	O
the	NN	O	O
Octamer	NN	O	B-protein
factors	NN	O	I-protein
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
factor	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
NF-AT	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
in	NN	O	O
particular	NN	O	O
NF-ATp	NN	O	B-protein
,	NN	O	O
and	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Rel/NF-kB	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
constitutive	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-ATp	NN	O	B-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
growing	NN	O	O
family	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
is	NN	O	O
a	NN	O	O
hallmark	NN	O	O
of	NN	O	O
nonstimulated	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
CLL	NN	O	O
patients	NN	O	O
that	NN	O	O
distinguishes	NN	O	O
B-CLL	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
'normal	NN	O	B-cell_type
'	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Constitutive	NN	O	O
nuclear	NN	O	O
appearance	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
for	NN	O	O
NF-kB2/p52	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
binding	NN	O	O
of	NN	O	O
further	NN	O	O
factor	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
DNA	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
JunD	NN	O	B-protein
,	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
FosB	NN	O	B-protein
,	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
several	NN	O	O
patients	NN	O	O
whereas	NN	O	O
the	NN	O	O
localisation	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
other	NN	O	O
factors	NN	O	O
such	NN	O	O
as	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
RelA/p65	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
was	NN	O	O
unaltered	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
remarkable	NN	O	O
that	NN	O	O
in	NN	O	O
B-CLL	NN	O	B-cell_type
cells	NN	O	I-cell_type
the	NN	O	O
nuclear	NN	O	O
appearance	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
specific	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
is	NN	O	O
dramatically	NN	O	O
affected	NN	O	O
whereas	NN	O	O
other	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
factor	NN	O	B-protein
family	NN	O	I-protein
remained	NN	O	O
unaltered	NN	O	O
in	NN	O	O
these	NN	O	O
leukemic	NN	O	O
cells	NN	O	O
.	NN	O	O

It	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
shown	NN	O	O
which	NN	O	O
molecular	NN	O	O
events	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
specific	NN	O	O
'pre-activation	NN	O	O
'	NN	O	O
,	NN	O	O
i.e	NN	O	O
.	NN	O	O

constitutive	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
of	NN	O	O
these	NN	O	O
members	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
NF-kB	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
factor	NN	O	I-protein
families	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Naive	NN	O	B-cell_type
(	NN	O	I-cell_type
CD45RA+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
are	NN	O	O
more	NN	O	O
sensitive	NN	O	O
to	NN	O	O
oxidative	NN	O	O
stress-induced	NN	O	O
signals	NN	O	O
than	NN	O	O
memory	NN	O	B-cell_type
(	NN	O	I-cell_type
CD45RO+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Formation	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
(	NN	O	O
ROI	NN	O	O
)	NN	O	O
after	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
activation	NN	O	O
signal	NN	O	O
for	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
e.g.	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
its	NN	O	O
receptor	NN	O	B-protein
are	NN	O	O
induced	NN	O	O
.	NN	O	O

These	NN	O	O
ROI-induced	NN	O	O
effects	NN	O	O
can	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
large	NN	O	O
extent	NN	O	O
,	NN	O	O
be	NN	O	O
attributed	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

Now	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
whether	NN	O	O
naive	NN	O	B-cell_type
and	NN	O	I-cell_type
memory	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
differ	NN	O	O
in	NN	O	O
their	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
ROI-mediated	NN	O	O
signals	NN	O	O
.	NN	O	O

When	NN	O	O
CD45RA+	NN	O	B-cell_type
(	NN	O	I-cell_type
naive	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	O
CD45RO+	NN	O	B-cell_type
(	NN	O	I-cell_type
memory	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
directly	NN	O	O
stimulated	NN	O	O
with	NN	O	O
H2O2	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
was	NN	O	O
stronger	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
cells	NN	O	I-cell_type
than	NN	O	O
in	NN	O	O
memory	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
it	NN	O	O
could	NN	O	O
be	NN	O	O
induced	NN	O	O
with	NN	O	O
lower	NN	O	O
doses	NN	O	O
.	NN	O	O

The	NN	O	O
composition	NN	O	O
of	NN	O	O
the	NN	O	O
induced	NN	O	O
nuclear	NN	O	B-protein
NF-kappaB	NN	O	I-protein
(	NN	O	O
levels	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
RelA	NN	O	B-protein
proteins	NN	O	I-protein
)	NN	O	O
was	NN	O	O
similar	NN	O	O
in	NN	O	O
these	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

The	NN	O	O
magnitude	NN	O	O
and	NN	O	O
kinetics	NN	O	O
of	NN	O	O
intracellular	NN	O	O
ROI	NN	O	O
were	NN	O	O
similar	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
there	NN	O	O
were	NN	O	O
no	NN	O	O
differences	NN	O	O
in	NN	O	O
ROI-forming	NN	O	O
mechanisms	NN	O	O
or	NN	O	O
antioxidative	NN	O	O
capacities	NN	O	O
.	NN	O	O

The	NN	O	O
probable	NN	O	O
regulatory	NN	O	O
point	NN	O	O
was	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
IkappaB	NN	O	B-protein
inhibitor	NN	O	O
:	NN	O	O
in	NN	O	O
CD45RA+	NN	O	O
cells	NN	O	O
,	NN	O	O
H2O2	NN	O	O
caused	NN	O	O
a	NN	O	O
more	NN	O	O
profound	NN	O	O
depression	NN	O	O
in	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
representing	NN	O	O
different	NN	O	O
activation	NN	O	O
and/or	NN	O	O
differentiation	NN	O	O
stages	NN	O	O
can	NN	O	O
be	NN	O	O
differentially	NN	O	O
responsive	NN	O	O
to	NN	O	O
ROI-mediated	NN	O	O
signals	NN	O	O
.	NN	O	O

-DOCSTART-	O

Rapid	NN	O	O
shuttling	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
in	NN	O	O
discrimination	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
signals	NN	O	O
and	NN	O	O
immunosuppression	NN	O	O
.	NN	O	O

Cells	NN	O	O
need	NN	O	O
to	NN	O	O
distinguish	NN	O	O
between	NN	O	O
transient	NN	O	O
Ca2+	NN	O	O
signals	NN	O	O
that	NN	O	O
induce	NN	O	O
events	NN	O	O
such	NN	O	O
as	NN	O	O
muscle	NN	O	O
contraction	NN	O	O
,	NN	O	O
secretion	NN	O	O
,	NN	O	O
adhesion	NN	O	O
and	NN	O	O
synaptic	NN	O	O
transmission	NN	O	O
,	NN	O	O
and	NN	O	O
sustained	NN	O	O
Ca2+	NN	O	O
signals	NN	O	O
that	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
latter	NN	O	O
class	NN	O	O
of	NN	O	O
events	NN	O	O
is	NN	O	O
blocked	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
by	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
drugs	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
,	NN	O	O
which	NN	O	O
inhibit	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
Ca2+-activated	NN	O	B-protein
serine/threonine	NN	O	I-protein
phosphatase	NN	O	I-protein
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
nuclear	NN	O	O
import	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
sustained	NN	O	O
high	NN	O	O
concentrations	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
transient	NN	O	O
pulses	NN	O	O
,	NN	O	O
are	NN	O	O
required	NN	O	O
to	NN	O	O
maintain	NN	O	O
NF-AT	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
where	NN	O	O
they	NN	O	O
participate	NN	O	O
in	NN	O	O
Ca2+-dependent	NN	O	O
induction	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
required	NN	O	O
for	NN	O	O
lymphocyte	NN	O	B-cell_type
activation	NN	O	O
and	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
overexpression	NN	O	O
and	NN	O	O
constitutive	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Jun	NN	O	B-protein
,	NN	O	I-protein
Fos	NN	O	I-protein
,	NN	O	I-protein
NF-kappaB	NN	O	I-protein
,	NN	O	I-protein
Oct	NN	O	I-protein
or	NN	O	I-protein
Ets	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
,	NN	O	O
renders	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
resistant	NN	O	O
to	NN	O	O
FK506	NN	O	O
and	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

Thus	NN	O	O
a	NN	O	O
primary	NN	O	O
effect	NN	O	O
of	NN	O	O
these	NN	O	O
immunosuppressive	NN	O	O
reagents	NN	O	O
is	NN	O	O
to	NN	O	O
control	NN	O	O
the	NN	O	O
subcellular	NN	O	O
localization	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Autoregulation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transactivator	NN	O	I-protein
RelA	NN	O	B-protein
(	NN	O	O
p65	NN	O	B-protein
)	NN	O	O
by	NN	O	O
multiple	NN	O	O
cytoplasmic	NN	O	B-protein
inhibitors	NN	O	I-protein
containing	NN	O	O
ankyrin	NN	O	B-protein
motifs	NN	O	I-protein
.	NN	O	O

RelA	NN	O	B-protein
(	NN	O	O
p65	NN	O	B-protein
)	NN	O	O
functions	NN	O	O
as	NN	O	O
the	NN	O	O
critical	NN	O	O
transactivating	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
heterodimeric	NN	O	B-protein
p50-p65	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
complex	NN	O	I-protein
and	NN	O	O
contains	NN	O	O
a	NN	O	O
high-affinity	NN	O	B-protein
binding	NN	O	I-protein
site	NN	O	I-protein
for	NN	O	O
its	NN	O	O
cytoplasmic	NN	O	B-protein
inhibitor	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

After	NN	O	O
cellular	NN	O	O
activation	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
is	NN	O	O
rapidly	NN	O	O
degraded	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
the	NN	O	O
induced	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
-induced	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
preceded	NN	O	O
by	NN	O	O
its	NN	O	O
rapid	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
these	NN	O	O
effects	NN	O	O
on	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
result	NN	O	O
in	NN	O	O
nuclear	NN	O	O
mobilization	NN	O	O
of	NN	O	O
only	NN	O	O
a	NN	O	O
fraction	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	O
cytoplasmic	NN	O	O
pool	NN	O	O
of	NN	O	O
RelA	NN	O	B-protein
.	NN	O	O

Subsequent	NN	O	O
studies	NN	O	O
have	NN	O	O
revealed	NN	O	O
that	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
cytoplasmic	NN	O	B-protein
RelA	NN	O	I-protein
is	NN	O	O
stably	NN	O	O
associated	NN	O	O
not	NN	O	O
only	NN	O	O
with	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
but	NN	O	O
also	NN	O	O
with	NN	O	O
other	NN	O	O
ankyrin	NN	O	B-protein
motif-rich	NN	O	I-protein
proteins	NN	O	I-protein
including	NN	O	O
the	NN	O	O
products	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B2	NN	O	I-DNA
(	NN	O	I-DNA
p100	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B1	NN	O	I-DNA
(	NN	O	I-DNA
p105	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
RelA	NN	O	B-protein
-I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
RelA-p100	NN	O	B-protein
cytoplasmic	NN	O	I-protein
complexes	NN	O	I-protein
are	NN	O	O
not	NN	O	O
dissociated	NN	O	O
following	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
activation	NN	O	O
;	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
p100	NN	O	B-protein
functions	NN	O	O
as	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
RelA	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
in	NN	O	O
vivo	NN	O	O
;	NN	O	O
(	NN	O	O
iv	NN	O	O
)	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
RelA	NN	O	B-protein
and	NN	O	O
p100	NN	O	B-protein
involves	NN	O	O
the	NN	O	O
conserved	NN	O	B-protein
Rel	NN	O	I-protein
homology	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
both	NN	O	O
proteins	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
localization	NN	O	I-protein
signal	NN	O	I-protein
of	NN	O	O
RelA	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
binding	NN	O	O
;	NN	O	O
(	NN	O	O
v	NN	O	O
)	NN	O	O
p100	NN	O	B-protein
inhibition	NN	O	O
of	NN	O	O
RelA	NN	O	B-protein
function	NN	O	O
requires	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
ankyrin	NN	O	I-protein
motif	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
which	NN	O	O
mediates	NN	O	O
cytoplasmic	NN	O	O
retention	NN	O	O
of	NN	O	O
RelA	NN	O	B-protein
;	NN	O	O
and	NN	O	O
(	NN	O	O
vi	NN	O	O
)	NN	O	O
as	NN	O	O
observed	NN	O	O
with	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
nuclear	NN	O	O
RelA	NN	O	B-protein
stimulates	NN	O	O
p100	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
thus	NN	O	O
reveal	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
second	NN	O	O
inducible	NN	O	O
autoregulated	NN	O	O
inhibitory	NN	O	O
pathway	NN	O	O
that	NN	O	O
helps	NN	O	O
ensure	NN	O	O
the	NN	O	O
rapid	NN	O	O
but	NN	O	O
transient	NN	O	O
action	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
CD45	NN	O	B-protein
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Implications	NN	O	O
for	NN	O	O
replication	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

Increased	NN	O	O
levels	NN	O	O
of	NN	O	O
replication	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	O
type	NN	O	O
1	NN	O	O
are	NN	O	O
observed	NN	O	O
after	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
infected	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
anti-	NN	O	O
CD45	NN	O	B-protein
antibodies	NN	O	O
inhibit	NN	O	O
these	NN	O	O
effects	NN	O	O
in	NN	O	O
cells	NN	O	O
from	NN	O	O
infected	NN	O	O
individuals	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
interrelationships	NN	O	O
between	NN	O	O
CD45	NN	O	B-protein
and	NN	O	O
HIV-1	NN	O	O
further	NN	O	O
.	NN	O	O

We	NN	O	O
measured	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
antibodies	NN	O	O
against	NN	O	O
CD45	NN	O	B-protein
and	NN	O	O
in	NN	O	O
those	NN	O	O
that	NN	O	O
lacked	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CD45	NN	O	B-protein
on	NN	O	O
their	NN	O	O
surfaces	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
anti-CD45	NN	O	B-protein
antibodies	NN	O	I-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
basal	NN	O	O
but	NN	O	O
decreased	NN	O	O
activated	NN	O	O
levels	NN	O	O
of	NN	O	O
expression	NN	O	O
from	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

Second	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
lack	NN	O	O
CD45	NN	O	B-protein
and	NN	O	O
can	NN	O	O
not	NN	O	O
signal	NN	O	O
via	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
,	NN	O	O
supported	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
viral	NN	O	O
replication	NN	O	O
and	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
active	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
CD45	NN	O	B-protein
-T	NN	O	O
cells	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
infected	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
displayed	NN	O	O
lower	NN	O	O
levels	NN	O	O
of	NN	O	O
CD45	NN	O	B-protein
on	NN	O	O
their	NN	O	O
surfaces	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
CD45	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
active	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
physiology	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
the	NN	O	O
replication	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

-DOCSTART-	O

Calcineurin	NN	O	B-protein
potentiates	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
granulocyte-macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
conserved	NN	O	B-DNA
lymphokine	NN	O	I-DNA
element	NN	O	I-DNA
0	NN	O	I-DNA
.	NN	O	O

Granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
are	NN	O	O
produced	NN	O	O
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phorbol-12-myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
(	NN	O	O
A23187	NN	O	O
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
immunosuppressive	NN	O	O
drugs	NN	O	O
such	NN	O	O
as	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
and	NN	O	O
FK506	NN	O	O
.	NN	O	O

Earlier	NN	O	O
studies	NN	O	O
on	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
showed	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
(	NN	O	O
CN	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
Ca2+/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
phosphatase	NN	O	I-protein
,	NN	O	O
can	NN	O	O
stimulate	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
through	NN	O	O
the	NN	O	O
NF-AT-binding	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
obtained	NN	O	O
evidence	NN	O	O
that	NN	O	O
transfection	NN	O	O
of	NN	O	O
the	NN	O	O
cDNAs	NN	O	B-DNA
for	NN	O	O
CN	NN	O	B-protein
A	NN	O	I-protein
(	NN	O	I-protein
catalytic	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
CN	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
regulatory	NN	O	I-protein
)	NN	O	I-protein
subunits	NN	O	I-protein
also	NN	O	O
augments	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
recovers	NN	O	O
the	NN	O	O
transcription	NN	O	O
inhibited	NN	O	O
by	NN	O	O
CsA	NN	O	O
.	NN	O	O

The	NN	O	O
constitutively	NN	O	O
active	NN	O	O
type	NN	O	O
of	NN	O	O
the	NN	O	O
CN	NN	O	B-protein
A	NN	O	I-protein
subunit	NN	O	I-protein
,	NN	O	O
which	NN	O	O
lacks	NN	O	O
the	NN	O	O
auto-inhibitory	NN	O	B-protein
and	NN	O	O
calmodulin-binding	NN	O	B-protein
domains	NN	O	I-protein
,	NN	O	O
acts	NN	O	O
in	NN	O	O
synergy	NN	O	O
with	NN	O	O
PMA	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
also	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
active	NN	O	O
CN	NN	O	B-protein
partially	NN	O	O
replaces	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
in	NN	O	O
synergy	NN	O	O
with	NN	O	O
PMA	NN	O	O
to	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
endogenous	NN	O	O
GM-CSF	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

By	NN	O	O
multimerizing	NN	O	O
the	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
target	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
CN	NN	O	B-protein
action	NN	O	O
is	NN	O	O
the	NN	O	O
conserved	NN	O	B-DNA
lymphokine	NN	O	I-DNA
element	NN	O	I-DNA
0	NN	O	I-DNA
(	NN	O	O
CLE0	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
located	NN	O	O
at	NN	O	O
positions	NN	O	O
between	NN	O	O
-54	NN	O	O
and	NN	O	O
-40	NN	O	O
.	NN	O	O

Mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
CLE0	NN	O	B-DNA
sequence	NN	O	O
has	NN	O	O
an	NN	O	O
AP1-binding	NN	O	B-DNA
site	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
NF-AT-like	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
termed	NN	O	O
NF-CLE0	NN	O	B-protein
gamma	NN	O	I-protein
.	NN	O	O

NF-	NN	O	O
CLE0	NN	O	B-DNA
gamma	NN	O	O
binding	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
PMA/A23187	NN	O	O
and	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
CsA	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
CN	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
coordinated	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
and	NN	O	I-DNA
IL-2	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
the	NN	O	O
CLE0	NN	O	B-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
functional	NN	O	O
analogue	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT-binding	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
mediates	NN	O	O
signals	NN	O	O
downstream	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Alternate	NN	O	O
immune	NN	O	O
system	NN	O	O
targets	NN	O	O
for	NN	O	O
TCDD	NN	O	O
:	NN	O	O
lymphocyte	NN	O	B-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
extrathymic	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
development	NN	O	O
.	NN	O	O

We	NN	O	O
here	NN	O	O
summarize	NN	O	O
evidence	NN	O	O
that	NN	O	O
thymic	NN	O	O
atrophy	NN	O	O
induced	NN	O	O
by	NN	O	O
2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8-tetrachlorodibenzo-p-dioxin	NN	O	O
(	NN	O	O
TCDD	NN	O	O
)	NN	O	O
can	NN	O	O
be	NN	O	O
mediated	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
damage	NN	O	O
to	NN	O	O
extrathymic	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
precursors	NN	O	I-cell_type
in	NN	O	O
bone	NN	O	O
marrow	NN	O	O
and	NN	O	O
fetal	NN	O	O
liver	NN	O	O
.	NN	O	O

This	NN	O	O
atrophy	NN	O	O
induction	NN	O	O
does	NN	O	O
not	NN	O	O
involve	NN	O	O
apoptotic	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
affected	NN	O	O
by	NN	O	O
the	NN	O	O
bcl-2	NN	O	B-DNA
proto-oncogene	NN	O	I-DNA
.	NN	O	O

TCDD	NN	O	O
mediates	NN	O	O
atrophy	NN	O	O
induction	NN	O	O
through	NN	O	O
its	NN	O	O
specific	NN	O	O
receptor	NN	O	O
(	NN	O	O
the	NN	O	O
AhR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
not	NN	O	O
through	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
estrogen	NN	O	O
receptor	NN	O	O
.	NN	O	O

Both	NN	O	O
TCDD	NN	O	O
and	NN	O	O
estradiol	NN	O	O
induce	NN	O	O
extrathymic	NN	O	O
T-cell	NN	O	O
differentiation	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
.	NN	O	O

These	NN	O	O
extrathymic	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
populations	NN	O	I-cell_type
include	NN	O	O
cells	NN	O	O
expressing	NN	O	O
elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
V	NN	O	B-protein
beta	NN	O	I-protein
T-cell	NN	O	I-protein
receptors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
normally	NN	O	O
deleted	NN	O	O
in	NN	O	O
thymic	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
Bcd	NN	O	B-DNA
,	NN	O	O
a	NN	O	O
novel	NN	O	O
proto-oncogene	NN	O	B-DNA
expressed	NN	O	O
in	NN	O	O
B-cells	NN	O	B-cell_type
.	NN	O	O

A	NN	O	O
novel	NN	O	B-cell_type
B-cell	NN	O	I-cell_type
derived	NN	O	O
(	NN	O	O
Bcd	NN	O	B-DNA
)	NN	O	O
oncogene	NN	O	B-DNA
has	NN	O	O
been	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
lymphocytes	NN	O	O
of	NN	O	O
one	NN	O	O
B-cell	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
B-CLL	NN	O	O
)	NN	O	O
patient	NN	O	O
using	NN	O	O
DNA	NN	O	O
transfer	NN	O	O
and	NN	O	O
a	NN	O	O
mouse	NN	O	O
tumorigenicity	NN	O	O
assay	NN	O	O
.	NN	O	O

The	NN	O	O
Bcd	NN	O	B-DNA
proto-oncogene	NN	O	B-DNA
was	NN	O	O
activated	NN	O	O
by	NN	O	O
a	NN	O	O
truncation	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
UTR	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
predicts	NN	O	O
for	NN	O	O
two	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frames	NN	O	I-DNA
(	NN	O	O
ORFs	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

ORF1	NN	O	B-DNA
consists	NN	O	O
of	NN	O	O
240	NN	O	O
bp	NN	O	O
that	NN	O	O
would	NN	O	O
encode	NN	O	O
80	NN	O	O
amino	NN	O	O
acids	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
ORF2	NN	O	I-DNA
consists	NN	O	O
of	NN	O	O
648	NN	O	O
bp	NN	O	O
capable	NN	O	O
of	NN	O	O
coding	NN	O	O
for	NN	O	O
216	NN	O	O
amino	NN	O	O
acids	NN	O	O
.	NN	O	O

Predicted	NN	O	B-protein
peptide	NN	O	I-protein
sequence	NN	O	I-protein
of	NN	O	O
ORF2	NN	O	B-DNA
contained	NN	O	O
a	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
domain	NN	O	I-protein
which	NN	O	O
showed	NN	O	O
significant	NN	O	O
homology	NN	O	O
to	NN	O	O
GC	NN	O	B-protein
box	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
BTEB2	NN	O	B-protein
and	NN	O	O
SP1	NN	O	B-protein
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
an	NN	O	O
expression	NN	O	B-DNA
vector	NN	O	I-DNA
containing	NN	O	O
ORF2	NN	O	B-DNA
but	NN	O	O
not	NN	O	O
full	NN	O	B-DNA
length	NN	O	I-DNA
cDNA	NN	O	I-DNA
was	NN	O	O
able	NN	O	O
to	NN	O	O
transform	NN	O	O
NIH3T3	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
induce	NN	O	O
tumors	NN	O	O
in	NN	O	O
nude	NN	O	O
mice	NN	O	O
.	NN	O	O

Bcd	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcripts	NN	O	I-RNA
of	NN	O	O
<	NN	O	O
or	NN	O	O
=	NN	O	O
2.6	NN	O	O
kb	NN	O	O
were	NN	O	O
selectively	NN	O	O
expressed	NN	O	O
in	NN	O	O
PBL	NN	O	B-cell_line
and	NN	O	O
testis	NN	O	O
of	NN	O	O
healthy	NN	O	O
individuals	NN	O	O
.	NN	O	O

Within	NN	O	O
the	NN	O	O
PBL	NN	O	B-cell_type
,	NN	O	O
Bcd	NN	O	B-DNA
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
CD19+	NN	O	B-cell_type
B-cells	NN	O	I-cell_type
and	NN	O	O
absent	NN	O	O
from	NN	O	O
CD14+	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
T-cells	NN	O	B-cell_type
.	NN	O	O

Bcd	NN	O	B-DNA
transcripts	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
all	NN	O	O
normal	NN	O	O
PBL	NN	O	B-cell_type
samples	NN	O	O
tested	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
several	NN	O	O
malignant	NN	O	B-cell_line
human	NN	O	I-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
not	NN	O	O
in	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
B-cells	NN	O	B-cell_type
from	NN	O	O
B-CLL	NN	O	O
patients	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
stimulation	NN	O	O
of	NN	O	O
B-cells	NN	O	B-cell_type
from	NN	O	O
B-CLL	NN	O	O
patients	NN	O	O
under	NN	O	O
conditions	NN	O	O
which	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
into	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
associated	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
Bcd	NN	O	B-DNA
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
Bcd	NN	O	B-DNA
gene	NN	O	I-DNA
may	NN	O	O
therefore	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
B-cell	NN	O	O
growth	NN	O	O
and	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	B-DNA
SP-1	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin-1	NN	O	I-DNA
beta	NN	O	I-DNA
promoter	NN	O	I-DNA
confers	NN	O	O
preferential	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
keratinocytes	NN	O	B-cell_type
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
interleukin-1beta	NN	O	B-protein
(	NN	O	O
IL-1beta	NN	O	B-protein
)	NN	O	O
in	NN	O	O
non-monocytic	NN	O	O
cells	NN	O	O
,	NN	O	O
we	NN	O	O
constructed	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
reporter	NN	O	O
plasmids	NN	O	O
with	NN	O	O
the	NN	O	O
bacterial	NN	O	O
chloramphenicol	NN	O	O
acetyltransferase	NN	O	O
gene	NN	O	O
linked	NN	O	O
to	NN	O	O
various	NN	O	O
parts	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-1beta	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
performed	NN	O	O
transient	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
promoter	NN	O	O
segment	NN	O	O
that	NN	O	O
activates	NN	O	O
transcription	NN	O	O
most	NN	O	O
efficiently	NN	O	O
in	NN	O	O
keratinocytes	NN	O	B-cell_type
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
with	NN	O	O
a	NN	O	O
43-mer	NN	O	B-DNA
oligonucleotide	NN	O	I-DNA
derived	NN	O	O
from	NN	O	O
the	NN	O	O
functionally	NN	O	O
identified	NN	O	O
cis-acting	NN	O	B-DNA
element	NN	O	I-DNA
revealed	NN	O	O
specific	NN	O	O
complexes	NN	O	O
.	NN	O	O

By	NN	O	O
competition	NN	O	O
analysis	NN	O	O
with	NN	O	O
transcription	NN	O	B-DNA
factor	NN	O	I-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
oligonucleotides	NN	O	I-DNA
and	NN	O	O
by	NN	O	O
immunosupershift	NN	O	O
,	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
SP-1	NN	O	I-protein
or	NN	O	O
a	NN	O	O
closely	NN	O	O
related	NN	O	O
protein	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
this	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
closest	NN	O	O
match	NN	O	O
to	NN	O	O
the	NN	O	O
known	NN	O	O
SP-1	NN	O	B-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
respective	NN	O	O
region	NN	O	O
is	NN	O	O
a	NN	O	O
TCCCCTCCCCT	NN	O	B-DNA
motif	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
this	NN	O	O
motif	NN	O	O
almost	NN	O	O
completely	NN	O	O
,	NN	O	O
and	NN	O	O
specifically	NN	O	O
,	NN	O	O
abolished	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
two	NN	O	O
low-mobility	NN	O	B-protein
complexes	NN	O	I-protein
and	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
95	NN	O	O
%	NN	O	O
decrease	NN	O	O
of	NN	O	O
constitutive	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	O
construct	NN	O	O
IL-1beta	NN	O	B-protein
(	NN	O	O
-170/+108	NN	O	O
)	NN	O	O
.	NN	O	O

Likewise	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
reporter	NN	O	O
construct	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
depended	NN	O	O
on	NN	O	O
the	NN	O	O
SP-1	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
so-far-unrecognized	NN	O	O
SP-1	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-1beta	NN	O	I-DNA
promoter	NN	O	I-DNA
may	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
in	NN	O	O
keratinocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
transformed	NN	O	O
phenotype	NN	O	O
and	NN	O	O
the	NN	O	O
type	NN	O	O
of	NN	O	O
viral	NN	O	O
latency	NN	O	O
in	NN	O	O
EBV	NN	O	B-cell_line
genome	NN	O	I-cell_line
positive	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
in	NN	O	O
B	NN	O	O
cell	NN	O	O
growth	NN	O	O
regulation	NN	O	O
by	NN	O	O
examining	NN	O	O
the	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
including	NN	O	O
both	NN	O	O
EBV	NN	O	B-DNA
genome	NN	O	I-DNA
negative	NN	O	O
and	NN	O	O
EBV	NN	O	B-cell_line
carrying	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Egr-1	NN	O	B-protein
expression	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
cellular	NN	O	O
phenotype	NN	O	O
and	NN	O	O
the	NN	O	O
specific	NN	O	O
pattern	NN	O	O
of	NN	O	O
viral	NN	O	O
latency	NN	O	O
established	NN	O	O
within	NN	O	O
the	NN	O	O
individual	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
gene	NN	O	O
is	NN	O	O
invariably	NN	O	O
associated	NN	O	O
with	NN	O	O
unrestricted	NN	O	O
expression	NN	O	O
of	NN	O	O
viral	NN	O	O
latent	NN	O	O
genes	NN	O	O
in	NN	O	O
all	NN	O	O
group	NN	O	B-cell_line
III	NN	O	I-cell_line
EBV	NN	O	I-cell_line
genome	NN	O	I-cell_line
carrying	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
Egr-1	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
abrogated	NN	O	O
in	NN	O	O
group	NN	O	B-cell_line
I	NN	O	I-cell_line
Burkitt	NN	O	I-cell_line
tumor	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
irrespective	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	B-DNA
genome	NN	O	I-DNA
carrying	NN	O	O
status	NN	O	O
.	NN	O	O

Activated	NN	O	O
viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
associated	NN	O	O
with	NN	O	O
phenotypic	NN	O	O
conversion	NN	O	O
of	NN	O	O
group	NN	O	B-cell_line
I	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
in	NN	O	O
to	NN	O	O
group	NN	O	B-cell_line
II	NN	O	I-cell_line
or	NN	O	O
III	NN	O	B-cell_line
restores	NN	O	O
the	NN	O	O
Egr-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Several	NN	O	O
forms	NN	O	O
of	NN	O	O
EGR-1	NN	O	B-protein
protein	NN	O	I-protein
are	NN	O	O
found	NN	O	O
within	NN	O	O
the	NN	O	O
different	NN	O	O
groups	NN	O	O
of	NN	O	O
cell	NN	O	O
lines	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
to	NN	O	O
DNA	NN	O	B-DNA
consensus	NN	O	I-DNA
sequences	NN	O	I-DNA
was	NN	O	O
investigated	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
time	NN	O	O
course	NN	O	O
analysis	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
the	NN	O	O
early	NN	O	O
steps	NN	O	O
of	NN	O	O
EBV	NN	O	O
infection	NN	O	O
in	NN	O	O
vitro	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
Egr-1	NN	O	B-protein
is	NN	O	O
upregulated	NN	O	O
within	NN	O	O
minutes	NN	O	O
from	NN	O	O
the	NN	O	O
initial	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
B	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
erythroid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
megakaryoblastic	NN	O	O
disorders	NN	O	O
.	NN	O	O

Currently	NN	O	O
available	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
erythroid	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
differentiation	NN	O	O
pathways	NN	O	O
are	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
each	NN	O	O
other	NN	O	O
,	NN	O	O
and	NN	O	O
there	NN	O	O
may	NN	O	O
exist	NN	O	O
progenitor	NN	O	B-cell_type
cells	NN	O	I-cell_type
common	NN	O	O
to	NN	O	O
those	NN	O	O
two	NN	O	O
lineages	NN	O	O
may	NN	O	O
exist	NN	O	O
.	NN	O	O

Acute	NN	O	O
megakaryoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
AML-M7	NN	O	O
)	NN	O	O
and	NN	O	O
transient	NN	O	O
myeloproliferative	NN	O	O
disorder	NN	O	O
in	NN	O	O
Down	NN	O	O
's	NN	O	O
syndrome	NN	O	O
(	NN	O	O
TMD	NN	O	O
)	NN	O	O
are	NN	O	O
characterized	NN	O	O
by	NN	O	O
rapid	NN	O	O
growth	NN	O	O
of	NN	O	O
abnormal	NN	O	B-cell_type
blast	NN	O	I-cell_type
cells	NN	O	I-cell_type
which	NN	O	O
express	NN	O	O
megakaryocytic	NN	O	B-protein
markers	NN	O	I-protein
.	NN	O	O

These	NN	O	O
blast	NN	O	B-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
lineage-specific	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
GATA-1	NN	O	B-protein
common	NN	O	O
to	NN	O	O
these	NN	O	O
lineages	NN	O	O
and	NN	O	O
frequently	NN	O	O
express	NN	O	O
erythroid-specific	NN	O	B-RNA
mRNAs	NN	O	I-RNA
such	NN	O	O
as	NN	O	O
gamma-globin	NN	O	B-protein
and	NN	O	O
erythroid	NN	O	B-protein
delta-aminolevulinate	NN	O	I-protein
synthase	NN	O	I-protein
(	NN	O	O
ALAS-E	NN	O	B-protein
)	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
blasts	NN	O	O
in	NN	O	O
M7	NN	O	O
and	NN	O	O
TMD	NN	O	O
cases	NN	O	O
have	NN	O	O
erythroid	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
blasts	NN	O	B-cell_type
in	NN	O	O
M7	NN	O	O
and	NN	O	O
TMD	NN	O	O
may	NN	O	O
correspond	NN	O	O
to	NN	O	O
progenitors	NN	O	B-cell_type
of	NN	O	O
both	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Stat3	NN	O	B-protein
recruitment	NN	O	O
by	NN	O	O
two	NN	O	O
distinct	NN	O	O
ligand-induced	NN	O	B-protein
,	NN	O	I-protein
tyrosine-phosphorylated	NN	O	I-protein
docking	NN	O	I-protein
sites	NN	O	I-protein
in	NN	O	O
the	NN	O	O
interleukin-10	NN	O	B-protein
receptor	NN	O	I-protein
intracellular	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Recent	NN	O	O
work	NN	O	O
has	NN	O	O
shown	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
induces	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
JAK	NN	O	B-protein
-STAT	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

To	NN	O	O
define	NN	O	O
the	NN	O	O
mechanism	NN	O	O
underlying	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
recruitment	NN	O	O
to	NN	O	O
the	NN	O	O
interleukin	NN	O	B-protein
10	NN	O	I-protein
(	NN	O	I-protein
IL-10	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
the	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
activated	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
in	NN	O	O
different	NN	O	O
cell	NN	O	O
populations	NN	O	O
were	NN	O	O
first	NN	O	O
defined	NN	O	O
using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
cells	NN	O	O
tested	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
activated	NN	O	O
Stat1	NN	O	B-protein
and	NN	O	O
Stat3	NN	O	B-protein
and	NN	O	O
induced	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
three	NN	O	O
distinct	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
that	NN	O	O
contained	NN	O	O
different	NN	O	O
combinations	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

IL-10	NN	O	B-protein
also	NN	O	O
activated	NN	O	O
Stat5	NN	O	B-protein
in	NN	O	O
Ba/F3	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
stably	NN	O	O
expressed	NN	O	O
the	NN	O	O
murine	NN	O	O
IL-10	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
a	NN	O	O
structure-function	NN	O	O
mutagenesis	NN	O	O
approach	NN	O	O
,	NN	O	O
two	NN	O	O
tyrosine	NN	O	O
residues	NN	O	O
(	NN	O	O
Tyr427	NN	O	O
and	NN	O	O
Tyr477	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
murine	NN	O	O
IL-10	NN	O	B-protein
receptor	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
redundantly	NN	O	O
required	NN	O	O
for	NN	O	O
receptor	NN	O	O
function	NN	O	O
and	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
but	NN	O	O
not	NN	O	O
for	NN	O	O
Stat1	NN	O	B-protein
or	NN	O	O
Stat5	NN	O	B-protein
.	NN	O	O

Twelve	NN	O	O
amino	NN	O	O
acid	NN	O	O
peptides	NN	O	O
encompassing	NN	O	O
either	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
tyrosine	NN	O	O
residues	NN	O	O
in	NN	O	O
phosphorylated	NN	O	B-protein
form	NN	O	I-protein
coprecipitated	NN	O	O
Stat3	NN	O	B-protein
but	NN	O	O
not	NN	O	O
Stat1	NN	O	B-protein
and	NN	O	O
blocked	NN	O	O
IL-10	NN	O	B-protein
-induced	NN	O	O
Stat3	NN	O	B-protein
phosphorylation	NN	O	O
in	NN	O	O
a	NN	O	O
cell-free	NN	O	O
system	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
tyrosine-phosphorylated	NN	O	O
peptides	NN	O	O
containing	NN	O	O
Tyr374	NN	O	O
or	NN	O	O
Tyr396	NN	O	O
did	NN	O	O
not	NN	O	O
interact	NN	O	O
with	NN	O	O
Stat3	NN	O	B-protein
or	NN	O	O
block	NN	O	O
Stat3	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Stat3	NN	O	B-protein
but	NN	O	O
not	NN	O	O
Stat1	NN	O	B-protein
or	NN	O	O
Stat5	NN	O	B-protein
is	NN	O	O
directly	NN	O	O
recruited	NN	O	O
to	NN	O	O
the	NN	O	O
ligand-activated	NN	O	B-protein
IL-10	NN	O	I-protein
receptor	NN	O	I-protein
by	NN	O	O
binding	NN	O	O
to	NN	O	O
specific	NN	O	O
but	NN	O	O
redundant	NN	O	O
receptor	NN	O	B-protein
intracellular	NN	O	I-protein
domain	NN	O	I-protein
sequences	NN	O	I-protein
containing	NN	O	O
phosphotyrosine	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
thus	NN	O	O
supports	NN	O	O
the	NN	O	O
concept	NN	O	O
that	NN	O	O
utilization	NN	O	O
of	NN	O	O
distinct	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
by	NN	O	O
different	NN	O	O
cytokine	NN	O	B-protein
receptors	NN	O	I-protein
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
particular	NN	O	O
ligand-activatable	NN	O	O
,	NN	O	O
tyrosine-containing	NN	O	B-protein
STAT	NN	O	I-protein
docking	NN	O	I-protein
sites	NN	O	I-protein
in	NN	O	O
receptor	NN	O	B-protein
intracellular	NN	O	I-protein
domains	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Silencing	NN	O	O
of	NN	O	O
human	NN	O	O
fetal	NN	O	O
globin	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
impaired	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
adult	NN	O	B-protein
beta-globin	NN	O	I-protein
gene	NN	O	I-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
EKLF	NN	O	B-protein
.	NN	O	O

Globin	NN	O	B-DNA
genes	NN	O	I-DNA
are	NN	O	O
subject	NN	O	O
to	NN	O	O
tissue-specific	NN	O	O
and	NN	O	O
developmental	NN	O	O
stage-specific	NN	O	O
regulation	NN	O	O
.	NN	O	O

A	NN	O	O
switch	NN	O	O
from	NN	O	O
human	NN	O	B-protein
fetal	NN	O	I-protein
(	NN	O	I-protein
gamma	NN	O	I-protein
)	NN	O	I-protein
-to	NN	O	O
adult	NN	O	B-protein
(	NN	O	I-protein
beta	NN	O	I-protein
)	NN	O	I-protein
-globin	NN	O	I-protein
expression	NN	O	O
occurs	NN	O	O
within	NN	O	O
erythroid	NN	O	B-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
adult	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

Previously	NN	O	O
we	NN	O	O
and	NN	O	O
others	NN	O	O
showed	NN	O	O
by	NN	O	O
targeted	NN	O	O
gene	NN	O	O
disruption	NN	O	O
that	NN	O	O
the	NN	O	O
zinc	NN	O	B-DNA
finger	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
erythroid	NN	O	B-protein
Kruppel-like	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EKLF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
mice	NN	O	O
,	NN	O	O
presumably	NN	O	O
through	NN	O	O
interaction	NN	O	O
with	NN	O	O
a	NN	O	O
high-affinity	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
EKLF	NN	O	B-protein
in	NN	O	O
the	NN	O	O
developmental	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
gamma-globin	NN	O	I-DNA
gene	NN	O	I-DNA
we	NN	O	O
interbred	NN	O	O
EKLF	NN	O	B-protein
heterozygotes	NN	O	I-protein
(	NN	O	O
+/-	NN	O	O
)	NN	O	O
with	NN	O	O
mice	NN	O	O
harboring	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
yeast	NN	O	I-DNA
artificial	NN	O	I-DNA
chromosome	NN	O	I-DNA
transgene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
EKLF	NN	O	B-protein
,	NN	O	O
while	NN	O	O
human	NN	O	B-protein
beta-globin	NN	O	I-protein
expression	NN	O	O
is	NN	O	O
dramatically	NN	O	O
reduced	NN	O	O
,	NN	O	O
gamma-globin	NN	O	B-RNA
transcripts	NN	O	I-RNA
are	NN	O	O
elevated	NN	O	O
approximately	NN	O	O
5-fold	NN	O	O
.	NN	O	O

Impaired	NN	O	O
silencing	NN	O	O
of	NN	O	O
gamma-globin	NN	O	B-protein
expression	NN	O	O
identifies	NN	O	O
EKLF	NN	O	B-protein
as	NN	O	O
the	NN	O	O
first	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
participating	NN	O	O
quantitatively	NN	O	O
in	NN	O	O
the	NN	O	O
gamma-globin	NN	O	B-protein
to	NN	O	O
beta-globin	NN	O	B-protein
switch	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
are	NN	O	O
compatible	NN	O	O
with	NN	O	O
a	NN	O	O
competitive	NN	O	O
model	NN	O	O
of	NN	O	O
switching	NN	O	O
in	NN	O	O
which	NN	O	O
EKLF	NN	O	B-protein
mediates	NN	O	O
an	NN	O	O
adult	NN	O	O
stage-specific	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
locus	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
that	NN	O	O
excludes	NN	O	O
the	NN	O	O
gamma-globin	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
zinc	NN	O	O
and	NN	O	O
related	NN	O	O
cations	NN	O	O
.	NN	O	O

Zinc	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
trace	NN	O	O
element	NN	O	O
for	NN	O	O
immune	NN	O	O
function	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
zinc	NN	O	O
addition	NN	O	O
in	NN	O	O
a	NN	O	O
serum-	NN	O	O
and	NN	O	O
lipopolysaccharide-free	NN	O	O
cell	NN	O	O
culture	NN	O	O
system	NN	O	O
leads	NN	O	O
to	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
levels	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
1	NN	O	I-protein
beta	NN	O	I-protein
(	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
)	NN	O	O
and	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
to	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
corresponding	NN	O	O
mRNA	NN	O	B-RNA
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

Structurally	NN	O	O
related	NN	O	O
divalent	NN	O	O
cations	NN	O	O
like	NN	O	O
cobalt	NN	O	O
,	NN	O	O
nickel	NN	O	O
,	NN	O	O
and	NN	O	O
mercury	NN	O	O
also	NN	O	O
partially	NN	O	O
increase	NN	O	O
monokine	NN	O	O
secretion	NN	O	O
but	NN	O	O
to	NN	O	O
a	NN	O	O
much	NN	O	O
lower	NN	O	O
and	NN	O	O
thus	NN	O	O
insignificant	NN	O	O
extent	NN	O	O
.	NN	O	O

They	NN	O	O
fail	NN	O	O
to	NN	O	O
induce	NN	O	O
mRNA	NN	O	B-RNA
of	NN	O	O
TNF-alpha	NN	O	B-protein
after	NN	O	O
3	NN	O	O
h	NN	O	O
of	NN	O	O
culture	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
monokine	NN	O	O
induction	NN	O	O
is	NN	O	O
a	NN	O	O
zinc-specific	NN	O	O
effect	NN	O	O
influenced	NN	O	O
by	NN	O	O
the	NN	O	O
physicochemical	NN	O	O
properties	NN	O	O
of	NN	O	O
the	NN	O	O
ion	NN	O	O
.	NN	O	O

Confirmation	NN	O	O
of	NN	O	O
the	NN	O	O
unique	NN	O	O
significance	NN	O	O
of	NN	O	O
zinc	NN	O	O
for	NN	O	O
immune	NN	O	O
function	NN	O	O
provides	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
specific	NN	O	O
zinc-mediated	NN	O	O
immune	NN	O	O
modulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

E3	NN	O	B-protein
,	NN	O	O
a	NN	O	O
hematopoietic-specific	NN	O	B-protein
transcript	NN	O	I-protein
directly	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
-induced	NN	O	O
maturation	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
myeloid	NN	O	O
development	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
differential	NN	O	O
hybridization	NN	O	O
analysis	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
library	NN	O	I-DNA
constructed	NN	O	O
from	NN	O	O
the	NN	O	O
murine	NN	O	O
RA-inducible	NN	O	B-cell_line
MPRO	NN	O	I-cell_line
promyelocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
to	NN	O	O
identify	NN	O	O
immediate-early	NN	O	B-DNA
genes	NN	O	I-DNA
induced	NN	O	O
by	NN	O	O
RA	NN	O	O
during	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

E3	NN	O	B-protein
,	NN	O	O
one	NN	O	O
of	NN	O	O
nine	NN	O	O
sequences	NN	O	O
identified	NN	O	O
,	NN	O	O
was	NN	O	O
upregulated	NN	O	O
in	NN	O	O
an	NN	O	O
immediate-early	NN	O	O
manner	NN	O	O
,	NN	O	O
with	NN	O	O
transcript	NN	O	O
levels	NN	O	O
peaking	NN	O	O
after	NN	O	O
60	NN	O	O
minutes	NN	O	O
exposure	NN	O	O
to	NN	O	O
RA	NN	O	O
.	NN	O	O

E3	NN	O	B-protein
transcripts	NN	O	I-protein
were	NN	O	O
RA-inducible	NN	O	O
in	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
an	NN	O	O
RA-resistant	NN	O	B-cell_line
subclone	NN	O	I-cell_line
,	NN	O	O
HL60R	NN	O	B-cell_line
,	NN	O	O
that	NN	O	O
harbors	NN	O	O
a	NN	O	O
mutated	NN	O	O
RAR	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
when	NN	O	O
HL60R	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
transduced	NN	O	O
with	NN	O	O
a	NN	O	O
functional	NN	O	O
copy	NN	O	O
of	NN	O	O
the	NN	O	O
RAR	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
RA	NN	O	O
induced	NN	O	O
a	NN	O	O
10-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
E3	NN	O	B-protein
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

E3	NN	O	B-protein
transcripts	NN	O	I-protein
are	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
myeloid	NN	O	B-cell_type
,	NN	O	I-cell_type
B-lymphoid	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
lineages	NN	O	I-cell_type
,	NN	O	O
absent	NN	O	O
in	NN	O	O
nonhematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
encode	NN	O	O
a	NN	O	O
highly	NN	O	O
hydrophobic	NN	O	O
,	NN	O	O
potentially	NN	O	O
phosphorylated	NN	O	O
polypeptide	NN	O	O
of	NN	O	O
unknown	NN	O	O
function	NN	O	O
with	NN	O	O
significant	NN	O	O
homology	NN	O	O
to	NN	O	O
a	NN	O	O
putative	NN	O	O
protein	NN	O	O
expressed	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
murine	NN	O	O
E3	NN	O	B-DNA
promoter	NN	O	I-DNA
harbors	NN	O	O
a	NN	O	O
single	NN	O	O
bipartite	NN	O	B-DNA
retinoic	NN	O	I-DNA
acid	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
which	NN	O	O
in	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
conferred	NN	O	O
RA	NN	O	O
sensitivity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
E3	NN	O	B-protein
is	NN	O	O
a	NN	O	O
hematopoietic-specific	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
an	NN	O	O
immediate	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
activated	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
during	NN	O	O
myelopoiesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Multifactor	NN	O	O
cis-dominant	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
anergized	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
underlying	NN	O	O
IL-2	NN	O	B-protein
transcriptional	NN	O	O
blockade	NN	O	O
in	NN	O	O
anergic	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
is	NN	O	O
not	NN	O	O
fully	NN	O	O
understood	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
whether	NN	O	O
an	NN	O	O
active	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
process	NN	O	O
occurs	NN	O	O
,	NN	O	O
we	NN	O	O
created	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
construct	NN	O	I-DNA
containing	NN	O	O
as	NN	O	O
an	NN	O	O
enhancer	NN	O	B-DNA
four	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-DNA
site	NN	O	I-DNA
and	NN	O	O
one	NN	O	O
copy	NN	O	O
of	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
(	NN	O	O
4X	NN	O	B-DNA
NF-AT-Oct	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

This	NN	O	O
construct	NN	O	O
was	NN	O	O
only	NN	O	O
slightly	NN	O	O
reduced	NN	O	O
(	NN	O	O
1.3-fold	NN	O	O
)	NN	O	O
in	NN	O	O
its	NN	O	O
expression	NN	O	O
when	NN	O	O
stimulated	NN	O	O
under	NN	O	O
anergic	NN	O	O
conditions	NN	O	O
,	NN	O	O
while	NN	O	O
a	NN	O	O
whole	NN	O	O
mouse	NN	O	O
IL-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
construct	NN	O	I-DNA
showed	NN	O	O
a	NN	O	O
reduction	NN	O	O
of	NN	O	O
4.3-fold	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
the	NN	O	O
-176	NN	O	B-DNA
to	NN	O	I-DNA
-96	NN	O	I-DNA
sequence	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
4X	NN	O	B-DNA
NF-AT-Oct	NN	O	I-DNA
construct	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
impart	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
be	NN	O	O
affected	NN	O	O
by	NN	O	O
anergy	NN	O	O
,	NN	O	O
but	NN	O	O
addition	NN	O	O
of	NN	O	O
the	NN	O	O
-236	NN	O	B-DNA
to	NN	O	I-DNA
-96	NN	O	I-DNA
sequence	NN	O	I-DNA
did	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
anergy	NN	O	O
is	NN	O	O
an	NN	O	O
active	NN	O	O
inhibitory	NN	O	O
process	NN	O	O
and	NN	O	O
that	NN	O	O
more	NN	O	O
than	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
-150	NN	O	B-DNA
AP-1	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
-152	NN	O	B-DNA
to	NN	O	I-DNA
-147	NN	O	I-DNA
)	NN	O	O
is	NN	O	O
required	NN	O	O
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
effect	NN	O	O
.	NN	O	O

Mutational	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
-236	NN	O	B-DNA
to	NN	O	I-DNA
-96	NN	O	I-DNA
sequence	NN	O	I-DNA
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
-130	NN	O	B-DNA
AP-1-like	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
-187	NN	O	B-DNA
to	NN	O	I-DNA
-181	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
the	NN	O	O
-150	NN	O	B-DNA
proximal	NN	O	I-DNA
AP-1	NN	O	I-DNA
site	NN	O	I-DNA
were	NN	O	O
necessary	NN	O	O
to	NN	O	O
observe	NN	O	O
anergy	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
-180	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
trans-activation	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
possible	NN	O	O
to	NN	O	O
confirm	NN	O	O
by	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
mouse	NN	O	B-DNA
IL-2	NN	O	I-DNA
enhancer	NN	O	I-DNA
that	NN	O	O
this	NN	O	O
site	NN	O	O
is	NN	O	O
absolutely	NN	O	O
essential	NN	O	O
for	NN	O	O
anergy	NN	O	O
induction	NN	O	O
.	NN	O	O

The	NN	O	O
simplest	NN	O	O
model	NN	O	O
to	NN	O	O
explain	NN	O	O
these	NN	O	O
results	NN	O	O
is	NN	O	O
that	NN	O	O
anergy	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
complex	NN	O	O
of	NN	O	O
multiple	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
exert	NN	O	O
a	NN	O	O
cis-acting	NN	O	O
dominant	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
trans-activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
due	NN	O	O
to	NN	O	O
defective	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
two	NN	O	O
male	NN	O	O
siblings	NN	O	O
.	NN	O	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
PBL	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
alloreactive	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
of	NN	O	O
two	NN	O	O
male	NN	O	O
infants	NN	O	O
born	NN	O	O
to	NN	O	O
consanguinous	NN	O	O
parents	NN	O	O
and	NN	O	O
presenting	NN	O	O
with	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
(	NN	O	O
SCID	NN	O	O
)	NN	O	O
showed	NN	O	O
a	NN	O	O
pronounced	NN	O	O
deficiency	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Although	NN	O	O
phenotypically	NN	O	O
normal	NN	O	O
,	NN	O	O
the	NN	O	O
proliferative	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
childrens	NN	O	O
'	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
strongly	NN	O	O
reduced	NN	O	O
but	NN	O	O
could	NN	O	O
be	NN	O	O
improved	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
both	NN	O	O
childrens	NN	O	O
'	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
unable	NN	O	O
to	NN	O	O
produce	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
IL-2	NN	O	B-protein
,	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

This	NN	O	O
multiple	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
deficiency	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
restored	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
or	NN	O	O
co-stimulatory	NN	O	O
signals	NN	O	O
provided	NN	O	O
by	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
APC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
could	NN	O	O
not	NN	O	O
be	NN	O	O
detected	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
activation-dependent	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
markers	NN	O	I-protein
CD25	NN	O	B-protein
and	NN	O	O
CD69	NN	O	B-protein
was	NN	O	O
within	NN	O	O
normal	NN	O	O
limits	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
functional	NN	O	O
defect	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
'	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
absence	NN	O	O
or	NN	O	O
abnormal	NN	O	O
binding	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
examine	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
Oct	NN	O	B-protein
,	NN	O	O
CREB	NN	O	B-protein
,	NN	O	O
SP1	NN	O	B-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
to	NN	O	O
their	NN	O	O
respective	NN	O	O
response	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Whereas	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
,	NN	O	O
Oct	NN	O	B-protein
,	NN	O	O
CREB	NN	O	B-protein
and	NN	O	O
SP1	NN	O	B-protein
displayed	NN	O	O
normal	NN	O	O
binding	NN	O	O
activities	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
to	NN	O	O
its	NN	O	O
IL-2	NN	O	B-protein
promoter	NN	O	O
response	NN	O	O
element	NN	O	O
was	NN	O	O
barely	NN	O	O
detectable	NN	O	O
both	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
T	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
this	NN	O	O
NF-AT	NN	O	B-protein
/DNA	NN	O	O
binding	NN	O	O
defect	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
multiple	NN	O	O
cytokine	NN	O	B-protein
deficiency	NN	O	O
and	NN	O	O
the	NN	O	O
SCID	NN	O	O
phenotype	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
infant	NN	O	O
brothers	NN	O	O
.	NN	O	O

-DOCSTART-	O

oriP	NN	O	B-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
EBNA	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus-immortalized	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

During	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
latent	NN	O	O
infection	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
six	NN	O	O
viral	NN	O	B-protein
nuclear	NN	O	I-protein
antigens	NN	O	I-protein
(	NN	O	O
EBNAs	NN	O	B-protein
)	NN	O	O
are	NN	O	O
expressed	NN	O	O
from	NN	O	O
one	NN	O	O
of	NN	O	O
two	NN	O	O
promoters	NN	O	B-DNA
,	NN	O	O
Cp	NN	O	B-DNA
or	NN	O	O
Wp	NN	O	B-DNA
,	NN	O	O
whose	NN	O	O
activities	NN	O	O
are	NN	O	O
mutually	NN	O	O
exclusive	NN	O	O
.	NN	O	O

Upon	NN	O	O
infection	NN	O	O
,	NN	O	O
Wp	NN	O	B-DNA
is	NN	O	O
initially	NN	O	O
active	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
a	NN	O	O
switch	NN	O	O
to	NN	O	O
Cp	NN	O	B-DNA
for	NN	O	O
the	NN	O	O
duration	NN	O	O
of	NN	O	O
latency	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
region	NN	O	O
upstream	NN	O	O
of	NN	O	O
Cp	NN	O	B-DNA
was	NN	O	O
analyzed	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cis	NN	O	O
elements	NN	O	O
involved	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
the	NN	O	O
EBNA	NN	O	B-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
in	NN	O	O
established	NN	O	O
in	NN	O	B-cell_line
vitro	NN	O	I-cell_line
immortalized	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
LCLs	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
determined	NN	O	O
that	NN	O	O
oriP	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
origin	NN	O	O
for	NN	O	O
episomal	NN	O	O
maintenance	NN	O	O
during	NN	O	O
latency	NN	O	O
,	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
efficient	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
from	NN	O	O
either	NN	O	O
Cp	NN	O	B-DNA
or	NN	O	O
Wp	NN	O	B-DNA
in	NN	O	O
LCLs	NN	O	B-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
some	NN	O	O
Burkitt	NN	O	B-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
the	NN	O	O
EBNA2-dependent	NN	O	B-DNA
enhancer	NN	O	I-DNA
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
Cp	NN	O	B-DNA
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
ca.	NN	O	O
two-	NN	O	O
to	NN	O	O
fivefold	NN	O	O
reduction	NN	O	O
in	NN	O	O
Cp	NN	O	B-DNA
activity	NN	O	O
in	NN	O	O
the	NN	O	O
LCLs	NN	O	B-cell_line
assayed	NN	O	O
.	NN	O	O

More	NN	O	O
extensive	NN	O	O
deletion	NN	O	O
of	NN	O	O
sequences	NN	O	O
upstream	NN	O	O
of	NN	O	O
Cp	NN	O	B-DNA
,	NN	O	O
including	NN	O	O
the	NN	O	O
EBNA2-dependent	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
nearly	NN	O	O
complete	NN	O	O
loss	NN	O	O
of	NN	O	O
Cp	NN	O	B-DNA
activity	NN	O	O
.	NN	O	O

This	NN	O	O
loss	NN	O	O
of	NN	O	O
activity	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
correlate	NN	O	O
with	NN	O	O
deletion	NN	O	O
of	NN	O	O
two	NN	O	O
CCAAT	NN	O	B-DNA
boxes	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
proximal	NN	O	B-DNA
CCAAT	NN	O	I-DNA
box	NN	O	I-DNA
located	NN	O	O
at	NN	O	O
bp	NN	O	B-DNA
-61	NN	O	I-DNA
to	NN	O	I-DNA
-65	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
distal	NN	O	O
CCAAT	NN	O	B-DNA
box	NN	O	I-DNA
located	NN	O	O
at	NN	O	O
bp	NN	O	O
-253	NN	O	B-DNA
to	NN	O	I-DNA
-257	NN	O	I-DNA
,	NN	O	O
upstream	NN	O	O
of	NN	O	O
Cp	NN	O	B-DNA
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
these	NN	O	O
cis	NN	O	B-DNA
elements	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
Cp	NN	O	B-DNA
activity	NN	O	O
is	NN	O	O
highly	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
properly	NN	O	O
positioned	NN	O	O
CCAAT	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
with	NN	O	O
the	NN	O	O
dependence	NN	O	O
on	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
CCAAT	NN	O	I-DNA
box	NN	O	I-DNA
apparent	NN	O	O
only	NN	O	O
when	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
CCAAT	NN	O	I-DNA
box	NN	O	I-DNA
was	NN	O	O
deleted	NN	O	O
or	NN	O	O
mutated	NN	O	O
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
response	NN	O	O
elements	NN	O	O
located	NN	O	O
at	NN	O	O
ca.	NN	O	B-DNA
bp	NN	O	I-DNA
-850	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
Cp	NN	O	B-DNA
did	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
significant	NN	O	O
loss	NN	O	O
in	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
general	NN	O	O
,	NN	O	O
deletions	NN	O	O
which	NN	O	O
diminished	NN	O	O
Cp	NN	O	B-DNA
activity	NN	O	O
resulted	NN	O	O
in	NN	O	O
induction	NN	O	O
of	NN	O	O
Wp	NN	O	B-DNA
activity	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
suppression	NN	O	O
of	NN	O	O
Wp	NN	O	B-DNA
activity	NN	O	O
by	NN	O	O
transcriptional	NN	O	O
interference	NN	O	O
from	NN	O	O
Cp	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
oriP	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
EBNA2-dependent	NN	O	B-DNA
enhancer	NN	O	I-DNA
as	NN	O	O
the	NN	O	O
major	NN	O	O
positive	NN	O	O
cis	NN	O	B-DNA
elements	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
regulating	NN	O	O
Cp	NN	O	B-DNA
activity	NN	O	O
in	NN	O	O
LCL	NN	O	B-cell_line
suggests	NN	O	O
that	NN	O	O
EBNA	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
is	NN	O	O
largely	NN	O	O
autoregulated	NN	O	O
by	NN	O	O
EBNA	NN	O	B-protein
1	NN	O	I-protein
and	NN	O	O
EBNA	NN	O	B-protein
2	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Protein-tyrosine	NN	O	O
kinase	NN	O	O
activation	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
lipopolysaccharide	NN	O	O
induction	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
1beta	NN	O	I-protein
and	NN	O	O
NFkappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
NFkappaB	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

In	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
interleukin	NN	O	B-protein
1beta	NN	O	I-protein
protein	NN	O	O
production	NN	O	O
and	NN	O	O
steady	NN	O	O
state	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
are	NN	O	O
increased	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
lipopolysaccharide	NN	O	O
,	NN	O	O
predominantly	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
increased	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-protein
1beta	NN	O	I-protein
gene	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
1beta	NN	O	I-protein
and	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
interleukin	NN	O	B-protein
6	NN	O	I-protein
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NFkappaB	NN	O	B-protein
.	NN	O	O

Since	NN	O	O
recent	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
lipopolysaccharide-induced	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activation	NN	O	O
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
NFkappaB	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
,	NN	O	O
we	NN	O	O
sought	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
NFkappaB	NN	O	B-protein
translocated	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
was	NN	O	O
active	NN	O	O
in	NN	O	O
stimulating	NN	O	O
transcription	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
pro-monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
,	NN	O	O
the	NN	O	O
lipopolysaccharide-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
1beta	NN	O	I-protein
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Tyrosine	NN	O	O
kinases	NN	O	O
are	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
lipopolysaccharide-mediated	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
to	NN	O	O
stimulate	NN	O	O
transcription	NN	O	O
is	NN	O	O
impaired	NN	O	O
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
of	NN	O	O
transcription	NN	O	O
is	NN	O	O
specific	NN	O	O
for	NN	O	O
NFkappaB	NN	O	B-protein
;	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
AP-1	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
is	NN	O	O
enhanced	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
,	NN	O	O
while	NN	O	O
lipopolysaccharide-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
requires	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
is	NN	O	O
not	NN	O	O
obligatory	NN	O	O
for	NN	O	O
lipopolysaccharide	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

-DOCSTART-	O

Chronic	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
infection	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
disrupts	NN	O	O
the	NN	O	O
autoregulatory	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
pathway	NN	O	O
via	NN	O	O
enhanced	NN	O	O
IkappaBalpha	NN	O	B-protein
degradation	NN	O	O
.	NN	O	O

Productive	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
infection	NN	O	O
causes	NN	O	O
sustained	NN	O	O
NF-kappaB	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
chronically	NN	O	O
infected	NN	O	O
monocytic	NN	O	O
cells	NN	O	O
.	NN	O	O

A	NN	O	O
direct	NN	O	O
temporal	NN	O	O
correlation	NN	O	O
exists	NN	O	O
between	NN	O	O
HIV	NN	O	O
infection	NN	O	O
and	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
myelomonoblastic	NN	O	B-cell_line
PLB-985	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
constitutive	NN	O	O
NF-kappaB	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
HIV1	NN	O	B-cell_type
-infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
turnover	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
protein	NN	O	O
,	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
double-stranded	NN	O	B-protein
RNA-dependent	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
PKR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
intracellular	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
subunits	NN	O	I-protein
in	NN	O	O
the	NN	O	O
PLB-985	NN	O	B-cell_line
and	NN	O	O
U937	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
cell	NN	O	I-cell_line
models	NN	O	I-cell_line
.	NN	O	O

HIV-1	NN	O	O
infection	NN	O	O
resulted	NN	O	O
in	NN	O	O
constitutive	NN	O	O
,	NN	O	O
low-level	NN	O	O
expression	NN	O	O
of	NN	O	O
type	NN	O	B-protein
1	NN	O	I-protein
interferon	NN	O	I-protein
(	NN	O	O
IFN	NN	O	B-protein
)	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
PKR	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
also	NN	O	O
detected	NN	O	O
in	NN	O	O
HIV-1-infected	NN	O	B-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
low-level	NN	O	O
IFN	NN	O	B-protein
production	NN	O	O
,	NN	O	O
since	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
anti-IFN-alpha/beta	NN	O	B-protein
antibody	NN	O	I-protein
to	NN	O	O
the	NN	O	O
cells	NN	O	O
decreased	NN	O	O
PKR	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
analysis	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
turnover	NN	O	O
demonstrated	NN	O	O
an	NN	O	O
increased	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
in	NN	O	O
HIV-1-infected	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
may	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
constitutive	NN	O	B-DNA
DNA	NN	O	I-DNA
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

A	NN	O	O
dramatic	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
intracellular	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
subunits	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	O
NF-kappaB2	NN	O	B-protein
p100	NN	O	I-protein
and	NN	O	O
a	NN	O	O
moderate	NN	O	O
increase	NN	O	O
in	NN	O	O
NF-kappaB2	NN	O	B-protein
p52	NN	O	I-protein
and	NN	O	O
RelA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
in	NN	O	O
HIV-1-infected	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
NF-kappaB1	NN	O	B-protein
p105/p50	NN	O	I-protein
levels	NN	O	O
were	NN	O	O
not	NN	O	O
altered	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
levels	NN	O	O
in	NN	O	O
uninfected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
induces	NN	O	O
IFN	NN	O	B-protein
production	NN	O	O
and	NN	O	O
PKR	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
contribute	NN	O	O
to	NN	O	O
enhanced	NN	O	O
IkappaBalpha	NN	O	B-protein
phosphorylation	NN	O	O
and	NN	O	O
subsequent	NN	O	O
degradation	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
subunits	NN	O	I-protein
may	NN	O	O
ultimately	NN	O	O
increase	NN	O	O
the	NN	O	O
intracellular	NN	O	O
pool	NN	O	O
of	NN	O	O
NF-kappaB/IkappaBalpha	NN	O	B-protein
by	NN	O	O
an	NN	O	O
autoregulatory	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Enhanced	NN	O	O
turnover	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
and	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
proteins	NN	O	I-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
chronically	NN	O	O
activated	NN	O	O
state	NN	O	O
of	NN	O	O
HIV-1-infected	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
c-Jun	NN	O	B-protein
delta-domain	NN	O	I-protein
inhibits	NN	O	O
neuroendocrine	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
a	NN	O	O
DNA	NN	O	O
sequence-	NN	O	O
and	NN	O	O
pituitary-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	O
and	NN	O	O
transformation	NN	O	O
activity	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
is	NN	O	O
governed	NN	O	O
by	NN	O	O
a	NN	O	O
27-amino	NN	O	B-protein
acid	NN	O	I-protein
regulatory	NN	O	I-protein
motif	NN	O	I-protein
,	NN	O	O
labeled	NN	O	O
the	NN	O	O
delta-domain	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
deleted	NN	O	O
in	NN	O	O
v-Jun	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
c-Jun	NN	O	B-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
the	NN	O	O
rat	NN	O	B-DNA
prolactin	NN	O	I-DNA
(	NN	O	I-DNA
rPRL	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
induced	NN	O	O
by	NN	O	O
either	NN	O	O
oncogenic	NN	O	B-protein
Ras	NN	O	I-protein
or	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
the	NN	O	O
structural	NN	O	O
and	NN	O	O
cell-specific	NN	O	O
requirements	NN	O	O
for	NN	O	O
this	NN	O	O
c-Jun	NN	O	B-protein
inhibitory	NN	O	O
response	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
c-Jun	NN	O	B-protein
inhibitory	NN	O	O
response	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
rPRL	NN	O	B-DNA
footprint	NN	O	I-DNA
II	NN	O	I-DNA
repressor	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
pituitary-specific	NN	O	O
and	NN	O	O
required	NN	O	O
the	NN	O	O
c-Jun	NN	O	B-protein
delta-domain	NN	O	I-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
alteration	NN	O	O
of	NN	O	O
any	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
features	NN	O	O
(	NN	O	O
e.g.	NN	O	O
,	NN	O	O
cis-element	NN	O	B-DNA
,	NN	O	O
trans-factor	NN	O	B-DNA
,	NN	O	O
or	NN	O	O
cell-specific	NN	O	O
background	NN	O	O
)	NN	O	O
switched	NN	O	O
c-Jun	NN	O	B-protein
to	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
of	NN	O	O
the	NN	O	O
rPRL	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
HeLa	NN	O	B-cell_line
nonpituitary	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
c-Jun	NN	O	B-protein
alone	NN	O	O
activated	NN	O	O
the	NN	O	O
rPRL	NN	O	B-DNA
promoter	NN	O	I-DNA
via	NN	O	O
the	NN	O	O
most	NN	O	O
proximal	NN	O	O
GHF-1/Pit-1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
footprint	NN	O	B-DNA
I	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
synergized	NN	O	O
with	NN	O	O
GHF-1	NN	O	B-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
recombinant	NN	O	O
GHF-1	NN	O	B-protein
interacted	NN	O	O
directly	NN	O	O
with	NN	O	O
c-Jun	NN	O	B-protein
but	NN	O	O
not	NN	O	O
c-Fos	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
important	NN	O	O
fundamental	NN	O	O
insights	NN	O	O
into	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
the	NN	O	O
c-Jun	NN	O	B-protein
delta-domain	NN	O	I-protein
functions	NN	O	O
as	NN	O	O
a	NN	O	O
modulatory	NN	O	O
switch	NN	O	O
and	NN	O	O
further	NN	O	O
imply	NN	O	O
that	NN	O	O
the	NN	O	O
functional	NN	O	O
role	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
is	NN	O	O
dictated	NN	O	O
by	NN	O	O
cell-specific	NN	O	O
influences	NN	O	O
and	NN	O	O
the	NN	O	O
delta-domain	NN	O	B-protein
motif	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Retinoic	NN	O	O
acid	NN	O	O
activates	NN	O	O
interferon	NN	O	B-protein
regulatory	NN	O	I-protein
factor-1	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

All-trans-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
is	NN	O	O
the	NN	O	O
drug	NN	O	O
of	NN	O	O
choice	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
.	NN	O	O

ATRA	NN	O	O
induces	NN	O	O
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
differentiation	NN	O	O
of	NN	O	O
APL	NN	O	B-cell_line
cells	NN	O	I-cell_line
into	NN	O	O
mature	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
ATRA-dependent	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
and	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
are	NN	O	O
not	NN	O	O
presently	NN	O	O
understood	NN	O	O
.	NN	O	O

The	NN	O	O
NB4	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
is	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
bone	NN	O	O
marrow	NN	O	O
of	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
APL	NN	O	O
during	NN	O	O
relapse	NN	O	O
,	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
model	NN	O	O
system	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
APL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Because	NN	O	O
interferon	NN	O	B-protein
(	NN	O	I-protein
IFN	NN	O	I-protein
)	NN	O	I-protein
regulatory	NN	O	I-protein
factors	NN	O	I-protein
(	NN	O	O
IRF-1	NN	O	B-protein
and	NN	O	O
IRF-2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
other	NN	O	O
IFN-inducible	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
regulate	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
ATRA	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
ATRA	NN	O	O
directly	NN	O	O
activates	NN	O	O
IRF-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
IRF-2	NN	O	B-protein
and	NN	O	O
2'-5	NN	O	B-protein
'	NN	O	I-protein
oligoadenylate	NN	O	I-protein
synthetase	NN	O	I-protein
(	NN	O	O
OAS	NN	O	O
)	NN	O	O
gene	NN	O	O
expression	NN	O	O
with	NN	O	O
slower	NN	O	O
kinetics	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
NB4	NN	O	O
cells	NN	O	O
,	NN	O	O
ATRA	NN	O	O
also	NN	O	O
activated	NN	O	O
IRF-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
HL-60	NN	O	B-cell_line
,	NN	O	O
U937	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
all	NN	O	O
respond	NN	O	O
to	NN	O	O
ATRA	NN	O	O
by	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
.	NN	O	O

A	NN	O	O
more	NN	O	O
than	NN	O	O
additive	NN	O	O
increase	NN	O	O
in	NN	O	O
IRF-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
seen	NN	O	O
with	NN	O	O
ATRA	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

ATRA	NN	O	O
did	NN	O	O
not	NN	O	O
activate	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
kappa	NN	O	O
B	NN	O	O
or	NN	O	O
signal	NN	O	O
transducer	NN	O	O
and	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	O
(	NN	O	O
STAT	NN	O	O
)	NN	O	O
activation	NN	O	O
pathways	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
an	NN	O	O
alternate	NN	O	O
mechanism	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
IRF-1	NN	O	B-DNA
gene	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

The	NN	O	O
ATRA-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
IRF-1	NN	O	B-protein
,	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	O
and	NN	O	O
repressor	NN	O	O
of	NN	O	O
transformation	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
ATRA-induced	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
synergistic	NN	O	O
actions	NN	O	O
of	NN	O	O
ATRA	NN	O	O
and	NN	O	O
IFNs	NN	O	B-protein
in	NN	O	O
myeloid	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Recombinant	NN	O	O
NFAT1	NN	O	B-protein
(	NN	O	O
NFATp	NN	O	O
)	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
calcineurin	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
mediates	NN	O	O
transcription	NN	O	O
of	NN	O	O
several	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
other	NN	O	O
genes	NN	O	O
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
two	NN	O	O
new	NN	O	O
isoforms	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NFAT1	NN	O	B-protein
(	NN	O	O
previously	NN	O	O
termed	NN	O	O
NFATp	NN	O	B-protein
)	NN	O	O
that	NN	O	O
are	NN	O	O
the	NN	O	O
predominant	NN	O	O
isoforms	NN	O	O
expressed	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
and	NN	O	I-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

When	NN	O	O
expressed	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
recombinant	NN	O	O
NFAT1	NN	O	B-protein
is	NN	O	O
regulated	NN	O	O
,	NN	O	O
as	NN	O	O
expected	NN	O	O
,	NN	O	O
by	NN	O	O
the	NN	O	O
calmodulin-dependent	NN	O	B-protein
phosphatase	NN	O	I-protein
calcineurin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
its	NN	O	O
function	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
agent	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
.	NN	O	O

Transactivation	NN	O	O
by	NN	O	O
recombinant	NN	O	O
NFAT1	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
requires	NN	O	O
dual	NN	O	O
stimulation	NN	O	O
with	NN	O	O
ionomycin	NN	O	O
and	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
;	NN	O	O
this	NN	O	O
activity	NN	O	O
is	NN	O	O
potentiated	NN	O	O
by	NN	O	O
coexpression	NN	O	O
of	NN	O	O
constitutively	NN	O	O
active	NN	O	O
calcineurin	NN	O	B-protein
and	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
CsA	NN	O	O
.	NN	O	O

Immunocytochemical	NN	O	O
analysis	NN	O	O
indicates	NN	O	O
that	NN	O	O
recombinant	NN	O	O
NFAT1	NN	O	B-protein
localizes	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
transiently	NN	O	B-cell_type
transfected	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
translocates	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
in	NN	O	O
a	NN	O	O
CsA-sensitive	NN	O	O
manner	NN	O	O
following	NN	O	O
ionomycin	NN	O	O
stimulation	NN	O	O
.	NN	O	O

When	NN	O	O
expressed	NN	O	O
in	NN	O	O
COS	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
however	NN	O	O
,	NN	O	O
NFAT1	NN	O	B-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
transactivation	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
regulated	NN	O	O
correctly	NN	O	O
:	NN	O	O
its	NN	O	O
subcellular	NN	O	O
localization	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
function	NN	O	O
are	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
COS	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
ionomycin	NN	O	O
and	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
.	NN	O	O

Recombinant	NN	O	O
NFAT1	NN	O	B-protein
can	NN	O	O
mediate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
,	NN	O	O
interleukin-4	NN	O	O
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
and	NN	O	O
granulocyte-macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
promoters	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
NFAT1	NN	O	B-protein
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
CsA-sensitive	NN	O	O
transcription	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	B-DNA
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-protein
TAFII	NN	O	I-protein
105	NN	O	I-protein
is	NN	O	O
a	NN	O	O
cell	NN	O	B-protein
type-specific	NN	O	I-protein
TFIID	NN	O	I-protein
subunit	NN	O	I-protein
related	NN	O	O
to	NN	O	O
hTAFII130	NN	O	B-protein
.	NN	O	O

We	NN	O	O
previously	NN	O	O
characterized	NN	O	O
Drosophila	NN	O	B-protein
and	NN	O	I-protein
human	NN	O	I-protein
TAF	NN	O	I-protein
subunits	NN	O	I-protein
that	NN	O	O
make	NN	O	O
up	NN	O	O
the	NN	O	O
core	NN	O	O
TFIID	NN	O	B-protein
complex	NN	O	I-protein
found	NN	O	O
in	NN	O	O
all	NN	O	O
cells	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
differentiated	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
contain	NN	O	O
a	NN	O	O
novel	NN	O	O
substoichiometric	NN	O	O
TAF	NN	O	B-protein
of	NN	O	O
105	NN	O	O
kDa	NN	O	O
not	NN	O	O
found	NN	O	O
associated	NN	O	O
with	NN	O	O
TFIID	NN	O	B-protein
isolated	NN	O	O
from	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

The	NN	O	O
cDNA	NN	O	B-DNA
encoding	NN	O	O
hTAFII105	NN	O	B-protein
reveals	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
C-terminal	NN	O	B-protein
domain	NN	O	I-protein
shared	NN	O	O
by	NN	O	O
hTAFII130	NN	O	B-protein
and	NN	O	O
oTAFII110	NN	O	B-protein
,	NN	O	O
while	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
coactivator	NN	O	I-protein
domain	NN	O	I-protein
has	NN	O	O
diverged	NN	O	O
significantly	NN	O	O
.	NN	O	O

All	NN	O	O
cells	NN	O	O
tested	NN	O	O
express	NN	O	O
TAFII105	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
but	NN	O	O
only	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
contain	NN	O	O
significant	NN	O	O
levels	NN	O	O
of	NN	O	O
protein	NN	O	O
associated	NN	O	O
with	NN	O	O
TFIID	NN	O	B-protein
.	NN	O	O

Transient	NN	O	O
overexpression	NN	O	O
of	NN	O	O
hTAFII105	NN	O	B-protein
selectively	NN	O	O
squelches	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
some	NN	O	O
genes	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
properties	NN	O	O
suggest	NN	O	O
that	NN	O	O
TAFII105	NN	O	B-protein
is	NN	O	O
a	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
subunit	NN	O	O
of	NN	O	O
TFIID	NN	O	B-protein
that	NN	O	O
may	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
mediating	NN	O	O
transcription	NN	O	O
by	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
activators	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Interstitial	NN	O	O
deletion	NN	O	O
constitutes	NN	O	O
the	NN	O	O
major	NN	O	O
mechanism	NN	O	O
for	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
20q	NN	O	I-DNA
in	NN	O	O
polycythemia	NN	O	O
vera	NN	O	O
.	NN	O	O

An	NN	O	O
acquired	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	O
arm	NN	O	O
of	NN	O	O
chromosome	NN	O	B-DNA
20	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
recurrent	NN	O	O
abnormality	NN	O	O
in	NN	O	O
myeloproliferative	NN	O	O
disorders	NN	O	O
,	NN	O	O
particularly	NN	O	O
polycythemia	NN	O	O
vera	NN	O	O
and	NN	O	O
myelodysplastic	NN	O	O
syndromes	NN	O	O
.	NN	O	O

The	NN	O	O
association	NN	O	O
of	NN	O	O
20q	NN	O	B-DNA
deletions	NN	O	I-DNA
with	NN	O	O
myeloid	NN	O	O
``	NN	O	O
stem	NN	O	O
cell	NN	O	O
''	NN	O	O
disorders	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
deletions	NN	O	O
mark	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
genes	NN	O	O
,	NN	O	O
loss	NN	O	O
or	NN	O	O
inactivation	NN	O	O
of	NN	O	O
which	NN	O	O
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
performed	NN	O	O
a	NN	O	O
detailed	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
20q	NN	O	B-DNA
deletions	NN	O	I-DNA
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
(	NN	O	I-cell_type
PB	NN	O	I-cell_type
)	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
and	NN	O	O
defined	NN	O	O
a	NN	O	O
commonly	NN	O	B-DNA
deleted	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
16	NN	O	O
to	NN	O	O
21	NN	O	O
centimorgan	NN	O	O
(	NN	O	O
cM	NN	O	O
)	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
reduce	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
the	NN	O	O
common	NN	O	O
deleted	NN	O	O
region	NN	O	O
we	NN	O	O
have	NN	O	O
searched	NN	O	O
for	NN	O	O
small	NN	O	O
deletions	NN	O	O
or	NN	O	O
mitotic	NN	O	O
recombination	NN	O	O
events	NN	O	O
,	NN	O	O
neither	NN	O	O
of	NN	O	O
which	NN	O	O
would	NN	O	O
be	NN	O	O
detected	NN	O	O
by	NN	O	O
conventional	NN	O	O
cytogenetics	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
48	NN	O	O
patients	NN	O	O
with	NN	O	O
polycythemia	NN	O	O
vera	NN	O	O
and	NN	O	O
four	NN	O	O
patients	NN	O	O
with	NN	O	O
idiopathic	NN	O	O
myelofibrosis	NN	O	O
.	NN	O	O

In	NN	O	O
each	NN	O	O
case	NN	O	O
,	NN	O	O
cytogenetic	NN	O	O
analysis	NN	O	O
had	NN	O	O
either	NN	O	O
failed	NN	O	O
or	NN	O	O
had	NN	O	O
shown	NN	O	O
no	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
chromosome	NN	O	B-DNA
20	NN	O	I-DNA
.	NN	O	O

Seventeen	NN	O	O
microsatellite	NN	O	B-DNA
markers	NN	O	I-DNA
that	NN	O	O
span	NN	O	O
the	NN	O	O
common	NN	O	O
deleted	NN	O	O
region	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
search	NN	O	O
for	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
in	NN	O	O
granulocyte	NN	O	B-DNA
DNA	NN	O	I-DNA
.	NN	O	O

No	NN	O	O
instance	NN	O	O
of	NN	O	O
microsatellite	NN	O	O
instability	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
880	NN	O	O
comparisons	NN	O	O
of	NN	O	O
granulocyte	NN	O	B-DNA
and	NN	O	I-DNA
T-cell	NN	O	I-DNA
DNA	NN	O	I-DNA
.	NN	O	O

Granulocyte	NN	O	O
DNA	NN	O	O
from	NN	O	O
four	NN	O	O
patients	NN	O	O
exhibited	NN	O	O
allele	NN	O	O
loss	NN	O	O
on	NN	O	O
20q	NN	O	B-DNA
.	NN	O	O

In	NN	O	O
each	NN	O	O
case	NN	O	O
the	NN	O	O
allele	NN	O	O
loss	NN	O	O
was	NN	O	O
caused	NN	O	O
by	NN	O	O
an	NN	O	O
interstitial	NN	O	O
deletion	NN	O	O
because	NN	O	O
heterozygosity	NN	O	O
at	NN	O	O
distal	NN	O	B-DNA
markers	NN	O	I-DNA
was	NN	O	O
retained	NN	O	O
and	NN	O	O
because	NN	O	O
quantitative	NN	O	O
Southern	NN	O	O
blotting	NN	O	O
demonstrated	NN	O	O
hemizygosity	NN	O	O
.	NN	O	O

Loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
in	NN	O	O
PB	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
would	NN	O	O
be	NN	O	O
masked	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
significant	NN	O	O
numbers	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
not	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
malignant	NN	O	B-cell_line
clone	NN	O	I-cell_line
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
human	NN	O	B-protein
androgen	NN	O	I-protein
receptor	NN	O	I-protein
assay	NN	O	O
(	NN	O	O
HUMARA	NN	O	O
)	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
granulocyte	NN	O	O
clonality	NN	O	O
.	NN	O	O

In	NN	O	O
21	NN	O	O
of	NN	O	O
27	NN	O	O
informative	NN	O	O
female	NN	O	O
patients	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
granulocytes	NN	O	B-cell_type
were	NN	O	O
clonally	NN	O	O
derived	NN	O	O
.	NN	O	O

In	NN	O	O
5	NN	O	O
patients	NN	O	O
the	NN	O	O
granulocytes	NN	O	B-cell_type
appeared	NN	O	O
polyclonal	NN	O	O
and	NN	O	O
in	NN	O	O
1	NN	O	O
patient	NN	O	O
unilateral	NN	O	O
X	NN	O	O
inactivation	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
both	NN	O	O
granulocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
vast	NN	O	O
majority	NN	O	O
of	NN	O	O
patients	NN	O	O
presented	NN	O	O
here	NN	O	O
,	NN	O	O
the	NN	O	O
failure	NN	O	O
to	NN	O	O
detect	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
attributed	NN	O	O
to	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_line
polyclonal	NN	O	I-cell_line
granulocytes	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
results	NN	O	O
therefore	NN	O	O
show	NN	O	O
that	NN	O	O
allele	NN	O	O
loss	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
20q	NN	O	I-DNA
in	NN	O	O
polycythemia	NN	O	O
vera	NN	O	O
does	NN	O	O
not	NN	O	O
commonly	NN	O	O
involve	NN	O	O
mitotic	NN	O	O
recombination	NN	O	O
or	NN	O	O
chromosome	NN	O	B-DNA
loss	NN	O	O
and	NN	O	O
that	NN	O	O
microsatellite	NN	O	O
instability	NN	O	O
is	NN	O	O
a	NN	O	O
rare	NN	O	O
event	NN	O	O
in	NN	O	O
this	NN	O	O
disorder	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin-6	NN	O	B-protein
promotes	NN	O	O
multiple	NN	O	O
myeloma	NN	O	O
cell	NN	O	O
growth	NN	O	O
via	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
retinoblastoma	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Interleukin-6	NN	O	B-protein
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
mediates	NN	O	O
autocrine	NN	O	O
and	NN	O	O
paracrine	NN	O	O
growth	NN	O	O
of	NN	O	O
multiple	NN	O	B-cell_line
myeloma	NN	O	I-cell_line
(	NN	O	I-cell_line
MM	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
inhibits	NN	O	O
tumor	NN	O	O
cell	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Abnormalities	NN	O	O
of	NN	O	O
retinoblastoma	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
pRB	NN	O	B-protein
)	NN	O	O
and	NN	O	O
mutations	NN	O	O
of	NN	O	O
RB	NN	O	O
gene	NN	O	O
have	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
up	NN	O	O
to	NN	O	O
70	NN	O	O
%	NN	O	O
of	NN	O	O
MM	NN	O	O
patients	NN	O	O
and	NN	O	O
80	NN	O	O
%	NN	O	O
of	NN	O	O
MM-derived	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Because	NN	O	O
dephosphorylated	NN	O	B-protein
(	NN	O	I-protein
activated	NN	O	I-protein
)	NN	O	I-protein
pRB	NN	O	I-protein
blocks	NN	O	O
transition	NN	O	O
from	NN	O	O
G1	NN	O	O
to	NN	O	O
S	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
whereas	NN	O	O
phosphorylated	NN	O	B-protein
(	NN	O	I-protein
inactivated	NN	O	I-protein
)	NN	O	I-protein
pRB	NN	O	I-protein
releases	NN	O	O
this	NN	O	O
growth	NN	O	O
arrest	NN	O	O
,	NN	O	O
we	NN	O	O
characterized	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
pRB	NN	O	B-protein
in	NN	O	O
IL-6	NN	O	B-protein
-mediated	NN	O	O
MM	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

Both	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
dephosphorylated	NN	O	O
pRB	NN	O	B-protein
were	NN	O	O
expressed	NN	O	O
in	NN	O	O
all	NN	O	O
serum-starved	NN	O	O
MM	NN	O	O
patient	NN	O	O
cells	NN	O	O
and	NN	O	O
MM-derived	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
pRB	NN	O	B-protein
was	NN	O	O
predominantly	NN	O	O
in	NN	O	O
its	NN	O	O
phosphorylated	NN	O	O
form	NN	O	O
.	NN	O	O

In	NN	O	O
MM	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
proliferated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
exogenous	NN	O	O
IL-6	NN	O	B-protein
downregulated	NN	O	O
dephosphorylated	NN	O	B-protein
pRB	NN	O	I-protein
and	NN	O	O
decreased	NN	O	O
dephosphorylated	NN	O	O
pRB-E2F	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
culture	NN	O	O
of	NN	O	O
MM	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
RB	NN	O	O
antisense	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
RB	NN	O	O
sense	NN	O	O
,	NN	O	O
oligonucleotide	NN	O	O
(	NN	O	O
ODN	NN	O	O
)	NN	O	O
triggered	NN	O	O
IL-6	NN	O	B-protein
secretion	NN	O	O
and	NN	O	O
proliferation	NN	O	O
in	NN	O	O
MM	NN	O	B-cell_line
cells	NN	O	I-cell_line
;	NN	O	O
however	NN	O	O
,	NN	O	O
proliferation	NN	O	O
was	NN	O	O
only	NN	O	O
partially	NN	O	O
inhibited	NN	O	O
by	NN	O	O
neutralizing	NN	O	O
anti-IL-6	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
MoAb	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
MM	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
normal	NN	O	O
splenic	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
dephosphorylated	NN	O	B-protein
pRB	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
CD40	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
CD40L	NN	O	B-protein
)	NN	O	O
triggers	NN	O	O
a	NN	O	O
shift	NN	O	O
from	NN	O	O
dephosphorylated	NN	O	O
to	NN	O	O
phosphorylated	NN	O	O
pRB	NN	O	B-protein
and	NN	O	O
proliferation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
exogenous	NN	O	O
IL-6	NN	O	B-protein
to	NN	O	O
CD40L	NN	O	B-protein
-treated	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
does	NN	O	O
not	NN	O	O
alter	NN	O	O
either	NN	O	O
pRB	NN	O	B-protein
or	NN	O	O
proliferation	NN	O	O
,	NN	O	O
as	NN	O	O
observed	NN	O	O
in	NN	O	O
MM	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
phosphorylated	NN	O	B-protein
pRB	NN	O	I-protein
is	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
MM	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
that	NN	O	O
IL-6	NN	O	B-protein
further	NN	O	O
shifts	NN	O	O
pRB	NN	O	B-protein
from	NN	O	O
its	NN	O	O
dephosphorylated	NN	O	O
to	NN	O	O
its	NN	O	O
phosphorylated	NN	O	O
form	NN	O	O
,	NN	O	O
thereby	NN	O	O
promoting	NN	O	O
MM	NN	O	O
cell	NN	O	O
growth	NN	O	O
via	NN	O	O
two	NN	O	O
mechanisms	NN	O	O
;	NN	O	O
by	NN	O	O
decreasing	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
E2F	NN	O	O
bound	NN	O	O
by	NN	O	O
dephosphorylated	NN	O	B-protein
pRB	NN	O	I-protein
due	NN	O	O
to	NN	O	O
reduced	NN	O	O
dephosphorylated	NN	O	B-protein
pRB	NN	O	I-protein
,	NN	O	O
thereby	NN	O	O
releasing	NN	O	O
growth	NN	O	O
arrest	NN	O	O
;	NN	O	O
and	NN	O	O
by	NN	O	O
upregulating	NN	O	O
IL-6	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
MM	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
related	NN	O	O
IL-6	NN	O	B-protein
-mediated	NN	O	O
autocrine	NN	O	O
tumor	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

-DOCSTART-	O

Soluble	NN	O	B-protein
factors	NN	O	I-protein
secreted	NN	O	O
by	NN	O	O
activated	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
modulate	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	B-protein
cytokine	NN	O	I-protein
TGF-beta	NN	O	B-protein
2	NN	O	I-protein
in	NN	O	O
glial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Coordination	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
to	NN	O	O
injury	NN	O	O
or	NN	O	O
disease	NN	O	O
in	NN	O	O
the	NN	O	O
brain	NN	O	O
is	NN	O	O
postulated	NN	O	O
to	NN	O	O
involve	NN	O	O
bi-directional	NN	O	O
discourse	NN	O	O
between	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
and	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
.	NN	O	O

This	NN	O	O
cross	NN	O	O
communication	NN	O	O
involves	NN	O	O
soluble	NN	O	O
mediators	NN	O	O
,	NN	O	O
including	NN	O	O
various	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
and	NN	O	O
neuropeptides	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
supernatant	NN	O	O
from	NN	O	O
activated	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
is	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
potent	NN	O	O
cytokine	NN	O	B-protein
,	NN	O	O
TGF-beta	NN	O	B-protein
2	NN	O	I-protein
in	NN	O	O
glial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
activating	NN	O	O
stimulus	NN	O	O
invokes	NN	O	O
signaling	NN	O	O
mechanisms	NN	O	O
distinct	NN	O	O
from	NN	O	O
known	NN	O	O
kinase	NN	O	O
or	NN	O	O
protease	NN	O	O
pathways	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
TGF-beta	NN	O	B-protein
2	NN	O	I-protein
transcription	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
binding	NN	O	O
activity	NN	O	O
for	NN	O	O
an	NN	O	O
80	NN	O	B-protein
kDA	NN	O	I-protein
glial	NN	O	I-protein
labile	NN	O	I-protein
repressor	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
GLRP	NN	O	B-protein
,	NN	O	O
to	NN	O	O
a	NN	O	O
responsive	NN	O	O
region	NN	O	O
within	NN	O	O
the	NN	O	O
TFG-beta	NN	O	B-DNA
2	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Although	NN	O	O
GLRP	NN	O	B-protein
shares	NN	O	O
some	NN	O	O
characteristics	NN	O	O
with	NN	O	O
the	NN	O	O
inducible	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
,	NN	O	O
it	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
distinct	NN	O	O
from	NN	O	O
known	NN	O	O
AP-1	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
along	NN	O	O
with	NN	O	O
previous	NN	O	O
observations	NN	O	O
demonstrating	NN	O	O
the	NN	O	O
potent	NN	O	O
immunosuppressive	NN	O	O
activity	NN	O	O
of	NN	O	O
TGF-beta	NN	O	B-protein
2	NN	O	I-protein
,	NN	O	O
support	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
a	NN	O	O
feedback	NN	O	O
mechanism	NN	O	O
between	NN	O	O
the	NN	O	O
activated	NN	O	O
T-lymphocytes	NN	O	B-cell_type
and	NN	O	O
astrocytes	NN	O	B-cell_type
via	NN	O	O
TGF-beta	NN	O	B-protein
2	NN	O	I-protein
to	NN	O	O
regulate	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Various	NN	O	O
modes	NN	O	O
of	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
protein	NN	O	I-protein
-mediated	NN	O	O
regulation	NN	O	O
of	NN	O	O
murine	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
transcription	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
transcriptionally	NN	O	B-DNA
regulatory	NN	O	I-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
lymphomagenic	NN	O	O
Akv	NN	O	O
and	NN	O	O
SL3-3	NN	O	O
murine	NN	O	O
leukemia	NN	O	O
retroviruses	NN	O	O
(	NN	O	O
MLVs	NN	O	O
)	NN	O	O
contain	NN	O	O
two	NN	O	O
types	NN	O	O
of	NN	O	O
E-box	NN	O	B-DNA
consensus	NN	O	I-DNA
motifs	NN	O	I-DNA
,	NN	O	O
CAGATG	NN	O	O
.	NN	O	O

One	NN	O	O
type	NN	O	O
,	NN	O	O
EA/S	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
other	NN	O	O
,	NN	O	O
E	NN	O	B-DNA
(	NN	O	I-DNA
gre	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
located	NN	O	O
in	NN	O	O
a	NN	O	O
tandem	NN	O	B-DNA
repeat	NN	O	I-DNA
with	NN	O	O
enhancer	NN	O	O
properties	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
requirements	NN	O	O
of	NN	O	O
the	NN	O	O
individual	NN	O	O
E-boxes	NN	O	B-DNA
in	NN	O	O
MLV	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
.	NN	O	O

In	NN	O	O
lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
only	NN	O	O
,	NN	O	O
the	NN	O	O
E	NN	O	B-protein
(	NN	O	I-protein
gre	NN	O	I-protein
)	NN	O	I-protein
-binding	NN	O	I-protein
protein	NN	O	I-protein
complexes	NN	O	I-protein
included	NN	O	O
ALF1	NN	O	B-protein
or	NN	O	O
HEB	NN	O	B-protein
and	NN	O	O
E2A	NN	O	B-protein
basic	NN	O	I-protein
helix-loop-helix	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Ectopic	NN	O	O
ALF1	NN	O	B-protein
and	NN	O	O
E2A	NN	O	B-protein
proteins	NN	O	I-protein
required	NN	O	O
intact	NN	O	O
E	NN	O	B-DNA
(	NN	O	I-DNA
gre	NN	O	I-DNA
)	NN	O	I-DNA
motifs	NN	O	I-DNA
for	NN	O	O
mediating	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

ALF1	NN	O	B-protein
transactivated	NN	O	O
transcription	NN	O	O
of	NN	O	O
Akv	NN	O	O
MLV	NN	O	O
through	NN	O	O
the	NN	O	O
two	NN	O	O
E	NN	O	B-DNA
(	NN	O	I-DNA
gre	NN	O	I-DNA
)	NN	O	I-DNA
motifs	NN	O	I-DNA
equally	NN	O	O
,	NN	O	O
whereas	NN	O	O
E2A	NN	O	B-protein
protein	NN	O	I-protein
required	NN	O	O
the	NN	O	O
promoter-proximal	NN	O	O
E	NN	O	B-DNA
(	NN	O	I-DNA
gre	NN	O	I-DNA
)	NN	O	I-DNA
motif	NN	O	O
.	NN	O	O

In	NN	O	O
T-	NN	O	B-cell_line
and	NN	O	I-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
E	NN	O	B-DNA
(	NN	O	I-DNA
gre	NN	O	I-DNA
)	NN	O	I-DNA
motifs	NN	O	I-DNA
were	NN	O	O
of	NN	O	O
major	NN	O	O
importance	NN	O	O
for	NN	O	O
Akv	NN	O	O
MLV	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
EA/S	NN	O	B-DNA
motif	NN	O	I-DNA
had	NN	O	O
some	NN	O	O
effect	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
neither	NN	O	O
E	NN	O	B-DNA
(	NN	O	I-DNA
gre	NN	O	I-DNA
)	NN	O	I-DNA
nor	NN	O	O
EA/S	NN	O	B-DNA
motifs	NN	O	I-DNA
contributed	NN	O	O
pronouncedly	NN	O	O
to	NN	O	O
Akv	NN	O	O
MLV	NN	O	O
transcription	NN	O	O
in	NN	O	O
NIH	NN	O	B-cell_line
3T3	NN	O	I-cell_line
cells	NN	O	I-cell_line
lacking	NN	O	O
DNA-binding	NN	O	O
ALF1	NN	O	B-protein
or	NN	O	O
HEB	NN	O	B-protein
and	NN	O	O
E2A	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Id1	NN	O	O
protein	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
repress	NN	O	O
ALF1	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
ectopic	NN	O	B-protein
Id1	NN	O	I-protein
repressed	NN	O	O
E	NN	O	B-DNA
(	NN	O	I-DNA
gre	NN	O	I-DNA
)	NN	O	I-DNA
-directed	NN	O	O
but	NN	O	O
not	NN	O	O
EA/S	NN	O	B-DNA
-directed	NN	O	O
MLV	NN	O	O
transcription	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
E	NN	O	B-DNA
(	NN	O	I-DNA
gre	NN	O	I-DNA
)	NN	O	I-DNA
motifs	NN	O	O
and	NN	O	O
interacting	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
important	NN	O	O
determinants	NN	O	O
for	NN	O	O
MLV	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
lymphocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Reversal	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
the	NN	O	O
leukaemia-associated	NN	O	O
E2A-HLF	NN	O	B-protein
chimaeric	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
E2A-HLF	NN	O	B-DNA
(	NN	O	I-DNA
for	NN	O	I-DNA
hepatic	NN	O	I-DNA
leukaemia	NN	O	I-DNA
factor	NN	O	I-DNA
)	NN	O	I-DNA
fusion	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
formed	NN	O	O
by	NN	O	O
action	NN	O	O
of	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
17	NN	O	I-DNA
;	NN	O	I-DNA
19	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q22	NN	O	I-DNA
;	NN	O	I-DNA
p13	NN	O	I-DNA
)	NN	O	I-DNA
chromosomal	NN	O	O
translocation	NN	O	O
,	NN	O	O
drives	NN	O	O
the	NN	O	O
leukaemic	NN	O	O
transformation	NN	O	O
of	NN	O	O
early	NN	O	O
B-cell	NN	O	B-cell_type
precursors	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
this	NN	O	O
activity	NN	O	O
remains	NN	O	O
unknown	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
leukaemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
carrying	NN	O	O
the	NN	O	O
translocation	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
17	NN	O	I-DNA
;	NN	O	I-DNA
19	NN	O	I-DNA
)	NN	O	I-DNA
rapidly	NN	O	O
died	NN	O	O
by	NN	O	O
apoptosis	NN	O	O
when	NN	O	O
programmed	NN	O	O
to	NN	O	O
express	NN	O	O
a	NN	O	O
dominant-negative	NN	O	O
suppressor	NN	O	O
of	NN	O	O
the	NN	O	O
fusion	NN	O	O
protein	NN	O	O
E2A-HLF	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
chimaeric	NN	O	B-protein
oncoprotein	NN	O	I-protein
probably	NN	O	O
affects	NN	O	O
cell	NN	O	O
survival	NN	O	O
rather	NN	O	O
than	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
when	NN	O	O
introduced	NN	O	O
into	NN	O	O
murine	NN	O	B-cell_line
pro-B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
oncogenic	NN	O	B-protein
E2A-HLF	NN	O	I-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
reversed	NN	O	O
both	NN	O	O
interleukin-3-dependent	NN	O	O
and	NN	O	O
p53-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
close	NN	O	O
homology	NN	O	O
of	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
region/leucine	NN	O	I-protein
zipper	NN	O	I-protein
(	NN	O	O
bZIP	NN	O	B-protein
)	NN	O	O
DNA-binding	NN	O	O
and	NN	O	O
dimerization	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
HLF	NN	O	B-protein
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
CES-2	NN	O	B-protein
cell-death	NN	O	I-protein
specification	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
Caenorhabditis	NN	O	O
elegans	NN	O	O
suggests	NN	O	O
a	NN	O	O
model	NN	O	O
of	NN	O	O
leukaemogenesis	NN	O	O
in	NN	O	O
which	NN	O	O
E2A-HLF	NN	O	B-protein
blocks	NN	O	O
an	NN	O	O
early	NN	O	O
step	NN	O	O
within	NN	O	O
an	NN	O	O
evolutionarily	NN	O	O
conserved	NN	O	O
cell-death	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
early	NN	O	B-DNA
growth	NN	O	I-DNA
response	NN	O	I-DNA
gene	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	O
egr-1	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
is	NN	O	O
critical	NN	O	O
to	NN	O	O
determining	NN	O	O
the	NN	O	O
ultimate	NN	O	O
outcome	NN	O	O
of	NN	O	O
exposure	NN	O	O
to	NN	O	O
antigen	NN	O	O
.	NN	O	O

The	NN	O	O
importance	NN	O	O
of	NN	O	O
many	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
relates	NN	O	O
to	NN	O	O
the	NN	O	O
role	NN	O	O
their	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
products	NN	O	I-protein
play	NN	O	O
in	NN	O	O
dictating	NN	O	O
patterns	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
downstream	NN	O	O
,	NN	O	O
function-related	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Evidence	NN	O	O
from	NN	O	O
several	NN	O	O
systems	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
egr-1	NN	O	B-DNA
may	NN	O	O
be	NN	O	O
of	NN	O	O
particular	NN	O	O
importance	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
the	NN	O	O
egr-1	NN	O	B-DNA
promoter	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
highly	NN	O	O
responsive	NN	O	O
to	NN	O	O
the	NN	O	O
diverse	NN	O	O
biochemical	NN	O	O
signals	NN	O	O
generated	NN	O	O
by	NN	O	O
antigen	NN	O	O
and	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
for	NN	O	O
egr-1	NN	O	B-DNA
in	NN	O	O
determining	NN	O	O
the	NN	O	O
differentiation	NN	O	O
pathway	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
cell	NN	O	I-cell_type
precursors	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
recently	NN	O	O
elaborated	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
potential	NN	O	O
targets	NN	O	O
of	NN	O	O
regulation	NN	O	O
by	NN	O	O
the	NN	O	O
zinc-finger	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
egr-1	NN	O	B-DNA
include	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
,	NN	O	I-DNA
CD44	NN	O	I-DNA
,	NN	O	I-DNA
ICAM-1	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
tumor	NN	O	I-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
egr-1	NN	O	B-DNA
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
will	NN	O	O
be	NN	O	O
discussed	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
these	NN	O	O
recent	NN	O	O
studies	NN	O	O
.	NN	O	O

-DOCSTART-	O

Peptide	NN	O	O
vaccination	NN	O	O
can	NN	O	O
lead	NN	O	O
to	NN	O	O
enhanced	NN	O	O
tumor	NN	O	O
growth	NN	O	O
through	NN	O	O
specific	NN	O	O
T-cell	NN	O	O
tolerance	NN	O	O
induction	NN	O	O
.	NN	O	O

Vaccination	NN	O	O
with	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
representing	NN	O	O
cytotoxic	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
(	NN	O	O
CTL	NN	O	O
)	NN	O	O
epitopes	NN	O	O
can	NN	O	O
lead	NN	O	O
to	NN	O	O
a	NN	O	O
protective	NN	O	O
CTL-mediated	NN	O	O
immunity	NN	O	O
against	NN	O	O
tumors	NN	O	O
or	NN	O	O
viruses	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
that	NN	O	O
vaccination	NN	O	O
with	NN	O	O
a	NN	O	O
CTL	NN	O	O
epitope	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
adenovirus	NN	O	I-DNA
type	NN	O	I-DNA
5	NN	O	I-DNA
E1A-region	NN	O	I-DNA
(	NN	O	O
Ad5E1A234-243	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
which	NN	O	O
can	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
tumor-eradicating	NN	O	B-cell_type
CTL	NN	O	I-cell_type
,	NN	O	O
enhances	NN	O	O
rather	NN	O	O
than	NN	O	O
inhibits	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
Ad5E1A-expressing	NN	O	O
tumors	NN	O	O
.	NN	O	O

This	NN	O	O
adverse	NN	O	O
effect	NN	O	O
of	NN	O	O
peptide	NN	O	O
vaccination	NN	O	O
was	NN	O	O
rapidly	NN	O	O
evoked	NN	O	O
,	NN	O	O
required	NN	O	O
low	NN	O	O
doses	NN	O	O
of	NN	O	O
peptide	NN	O	O
(	NN	O	O
10	NN	O	O
micrograms	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
achieved	NN	O	O
by	NN	O	O
a	NN	O	O
mode	NN	O	O
of	NN	O	O
peptide	NN	O	O
delivery	NN	O	O
that	NN	O	O
induces	NN	O	O
protective	NN	O	O
T-cell-mediated	NN	O	O
immunity	NN	O	O
in	NN	O	O
other	NN	O	O
models	NN	O	O
.	NN	O	O

Ad5E1A	NN	O	B-protein
-specific	NN	O	O
CTL	NN	O	O
activity	NN	O	O
could	NN	O	O
no	NN	O	O
longer	NN	O	O
be	NN	O	O
isolated	NN	O	O
from	NN	O	O
mice	NN	O	O
after	NN	O	O
injection	NN	O	O
of	NN	O	O
Ad5E1A-peptide	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
tolerization	NN	O	O
of	NN	O	O
Ad5E1A	NN	O	B-protein
-specific	NN	O	O
CTL	NN	O	O
activity	NN	O	O
causes	NN	O	O
the	NN	O	O
enhanced	NN	O	O
tumor	NN	O	O
outgrowth	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
peptide	NN	O	O
vaccination	NN	O	O
,	NN	O	O
immunization	NN	O	O
with	NN	O	O
adenovirus	NN	O	O
,	NN	O	O
expressing	NN	O	O
Ad5E1A	NN	O	B-protein
,	NN	O	O
induced	NN	O	O
Ad5E1A	NN	O	B-protein
-specific	NN	O	O
immunity	NN	O	O
and	NN	O	O
prevented	NN	O	O
the	NN	O	O
outgrowth	NN	O	O
of	NN	O	O
Ad5E1A	NN	O	B-protein
-expressing	NN	O	O
tumors	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
immunization	NN	O	O
with	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
can	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
elimination	NN	O	O
of	NN	O	O
anti-tumor	NN	O	O
CTL	NN	O	O
responses	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
design	NN	O	O
of	NN	O	O
safe	NN	O	O
peptide-based	NN	O	O
vaccines	NN	O	O
against	NN	O	O
tumors	NN	O	O
,	NN	O	O
allogeneic	NN	O	O
organ	NN	O	O
transplants	NN	O	O
,	NN	O	O
and	NN	O	O
T-cell-mediated	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
NGFI-B/nur77	NN	O	B-protein
family	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
T-cell	NN	O	O
apoptosis	NN	O	O
]	NN	O	O

NGFI-B/nur77	NN	O	B-protein
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	B-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
.	NN	O	O

NGFI-B/nur77	NN	O	B-protein
and	NN	O	O
its	NN	O	O
related	NN	O	O
genes	NN	O	O
constitute	NN	O	O
a	NN	O	O
family	NN	O	O
and	NN	O	O
the	NN	O	O
NGFI-B/nur77	NN	O	B-protein
family	NN	O	I-protein
consists	NN	O	O
of	NN	O	O
three	NN	O	O
subtypes	NN	O	O
,	NN	O	O
named	NN	O	O
nur77	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
nur77	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
nur77	NN	O	B-protein
gamma	NN	O	I-protein
.	NN	O	O

We	NN	O	O
cloned	NN	O	O
human	NN	O	O
nur77	NN	O	B-DNA
beta	NN	O	I-DNA
cDNA	NN	O	I-DNA
,	NN	O	O
called	NN	O	O
TINUR	NN	O	B-DNA
.	NN	O	O

Although	NN	O	O
NGFI-B/nur77	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
TCR-mediated	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
T-cell	NN	O	B-protein
hybridomas	NN	O	I-protein
,	NN	O	O
the	NN	O	O
reports	NN	O	O
on	NN	O	O
nur77	NN	O	O
knock-out	NN	O	O
mice	NN	O	O
and	NN	O	O
nur77	NN	O	O
dominant	NN	O	O
negative	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
suggest	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
functional	NN	O	O
redundancy	NN	O	O
among	NN	O	O
NGFI-B/nur77	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

NGFI-B/nur77	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
the	NN	O	O
response	NN	O	B-DNA
element	NN	O	I-DNA
by	NN	O	O
monomer	NN	O	O
or	NN	O	O
heterodimer	NN	O	O
with	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RXR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Assuming	NN	O	O
that	NN	O	O
9-cis-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
9-cis-RA	NN	O	O
)	NN	O	O
inhibits	NN	O	O
TCR-mediated	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
nur77	NN	O	B-protein
may	NN	O	O
cause	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
monomer	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
9-cis-RA	NN	O	O
and	NN	O	O
may	NN	O	O
inhibit	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
heterodimer	NN	O	O
with	NN	O	O
RXR	NN	O	B-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
9-cis-RA	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoids	NN	O	O
and	NN	O	O
interferon-alpha	NN	O	B-protein
in	NN	O	O
the	NN	O	O
acquired	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Some	NN	O	O
patients	NN	O	O
with	NN	O	O
acquired	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
(	NN	O	O
AIDS	NN	O	O
)	NN	O	O
develop	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
characterized	NN	O	O
by	NN	O	O
low	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
for	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
,	NN	O	O
cells	NN	O	O
and	NN	O	O
high	NN	O	O
values	NN	O	O
of	NN	O	O
ACTH	NN	O	O
and	NN	O	O
cortisol	NN	O	O
.	NN	O	O

As	NN	O	O
glucocorticoids	NN	O	O
regulate	NN	O	O
interferon-alpha	NN	O	B-protein
(	NN	O	O
IFN	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
production	NN	O	O
,	NN	O	O
we	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
IFN	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
produced	NN	O	O
predominantly	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
in	NN	O	O
AIDS	NN	O	O
,	NN	O	O
should	NN	O	O
be	NN	O	O
increased	NN	O	O
in	NN	O	O
cortisol-resistant	NN	O	O
AIDS	NN	O	O
,	NN	O	O
attributing	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
cortisol	NN	O	O
inhibition	NN	O	O
to	NN	O	O
IFN	NN	O	B-protein
alpha	NN	O	I-protein
production	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
characteristics	NN	O	O
on	NN	O	O
monocytes	NN	O	B-cell_type
by	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
and	NN	O	O
measured	NN	O	O
IFN	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
cortisol	NN	O	O
,	NN	O	O
and	NN	O	O
ACTH	NN	O	O
in	NN	O	O
AIDS	NN	O	O
patients	NN	O	O
with	NN	O	O
(	NN	O	O
AIDS-GR	NN	O	O
)	NN	O	O
or	NN	O	O
without	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
(	NN	O	O
AIDS-C	NN	O	O
)	NN	O	O
and	NN	O	O
controls	NN	O	O
(	NN	O	O
C	NN	O	O
)	NN	O	O
.	NN	O	O

Monocytes	NN	O	O
of	NN	O	O
AIDS-GR	NN	O	O
patients	NN	O	O
had	NN	O	O
a	NN	O	O
receptor	NN	O	O
Kd	NN	O	O
of	NN	O	O
10.5	NN	O	O
+/-	NN	O	O
4.2	NN	O	O
nmol/L	NN	O	O
that	NN	O	O
was	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
AIDS-C	NN	O	O
group	NN	O	O
(	NN	O	O
2.9	NN	O	O
+/-	NN	O	O
0.8	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
and	NN	O	O
normal	NN	O	O
subjects	NN	O	O
(	NN	O	O
2.0	NN	O	O
+/-	NN	O	O
0.8	NN	O	O
nmol/L	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

IFN	NN	O	B-protein
alpha	NN	O	I-protein
levels	NN	O	O
were	NN	O	O
increased	NN	O	O
in	NN	O	O
the	NN	O	O
AIDS-GR	NN	O	O
group	NN	O	O
(	NN	O	O
17	NN	O	O
+/-	NN	O	O
6	NN	O	O
vs	NN	O	O
.	NN	O	O

4	NN	O	O
+/-	NN	O	O
1	NN	O	O
U/mL	NN	O	O
in	NN	O	O
the	NN	O	O
AIDS-C	NN	O	O
group	NN	O	O
and	NN	O	O
2	NN	O	O
+/-	NN	O	O
0.5	NN	O	O
U/mL	NN	O	O
in	NN	O	O
the	NN	O	O
C	NN	O	O
group	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

Correlations	NN	O	O
were	NN	O	O
found	NN	O	O
between	NN	O	O
plasma	NN	O	B-protein
IFN	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
receptor	NN	O	O
Kd	NN	O	O
on	NN	O	O
monocytes	NN	O	B-cell_type
of	NN	O	O
AIDS-GR	NN	O	O
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.77	NN	O	O
)	NN	O	O
and	NN	O	O
between	NN	O	O
IFN	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
group	NN	O	O
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.74	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
poly	NN	O	B-protein
(	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
-poly	NN	O	I-protein
(	NN	O	I-protein
C	NN	O	I-protein
)	NN	O	I-protein
-induced	NN	O	I-protein
IFN	NN	O	I-protein
alpha	NN	O	I-protein
production	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
the	NN	O	O
C	NN	O	O
and	NN	O	O
AIDS-C	NN	O	O
groups	NN	O	O
(	NN	O	O
approximately	NN	O	O
80	NN	O	O
%	NN	O	O
inhibition	NN	O	O
in	NN	O	O
both	NN	O	O
groups	NN	O	O
)	NN	O	O
;	NN	O	O
the	NN	O	O
effect	NN	O	O
was	NN	O	O
reversed	NN	O	O
by	NN	O	O
the	NN	O	O
receptor	NN	O	O
antagonist	NN	O	O
RU-38486	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
glucocorticoids	NN	O	O
failed	NN	O	O
to	NN	O	O
inhibit	NN	O	O
IFNalpha	NN	O	B-protein
production	NN	O	O
from	NN	O	O
AIDS-GR	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
(	NN	O	O
approximately	NN	O	O
20	NN	O	O
%	NN	O	O
inhibition	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
elevated	NN	O	O
IFN	NN	O	B-protein
alpha	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
AIDS-GR	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
cortisol	NN	O	O
on	NN	O	O
IFN	NN	O	B-protein
alpha	NN	O	I-protein
production	NN	O	O
due	NN	O	O
to	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type

-DOCSTART-	O

JNK	NN	O	B-protein
(	NN	O	O
c-Jun	NN	O	B-protein
NH2-terminal	NN	O	I-protein
kinase	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
antioxidants	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

AP-1	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
behave	NN	O	O
as	NN	O	O
a	NN	O	O
redox-sensitive	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
can	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
both	NN	O	O
oxidant	NN	O	O
and	NN	O	O
antioxidant	NN	O	O
stimuli	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
by	NN	O	O
antioxidants	NN	O	O
are	NN	O	O
largely	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
structurally	NN	O	O
unrelated	NN	O	O
antioxidant	NN	O	O
agents	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
,	NN	O	O
butylated	NN	O	O
hydroxyanisole	NN	O	O
,	NN	O	O
and	NN	O	O
Nacetylcysteine	NN	O	O
activated	NN	O	O
JNK	NN	O	B-protein
(	NN	O	O
c-Jun	NN	O	B-protein
NH2-terminal	NN	O	I-protein
kinase	NN	O	I-protein
)	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
activation	NN	O	O
differed	NN	O	O
substantially	NN	O	O
from	NN	O	O
that	NN	O	O
mediated	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
or	NN	O	O
produced	NN	O	O
by	NN	O	O
costimulation	NN	O	O
with	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor-CD3	NN	O	I-protein
complex	NN	O	I-protein
and	NN	O	O
to	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
JNK	NN	O	B-protein
by	NN	O	O
classical	NN	O	O
T	NN	O	O
cell	NN	O	O
stimuli	NN	O	O
was	NN	O	O
transient	NN	O	O
,	NN	O	O
whereas	NN	O	O
that	NN	O	O
mediated	NN	O	O
by	NN	O	O
PDTC	NN	O	O
and	NN	O	O
butylated	NN	O	O
hydroxyanisole	NN	O	O
(	NN	O	O
but	NN	O	O
not	NN	O	O
N-acetylcysteine	NN	O	O
)	NN	O	O
was	NN	O	O
sustained	NN	O	O
.	NN	O	O

The	NN	O	O
kinetics	NN	O	O
of	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
which	NN	O	O
was	NN	O	O
transient	NN	O	O
after	NN	O	O
stimulation	NN	O	O
with	NN	O	O
PMA	NN	O	O
plus	NN	O	O
ionophore	NN	O	O
and	NN	O	O
prolonged	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
PDTC	NN	O	O
,	NN	O	O
which	NN	O	O
also	NN	O	O
transiently	NN	O	O
induced	NN	O	O
c-fos	NN	O	B-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
PMA	NN	O	O
plus	NN	O	O
ionophore	NN	O	O
was	NN	O	O
sensitive	NN	O	O
to	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
involving	NN	O	O
Ca2+	NN	O	O
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
,	NN	O	O
and	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
which	NN	O	O
failed	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
activation	NN	O	O
mediated	NN	O	O
by	NN	O	O
PDTC	NN	O	O
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
trans-dominant	NN	O	B-DNA
negative	NN	O	I-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
of	NN	O	O
ras	NN	O	B-DNA
and	NN	O	O
raf	NN	O	B-DNA
,	NN	O	O
together	NN	O	O
with	NN	O	O
AP-1-dependent	NN	O	B-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
using	NN	O	O
anti-ERK	NN	O	B-protein
(	NN	O	I-protein
extracellular	NN	O	I-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
)	NN	O	I-protein
antibodies	NN	O	I-protein
,	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
Ras/Raf/ERK	NN	O	O
pathway	NN	O	O
did	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
combined	NN	O	O
treatment	NN	O	O
with	NN	O	O
PDTC	NN	O	O
and	NN	O	O
PMA	NN	O	O
,	NN	O	O
two	NN	O	O
agents	NN	O	O
that	NN	O	O
synergize	NN	O	O
on	NN	O	O
AP-1	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
persistent	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
ERK-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
identify	NN	O	O
JNK	NN	O	B-protein
as	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
antioxidant	NN	O	O
agents	NN	O	O
which	NN	O	O
can	NN	O	O
be	NN	O	O
regulated	NN	O	O
differentially	NN	O	O
under	NN	O	O
oxidant	NN	O	O
and	NN	O	O
antioxidant	NN	O	O
conditions	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
age-related	NN	O	O
lymphocyte	NN	O	O
dysfunction	NN	O	O
]	NN	O	O

Aging	NN	O	O
is	NN	O	O
classically	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
dysregulation	NN	O	O
of	NN	O	O
the	NN	O	O
immunologic	NN	O	O
machinery	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
consequence	NN	O	O
,	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
developed	NN	O	O
in	NN	O	O
senescent	NN	O	O
organisms	NN	O	O
is	NN	O	O
usually	NN	O	O
inappropriate	NN	O	O
,	NN	O	O
often	NN	O	O
inefficient	NN	O	O
,	NN	O	O
sometimes	NN	O	O
aberrant	NN	O	O
,	NN	O	O
and	NN	O	O
potentially	NN	O	O
detrimental	NN	O	O
.	NN	O	O

The	NN	O	O
age-associated	NN	O	O
immune	NN	O	O
dysfunction	NN	O	O
may	NN	O	O
be	NN	O	O
implicated	NN	O	O
to	NN	O	O
some	NN	O	O
degree	NN	O	O
in	NN	O	O
the	NN	O	O
extreme	NN	O	O
susceptibility	NN	O	O
of	NN	O	O
the	NN	O	O
elderly	NN	O	O
to	NN	O	O
infection	NN	O	O
and	NN	O	O
neoplasia	NN	O	O
and	NN	O	O
may	NN	O	O
even	NN	O	O
participate	NN	O	O
in	NN	O	O
various	NN	O	O
aspects	NN	O	O
of	NN	O	O
senescence	NN	O	O
.	NN	O	O

The	NN	O	O
current	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
immunosenescence	NN	O	O
is	NN	O	O
still	NN	O	O
fragmentary	NN	O	O
.	NN	O	O

The	NN	O	O
most	NN	O	O
extensively	NN	O	O
studied	NN	O	O
phenomenon	NN	O	O
is	NN	O	O
the	NN	O	O
progressive	NN	O	O
decline	NN	O	O
in	NN	O	O
the	NN	O	O
proliferative	NN	O	O
capacities	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
aging	NN	O	O
.	NN	O	O

The	NN	O	O
loss	NN	O	O
of	NN	O	O
proliferative	NN	O	O
potential	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
antigenic	NN	O	O
challenge	NN	O	O
is	NN	O	O
a	NN	O	O
characteristic	NN	O	O
feature	NN	O	O
of	NN	O	O
immune	NN	O	O
senescence	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
directly	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
emergence	NN	O	O
of	NN	O	O
the	NN	O	O
age-related	NN	O	O
immune	NN	O	O
deficiency	NN	O	O
.	NN	O	O

The	NN	O	O
purpose	NN	O	O
of	NN	O	O
this	NN	O	O
review	NN	O	O
is	NN	O	O
to	NN	O	O
show	NN	O	O
how	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
various	NN	O	O
biochemical	NN	O	O
lesions	NN	O	O
with	NN	O	O
advancing	NN	O	O
age	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
failure	NN	O	O
of	NN	O	O
a	NN	O	O
critical	NN	O	O
cell	NN	O	O
function	NN	O	O
,	NN	O	O
namely	NN	O	O
the	NN	O	O
activation-induced	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
.	NN	O	O

The	NN	O	O
biochemical	NN	O	O
modifications	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
defect	NN	O	O
in	NN	O	O
transduction	NN	O	O
and	NN	O	O
execution	NN	O	O
of	NN	O	O
the	NN	O	O
proliferative	NN	O	O
signal	NN	O	O
are	NN	O	O
analyzed	NN	O	O
as	NN	O	O
a	NN	O	O
function	NN	O	O
of	NN	O	O
age	NN	O	O
.	NN	O	O

The	NN	O	O
multiple	NN	O	O
alterations	NN	O	O
observed	NN	O	O
on	NN	O	O
the	NN	O	O
various	NN	O	O
biochemical	NN	O	O
pathways	NN	O	O
may	NN	O	O
appear	NN	O	O
as	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
a	NN	O	O
unique	NN	O	O
deleterious	NN	O	O
mechanism	NN	O	O
more	NN	O	O
fundamentally	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
senescence	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
inability	NN	O	O
to	NN	O	O
cope	NN	O	O
with	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
phosphorylated	NN	O	O
CREB	NN	O	O
proteins	NN	O	O
during	NN	O	O
B-cell	NN	O	O
activation	NN	O	O
and	NN	O	O
rescue	NN	O	O
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Engagement	NN	O	O
of	NN	O	O
surface	NN	O	O
immunoglobulin	NN	O	O
on	NN	O	O
mature	NN	O	O
B	NN	O	O
cells	NN	O	O
leads	NN	O	O
to	NN	O	O
rescue	NN	O	O
from	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
to	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Levels	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
increase	NN	O	O
with	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
surface	NN	O	B-protein
immunoglobulin	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
located	NN	O	O
the	NN	O	O
major	NN	O	O
positive	NN	O	O
regulatory	NN	O	O
region	NN	O	O
for	NN	O	O
control	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
positive	NN	O	O
region	NN	O	O
can	NN	O	O
be	NN	O	O
divided	NN	O	O
into	NN	O	O
an	NN	O	O
upstream	NN	O	B-DNA
and	NN	O	I-DNA
a	NN	O	I-DNA
downstream	NN	O	I-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
downstream	NN	O	O
regulatory	NN	O	O
region	NN	O	O
contains	NN	O	O
a	NN	O	O
cyclic	NN	O	B-DNA
AMP-responsive	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CRE	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
by	NN	O	O
antibody	NN	O	O
supershift	NN	O	O
experiments	NN	O	O
and	NN	O	O
UV	NN	O	O
cross-linking	NN	O	O
followed	NN	O	O
by	NN	O	O
denaturing	NN	O	O
polyacrylamide	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
that	NN	O	O
both	NN	O	O
CREB	NN	O	O
and	NN	O	O
ATF	NN	O	O
family	NN	O	O
members	NN	O	O
bind	NN	O	O
to	NN	O	O
this	NN	O	O
region	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Mutations	NN	O	O
of	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
that	NN	O	O
result	NN	O	O
in	NN	O	O
loss	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
binding	NN	O	O
also	NN	O	O
lead	NN	O	O
to	NN	O	O
loss	NN	O	O
of	NN	O	O
functional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
bcl-2	NN	O	O
promoter	NN	O	O
in	NN	O	O
transient-transfection	NN	O	O
assays	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
active	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
bcl-2	NN	O	B-DNA
promoter	NN	O	I-DNA
implies	NN	O	O
that	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
linked	NN	O	O
to	NN	O	O
a	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
the	NN	O	O
mature	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
BAL-17	NN	O	I-cell_line
with	NN	O	O
either	NN	O	O
anti-immunoglobulin	NN	O	O
M	NN	O	O
or	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
leads	NN	O	O
to	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
that	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
the	NN	O	O
more	NN	O	O
immature	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
Ramos	NN	O	B-cell_line
,	NN	O	O
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
rescues	NN	O	O
the	NN	O	O
cells	NN	O	O
from	NN	O	O
calcium-dependent	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

bcl-2	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
increased	NN	O	O
following	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
treatment	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
increased	NN	O	O
expression	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
stimuli	NN	O	O
result	NN	O	O
in	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
at	NN	O	O
serine	NN	O	O
133	NN	O	O
.	NN	O	O

The	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
that	NN	O	O
results	NN	O	O
in	NN	O	O
activation	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
rather	NN	O	O
than	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
necessary	NN	O	O
,	NN	O	O
optimal	NN	O	O
induction	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
requires	NN	O	O
participation	NN	O	O
of	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
alters	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
bcl-2	NN	O	B-DNA
promoter	NN	O	I-DNA
appears	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
during	NN	O	O
the	NN	O	O
rescue	NN	O	O
of	NN	O	O
immature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
possible	NN	O	O
that	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
particularly	NN	O	O
those	NN	O	O
in	NN	O	O
which	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
myeloid	NN	O	I-DNA
cell	NN	O	I-DNA
nuclear	NN	O	I-DNA
differentiation	NN	O	I-DNA
antigen	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

MNDA	NN	O	B-protein
(	NN	O	O
myeloid	NN	O	B-protein
cell	NN	O	I-protein
nuclear	NN	O	I-protein
differentiation	NN	O	I-protein
antigen	NN	O	I-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
interferon	NN	O	B-protein
alpha	NN	O	I-protein
regulated	NN	O	I-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
expressed	NN	O	O
only	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_type
myelomonocytic	NN	O	I-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
identify	NN	O	O
mechanisms	NN	O	O
responsible	NN	O	O
for	NN	O	O
this	NN	O	O
lineage-specific	NN	O	O
and	NN	O	O
interferon-regulated	NN	O	O
expression	NN	O	O
,	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
characterized	NN	O	O
.	NN	O	O

Two	NN	O	O
interferon-stimulated	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
(	NN	O	O
ISRE	NN	O	B-DNA
)	NN	O	O
flank	NN	O	O
a	NN	O	O
multiple	NN	O	O
transcription	NN	O	O
start	NN	O	O
site	NN	O	O
region	NN	O	O
identifying	NN	O	O
MNDA	NN	O	B-protein
as	NN	O	O
a	NN	O	O
TATA-less	NN	O	B-DNA
interferon-regulated	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Other	NN	O	O
DNA	NN	O	B-DNA
elements	NN	O	I-DNA
present	NN	O	O
include	NN	O	O
a	NN	O	O
cluster	NN	O	O
of	NN	O	O
Myb	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
several	NN	O	O
Ets	NN	O	B-DNA
,	NN	O	O
an	NN	O	O
Ets	NN	O	B-DNA
related	NN	O	I-DNA
PU.1	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
an	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
located	NN	O	O
within	NN	O	O
600	NN	O	O
bp	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
DNA	NN	O	O
methylation	NN	O	O
was	NN	O	O
revealed	NN	O	O
as	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
possible	NN	O	O
factors	NN	O	O
in	NN	O	O
establishing	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
sequence	NN	O	I-DNA
has	NN	O	O
promoter	NN	O	O
activity	NN	O	O
which	NN	O	O
is	NN	O	O
elevated	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
mechanisms	NN	O	O
similar	NN	O	O
to	NN	O	O
other	NN	O	O
myelomonocytic	NN	O	B-DNA
cell	NN	O	I-DNA
specific	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
genes	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Abnormality	NN	O	O
of	NN	O	O
Oct-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
Sjogren	NN	O	O
's	NN	O	O
syndrome	NN	O	O
patients	NN	O	O
.	NN	O	O

Primary	NN	O	O
Sjogren	NN	O	O
's	NN	O	O
syndrome	NN	O	O
(	NN	O	O
SS	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
autoimmune	NN	O	O
rheumatic	NN	O	O
disease	NN	O	O
characterized	NN	O	O
by	NN	O	O
T	NN	O	O
cell	NN	O	O
hypoactivity	NN	O	O
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
diminished	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
to	NN	O	O
activation	NN	O	O
signals	NN	O	O
,	NN	O	O
we	NN	O	O
measured	NN	O	O
nucleoprotein	NN	O	O
DNA-binding	NN	O	O
activities	NN	O	O
regulating	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
using	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
9/19	NN	O	O
SS	NN	O	O
patients	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
defective	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
bind	NN	O	O
an	NN	O	O
october	NN	O	B-protein
sequence	NN	O	I-protein
(	NN	O	O
Oct-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

This	NN	O	O
Oct-1	NN	O	B-protein
-binding	NN	O	O
phenotype	NN	O	O
remained	NN	O	O
stable	NN	O	O
in	NN	O	O
culture	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
3	NN	O	O
days	NN	O	O
prior	NN	O	O
to	NN	O	O
activation	NN	O	O
.	NN	O	O

This	NN	O	O
abnormality	NN	O	O
was	NN	O	O
not	NN	O	O
seen	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
nor	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
,	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
SS	NN	O	O
accompanied	NN	O	O
by	NN	O	O
RA	NN	O	O
.	NN	O	O

The	NN	O	O
SS	NN	O	O
Oct-1	NN	O	B-protein
DNA-binding	NN	O	O
abnormality	NN	O	O
correlated	NN	O	O
significantly	NN	O	O
with	NN	O	O
an	NN	O	O
inability	NN	O	O
of	NN	O	O
cells	NN	O	O
to	NN	O	O
exit	NN	O	O
the	NN	O	O
Gzero/G1	NN	O	O
cell	NN	O	O
cycle	NN	O	O
phase	NN	O	O
when	NN	O	O
stimulated	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
nucleoprotein	NN	O	O
extracts	NN	O	O
showing	NN	O	O
decreased	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
had	NN	O	O
normal	NN	O	O
amounts	NN	O	O
of	NN	O	O
Oct-1	NN	O	B-protein
proteins	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
immunoprecipitation	NN	O	O
,	NN	O	O
implying	NN	O	O
a	NN	O	O
functional	NN	O	O
defect	NN	O	O
in	NN	O	O
the	NN	O	O
Oct-1	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
defective	NN	O	O
DNA	NN	O	O
binding	NN	O	O
was	NN	O	O
corrected	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
acid	NN	O	O
phosphatase	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
isoform	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
(	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
)	NN	O	O
gene	NN	O	O
family	NN	O	O
member	NN	O	O
NFATc	NN	O	B-protein
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
J	NN	O	O
Biol	NN	O	O
Chem	NN	O	O
1996	NN	O	O
Dec	NN	O	O
27	NN	O	O
;	NN	O	O
271	NN	O	O
(	NN	O	O
52	NN	O	O
)	NN	O	O
:	NN	O	O
33705	NN	O	O
]	NN	O	O

The	NN	O	O
cyclosporin	NN	O	B-protein
A	NN	O	I-protein
(	NN	O	I-protein
CsA	NN	O	I-protein
)	NN	O	I-protein
/FK506-sensitive	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
NFAT	NN	O	B-protein
has	NN	O	O
been	NN	O	O
recently	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
inducible	NN	O	O
in	NN	O	O
several	NN	O	O
non-T	NN	O	B-cell_type
immune	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
members	NN	O	I-DNA
characterized	NN	O	O
to	NN	O	O
date	NN	O	O
have	NN	O	O
been	NN	O	O
isolated	NN	O	O
only	NN	O	O
from	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
further	NN	O	O
characterize	NN	O	O
NFAT	NN	O	B-protein
function	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
specificity	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
we	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
isolation	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
Raji	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
new	NN	O	O
isoform	NN	O	O
,	NN	O	O
NFATc.beta	NN	O	B-protein
,	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
gene	NN	O	I-protein
family	NN	O	I-protein
member	NN	O	I-protein
NFATc	NN	O	B-protein
(	NN	O	O
designated	NN	O	O
here	NN	O	O
NFATc.alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
amino	NN	O	B-protein
acid	NN	O	I-protein
sequence	NN	O	I-protein
of	NN	O	O
NFATc.beta	NN	O	B-protein
differs	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
NFATc	NN	O	B-protein
.	NN	O	O

alpha	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	O
NH2-terminal	NN	O	B-protein
29	NN	O	I-protein
residues	NN	O	I-protein
and	NN	O	O
contains	NN	O	O
an	NN	O	O
additional	NN	O	O
region	NN	O	O
of	NN	O	O
142	NN	O	B-protein
residues	NN	O	I-protein
at	NN	O	O
the	NN	O	O
COOH	NN	O	B-protein
terminus	NN	O	I-protein
.	NN	O	O

Northern	NN	O	O
analysis	NN	O	O
using	NN	O	O
a	NN	O	O
probe	NN	O	O
encompassing	NN	O	O
a	NN	O	O
common	NN	O	O
region	NN	O	O
of	NN	O	O
both	NN	O	O
isoforms	NN	O	O
showed	NN	O	O
two	NN	O	O
mRNA	NN	O	B-RNA
species	NN	O	I-RNA
of	NN	O	O
2.7	NN	O	B-RNA
and	NN	O	I-RNA
4.5	NN	O	I-RNA
kilobase	NN	O	I-RNA
pairs	NN	O	I-RNA
,	NN	O	O
while	NN	O	O
an	NN	O	O
NFATc.beta-specific	NN	O	B-DNA
probe	NN	O	I-DNA
detected	NN	O	O
only	NN	O	O
the	NN	O	O
4.5-kilobase	NN	O	B-RNA
pair	NN	O	I-RNA
mRNA	NN	O	I-RNA
which	NN	O	O
was	NN	O	O
preferentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
spleen	NN	O	O
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
of	NN	O	O
NFATc.beta	NN	O	B-protein
was	NN	O	O
capable	NN	O	O
of	NN	O	O
activating	NN	O	O
an	NN	O	O
interleukin-2	NN	O	B-DNA
NFAT-driven	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
stimulated	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
a	NN	O	O
CsA-sensitive	NN	O	O
manner	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
NFATc.beta	NN	O	B-protein
neither	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
kappa3	NN	O	B-DNA
element	NN	O	I-DNA
(	NN	O	O
an	NN	O	O
NFAT-binding	NN	O	B-DNA
site	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor-alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
nor	NN	O	O
activated	NN	O	O
the	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor-alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
cotransfection	NN	O	O
assays	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
different	NN	O	O
members	NN	O	O
or	NN	O	O
isoforms	NN	O	O
of	NN	O	O
NFAT	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	O
may	NN	O	O
regulate	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
different	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
interferon-gamma	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Interferon-gamma	NN	O	O
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
interferon	NN	O	I-protein
,	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
immunoregulatory	NN	O	O
gene	NN	O	O
that	NN	O	O
has	NN	O	O
multiple	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
development	NN	O	O
,	NN	O	O
maturation	NN	O	O
,	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

IFN-gamma	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
are	NN	O	O
expressed	NN	O	O
predominantly	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
large	NN	O	B-cell_type
granular	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
IFN-gamma	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
induced/inhibited	NN	O	O
in	NN	O	O
these	NN	O	O
cell	NN	O	O
types	NN	O	O
by	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
extracellular	NN	O	O
signals	NN	O	O
,	NN	O	O
thus	NN	O	O
implicating	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
diverse	NN	O	O
,	NN	O	O
yet	NN	O	O
convergent	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
in	NN	O	O
its	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
,	NN	O	O
I	NN	O	O
describe	NN	O	O
how	NN	O	O
DNA	NN	O	O
methylation	NN	O	O
and	NN	O	O
specific	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
may	NN	O	O
regulate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
extracellular	NN	O	O
signals	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
analysis	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
EBV-positive	NN	O	O
gastric	NN	O	O
carcinoma	NN	O	O
:	NN	O	O
unique	NN	O	O
viral	NN	O	O
latency	NN	O	O
in	NN	O	O
the	NN	O	O
tumour	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
case-oriented	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
association	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
with	NN	O	O
gastric	NN	O	O
carcinoma	NN	O	O
has	NN	O	O
been	NN	O	O
accumulating	NN	O	O
recently	NN	O	O
,	NN	O	O
the	NN	O	O
interaction	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
between	NN	O	O
EBV	NN	O	O
and	NN	O	O
gastric	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
is/are	NN	O	O
largely	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
seven	NN	O	O
EBV-positive	NN	O	O
gastric	NN	O	O
carcinoma	NN	O	O
tissues	NN	O	O
for	NN	O	O
viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
,	NN	O	O
from	NN	O	O
which	NN	O	O
studies	NN	O	O
on	NN	O	O
the	NN	O	O
EBV	NN	O	O
oncogenicity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
will	NN	O	O
benefit	NN	O	O
.	NN	O	O

Reverse	NN	O	O
transcription-PCR	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
all	NN	O	O
seven	NN	O	O
EBV-positive	NN	O	O
tumour	NN	O	O
tissues	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
EBV	NN	O	B-RNA
nuclear	NN	O	I-RNA
antigen	NN	O	I-RNA
(	NN	O	I-RNA
EBNA	NN	O	I-RNA
)	NN	O	I-RNA
1	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
EBNA2	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

The	NN	O	O
EBNA	NN	O	B-protein
transcription	NN	O	O
was	NN	O	O
initiated	NN	O	O
from	NN	O	O
one	NN	O	O
of	NN	O	O
three	NN	O	O
EBNA	NN	O	B-DNA
promoters	NN	O	I-DNA
,	NN	O	O
Qp	NN	O	B-DNA
:	NN	O	O
by	NN	O	O
contrast	NN	O	O
,	NN	O	O
both	NN	O	O
Cp	NN	O	B-DNA
and	NN	O	O
Wp	NN	O	B-DNA
were	NN	O	O
silent	NN	O	O
,	NN	O	O
thus	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
EBNA2	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Latent	NN	O	B-RNA
membrane	NN	O	I-RNA
protein	NN	O	I-RNA
(	NN	O	I-RNA
LMP	NN	O	I-RNA
)	NN	O	I-RNA
2A	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
detected	NN	O	O
in	NN	O	O
three	NN	O	O
of	NN	O	O
seven	NN	O	O
cases	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
neither	NN	O	B-RNA
LMP1	NN	O	I-RNA
nor	NN	O	I-RNA
LMP2B	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
detected	NN	O	O
in	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
tumours	NN	O	O
tested	NN	O	O
.	NN	O	O

Transcripts	NN	O	O
from	NN	O	O
the	NN	O	O
BamHI-A	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
genome	NN	O	I-DNA
were	NN	O	O
detectable	NN	O	O
in	NN	O	O
all	NN	O	O
cases	NN	O	O
.	NN	O	O

BZLF1	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
the	NN	O	O
product	NN	O	O
,	NN	O	O
an	NN	O	O
immediate-early	NN	O	B-DNA
gene	NN	O	I-DNA
for	NN	O	O
EBV	NN	O	O
replication	NN	O	O
,	NN	O	O
was	NN	O	O
not	NN	O	O
expressed	NN	O	O
in	NN	O	O
any	NN	O	O
of	NN	O	O
them	NN	O	O
,	NN	O	O
thereby	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
tumour	NN	O	B-cell_type
cells	NN	O	I-cell_type
carried	NN	O	O
EBV	NN	O	B-DNA
genomes	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
tightly	NN	O	O
latent	NN	O	O
form	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
further	NN	O	O
extended	NN	O	O
our	NN	O	O
previous	NN	O	O
data	NN	O	O
regarding	NN	O	O
EBV	NN	O	O
latency	NN	O	O
in	NN	O	O
gastric	NN	O	B-cell_type
carcinoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
the	NN	O	O
protein	NN	O	O
level	NN	O	O
,	NN	O	O
and	NN	O	O
have	NN	O	O
affirmed	NN	O	O
that	NN	O	O
the	NN	O	O
programme	NN	O	O
of	NN	O	O
viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
tumour	NN	O	O
more	NN	O	O
closely	NN	O	O
resembles	NN	O	O
'latency	NN	O	O
I	NN	O	O
'	NN	O	O
represented	NN	O	O
by	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
than	NN	O	O
'latency	NN	O	O
II	NN	O	O
'	NN	O	O
represented	NN	O	O
by	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
nasopharyngeal	NN	O	O
carcinomas	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
Grb2-binding	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
platelets	NN	O	I-cell_type
activated	NN	O	O
by	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RIIA	NN	O	I-protein
cross-linking	NN	O	O
.	NN	O	O

Glutathione-S-transferase	NN	O	B-protein
(	NN	O	I-protein
GST	NN	O	I-protein
)	NN	O	I-protein
-Grb2	NN	O	I-protein
fusion	NN	O	I-protein
proteins	NN	O	I-protein
have	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
Grb2-binding	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
platelet	NN	O	B-cell_type
activation	NN	O	O
by	NN	O	O
the	NN	O	O
platelet	NN	O	B-protein
low-affinity	NN	O	I-protein
IgG	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RIIA	NN	O	I-protein
.	NN	O	O

Two	NN	O	O
tyrosine	NN	O	B-protein
phosphoproteins	NN	O	I-protein
of	NN	O	O
38	NN	O	O
and	NN	O	O
63	NN	O	O
kD	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
Grb2	NN	O	B-protein
following	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RIIA	NN	O	I-protein
stimulation	NN	O	O
of	NN	O	O
platelets	NN	O	B-cell_type
.	NN	O	O

Both	NN	O	O
are	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
particulate	NN	O	O
fraction	NN	O	O
following	NN	O	O
platelet	NN	O	B-cell_type
activation	NN	O	O
and	NN	O	O
are	NN	O	O
also	NN	O	O
able	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
a	NN	O	O
GST-construct	NN	O	B-protein
containing	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
and	NN	O	I-protein
SH3	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
phospholipase	NN	O	B-protein
C	NN	O	I-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
.	NN	O	O

p38	NN	O	O
also	NN	O	O
forms	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
csk	NN	O	I-protein
in	NN	O	O
stimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
a	NN	O	O
substrate	NN	O	O
for	NN	O	O
the	NN	O	O
kinase	NN	O	B-protein
.	NN	O	O

The	NN	O	O
SH3	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
Grb2	NN	O	B-protein
form	NN	O	O
a	NN	O	O
stable	NN	O	O
complex	NN	O	O
with	NN	O	O
SOS1	NN	O	B-protein
and	NN	O	O
two	NN	O	O
proteins	NN	O	O
of	NN	O	O
75	NN	O	B-protein
kD	NN	O	I-protein
and	NN	O	O
120	NN	O	B-protein
kD	NN	O	I-protein
,	NN	O	O
which	NN	O	O
undergo	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
Fc	NN	O	B-cell_line
gamma	NN	O	I-cell_line
RIIA	NN	O	I-cell_line
stimulated	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
75-kD	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
recognized	NN	O	O
by	NN	O	O
antibodies	NN	O	O
to	NN	O	O
SLP-76	NN	O	B-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
isolated	NN	O	O
from	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
sequenced	NN	O	O
.	NN	O	O

Tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
and	NN	O	O
p63	NN	O	B-protein
is	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
platelets	NN	O	B-cell_type
stimulated	NN	O	O
by	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
-linked	NN	O	O
receptor	NN	O	O
agonist	NN	O	O
collagen	NN	O	B-protein
and	NN	O	O
by	NN	O	O
the	NN	O	O
G	NN	O	B-protein
protein	NN	O	I-protein
-coupled	NN	O	O
receptor	NN	O	O
agonist	NN	O	O
thrombin	NN	O	B-protein
,	NN	O	O
although	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
SLP-76	NN	O	B-protein
is	NN	O	O
only	NN	O	O
observed	NN	O	O
in	NN	O	O
collagen	NN	O	B-protein
-stimulated	NN	O	O
platelets	NN	O	B-cell_type
.	NN	O	O

p38	NN	O	B-protein
and	NN	O	O
p63	NN	O	B-protein
may	NN	O	O
provide	NN	O	O
a	NN	O	O
docking	NN	O	O
site	NN	O	O
for	NN	O	O
Grb2	NN	O	B-protein
,	NN	O	O
thereby	NN	O	O
linking	NN	O	O
Grb2	NN	O	B-protein
SH3-binding	NN	O	I-protein
proteins	NN	O	I-protein
SOS1	NN	O	B-protein
,	NN	O	O
SLP-76	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p120	NN	O	B-protein
to	NN	O	O
downstream	NN	O	O
signalling	NN	O	O
events	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
specific	NN	O	O
interferon	NN	O	B-protein
isotypes	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
from	NN	O	O
normal	NN	O	O
individuals	NN	O	O
and	NN	O	O
in	NN	O	O
promonocytic	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
IFN-alpha5	NN	O	B-protein
and	NN	O	O
IFN-beta	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
different	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
including	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
normal	NN	O	O
individuals	NN	O	O
following	NN	O	O
amplification	NN	O	O
of	NN	O	O
IFN	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
and	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
amplified	NN	O	B-DNA
product	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
activated	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
interferon-induced	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complex	NN	O	I-protein
ISGF3	NN	O	B-protein
was	NN	O	O
also	NN	O	O
detected	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
uninduced	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Culture	NN	O	O
supernatants	NN	O	O
from	NN	O	O
uninduced	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
also	NN	O	O
found	NN	O	O
to	NN	O	O
activate	NN	O	O
an	NN	O	O
ISRE	NN	O	B-DNA
cloned	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
IFN	NN	O	I-protein
activity	NN	O	O
equivalent	NN	O	O
to	NN	O	O
approximately	NN	O	O
0.3	NN	O	O
to	NN	O	O
0.5	NN	O	O
IU/mL	NN	O	O
.	NN	O	O

This	NN	O	O
endogenous	NN	O	B-protein
IFN	NN	O	I-protein
was	NN	O	O
also	NN	O	O
shown	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
maintaining	NN	O	O
the	NN	O	O
basal	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
class	NN	O	I-DNA
I	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
IFN-alpha5	NN	O	B-protein
and	NN	O	O
IFN-beta	NN	O	B-protein
are	NN	O	O
produced	NN	O	O
at	NN	O	O
low	NN	O	O
levels	NN	O	O
in	NN	O	O
normal	NN	O	O
tissues	NN	O	O
and	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cell	NN	O	O
function	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
maintenance	NN	O	O
of	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
IL-2	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-2	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
chain	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
composite	NN	O	B-DNA
element	NN	O	I-DNA
that	NN	O	O
binds	NN	O	O
Stat5	NN	O	B-protein
,	NN	O	O
Elf-1	NN	O	B-protein
,	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
a	NN	O	O
GATA	NN	O	B-protein
family	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin-2	NN	O	I-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
potently	NN	O	O
upregulated	NN	O	O
by	NN	O	O
its	NN	O	O
own	NN	O	O
ligand	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
characterize	NN	O	O
an	NN	O	O
essential	NN	O	O
upstream	NN	O	O
IL-2	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
that	NN	O	O
contains	NN	O	O
both	NN	O	O
consensus	NN	O	B-DNA
and	NN	O	I-DNA
non-consensus	NN	O	I-DNA
GAS	NN	O	I-DNA
motifs	NN	O	I-DNA
,	NN	O	O
two	NN	O	O
putative	NN	O	O
Ets	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
(	NN	O	O
EBS	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
overlaps	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
GAS	NN	O	I-DNA
motif	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
GATA	NN	O	B-DNA
motif	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
overlaps	NN	O	O
the	NN	O	O
non-consensus	NN	O	B-DNA
GAS	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
although	NN	O	O
the	NN	O	O
individual	NN	O	O
components	NN	O	O
of	NN	O	O
this	NN	O	O
element	NN	O	O
do	NN	O	O
not	NN	O	O
respond	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
together	NN	O	O
they	NN	O	O
form	NN	O	O
a	NN	O	O
composite	NN	O	O
element	NN	O	O
capable	NN	O	O
of	NN	O	O
conferring	NN	O	O
IL-2	NN	O	B-protein
responsiveness	NN	O	O
to	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Multiple	NN	O	O
factors	NN	O	O
including	NN	O	O
Stat5	NN	O	B-protein
,	NN	O	O
Elf-1	NN	O	B-protein
,	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
GATA	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
bind	NN	O	O
to	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
and	NN	O	O
mutation	NN	O	O
of	NN	O	O
any	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
binding	NN	O	O
sites	NN	O	O
diminishes	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
element	NN	O	O
.	NN	O	O

An	NN	O	O
unidentified	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
protein	NN	O	I-protein
binds	NN	O	O
to	NN	O	O
the	NN	O	O
EBS	NN	O	B-DNA
overlapping	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
GAS	NN	O	I-DNA
motif	NN	O	I-DNA
and	NN	O	O
appears	NN	O	O
to	NN	O	O
negatively	NN	O	O
regulate	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
requires	NN	O	O
a	NN	O	O
complex	NN	O	B-DNA
upstream	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
appears	NN	O	O
to	NN	O	O
contain	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
both	NN	O	O
positive	NN	O	B-protein
and	NN	O	I-protein
negative	NN	O	I-protein
regulatory	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Lymphocytes	NN	O	B-cell_type
from	NN	O	O
CML	NN	O	O
patients	NN	O	O
lack	NN	O	O
a	NN	O	O
47	NN	O	B-protein
kDa	NN	O	I-protein
factor	NN	O	I-protein
having	NN	O	O
affinity	NN	O	O
for	NN	O	O
a	NN	O	O
genomic	NN	O	B-DNA
sterol	NN	O	I-DNA
regulatory	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Deranged	NN	O	O
cellular	NN	O	O
cholesterol	NN	O	O
homeostasis	NN	O	O
has	NN	O	O
been	NN	O	O
widely	NN	O	O
recognized	NN	O	O
in	NN	O	O
the	NN	O	O
initiation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
progression	NN	O	O
of	NN	O	O
various	NN	O	O
types	NN	O	O
of	NN	O	O
cancers	NN	O	O
including	NN	O	O
chronic	NN	O	O
myeloid	NN	O	O
leukaemia	NN	O	O
(	NN	O	O
CML	NN	O	O
)	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
genomic	NN	O	I-DNA
sterol	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
SRE	NN	O	B-DNA
)	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
regulate	NN	O	O
various	NN	O	O
key	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
,	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
revealed	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
unique	NN	O	O
47	NN	O	B-protein
kDa	NN	O	I-protein
protein	NN	O	I-protein
factor	NN	O	I-protein
having	NN	O	O
affinity	NN	O	O
for	NN	O	O
this	NN	O	O
SRE	NN	O	B-DNA
sequence	NN	O	I-DNA
in	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
normal	NN	O	O
subjects	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
its	NN	O	O
absence	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
untreated	NN	O	O
CML	NN	O	O
patients	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
factor	NN	O	O
appeared	NN	O	O
when	NN	O	O
these	NN	O	O
CML	NN	O	O
patients	NN	O	O
achieved	NN	O	O
complete	NN	O	O
haematological	NN	O	O
remission	NN	O	O
(	NN	O	O
CHR	NN	O	O
)	NN	O	O
through	NN	O	O
alpha-interferon	NN	O	B-protein
therapy	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
an	NN	O	O
inverse	NN	O	O
relationship	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
between	NN	O	O
the	NN	O	O
LDL	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
and	NN	O	O
the	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
this	NN	O	O
47	NN	O	B-protein
kDa	NN	O	I-protein
protein	NN	O	I-protein
factor	NN	O	I-protein
to	NN	O	O
the	NN	O	O
SRE	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Based	NN	O	O
upon	NN	O	O
these	NN	O	O
results	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
this	NN	O	O
factor	NN	O	O
may	NN	O	O
have	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
chronic	NN	O	O
myeloid	NN	O	O
leukaemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
beta	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
human	NN	O	B-protein
proximal	NN	O	I-protein
sequence	NN	O	I-protein
element-binding	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
its	NN	O	O
involvement	NN	O	O
in	NN	O	O
transcription	NN	O	O
of	NN	O	O
small	NN	O	B-DNA
nuclear	NN	O	I-DNA
RNA	NN	O	I-DNA
genes	NN	O	I-DNA
by	NN	O	O
RNA	NN	O	B-protein
polymerases	NN	O	I-protein
II	NN	O	I-protein
and	NN	O	I-protein
III	NN	O	I-protein
.	NN	O	O

The	NN	O	O
proximal	NN	O	B-protein
sequence	NN	O	I-protein
element	NN	O	I-protein
(	NN	O	I-protein
PSE	NN	O	I-protein
)	NN	O	I-protein
-binding	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
PTF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
binds	NN	O	O
the	NN	O	O
PSE	NN	O	B-DNA
of	NN	O	O
both	NN	O	O
RNA	NN	O	B-DNA
polymerase	NN	O	I-DNA
II	NN	O	I-DNA
-and	NN	O	I-DNA
RNA	NN	O	I-DNA
polymerase	NN	O	I-DNA
III-transcribed	NN	O	I-DNA
mammalian	NN	O	I-DNA
small	NN	O	I-DNA
nuclear	NN	O	I-DNA
RNA	NN	O	I-DNA
(	NN	O	I-DNA
snRNA	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
their	NN	O	O
transcription	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
the	NN	O	O
purification	NN	O	O
of	NN	O	O
human	NN	O	B-protein
PTF	NN	O	I-protein
,	NN	O	O
a	NN	O	O
complex	NN	O	O
of	NN	O	O
four	NN	O	B-protein
subunits	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
molecular	NN	O	O
cloning	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
PTF	NN	O	B-protein
gamma	NN	O	I-protein
and	NN	O	I-protein
delta	NN	O	I-protein
subunits	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
isolation	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
encoding	NN	O	O
PTF	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
functional	NN	O	O
studies	NN	O	O
using	NN	O	O
anti-PTF	NN	O	B-protein
beta	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

Native	NN	O	O
PTF	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
in	NN	O	O
either	NN	O	O
protein	NN	O	O
fractions	NN	O	O
or	NN	O	O
a	NN	O	O
PTF-Oct-1-DNA	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
can	NN	O	O
be	NN	O	O
recognized	NN	O	O
by	NN	O	O
polyclonal	NN	O	B-protein
antibodies	NN	O	I-protein
raised	NN	O	O
against	NN	O	O
recombinant	NN	O	B-protein
PTF	NN	O	I-protein
beta	NN	O	I-protein
.	NN	O	O

Immunodepletion	NN	O	O
studies	NN	O	O
show	NN	O	O
that	NN	O	O
PTF	NN	O	B-protein
beta	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
transcription	NN	O	O
of	NN	O	O
both	NN	O	O
classes	NN	O	O
of	NN	O	O
snRNA	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
immunoprecipitation	NN	O	O
analyses	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
substantial	NN	O	O
and	NN	O	O
similar	NN	O	O
molar	NN	O	O
amounts	NN	O	O
of	NN	O	O
TATA-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
TBP	NN	O	B-protein
)	NN	O	O
and	NN	O	O
TFIIIB90	NN	O	B-protein
can	NN	O	O
weakly	NN	O	O
associate	NN	O	O
with	NN	O	O
PTF	NN	O	B-protein
at	NN	O	O
low	NN	O	O
salt	NN	O	O
conditions	NN	O	O
,	NN	O	O
but	NN	O	O
this	NN	O	O
association	NN	O	O
is	NN	O	O
dramatically	NN	O	O
reduced	NN	O	O
at	NN	O	O
high	NN	O	O
salt	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Along	NN	O	O
with	NN	O	O
our	NN	O	O
previous	NN	O	O
demonstration	NN	O	O
of	NN	O	O
both	NN	O	O
physical	NN	O	O
interactions	NN	O	O
between	NN	O	O
PTF	NN	O	B-protein
gamma/PTF	NN	O	I-protein
delta	NN	O	I-protein
and	NN	O	O
TBP	NN	O	B-protein
and	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
TFIIIB90	NN	O	B-protein
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
III	NN	O	I-DNA
snRNA	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
these	NN	O	O
results	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
a	NN	O	O
TBP-containing	NN	O	B-protein
complex	NN	O	I-protein
related	NN	O	O
to	NN	O	O
TFIIIB	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
III	NN	O	I-DNA
snRNA	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
acts	NN	O	O
through	NN	O	O
weak	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
four-subunit	NN	O	B-protein
PTF	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Attenuated	NN	O	O
function	NN	O	O
of	NN	O	O
a	NN	O	O
variant	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
helix-loop-helix	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
Id-3	NN	O	B-protein
,	NN	O	O
generated	NN	O	O
by	NN	O	O
an	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
mechanism	NN	O	O
.	NN	O	O

The	NN	O	O
Id	NN	O	O
family	NN	O	O
of	NN	O	O
helix-loop-helix	NN	O	B-protein
proteins	NN	O	I-protein
function	NN	O	O
as	NN	O	O
negative	NN	O	O
regulators	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
basic	NN	O	O
helix-loop-helix	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
on	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
17	NN	O	B-protein
kDa	NN	O	I-protein
variant	NN	O	I-protein
of	NN	O	O
the	NN	O	O
14	NN	O	B-protein
kDa	NN	O	I-protein
Id-3	NN	O	I-protein
protein	NN	O	O
termed	NN	O	O
Id-3L	NN	O	B-protein
(	NN	O	O
long	NN	O	O
version	NN	O	O
)	NN	O	O
which	NN	O	O
possesses	NN	O	O
a	NN	O	O
unique	NN	O	O
60	NN	O	B-protein
amino	NN	O	I-protein
acid	NN	O	I-protein
carboxy-terminus	NN	O	I-protein
generated	NN	O	O
by	NN	O	O
read	NN	O	O
through	NN	O	O
of	NN	O	O
a	NN	O	O
'	NN	O	O
coding	NN	O	B-DNA
intron	NN	O	I-DNA
'	NN	O	O
and	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
.	NN	O	O

Northern	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
minor	NN	O	O
1.1	NN	O	B-RNA
kb	NN	O	I-RNA
Id-3L	NN	O	I-RNA
transcript	NN	O	I-RNA
together	NN	O	O
with	NN	O	O
the	NN	O	O
predominant	NN	O	O
0.95	NN	O	B-RNA
kb	NN	O	I-RNA
Id-3	NN	O	I-RNA
transcript	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
adult	NN	O	O
human	NN	O	O
tissues	NN	O	O
analysed	NN	O	O
.	NN	O	O

The	NN	O	O
variant	NN	O	O
Id-3L	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
functionally	NN	O	O
distinguishable	NN	O	O
from	NN	O	O
conventional	NN	O	O
Id-3	NN	O	B-protein
since	NN	O	O
in	NN	O	O
in	NN	O	O
vitro	NN	O	O
DNA	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
greatly	NN	O	O
impaired	NN	O	O
in	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
abrogate	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
basic	NN	O	O
helix-loop-helix	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
E47	NN	O	B-protein
,	NN	O	O
to	NN	O	O
an	NN	O	O
E	NN	O	B-DNA
box	NN	O	I-DNA
recognition	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Tyloxapol	NN	O	O
inhibits	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
cytokine	NN	O	B-protein
release	NN	O	O
,	NN	O	O
scavenges	NN	O	O
HOCI	NN	O	B-protein
,	NN	O	O
and	NN	O	O
reduces	NN	O	O
viscosity	NN	O	O
of	NN	O	O
cystic	NN	O	O
fibrosis	NN	O	O
sputum	NN	O	O
.	NN	O	O

Cystic	NN	O	O
fibrosis	NN	O	O
(	NN	O	O
CF	NN	O	O
)	NN	O	O
patients	NN	O	O
develop	NN	O	O
progressive	NN	O	O
cytokine	NN	O	B-protein
-mediated	NN	O	O
inflammatory	NN	O	O
lung	NN	O	O
disease	NN	O	O
,	NN	O	O
with	NN	O	O
abundant	NN	O	O
production	NN	O	O
of	NN	O	O
thick	NN	O	O
,	NN	O	O
tenacious	NN	O	O
,	NN	O	O
protease	NN	O	B-protein
-and	NN	O	O
oxidant-rich	NN	O	O
purulent	NN	O	O
airway	NN	O	O
secretions	NN	O	O
that	NN	O	O
are	NN	O	O
difficult	NN	O	O
to	NN	O	O
clear	NN	O	O
even	NN	O	O
with	NN	O	O
physiotherapy	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
search	NN	O	O
for	NN	O	O
a	NN	O	O
potential	NN	O	O
treatment	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
tested	NN	O	O
tyloxapol	NN	O	O
,	NN	O	O
an	NN	O	O
alkylaryl	NN	O	O
polyether	NN	O	O
alcohol	NN	O	O
polymer	NN	O	O
detergent	NN	O	O
previously	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
mucolytic	NN	O	O
agent	NN	O	O
in	NN	O	O
adult	NN	O	O
chronic	NN	O	O
bronchitis	NN	O	O
.	NN	O	O

Tyloxapol	NN	O	O
inhibits	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NK-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
reduces	NN	O	O
resting	NN	O	O
secretion	NN	O	O
of	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
interleukin-8	NN	O	B-protein
(	NN	O	O
IL-8	NN	O	B-protein
)	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
monocytes	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
inhibits	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-stimulated	NN	O	O
release	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
eiconsanoids	NN	O	O
thromboxane	NN	O	O
A2	NN	O	O
and	NN	O	O
leukotriene	NN	O	O
B4	NN	O	O
(	NN	O	O
LTB4	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
tyloxapol	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
antioxidant	NN	O	O
for	NN	O	O
hydroxyl	NN	O	O
radicals	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
.	NN	O	O

Tyloxapol	NN	O	O
(	NN	O	O
0.05	NN	O	O
to	NN	O	O
0.1	NN	O	O
%	NN	O	O
wt/vol	NN	O	O
)	NN	O	O
effectively	NN	O	O
scavenges	NN	O	O
the	NN	O	O
oxidant	NN	O	O
hypochlorous	NN	O	O
acid	NN	O	O
(	NN	O	O
HOCl	NN	O	O
;	NN	O	O
1	NN	O	O
to	NN	O	O
7.5	NN	O	O
mM	NN	O	O
)	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
and	NN	O	O
protects	NN	O	O
from	NN	O	O
HOCl-mediated	NN	O	O
lung	NN	O	O
injury	NN	O	O
in	NN	O	O
rats	NN	O	O
.	NN	O	O

Tyloxapol	NN	O	O
also	NN	O	O
reduces	NN	O	O
the	NN	O	O
viscosity	NN	O	O
of	NN	O	O
CF	NN	O	O
sputum	NN	O	O
(	NN	O	O
from	NN	O	O
463	NN	O	O
+/-	NN	O	O
133	NN	O	O
to	NN	O	O
128	NN	O	O
+/-	NN	O	O
52	NN	O	O
centipoise	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
tyloxapol	NN	O	O
is	NN	O	O
potentially	NN	O	O
useful	NN	O	O
as	NN	O	O
a	NN	O	O
new	NN	O	O
antiinflammatory	NN	O	O
therapy	NN	O	O
for	NN	O	O
CF	NN	O	O
lung	NN	O	O
disease	NN	O	O
,	NN	O	O
and	NN	O	O
could	NN	O	O
possibly	NN	O	O
promote	NN	O	O
clearance	NN	O	O
of	NN	O	O
secretions	NN	O	O
in	NN	O	O
the	NN	O	O
CF	NN	O	O
airway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Elevated	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
inhibits	NN	O	O
NF-kappaB	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
in	NN	O	O
human	NN	O	O
monocytic	NN	O	O
cells	NN	O	O
and	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
regulates	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
genes	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
agents	NN	O	O
that	NN	O	O
elevate	NN	O	O
intracellular	NN	O	O
cAMP	NN	O	O
inhibit	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNFalpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
endothelial	NN	O	B-protein
leukocyte	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
vascular	NN	O	B-DNA
cell	NN	O	I-DNA
adhesion	NN	O	I-DNA
molecule-1	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
forskolin	NN	O	O
and	NN	O	O
dibutyryl	NN	O	O
cAMP	NN	O	O
,	NN	O	O
which	NN	O	O
elevate	NN	O	O
intracellular	NN	O	O
cAMP	NN	O	O
by	NN	O	O
independent	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
inhibited	NN	O	O
TNFalpha	NN	O	B-protein
and	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
expression	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
elevated	NN	O	O
cAMP	NN	O	O
and	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	O
PKA	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Elevated	NN	O	O
cAMP	NN	O	O
did	NN	O	O
not	NN	O	O
prevent	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
p50/p65	NN	O	B-protein
and	NN	O	O
c-Rel/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
,	NN	O	O
decrease	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
or	NN	O	O
significantly	NN	O	O
modify	NN	O	O
TNFalpha	NN	O	B-protein
-induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

Functional	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
plasmid	NN	O	B-DNA
containing	NN	O	O
multimerized	NN	O	B-DNA
kappaB	NN	O	I-DNA
sites	NN	O	I-DNA
by	NN	O	O
p65	NN	O	B-protein
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
agents	NN	O	O
that	NN	O	O
elevate	NN	O	O
cAMP	NN	O	O
and	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
PKA	NN	O	B-protein
.	NN	O	O

This	NN	O	O
study	NN	O	O
indicates	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
PKA	NN	O	B-protein
reduces	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
distinct	NN	O	O
set	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
inhibiting	NN	O	O
NF-kappaB	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
steroid	NN	O	O
action	NN	O	O
:	NN	O	O
a	NN	O	O
novel	NN	O	O
type	NN	O	O
of	NN	O	O
cross-talk	NN	O	O
between	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Despite	NN	O	O
the	NN	O	O
widespread	NN	O	O
use	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
diseases	NN	O	O
characterized	NN	O	O
by	NN	O	O
inflammation	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
by	NN	O	O
which	NN	O	O
these	NN	O	O
hormones	NN	O	O
exert	NN	O	O
this	NN	O	O
beneficial	NN	O	O
effect	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
asthma	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
as	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
are	NN	O	O
likely	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
causal	NN	O	O
role	NN	O	O
in	NN	O	O
inflammation	NN	O	O
in	NN	O	O
promoting	NN	O	O
cell-cell	NN	O	O
and	NN	O	O
cell-matrix	NN	O	O
interactions	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
in	NN	O	O
a	NN	O	O
monocytic	NN	O	B-cell_line
(	NN	O	I-cell_line
U937	NN	O	I-cell_line
)	NN	O	I-cell_line
and	NN	O	O
a	NN	O	O
bronchial	NN	O	B-cell_line
epithelial	NN	O	I-cell_line
(	NN	O	I-cell_line
H292	NN	O	I-cell_line
)	NN	O	I-cell_line
cell-line	NN	O	I-cell_line
dexamethasone	NN	O	O
strongly	NN	O	O
suppressed	NN	O	O
basal	NN	O	O
and	NN	O	O
induced	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Subsequent	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
ICAM-1	NN	O	I-DNA
promoter	NN	O	I-DNA
has	NN	O	O
revealed	NN	O	O
that	NN	O	O
both	NN	O	O
12-O-tetradecanoylphorbol	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
and	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
upregulate	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
through	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
)	NN	O	I-DNA
target	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
TGGAAATTCC	NN	O	O
)	NN	O	O
.	NN	O	O

No	NN	O	O
glucocorticoid	NN	O	B-DNA
recognition	NN	O	I-DNA
sequences	NN	O	I-DNA
are	NN	O	O
present	NN	O	O
in	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
and	NN	O	O
dexamethasone	NN	O	O
is	NN	O	O
still	NN	O	O
able	NN	O	O
to	NN	O	O
repress	NN	O	O
transcription	NN	O	O
when	NN	O	O
the	NN	O	O
multimerized	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
sequence	NN	O	I-DNA
is	NN	O	O
transactivated	NN	O	O
by	NN	O	O
TNF-alpha	NN	O	B-protein
upon	NN	O	O
transfection	NN	O	O
in	NN	O	O
293	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
direct	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
factors	NN	O	I-protein
is	NN	O	O
at	NN	O	O
least	NN	O	O
a	NN	O	O
partial	NN	O	O
explanation	NN	O	O
for	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
this	NN	O	O
hormone	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

Abnormalities	NN	O	O
of	NN	O	O
p16	NN	O	B-DNA
,	NN	O	I-DNA
p15	NN	O	I-DNA
and	NN	O	I-DNA
CDK4	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
recurrent	NN	O	O
malignant	NN	O	O
astrocytomas	NN	O	O
.	NN	O	O

Abnormalities	NN	O	O
in	NN	O	O
the	NN	O	O
p16	NN	O	B-DNA
,	NN	O	I-DNA
p15	NN	O	I-DNA
and	NN	O	I-DNA
CDK4	NN	O	I-DNA
genes	NN	O	I-DNA
that	NN	O	O
regulate	NN	O	O
transition	NN	O	O
through	NN	O	O
the	NN	O	O
G1	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
malignant	NN	O	O
progression	NN	O	O
of	NN	O	O
astrocytomas	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
dysfunction	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	B-DNA
also	NN	O	O
occurs	NN	O	O
during	NN	O	O
recurrence	NN	O	O
of	NN	O	O
glial	NN	O	O
tumors	NN	O	O
that	NN	O	O
were	NN	O	O
highly	NN	O	O
malignant	NN	O	O
at	NN	O	O
first	NN	O	O
presentation	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
10	NN	O	O
matched	NN	O	O
pairs	NN	O	O
of	NN	O	O
high	NN	O	O
grade	NN	O	O
malignant	NN	O	O
astrocytomas	NN	O	O
and	NN	O	O
their	NN	O	O
subsequent	NN	O	O
recurrences	NN	O	O
identified	NN	O	O
three	NN	O	O
distinct	NN	O	O
groups	NN	O	O
.	NN	O	O

The	NN	O	O
primary	NN	O	O
and	NN	O	O
recurrent	NN	O	O
tumors	NN	O	O
in	NN	O	O
Group	NN	O	O
A	NN	O	O
did	NN	O	O
not	NN	O	O
show	NN	O	O
structural	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
p16	NN	O	B-DNA
,	NN	O	I-DNA
p15	NN	O	I-DNA
or	NN	O	I-DNA
CDK4	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
whereas	NN	O	O
homozygous	NN	O	O
codeletion	NN	O	O
of	NN	O	O
p16	NN	O	B-DNA
and	NN	O	O
p15	NN	O	B-DNA
was	NN	O	O
observed	NN	O	O
in	NN	O	O
both	NN	O	O
primary	NN	O	O
and	NN	O	O
recurrent	NN	O	O
tumors	NN	O	O
in	NN	O	O
Group	NN	O	O
B	NN	O	O
.	NN	O	O

The	NN	O	O
primary	NN	O	O
tumors	NN	O	O
in	NN	O	O
Group	NN	O	O
C	NN	O	O
had	NN	O	O
a	NN	O	O
normal	NN	O	O
profile	NN	O	O
of	NN	O	O
p16	NN	O	B-protein
,	NN	O	O
p15	NN	O	B-protein
and	NN	O	O
CDK4	NN	O	B-protein
at	NN	O	O
presentation	NN	O	O
.	NN	O	O

Upon	NN	O	O
recurrence	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
tumors	NN	O	O
sustained	NN	O	O
either	NN	O	O
deletion	NN	O	O
of	NN	O	O
p16	NN	O	B-protein
alone	NN	O	O
or	NN	O	O
codeletion	NN	O	O
of	NN	O	O
both	NN	O	O
p16	NN	O	B-protein
and	NN	O	O
p15	NN	O	B-protein
or	NN	O	O
amplification	NN	O	O
of	NN	O	O
CDK4	NN	O	B-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
differences	NN	O	O
between	NN	O	O
primary	NN	O	O
anaplastic	NN	O	O
astrocytomas/glioblastomas	NN	O	O
and	NN	O	O
their	NN	O	O
subsequent	NN	O	O
recurrences	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
clinically	NN	O	O
indistinguishable	NN	O	O
,	NN	O	O
may	NN	O	O
provide	NN	O	O
better	NN	O	O
therapeutic	NN	O	O
options	NN	O	O
for	NN	O	O
treatment	NN	O	O
.	NN	O	O

-DOCSTART-	O

BCL-6	NN	O	B-protein
,	NN	O	O
a	NN	O	O
POZ/zinc-finger	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
is	NN	O	O
a	NN	O	O
sequence-specific	NN	O	B-protein
transcriptional	NN	O	I-protein
repressor	NN	O	I-protein
.	NN	O	O

Approximately	NN	O	O
40	NN	O	O
%	NN	O	O
of	NN	O	O
diffuse	NN	O	O
large	NN	O	O
cell	NN	O	O
lymphoma	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
that	NN	O	O
deregulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
BCL6	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
juxtaposing	NN	O	O
heterologous	NN	O	B-DNA
promoters	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-DNA
coding	NN	O	I-DNA
domain	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
BCL6	NN	O	B-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
95-kDa	NN	O	B-protein
protein	NN	O	I-protein
containing	NN	O	O
six	NN	O	O
C-terminal	NN	O	B-protein
zinc-finger	NN	O	I-protein
motifs	NN	O	I-protein
and	NN	O	O
an	NN	O	O
N-terminal	NN	O	B-protein
POZ	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
it	NN	O	O
may	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

By	NN	O	O
using	NN	O	O
a	NN	O	O
DNA	NN	O	B-DNA
sequence	NN	O	I-DNA
selected	NN	O	O
for	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
bind	NN	O	O
recombinant	NN	O	B-protein
BCL-6	NN	O	I-protein
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
BCL-6	NN	O	B-protein
is	NN	O	O
present	NN	O	O
in	NN	O	O
DNA-binding	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
various	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
transient	NN	O	O
transfectin	NN	O	O
experiments	NN	O	O
,	NN	O	O
BCL6	NN	O	B-protein
can	NN	O	O
repress	NN	O	O
transcription	NN	O	O
from	NN	O	O
promoters	NN	O	B-DNA
linked	NN	O	O
to	NN	O	O
its	NN	O	O
DNA	NN	O	B-DNA
target	NN	O	I-DNA
sequence	NN	O	I-DNA
and	NN	O	O
this	NN	O	O
activity	NN	O	O
is	NN	O	O
dependent	NN	O	O
upon	NN	O	O
specific	NN	O	O
DNA-binding	NN	O	O
and	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
intact	NN	O	O
N-terminal	NN	O	B-protein
half	NN	O	I-protein
of	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
this	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
BCL6	NN	O	B-protein
molecule	NN	O	I-protein
contains	NN	O	O
an	NN	O	O
autonomous	NN	O	B-protein
transrepressor	NN	O	I-protein
domain	NN	O	I-protein
and	NN	O	O
that	NN	O	O
two	NN	O	O
noncontiguous	NN	O	B-protein
regions	NN	O	I-protein
,	NN	O	O
including	NN	O	O
the	NN	O	O
POZ	NN	O	B-protein
motif	NN	O	I-protein
,	NN	O	O
mediate	NN	O	O
maximum	NN	O	O
transrepressive	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
protein	NN	O	I-protein
can	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
sequence-specific	NN	O	B-protein
transcriptional	NN	O	I-protein
repressor	NN	O	I-protein
and	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
BCL6	NN	O	B-protein
in	NN	O	O
normal	NN	O	O
lymphoid	NN	O	O
development	NN	O	O
and	NN	O	O
lymphomagenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signals	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
are	NN	O	O
stronger	NN	O	O
in	NN	O	O
neonatal	NN	O	B-cell_type
than	NN	O	I-cell_type
adult	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
background	NN	O	O
of	NN	O	O
the	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
reactivity	NN	O	O
of	NN	O	O
human	NN	O	O
neonates	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
fully	NN	O	O
elucidated	NN	O	O
.	NN	O	O

As	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
has	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
several	NN	O	O
genes	NN	O	B-DNA
involved	NN	O	O
in	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
,	NN	O	O
the	NN	O	O
authors	NN	O	O
have	NN	O	O
analysed	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
cord	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
activity	NN	O	O
was	NN	O	O
tested	NN	O	O
by	NN	O	O
quantitating	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
an	NN	O	O
oligonucleotide	NN	O	O
containing	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
obtained	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
phorbol	NN	O	O
dibutyrate/calcium	NN	O	O
ionophore	NN	O	O
A23187	NN	O	O
(	NN	O	O
PDBu/iono	NN	O	O
)	NN	O	O
combination	NN	O	O
induced	NN	O	O
a	NN	O	O
clearly	NN	O	O
higher	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
neonatal	NN	O	B-cell_type
than	NN	O	I-cell_type
adult	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
higher	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
the	NN	O	O
CD4+	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
subset	NN	O	I-cell_type
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
with	NN	O	O
antibodies	NN	O	B-protein
directed	NN	O	O
against	NN	O	O
the	NN	O	O
major	NN	O	O
components	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
the	NN	O	O
p50	NN	O	B-protein
and	NN	O	I-protein
RelA	NN	O	I-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
composition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
similar	NN	O	O
in	NN	O	O
neonatal	NN	O	B-cell_type
and	NN	O	I-cell_type
adult	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
exposed	NN	O	O
to	NN	O	O
oxidative	NN	O	O
stress-inducing	NN	O	O
signals	NN	O	O
during	NN	O	O
delivery	NN	O	O
and/or	NN	O	O
are	NN	O	O
inherently	NN	O	O
more	NN	O	O
sensitive	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activating	NN	O	O
signals	NN	O	O
than	NN	O	O
adult	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Dual	NN	O	O
action	NN	O	O
of	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
on	NN	O	O
human	NN	O	O
embryonic/fetal	NN	O	O
hematopoiesis	NN	O	O
:	NN	O	O
blockade	NN	O	O
of	NN	O	O
primitive	NN	O	O
progenitor	NN	O	O
proliferation	NN	O	O
and	NN	O	O
shift	NN	O	O
from	NN	O	O
multipotent/erythroid/monocytic	NN	O	O
to	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
program	NN	O	O
.	NN	O	O

In	NN	O	O
preliminary	NN	O	O
studies	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
HGF	NN	O	B-protein
)	NN	O	O
requirement	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HPCs	NN	O	B-cell_type
)	NN	O	O
purified	NN	O	O
from	NN	O	O
embryonic-fetal	NN	O	O
liver	NN	O	O
(	NN	O	O
FL	NN	O	O
)	NN	O	O
and	NN	O	O
grown	NN	O	O
in	NN	O	O
fetal	NN	O	O
calf	NN	O	O
serum-supplemented	NN	O	O
(	NN	O	O
FCS+	NN	O	O
)	NN	O	O
clonogenic	NN	O	O
culture	NN	O	O
.	NN	O	O

The	NN	O	O
key	NN	O	O
role	NN	O	O
of	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
Epo	NN	O	B-protein
)	NN	O	O
for	NN	O	O
colony	NN	O	O
formation	NN	O	O
by	NN	O	O
early	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
(	NN	O	O
burst-forming	NN	O	B-cell_type
units-erythroid	NN	O	I-cell_type
[	NN	O	O
BFU-E	NN	O	B-cell_type
]	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
confirmed	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
exogenous	NN	O	O
HGFs	NN	O	B-protein
,	NN	O	O
FL	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
(	NN	O	O
colony-forming	NN	O	B-cell_type
unit	NN	O	I-cell_type
monocyte	NN	O	I-cell_type
[	NN	O	O
CFU-M	NN	O	B-cell_type
]	NN	O	O
)	NN	O	O
generate	NN	O	O
large	NN	O	O
colonies	NN	O	O
exclusively	NN	O	O
composed	NN	O	O
of	NN	O	O
monocytes-macrophages	NN	O	O
;	NN	O	O
these	NN	O	O
colonies	NN	O	O
are	NN	O	O
absent	NN	O	O
in	NN	O	O
FCS-	NN	O	O
clonogenic	NN	O	O
culture	NN	O	O
.	NN	O	O

On	NN	O	O
this	NN	O	O
basis	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
and	NN	O	O
its	NN	O	O
isomer	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
in	NN	O	O
FL	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

Both	NN	O	O
compounds	NN	O	O
modulate	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
purified	NN	O	B-cell_line
FL	NN	O	I-cell_line
HPCs	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
show	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
shift	NN	O	O
from	NN	O	O
mixed/erythroid/monocytic	NN	O	O
to	NN	O	O
granulocytic	NN	O	O
colony	NN	O	O
formation	NN	O	O
.	NN	O	O

Studies	NN	O	O
on	NN	O	O
unicellular	NN	O	B-cell_line
and	NN	O	I-cell_line
paired	NN	O	I-cell_line
daughter	NN	O	I-cell_line
cell	NN	O	I-cell_line
culture	NN	O	I-cell_line
unequivocally	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
shift	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
HPC	NN	O	O
differentiation	NN	O	O
program	NN	O	O
to	NN	O	O
the	NN	O	O
granulopoietic	NN	O	O
pathway	NN	O	O
(	NN	O	O
rather	NN	O	O
than	NN	O	O
RA-induced	NN	O	O
down-modulation	NN	O	O
of	NN	O	O
multipotent	NN	O	O
/erythroid/monocytic	NN	O	O
HPC	NN	O	O
growth	NN	O	O
coupled	NN	O	O
with	NN	O	O
recruitment	NN	O	O
of	NN	O	O
granulocytic	NN	O	B-cell_type
HPCs	NN	O	I-cell_type
)	NN	O	O
.	NN	O	O

ATRA	NN	O	O
and	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
also	NN	O	O
exert	NN	O	O
their	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
primitive	NN	O	B-cell_type
HPCs	NN	O	I-cell_type
(	NN	O	O
high-proliferative	NN	O	B-cell_line
potential	NN	O	I-cell_line
colony-forming	NN	O	I-cell_line
cells	NN	O	I-cell_line
[	NN	O	O
HPP-CFCs	NN	O	B-cell_line
]	NN	O	O
)	NN	O	O
and	NN	O	O
putative	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HSCs	NN	O	B-cell_type
;	NN	O	O
assayed	NN	O	O
in	NN	O	O
Dexter-type	NN	O	O
long-term	NN	O	O
culture	NN	O	O
)	NN	O	O
.	NN	O	O

High	NN	O	O
concentrations	NN	O	O
of	NN	O	O
either	NN	O	O
compound	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
drastically	NN	O	O
reduced	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
primary	NN	O	O
HPP-CFC	NN	O	B-cell_line
colonies	NN	O	I-cell_line
and	NN	O	O
totally	NN	O	O
abolished	NN	O	O
their	NN	O	O
recloning	NN	O	O
capacity	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
inhibited	NN	O	O
HSC	NN	O	B-cell_type
proliferation	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
crucial	NN	O	O
that	NN	O	O
these	NN	O	O
results	NN	O	O
mirror	NN	O	O
recent	NN	O	O
observations	NN	O	O
indicating	NN	O	O
that	NN	O	O
murine	NN	O	O
adult	NN	O	O
HPCs	NN	O	B-cell_type
transduced	NN	O	O
with	NN	O	O
dominant	NN	O	O
negative	NN	O	O
ATRA	NN	O	B-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
RAR	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
are	NN	O	O
immortalized	NN	O	O
and	NN	O	O
show	NN	O	O
a	NN	O	O
selective	NN	O	O
blockade	NN	O	O
of	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
ATRA/9-cis	NN	O	O
RA	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
FL	NN	O	O
hematopoiesis	NN	O	O
via	NN	O	O
a	NN	O	O
dual	NN	O	O
effect	NN	O	O
hypothetically	NN	O	O
mediated	NN	O	O
by	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
RAR/RXR	NN	O	B-protein
heterodimer	NN	O	I-protein
,	NN	O	O
ie	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
HSC/	NN	O	B-cell_line
primitive	NN	O	I-cell_line
HPC	NN	O	I-cell_line
proliferation	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
CFU-GEMM/	NN	O	O
BFU-E/CFU-M	NN	O	O
shift	NN	O	O
from	NN	O	O
the	NN	O	O
multipotent/erythroid/monocytic	NN	O	O
to	NN	O	O
the	NN	O	O
granulocytic-neutrophilic	NN	O	O
differentiation	NN	O	O
program	NN	O	O
.	NN	O	O

-DOCSTART-	O

Dexamethasone	NN	O	O
suppression	NN	O	O
test	NN	O	O
:	NN	O	O
corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
regulation	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
of	NN	O	O
young	NN	O	O
and	NN	O	O
aged	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
dexamethasone	NN	O	O
suppression	NN	O	O
test	NN	O	O
(	NN	O	O
DST	NN	O	O
)	NN	O	O
is	NN	O	O
considered	NN	O	O
an	NN	O	O
indicator	NN	O	O
of	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
adrenal	NN	O	O
pituitary	NN	O	O
axis	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
its	NN	O	O
binding	NN	O	O
to	NN	O	O
corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
previously	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
measurement	NN	O	O
of	NN	O	O
corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
is	NN	O	O
an	NN	O	O
index	NN	O	O
of	NN	O	O
an	NN	O	O
analogous	NN	O	O
pattern	NN	O	O
in	NN	O	O
brain	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
corticosteroid	NN	O	B-protein
Type	NN	O	I-protein
I	NN	O	I-protein
and	NN	O	I-protein
Type	NN	O	I-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
were	NN	O	O
measured	NN	O	O
in	NN	O	O
10	NN	O	O
elderly	NN	O	O
subjects	NN	O	O
and	NN	O	O
in	NN	O	O
9	NN	O	O
young	NN	O	O
adults	NN	O	O
,	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
overnight	NN	O	O
DST	NN	O	O
(	NN	O	O
1	NN	O	O
mg	NN	O	O
)	NN	O	O
.	NN	O	O

Receptors	NN	O	O
were	NN	O	O
measured	NN	O	O
by	NN	O	O
radioreceptor	NN	O	O
assay	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
the	NN	O	O
subjects	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
suppress	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
Type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
Type	NN	O	I-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
before	NN	O	O
the	NN	O	O
test	NN	O	O
was	NN	O	O
lower	NN	O	O
in	NN	O	O
elderly	NN	O	O
subjects	NN	O	O
than	NN	O	O
in	NN	O	O
adults	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
control	NN	O	O
group	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
produced	NN	O	O
a	NN	O	O
significant	NN	O	O
depression	NN	O	O
of	NN	O	O
Type	NN	O	B-protein
I	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
from	NN	O	O
267	NN	O	O
+/-	NN	O	O
72	NN	O	O
to	NN	O	O
169	NN	O	O
+/-	NN	O	O
71	NN	O	O
receptors	NN	O	O
per	NN	O	O
cell	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
can	NN	O	O
be	NN	O	O
interpreted	NN	O	O
as	NN	O	O
a	NN	O	O
primary	NN	O	O
involvement	NN	O	O
of	NN	O	O
Type	NN	O	B-protein
I	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
;	NN	O	O
Type	NN	O	B-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
decreased	NN	O	O
in	NN	O	O
half	NN	O	O
the	NN	O	O
subjects	NN	O	O
(	NN	O	O
from	NN	O	O
2849	NN	O	O
+/-	NN	O	O
703	NN	O	O
to	NN	O	O
2345	NN	O	O
+/-	NN	O	O
569	NN	O	O
receptors	NN	O	O
per	NN	O	O
cell	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
elderly	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
,	NN	O	O
Type	NN	O	B-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
were	NN	O	O
also	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
(	NN	O	O
from	NN	O	O
1796	NN	O	O
+/-	NN	O	O
671	NN	O	O
to	NN	O	O
720	NN	O	O
+/-	NN	O	O
345	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
in	NN	O	O
young	NN	O	O
subjects	NN	O	O
Type	NN	O	B-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
are	NN	O	O
initially	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
and	NN	O	O
then	NN	O	O
down-regulated	NN	O	O
,	NN	O	O
while	NN	O	O
in	NN	O	O
aged	NN	O	O
subjects	NN	O	O
an	NN	O	O
up-regulation	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
achieved	NN	O	O
,	NN	O	O
as	NN	O	O
suggested	NN	O	O
by	NN	O	O
the	NN	O	O
higher	NN	O	O
values	NN	O	O
of	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
usually	NN	O	O
found	NN	O	O
in	NN	O	O
aging	NN	O	O
subjects	NN	O	O
.	NN	O	O

-DOCSTART-	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
ligand-binding	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
human	NN	O	B-protein
retinoic	NN	O	I-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
by	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
.	NN	O	O

Three	NN	O	O
subtypes	NN	O	O
of	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
termed	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
RAR	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
and	NN	O	O
RAR	NN	O	B-protein
gamma	NN	O	I-protein
,	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

They	NN	O	O
are	NN	O	O
composed	NN	O	O
of	NN	O	O
different	NN	O	O
structural	NN	O	B-protein
domains	NN	O	I-protein
,	NN	O	O
including	NN	O	O
distinct	NN	O	O
domains	NN	O	O
for	NN	O	O
DNA	NN	O	O
and	NN	O	O
ligand	NN	O	O
binding	NN	O	O
.	NN	O	O

RARs	NN	O	B-protein
specifically	NN	O	O
bind	NN	O	O
all-trans-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
,	NN	O	O
9-cis-RA	NN	O	O
,	NN	O	O
and	NN	O	O
retinoid	NN	O	O
analogs	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
functional	NN	O	O
role	NN	O	O
of	NN	O	O
cysteine	NN	O	O
and	NN	O	O
arginine	NN	O	O
residues	NN	O	O
in	NN	O	O
the	NN	O	O
ligand-binding	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
hRAR	NN	O	B-protein
alpha	NN	O	I-protein
(	NN	O	O
hRAR	NN	O	B-protein
alpha-LBD	NN	O	I-protein
,	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
154	NN	O	I-protein
to	NN	O	I-protein
462	NN	O	I-protein
)	NN	O	O
.	NN	O	O

All	NN	O	O
conserved	NN	O	O
cysteine	NN	O	O
and	NN	O	O
arginine	NN	O	O
residues	NN	O	O
in	NN	O	O
this	NN	O	O
domain	NN	O	O
were	NN	O	O
mutated	NN	O	O
by	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
mutant	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
characterized	NN	O	O
by	NN	O	O
blocking	NN	O	O
reactions	NN	O	O
,	NN	O	O
ligand-binding	NN	O	O
experiments	NN	O	O
,	NN	O	O
transactivation	NN	O	O
assays	NN	O	O
,	NN	O	O
and	NN	O	O
protease	NN	O	B-protein
mapping	NN	O	O
.	NN	O	O

Changes	NN	O	O
of	NN	O	O
any	NN	O	O
cysteine	NN	O	O
residue	NN	O	O
of	NN	O	O
the	NN	O	O
hRAR	NN	O	B-protein
alpha-LBD	NN	O	I-protein
had	NN	O	O
no	NN	O	O
significant	NN	O	O
influence	NN	O	O
on	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
or	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
residue	NN	O	O
C-235	NN	O	O
is	NN	O	O
specifically	NN	O	O
important	NN	O	O
in	NN	O	O
antagonist	NN	O	O
binding	NN	O	O
.	NN	O	O

With	NN	O	O
respect	NN	O	O
to	NN	O	O
arginine	NN	O	O
residues	NN	O	O
,	NN	O	O
only	NN	O	O
the	NN	O	O
two	NN	O	O
single	NN	O	O
mutations	NN	O	O
of	NN	O	O
R-276	NN	O	O
and	NN	O	O
R-394	NN	O	O
to	NN	O	O
alanine	NN	O	O
showed	NN	O	O
a	NN	O	O
dramatic	NN	O	O
decrease	NN	O	O
of	NN	O	O
agonist	NN	O	O
and	NN	O	O
antagonist	NN	O	O
binding	NN	O	O
whereas	NN	O	O
the	NN	O	O
R272A	NN	O	O
mutation	NN	O	O
showed	NN	O	O
only	NN	O	O
a	NN	O	O
slight	NN	O	O
effect	NN	O	O
.	NN	O	O

For	NN	O	O
all	NN	O	O
other	NN	O	O
arginine	NN	O	O
mutations	NN	O	O
,	NN	O	O
no	NN	O	O
differences	NN	O	O
in	NN	O	O
affinity	NN	O	O
were	NN	O	O
detectable	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
mutations	NN	O	O
R217A	NN	O	O
and	NN	O	O
R294A	NN	O	O
caused	NN	O	O
an	NN	O	O
increased	NN	O	O
binding	NN	O	O
efficiency	NN	O	O
for	NN	O	O
antagonists	NN	O	O
but	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
agonist	NN	O	O
binding	NN	O	O
.	NN	O	O

From	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
we	NN	O	O
can	NN	O	O
conclude	NN	O	O
that	NN	O	O
electrostatic	NN	O	O
interactions	NN	O	O
of	NN	O	O
retinoids	NN	O	O
with	NN	O	O
the	NN	O	O
RAR	NN	O	B-protein
alpha-LBD	NN	O	I-protein
play	NN	O	O
a	NN	O	O
significant	NN	O	O
role	NN	O	O
in	NN	O	O
ligand	NN	O	O
binding	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
antagonists	NN	O	O
show	NN	O	O
distinctly	NN	O	O
different	NN	O	O
requirements	NN	O	O
for	NN	O	O
efficient	NN	O	O
binding	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
their	NN	O	O
interference	NN	O	O
in	NN	O	O
the	NN	O	O
ligand-inducible	NN	O	O
transactivation	NN	O	O
function	NN	O	O
of	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

IL4	NN	O	B-protein
and	NN	O	I-protein
IL13	NN	O	I-protein
receptors	NN	O	I-protein
share	NN	O	O
the	NN	O	O
gamma	NN	O	B-protein
c	NN	O	I-protein
chain	NN	O	I-protein
and	NN	O	O
activate	NN	O	O
STAT6	NN	O	B-protein
,	NN	O	I-protein
STAT3	NN	O	I-protein
and	NN	O	I-protein
STAT5	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

IL13	NN	O	B-protein
induces	NN	O	O
the	NN	O	O
same	NN	O	O
biological	NN	O	O
effects	NN	O	O
as	NN	O	O
IL4	NN	O	B-protein
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
IL4R	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
both	NN	O	O
IL4R	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
IL2R	NN	O	B-protein
gamma	NN	O	I-protein
c	NN	O	I-protein
are	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
IL13R	NN	O	B-protein
and	NN	O	O
that	NN	O	O
both	NN	O	O
cytokines	NN	O	B-protein
induced	NN	O	O
STAT6	NN	O	B-protein
,	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
spite	NN	O	O
of	NN	O	O
this	NN	O	O
similar	NN	O	O
downstream	NN	O	O
signalling	NN	O	O
,	NN	O	O
IL4	NN	O	B-protein
and	NN	O	O
IL13	NN	O	B-protein
used	NN	O	O
a	NN	O	O
different	NN	O	O
set	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
kinases	NN	O	I-protein
:	NN	O	O
IL13	NN	O	B-protein
is	NN	O	O
unable	NN	O	O
to	NN	O	O
activate	NN	O	O
JAK1	NN	O	B-protein
and	NN	O	O
JAK3	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Calcineurin	NN	O	B-protein
mutants	NN	O	I-protein
render	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
resistant	NN	O	O
to	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

The	NN	O	O
immunosuppressants	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
and	NN	O	O
FK506	NN	O	O
have	NN	O	O
been	NN	O	O
widely	NN	O	O
used	NN	O	O
to	NN	O	O
prevent	NN	O	O
and	NN	O	O
treat	NN	O	O
graft	NN	O	O
rejection	NN	O	O
after	NN	O	O
human	NN	O	O
organ	NN	O	O
and	NN	O	O
tissue	NN	O	O
transplantations	NN	O	O
.	NN	O	O

CsA	NN	O	O
and	NN	O	O
FK506	NN	O	O
associate	NN	O	O
with	NN	O	O
intracellular	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
CsA	NN	O	O
with	NN	O	O
cyclophilin	NN	O	B-protein
A	NN	O	I-protein
and	NN	O	O
FK506	NN	O	O
with	NN	O	O
FKBP12	NN	O	B-protein
)	NN	O	O
to	NN	O	O
form	NN	O	O
protein/drug	NN	O	O
complexes	NN	O	O
that	NN	O	O
suppress	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
by	NN	O	O
preventing	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
antigen	NN	O	O
presentation	NN	O	O
.	NN	O	O

The	NN	O	O
common	NN	O	O
target	NN	O	O
of	NN	O	O
CsA	NN	O	O
and	NN	O	O
FK506	NN	O	O
is	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
Ca2+/calmodulin-regulated	NN	O	B-protein
,	NN	O	I-protein
serine/threonine-specific	NN	O	I-protein
protein	NN	O	I-protein
phosphatase	NN	O	I-protein
that	NN	O	O
regulates	NN	O	O
the	NN	O	O
nuclear	NN	O	O
import	NN	O	O
of	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
required	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
T	NN	O	B-DNA
cell	NN	O	I-DNA
activation	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
previous	NN	O	O
studies	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
calcineurin	NN	O	B-protein
mutations	NN	O	O
that	NN	O	O
block	NN	O	O
binding	NN	O	O
by	NN	O	O
the	NN	O	O
cyclophilin	NN	O	B-protein
A/CsA	NN	O	I-protein
or	NN	O	I-protein
FKBP12/FK506	NN	O	I-protein
complexes	NN	O	I-protein
and	NN	O	O
thereby	NN	O	O
render	NN	O	O
yeast	NN	O	O
cells	NN	O	O
resistant	NN	O	O
to	NN	O	O
the	NN	O	O
antifungal	NN	O	O
effects	NN	O	O
of	NN	O	O
CsA	NN	O	O
or	NN	O	O
FK506	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
corresponding	NN	O	O
mutations	NN	O	O
in	NN	O	O
murine	NN	O	O
calcineurin	NN	O	B-protein
render	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
signal	NN	O	O
transduction	NN	O	O
cascade	NN	O	O
CsA	NN	O	O
resistant	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
support	NN	O	O
the	NN	O	O
recently	NN	O	O
determined	NN	O	O
calcineurin	NN	O	B-protein
X-ray	NN	O	O
crystal	NN	O	O
structure	NN	O	O
,	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
calcineurin	NN	O	B-protein
is	NN	O	O
the	NN	O	O
only	NN	O	O
CsA-sensitive	NN	O	B-protein
component	NN	O	I-protein
limiting	NN	O	O
signaling	NN	O	O
from	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
means	NN	O	O
to	NN	O	O
render	NN	O	O
cells	NN	O	O
and	NN	O	O
tissues	NN	O	O
resistant	NN	O	O
to	NN	O	O
the	NN	O	O
toxic	NN	O	O
side	NN	O	O
effects	NN	O	O
of	NN	O	O
CsA	NN	O	O
and	NN	O	O
FK506	NN	O	O
.	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	O
viral	NN	O	O
latency	NN	O	O
is	NN	O	O
disrupted	NN	O	O
by	NN	O	O
the	NN	O	O
immediate-early	NN	O	B-protein
BRLF1	NN	O	I-protein
protein	NN	O	I-protein
through	NN	O	O
a	NN	O	O
cell-specific	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
causative	NN	O	O
agent	NN	O	O
of	NN	O	O
infectious	NN	O	O
mononucleosis	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
associated	NN	O	O
with	NN	O	O
epithelial	NN	O	O
cell	NN	O	O
malignancies	NN	O	O
(	NN	O	O
nasopharyngeal	NN	O	O
carcinoma	NN	O	O
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
B-cell	NN	O	O
malignancies	NN	O	O
.	NN	O	O

Understanding	NN	O	O
how	NN	O	O
viral	NN	O	O
latency	NN	O	O
is	NN	O	O
disrupted	NN	O	O
is	NN	O	O
a	NN	O	O
central	NN	O	O
issue	NN	O	O
in	NN	O	O
herpesvirus	NN	O	O
biology	NN	O	O
.	NN	O	O

Epithelial	NN	O	O
cells	NN	O	O
are	NN	O	O
the	NN	O	O
major	NN	O	O
site	NN	O	O
of	NN	O	O
lytic	NN	O	O
EBV	NN	O	O
replication	NN	O	O
within	NN	O	O
the	NN	O	O
human	NN	O	O
host	NN	O	O
,	NN	O	O
and	NN	O	O
viral	NN	O	O
reactivation	NN	O	O
occurs	NN	O	O
in	NN	O	O
EBV-associated	NN	O	O
nasopharyngeal	NN	O	O
carcinomas	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
known	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
viral	NN	O	B-protein
immediate-early	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
BZLF1	NN	O	B-protein
,	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
initiate	NN	O	O
the	NN	O	O
switch	NN	O	O
from	NN	O	O
latent	NN	O	O
to	NN	O	O
lytic	NN	O	O
infection	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cellular	NN	O	O
regulation	NN	O	O
of	NN	O	O
BZLF1	NN	O	B-protein
transcription	NN	O	O
is	NN	O	O
therefore	NN	O	O
thought	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
stringency	NN	O	O
of	NN	O	O
viral	NN	O	O
latency	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
,	NN	O	O
unexpectedly	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
another	NN	O	O
viral	NN	O	B-protein
immediate-early	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
BRLF1	NN	O	B-protein
,	NN	O	O
can	NN	O	O
disrupt	NN	O	O
viral	NN	O	O
latency	NN	O	O
in	NN	O	O
an	NN	O	O
epithelial	NN	O	O
cell-specific	NN	O	O
fashion	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
leading	NN	O	O
to	NN	O	O
disruption	NN	O	O
of	NN	O	O
EBV	NN	O	O
latency	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
cell-type	NN	O	O
specific	NN	O	O
.	NN	O	O

-DOCSTART-	O

Eosinophil	NN	O	B-cell_type
priming	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
:	NN	O	O
from	NN	O	O
cellular	NN	O	O
signal	NN	O	O
to	NN	O	O
in	NN	O	O
vivo	NN	O	O
modulation	NN	O	O
.	NN	O	O

Eosinophils	NN	O	B-cell_type
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
effector	NN	O	O
phase	NN	O	O
of	NN	O	O
allergic	NN	O	O
inflammation	NN	O	O
.	NN	O	O

This	NN	O	O
review	NN	O	O
will	NN	O	O
focus	NN	O	O
on	NN	O	O
the	NN	O	O
conversion	NN	O	O
of	NN	O	O
the	NN	O	O
unprimed	NN	O	O
eosinophil	NN	O	O
phenotype	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
normal	NN	O	O
individuals	NN	O	O
to	NN	O	O
the	NN	O	O
primed	NN	O	O
phenotype	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
and	NN	O	O
tissues	NN	O	O
of	NN	O	O
allergic	NN	O	O
patients	NN	O	O
,	NN	O	O
a	NN	O	O
phenomenon	NN	O	O
called	NN	O	O
priming	NN	O	O
.	NN	O	O

Recent	NN	O	O
data	NN	O	O
on	NN	O	O
the	NN	O	O
signals	NN	O	O
initiated	NN	O	O
after	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
activation	NN	O	O
on	NN	O	O
eosinophils	NN	O	B-cell_type
will	NN	O	O
be	NN	O	O
reviewed	NN	O	O
.	NN	O	O

-DOCSTART-	O

CTL	NN	O	O
recognition	NN	O	O
of	NN	O	O
an	NN	O	O
altered	NN	O	O
peptide	NN	O	O
associated	NN	O	O
with	NN	O	O
asparagine	NN	O	O
bond	NN	O	O
rearrangement	NN	O	O
.	NN	O	O

Implications	NN	O	O
for	NN	O	O
immunity	NN	O	O
and	NN	O	O
vaccine	NN	O	O
design	NN	O	O
.	NN	O	O

The	NN	O	O
extent	NN	O	O
to	NN	O	O
which	NN	O	O
peptides	NN	O	O
containing	NN	O	O
chemically	NN	O	O
and	NN	O	O
post-translationally	NN	O	O
modified	NN	O	O
amino	NN	O	B-protein
acid	NN	O	I-protein
side	NN	O	I-protein
chains	NN	O	I-protein
are	NN	O	O
recognized	NN	O	O
by	NN	O	O
primed	NN	O	B-cell_line
CTL	NN	O	I-cell_line
has	NN	O	O
not	NN	O	O
been	NN	O	O
clearly	NN	O	O
defined	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
the	NN	O	O
CTL	NN	O	O
recognition	NN	O	O
of	NN	O	O
a	NN	O	O
MHC	NN	O	O
class	NN	O	O
I-restricted	NN	O	O
peptide	NN	O	O
containing	NN	O	O
a	NN	O	O
cyclized	NN	O	O
asparagine	NN	O	O
(	NN	O	O
succinimide	NN	O	O
)	NN	O	O
residue	NN	O	O
.	NN	O	O

This	NN	O	O
modification	NN	O	O
of	NN	O	O
the	NN	O	O
asparagine	NN	O	O
side	NN	O	O
chain	NN	O	O
is	NN	O	O
a	NN	O	O
common	NN	O	O
intermediate	NN	O	O
structure	NN	O	O
during	NN	O	O
deamidation	NN	O	O
,	NN	O	O
isomerization	NN	O	O
,	NN	O	O
and	NN	O	O
bond	NN	O	O
rearrangements	NN	O	O
of	NN	O	O
amide-containing	NN	O	O
amino	NN	O	O
acids	NN	O	O
and	NN	O	O
also	NN	O	O
occurs	NN	O	O
as	NN	O	O
a	NN	O	O
side	NN	O	O
reaction	NN	O	O
in	NN	O	O
peptide	NN	O	O
synthesis	NN	O	O
.	NN	O	O

The	NN	O	O
CTL	NN	O	B-cell_type
specifically	NN	O	O
recognized	NN	O	O
the	NN	O	O
succinimide-containing	NN	O	O
peptide	NN	O	O
showing	NN	O	O
only	NN	O	O
weak	NN	O	O
cross-reactivity	NN	O	O
at	NN	O	O
high	NN	O	O
concentrations	NN	O	O
of	NN	O	O
the	NN	O	O
parent	NN	O	O
peptide	NN	O	O
containing	NN	O	O
unmodified	NN	O	O
asparagine	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
CTL	NN	O	B-cell_type
raised	NN	O	O
against	NN	O	O
the	NN	O	O
parent	NN	O	O
peptide	NN	O	O
did	NN	O	O
not	NN	O	O
recognize	NN	O	O
the	NN	O	O
succinimide	NN	O	O
derivative	NN	O	O
of	NN	O	O
this	NN	O	O
peptide	NN	O	O
.	NN	O	O

Naturally	NN	O	O
processed	NN	O	O
forms	NN	O	O
of	NN	O	O
these	NN	O	O
structures	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
occur	NN	O	O
given	NN	O	O
the	NN	O	O
importance	NN	O	O
and	NN	O	O
frequency	NN	O	O
of	NN	O	O
deamidation	NN	O	O
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
since	NN	O	O
succinimide	NN	O	O
intermediates	NN	O	O
of	NN	O	O
deamidated	NN	O	O
peptides	NN	O	O
can	NN	O	O
occasionally	NN	O	O
be	NN	O	O
very	NN	O	O
stable	NN	O	O
,	NN	O	O
these	NN	O	O
peptides	NN	O	O
have	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
act	NN	O	O
as	NN	O	O
altered	NN	O	O
self-Ags	NN	O	O
with	NN	O	O
significant	NN	O	O
implications	NN	O	O
for	NN	O	O
autoimmunity	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
unwanted	NN	O	O
and	NN	O	O
potentially	NN	O	O
hazardous	NN	O	O
specificities	NN	O	O
may	NN	O	O
be	NN	O	O
elicited	NN	O	O
when	NN	O	O
using	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
in	NN	O	O
subunit	NN	O	O
vaccines	NN	O	O
in	NN	O	O
which	NN	O	O
succinimide	NN	O	O
residues	NN	O	O
may	NN	O	O
form	NN	O	O
spontaneously	NN	O	O
during	NN	O	O
storage	NN	O	O
or	NN	O	O
chemical	NN	O	O
synthesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
3	NN	O	O
'	NN	O	O
--	NN	O	O
>	NN	O	O
5	NN	O	O
'	NN	O	O
XPB	NN	O	B-protein
helicase	NN	O	I-protein
defect	NN	O	O
in	NN	O	O
repair/transcription	NN	O	B-protein
factor	NN	O	I-protein
TFIIH	NN	O	B-protein
of	NN	O	O
xeroderma	NN	O	O
pigmentosum	NN	O	O
group	NN	O	O
B	NN	O	O
affects	NN	O	O
both	NN	O	O
DNA	NN	O	O
repair	NN	O	O
and	NN	O	O
transcription	NN	O	O
.	NN	O	O

XPB	NN	O	B-protein
is	NN	O	O
a	NN	O	O
subunit	NN	O	O
of	NN	O	O
the	NN	O	O
basal	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
TFIIH	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
also	NN	O	O
involved	NN	O	O
in	NN	O	O
nucleotide	NN	O	O
excision	NN	O	O
repair	NN	O	O
(	NN	O	O
NER	NN	O	O
)	NN	O	O
and	NN	O	O
potentially	NN	O	O
in	NN	O	O
cell	NN	O	O
cycle	NN	O	O
regulation	NN	O	O
.	NN	O	O

A	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
3'-end	NN	O	O
of	NN	O	O
the	NN	O	O
XPB	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
a	NN	O	O
concurrence	NN	O	O
of	NN	O	O
two	NN	O	O
disorders	NN	O	O
:	NN	O	O
xeroderma	NN	O	O
pigmentosum	NN	O	O
(	NN	O	O
XP	NN	O	O
)	NN	O	O
and	NN	O	O
Cockayne	NN	O	O
's	NN	O	O
syndrome	NN	O	O
(	NN	O	O
CS	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
isolated	NN	O	O
TFIIH	NN	O	B-protein
from	NN	O	O
cells	NN	O	O
derived	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
(	NN	O	O
XP11BE	NN	O	O
)	NN	O	O
who	NN	O	O
carries	NN	O	O
this	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
(	NN	O	O
TFIIHmut	NN	O	B-protein
)	NN	O	O
and	NN	O	O
from	NN	O	O
the	NN	O	O
mother	NN	O	O
of	NN	O	O
this	NN	O	O
patient	NN	O	O
(	NN	O	O
TFIIHwt	NN	O	B-protein
)	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
biochemical	NN	O	O
consequences	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
.	NN	O	O

Although	NN	O	O
identical	NN	O	O
in	NN	O	O
composition	NN	O	O
and	NN	O	O
stoichiometry	NN	O	O
to	NN	O	O
TFIIHwt	NN	O	B-protein
,	NN	O	O
TFIIHmut	NN	O	B-protein
shows	NN	O	O
a	NN	O	O
reduced	NN	O	O
3	NN	O	O
'	NN	O	O
--	NN	O	O
>	NN	O	O
5	NN	O	O
'	NN	O	O
XPB	NN	O	B-protein
helicase	NN	O	O
activity	NN	O	O
.	NN	O	O

A	NN	O	O
decrease	NN	O	O
in	NN	O	O
helicase	NN	O	O
and	NN	O	O
DNA-dependent	NN	O	B-protein
ATPase	NN	O	I-protein
activities	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
with	NN	O	O
the	NN	O	O
mutated	NN	O	O
recombinant	NN	O	O
XPB	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
XPB	NN	O	B-protein
mutation	NN	O	O
causes	NN	O	O
a	NN	O	O
severe	NN	O	O
NER	NN	O	O
defect	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
basal	NN	O	O
transcription	NN	O	O
activity	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
latter	NN	O	O
defect	NN	O	O
may	NN	O	O
provide	NN	O	O
an	NN	O	O
explanation	NN	O	O
for	NN	O	O
many	NN	O	O
of	NN	O	O
the	NN	O	O
XP	NN	O	O
and	NN	O	O
CS	NN	O	O
symptoms	NN	O	O
that	NN	O	O
are	NN	O	O
difficult	NN	O	O
to	NN	O	O
rationalize	NN	O	O
based	NN	O	O
solely	NN	O	O
on	NN	O	O
an	NN	O	O
NER	NN	O	O
defect	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
this	NN	O	O
work	NN	O	O
presents	NN	O	O
the	NN	O	O
first	NN	O	O
detailed	NN	O	O
analysis	NN	O	O
of	NN	O	O
a	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
mutation	NN	O	O
in	NN	O	O
a	NN	O	O
basal	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
supports	NN	O	O
the	NN	O	O
concept	NN	O	O
that	NN	O	O
the	NN	O	O
combined	NN	O	O
XP/CS	NN	O	O
clinical	NN	O	O
entity	NN	O	O
is	NN	O	O
actually	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
combined	NN	O	O
transcription/repair	NN	O	O
deficiency	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Ets	NN	O	O
protein	NN	O	O
Spi-B	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
exclusively	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
development	NN	O	O
.	NN	O	O

Spi-B	NN	O	B-protein
and	NN	O	O
PU.1	NN	O	B-protein
are	NN	O	O
hematopoietic-specific	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
that	NN	O	O
constitute	NN	O	O
a	NN	O	O
subfamily	NN	O	O
of	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
contrary	NN	O	O
to	NN	O	O
previous	NN	O	O
reports	NN	O	O
,	NN	O	O
PU.1	NN	O	B-protein
and	NN	O	O
Spi-B	NN	O	B-protein
have	NN	O	O
very	NN	O	O
different	NN	O	O
expression	NN	O	O
patterns	NN	O	O
.	NN	O	O

PU.1	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
megakaryocytes	NN	O	B-cell_type
,	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
immature	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
at	NN	O	O
lower	NN	O	O
levels	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
.	NN	O	O

PU.1	NN	O	B-protein
is	NN	O	O
completely	NN	O	O
absent	NN	O	O
from	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
most	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
based	NN	O	O
on	NN	O	O
sensitive	NN	O	O
RT-PCR	NN	O	O
assays	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
Spi-B	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
exclusively	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
can	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
early	NN	O	O
fetal	NN	O	O
thymus	NN	O	O
and	NN	O	O
spleen	NN	O	O
.	NN	O	O

In	NN	O	O
situ	NN	O	O
hybridizations	NN	O	O
of	NN	O	O
adult	NN	O	O
murine	NN	O	O
tissues	NN	O	O
demonstrate	NN	O	O
Spi-B	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
medulla	NN	O	O
of	NN	O	O
the	NN	O	O
thymus	NN	O	O
,	NN	O	O
the	NN	O	O
white	NN	O	O
pulp	NN	O	O
of	NN	O	O
the	NN	O	O
spleen	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
germinal	NN	O	O
centers	NN	O	O
of	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
.	NN	O	O

Spi-B	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
very	NN	O	O
abundant	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
both	NN	O	O
Spi-B	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
are	NN	O	O
detected	NN	O	O
in	NN	O	O
some	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
and	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
suggest	NN	O	O
that	NN	O	O
Spi-B	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
increases	NN	O	O
during	NN	O	O
B	NN	O	O
cell	NN	O	O
maturation	NN	O	O
and	NN	O	O
decreases	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
maturation	NN	O	O
.	NN	O	O

Gel-retardation	NN	O	O
experiments	NN	O	O
show	NN	O	O
that	NN	O	O
Spi-B	NN	O	B-protein
can	NN	O	O
bind	NN	O	O
to	NN	O	O
all	NN	O	O
putative	NN	O	B-DNA
PU.1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
do	NN	O	O
not	NN	O	O
reveal	NN	O	O
any	NN	O	O
preferred	NN	O	O
Spi-B	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
both	NN	O	O
PU.1	NN	O	B-protein
and	NN	O	O
Spi-B	NN	O	B-protein
function	NN	O	O
as	NN	O	O
transcriptional	NN	O	O
activators	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
light-chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
E	NN	O	I-DNA
lambda	NN	O	I-DNA
2.4	NN	O	I-DNA
when	NN	O	O
coexpressed	NN	O	O
with	NN	O	O
Pip	NN	O	B-protein
(	NN	O	O
PU.1-interaction	NN	O	B-protein
partner	NN	O	I-protein
)	NN	O	O
in	NN	O	O
NIH-3T3	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
differences	NN	O	O
in	NN	O	O
patterns	NN	O	O
of	NN	O	O
expression	NN	O	O
between	NN	O	O
Spi-B	NN	O	B-protein
and	NN	O	O
PU.1	NN	O	B-protein
distinguish	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
each	NN	O	O
protein	NN	O	O
during	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
critical	NN	O	O
role	NN	O	O
of	NN	O	O
Sp1-	NN	O	B-protein
and	NN	O	I-protein
Ets-related	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
maintaining	NN	O	O
CTL-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
perforin	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
study	NN	O	O
was	NN	O	O
designed	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
potential	NN	O	O
cis-elements	NN	O	B-DNA
involved	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
perforin	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

DNase	NN	O	B-DNA
I	NN	O	I-DNA
hypersensitive	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
DHS	NN	O	B-DNA
)	NN	O	O
mapping	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
perforin	NN	O	B-DNA
locus	NN	O	I-DNA
contained	NN	O	O
six	NN	O	O
DHS	NN	O	B-DNA
within	NN	O	O
7.0	NN	O	O
kb	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
upstream	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
-7.0	NN	O	O
kb	NN	O	O
)	NN	O	O
and	NN	O	O
two	NN	O	O
DHS	NN	O	B-DNA
in	NN	O	O
intron	NN	O	B-DNA
2	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
six	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
upstream	NN	O	I-DNA
and	NN	O	O
one	NN	O	O
intronic	NN	O	B-DNA
DHS	NN	O	I-DNA
were	NN	O	O
detected	NN	O	O
in	NN	O	O
only	NN	O	O
perforin-expressing	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
(	NN	O	O
CAT	NN	O	B-protein
)	NN	O	O
activities	NN	O	O
directed	NN	O	O
by	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
upstream	NN	O	I-DNA
promoter	NN	O	I-DNA
were	NN	O	O
detected	NN	O	O
preferentially	NN	O	O
in	NN	O	O
perforin-expressing	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
construct	NN	O	O
termed	NN	O	O
PFP5a	NN	O	B-DNA
containing	NN	O	O
-795	NN	O	B-DNA
bp	NN	O	I-DNA
exhibited	NN	O	O
the	NN	O	O
highest	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
and	NN	O	O
PFP9a20	NN	O	B-DNA
containing	NN	O	O
only	NN	O	O
-73	NN	O	B-DNA
bp	NN	O	I-DNA
also	NN	O	O
produced	NN	O	O
significantly	NN	O	O
high	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
CTLL-R8	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
proximal	NN	O	B-DNA
region	NN	O	I-DNA
in	NN	O	O
PFP9a20	NN	O	B-DNA
contained	NN	O	O
two	NN	O	O
potential	NN	O	O
Sp1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
(	NN	O	O
GC	NN	O	B-DNA
box	NN	O	I-DNA
and	NN	O	O
GT	NN	O	B-DNA
box	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
one	NN	O	O
Ets	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
EBS	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
showed	NN	O	O
that	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
cis-elements	NN	O	B-DNA
bound	NN	O	O
specific	NN	O	O
protein	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

When	NN	O	O
single-point	NN	O	O
mutation	NN	O	O
was	NN	O	O
introduced	NN	O	O
to	NN	O	O
each	NN	O	O
GC	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
EBS	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
GT	NN	O	B-DNA
box	NN	O	I-DNA
in	NN	O	O
PFP9a20	NN	O	B-DNA
,	NN	O	O
at	NN	O	O
least	NN	O	O
3-fold	NN	O	O
less	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
CTLL-R8	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
confirm	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
perforin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
point	NN	O	O
mutation	NN	O	O
was	NN	O	O
introduced	NN	O	O
again	NN	O	O
to	NN	O	O
each	NN	O	O
proximal	NN	O	B-DNA
GC	NN	O	I-DNA
box	NN	O	I-DNA
,	NN	O	O
EBS	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
GT	NN	O	B-DNA
box	NN	O	I-DNA
of	NN	O	O
PFP5a	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
point	NN	O	O
mutations	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
2.5-	NN	O	O
to	NN	O	O
3-fold	NN	O	O
reduction	NN	O	O
of	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
proximal	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
may	NN	O	O
constitute	NN	O	O
a	NN	O	O
minimal	NN	O	O
region	NN	O	O
responsible	NN	O	O
for	NN	O	O
CTL-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
perforin	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Oct-2	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Oct-2	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
POU	NN	O	B-protein
(	NN	O	I-protein
Pit-Oct-Unc	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
only	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
neuronal	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

The	NN	O	O
primary	NN	O	B-RNA
RNA	NN	O	I-RNA
transcript	NN	O	I-RNA
of	NN	O	O
the	NN	O	O
gene	NN	O	O
is	NN	O	O
subject	NN	O	O
to	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
to	NN	O	O
yield	NN	O	O
different	NN	O	O
variants	NN	O	O
which	NN	O	O
can	NN	O	O
either	NN	O	O
activate	NN	O	O
or	NN	O	O
repress	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
forms	NN	O	O
produced	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
have	NN	O	O
a	NN	O	O
predominantly	NN	O	O
activating	NN	O	O
effect	NN	O	O
on	NN	O	O
gene	NN	O	O
expression	NN	O	O
whereas	NN	O	O
those	NN	O	O
produced	NN	O	O
in	NN	O	O
neuronal	NN	O	B-cell_type
cells	NN	O	I-cell_type
have	NN	O	O
a	NN	O	O
predominantly	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
and	NN	O	O
can	NN	O	O
repress	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
herpes	NN	O	B-DNA
simplex	NN	O	I-DNA
virus	NN	O	I-DNA
immediate-early	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
cellular	NN	O	B-DNA
tyrosine	NN	O	I-DNA
hydroxylase	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
Oct-2	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cellular	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
both	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
neuronal	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
viral	NN	O	O
latency	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cell-type-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
tumor	NN	O	I-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	O
ATF-2/JUN	NN	O	B-protein
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
tumor	NN	O	I-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
TNF-alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
earliest	NN	O	O
genes	NN	O	O
transcribed	NN	O	O
after	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
a	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
through	NN	O	O
its	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
or	NN	O	O
via	NN	O	O
the	NN	O	O
CD-40	NN	O	O
pathway	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
cases	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
can	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
immunosuppressants	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
,	NN	O	O
which	NN	O	O
suggested	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
proteins	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
two	NN	O	O
molecules	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
bind	NN	O	O
to	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
kappa	NN	O	I-DNA
3	NN	O	I-DNA
in	NN	O	O
association	NN	O	O
with	NN	O	O
ATF-2	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
proteins	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
an	NN	O	O
immediately	NN	O	O
adjacent	NN	O	O
cyclic	NN	O	B-DNA
AMP	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
CRE	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
,	NN	O	O
using	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
A20	NN	O	B-cell_line
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
a	NN	O	O
cell-type-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

In	NN	O	O
A20	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
regulated	NN	O	O
by	NN	O	O
NFATp	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
3	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Instead	NN	O	O
,	NN	O	O
ATF-2	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
proteins	NN	O	I-protein
bind	NN	O	O
to	NN	O	O
the	NN	O	O
composite	NN	O	B-DNA
kappa	NN	O	I-DNA
3/CRE	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
NFATp	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
a	NN	O	O
newly	NN	O	O
identified	NN	O	O
second	NN	O	O
NFAT	NN	O	B-DNA
site	NN	O	I-DNA
centered	NN	O	O
at	NN	O	O
-76	NN	O	B-DNA
nucleotides	NN	O	I-DNA
relative	NN	O	O
to	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
transcription	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
new	NN	O	O
site	NN	O	O
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
calcium-mediated	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A-sensitive	NN	O	O
induction	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
in	NN	O	O
both	NN	O	O
A20	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
Ar-5	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
quantitative	NN	O	O
DNase	NN	O	B-protein
footprinting	NN	O	O
of	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
using	NN	O	O
increasing	NN	O	O
amounts	NN	O	O
of	NN	O	O
recombinant	NN	O	O
NFATp	NN	O	B-protein
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
-76	NN	O	B-DNA
site	NN	O	I-DNA
binds	NN	O	O
to	NN	O	O
NFATp	NN	O	B-protein
with	NN	O	O
a	NN	O	O
higher	NN	O	O
affinity	NN	O	O
than	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
3	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Two	NN	O	O
other	NN	O	O
previously	NN	O	O
unrecognized	NN	O	O
NFATp-binding	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
proximal	NN	O	O
TNF-alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
were	NN	O	O
also	NN	O	O
identified	NN	O	O
by	NN	O	O
this	NN	O	O
analysis	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
through	NN	O	O
the	NN	O	O
differential	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
promoter	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
composite	NN	O	B-DNA
kappa	NN	O	I-DNA
3/CRE	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
a	NN	O	O
cell-type-specific	NN	O	O
manner	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
extracellular	NN	O	O
signal	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
Stat	NN	O	B-protein
5b	NN	O	I-protein
in	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
ErbB	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
sustained	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Self	NN	O	O
renewal	NN	O	O
of	NN	O	O
normal	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
is	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
c-ErbB	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
other	NN	O	O
receptors	NN	O	B-protein
(	NN	O	O
c-Kit/Epo-R	NN	O	B-protein
)	NN	O	O
regulate	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

To	NN	O	O
address	NN	O	O
possible	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
could	NN	O	O
explain	NN	O	O
this	NN	O	O
selective	NN	O	O
activity	NN	O	O
of	NN	O	O
c-ErbB	NN	O	B-protein
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
these	NN	O	O
receptors	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
different	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Stat	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Ligand	NN	O	O
activation	NN	O	O
of	NN	O	O
c-ErbB	NN	O	B-protein
induced	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
DNA-binding	NN	O	O
,	NN	O	O
and	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
of	NN	O	O
Stat	NN	O	B-protein
5b	NN	O	I-protein
in	NN	O	O
erythroblasts	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
ligand	NN	O	O
activation	NN	O	O
of	NN	O	O
c-Kit	NN	O	B-protein
was	NN	O	O
unable	NN	O	O
to	NN	O	O
induce	NN	O	O
any	NN	O	O
of	NN	O	O
these	NN	O	O
effects	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
cells	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
erythropoietin	NN	O	B-protein
receptor	NN	O	I-protein
caused	NN	O	O
specific	NN	O	O
DNA-binding	NN	O	O
of	NN	O	O
Stat	NN	O	B-protein
5b	NN	O	I-protein
,	NN	O	O
but	NN	O	O
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

These	NN	O	O
biochemical	NN	O	O
findings	NN	O	O
correlate	NN	O	O
perfectly	NN	O	O
with	NN	O	O
the	NN	O	O
selective	NN	O	O
ability	NN	O	O
of	NN	O	O
c-ErbB	NN	O	B-protein
to	NN	O	O
cause	NN	O	O
sustained	NN	O	O
self	NN	O	O
renewal	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Defective	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
associated	NN	O	O
with	NN	O	O
impaired	NN	O	O
expression	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
FosB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
JunB	NN	O	B-protein
in	NN	O	O
anergic	NN	O	B-cell_line
T	NN	O	I-cell_line
helper	NN	O	I-cell_line
1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Anergic	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
Th	NN	O	I-cell_line
cells	NN	O	I-cell_line
do	NN	O	O
not	NN	O	O
produce	NN	O	O
IL-2	NN	O	B-protein
when	NN	O	O
challenged	NN	O	O
with	NN	O	O
Ag-pulsed	NN	O	B-cell_line
accessory	NN	O	I-cell_line
cells	NN	O	I-cell_line
because	NN	O	O
of	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
defect	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
work	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
these	NN	O	O
anergic	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
defective	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
up-regulate	NN	O	O
protein	NN	O	O
binding	NN	O	O
and	NN	O	O
transactivation	NN	O	O
at	NN	O	O
two	NN	O	O
critical	NN	O	O
IL-2	NN	O	B-DNA
DNA	NN	O	I-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
:	NN	O	O
NF-AT	NN	O	B-protein
(	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
;	NN	O	O
a	NN	O	O
sequence	NN	O	O
that	NN	O	O
binds	NN	O	O
a	NN	O	O
heterotrimeric	NN	O	B-protein
NFATp	NN	O	I-protein
,	NN	O	I-protein
Fos	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
Jun	NN	O	I-protein
protein	NN	O	I-protein
complex	NN	O	I-protein
)	NN	O	O
and	NN	O	O
Activator	NN	O	B-protein
Protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
(	NN	O	O
that	NN	O	O
binds	NN	O	O
Fos	NN	O	B-protein
and	NN	O	I-protein
Jun	NN	O	I-protein
heterodimers	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
impaired	NN	O	O
DNA-protein	NN	O	O
interactions	NN	O	O
in	NN	O	O
anergic	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
associated	NN	O	O
with	NN	O	O
poor	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	O
AP-1	NN	O	B-protein
family	NN	O	O
members	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
FosB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
JunB	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
reduced	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
was	NN	O	O
not	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
global	NN	O	O
TCR/CD3	NN	O	B-protein
-signaling	NN	O	O
defect	NN	O	O
because	NN	O	O
CD3	NN	O	B-protein
cross-linking	NN	O	O
induced	NN	O	O
an	NN	O	O
equivalent	NN	O	O
increase	NN	O	O
in	NN	O	O
intracellular-free	NN	O	O
calcium	NN	O	O
ions	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
NFATp	NN	O	B-protein
dephosphorylation	NN	O	O
,	NN	O	O
translocation	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
in	NN	O	O
both	NN	O	O
normal	NN	O	B-cell_type
and	NN	O	O
anergic	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
defective	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
due	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
selective	NN	O	O
block	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
Fos	NN	O	I-protein
and	NN	O	I-protein
Jun	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
in	NN	O	O
anergic	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Sequence-specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	B-protein
coactivator	NN	O	I-protein
OCA-B	NN	O	B-protein
.	NN	O	O

B-cell-specific	NN	O	O
transcription	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
a	NN	O	O
POU	NN	O	B-DNA
domain	NN	O	I-DNA
containing	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Oct-1	NN	O	B-protein
or	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
with	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	B-protein
coactivator	NN	O	I-protein
OCA-B	NN	O	B-protein
(	NN	O	O
Bob-1	NN	O	B-protein
,	NN	O	O
OBF-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
a	NN	O	O
prototype	NN	O	B-DNA
octamer	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
OCA-B	NN	O	B-protein
binds	NN	O	O
DNA	NN	O	O
directly	NN	O	O
in	NN	O	O
the	NN	O	O
major	NN	O	O
groove	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
subdomains	NN	O	O
of	NN	O	O
the	NN	O	O
POU	NN	O	B-DNA
domain	NN	O	I-DNA
,	NN	O	O
requiring	NN	O	O
both	NN	O	O
an	NN	O	O
A	NN	O	O
at	NN	O	O
the	NN	O	O
fifth	NN	O	B-DNA
position	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
element	NN	O	I-DNA
and	NN	O	O
contact	NN	O	O
with	NN	O	O
the	NN	O	O
POU	NN	O	B-DNA
domain	NN	O	I-DNA
.	NN	O	O

An	NN	O	O
amino-terminal	NN	O	B-protein
fragment	NN	O	I-protein
of	NN	O	O
OCA-B	NN	O	B-protein
binds	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
POU	NN	O	B-DNA
domain	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
same	NN	O	O
sequence	NN	O	O
specificity	NN	O	O
.	NN	O	O

Coactivator	NN	O	O
OCA-B	NN	O	B-protein
may	NN	O	O
undergo	NN	O	O
a	NN	O	O
POU-dependent	NN	O	O
conformational	NN	O	O
change	NN	O	O
that	NN	O	O
exposes	NN	O	O
its	NN	O	O
amino	NN	O	O
terminus	NN	O	O
,	NN	O	O
allowing	NN	O	O
it	NN	O	O
to	NN	O	O
recognize	NN	O	O
specific	NN	O	O
DNA	NN	O	B-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
major	NN	O	O
groove	NN	O	O
within	NN	O	O
the	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
Oct-1	NN	O	B-protein
or	NN	O	O
Oct-2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
recognition	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
POU	NN	O	B-DNA
domain	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
sequence	NN	O	I-DNA
by	NN	O	O
OCA-B	NN	O	B-protein
provides	NN	O	O
a	NN	O	O
mechanism	NN	O	O
for	NN	O	O
differential	NN	O	O
regulation	NN	O	O
of	NN	O	O
octamer	NN	O	B-DNA
sites	NN	O	I-DNA
containing	NN	O	O
genes	NN	O	O
by	NN	O	O
the	NN	O	O
ubiquitous	NN	O	B-protein
factor	NN	O	I-protein
Oct-1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Uneven	NN	O	O
X	NN	O	B-DNA
inactivation	NN	O	O
in	NN	O	O
a	NN	O	O
female	NN	O	O
monozygotic	NN	O	O
twin	NN	O	O
pair	NN	O	O
with	NN	O	O
Fabry	NN	O	O
disease	NN	O	O
and	NN	O	O
discordant	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
alpha-galactosidase	NN	O	B-DNA
A	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
describe	NN	O	O
two	NN	O	O
female	NN	O	O
monozygotic	NN	O	O
(	NN	O	O
MZ	NN	O	O
)	NN	O	O
twins	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
Fabry	NN	O	O
disease	NN	O	O
,	NN	O	O
an	NN	O	O
X	NN	O	O
linked	NN	O	O
disorder	NN	O	O
resulting	NN	O	O
from	NN	O	O
the	NN	O	O
deficient	NN	O	O
activity	NN	O	O
of	NN	O	O
alpha-galactosidase	NN	O	B-protein
A	NN	O	I-protein
.	NN	O	O

While	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
twins	NN	O	O
was	NN	O	O
clinically	NN	O	O
affected	NN	O	O
,	NN	O	O
the	NN	O	O
other	NN	O	O
was	NN	O	O
asymptomatic	NN	O	O
.	NN	O	O

Enzymatic	NN	O	O
assay	NN	O	O
of	NN	O	O
alpha-galactosidase	NN	O	B-protein
in	NN	O	O
blood	NN	O	B-cell_type
leucocytes	NN	O	I-cell_type
,	NN	O	O
skin	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
,	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus	NN	O	I-cell_line
transformed	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
hair	NN	O	O
follicles	NN	O	O
of	NN	O	O
the	NN	O	O
twins	NN	O	O
and	NN	O	O
their	NN	O	O
parents	NN	O	O
confirmed	NN	O	O
the	NN	O	O
heterozygous	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
twins	NN	O	O
and	NN	O	O
indicated	NN	O	O
that	NN	O	O
Fabry	NN	O	O
disease	NN	O	O
had	NN	O	O
occurred	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
de	NN	O	O
novo	NN	O	O
mutation	NN	O	O
.	NN	O	O

The	NN	O	O
son	NN	O	O
of	NN	O	O
the	NN	O	O
unaffected	NN	O	O
twin	NN	O	O
sister	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
hemizygous	NN	O	O
.	NN	O	O

Molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
alpha-galactosidase	NN	O	B-DNA
A	NN	O	I-DNA
gene	NN	O	I-DNA
permitted	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
an	NN	O	O
as	NN	O	O
yet	NN	O	O
undescribed	NN	O	O
point	NN	O	O
mutation	NN	O	O
at	NN	O	O
position	NN	O	B-DNA
10182	NN	O	I-DNA
of	NN	O	I-DNA
exon	NN	O	I-DNA
5	NN	O	I-DNA
which	NN	O	O
causes	NN	O	O
an	NN	O	O
Asp	NN	O	O
to	NN	O	O
Asn	NN	O	O
substitution	NN	O	O
at	NN	O	O
codon	NN	O	B-DNA
231	NN	O	I-DNA
.	NN	O	O

Single	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
analysis	NN	O	O
again	NN	O	O
showed	NN	O	O
the	NN	O	O
heterozygous	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
twins	NN	O	O
and	NN	O	O
a	NN	O	O
normal	NN	O	O
pattern	NN	O	O
in	NN	O	O
their	NN	O	O
parents	NN	O	O
.	NN	O	O

The	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
discordant	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
d	NN	O	O
novo	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
twins	NN	O	O
was	NN	O	O
investigated	NN	O	O
by	NN	O	O
studying	NN	O	O
their	NN	O	O
X	NN	O	O
inactivation	NN	O	O
status	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
inactive	NN	O	B-DNA
X	NN	O	I-DNA
specific	NN	O	O
methylation	NN	O	O
at	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
showed	NN	O	O
unbalanced	NN	O	O
inactivation	NN	O	O
in	NN	O	O
the	NN	O	O
twins	NN	O	O
'	NN	O	O
fibroblasts	NN	O	O
and	NN	O	O
in	NN	O	O
opposite	NN	O	O
directions	NN	O	O
.	NN	O	O

While	NN	O	O
the	NN	O	O
maternally	NN	O	B-DNA
derived	NN	O	I-DNA
X	NN	O	I-DNA
chromosome	NN	O	I-DNA
was	NN	O	O
preferentially	NN	O	O
active	NN	O	O
in	NN	O	O
the	NN	O	O
asymptomatic	NN	O	O
twin	NN	O	O
,	NN	O	O
the	NN	O	O
paternal	NN	O	B-DNA
X	NN	O	I-DNA
chromosome	NN	O	I-DNA
was	NN	O	O
active	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
,	NN	O	O
affected	NN	O	O
twin	NN	O	O
and	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
her	NN	O	O
hemizygotic	NN	O	O
nephew	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
paternal	NN	O	B-DNA
X	NN	O	I-DNA
chromosome	NN	O	I-DNA
carries	NN	O	O
the	NN	O	O
de	NN	O	O
novo	NN	O	O
alpha-galactosidase	NN	O	B-protein
A	NN	O	I-protein
mutation	NN	O	O
and	NN	O	O
that	NN	O	O
uneven	NN	O	O
X	NN	O	O
inactivation	NN	O	O
is	NN	O	O
the	NN	O	O
underlying	NN	O	O
mechanism	NN	O	O
for	NN	O	O
disease	NN	O	O
expression	NN	O	O
in	NN	O	O
this	NN	O	O
novel	NN	O	O
female	NN	O	O
MZ	NN	O	O
twin	NN	O	O
pair	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
documented	NN	O	O
case	NN	O	O
of	NN	O	O
female	NN	O	O
twins	NN	O	O
discordant	NN	O	O
for	NN	O	O
Fabry	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Precise	NN	O	O
alignment	NN	O	B-DNA
of	NN	O	I-DNA
sites	NN	O	I-DNA
required	NN	O	O
for	NN	O	O
mu	NN	O	B-DNA
enhancer	NN	O	I-DNA
activation	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
lymphocyte-specific	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
mu	NN	O	I-DNA
heavy-chain	NN	O	I-DNA
gene	NN	O	I-DNA
intronic	NN	O	I-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
multiple	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
previously	NN	O	O
defined	NN	O	O
minimal	NN	O	B-DNA
enhancer	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
muA	NN	O	B-DNA
,	NN	O	I-DNA
muE3	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
muB	NN	O	I-DNA
sites	NN	O	I-DNA
is	NN	O	O
transactivated	NN	O	O
by	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
the	NN	O	O
ETS-domain	NN	O	B-protein
proteins	NN	O	I-protein
PU.1	NN	O	B-protein
and	NN	O	O
Ets-1	NN	O	B-protein
in	NN	O	O
nonlymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
core	NN	O	O
GGAAs	NN	O	O
of	NN	O	O
the	NN	O	O
muA	NN	O	B-DNA
and	NN	O	I-DNA
muB	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
separated	NN	O	O
by	NN	O	O
30	NN	O	O
nucleotides	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
ETS	NN	O	B-protein
proteins	NN	O	I-protein
bind	NN	O	O
to	NN	O	O
these	NN	O	O
sites	NN	O	O
from	NN	O	O
these	NN	O	O
same	NN	O	O
side	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
helix	NN	O	O
.	NN	O	O

We	NN	O	O
tested	NN	O	O
the	NN	O	O
necessity	NN	O	O
for	NN	O	O
appropriate	NN	O	O
spatial	NN	O	O
alignment	NN	O	O
of	NN	O	O
these	NN	O	O
elements	NN	O	B-DNA
by	NN	O	O
using	NN	O	O
mutated	NN	O	B-DNA
enhancers	NN	O	I-DNA
with	NN	O	O
altered	NN	O	O
spacings	NN	O	O
.	NN	O	O

A	NN	O	O
4-	NN	O	O
or	NN	O	O
10-bp	NN	O	O
insertion	NN	O	O
between	NN	O	O
muE3	NN	O	B-DNA
and	NN	O	O
muB	NN	O	B-DNA
inactivated	NN	O	O
the	NN	O	O
mu	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
S194	NN	O	B-cell_line
plasma	NN	O	I-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
in	NN	O	O
vitro	NN	O	O
binding	NN	O	O
of	NN	O	O
Ets-1	NN	O	O
,	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
or	NN	O	O
the	NN	O	O
muE3-binding	NN	O	B-protein
protein	NN	O	I-protein
TFE3	NN	O	B-protein
,	NN	O	O
alone	NN	O	O
or	NN	O	O
in	NN	O	O
pairwise	NN	O	O
combinations	NN	O	O
.	NN	O	O

Circular	NN	O	O
permutation	NN	O	O
and	NN	O	O
phasing	NN	O	O
analyses	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
PU.1	NN	O	B-protein
binding	NN	O	O
but	NN	O	O
not	NN	O	O
TFE3	NN	O	B-protein
or	NN	O	O
Ets-1	NN	O	B-protein
bends	NN	O	O
mu	NN	O	B-DNA
enhancer	NN	O	I-DNA
DNA	NN	O	I-DNA
toward	NN	O	O
the	NN	O	O
major	NN	O	O
groove	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
requirement	NN	O	O
for	NN	O	O
precise	NN	O	O
spacing	NN	O	O
of	NN	O	O
the	NN	O	O
muA	NN	O	B-DNA
and	NN	O	I-DNA
muB	NN	O	I-DNA
elements	NN	O	I-DNA
is	NN	O	O
due	NN	O	O
in	NN	O	O
part	NN	O	O
to	NN	O	O
a	NN	O	O
directed	NN	O	O
DNA	NN	O	O
bend	NN	O	O
induced	NN	O	O
by	NN	O	O
PU.1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Cortisone-resistant	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
]	NN	O	O

There	NN	O	O
is	NN	O	O
general	NN	O	O
agreement	NN	O	O
on	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
:	NN	O	O
an	NN	O	O
accumulation	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
,	NN	O	O
degranulated	NN	O	B-cell_type
mast	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
very	NN	O	O
severe	NN	O	O
forms	NN	O	O
,	NN	O	O
granulocytes	NN	O	B-cell_type
has	NN	O	O
constantly	NN	O	O
been	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
bronchial	NN	O	O
mucosa	NN	O	O
.	NN	O	O

In	NN	O	O
allergic	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
,	NN	O	O
inflammation	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
orchestrated	NN	O	O
predominantly	NN	O	O
by	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
with	NN	O	O
a	NN	O	O
phenotype	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
Th2	NN	O	B-cell_type
subset	NN	O	I-cell_type
able	NN	O	O
to	NN	O	O
produce	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-5	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
corticosteroids	NN	O	O
are	NN	O	O
the	NN	O	O
most	NN	O	O
potent	NN	O	O
therapeutic	NN	O	O
agents	NN	O	O
used	NN	O	O
for	NN	O	O
this	NN	O	O
disease	NN	O	O
,	NN	O	O
their	NN	O	O
anti-inflammatory	NN	O	O
effect	NN	O	O
differs	NN	O	O
from	NN	O	O
patient	NN	O	O
to	NN	O	O
patient	NN	O	O
.	NN	O	O

Some	NN	O	O
criteria	NN	O	O
which	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
define	NN	O	O
steroid-resistant	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
are	NN	O	O
listed	NN	O	O
here	NN	O	O
.	NN	O	O

This	NN	O	O
review	NN	O	O
analyzes	NN	O	O
various	NN	O	O
molecular	NN	O	O
alterations	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
deficient	NN	O	O
response	NN	O	O
to	NN	O	O
corticosteroid	NN	O	O
treatment	NN	O	O
observed	NN	O	O
in	NN	O	O
steroid-resistant	NN	O	O
bronchial	NN	O	O
asthmatic	NN	O	O
subjects	NN	O	O
.	NN	O	O

New	NN	O	O
knowledge	NN	O	O
on	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
steroid	NN	O	O
resistance	NN	O	O
may	NN	O	O
have	NN	O	O
important	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
chronic	NN	O	O
asthma	NN	O	O
and	NN	O	O
other	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mechanisms	NN	O	O
of	NN	O	O
transactivation	NN	O	O
by	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells-1	NN	O	I-protein
.	NN	O	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells-family	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
NFAT1/NFATp	NN	O	B-protein
,	NN	O	O
NFATc	NN	O	B-protein
,	NN	O	O
NFAT3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NFAT4/NFATx/NFATc3	NN	O	B-protein
)	NN	O	O
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
other	NN	O	O
genes	NN	O	O
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
defined	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
transactivation	NN	O	O
by	NN	O	O
NFAT1	NN	O	B-protein
.	NN	O	O

NFAT1	NN	O	B-protein
possesses	NN	O	O
two	NN	O	O
transactivation	NN	O	B-DNA
domains	NN	O	I-DNA
whose	NN	O	O
sequences	NN	O	O
are	NN	O	O
not	NN	O	O
conserved	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
NFAT-family	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
conserved	NN	O	O
DNA-binding	NN	O	B-DNA
domain	NN	O	I-DNA
that	NN	O	O
mediates	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
cooperating	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
even	NN	O	O
when	NN	O	O
it	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
other	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
NH2-terminal	NN	O	B-protein
transactivation	NN	O	I-protein
domain	NN	O	I-protein
is	NN	O	O
modulated	NN	O	O
by	NN	O	O
an	NN	O	O
adjacent	NN	O	O
regulatory	NN	O	B-protein
region	NN	O	I-protein
that	NN	O	O
contains	NN	O	O
several	NN	O	O
conserved	NN	O	B-DNA
sequence	NN	O	I-DNA
motifs	NN	O	I-DNA
represented	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
emphasize	NN	O	O
the	NN	O	O
multiple	NN	O	O
levels	NN	O	O
at	NN	O	O
which	NN	O	O
NFAT-dependent	NN	O	O
transactivation	NN	O	O
is	NN	O	O
regulated	NN	O	O
,	NN	O	O
and	NN	O	O
predict	NN	O	O
significant	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
architecture	NN	O	O
of	NN	O	O
cooperative	NN	O	O
transcription	NN	O	B-protein
complexes	NN	O	I-protein
containing	NN	O	O
different	NN	O	O
NFAT-family	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
activator	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
AP	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
by	NN	O	O
CD28	NN	O	B-protein
stimulation	NN	O	O
involves	NN	O	O
both	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
and	NN	O	O
acidic	NN	O	B-protein
sphingomyelinase	NN	O	I-protein
signals	NN	O	O
.	NN	O	O

A	NN	O	O
major	NN	O	O
obstacle	NN	O	O
in	NN	O	O
understanding	NN	O	O
the	NN	O	O
signaling	NN	O	O
events	NN	O	O
that	NN	O	O
follow	NN	O	O
CD28	NN	O	B-protein
receptor	NN	O	O
ligation	NN	O	O
arises	NN	O	O
from	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
CD28	NN	O	B-protein
acts	NN	O	O
as	NN	O	O
a	NN	O	O
costimulus	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
engagement	NN	O	O
,	NN	O	O
making	NN	O	O
it	NN	O	O
difficult	NN	O	O
to	NN	O	O
assess	NN	O	O
the	NN	O	O
relative	NN	O	O
contribution	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
signals	NN	O	O
as	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
.	NN	O	O

To	NN	O	O
overcome	NN	O	O
this	NN	O	O
problem	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
exploited	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
activated	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
blasts	NN	O	I-cell_type
can	NN	O	O
be	NN	O	O
stimulated	NN	O	O
via	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
surface	NN	O	I-protein
molecule	NN	O	I-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
antigenic	NN	O	O
challenge	NN	O	O
;	NN	O	O
thus	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
been	NN	O	O
able	NN	O	O
to	NN	O	O
observe	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
CD28	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
isolation	NN	O	O
.	NN	O	O

Using	NN	O	O
this	NN	O	O
system	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
CD28	NN	O	B-protein
stimulation	NN	O	O
by	NN	O	O
B7-transfected	NN	O	B-cell_line
CHO	NN	O	I-cell_line
cells	NN	O	I-cell_line
induced	NN	O	O
a	NN	O	O
proliferative	NN	O	O
response	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
was	NN	O	O
not	NN	O	O
accompanied	NN	O	O
by	NN	O	O
measurable	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
subsequent	NN	O	O
analysis	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
generation	NN	O	O
revealed	NN	O	O
that	NN	O	O
B7	NN	O	O
stimulation	NN	O	O
induced	NN	O	O
both	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	I-protein
NF-kappaB	NN	O	I-protein
)	NN	O	I-protein
complexes	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
engagement	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
by	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
MHC/superantigen	NN	O	I-protein
,	NN	O	O
either	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
CD28	NN	O	B-protein
ligation	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Using	NN	O	O
selective	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

This	NN	O	O
revealed	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
generation	NN	O	O
was	NN	O	O
sensitive	NN	O	O
to	NN	O	O
chloroquine	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
acidic	NN	O	B-protein
sphingomyelinase	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
to	NN	O	O
the	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
inhibitor	NN	O	O
,	NN	O	O
wortmannin	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
AP-1	NN	O	B-protein
generation	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
wortmannin	NN	O	O
and	NN	O	O
was	NN	O	O
also	NN	O	O
variably	NN	O	O
sensitive	NN	O	O
to	NN	O	O
chloroquine	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
in	NN	O	O
activated	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
CD28	NN	O	B-protein
-derived	NN	O	O
signals	NN	O	O
can	NN	O	O
stimulate	NN	O	O
proliferation	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
via	NN	O	O
NF-kappaB	NN	O	O
and	NN	O	O
AP-1	NN	O	O
generation	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
response	NN	O	O
uses	NN	O	O
both	NN	O	O
acidic	NN	O	B-protein
sphingomyelinase	NN	O	I-protein
and	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
-linked	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

Presence	NN	O	O
of	NN	O	O
a	NN	O	O
variant	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
normal	NN	O	O
individuals	NN	O	O
and	NN	O	O
lupus	NN	O	O
patients	NN	O	O
.	NN	O	O

Estrogen	NN	O	O
may	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
(	NN	O	O
SLE	NN	O	O
)	NN	O	O
via	NN	O	O
its	NN	O	O
intracellular	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
various	NN	O	O
isoforms	NN	O	O
of	NN	O	O
the	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
in	NN	O	O
SLE	NN	O	O
we	NN	O	O
isolated	NN	O	O
RNA	NN	O	O
from	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
lupus	NN	O	O
patients	NN	O	O
and	NN	O	O
normal	NN	O	O
controls	NN	O	O
.	NN	O	O

Using	NN	O	O
RT-PCR	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
identify	NN	O	O
both	NN	O	O
the	NN	O	O
full	NN	O	B-protein
length	NN	O	I-protein
wild-type	NN	O	I-protein
form	NN	O	I-protein
and	NN	O	O
an	NN	O	O
isoform	NN	O	O
of	NN	O	O
the	NN	O	O
ER	NN	O	B-protein
which	NN	O	O
precisely	NN	O	O
lacks	NN	O	O
exon	NN	O	B-DNA
V	NN	O	I-DNA
in	NN	O	O
both	NN	O	O
patient	NN	O	O
and	NN	O	O
normal	NN	O	O
individuals	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
,	NN	O	O
although	NN	O	O
limited	NN	O	O
,	NN	O	O
suggest	NN	O	O
that	NN	O	O
normal	NN	O	O
individuals	NN	O	O
can	NN	O	O
express	NN	O	O
both	NN	O	O
the	NN	O	O
wild-type	NN	O	O
and	NN	O	O
truncated	NN	O	O
version	NN	O	O
at	NN	O	O
the	NN	O	O
same	NN	O	O
time	NN	O	O
,	NN	O	O
whereas	NN	O	O
lupus	NN	O	O
patients	NN	O	O
only	NN	O	O
express	NN	O	O
either	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
or	NN	O	I-protein
the	NN	O	I-protein
truncated	NN	O	I-protein
ER	NN	O	I-protein
.	NN	O	O

This	NN	O	O
finding	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
reasons	NN	O	O
for	NN	O	O
the	NN	O	O
prevalence	NN	O	O
of	NN	O	O
lupus	NN	O	O
in	NN	O	O
females	NN	O	O
and	NN	O	O
of	NN	O	O
the	NN	O	O
estrogenic	NN	O	O
effects	NN	O	O
on	NN	O	O
SLE	NN	O	O
disease	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
sialoadhesin	NN	O	B-protein
expression	NN	O	O
on	NN	O	O
rat	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Induction	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
enhancement	NN	O	O
by	NN	O	O
IFN-beta	NN	O	B-protein
,	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
lipopolysaccharide	NN	O	O
.	NN	O	O

Sialoadhesin	NN	O	B-protein
is	NN	O	O
a	NN	O	O
macrophage-restricted	NN	O	B-protein
member	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Ig	NN	O	B-protein
superfamily	NN	O	I-protein
that	NN	O	O
mediates	NN	O	O
adhesion	NN	O	O
with	NN	O	O
lymphoid	NN	O	B-cell_type
and	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
is	NN	O	O
expressed	NN	O	O
on	NN	O	O
a	NN	O	O
subpopulation	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
and	NN	O	O
in	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
(	NN	O	O
e.g.	NN	O	O
,	NN	O	O
during	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
sialoadhesin	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
show	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
induce	NN	O	O
sialoadhesin	NN	O	B-protein
expression	NN	O	O
on	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
rat	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
rat	NN	O	B-cell_line
macrophage	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
R2	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
cytokines	NN	O	B-protein
IFN-beta	NN	O	B-protein
,	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
LPS	NN	O	O
,	NN	O	O
although	NN	O	O
unable	NN	O	O
to	NN	O	O
induce	NN	O	O
sialoadhesin	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
themselves	NN	O	O
,	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
enhance	NN	O	O
GC-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
sialoadhesin	NN	O	B-protein
.	NN	O	O

Sialoadhesin	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
functional	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
assays	NN	O	O
with	NN	O	O
human	NN	O	B-cell_type
RBCs	NN	O	I-cell_type
.	NN	O	O

Northern	NN	O	O
blotting	NN	O	O
experiments	NN	O	O
indicated	NN	O	O
that	NN	O	O
regulation	NN	O	O
predominantly	NN	O	O
occurred	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
the	NN	O	O
different	NN	O	O
combinations	NN	O	O
of	NN	O	O
GC	NN	O	O
and	NN	O	O
cytokines	NN	O	B-protein
/LPS	NN	O	O
revealed	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
GC-dependent	NN	O	O
enhancement	NN	O	O
of	NN	O	O
sialoadhesin	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
with	NN	O	O
IFN-beta	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
being	NN	O	O
more	NN	O	O
potent	NN	O	O
than	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
LPS	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
LPS	NN	O	O
could	NN	O	O
be	NN	O	O
reproduced	NN	O	O
by	NN	O	O
priming	NN	O	O
,	NN	O	O
whereas	NN	O	O
IFN-beta	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
were	NN	O	O
required	NN	O	O
simultaneously	NN	O	O
with	NN	O	O
GC	NN	O	O
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
sialoadhesin	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
GC	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
not	NN	O	O
by	NN	O	O
mineralocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
inhibition	NN	O	O
experiments	NN	O	O
with	NN	O	O
specific	NN	O	O
antagonists	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
the	NN	O	O
adrenal	NN	O	O
gland	NN	O	O
,	NN	O	O
the	NN	O	O
major	NN	O	O
site	NN	O	O
of	NN	O	O
endogenous	NN	O	O
GC	NN	O	O
production	NN	O	O
,	NN	O	O
express	NN	O	O
sialoadhesin	NN	O	B-protein
.	NN	O	O

This	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
GC	NN	O	O
act	NN	O	O
as	NN	O	O
a	NN	O	O
primary	NN	O	O
inducer	NN	O	O
of	NN	O	O
sialoadhesin	NN	O	B-protein
expression	NN	O	O
on	NN	O	O
rat	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
response	NN	O	O
can	NN	O	O
be	NN	O	O
enhanced	NN	O	O
by	NN	O	O
IFN-beta	NN	O	B-protein
,	NN	O	O
T	NN	O	B-protein
cell-derived	NN	O	I-protein
cytokines	NN	O	I-protein
,	NN	O	O
or	NN	O	O
LPS	NN	O	O
.	NN	O	O

-DOCSTART-	O

Apoptosis	NN	O	O
mediated	NN	O	O
by	NN	O	O
HIV	NN	O	B-protein
protease	NN	O	I-protein
is	NN	O	O
preceded	NN	O	O
by	NN	O	O
cleavage	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
immunodeficiency	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
HIV	NN	O	I-protein
)	NN	O	I-protein
protease	NN	O	I-protein
in	NN	O	O
cultured	NN	O	B-cell_line
cells	NN	O	I-cell_line
leads	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
preceded	NN	O	O
by	NN	O	O
cleavage	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
,	NN	O	O
a	NN	O	O
key	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
protects	NN	O	O
cells	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
from	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
protease	NN	O	I-protein
and	NN	O	O
prevents	NN	O	O
cell	NN	O	O
death	NN	O	O
following	NN	O	O
HIV	NN	O	O
infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
reducing	NN	O	O
the	NN	O	O
yields	NN	O	O
of	NN	O	O
viral	NN	O	B-protein
structural	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
infectivity	NN	O	O
,	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

We	NN	O	O
present	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
protease	NN	O	I-protein
depletes	NN	O	O
the	NN	O	O
infected	NN	O	O
cells	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
,	NN	O	O
leading	NN	O	O
to	NN	O	O
oxidative	NN	O	O
stress-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
cellular	NN	O	B-protein
factor	NN	O	I-protein
required	NN	O	O
for	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
ultimately	NN	O	O
to	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Purified	NN	O	O
bcl-2	NN	O	B-protein
is	NN	O	O
cleaved	NN	O	O
by	NN	O	O
HIV	NN	O	B-protein
protease	NN	O	I-protein
between	NN	O	O
phenylalanine	NN	O	O
112	NN	O	O
and	NN	O	O
alanine	NN	O	O
113	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
new	NN	O	O
option	NN	O	O
for	NN	O	O
HIV	NN	O	O
gene	NN	O	O
therapy	NN	O	O
;	NN	O	O
bcl-2	NN	O	B-protein
muteins	NN	O	I-protein
that	NN	O	O
have	NN	O	O
noncleavable	NN	O	B-protein
alterations	NN	O	I-protein
surrounding	NN	O	O
the	NN	O	O
HIV	NN	O	B-protein
protease	NN	O	I-protein
cleavage	NN	O	I-protein
site	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Alpha	NN	O	B-protein
4	NN	O	I-protein
beta	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
CD49d/CD29	NN	O	I-protein
)	NN	O	I-protein
integrin	NN	O	I-protein
costimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
enhances	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
and	NN	O	O
cytokine	NN	O	O
induction	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
altered	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
anti-CD3	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
integrin	NN	O	B-protein
alpha	NN	O	I-protein
4	NN	O	I-protein
beta	NN	O	I-protein
1	NN	O	I-protein
can	NN	O	O
provide	NN	O	O
a	NN	O	O
costimulus	NN	O	O
to	NN	O	O
induce	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
and	NN	O	O
IL-2R	NN	O	B-protein
expression	NN	O	O
leading	NN	O	O
to	NN	O	O
enhanced	NN	O	O
proliferation	NN	O	O
of	NN	O	O
purified	NN	O	O
,	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
recombinant	NN	O	B-protein
vascular-cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
,	NN	O	O
when	NN	O	O
co-immobilized	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
,	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
NF-AT	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
as	NN	O	O
determined	NN	O	O
by	NN	O	O
electromobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

alpha	NN	O	B-protein
4	NN	O	I-protein
beta	NN	O	I-protein
1	NN	O	I-protein
ligation	NN	O	O
alone	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
.	NN	O	O

The	NN	O	O
requirements	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
reflected	NN	O	O
the	NN	O	O
requirements	NN	O	O
for	NN	O	O
the	NN	O	O
secretion	NN	O	O
of	NN	O	O
multiple	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
including	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
granulocyte	NN	O	B-protein
macrophage-CSF	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
in	NN	O	O
vitro-cultured	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
did	NN	O	O
not	NN	O	O
require	NN	O	O
costimulation	NN	O	O
for	NN	O	O
cytokine	NN	O	B-protein
secretion	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
anti-CD3	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
the	NN	O	O
dose	NN	O	O
response	NN	O	O
to	NN	O	O
anti-CD3	NN	O	B-protein
stimulation	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
half-maximal	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
was	NN	O	O
achieved	NN	O	O
using	NN	O	O
the	NN	O	O
same	NN	O	O
dose	NN	O	O
of	NN	O	O
anti-CD3	NN	O	B-protein
for	NN	O	O
both	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
and	NN	O	O
cultured	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
dose	NN	O	O
of	NN	O	O
OKT3	NN	O	B-protein
required	NN	O	O
to	NN	O	O
achieve	NN	O	O
half-maximal	NN	O	O
activation	NN	O	O
was	NN	O	O
the	NN	O	O
same	NN	O	O
using	NN	O	O
PMA	NN	O	O
or	NN	O	O
different	NN	O	O
concentrations	NN	O	O
of	NN	O	O
alpha	NN	O	B-protein
4	NN	O	I-protein
beta	NN	O	I-protein
1	NN	O	I-protein
ligands	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
costimulation	NN	O	O
by	NN	O	O
alpha	NN	O	B-protein
4	NN	O	I-protein
beta	NN	O	I-protein
1	NN	O	I-protein
ligands	NN	O	O
was	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
stabilization	NN	O	O
of	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
its	NN	O	O
substrate	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
,	NN	O	O
rather	NN	O	O
,	NN	O	O
that	NN	O	O
alpha	NN	O	B-protein
4	NN	O	I-protein
beta	NN	O	I-protein
1	NN	O	I-protein
in	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
delivers	NN	O	O
a	NN	O	O
distinct	NN	O	O
signal	NN	O	O
that	NN	O	O
synergizes	NN	O	O
early	NN	O	O
with	NN	O	O
signals	NN	O	O
initiated	NN	O	O
by	NN	O	O
TCR/CD3	NN	O	B-protein
ligation	NN	O	O
to	NN	O	O
induce	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
multiple	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
required	NN	O	O
for	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
induction	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
activation	NN	O	O
by	NN	O	O
cAMP	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
two	NN	O	O
parallel	NN	O	O
mitogen-activated	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
pathways	NN	O	O
,	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
signal-related	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
requires	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
a	NN	O	O
co-receptor	NN	O	B-protein
molecule	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
CD28	NN	O	B-protein
,	NN	O	O
that	NN	O	O
activate	NN	O	O
parallel	NN	O	O
and	NN	O	O
partially	NN	O	O
cross-reactive	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

These	NN	O	O
pathways	NN	O	O
are	NN	O	O
disrupted	NN	O	O
by	NN	O	O
agonists	NN	O	O
that	NN	O	O
utilize	NN	O	O
adenylate	NN	O	B-protein
cyclase	NN	O	I-protein
and	NN	O	O
cAMP-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	O
PKA	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
adenylate	NN	O	B-protein
cyclase	NN	O	I-protein
activator	NN	O	O
,	NN	O	O
forskolin	NN	O	O
,	NN	O	O
inhibits	NN	O	O
anti-CD3	NN	O	B-protein
-induced	NN	O	O
shift	NN	O	O
in	NN	O	O
Lck	NN	O	B-protein
electrophoretic	NN	O	O
mobility	NN	O	O
,	NN	O	O
suggesting	NN	O	O
an	NN	O	O
intervention	NN	O	O
at	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
-coupled	NN	O	O
phosphoinositide	NN	O	O
turnover	NN	O	O
that	NN	O	O
precedes	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
.	NN	O	O

The	NN	O	O
shift	NN	O	O
of	NN	O	O
Lck	NN	O	B-protein
following	NN	O	O
direct	NN	O	O
PKC	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
12-O-tetradecanoyl	NN	O	O
phorbol	NN	O	O
13-acetate	NN	O	O
,	NN	O	O
which	NN	O	O
bypasses	NN	O	O
early	NN	O	O
receptor-triggered	NN	O	O
biochemical	NN	O	O
events	NN	O	O
,	NN	O	O
is	NN	O	O
insensitive	NN	O	O
to	NN	O	O
forskolin	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
forskolin	NN	O	O
also	NN	O	O
inhibits	NN	O	O
PKC	NN	O	B-protein
downstream	NN	O	O
events	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
process	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
further	NN	O	O
analyze	NN	O	O
potential	NN	O	O
cross	NN	O	O
points	NN	O	O
between	NN	O	O
positively	NN	O	O
and	NN	O	O
negatively	NN	O	O
regulating	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
activators	NN	O	O
of	NN	O	O
the	NN	O	O
adenylate	NN	O	B-protein
cyclase	NN	O	I-protein
or	NN	O	O
PKA	NN	O	B-protein
on	NN	O	O
two	NN	O	O
parallel	NN	O	O
mitogen-activated	NN	O	O
protein	NN	O	O
kinase	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
mediated	NN	O	O
by	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
ERK	NN	O	B-protein
)	NN	O	O
and	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
a	NN	O	O
PKC	NN	O	B-protein
-specific	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
GF109203X	NN	O	O
,	NN	O	O
or	NN	O	O
PKC-depleted	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
a	NN	O	O
large	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
anti-CD3	NN	O	B-protein
-induced	NN	O	O
ERK	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
PKC	NN	O	B-protein
dependent	NN	O	O
.	NN	O	O

Both	NN	O	O
PKC-	NN	O	O
dependent	NN	O	O
and	NN	O	O
-independent	NN	O	O
activation	NN	O	O
of	NN	O	O
ERK	NN	O	B-protein
were	NN	O	O
sensitive	NN	O	O
to	NN	O	O
inhibition	NN	O	O
by	NN	O	O
forskolin	NN	O	O
or	NN	O	O
a	NN	O	O
cell-permeable	NN	O	O
cAMP	NN	O	O
analogue	NN	O	O
,	NN	O	O
dbcAMP	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
12-O-tetradecanoyl	NN	O	O
phorbol	NN	O	O
13-acetate	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
,	NN	O	O
which	NN	O	O
synergized	NN	O	O
to	NN	O	O
fully	NN	O	O
activate	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
was	NN	O	O
also	NN	O	O
sensitive	NN	O	O
to	NN	O	O
inhibition	NN	O	O
by	NN	O	O
forskolin	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
PKA	NN	O	B-protein
inhibits	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
multiple	NN	O	O
events	NN	O	O
along	NN	O	O
the	NN	O	O
two	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
operating	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
co-receptor	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
and	NN	O	O
posttranscriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
anthracycline-induced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
erythroleukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Aclacinomycin	NN	O	O
(	NN	O	O
ACLA	NN	O	O
)	NN	O	O
and	NN	O	O
doxorubicin	NN	O	O
(	NN	O	O
DOX	NN	O	O
)	NN	O	O
were	NN	O	O
used	NN	O	O
at	NN	O	O
subtoxic	NN	O	O
concentrations	NN	O	O
to	NN	O	O
induce	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
K562	NN	O	I-cell_line
.	NN	O	O

Cell	NN	O	O
hemoglobinization	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
the	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
encoding	NN	O	O
gamma-globin	NN	O	B-protein
and	NN	O	I-protein
porphobilinogen	NN	O	I-protein
deaminase	NN	O	I-protein
(	NN	O	O
PBGD	NN	O	B-protein
)	NN	O	O
,	NN	O	O
an	NN	O	O
enzyme	NN	O	O
of	NN	O	O
heme	NN	O	O
synthesis	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
run-on	NN	O	O
assays	NN	O	O
,	NN	O	O
ACLA	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
induce	NN	O	O
an	NN	O	O
enhancement	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
erythroid	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
gamma-globin	NN	O	B-protein
,	NN	O	O
PBGD	NN	O	B-protein
,	NN	O	O
erythropoietin	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
GATA-1	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
in	NN	O	O
DOX-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
transcription	NN	O	O
rate	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	B-DNA
was	NN	O	O
unchanged	NN	O	O
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
control	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
synthesis	NN	O	O
with	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
indicated	NN	O	O
that	NN	O	O
DOX	NN	O	O
induced	NN	O	O
an	NN	O	O
increased	NN	O	O
stability	NN	O	O
of	NN	O	O
PBGD	NN	O	B-RNA
and	NN	O	I-RNA
GATA-1	NN	O	I-RNA
mRNAs	NN	O	I-RNA
,	NN	O	O
whereas	NN	O	O
ACLA	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
half-lives	NN	O	O
of	NN	O	O
these	NN	O	O
mRNAs	NN	O	B-RNA
.	NN	O	O

Because	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
erythroid	NN	O	B-RNA
mRNA	NN	O	I-RNA
steady-state	NN	O	O
level	NN	O	O
in	NN	O	O
anthracycline-treated	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
cycloheximide	NN	O	O
,	NN	O	O
this	NN	O	O
suggests	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
in	NN	O	O
ACLA-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
mRNA	NN	O	O
stabilization	NN	O	O
in	NN	O	O
DOX-treated	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
dependent	NN	O	O
on	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
GATA-1	NN	O	B-protein
protein	NN	O	O
level	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
increased	NN	O	O
in	NN	O	O
ACLA-treated	NN	O	B-cell_line
but	NN	O	I-cell_line
not	NN	O	I-cell_line
in	NN	O	I-cell_line
DOX-treated	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
two	NN	O	O
anthracyclines	NN	O	O
,	NN	O	O
although	NN	O	O
closely	NN	O	O
related	NN	O	O
in	NN	O	O
their	NN	O	O
structures	NN	O	O
,	NN	O	O
appeared	NN	O	O
to	NN	O	O
act	NN	O	O
as	NN	O	O
differentiation	NN	O	O
inducers	NN	O	O
by	NN	O	O
distinct	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
erythroid	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
transcriptionally	NN	O	O
by	NN	O	O
ACLA	NN	O	O
and	NN	O	O
mainly	NN	O	O
posttranscriptionally	NN	O	O
by	NN	O	O
DOX	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin-7	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
precursor	NN	O	I-cell_line
acute	NN	O	I-cell_line
lymphoblastic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
murine	NN	O	B-cell_line
BAF3	NN	O	I-cell_line
cells	NN	O	I-cell_line
involves	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
mediated	NN	O	O
via	NN	O	O
the	NN	O	O
interleukin-7	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
.	NN	O	O

Interleukin-7	NN	O	B-protein
(	NN	O	O
IL-7	NN	O	B-protein
)	NN	O	O
stimulates	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
and	NN	O	I-cell_type
leukemic	NN	O	I-cell_type
B	NN	O	I-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
precursors	NN	O	I-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
JAK	NN	O	B-protein
/STAT	NN	O	B-protein
pathway	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
IL-7R	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
which	NN	O	O
STAT	NN	O	B-protein
complexes	NN	O	I-protein
are	NN	O	O
formed	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
precursor	NN	O	I-cell_line
acute	NN	O	I-cell_line
lymphoblastic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
(	NN	O	I-cell_line
BCP-ALL	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
IL-7	NN	O	B-protein
.	NN	O	O

Gel	NN	O	O
retardation	NN	O	O
assays	NN	O	O
with	NN	O	O
STAT-binding	NN	O	O
oligonucleotides	NN	O	O
showed	NN	O	O
that	NN	O	O
IL-7	NN	O	B-protein
induces	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
two	NN	O	O
major	NN	O	O
STAT	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
BCP-ALL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Supershifts	NN	O	O
with	NN	O	O
anti-STAT	NN	O	O
antibodies	NN	O	O
identified	NN	O	O
these	NN	O	O
as	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

This	NN	O	O
pattern	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
seen	NN	O	O
in	NN	O	O
all	NN	O	O
BCP-ALL	NN	O	O
cases	NN	O	O
that	NN	O	O
respond	NN	O	O
to	NN	O	O
IL-7	NN	O	B-protein
in	NN	O	O
proliferation	NN	O	O
assays	NN	O	O
.	NN	O	O

IL-7	NN	O	B-protein
also	NN	O	O
induced	NN	O	O
STAT	NN	O	B-protein
/DNA	NN	O	O
binding	NN	O	O
in	NN	O	O
BCP-ALL	NN	O	O
cases	NN	O	O
that	NN	O	O
failed	NN	O	O
to	NN	O	O
proliferate	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-7	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
IL-7R	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
the	NN	O	O
JAK	NN	O	B-protein
/STAT	NN	O	B-protein
pathway	NN	O	O
per	NN	O	O
se	NN	O	O
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
for	NN	O	O
proliferation	NN	O	O
induction	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
IL-7	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
IL-7R	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
transfectants	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
pro-B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
BAF3	NN	O	I-cell_line
were	NN	O	O
made	NN	O	O
that	NN	O	O
express	NN	O	O
chimeric	NN	O	B-protein
receptors	NN	O	I-protein
consisting	NN	O	O
of	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
human	NN	O	B-protein
granulocyte	NN	O	I-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
G-CSF-R	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
transmembrane	NN	O	B-protein
and	NN	O	I-protein
intracellular	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
human	NN	O	B-protein
IL-7R	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
G-CSF-R/IL-7R	NN	O	I-protein
alpha	NN	O	I-protein
with	NN	O	O
G-CSF	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
full	NN	O	O
proliferative	NN	O	O
response	NN	O	O
and	NN	O	O
induced	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
JAK1	NN	O	B-protein
but	NN	O	O
not	NN	O	O
JAK2	NN	O	B-protein
.	NN	O	O

Major	NN	O	O
STAT	NN	O	B-protein
complexes	NN	O	I-protein
activated	NN	O	O
by	NN	O	O
G-CSF-R/IL-7R	NN	O	B-protein
alpha	NN	O	I-protein
contained	NN	O	O
STAT1	NN	O	B-protein
or	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
while	NN	O	O
some	NN	O	O
formation	NN	O	O
of	NN	O	O
STAT3-containing	NN	O	B-protein
complexes	NN	O	I-protein
was	NN	O	O
also	NN	O	O
seen	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
establish	NN	O	O
that	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
and	NN	O	O
possibly	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
are	NN	O	O
activated	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
of	NN	O	O
precursor	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
IL-7	NN	O	B-protein
.	NN	O	O

The	NN	O	O
data	NN	O	O
further	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
IL-7R	NN	O	B-protein
alpha	NN	O	I-protein
chains	NN	O	I-protein
are	NN	O	O
directly	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
JAKs	NN	O	B-protein
and	NN	O	O
STATs	NN	O	B-protein
and	NN	O	O
have	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
proliferative	NN	O	O
signaling	NN	O	O
in	NN	O	O
precursor	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Stimulation	NN	O	O
of	NN	O	O
human	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
and	NN	O	O
CD40	NN	O	B-protein
antigen	NN	O	O
expression	NN	O	O
by	NN	O	O
phosphorothioate	NN	O	O
oligonucleotides	NN	O	O
complementary	NN	O	O
to	NN	O	O
hepatitis	NN	O	B-DNA
B	NN	O	I-DNA
virus	NN	O	I-DNA
genome	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and	NN	O	O
CD40	NN	O	B-protein
antigen	NN	O	O
expression	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
cytotoxicity	NN	O	O
to	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
of	NN	O	O
antisense	NN	O	O
phosphorothioate	NN	O	O
oligodeoxynucleotides	NN	O	O
complementary	NN	O	O
to	NN	O	O
the	NN	O	O
SP	NN	O	B-DNA
II	NN	O	I-DNA
promoter	NN	O	I-DNA
of	NN	O	O
HBV	NN	O	B-RNA
mRNA	NN	O	I-RNA
(	NN	O	O
sequence	NN	O	O
I	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
gene	NN	O	I-DNA
(	NN	O	O
sequence	NN	O	O
II	NN	O	O
)	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
.	NN	O	O

The	NN	O	O
oligo	NN	O	O
sequence	NN	O	O
I	NN	O	O
stimulated	NN	O	O
proliferation	NN	O	O
of	NN	O	O
both	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
,	NN	O	I-cell_type
to	NN	O	I-cell_type
a	NN	O	I-cell_type
lesser	NN	O	I-cell_type
extent	NN	O	I-cell_type
,	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
percentage	NN	O	O
of	NN	O	O
cells	NN	O	O
expressing	NN	O	O
CD40	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_line
and	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
co-cultures	NN	O	I-cell_line
increased	NN	O	O
from	NN	O	O
4.2	NN	O	O
%	NN	O	O
to	NN	O	O
13.8	NN	O	O
%	NN	O	O
after	NN	O	O
oligo	NN	O	O
stimulation	NN	O	O
in	NN	O	O
patients	NN	O	O
,	NN	O	O
while	NN	O	O
it	NN	O	O
increased	NN	O	O
form	NN	O	O
4.7	NN	O	O
%	NN	O	O
to	NN	O	O
48.6	NN	O	O
%	NN	O	O
in	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

The	NN	O	O
sense	NN	O	O
sequence	NN	O	O
(	NN	O	O
sequence	NN	O	O
III	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
gene	NN	O	I-DNA
also	NN	O	O
enhanced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
antigen	NN	O	I-protein
in	NN	O	O
patients	NN	O	O
with	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
.	NN	O	O

The	NN	O	O
proportion	NN	O	O
of	NN	O	O
CD40	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
26	NN	O	O
%	NN	O	O
)	NN	O	O
in	NN	O	O
a	NN	O	O
resting	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
preparation	NN	O	I-cell_line
from	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
patients	NN	O	O
decreased	NN	O	O
to	NN	O	O
zero	NN	O	O
after	NN	O	O
a	NN	O	O
5-day	NN	O	O
culture	NN	O	O
with	NN	O	O
sequence	NN	O	O
I	NN	O	O
,	NN	O	O
but	NN	O	O
IgG	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
culture	NN	O	O
supernatant	NN	O	O
increased	NN	O	O
.	NN	O	O

The	NN	O	O
cytotoxic	NN	O	O
properties	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
were	NN	O	O
not	NN	O	O
influenced	NN	O	O
by	NN	O	O
the	NN	O	O
oligos	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
antisense	NN	O	O
oligos	NN	O	O
against	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
virus	NN	O	O
(	NN	O	O
HBV	NN	O	O
)	NN	O	O
have	NN	O	O
mitogenic	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
a	NN	O	O
non-specific	NN	O	O
manner	NN	O	O
and	NN	O	O
may	NN	O	O
activate	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
express	NN	O	O
CD40	NN	O	B-protein
antigen	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	B-protein
protein	NN	O	I-protein
1	NN	O	I-protein
-dependent	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
Ca2+/calmodulin	NN	O	B-protein
kinase	NN	O	I-protein
type	NN	O	I-protein
IV/Gr	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Ca2+/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
CaMK	NN	O	I-protein
)	NN	O	I-protein
type	NN	O	I-protein
IV/Gr	NN	O	I-protein
is	NN	O	O
selectively	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
activated	NN	O	O
after	NN	O	O
signaling	NN	O	O
via	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
it	NN	O	O
mediates	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
transcriptional	NN	O	O
events	NN	O	O
that	NN	O	O
follow	NN	O	O
TCR	NN	O	B-protein
engagement	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
CaMKIV/Gr	NN	O	B-protein
induces	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
activation	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
alone	NN	O	O
or	NN	O	O
in	NN	O	O
synergy	NN	O	O
with	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
mitogens	NN	O	I-protein
and	NN	O	O
with	NN	O	O
the	NN	O	O
p21ras	NN	O	B-protein
oncoprotein	NN	O	I-protein
.	NN	O	O

CaMKIV/	NN	O	B-protein
Gr	NN	O	I-protein
signaling	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
but	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
p21ras	NN	O	B-DNA
or	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

AP-1	NN	O	B-protein
is	NN	O	O
an	NN	O	O
integral	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	I-protein
NFAT	NN	O	I-protein
)	NN	O	I-protein
transcriptional	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
CaMKIV/Gr	NN	O	B-protein
reconstitutes	NN	O	O
the	NN	O	O
capacity	NN	O	O
of	NN	O	O
the	NN	O	O
cytosolic	NN	O	O
component	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
to	NN	O	O
direct	NN	O	O
transcription	NN	O	O
from	NN	O	O
NFAT	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
non-	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
reveal	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
for	NN	O	O
CaMKIV/Gr	NN	O	B-protein
as	NN	O	O
a	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-regulated	NN	O	O
activator	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

IL-12	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
immediate-early	NN	O	B-DNA
activation	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

The	NN	O	O
responses	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
to	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
involving	NN	O	O
proliferation	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
,	NN	O	O
are	NN	O	O
only	NN	O	O
partially	NN	O	O
overlapping	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
depend	NN	O	O
on	NN	O	O
induced	NN	O	O
differential	NN	O	O
expression	NN	O	O
of	NN	O	O
specific	NN	O	O
sets	NN	O	O
of	NN	O	O
genes	NN	O	B-protein
.	NN	O	O

Using	NN	O	O
reverse-transcription	NN	O	O
PCR	NN	O	O
differential	NN	O	O
display	NN	O	O
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
an	NN	O	O
mRNA	NN	O	B-RNA
species	NN	O	I-RNA
expressed	NN	O	O
in	NN	O	O
IL-2	NN	O	B-cell_type
-but	NN	O	I-cell_type
not	NN	O	I-cell_type
IL-12-stimulated	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
encoding	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
egr-1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
expressed	NN	O	O
with	NN	O	O
fast	NN	O	O
kinetics	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
upon	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
mRNA-encoding	NN	O	B-protein
members	NN	O	I-protein
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
demonstrated	NN	O	O
that	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
junB	NN	O	B-DNA
are	NN	O	O
also	NN	O	O
expressed	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
expression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
junD	NN	O	B-DNA
is	NN	O	O
not	NN	O	O
modified	NN	O	O
by	NN	O	O
either	NN	O	O
cytokine	NN	O	B-protein
.	NN	O	O

Accordingly	NN	O	O
,	NN	O	O
increased	NN	O	O
AP-1	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
-dependent	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
were	NN	O	O
detected	NN	O	O
exclusively	NN	O	O
in	NN	O	O
IL-2-stimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	B-protein
reported	NN	O	O
to	NN	O	O
regulate	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
both	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
induce	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
in	NN	O	O
NK	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
only	NN	O	O
IL-2	NN	O	B-protein
induces	NN	O	O
bcl-2	NN	O	B-DNA
expression	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
provide	NN	O	O
the	NN	O	O
first	NN	O	O
demonstration	NN	O	O
that	NN	O	O
IL-12	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
does	NN	O	O
not	NN	O	O
involve	NN	O	O
expression	NN	O	O
of	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
immediate-early	NN	O	B-DNA
activation	NN	O	I-DNA
genes	NN	O	I-DNA
family	NN	O	I-DNA
(	NN	O	O
egr-1	NN	O	B-DNA
,	NN	O	O
c-fos	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
junB	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
AP-1	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
,	NN	O	O
or	NN	O	O
bcl-2	NN	O	B-DNA
expression	NN	O	O
.	NN	O	O

This	NN	O	O
indicates	NN	O	O
that	NN	O	O
functional	NN	O	O
differences	NN	O	O
observed	NN	O	O
in	NN	O	O
IL-2-	NN	O	B-cell_line
and	NN	O	I-cell_line
IL-12-stimulated	NN	O	I-cell_line
cells	NN	O	I-cell_line
may	NN	O	O
depend	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
on	NN	O	O
differential	NN	O	O
gene	NN	O	O
regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Active	NN	O	O
suppression	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
transactivator-encoding	NN	O	I-DNA
AIR-1	NN	O	I-DNA
locus	NN	O	I-DNA
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
observed	NN	O	O
during	NN	O	O
differentiation	NN	O	O
from	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
the	NN	O	O
genetic	NN	O	O
control	NN	O	O
of	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
(	NN	O	I-DNA
MHC	NN	O	I-DNA
)	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
during	NN	O	O
the	NN	O	O
transition	NN	O	O
from	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
to	NN	O	O
plasma	NN	O	B-cell_type
cell	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
analyzed	NN	O	O
.	NN	O	O

Class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
are	NN	O	O
not	NN	O	O
expressed	NN	O	O
in	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
because	NN	O	O
of	NN	O	O
an	NN	O	O
active	NN	O	O
suppression	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
abrogation	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
plasma	NN	O	O
cell-specific	NN	O	O
repressor	NN	O	O
function	NN	O	O
,	NN	O	O
designated	NN	O	O
SIR	NN	O	B-DNA
(	NN	O	O
suppressor	NN	O	B-DNA
of	NN	O	I-DNA
immune	NN	O	I-DNA
response	NN	O	I-DNA
genes	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
does	NN	O	O
not	NN	O	O
act	NN	O	O
directly	NN	O	O
on	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
instead	NN	O	O
on	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
AIR-1	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
whose	NN	O	O
product	NN	O	O
,	NN	O	O
the	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
is	NN	O	O
fundamental	NN	O	O
for	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
was	NN	O	O
unambiguously	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
plasmacytoma	NN	O	B-cell_line
x	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
hybrids	NN	O	I-cell_line
carrying	NN	O	O
an	NN	O	O
AIR-1	NN	O	B-DNA
locus	NN	O	I-DNA
derived	NN	O	O
from	NN	O	O
CIITA-expressing	NN	O	B-cell_line
cells	NN	O	I-cell_line
do	NN	O	O
not	NN	O	O
express	NN	O	O
CIITA-specific	NN	O	B-RNA
transcripts	NN	O	I-RNA
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	O
containing	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CIITA	NN	O	I-DNA
coding	NN	O	I-DNA
sequence	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
an	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
restores	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
MHC	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
hybrids	NN	O	O
and	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
de	NN	O	O
novo	NN	O	O
expression	NN	O	O
of	NN	O	O
mouse	NN	O	B-DNA
MHC	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
both	NN	O	O
the	NN	O	O
mouse	NN	O	B-cell_line
plasmacytoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
hybrids	NN	O	B-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
confirm	NN	O	O
and	NN	O	O
extend	NN	O	O
the	NN	O	O
notion	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
conservation	NN	O	O
of	NN	O	O
the	NN	O	O
AIR-1	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
across	NN	O	O
species	NN	O	O
barriers	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
in	NN	O	O
CIITA-transfected	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybrids	NN	O	I-cell_line
,	NN	O	O
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
HLA-DQ	NN	O	I-protein
heterodimer	NN	O	I-protein
was	NN	O	O
not	NN	O	O
observed	NN	O	O
.	NN	O	O

This	NN	O	O
result	NN	O	O
was	NN	O	O
not	NN	O	O
attributable	NN	O	O
to	NN	O	O
lack	NN	O	O
of	NN	O	O
HLA-DQ	NN	O	O
alpha	NN	O	O
or	NN	O	O
-DQ	NN	O	O
beta	NN	O	O
transcription	NN	O	O
,	NN	O	O
because	NN	O	O
both	NN	O	O
transcripts	NN	O	O
were	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
CIITA-transfected	NN	O	B-cell_line
hybrids	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
at	NN	O	O
reduced	NN	O	O
levels	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
further	NN	O	O
support	NN	O	O
our	NN	O	O
previous	NN	O	O
observations	NN	O	O
on	NN	O	O
the	NN	O	O
distinct	NN	O	O
regulation	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
HLA-DQ	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
subset	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
thus	NN	O	O
controlled	NN	O	O
at	NN	O	O
the	NN	O	O
posttranscriptional	NN	O	O
level	NN	O	O
by	NN	O	O
a	NN	O	O
CIITA	NN	O	B-protein
-independent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

-DOCSTART-	O

Requirements	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D-mediated	NN	O	O
gene	NN	O	O
regulation	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Mature	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
are	NN	O	O
unique	NN	O	O
targets	NN	O	O
of	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
)	NN	O	O
in	NN	O	O
that	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
are	NN	O	O
not	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
,	NN	O	O
and	NN	O	O
specific	NN	O	O
cellular	NN	O	O
activation	NN	O	O
signals	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
both	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
and	NN	O	O
establishment	NN	O	O
of	NN	O	O
reactivity	NN	O	O
to	NN	O	O
the	NN	O	O
lipophilic	NN	O	O
ligand	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
IL-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
prior	NN	O	O
activation	NN	O	O
,	NN	O	O
induces	NN	O	O
a	NN	O	O
weak	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
expression	NN	O	O
but	NN	O	O
fails	NN	O	O
to	NN	O	O
generate	NN	O	O
vitamin	NN	O	B-protein
D-responsive	NN	O	I-protein
element	NN	O	I-protein
(	NN	O	I-protein
VDRE	NN	O	I-protein
)	NN	O	I-protein
-reactive	NN	O	I-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
complexes	NN	O	I-protein
or	NN	O	O
to	NN	O	O
initiate	NN	O	O
the	NN	O	O
genomic	NN	O	O
transcription	NN	O	O
of	NN	O	O
25-hydroxyvitamin	NN	O	B-protein
D3	NN	O	I-protein
24-hydroxylase	NN	O	I-protein
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
either	NN	O	O
ligation	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
Ag	NN	O	I-protein
or	NN	O	O
cross-linking	NN	O	O
the	NN	O	O
Ig	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
also	NN	O	O
insufficient	NN	O	O
to	NN	O	O
render	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
responsive	NN	O	O
to	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
apparent	NN	O	O
lack	NN	O	O
of	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
secosterol	NN	O	O
can	NN	O	O
be	NN	O	O
overcome	NN	O	O
by	NN	O	O
stimulation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
these	NN	O	O
cellular	NN	O	O
activation	NN	O	O
signals	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
sufficient	NN	O	O
to	NN	O	O
lead	NN	O	O
to	NN	O	O
G1	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
,	NN	O	O
cellular	NN	O	O
activation	NN	O	O
associated	NN	O	O
with	NN	O	O
stimulation	NN	O	O
of	NN	O	O
such	NN	O	O
a	NN	O	O
progression	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
sufficient	NN	O	O
for	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
message	NN	O	O
and	NN	O	O
protein	NN	O	O
and	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
establishment	NN	O	O
of	NN	O	O
VDRE	NN	O	B-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
24-hydroxylase	NN	O	B-protein
message	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
biologic	NN	O	O
functions	NN	O	O
are	NN	O	O
modulated	NN	O	O
,	NN	O	O
in	NN	O	O
that	NN	O	O
the	NN	O	O
hormone	NN	O	O
inhibits	NN	O	O
proliferation	NN	O	O
in	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
the	NN	O	O
activated	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
reactivity	NN	O	O
to	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
is	NN	O	O
tightly	NN	O	O
regulated	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
requiring	NN	O	O
specific	NN	O	O
signals	NN	O	O
for	NN	O	O
its	NN	O	O
initiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cross	NN	O	O
talk	NN	O	O
between	NN	O	O
cell	NN	O	O
death	NN	O	O
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
:	NN	O	O
BCL-2	NN	O	B-protein
regulates	NN	O	O
NFAT	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
.	NN	O	O

BCL-2-deficient	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
demonstrate	NN	O	O
accelerated	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
and	NN	O	O
increased	NN	O	O
apoptosis	NN	O	O
following	NN	O	O
activation	NN	O	O
.	NN	O	O

Increasing	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
BCL-2	NN	O	B-protein
retarded	NN	O	O
the	NN	O	O
G0	NN	O	O
--	NN	O	O
>	NN	O	O
S	NN	O	O
transition	NN	O	O
,	NN	O	O
sustained	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
cyclin-dependent	NN	O	B-protein
kinase	NN	O	I-protein
inhibitor	NN	O	O
p27Kip1	NN	O	O
,	NN	O	O
and	NN	O	O
repressed	NN	O	O
postactivation	NN	O	O
death	NN	O	O
.	NN	O	O

Proximal	NN	O	O
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
and	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
were	NN	O	O
unaffected	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
transcription	NN	O	O
and	NN	O	O
synthesis	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
and	NN	O	O
other	NN	O	O
delayed	NN	O	O
early	NN	O	O
cytokines	NN	O	B-protein
were	NN	O	O
markedly	NN	O	O
attenuated	NN	O	O
by	NN	O	O
BCL-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
cysteine	NN	O	O
protease	NN	O	O
inhibitor	NN	O	O
that	NN	O	O
also	NN	O	O
blocks	NN	O	O
apoptosis	NN	O	O
had	NN	O	O
no	NN	O	O
substantial	NN	O	O
affect	NN	O	O
upon	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
.	NN	O	O

InterleUkin	NN	O	B-protein
2	NN	O	I-protein
expression	NN	O	O
requires	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
which	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
(	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
)	NN	O	O
and	NN	O	O
NFAT	NN	O	B-protein
-mediated	NN	O	O
transactivation	NN	O	O
were	NN	O	O
impaired	NN	O	O
by	NN	O	O
BCL-2	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
select	NN	O	O
genetic	NN	O	O
aberrations	NN	O	O
in	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
pathway	NN	O	O
reveal	NN	O	O
a	NN	O	O
cell	NN	O	O
autonomous	NN	O	O
coregulation	NN	O	O
of	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interaction	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
lymphotropic	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
tax	NN	O	I-protein
transactivator	NN	O	I-protein
with	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
IIA	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
lymphotropic	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
40-kDa	NN	O	B-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
which	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
HTLV-1	NN	O	O
gene	NN	O	O
regulation	NN	O	O
and	NN	O	O
virus-induced	NN	O	O
cellular	NN	O	O
transformation	NN	O	O
.	NN	O	O

Tax	NN	O	B-protein
is	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
DNA	NN	O	O
through	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
site-specific	NN	O	B-protein
activators	NN	O	I-protein
cyclic	NN	O	B-protein
AMP-responsive	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
serum	NN	O	B-protein
response	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
to	NN	O	O
the	NN	O	O
DNA	NN	O	O
positions	NN	O	O
Tax	NN	O	B-protein
for	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
basal	NN	O	O
transcriptional	NN	O	O
machinery	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
several	NN	O	O
independent	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
now	NN	O	O
report	NN	O	O
a	NN	O	O
physical	NN	O	O
and	NN	O	O
functional	NN	O	O
interaction	NN	O	O
between	NN	O	O
Tax	NN	O	B-protein
and	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
TFIIA	NN	O	B-protein
.	NN	O	O

First	NN	O	O
,	NN	O	O
Tax	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
35-kDa	NN	O	B-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
of	NN	O	O
TFIIA	NN	O	B-protein
in	NN	O	O
the	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
interaction	NN	O	O
system	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
two	NN	O	O
previously	NN	O	O
characterized	NN	O	O
mutants	NN	O	B-protein
with	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
Tax	NN	O	B-protein
,	NN	O	O
M32	NN	O	B-protein
(	NN	O	O
Y196A	NN	O	O
,	NN	O	O
K197S	NN	O	O
)	NN	O	O
and	NN	O	O
M41	NN	O	B-protein
(	NN	O	O
H287A	NN	O	O
,	NN	O	O
P288S	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
defective	NN	O	O
in	NN	O	O
Tax	NN	O	B-protein
-activated	NN	O	O
transcription	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
TFIIA	NN	O	B-protein
in	NN	O	O
this	NN	O	O
assay	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
a	NN	O	O
glutathione-S-transferase	NN	O	O
(	NN	O	O
GST	NN	O	B-protein
)	NN	O	O
affinity-binding	NN	O	O
assay	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
holo-	NN	O	O
TFIIA	NN	O	B-protein
with	NN	O	O
GST-Tax	NN	O	B-protein
was	NN	O	O
20-fold	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
observed	NN	O	O
with	NN	O	O
either	NN	O	O
the	NN	O	O
GST-Tax	NN	O	B-protein
M32	NN	O	I-protein
activation	NN	O	I-protein
mutant	NN	O	I-protein
or	NN	O	O
the	NN	O	O
GST	NN	O	O
control	NN	O	O
.	NN	O	O

Third	NN	O	O
,	NN	O	O
a	NN	O	O
coimmunoprecipitation	NN	O	O
assay	NN	O	O
showed	NN	O	O
that	NN	O	O
in	NN	O	O
HTLV-1-infected	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
,	NN	O	O
Tax	NN	O	B-protein
and	NN	O	O
TFIIA	NN	O	B-protein
were	NN	O	O
associated	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
TFIIA	NN	O	B-protein
facilitates	NN	O	O
Tax	NN	O	B-protein
transactivation	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
studies	NN	O	O
showed	NN	O	O
reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
-activated	NN	O	O
transcription	NN	O	O
in	NN	O	O
cell	NN	O	O
extracts	NN	O	O
depleted	NN	O	O
of	NN	O	O
TFIIA	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
transfection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
TFIIA	NN	O	B-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
enhanced	NN	O	O
Tax	NN	O	B-protein
-activated	NN	O	O
transcription	NN	O	O
of	NN	O	O
an	NN	O	O
HTLV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat-chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
study	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
with	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
TFIIA	NN	O	B-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
Tax	NN	O	B-protein
-mediated	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Stat1	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
the	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
results	NN	O	O
in	NN	O	O
a	NN	O	O
cascade	NN	O	O
of	NN	O	O
gene	NN	O	O
activation	NN	O	O
and	NN	O	O
subsequent	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
into	NN	O	O
effector	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
belonging	NN	O	O
to	NN	O	O
the	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
was	NN	O	O
analyzed	NN	O	O
in	NN	O	O
PHA-activated	NN	O	B-cell_line
mononuclear	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
purified	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
activated	NN	O	O
by	NN	O	O
cross-linking	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
CD3	NN	O	I-protein
.	NN	O	O

Cell	NN	O	O
activation	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
delayed	NN	O	O
induction	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
sustained	NN	O	O
for	NN	O	O
several	NN	O	O
days	NN	O	O
,	NN	O	O
was	NN	O	O
composed	NN	O	O
predominantly	NN	O	O
of	NN	O	O
Stat1	NN	O	B-protein
and	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
cycloheximide	NN	O	O
and	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
.	NN	O	O

Increased	NN	O	O
Stat1	NN	O	O
and	NN	O	O
Stat3	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
levels	NN	O	O
were	NN	O	O
detected	NN	O	O
,	NN	O	O
respectively	NN	O	O
4	NN	O	O
and	NN	O	O
24	NN	O	O
h	NN	O	O
after	NN	O	O
activation	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
cAMP	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
,	NN	O	O
which	NN	O	O
skews	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
toward	NN	O	O
a	NN	O	O
Th2	NN	O	O
pattern	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
preferential	NN	O	O
suppression	NN	O	O
of	NN	O	O
Stat1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

cAMP	NN	O	O
inhibited	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
components	NN	O	I-protein
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
inhibit	NN	O	O
IL-4	NN	O	B-protein
receptor	NN	O	I-protein
alpha-chain	NN	O	I-protein
and	NN	O	O
CD69	NN	O	B-protein
expression	NN	O	O
or	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
activator	NN	O	B-protein
protein	NN	O	I-protein
1	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

cAMP	NN	O	O
signaling	NN	O	O
inhibited	NN	O	O
Stat1	NN	O	B-protein
at	NN	O	O
several	NN	O	O
different	NN	O	O
levels	NN	O	O
,	NN	O	O
including	NN	O	O
suppression	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
Stat1	NN	O	O
protein	NN	O	O
and	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
a	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
that	NN	O	O
regulates	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
functional	NN	O	O
phenotype	NN	O	O
and	NN	O	O
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
-activated	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
-STAT	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
by	NN	O	O
antisense	NN	O	O
oligonucleotides	NN	O	O
inhibits	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
but	NN	O	O
not	NN	O	O
respiratory	NN	O	O
burst	NN	O	O
during	NN	O	O
phorbol	NN	O	O
ester-induced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
human	NN	O	I-cell_line
monoblastic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
studied	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
gene	NN	O	I-DNA
c-jun	NN	O	B-DNA
in	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
-induced	NN	O	O
differentiation	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
human	NN	O	I-cell_line
monoblastic	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
using	NN	O	O
c-jun	NN	O	B-DNA
-specific	NN	O	O
antisense	NN	O	O
(	NN	O	O
AS	NN	O	O
)	NN	O	O
phosphorothioate	NN	O	O
oligonucleotides	NN	O	O
.	NN	O	O

In	NN	O	O
selecting	NN	O	O
the	NN	O	O
most	NN	O	O
specific	NN	O	O
and	NN	O	O
potent	NN	O	O
oligonucleotide	NN	O	O
sequence	NN	O	O
,	NN	O	O
we	NN	O	O
performed	NN	O	O
extensive	NN	O	O
analyses	NN	O	O
for	NN	O	O
the	NN	O	O
binding	NN	O	O
specificity	NN	O	O
between	NN	O	O
all	NN	O	O
candidates	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
AS	NN	O	O
oligonucleotides	NN	O	O
and	NN	O	O
the	NN	O	O
whole	NN	O	O
sequences	NN	O	O
in	NN	O	O
GenBank	NN	O	O
database	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
computer	NN	O	O
program	NN	O	O
.	NN	O	O

Among	NN	O	O
the	NN	O	O
20	NN	O	O
selected	NN	O	O
oligonucleotides	NN	O	O
,	NN	O	O
two	NN	O	O
potent	NN	O	O
15-mer	NN	O	O
AS	NN	O	O
oligonucleotides	NN	O	O
(	NN	O	O
C-JUN	NN	O	O
AS	NN	O	O
oligonucleotides	NN	O	O
)	NN	O	O
exhibited	NN	O	O
significant	NN	O	O
inhibition	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
between	NN	O	O
2	NN	O	O
and	NN	O	O
10	NN	O	O
microM	NN	O	O
.	NN	O	O

Reverse	NN	O	O
transcription-PCR	NN	O	O
and	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
10	NN	O	O
microM	NN	O	O
of	NN	O	O
C-JUN	NN	O	O
AS	NN	O	O
oligonucleotides	NN	O	O
reduced	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
at	NN	O	O
both	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
and	NN	O	O
protein	NN	O	O
levels	NN	O	O
.	NN	O	O

More	NN	O	O
importantly	NN	O	O
,	NN	O	O
C-JUN	NN	O	O
AS	NN	O	O
oligonucleotides	NN	O	O
showed	NN	O	O
distinct	NN	O	O
effects	NN	O	O
on	NN	O	O
two	NN	O	O
markers	NN	O	O
of	NN	O	O
PMA-induced	NN	O	O
differentiation	NN	O	O
;	NN	O	O
the	NN	O	O
C-JUN	NN	O	O
AS	NN	O	O
oligonucleotides	NN	O	O
inhibited	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
,	NN	O	O
whereas	NN	O	O
they	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
another	NN	O	O
marker	NN	O	O
of	NN	O	O
differentiation	NN	O	O
,	NN	O	O
respiratory	NN	O	O
burst	NN	O	O
(	NN	O	O
measured	NN	O	O
by	NN	O	O
nitro	NN	O	O
blue	NN	O	O
tetrazolium	NN	O	O
reduction	NN	O	O
assay	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
in	NN	O	O
both	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
PMA-induced	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
PMA-induced	NN	O	O
respiratory	NN	O	O
burst	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
on	NN	O	O
LPS-induced	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NF-IL6	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
which	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
IL-6	NN	O	B-protein
regulation	NN	O	O
.	NN	O	O

Interleukin-10	NN	O	B-protein
(	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
,	NN	O	O
like	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
inhibit	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
showed	NN	O	O
that	NN	O	O
both	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
inhibit	NN	O	O
LPS-induced	NN	O	B-RNA
IL-6	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
transcription	NN	O	O
rate	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
strong	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-protein
transcription	NN	O	O
rate	NN	O	O
prompted	NN	O	O
us	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
questioned	NN	O	O
whether	NN	O	O
or	NN	O	O
not	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
affected	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-protein
transcription	NN	O	O
rate	NN	O	O
,	NN	O	O
namely	NN	O	O
activator	NN	O	O
protein-1	NN	O	O
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
IL-6	NN	O	B-protein
(	NN	O	O
NF-IL6	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSAs	NN	O	O
)	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
inhibited	NN	O	O
LPS-induced	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
inhibiting	NN	O	O
effect	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
was	NN	O	O
slightly	NN	O	O
more	NN	O	O
pronounced	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

Downregulation	NN	O	O
of	NN	O	O
LPS-induced	NN	O	B-protein
AP-1	NN	O	I-protein
was	NN	O	O
accompanied	NN	O	O
,	NN	O	O
and	NN	O	O
thus	NN	O	O
possibly	NN	O	O
explained	NN	O	O
,	NN	O	O
by	NN	O	O
a	NN	O	O
reduced	NN	O	O
expression	NN	O	O
at	NN	O	O
mRNA	NN	O	B-RNA
level	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
major	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
namely	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	B-DNA
as	NN	O	O
determined	NN	O	O
by	NN	O	O
Northern	NN	O	O
experiments	NN	O	O
.	NN	O	O

Binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-IL6	NN	O	B-protein
was	NN	O	O
also	NN	O	O
strongly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
whereas	NN	O	O
IL-10	NN	O	B-protein
showed	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

NF-IL6	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
were	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
or	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
affects	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
preexisting	NN	O	O
NF-IL6	NN	O	B-protein
.	NN	O	O

Neither	NN	O	O
IL-10	NN	O	B-protein
nor	NN	O	O
IL-4	NN	O	B-protein
inhibited	NN	O	O
LPS-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
agreement	NN	O	O
with	NN	O	O
this	NN	O	O
finding	NN	O	O
,	NN	O	O
Northern	NN	O	O
experiments	NN	O	O
where	NN	O	O
p65	NN	O	O
and	NN	O	O
p105	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
were	NN	O	O
determined	NN	O	O
,	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	O
factor	NN	O	O
were	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
or	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
neither	NN	O	O
IL-10	NN	O	B-protein
nor	NN	O	O
IL-4	NN	O	B-protein
showed	NN	O	O
any	NN	O	O
effect	NN	O	O
on	NN	O	O
I-kappa	NN	O	B-RNA
B	NN	O	I-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
Northern	NN	O	O
experiments	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
similarly	NN	O	O
affect	NN	O	O
IL-6	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
for	NN	O	O
IL-4	NN	O	B-protein
this	NN	O	O
was	NN	O	O
accompanied	NN	O	O
with	NN	O	O
a	NN	O	O
reduction	NN	O	O
of	NN	O	O
AP-1	NN	O	O
and	NN	O	O
NF-IL6	NN	O	O
binding	NN	O	O
activity	NN	O	O
whereas	NN	O	O
IL-10	NN	O	B-protein
only	NN	O	O
inhibited	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
by	NN	O	O
core-binding	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
known	NN	O	O
to	NN	O	O
involve	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Sp1	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	I-DNA
enhancer	NN	O	I-DNA
also	NN	O	O
encompass	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
core-binding	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
CBF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Significantly	NN	O	O
,	NN	O	O
the	NN	O	O
CBF	NN	O	B-DNA
sites	NN	O	I-DNA
are	NN	O	O
in	NN	O	O
each	NN	O	O
case	NN	O	O
contained	NN	O	O
within	NN	O	O
the	NN	O	O
minimum	NN	O	O
essential	NN	O	B-DNA
core	NN	O	I-DNA
regions	NN	O	I-DNA
required	NN	O	O
for	NN	O	O
inducible	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
these	NN	O	O
core	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	O
and	NN	O	O
promoter	NN	O	O
each	NN	O	O
encompass	NN	O	O
closely	NN	O	B-DNA
linked	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
CBF	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NFATp	NN	O	B-protein
.	NN	O	O

The	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
CBF	NN	O	I-DNA
site	NN	O	I-DNA
TGTGGTCA	NN	O	O
is	NN	O	O
located	NN	O	O
51	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
also	NN	O	O
overlaps	NN	O	O
a	NN	O	O
YY-1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
2-bp	NN	O	B-DNA
mutation	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
CBF	NN	O	B-protein
site	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
2-3-fold	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
both	NN	O	O
a	NN	O	O
69-bp	NN	O	B-DNA
proximal	NN	O	I-DNA
promoter	NN	O	I-DNA
fragment	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
627-bp	NN	O	B-DNA
full-length	NN	O	I-DNA
promoter	NN	O	I-DNA
fragment	NN	O	I-DNA
.	NN	O	O

Stepwise	NN	O	O
deletions	NN	O	O
into	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
also	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
CBF	NN	O	B-protein
site	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
YY-1	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
efficient	NN	O	O
induction	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
AML1	NN	O	B-DNA
and	NN	O	I-DNA
CBF	NN	O	I-DNA
beta	NN	O	I-DNA
genes	NN	O	I-DNA
that	NN	O	O
encode	NN	O	O
CBF	NN	O	B-protein
each	NN	O	O
have	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
influence	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
and	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
as	NN	O	O
proto-oncogenes	NN	O	B-DNA
in	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
adds	NN	O	O
GM-CSF	NN	O	B-protein
to	NN	O	O
a	NN	O	O
growing	NN	O	O
list	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
receptors	NN	O	O
that	NN	O	O
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
CBF	NN	O	B-protein
and	NN	O	O
which	NN	O	O
control	NN	O	O
the	NN	O	O
growth	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
hemopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
GM-CSF	NN	O	B-DNA
locus	NN	O	I-DNA
may	NN	O	O
represent	NN	O	O
one	NN	O	O
of	NN	O	O
several	NN	O	O
target	NN	O	O
genes	NN	O	B-protein
that	NN	O	O
are	NN	O	O
dysregulated	NN	O	O
in	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

Sublethal	NN	O	O
levels	NN	O	O
of	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
stimulate	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
suppress	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Sublethal	NN	O	O
levels	NN	O	O
of	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
are	NN	O	O
well	NN	O	O
known	NN	O	O
to	NN	O	O
alter	NN	O	O
T	NN	O	O
cell	NN	O	O
functional	NN	O	O
responses	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
underlying	NN	O	O
mechanisms	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

The	NN	O	O
current	NN	O	O
study	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
on	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
mediated	NN	O	O
by	NN	O	O
c-Fos/c-Jun	NN	O	B-protein
AP-1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
acutely	NN	O	O
exposed	NN	O	O
to	NN	O	O
micromolar	NN	O	O
concentrations	NN	O	O
of	NN	O	O
H2O2	NN	O	O
exhibit	NN	O	O
substantial	NN	O	O
increases	NN	O	O
in	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
but	NN	O	O
not	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

The	NN	O	O
preferential	NN	O	O
induction	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
by	NN	O	O
H2O2	NN	O	O
did	NN	O	O
not	NN	O	O
represent	NN	O	O
redox	NN	O	O
stabilization	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
transcripts	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
oxidative	NN	O	O
signals	NN	O	O
closely	NN	O	O
resembled	NN	O	O
PHA/PMA	NN	O	O
stimulation	NN	O	O
by	NN	O	O
effectively	NN	O	O
transactivating	NN	O	O
the	NN	O	O
full	NN	O	O
length	NN	O	O
c-jun	NN	O	B-DNA
promoter	NN	O	O
via	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
jun1	NN	O	I-DNA
tumor	NN	O	I-DNA
promoter-responsive	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
TRE	NN	O	I-DNA
)	NN	O	I-DNA
-like	NN	O	I-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
the	NN	O	O
complexes	NN	O	O
binding	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
AP-1	NN	O	I-DNA
TRE	NN	O	I-DNA
and	NN	O	O
jun	NN	O	B-DNA
TRE-like	NN	O	I-DNA
motifs	NN	O	I-DNA
in	NN	O	O
cells	NN	O	O
exposed	NN	O	O
to	NN	O	O
oxidative	NN	O	O
signals	NN	O	O
or	NN	O	O
PHA/PMA	NN	O	O
were	NN	O	O
indistinguishable	NN	O	O
,	NN	O	O
being	NN	O	O
composed	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
and	NN	O	O
JunD	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
PHA/PMA	NN	O	O
but	NN	O	O
not	NN	O	O
oxidative	NN	O	O
signals	NN	O	O
induced	NN	O	O
the	NN	O	O
coordinate	NN	O	O
activation	NN	O	O
of	NN	O	O
reporter	NN	O	B-DNA
constructs	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
AP-1-TRE	NN	O	B-DNA
,	NN	O	I-DNA
NF-AT	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
IL-2	NN	O	I-DNA
promoter	NN	O	I-DNA
regions	NN	O	I-DNA
along	NN	O	O
with	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
sublethal	NN	O	O
levels	NN	O	O
of	NN	O	O
H2O2	NN	O	O
actively	NN	O	O
suppressed	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-DNA
reporters	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
cells	NN	O	O
stimulated	NN	O	O
with	NN	O	O
PHA/PMA	NN	O	O
.	NN	O	O

Gel	NN	O	O
shift	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
oxidative	NN	O	O
suppression	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
represented	NN	O	O
inhibition	NN	O	O
in	NN	O	O
the	NN	O	O
early	NN	O	O
generation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
complexes	NN	O	I-protein
rather	NN	O	O
than	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
preformed	NN	O	O
NF-AT	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
oxidative	NN	O	O
signals	NN	O	O
can	NN	O	O
positively	NN	O	O
and	NN	O	O
negatively	NN	O	O
regulate	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
transcriptional	NN	O	O
events	NN	O	O
and	NN	O	O
that	NN	O	O
changes	NN	O	O
in	NN	O	O
cellular	NN	O	O
redox	NN	O	O
can	NN	O	O
uncouple	NN	O	O
AP-1	NN	O	B-protein
regulation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
from	NN	O	O
transcriptional	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
via	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Inorganic	NN	O	O
lead	NN	O	O
activates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Inorganic	NN	O	O
lead	NN	O	O
(	NN	O	O
Pb	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
ubiquitous	NN	O	O
environmental	NN	O	O
contaminant	NN	O	O
that	NN	O	O
produces	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
effects	NN	O	O
on	NN	O	O
humoral	NN	O	O
and	NN	O	O
cell	NN	O	O
mediated	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

The	NN	O	O
underlying	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
for	NN	O	O
Pb	NN	O	O
's	NN	O	O
complex	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
remain	NN	O	O
obscure	NN	O	O
.	NN	O	O

Many	NN	O	O
of	NN	O	O
Pb	NN	O	O
's	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
could	NN	O	O
be	NN	O	O
explained	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
critical	NN	O	O
for	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
function	NN	O	O
and	NN	O	O
is	NN	O	O
a	NN	O	O
strong	NN	O	O
inducer	NN	O	O
of	NN	O	O
HIV-LTR	NN	O	B-DNA
activation	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Pb	NN	O	O
at	NN	O	O
physiologically	NN	O	O
relevant	NN	O	O
concentrations	NN	O	O
activates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Pb-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
antibodies	NN	O	B-protein
for	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
p50	NN	O	B-protein
subunits	NN	O	O
but	NN	O	O
not	NN	O	O
cRel	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
:	NN	O	I-protein
p50	NN	O	I-protein
heterodimer	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
is	NN	O	O
involved	NN	O	O
.	NN	O	O

Functional	NN	O	O
activation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
Pb	NN	O	O
was	NN	O	O
confirmed	NN	O	O
using	NN	O	O
primary	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
transfected	NN	O	O
with	NN	O	O
an	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
dependent	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
construct	NN	O	I-DNA
.	NN	O	O

Pb	NN	O	O
did	NN	O	O
not	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
4	NN	O	O
different	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
lymphoid	NN	O	O
cell	NN	O	O
lines	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
reliable	NN	O	O
surrogates	NN	O	O
for	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
molecular	NN	O	O
mediator	NN	O	O
of	NN	O	O
Pb-induced	NN	O	O
immunotoxicity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
cAMP-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
activities	NN	O	O
in	NN	O	O
CD40-activated	NN	O	B-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
,	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
undergo	NN	O	O
long-term	NN	O	O
proliferation	NN	O	O
when	NN	O	O
activated	NN	O	O
through	NN	O	O
CD40	NN	O	B-protein
,	NN	O	O
a	NN	O	O
protein	NN	O	O
expressed	NN	O	O
on	NN	O	O
their	NN	O	O
cell	NN	O	O
surface	NN	O	O
.	NN	O	O

The	NN	O	O
nature	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
-dependent	NN	O	O
signals	NN	O	O
in	NN	O	O
proliferating	NN	O	O
fresh	NN	O	O
human	NN	O	B-cell_line
Epstein-Barr	NN	O	I-cell_line
virus-negative	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
is	NN	O	O
currently	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
a	NN	O	O
CD40	NN	O	B-protein
-dependent	NN	O	O
B	NN	O	O
cell	NN	O	O
culture	NN	O	O
system	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
examine	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
different	NN	O	O
signal	NN	O	B-DNA
transduction	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Protein	NN	O	O
kinase	NN	O	O
C	NN	O	O
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
depletion	NN	O	O
generated	NN	O	O
by	NN	O	O
a	NN	O	O
long-term	NN	O	O
phorbol	NN	O	O
12	NN	O	O
myristate	NN	O	O
13-acetate	NN	O	O
treatment	NN	O	O
had	NN	O	O
weak	NN	O	O
effects	NN	O	O
on	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
directly	NN	O	O
involved	NN	O	O
in	NN	O	O
mediating	NN	O	O
CD40	NN	O	B-protein
-dependent	NN	O	O
signals	NN	O	O
.	NN	O	O

The	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
PTK	NN	O	B-protein
)	NN	O	O
-specific	NN	O	O
inhibitor	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
dramatically	NN	O	O
decreased	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
without	NN	O	O
altering	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus-1	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
a	NN	O	O
promoter	NN	O	B-DNA
largely	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
cAMP-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
specific	NN	O	O
inhibitor	NN	O	O
H-89	NN	O	O
totally	NN	O	O
inhibited	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
activity	NN	O	O
at	NN	O	O
a	NN	O	O
concentration	NN	O	O
as	NN	O	O
low	NN	O	O
as	NN	O	O
100	NN	O	O
nM	NN	O	O
without	NN	O	O
affecting	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
and	NN	O	O
supershift	NN	O	O
assay	NN	O	O
using	NN	O	O
an	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sequence	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
kappa	NN	O	B-protein
light	NN	O	I-protein
chain	NN	O	I-protein
as	NN	O	O
a	NN	O	O
probe	NN	O	O
,	NN	O	O
revealed	NN	O	O
that	NN	O	O
both	NN	O	O
p65	NN	O	B-protein
(	NN	O	O
RelA	NN	O	B-protein
)	NN	O	O
and	NN	O	O
c-Rel	NN	O	B-protein
were	NN	O	O
present	NN	O	O
in	NN	O	O
CD40-stimulated	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

While	NN	O	O
PKC	NN	O	B-protein
depletion	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
level	NN	O	O
,	NN	O	O
treatment	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
H-89	NN	O	O
or	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
provoked	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
level	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
establish	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
different	NN	O	O
signal	NN	O	O
transducing	NN	O	O
pathways	NN	O	O
leading	NN	O	O
to	NN	O	O
CD40	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Interferons	NN	O	B-protein
induce	NN	O	O
normal	NN	O	O
and	NN	O	O
aberrant	NN	O	O
retinoic-acid	NN	O	B-protein
receptors	NN	O	I-protein
type	NN	O	I-protein
alpha	NN	O	I-protein
in	NN	O	O
acute	NN	O	B-cell_type
promyelocytic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
potentiation	NN	O	O
of	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
retinoid-dependent	NN	O	O
differentiation	NN	O	B-protein
markers	NN	O	I-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
the	NN	O	O
acute	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
(	NN	O	I-cell_line
APL	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
NB4	NN	O	B-cell_line
with	NN	O	O
interferon	NN	O	B-protein
alpha	NN	O	I-protein
(	NN	O	O
IFN	NN	O	B-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
IFN	NN	O	B-protein
(	NN	O	I-protein
beta	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
gamma	NN	O	I-protein
,	NN	O	O
results	NN	O	O
in	NN	O	O
an	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
transcripts	NN	O	O
coding	NN	O	O
for	NN	O	O
retinoic-acid	NN	O	B-protein
receptor	NN	O	I-protein
type	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
leukemia-specific	NN	O	B-protein
retinoic	NN	O	I-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
PML-RAR	NN	O	B-protein
.	NN	O	O

Transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
RAR	NN	O	B-RNA
(	NN	O	I-RNA
alpha	NN	O	I-RNA
)	NN	O	I-RNA
and	NN	O	I-RNA
PML-RAR	NN	O	I-RNA
mRNAs	NN	O	I-RNA
is	NN	O	O
rapid	NN	O	O
and	NN	O	O
it	NN	O	O
is	NN	O	O
parallelled	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
corresponding	NN	O	O
proteins	NN	O	O
.	NN	O	O

Up-regulation	NN	O	O
of	NN	O	O
RAR	NN	O	B-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
PML-RAR	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
IFN	NN	O	B-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
strong	NN	O	O
potentiation	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
2	NN	O	O
retinoid-dependent	NN	O	B-protein
granulocytic	NN	O	I-protein
markers	NN	O	I-protein
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
granulocyte-colony-stimulating	NN	O	B-RNA
factor	NN	O	I-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
leukocyte	NN	O	B-protein
alkaline	NN	O	I-protein
phosphatase	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
IFN	NN	O	B-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
does	NN	O	O
not	NN	O	O
have	NN	O	O
any	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
retinoid-dependent	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
myeloid	NN	O	B-protein
surface	NN	O	I-protein
markers	NN	O	I-protein
CD11b	NN	O	B-protein
and	NN	O	O
CD33	NN	O	B-protein
.	NN	O	O

The	NN	O	O
IFN	NN	O	B-protein
-dependent	NN	O	O
increase	NN	O	O
in	NN	O	O
RAR	NN	O	B-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
levels	NN	O	O
and	NN	O	O
the	NN	O	O
enhancing	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
on	NN	O	O
retinoid-dependent	NN	O	B-protein
granulocytic	NN	O	I-protein
markers	NN	O	I-protein
expression	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
characteristic	NN	O	O
of	NN	O	O
PML-RAR	NN	O	B-cell_line
positive	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
since	NN	O	O
the	NN	O	O
phenomena	NN	O	O
are	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
HL-60	NN	O	B-cell_line
promyelocytes	NN	O	I-cell_line
.	NN	O	O

Interferons	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
retinoids	NN	O	O
inhibit	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
the	NN	O	O
2	NN	O	O
classes	NN	O	O
of	NN	O	O
compounds	NN	O	O
do	NN	O	O
not	NN	O	O
significantly	NN	O	O
interact	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
anti-proliferative	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
the	NN	O	O
possible	NN	O	O
use	NN	O	O
of	NN	O	O
combinations	NN	O	O
between	NN	O	O
IFNs	NN	O	B-protein
and	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
in	NN	O	O
the	NN	O	O
cyto-differentiating	NN	O	O
treatment	NN	O	O
of	NN	O	O
APL	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Modulatory	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
catecholamines	NN	O	O
on	NN	O	O
human	NN	O	O
interleukin	NN	O	B-protein
-12	NN	O	O
and	NN	O	O
interleukin	NN	O	B-protein
-10	NN	O	O
production	NN	O	O
:	NN	O	O
clinical	NN	O	O
implications	NN	O	O
.	NN	O	O

Interleukin-12	NN	O	B-protein
(	NN	O	O
IL-12	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
key	NN	O	O
inducer	NN	O	O
of	NN	O	O
differentiation	NN	O	O
of	NN	O	O
uncommitted	NN	O	B-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
(	NN	O	I-cell_type
TH	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
toward	NN	O	O
the	NN	O	O
TH1	NN	O	O
phenotype	NN	O	O
,	NN	O	O
which	NN	O	O
regulates	NN	O	O
cellular	NN	O	O
immunity	NN	O	O
,	NN	O	O
whereas	NN	O	O
IL-10	NN	O	B-protein
inhibits	NN	O	O
TH1	NN	O	O
functions	NN	O	O
and	NN	O	O
potentiates	NN	O	O
TH2-regulated	NN	O	O
responses	NN	O	O
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
humoral	NN	O	O
immunity	NN	O	O
)	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
potential	NN	O	O
effects	NN	O	O
of	NN	O	O
stress	NN	O	O
on	NN	O	O
TH1/TH2	NN	O	O
balance	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
three	NN	O	O
prototype	NN	O	O
stress	NN	O	O
hormones-dexamethasone	NN	O	O
(	NN	O	O
a	NN	O	O
synthetic	NN	O	O
glucocorticoid	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
catecholamines	NN	O	O
norepinephrine	NN	O	O
and	NN	O	O
epinephrine-to	NN	O	O
alter	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
(	NN	O	O
p70	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IL-10	NN	O	B-protein
induced	NN	O	O
by	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
in	NN	O	O
human	NN	O	O
whole	NN	O	O
blood	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
inhibited	NN	O	O
LPS-induced	NN	O	O
bioactive	NN	O	O
IL-12	NN	O	B-protein
production	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
fashion	NN	O	O
and	NN	O	O
at	NN	O	O
physiologically	NN	O	O
relevant	NN	O	O
concentrations	NN	O	O
;	NN	O	O
it	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
IL-10	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

The	NN	O	O
glucocorticoid-induced	NN	O	O
reduction	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
production	NN	O	O
was	NN	O	O
antagonized	NN	O	O
by	NN	O	O
RU	NN	O	O
486	NN	O	O
,	NN	O	O
a	NN	O	O
glucocorticoid-receptor	NN	O	O
antagonist	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
it	NN	O	O
was	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
.	NN	O	O

Norepinephrine	NN	O	O
and	NN	O	O
epinephrine	NN	O	O
also	NN	O	O
suppressed	NN	O	O
IL-12	NN	O	B-protein
production	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
fashion	NN	O	O
and	NN	O	O
at	NN	O	O
physiological	NN	O	O
concentrations	NN	O	O
;	NN	O	O
both	NN	O	O
catecholamines	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
dose-dependently	NN	O	O
increased	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
either	NN	O	O
catecholamine	NN	O	O
on	NN	O	O
IL-12	NN	O	B-protein
or	NN	O	O
IL-10	NN	O	B-protein
secretion	NN	O	O
were	NN	O	O
blocked	NN	O	O
completely	NN	O	O
by	NN	O	O
propranolol	NN	O	O
,	NN	O	O
a	NN	O	O
beta-adrenoreceptor	NN	O	O
antagonist	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
they	NN	O	O
were	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
beta-adrenergic	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
may	NN	O	O
regulate	NN	O	O
IL-12	NN	O	O
and	NN	O	O
IL-10	NN	O	O
secretion	NN	O	O
and	NN	O	O
,	NN	O	O
hence	NN	O	O
,	NN	O	O
TH1/TH2	NN	O	O
balance	NN	O	O
via	NN	O	O
the	NN	O	O
peripheral	NN	O	O
end-effectors	NN	O	O
of	NN	O	O
the	NN	O	O
stress	NN	O	O
system	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
stress	NN	O	O
may	NN	O	O
cause	NN	O	O
a	NN	O	O
selective	NN	O	O
suppression	NN	O	O
of	NN	O	O
TH1	NN	O	O
functions	NN	O	O
and	NN	O	O
a	NN	O	O
shift	NN	O	O
toward	NN	O	O
a	NN	O	O
TH2	NN	O	B-protein
cytokine	NN	O	I-protein
pattern	NN	O	O
rather	NN	O	O
than	NN	O	O
generalized	NN	O	O
TH	NN	O	O
suppression	NN	O	O
.	NN	O	O

The	NN	O	O
TH1-to-TH2	NN	O	O
shift	NN	O	O
may	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
stress-induced	NN	O	O
susceptibility	NN	O	O
of	NN	O	O
the	NN	O	O
organism	NN	O	O
to	NN	O	O
certain	NN	O	O
infections	NN	O	O
.	NN	O	O

Through	NN	O	O
the	NN	O	O
same	NN	O	O
or	NN	O	O
a	NN	O	O
reciprocal	NN	O	O
mechanism	NN	O	O
,	NN	O	O
states	NN	O	O
associated	NN	O	O
with	NN	O	O
chronic	NN	O	O
hyperactivity	NN	O	O
or	NN	O	O
hypoactivity	NN	O	O
of	NN	O	O
the	NN	O	O
stress	NN	O	O
system	NN	O	O
might	NN	O	O
influence	NN	O	O
the	NN	O	O
susceptibility	NN	O	O
of	NN	O	O
an	NN	O	O
individual	NN	O	O
to	NN	O	O
certain	NN	O	O
autoimmune	NN	O	O
,	NN	O	O
allergic	NN	O	O
,	NN	O	O
infectious	NN	O	O
,	NN	O	O
or	NN	O	O
neoplastic	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
Treponema	NN	O	B-protein
pallidum	NN	O	I-protein
and	NN	O	I-protein
Borrelia	NN	O	I-protein
burgdorferi	NN	O	I-protein
lipoproteins	NN	O	I-protein
and	NN	O	O
synthetic	NN	O	O
lipopeptides	NN	O	O
proceeds	NN	O	O
via	NN	O	O
a	NN	O	O
pathway	NN	O	O
distinct	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
lipopolysaccharide	NN	O	O
but	NN	O	O
involves	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

There	NN	O	O
is	NN	O	O
increasing	NN	O	O
evidence	NN	O	O
that	NN	O	O
lipoproteins	NN	O	B-protein
of	NN	O	O
Treponema	NN	O	O
pallidum	NN	O	O
and	NN	O	O
Borrelia	NN	O	O
burgdorferi	NN	O	O
are	NN	O	O
key	NN	O	O
inflammatory	NN	O	O
mediators	NN	O	O
during	NN	O	O
syphilis	NN	O	O
and	NN	O	O
Lyme	NN	O	O
disease	NN	O	O
.	NN	O	O

A	NN	O	O
principal	NN	O	O
objective	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
identify	NN	O	O
more	NN	O	O
precisely	NN	O	O
similarities	NN	O	O
and	NN	O	O
divergences	NN	O	O
among	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-	NN	O	O
and	NN	O	O
lipoprotein-lipopeptide-induced	NN	O	O
immune	NN	O	O
cell	NN	O	O
signaling	NN	O	O
events	NN	O	O
.	NN	O	O

Like	NN	O	O
LPS	NN	O	O
,	NN	O	O
purified	NN	O	B-protein
native	NN	O	I-protein
B.	NN	O	I-protein
burgdorferi	NN	O	I-protein
OspA	NN	O	I-protein
and	NN	O	O
synthetic	NN	O	O
analogs	NN	O	O
of	NN	O	O
OspA	NN	O	B-protein
,	NN	O	O
OspB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
two	NN	O	O
T.	NN	O	B-protein
pallidum	NN	O	I-protein
lipoproteins	NN	O	I-protein
(	NN	O	O
Tpp47	NN	O	B-protein
and	NN	O	O
Tpp17	NN	O	B-protein
)	NN	O	O
all	NN	O	O
induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Acylation	NN	O	O
of	NN	O	O
OspA	NN	O	B-protein
and	NN	O	O
the	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
was	NN	O	O
requisite	NN	O	O
for	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Polymyxin	NN	O	O
B	NN	O	O
abrogated	NN	O	O
only	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
70Z/3-derived	NN	O	B-cell_line
pre-B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
either	NN	O	O
lacking	NN	O	O
or	NN	O	O
expressing	NN	O	O
human	NN	O	B-protein
CD14	NN	O	I-protein
(	NN	O	O
the	NN	O	O
LPS	NN	O	B-protein
receptor	NN	O	I-protein
)	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
observed	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-protein
CD14	NN	O	I-protein
imparted	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
LPS	NN	O	O
but	NN	O	O
not	NN	O	O
to	NN	O	O
OspA	NN	O	B-protein
or	NN	O	O
spirochetal	NN	O	B-protein
lipopeptides	NN	O	I-protein
(	NN	O	O
assessed	NN	O	O
by	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
expression	NN	O	O
of	NN	O	O
surface	NN	O	B-protein
immunoglobulin	NN	O	I-protein
M	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
biological	NN	O	O
relevance	NN	O	O
of	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
T.	NN	O	B-protein
pallidum	NN	O	I-protein
lipoproteins	NN	O	I-protein
-lipopeptides	NN	O	O
induce	NN	O	O
both	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
was	NN	O	O
supported	NN	O	O
by	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
synthetic	NN	O	O
analogs	NN	O	O
to	NN	O	O
promote	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
replication	NN	O	O
in	NN	O	O
chronically	NN	O	O
infected	NN	O	O
U1	NN	O	O
monocytoid	NN	O	O
cells	NN	O	O
;	NN	O	O
these	NN	O	O
observations	NN	O	O
also	NN	O	O
suggest	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
whereby	NN	O	O
a	NN	O	O
syphilitic	NN	O	O
chancre	NN	O	O
can	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
cofactor	NN	O	O
for	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
transmission	NN	O	O
.	NN	O	O

The	NN	O	O
combined	NN	O	O
data	NN	O	O
lend	NN	O	O
additional	NN	O	O
support	NN	O	O
to	NN	O	O
the	NN	O	O
proposal	NN	O	O
that	NN	O	O
spirochetal	NN	O	B-protein
lipoproteins	NN	O	I-protein
and	NN	O	O
LPS	NN	O	O
initiate	NN	O	O
monocyte	NN	O	O
activation	NN	O	O
via	NN	O	O
different	NN	O	O
cell	NN	O	O
surface	NN	O	O
events	NN	O	O
but	NN	O	O
that	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
ultimately	NN	O	O
converge	NN	O	O
to	NN	O	O
produce	NN	O	O
qualitatively	NN	O	O
similar	NN	O	O
cellular	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

LYSP100	NN	O	B-protein
-associated	NN	O	O
nuclear	NN	O	O
domains	NN	O	O
(	NN	O	O
LANDs	NN	O	O
)	NN	O	O
:	NN	O	O
description	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
class	NN	O	O
of	NN	O	O
subnuclear	NN	O	O
structures	NN	O	O
and	NN	O	O
their	NN	O	O
relationship	NN	O	O
to	NN	O	O
PML	NN	O	O
nuclear	NN	O	O
bodies	NN	O	O
.	NN	O	O

The	NN	O	O
PML	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
fused	NN	O	O
to	NN	O	O
the	NN	O	O
retinoic	NN	O	B-DNA
acid	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
RAR	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
t	NN	O	O
(	NN	O	O
15	NN	O	O
;	NN	O	O
17	NN	O	O
)	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
,	NN	O	O
creating	NN	O	O
a	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
fusion	NN	O	I-protein
oncoprotein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
PML	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
has	NN	O	O
been	NN	O	O
localized	NN	O	O
to	NN	O	O
subnuclear	NN	O	B-cell_line
dot-like	NN	O	I-cell_line
structures	NN	O	I-cell_line
variously	NN	O	O
termed	NN	O	O
PODs	NN	O	O
,	NN	O	O
ND10s	NN	O	O
,	NN	O	O
Kr	NN	O	O
bodies	NN	O	O
,	NN	O	O
or	NN	O	O
PML	NN	O	O
nuclear	NN	O	O
bodies	NN	O	O
(	NN	O	O
PML	NN	O	O
NBs	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
describes	NN	O	O
the	NN	O	O
cloning	NN	O	O
of	NN	O	O
a	NN	O	O
lymphoid-restricted	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
LYSP100	NN	O	B-DNA
,	NN	O	O
that	NN	O	O
is	NN	O	O
homologous	NN	O	O
to	NN	O	O
another	NN	O	O
protein	NN	O	O
that	NN	O	O
localizes	NN	O	O
to	NN	O	O
PML	NN	O	O
NBs	NN	O	O
,	NN	O	O
SP100	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
SP100	NN	O	B-DNA
homology	NN	O	I-DNA
regions	NN	O	I-DNA
,	NN	O	O
one	NN	O	O
LYSP100	NN	O	B-DNA
cDNA	NN	O	I-DNA
isoform	NN	O	I-DNA
contains	NN	O	O
a	NN	O	O
bromodomain	NN	O	B-protein
and	NN	O	O
a	NN	O	O
PHD/TTC	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
which	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

By	NN	O	O
immunofluorescence	NN	O	O
,	NN	O	O
LYSP100	NN	O	B-DNA
was	NN	O	O
localized	NN	O	O
to	NN	O	O
nuclear	NN	O	O
dots	NN	O	O
that	NN	O	O
were	NN	O	O
surprisingly	NN	O	O
largely	NN	O	O
nonoverlapping	NN	O	O
with	NN	O	O
PML	NN	O	O
NBs	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
minority	NN	O	O
of	NN	O	O
LYSP100	NN	O	B-DNA
nuclear	NN	O	O
dots	NN	O	O
exactly	NN	O	O
colocalized	NN	O	O
with	NN	O	O
PML	NN	O	B-cell_type
and	NN	O	O
SP100	NN	O	B-protein
.	NN	O	O

We	NN	O	O
term	NN	O	O
the	NN	O	O
LYSP100	NN	O	B-DNA
structures	NN	O	O
``	NN	O	O
LANDs	NN	O	O
,	NN	O	O
''	NN	O	O
for	NN	O	O
LYSP100	NN	O	B-DNA
-associated	NN	O	O
nuclear	NN	O	O
domains	NN	O	O
.	NN	O	O

Although	NN	O	O
LYSP100	NN	O	B-DNA
is	NN	O	O
expressed	NN	O	O
only	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
LANDs	NN	O	O
could	NN	O	O
be	NN	O	O
visualized	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
transfection	NN	O	O
of	NN	O	O
a	NN	O	O
LYSP100	NN	O	B-DNA
cDNA	NN	O	I-DNA
.	NN	O	O

Immunoelectron	NN	O	O
microscopy	NN	O	O
revealed	NN	O	O
LANDs	NN	O	O
to	NN	O	O
be	NN	O	O
globular	NN	O	O
,	NN	O	O
electron-dense	NN	O	O
structures	NN	O	O
morphologically	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
annular	NN	O	O
structures	NN	O	O
characteristic	NN	O	O
of	NN	O	O
PML	NN	O	O
NBs	NN	O	O
.	NN	O	O

LANDs	NN	O	O
were	NN	O	O
most	NN	O	O
often	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
nucleoplasm	NN	O	O
,	NN	O	O
but	NN	O	O
were	NN	O	O
also	NN	O	O
found	NN	O	O
at	NN	O	O
the	NN	O	O
nuclear	NN	O	O
membrane	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
structures	NN	O	O
may	NN	O	O
traffic	NN	O	O
between	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
and	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

By	NN	O	O
double-immunogold	NN	O	O
labeling	NN	O	O
of	NN	O	O
PML	NN	O	B-cell_type
and	NN	O	O
LYSP100	NN	O	B-DNA
,	NN	O	O
some	NN	O	O
LANDs	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
contain	NN	O	O
both	NN	O	O
PML	NN	O	B-cell_type
and	NN	O	O
LYSP100	NN	O	B-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
PML	NN	O	B-cell_type
is	NN	O	O
localized	NN	O	O
to	NN	O	O
a	NN	O	O
second	NN	O	O
subnuclear	NN	O	O
domain	NN	O	O
that	NN	O	O
is	NN	O	O
morphologically	NN	O	O
and	NN	O	O
biochemically	NN	O	O
distinct	NN	O	O
from	NN	O	O
PML	NN	O	B-cell_type
NBs	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
steroid-regulated	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
thymoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Early	NN	O	O
studies	NN	O	O
in	NN	O	O
murine	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
indicated	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
transactivation	NN	O	O
functions	NN	O	O
encoded	NN	O	O
in	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
GR	NN	O	I-protein
)	NN	O	I-protein
N-terminal	NN	O	I-protein
domain	NN	O	I-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
glucocorticoid-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
more	NN	O	O
recent	NN	O	O
studies	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
domain	NN	O	I-protein
is	NN	O	O
not	NN	O	O
necessary	NN	O	O
for	NN	O	O
steroid-regulated	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
that	NN	O	O
GR	NN	O	B-protein
-mediated	NN	O	O
transrepression	NN	O	O
may	NN	O	O
be	NN	O	O
the	NN	O	O
more	NN	O	O
critical	NN	O	O
mechanism	NN	O	O
.	NN	O	O

To	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
N-terminal	NN	O	I-protein
transactivation	NN	O	I-protein
domain	NN	O	I-protein
in	NN	O	O
mediating	NN	O	O
murine	NN	O	O
thymocyte	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
we	NN	O	O
stably	NN	O	O
transfected	NN	O	O
GR	NN	O	B-protein
,	NN	O	O
GR	NN	O	B-protein
variants	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
AR	NN	O	B-protein
)	NN	O	O
into	NN	O	O
receptor-negative	NN	O	B-cell_line
S49	NN	O	I-cell_line
murine	NN	O	I-cell_line
thymoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

GR	NN	O	B-protein
expression	NN	O	O
levels	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
rate-limiting	NN	O	O
for	NN	O	O
initiating	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
pathway	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
positive	NN	O	O
correlation	NN	O	O
between	NN	O	O
steroid	NN	O	O
sensitivity	NN	O	O
and	NN	O	O
GR	NN	O	B-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
an	NN	O	O
integrated	NN	O	O
mouse	NN	O	B-DNA
mammary	NN	O	I-DNA
tumor	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
MMTV	NN	O	I-DNA
)	NN	O	I-DNA
LTR	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
chimeric	NN	O	I-protein
receptors	NN	O	I-protein
containing	NN	O	O
the	NN	O	O
potent	NN	O	O
VP16	NN	O	B-DNA
and	NN	O	I-DNA
E1A	NN	O	I-DNA
viral	NN	O	I-DNA
transactivation	NN	O	I-DNA
domains	NN	O	I-DNA
in	NN	O	O
place	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
N	NN	O	I-protein
terminus	NN	O	I-protein
revealed	NN	O	O
that	NN	O	O
even	NN	O	O
low	NN	O	O
level	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
receptors	NN	O	O
resulted	NN	O	O
in	NN	O	O
both	NN	O	O
enhanced	NN	O	O
steroid	NN	O	O
sensitivity	NN	O	O
and	NN	O	O
MMTV	NN	O	O
induction	NN	O	O
,	NN	O	O
thus	NN	O	O
supporting	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
transactivation	NN	O	O
in	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
AR	NN	O	B-protein
can	NN	O	O
initiate	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
S49	NN	O	B-cell_line
cells	NN	O	I-cell_line
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
5	NN	O	O
alpha-dihydrotestosterone	NN	O	O
,	NN	O	O
despite	NN	O	O
its	NN	O	O
relative	NN	O	O
inability	NN	O	O
to	NN	O	O
induce	NN	O	O
high	NN	O	O
level	NN	O	O
expression	NN	O	O
of	NN	O	O
MMTV	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
this	NN	O	O
further	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
steroid-regulated	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
endogenous	NN	O	O
thymocyte-specific	NN	O	B-DNA
gene	NN	O	I-DNA
called	NN	O	O
GIG18	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
GIG18	NN	O	B-DNA
was	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
to	NN	O	O
comparable	NN	O	O
levels	NN	O	O
by	NN	O	O
both	NN	O	O
AR	NN	O	B-protein
and	NN	O	O
GR	NN	O	B-protein
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
AR	NN	O	B-protein
can	NN	O	O
indeed	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
in	NN	O	O
S49	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
,	NN	O	O
moreover	NN	O	O
,	NN	O	O
that	NN	O	O
GIG18	NN	O	B-DNA
induction	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
marker	NN	O	O
of	NN	O	O
early	NN	O	O
apoptotic	NN	O	O
events	NN	O	O
in	NN	O	O
steroid-treated	NN	O	O
cells	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
support	NN	O	O
our	NN	O	O
conclusion	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
transactivation	NN	O	O
is	NN	O	O
a	NN	O	O
necessary	NN	O	O
signaling	NN	O	O
component	NN	O	O
of	NN	O	O
S49	NN	O	O
cell	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
although	NN	O	O
an	NN	O	O
additional	NN	O	O
role	NN	O	O
for	NN	O	O
GR	NN	O	B-protein
-mediated	NN	O	O
transrepression	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
excluded	NN	O	O
.	NN	O	O

-DOCSTART-	O

Multiple	NN	O	O
p21ras	NN	O	B-protein
effector	NN	O	O
pathways	NN	O	O
regulate	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
Nuclear	NN	O	B-protein
Factor	NN	O	I-protein
of	NN	O	I-protein
Activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
target	NN	O	O
for	NN	O	O
p21ras	NN	O	B-protein
and	NN	O	O
calcium	NN	O	O
signalling	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
p21ras	NN	O	B-protein
signals	NN	O	O
acting	NN	O	O
in	NN	O	O
synergy	NN	O	O
with	NN	O	O
calcium	NN	O	O
/calcineurin	NN	O	B-protein
signals	NN	O	O
.	NN	O	O

One	NN	O	O
p21ras	NN	O	B-protein
effector	NN	O	O
pathway	NN	O	O
involves	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
ERK-2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
NFAT	NN	O	B-protein
regulation	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
MAPKK-1	NN	O	I-protein
prevents	NN	O	O
NFAT	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

Constitutively	NN	O	O
active	NN	O	O
MAPKK-1	NN	O	B-protein
fully	NN	O	O
activates	NN	O	O
ERK-2	NN	O	B-protein
and	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Elk-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
does	NN	O	O
not	NN	O	O
substitute	NN	O	O
for	NN	O	O
activated	NN	O	O
p21ras	NN	O	B-protein
and	NN	O	O
synergize	NN	O	O
with	NN	O	O
calcium	NN	O	O
/calcineurin	NN	O	B-protein
signals	NN	O	O
to	NN	O	O
induce	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
N17Rac	NN	O	I-protein
also	NN	O	O
prevents	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
p21ras	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
but	NN	O	O
without	NN	O	O
interfering	NN	O	O
with	NN	O	O
the	NN	O	O
ERK-2	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
complex	NN	O	O
comprising	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
group	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
by	NN	O	O
p21ras	NN	O	B-protein
also	NN	O	O
requires	NN	O	O
Rac-1	NN	O	B-protein
function	NN	O	O
.	NN	O	O

Activated	NN	O	B-protein
Rac-1	NN	O	I-protein
could	NN	O	O
mimic	NN	O	O
activated	NN	O	B-protein
p21ras	NN	O	I-protein
to	NN	O	O
induce	NN	O	O
AP-1	NN	O	B-protein
but	NN	O	O
not	NN	O	O
to	NN	O	O
induce	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
activated	NN	O	O
MAPKK-1	NN	O	B-protein
and	NN	O	O
Rac-1	NN	O	B-protein
could	NN	O	O
not	NN	O	O
substitute	NN	O	O
for	NN	O	O
activated	NN	O	O
p21ras	NN	O	B-protein
and	NN	O	O
synergize	NN	O	O
with	NN	O	O
calcium	NN	O	O
signals	NN	O	O
to	NN	O	O
induce	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
p21ras	NN	O	B-protein
regulation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
multiple	NN	O	O
effector	NN	O	O
pathways	NN	O	O
including	NN	O	O
those	NN	O	O
regulated	NN	O	O
by	NN	O	O
MAPKK-1	NN	O	B-protein
/ERK-2	NN	O	B-protein
and	NN	O	O
Rac-1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Retinoid	NN	O	O
differentiation	NN	O	O
therapy	NN	O	O
in	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
specific	NN	O	O
type	NN	O	O
of	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
morphology	NN	O	O
of	NN	O	O
the	NN	O	O
blast	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
specific	NN	O	O
t	NN	O	O
(	NN	O	O
15	NN	O	O
;	NN	O	O
17	NN	O	O
)	NN	O	O
translocation	NN	O	O
,	NN	O	O
and	NN	O	O
risks	NN	O	O
of	NN	O	O
definite	NN	O	O
coagulopathy	NN	O	O
.	NN	O	O

Recently	NN	O	O
this	NN	O	O
leukemia	NN	O	O
was	NN	O	O
further	NN	O	O
characterized	NN	O	O
by	NN	O	O
an	NN	O	O
exquisite	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
's	NN	O	O
differentiation	NN	O	O
effect	NN	O	O
and	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
a	NN	O	O
fusion	NN	O	B-DNA
gene	NN	O	I-DNA
altering	NN	O	O
the	NN	O	O
gene	NN	O	O
of	NN	O	O
RARalpha	NN	O	B-protein
and	NN	O	O
a	NN	O	O
novel	NN	O	B-DNA
gene	NN	O	I-DNA
PML	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
differentiation	NN	O	O
therapy	NN	O	O
with	NN	O	O
retinoids	NN	O	O
in	NN	O	O
APL	NN	O	O
patients	NN	O	O
follows	NN	O	O
strict	NN	O	O
guidelines	NN	O	O
related	NN	O	O
both	NN	O	O
to	NN	O	O
the	NN	O	O
APL	NN	O	O
cell	NN	O	O
and	NN	O	O
the	NN	O	O
biodisposal	NN	O	O
of	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
utilization	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
-signal	NN	O	O
transducer	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IFN-alpha	NN	O	B-protein
.	NN	O	O

IL-2-	NN	O	O
,	NN	O	O
IL-12-	NN	O	O
,	NN	O	O
and	NN	O	O
IFN-alpha-mediated	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
were	NN	O	O
analyzed	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
the	NN	O	O
NK3.3	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
and	NN	O	O
immunoprecipitation	NN	O	O
analyses	NN	O	O
revealed	NN	O	O
that	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
activating	NN	O	O
STAT3	NN	O	B-protein
(	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription-3	NN	O	I-protein
)	NN	O	O
and	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
IL-2	NN	O	B-protein
induced	NN	O	O
tyrosine	NN	O	O
and	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
which	NN	O	O
formed	NN	O	O
IFN-gamma-activated	NN	O	B-protein
sequence-binding	NN	O	I-protein
complexes	NN	O	I-protein
by	NN	O	O
itself	NN	O	O
and	NN	O	O
with	NN	O	O
STAT3	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IFN-alpha	NN	O	B-protein
activated	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
IL-2	NN	O	B-protein
predominantly	NN	O	O
activated	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
while	NN	O	O
IFN-alpha	NN	O	B-protein
predominantly	NN	O	O
activated	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

IL-2	NN	O	B-protein
induced	NN	O	O
less	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
IFN-alpha	NN	O	B-protein
induced	NN	O	O
greater	NN	O	O
STAT5	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
NK3.3	NN	O	B-cell_line
cells	NN	O	I-cell_line
compared	NN	O	O
with	NN	O	O
preactivated	NN	O	B-cell_line
primary	NN	O	I-cell_line
NK	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
NK3.3	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
IL-2	NN	O	B-protein
induced	NN	O	O
comparable	NN	O	O
formation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-protein
promoter	NN	O	I-protein
sis-inducible	NN	O	I-protein
element	NN	O	I-protein
IFN-gamma-activated	NN	O	I-protein
sequence-binding	NN	O	I-protein
complexes	NN	O	I-protein
containing	NN	O	O
STAT3	NN	O	B-protein
alone	NN	O	O
with	NN	O	O
complexes	NN	O	O
containing	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
while	NN	O	O
in	NN	O	O
preactivated	NN	O	B-cell_line
primary	NN	O	I-cell_line
NK	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
it	NN	O	O
preferentially	NN	O	O
induced	NN	O	O
complexes	NN	O	O
containing	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
signaling	NN	O	O
in	NN	O	O
NK3.3	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
not	NN	O	O
always	NN	O	O
identical	NN	O	O
with	NN	O	O
that	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IFN-alpha	NN	O	B-protein
,	NN	O	O
IL-12	NN	O	B-protein
induced	NN	O	O
strong	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT4	NN	O	B-protein
and	NN	O	O
variable	NN	O	O
weak	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
supershift	NN	O	O
analyses	NN	O	O
using	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
promoter	NN	O	I-DNA
sis-inducible	NN	O	I-DNA
element	NN	O	I-DNA
probe	NN	O	I-DNA
showed	NN	O	O
that	NN	O	O
IL-12	NN	O	B-protein
activated	NN	O	O
STAT4	NN	O	B-protein
,	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
and	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
induced	NN	O	O
complexes	NN	O	O
containing	NN	O	O
STAT4	NN	O	B-protein
only	NN	O	O
,	NN	O	O
STAT4	NN	O	B-protein
with	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
STAT3	NN	O	B-protein
with	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
or	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
only	NN	O	O
in	NN	O	O
preactivated	NN	O	B-cell_line
primary	NN	O	I-cell_line
NK	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

STAT1	NN	O	B-protein
alpha	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
IL-12	NN	O	B-protein
correlated	NN	O	O
with	NN	O	O
increased	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
serine	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
tyrosine	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
JAK1	NN	O	B-protein
and	NN	O	O
JAK3	NN	O	B-protein
,	NN	O	O
while	NN	O	O
IL-12	NN	O	B-protein
induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
JAK2	NN	O	B-protein
and	NN	O	O
TYK2	NN	O	B-protein
in	NN	O	O
both	NN	O	O
preactivated	NN	O	B-cell_line
primary	NN	O	I-cell_line
NK	NN	O	I-cell_line
and	NN	O	I-cell_line
NK3.3	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Differential	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
consequent	NN	O	O
differential	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
separate	NN	O	O
and	NN	O	O
overlapping	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
by	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IFN-alpha	NN	O	B-protein
may	NN	O	O
provide	NN	O	O
a	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
similarities	NN	O	O
and	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
on	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
suppression	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
function	NN	O	O
and	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
serine	NN	O	O
protease	NN	O	O
inhibitors	NN	O	O
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
N-acetylcysteine	NN	O	O
.	NN	O	O

Direct	NN	O	O
evidence	NN	O	O
that	NN	O	O
N-acetylcysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
enhances	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
presented	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
NAC	NN	O	O
blocks	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
mitogenesis	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
by	NN	O	O
protease	NN	O	O
inhibitors	NN	O	O
such	NN	O	O
as	NN	O	O
N-tosylphenylalanine	NN	O	O
chloromethyl	NN	O	O
ketone	NN	O	O
(	NN	O	O
TPCK	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
proliferative	NN	O	O
responses	NN	O	O
of	NN	O	O
purified	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
or	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
suppressed	NN	O	O
more	NN	O	O
strongly	NN	O	O
by	NN	O	O
TPCK	NN	O	O
when	NN	O	O
anti-CD28	NN	O	B-protein
rather	NN	O	O
than	NN	O	O
the	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
PMA	NN	O	O
is	NN	O	O
used	NN	O	O
as	NN	O	O
the	NN	O	O
mitogenic	NN	O	O
coactivator	NN	O	B-protein
.	NN	O	O

Cytokine	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
INF-gamma	NN	O	B-protein
)	NN	O	O
production	NN	O	O
is	NN	O	O
inhibited	NN	O	O
95-100	NN	O	O
%	NN	O	O
by	NN	O	O
concentrations	NN	O	O
of	NN	O	O
TPCK	NN	O	O
that	NN	O	O
totally	NN	O	O
suppress	NN	O	O
the	NN	O	O
mitogenesis	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
or	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
find	NN	O	O
that	NN	O	O
TPCK	NN	O	O
virtually	NN	O	O
abolishes	NN	O	O
(	NN	O	O
to	NN	O	O
less	NN	O	O
than	NN	O	O
1	NN	O	O
%	NN	O	O
)	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
but	NN	O	O
not	NN	O	O
Oct-1	NN	O	B-protein
)	NN	O	O
found	NN	O	O
in	NN	O	O
nuclear	NN	O	O
and	NN	O	O
whole	NN	O	O
cell	NN	O	O
extracts	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Strikingly	NN	O	O
,	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
effects	NN	O	O
of	NN	O	O
TPCK	NN	O	O
are	NN	O	O
blocked	NN	O	O
when	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
pretreated	NN	O	O
for	NN	O	O
15	NN	O	O
min	NN	O	O
with	NN	O	O
5	NN	O	O
mM	NN	O	O
NAC	NN	O	O
.	NN	O	O

NAC	NN	O	O
not	NN	O	O
only	NN	O	O
blocks	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
TPCK	NN	O	O
but	NN	O	O
enhances	NN	O	O
mitogenesis	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
(	NN	O	O
>	NN	O	O
2.5-fold	NN	O	O
in	NN	O	O
some	NN	O	O
cases	NN	O	O
)	NN	O	O
upon	NN	O	O
activation	NN	O	O
of	NN	O	O
unsuppressed	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
proteases	NN	O	I-protein
play	NN	O	O
obligate	NN	O	O
roles	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
mitogenesis	NN	O	O
,	NN	O	O
roles	NN	O	O
that	NN	O	O
are	NN	O	O
enhanced	NN	O	O
significantly	NN	O	O
by	NN	O	O
NAC	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signaling	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
related	NN	O	O
cytokines	NN	O	B-protein
:	NN	O	O
JAKs	NN	O	B-protein
,	NN	O	O
STATs	NN	O	B-protein
,	NN	O	O
and	NN	O	O
relationship	NN	O	O
to	NN	O	O
immunodeficiency	NN	O	O
.	NN	O	O

Cytokines	NN	O	B-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	I-protein
IL-2	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
common	NN	O	I-protein
gamma	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
gamma	NN	O	B-protein
c	NN	O	I-protein
)	NN	O	O
,	NN	O	O
including	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-7	NN	O	B-protein
,	NN	O	O
IL-9	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-15	NN	O	B-protein
,	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
monoctyes	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
cytokines	NN	O	B-protein
have	NN	O	O
overlapping	NN	O	O
biological	NN	O	O
effects	NN	O	O
that	NN	O	O
in	NN	O	O
part	NN	O	O
result	NN	O	O
from	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
shared	NN	O	B-protein
receptor	NN	O	I-protein
subunit	NN	O	I-protein
gamma	NN	O	I-protein
c	NN	O	I-protein
.	NN	O	O

Recently	NN	O	O
it	NN	O	O
has	NN	O	O
become	NN	O	O
clear	NN	O	O
that	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
activate	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
important	NN	O	O
intracellular	NN	O	B-protein
signaling	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
including	NN	O	O
the	NN	O	O
Janus	NN	O	B-protein
kinases	NN	O	I-protein
JAK1	NN	O	B-protein
and	NN	O	O
JAK3	NN	O	B-protein
and	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	O
activators	NN	O	B-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	O
STATs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
discovery	NN	O	O
of	NN	O	O
these	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
has	NN	O	O
led	NN	O	O
to	NN	O	O
important	NN	O	O
new	NN	O	O
insights	NN	O	O
into	NN	O	O
their	NN	O	O
role	NN	O	O
in	NN	O	O
lymphocyte	NN	O	O
maturation	NN	O	O
,	NN	O	O
as	NN	O	O
it	NN	O	O
has	NN	O	O
emerged	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
genes	NN	O	O
encoding	NN	O	O
both	NN	O	O
gamma	NN	O	B-protein
c	NN	O	I-protein
and	NN	O	O
JAK3	NN	O	B-protein
result	NN	O	O
in	NN	O	O
similar	NN	O	O
forms	NN	O	O
of	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
(	NN	O	O
SCID	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
we	NN	O	O
examine	NN	O	O
the	NN	O	O
structure	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
involved	NN	O	O
in	NN	O	O
their	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
discuss	NN	O	O
recent	NN	O	O
advances	NN	O	O
that	NN	O	O
have	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
how	NN	O	O
cytokines	NN	O	B-protein
elicit	NN	O	O
intracellular	NN	O	O
responses	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
their	NN	O	O
role	NN	O	O
in	NN	O	O
normal	NN	O	O
lymphoid	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cytomegalovirus	NN	O	O
modulates	NN	O	O
interleukin-6	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Complications	NN	O	O
after	NN	O	O
lung	NN	O	O
transplantation	NN	O	O
include	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
rejection	NN	O	O
and	NN	O	O
an	NN	O	O
increased	NN	O	O
incidence	NN	O	O
of	NN	O	O
infection	NN	O	O
,	NN	O	O
particularly	NN	O	O
with	NN	O	O
cytomegalovirus	NN	O	O
(	NN	O	O
CMV	NN	O	O
)	NN	O	O
.	NN	O	O

Several	NN	O	O
recent	NN	O	O
studies	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
interleukin	NN	O	O
(	NN	O	O
IL	NN	O	O
)	NN	O	O
-6	NN	O	O
may	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
detect	NN	O	O
both	NN	O	O
infection	NN	O	O
and	NN	O	O
rejection	NN	O	O
after	NN	O	O
lung	NN	O	O
transplantation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
IL-6	NN	O	B-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
bronchiolitis	NN	O	O
obliterans	NN	O	O
after	NN	O	O
transplantation	NN	O	O
.	NN	O	O

Because	NN	O	O
CMV	NN	O	O
is	NN	O	O
also	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
bronchiolitis	NN	O	O
obliterans	NN	O	O
after	NN	O	O
transplantation	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
whether	NN	O	O
CMV	NN	O	O
induces	NN	O	O
IL-6	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
CMV	NN	O	O
infection	NN	O	O
increased	NN	O	O
both	NN	O	O
IL-6	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
mRNA	NN	O	B-RNA
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
CMV	NN	O	B-protein
immediate	NN	O	I-protein
early	NN	O	I-protein
1	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
CMV	NN	O	B-protein
immediate	NN	O	I-protein
early	NN	O	I-protein
1	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
was	NN	O	O
dependent	NN	O	O
upon	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
specific	NN	O	O
transcription	NN	O	B-DNA
factor	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
CMV	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
cofactor	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
rejection	NN	O	O
and	NN	O	O
infection	NN	O	O
after	NN	O	O
transplantation	NN	O	O
through	NN	O	O
its	NN	O	O
effects	NN	O	O
on	NN	O	O
IL-6	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Vitamin	NN	O	O
D3-	NN	O	O
and	NN	O	O
retinoic	NN	O	O
acid-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
:	NN	O	O
interactions	NN	O	O
between	NN	O	O
the	NN	O	O
endogenous	NN	O	B-protein
vitamin	NN	O	I-protein
D3	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
and	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
alpha-dihydroxycholecalciferol	NN	O	O
(	NN	O	O
VitD3	NN	O	O
)	NN	O	O
are	NN	O	O
potent	NN	O	O
regulators	NN	O	O
of	NN	O	O
hematopoletic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Yet	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
as	NN	O	O
to	NN	O	O
how	NN	O	O
the	NN	O	O
RA	NN	O	B-protein
and	NN	O	I-protein
VitD3	NN	O	I-protein
receptor	NN	O	I-protein
network	NN	O	I-protein
operates	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
whether	NN	O	O
receptor	NN	O	O
interactions	NN	O	O
can	NN	O	O
explain	NN	O	O
the	NN	O	O
interplay	NN	O	O
between	NN	O	O
the	NN	O	O
RA	NN	O	O
-and	NN	O	O
VitD3-signaling	NN	O	O
pathways	NN	O	O
during	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
expression	NN	O	O
,	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-protein
RA	NN	O	I-protein
and	NN	O	I-protein
VitD3	NN	O	I-protein
receptors	NN	O	I-protein
[	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RARs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RXRs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
VitD3	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
]	NN	O	O
in	NN	O	O
the	NN	O	O
U-937	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
which	NN	O	O
RA	NN	O	O
and	NN	O	O
VitD3	NN	O	O
induce	NN	O	O
distinct	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
pathways	NN	O	O
.	NN	O	O

VitD3	NN	O	O
induction	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
VDR/RXR	NN	O	B-protein
DNA-binding	NN	O	I-protein
complexes	NN	O	I-protein
on	NN	O	O
both	NN	O	O
VitD3	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
and	NN	O	O
RA	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
(	NN	O	O
RAREs	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
was	NN	O	O
only	NN	O	O
observed	NN	O	O
from	NN	O	O
a	NN	O	O
VitD3	NN	O	B-DNA
response	NN	O	I-DNA
element-driven	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
.	NN	O	O

Several	NN	O	O
DNA-binding	NN	O	B-protein
complexes	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
on	NN	O	O
RAREs	NN	O	B-DNA
in	NN	O	O
undifferentiated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Stimulation	NN	O	O
by	NN	O	O
RA	NN	O	O
resulted	NN	O	O
in	NN	O	O
increased	NN	O	O
RAR	NN	O	B-protein
beta	NN	O	I-protein
/RXR	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
activated	NN	O	O
RARE	NN	O	B-DNA
-dependent	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
RAR-beta	NN	O	B-protein
.	NN	O	O

Concomitant	NN	O	O
stimulation	NN	O	O
by	NN	O	O
VitD3	NN	O	O
inhibited	NN	O	O
the	NN	O	O
RA-stimulated	NN	O	O
formation	NN	O	O
of	NN	O	O
RAR	NN	O	B-protein
beta/RXR	NN	O	I-protein
heterodimers	NN	O	I-protein
,	NN	O	O
favoring	NN	O	O
VDR	NN	O	B-protein
/RXR	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
RARE	NN	O	B-DNA
.	NN	O	O

Also	NN	O	O
,	NN	O	O
VitD3	NN	O	O
inhibited	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CD23	NN	O	B-protein
and	NN	O	O
CD49f	NN	O	B-protein
,	NN	O	O
characteristic	NN	O	O
markers	NN	O	O
of	NN	O	O
retinoid-induced	NN	O	O
U-937	NN	O	B-cell_line
cell	NN	O	I-cell_line
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
neither	NN	O	O
the	NN	O	O
RA-stimulated	NN	O	O
,	NN	O	O
RARE	NN	O	B-DNA
-mediated	NN	O	O
transcription	NN	O	O
nor	NN	O	O
the	NN	O	O
induced	NN	O	O
RAR-beta	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
suppressed	NN	O	O
by	NN	O	O
VitD3	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
VitD3	NN	O	O
selectively	NN	O	O
inhibited	NN	O	O
the	NN	O	O
retinoid-induced	NN	O	O
differentiation	NN	O	O
program	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
RARE	NN	O	B-DNA
-mediated	NN	O	O
signal	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
complex	NN	O	O
role	NN	O	O
for	NN	O	O
VitD3	NN	O	O
in	NN	O	O
modifying	NN	O	O
the	NN	O	O
retinoid	NN	O	O
differentiation	NN	O	O
pathway	NN	O	O
and	NN	O	O
may	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
differentiation-inducing	NN	O	O
therapy	NN	O	O
of	NN	O	O
hematopoietic	NN	O	O
tumors	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
posttranscriptional	NN	O	I-protein
trans-activator	NN	O	I-protein
Rex	NN	O	B-protein
contains	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
export	NN	O	I-protein
signal	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Rex	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
nuclear	NN	O	O
export	NN	O	O
of	NN	O	O
unspliced	NN	O	B-RNA
viral	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
for	NN	O	O
virus	NN	O	O
replication	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
manuscript	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Rex	NN	O	B-protein
shuttles	NN	O	O
between	NN	O	O
the	NN	O	O
nucleus	NN	O	O
and	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
and	NN	O	O
that	NN	O	O
its	NN	O	O
activation	NN	O	B-protein
domain	NN	O	I-protein
constitutes	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
export	NN	O	I-protein
signal	NN	O	I-protein
that	NN	O	O
specifies	NN	O	O
efficient	NN	O	O
transport	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
Rex	NN	O	B-protein
-mediated	NN	O	O
trans-activation	NN	O	O
in	NN	O	O
which	NN	O	O
Rex-viral	NN	O	B-protein
mRNA	NN	O	I-protein
complexes	NN	O	I-protein
are	NN	O	O
targeted	NN	O	O
for	NN	O	O
nuclear	NN	O	O
export	NN	O	O
by	NN	O	O
the	NN	O	O
direct	NN	O	O
action	NN	O	O
of	NN	O	O
the	NN	O	O
activation	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
preferentially	NN	O	O
activates	NN	O	O
the	NN	O	O
94-kD	NN	O	B-protein
STAT5A	NN	O	I-protein
and	NN	O	O
an	NN	O	O
80-kD	NN	O	B-protein
STAT5A	NN	O	I-protein
isoform	NN	O	I-protein
in	NN	O	O
human	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
monocytes	NN	O	O
.	NN	O	O

Granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
induces	NN	O	O
immediate	NN	O	O
effects	NN	O	O
in	NN	O	O
monocytes	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
JAK2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
STAT	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
STAT5	NN	O	B-protein
)	NN	O	O
pathway	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
identified	NN	O	O
homologues	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
STAT5A	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT5B	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
lower	NN	O	O
molecular	NN	O	O
weight	NN	O	O
variants	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
.	NN	O	O

To	NN	O	O
define	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
STAT5	NN	O	B-protein
homologues	NN	O	I-protein
and	NN	O	O
lower	NN	O	O
molecular	NN	O	O
weight	NN	O	O
variant	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
differentiated	NN	O	O
into	NN	O	O
macrophages	NN	O	B-cell_type
by	NN	O	O
culture	NN	O	O
in	NN	O	O
macrophage-CSF	NN	O	B-protein
(	NN	O	O
M-CSF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
measured	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-protein
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT5A	NN	O	B-protein
,	NN	O	O
STAT5B	NN	O	B-protein
,	NN	O	O
and	NN	O	O
any	NN	O	O
lower	NN	O	O
molecular	NN	O	O
weight	NN	O	O
STAT5	NN	O	B-protein
isoforms	NN	O	I-protein
.	NN	O	O

Freshly	NN	O	O
isolated	NN	O	O
monocytes	NN	O	O
expressed	NN	O	O
94-kD	NN	O	B-protein
STAT5A	NN	O	I-protein
,	NN	O	O
92-kD	NN	O	O
STAT5B	NN	O	B-protein
,	NN	O	O
and	NN	O	O
an	NN	O	O
80-kD	NN	O	B-protein
STAT5A	NN	O	I-protein
molecule	NN	O	I-protein
.	NN	O	O

Whereas	NN	O	O
94-kD	NN	O	B-protein
STAT5A	NN	O	I-protein
was	NN	O	O
clearly	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
response	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
GRR	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
of	NN	O	O
the	NN	O	O
Fc	NN	O	B-DNA
gamma	NN	O	I-DNA
RI	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
substantially	NN	O	O
less	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
STAT5B	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
immobilized	NN	O	O
GRR	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

Macrophages	NN	O	O
lost	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
express	NN	O	O
the	NN	O	O
80-kD	NN	O	B-protein
STAT5A	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
but	NN	O	O
retained	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
activate	NN	O	O
STAT5A	NN	O	B-protein
.	NN	O	O

STAT5A-STAT5A	NN	O	B-protein
homodimers	NN	O	I-protein
and	NN	O	O
STAT5A-STAT5B	NN	O	B-protein
heterodimers	NN	O	I-protein
formed	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT5A	NN	O	B-protein
predominates	NN	O	O
compared	NN	O	O
to	NN	O	O
STAT5B	NN	O	B-protein
when	NN	O	O
assayed	NN	O	O
by	NN	O	O
direct	NN	O	O
immunoprecipitation	NN	O	O
and	NN	O	O
by	NN	O	O
evaluation	NN	O	O
of	NN	O	O
bound	NN	O	O
STATs	NN	O	B-protein
to	NN	O	O
immobilized	NN	O	O
GRR	NN	O	B-DNA
.	NN	O	O

Selective	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
homologues	NN	O	I-protein
in	NN	O	O
addition	NN	O	O
to	NN	O	O
generation	NN	O	O
of	NN	O	O
lower	NN	O	O
molecular	NN	O	O
isoforms	NN	O	O
may	NN	O	O
provide	NN	O	O
specificity	NN	O	O
and	NN	O	O
control	NN	O	O
to	NN	O	O
genes	NN	O	O
expressed	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
cytokines	NN	O	B-protein
such	NN	O	O
as	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Translocation	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
14	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q27	NN	O	I-DNA
;	NN	O	I-DNA
q11	NN	O	I-DNA
)	NN	O	I-DNA
:	NN	O	O
a	NN	O	O
new	NN	O	O
variant	NN	O	O
translocation	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
of	NN	O	O
B-cell	NN	O	O
type	NN	O	O
with	NN	O	O
BCL6	NN	O	B-DNA
rearrangement	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
a	NN	O	O
65-year-old	NN	O	O
woman	NN	O	O
with	NN	O	O
non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
NHL	NN	O	O
)	NN	O	O
carrying	NN	O	O
a	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
14	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q27	NN	O	I-DNA
;	NN	O	I-DNA
q11	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
BCL6	NN	O	B-DNA
rearrangement	NN	O	O
in	NN	O	O
the	NN	O	O
affected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

She	NN	O	O
had	NN	O	O
generalized	NN	O	O
lymphadenopathy	NN	O	O
and	NN	O	O
the	NN	O	O
bone	NN	O	O
marrow	NN	O	O
was	NN	O	O
infiltrated	NN	O	O
by	NN	O	O
lymphoma	NN	O	B-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
presentation	NN	O	O
.	NN	O	O

Histological	NN	O	O
diagnosis	NN	O	O
was	NN	O	O
``	NN	O	O
malignant	NN	O	O
lymphoma	NN	O	O
,	NN	O	O
diffuse	NN	O	O
,	NN	O	O
large	NN	O	O
cell	NN	O	O
''	NN	O	O
type	NN	O	O
according	NN	O	O
to	NN	O	O
an	NN	O	O
International	NN	O	O
Working	NN	O	O
Formulation	NN	O	O
.	NN	O	O

Chromosome	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
a	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
14	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q27	NN	O	I-DNA
;	NN	O	I-DNA
q11	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
new	NN	O	O
variant	NN	O	O
translocation	NN	O	O
of	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
14	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q27	NN	O	I-DNA
;	NN	O	I-DNA
q32	NN	O	I-DNA
)	NN	O	I-DNA
.	NN	O	O

Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
BCL6	NN	O	B-DNA
,	NN	O	O
JH	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
TCR	NN	O	B-DNA
beta	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
of	NN	O	O
TCR	NN	O	B-DNA
delta	NN	O	I-DNA
.	NN	O	O

Cosmid	NN	O	B-DNA
probe	NN	O	I-DNA
of	NN	O	O
BCL6	NN	O	B-DNA
hybridized	NN	O	O
to	NN	O	O
14q11	NN	O	B-DNA
and	NN	O	O
3q27	NN	O	B-DNA
by	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
band	NN	O	B-DNA
14q11	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
locus	NN	O	O
of	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
alpha-	NN	O	I-protein
and	NN	O	I-protein
delta-chains	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
alpha/delta	NN	O	I-protein
)	NN	O	O
,	NN	O	O
lymphoma	NN	O	B-cell_type
cells	NN	O	I-cell_type
expressed	NN	O	O
B-cell	NN	O	B-cell_type
,	NN	O	I-cell_type
IgGk	NN	O	I-cell_type
phenotype	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
novel	NN	O	B-DNA
proto-oncogene	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
vicinity	NN	O	O
of	NN	O	O
TCR	NN	O	B-DNA
alpha/delta	NN	O	I-DNA
is	NN	O	O
involved	NN	O	O
in	NN	O	O
this	NN	O	O
translocation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transgenic	NN	O	O
expression	NN	O	O
of	NN	O	O
PML/RARalpha	NN	O	B-protein
impairs	NN	O	O
myelopoiesis	NN	O	O
.	NN	O	O

The	NN	O	O
translocation	NN	O	O
found	NN	O	O
in	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
rearranges	NN	O	O
the	NN	O	O
promyelocytic	NN	O	B-DNA
leukemia	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
PML	NN	O	B-DNA
)	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
15	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
retinoic	NN	O	B-DNA
acid	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	O
RARalpha	NN	O	B-protein
)	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
17	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
yields	NN	O	O
a	NN	O	O
fusion	NN	O	O
transcript	NN	O	O
,	NN	O	O
PML/RARalpha	NN	O	B-protein
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
with	NN	O	O
reported	NN	O	O
dominant	NN	O	O
negative	NN	O	O
functions	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
hormone	NN	O	O
.	NN	O	O

Clinical	NN	O	O
remissions	NN	O	O
induced	NN	O	O
with	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
treatment	NN	O	O
in	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
are	NN	O	O
linked	NN	O	O
to	NN	O	O
PML/RARalpha	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
the	NN	O	O
PML/RARalpha	NN	O	B-protein
role	NN	O	O
in	NN	O	O
myelopoiesis	NN	O	O
,	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
expressing	NN	O	O
PML/RARalpha	NN	O	B-protein
were	NN	O	O
engineered	NN	O	O
.	NN	O	O

A	NN	O	O
full-length	NN	O	O
PML/RARalpha	NN	O	B-DNA
cDNA	NN	O	I-DNA
driven	NN	O	O
by	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Expression	NN	O	O
was	NN	O	O
confirmed	NN	O	O
in	NN	O	O
the	NN	O	O
bone	NN	O	O
marrow	NN	O	O
with	NN	O	O
a	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
PCR	NN	O	O
assay	NN	O	O
.	NN	O	O

Basal	NN	O	O
total	NN	O	O
white	NN	O	O
blood	NN	O	O
cell	NN	O	O
and	NN	O	O
granulocyte	NN	O	O
counts	NN	O	O
did	NN	O	O
not	NN	O	O
appreciably	NN	O	O
differ	NN	O	O
between	NN	O	O
PML/RARalpha	NN	O	B-protein
transgenic	NN	O	O
and	NN	O	O
control	NN	O	O
mice	NN	O	O
.	NN	O	O

Cell	NN	O	O
sorter	NN	O	O
analysis	NN	O	O
of	NN	O	O
CD11b+	NN	O	B-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
revealed	NN	O	O
similar	NN	O	O
CD11b+	NN	O	B-cell_type
populations	NN	O	I-cell_type
in	NN	O	O
transgenic	NN	O	O
and	NN	O	O
control	NN	O	O
mice	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
clonal	NN	O	O
growth	NN	O	O
assays	NN	O	O
performed	NN	O	O
on	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
from	NN	O	O
transgenic	NN	O	O
versus	NN	O	O
control	NN	O	O
mice	NN	O	O
revealed	NN	O	O
a	NN	O	O
marked	NN	O	O
reduction	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
,	NN	O	O
especially	NN	O	O
in	NN	O	O
those	NN	O	O
responding	NN	O	O
to	NN	O	O
granulocyte/	NN	O	B-protein
macrophage	NN	O	I-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Granulocyte/macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
kit	NN	O	O
ligand	NN	O	O
cotreatment	NN	O	O
did	NN	O	O
not	NN	O	O
overcome	NN	O	O
this	NN	O	O
inhibition	NN	O	O
.	NN	O	O

Impaired	NN	O	O
myelopoiesis	NN	O	O
in	NN	O	O
vivo	NN	O	O
was	NN	O	O
shown	NN	O	O
by	NN	O	O
stressing	NN	O	O
these	NN	O	O
mice	NN	O	O
with	NN	O	O
sublethal	NN	O	O
irradiation	NN	O	O
.	NN	O	O

Following	NN	O	O
irradiation	NN	O	O
,	NN	O	O
PML/RARalpha	NN	O	B-protein
transgenic	NN	O	O
mice	NN	O	O
,	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
controls	NN	O	O
,	NN	O	O
more	NN	O	O
rapidly	NN	O	O
depressed	NN	O	O
peripheral	NN	O	O
white	NN	O	O
blood	NN	O	O
cell	NN	O	O
and	NN	O	O
granulocyte	NN	O	O
counts	NN	O	O
.	NN	O	O

As	NN	O	O
expected	NN	O	O
,	NN	O	O
nearly	NN	O	O
all	NN	O	O
control	NN	O	O
mice	NN	O	O
(	NN	O	O
94.4	NN	O	O
%	NN	O	O
)	NN	O	O
survived	NN	O	O
irradiation	NN	O	O
,	NN	O	O
yet	NN	O	O
this	NN	O	O
irradiation	NN	O	O
was	NN	O	O
lethal	NN	O	O
to	NN	O	O
45.8	NN	O	O
%	NN	O	O
of	NN	O	O
PML/RARalpha	NN	O	B-protein
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Lethality	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
more	NN	O	O
severe	NN	O	O
leukopenia	NN	O	O
in	NN	O	O
transgenic	NN	O	O
versus	NN	O	O
control	NN	O	O
mice	NN	O	O
.	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
treatment	NN	O	O
of	NN	O	O
irradiated	NN	O	O
PML/RARalpha	NN	O	B-protein
mice	NN	O	O
enhanced	NN	O	O
granulocyte	NN	O	B-cell_type
recovery	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
abnormal	NN	O	O
myelopoiesis	NN	O	O
due	NN	O	O
to	NN	O	O
PML/RARalpha	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
an	NN	O	O
early	NN	O	O
event	NN	O	O
in	NN	O	O
oncogenic	NN	O	O
transformation	NN	O	O
.	NN	O	O

-DOCSTART-	O

C/EBP	NN	O	B-protein
activators	NN	O	I-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
and	NN	O	O
proviral	NN	O	O
induction	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Previous	NN	O	O
work	NN	O	O
has	NN	O	O
shown	NN	O	O
that	NN	O	O
C/EBP	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
C/EBP	NN	O	B-protein
transcriptional	NN	O	I-protein
activators	NN	O	I-protein
are	NN	O	O
necessary	NN	O	O
for	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
that	NN	O	O
C/EBP	NN	O	B-protein
proteins	NN	O	I-protein
play	NN	O	O
in	NN	O	O
induction	NN	O	O
and	NN	O	O
replication	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

Ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
dominant	NN	O	O
negative	NN	O	O
C/EBP	NN	O	B-protein
protein	NN	O	I-protein
LIP	NN	O	I-protein
inhibited	NN	O	O
HIV-1	NN	O	O
mRNA	NN	O	O
and	NN	O	O
virus	NN	O	O
production	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_line
U1	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
C/EBP	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
provirus	NN	O	O
induction	NN	O	O
.	NN	O	O

U1	NN	O	B-cell_line
lines	NN	O	I-cell_line
overexpressing	NN	O	O
C/EBP	NN	O	B-protein
activator	NN	O	I-protein
NF-IL-6	NN	O	B-protein
produced	NN	O	O
more	NN	O	O
viral	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
virus	NN	O	O
particles	NN	O	O
following	NN	O	O
cellular	NN	O	O
activation	NN	O	O
than	NN	O	O
control	NN	O	B-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
C/EBP	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
limiting	NN	O	O
for	NN	O	O
virus	NN	O	O
transcription	NN	O	O
.	NN	O	O

HIV-1	NN	O	O
harboring	NN	O	O
mutations	NN	O	O
within	NN	O	O
two	NN	O	O
C/EBP	NN	O	B-DNA
sites	NN	O	I-DNA
were	NN	O	O
crippled	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
replicate	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
these	NN	O	O
sites	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
replication	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
identify	NN	O	O
C/EBP	NN	O	B-protein
proteins	NN	O	I-protein
as	NN	O	O
regulators	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
2	NN	O	I-protein
and	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
independently	NN	O	O
transactivate	NN	O	O
p53	NN	O	B-protein
through	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

B-cell	NN	O	B-cell_type
immortalization	NN	O	O
by	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
permanent	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	O
processes	NN	O	O
which	NN	O	O
normally	NN	O	O
regulate	NN	O	O
cell	NN	O	O
division	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
functions	NN	O	O
possessed	NN	O	O
by	NN	O	O
p53	NN	O	B-protein
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
normal	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

In	NN	O	O
studies	NN	O	O
initiated	NN	O	O
to	NN	O	O
evaluate	NN	O	O
relationships	NN	O	O
between	NN	O	O
EBV	NN	O	B-DNA
latent	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
p53	NN	O	B-protein
,	NN	O	O
p53	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
increase	NN	O	O
approximately	NN	O	O
10-fold	NN	O	O
4	NN	O	O
to	NN	O	O
5	NN	O	O
days	NN	O	O
after	NN	O	O
EBV	NN	O	O
infection	NN	O	O
of	NN	O	O
purified	NN	O	O
resting	NN	O	B-cell_type
human	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
the	NN	O	O
induced	NN	O	O
p53	NN	O	B-protein
was	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
.	NN	O	O

Latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
EBV	NN	O	B-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
2	NN	O	I-protein
mediated	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
p53	NN	O	B-protein
levels	NN	O	O
via	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Cooperation	NN	O	O
between	NN	O	O
core	NN	O	B-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
adjacent	NN	O	B-DNA
promoter	NN	O	I-DNA
elements	NN	O	I-DNA
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
interleukin-3	NN	O	B-protein
.	NN	O	O

Tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
interleukin-3	NN	O	B-protein
(	NN	O	O
IL-3	NN	O	B-protein
)	NN	O	O
is	NN	O	O
mediated	NN	O	O
via	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
located	NN	O	O
within	NN	O	O
315	NN	O	O
base	NN	O	O
pairs	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
is	NN	O	O
achieved	NN	O	O
in	NN	O	O
part	NN	O	O
through	NN	O	O
the	NN	O	O
positive	NN	O	O
activities	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
and	NN	O	I-DNA
Elf-1	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
contribution	NN	O	O
to	NN	O	O
T	NN	O	O
cell-specific	NN	O	O
expression	NN	O	O
by	NN	O	O
other	NN	O	O
promoter	NN	O	B-DNA
sites	NN	O	I-DNA
was	NN	O	O
assessed	NN	O	O
in	NN	O	O
a	NN	O	O
transient	NN	O	O
expression	NN	O	O
assay	NN	O	O
with	NN	O	O
IL-3	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
a	NN	O	O
luciferase	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
focusing	NN	O	O
initially	NN	O	O
on	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
binding	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
CBF	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
footprinted	NN	O	O
in	NN	O	O
vivo	NN	O	O
upon	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Activity	NN	O	O
of	NN	O	O
the	NN	O	O
CBF	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
critically	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
adjacent	NN	O	B-DNA
activator	NN	O	I-DNA
site	NN	O	I-DNA
Act-1	NN	O	I-DNA
.	NN	O	O

Together	NN	O	O
the	NN	O	O
Act-1	NN	O	B-DNA
and	NN	O	O
CBF	NN	O	B-DNA
sites	NN	O	I-DNA
form	NN	O	O
a	NN	O	O
functional	NN	O	B-DNA
unit	NN	O	I-DNA
(	NN	O	O
AC	NN	O	B-DNA
unit	NN	O	I-DNA
)	NN	O	O
with	NN	O	O
dual	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
AC	NN	O	B-DNA
unit	NN	O	I-DNA
is	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
enhance	NN	O	O
basal	NN	O	O
activity	NN	O	O
of	NN	O	O
promoters	NN	O	O
both	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
activity	NN	O	O
is	NN	O	O
further	NN	O	O
inducible	NN	O	O
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
already	NN	O	O
identified	NN	O	O
NIP	NN	O	B-DNA
repressor	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
evidence	NN	O	O
is	NN	O	O
presented	NN	O	O
for	NN	O	O
a	NN	O	O
second	NN	O	B-DNA
repressor	NN	O	I-DNA
region	NN	O	I-DNA
that	NN	O	O
restricts	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
a	NN	O	O
novel	NN	O	O
positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
in	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
promoter	NN	O	I-DNA
between	NN	O	O
nucleotide	NN	O	O
-180	NN	O	O
and	NN	O	O
-210	NN	O	O
that	NN	O	O
leads	NN	O	O
to	NN	O	O
increased	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Together	NN	O	O
these	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
T	NN	O	O
cell	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-3	NN	O	B-protein
is	NN	O	O
not	NN	O	O
specified	NN	O	O
by	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
tissue-specific	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
instead	NN	O	O
involves	NN	O	O
multiple	NN	O	O
interacting	NN	O	B-DNA
elements	NN	O	I-DNA
that	NN	O	O
provide	NN	O	O
both	NN	O	O
specific	NN	O	O
positive	NN	O	O
regulation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
specific	NN	O	O
negative	NN	O	O
regulation	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Modulation	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
receptor	NN	O	I-protein
beta-chain	NN	O	I-protein
controls	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

IFN-gamma	NN	O	B-protein
has	NN	O	O
potent	NN	O	O
antiproliferative	NN	O	O
and	NN	O	O
apoptotic	NN	O	O
effects	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
are	NN	O	O
important	NN	O	O
in	NN	O	O
determining	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
and	NN	O	O
polarized	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
any	NN	O	O
event	NN	O	O
that	NN	O	O
enables	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
become	NN	O	O
less	NN	O	O
responsive	NN	O	O
to	NN	O	O
IFN-	NN	O	B-protein
gamma	NN	O	I-protein
may	NN	O	O
potentially	NN	O	O
alter	NN	O	O
immune	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
Ag	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
work	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
are	NN	O	O
stimulated	NN	O	O
through	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
expanded	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
are	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
a	NN	O	O
lack	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
jak	NN	O	B-protein
kinases	NN	O	I-protein
and	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
STAT1	NN	O	B-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
a	NN	O	O
signal	NN	O	O
transducer	NN	O	O
and	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

This	NN	O	O
nonresponsiveness	NN	O	O
occurs	NN	O	O
because	NN	O	O
of	NN	O	O
a	NN	O	O
lack	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
beta-	NN	O	B-protein
chain	NN	O	I-protein
(	NN	O	O
accessory	NN	O	O
factor	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
while	NN	O	O
at	NN	O	O
the	NN	O	O
same	NN	O	O
time	NN	O	O
maintaining	NN	O	O
IFN-gamma	NN	O	B-protein
receptor	NN	O	I-protein
alpha-chain	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
beta-chain	NN	O	B-protein
can	NN	O	O
be	NN	O	O
restored	NN	O	O
by	NN	O	O
secondary	NN	O	O
TCR	NN	O	B-protein
ligation	NN	O	O
or	NN	O	O
PMA	NN	O	O
treatment	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cell	NN	O	I-cell_type
blasts	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
PMA	NN	O	O
are	NN	O	O
now	NN	O	O
responsive	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

When	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
,	NN	O	O
highly	NN	O	O
enriched	NN	O	O
(	NN	O	O
>	NN	O	O
98	NN	O	O
%	NN	O	O
)	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
examined	NN	O	O
for	NN	O	O
IFN-gamma	NN	O	B-protein
responsiveness	NN	O	O
;	NN	O	O
these	NN	O	O
cells	NN	O	O
can	NN	O	O
respond	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
express	NN	O	O
beta-chain	NN	O	B-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
as	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
progress	NN	O	O
from	NN	O	O
primary	NN	O	O
TCR	NN	O	B-protein
activation	NN	O	O
through	NN	O	O
IL-2	NN	O	B-protein
-dependent	NN	O	O
proliferation	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
secondary	NN	O	O
TCR	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
their	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
varies	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
may	NN	O	O
affect	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
participate	NN	O	O
in	NN	O	O
an	NN	O	O
ongoing	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Age-related	NN	O	O
decreases	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
associated	NN	O	O
with	NN	O	O
impaired	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
transcriptional	NN	O	I-protein
factors	NN	O	I-protein
AP-1	NN	O	B-protein
and	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
transcriptional	NN	O	B-protein
factors	NN	O	I-protein
AP-1	NN	O	B-protein
and	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
of	NN	O	O
activated	NN	O	O
T	NN	O	O
cells	NN	O	O
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
normal	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
it	NN	O	O
is	NN	O	O
unknown	NN	O	O
if	NN	O	O
the	NN	O	O
age-related	NN	O	O
decline	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
by	NN	O	O
activated	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
aberrancies	NN	O	O
in	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
current	NN	O	O
studies	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
elderly	NN	O	O
(	NN	O	O
mean	NN	O	O
78	NN	O	O
years	NN	O	O
)	NN	O	O
and	NN	O	O
young	NN	O	O
(	NN	O	O
mean	NN	O	O
37	NN	O	O
years	NN	O	O
)	NN	O	O
humans	NN	O	O
was	NN	O	O
measured	NN	O	O
in	NN	O	O
cultures	NN	O	O
stimulated	NN	O	O
with	NN	O	O
PHA	NN	O	B-protein
,	NN	O	O
PHA	NN	O	B-protein
plus	NN	O	O
PMA	NN	O	O
,	NN	O	O
crosslinked	NN	O	O
anti-CD3	NN	O	O
mAB	NN	O	O
OKT3	NN	O	O
plus	NN	O	O
PMA	NN	O	O
,	NN	O	O
or	NN	O	O
PMA	NN	O	O
plus	NN	O	O
ionomycin	NN	O	O
.	NN	O	O

Substantial	NN	O	O
decreases	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
were	NN	O	O
observed	NN	O	O
for	NN	O	O
cell	NN	O	O
cultures	NN	O	O
from	NN	O	O
7	NN	O	O
of	NN	O	O
12	NN	O	O
elderly	NN	O	O
individuals	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
different	NN	O	O
stimuli	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
produced	NN	O	O
by	NN	O	O
stimulated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
other	NN	O	O
elderly	NN	O	O
individuals	NN	O	O
were	NN	O	O
equivalent	NN	O	O
to	NN	O	O
those	NN	O	O
observed	NN	O	O
for	NN	O	O
stimulated	NN	O	O
T	NN	O	O
cells	NN	O	O
of	NN	O	O
young	NN	O	O
subjects	NN	O	O
.	NN	O	O

Analyses	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
DNA	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
decreased	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
by	NN	O	O
stimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
elderly	NN	O	O
individuals	NN	O	O
was	NN	O	O
closely	NN	O	O
associated	NN	O	O
with	NN	O	O
impairments	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
elderly	NN	O	O
subjects	NN	O	O
with	NN	O	O
normal	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
exhibited	NN	O	O
normal	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
competition	NN	O	O
experiments	NN	O	O
analyzing	NN	O	O
the	NN	O	O
normal	NN	O	O
components	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
showed	NN	O	O
that	NN	O	O
the	NN	O	O
age-related	NN	O	O
reductions	NN	O	O
in	NN	O	O
stimulus-dependent	NN	O	O
NF-AT	NN	O	B-protein
complexes	NN	O	I-protein
corresponded	NN	O	O
to	NN	O	O
the	NN	O	O
slow	NN	O	O
migrating	NN	O	O
complexes	NN	O	O
that	NN	O	O
were	NN	O	O
composed	NN	O	O
of	NN	O	O
c-Fos/c-Jun	NN	O	B-protein
AP-1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
resting	NN	O	O
and	NN	O	O
stimulated	NN	O	O
levels	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
were	NN	O	O
reduced	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
certain	NN	O	O
elderly	NN	O	O
individuals	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
alterations	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
did	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
changes	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
age-related	NN	O	O
impairments	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-AT	NN	O	B-protein
are	NN	O	O
closely	NN	O	O
associated	NN	O	O
with	NN	O	O
decreased	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
further	NN	O	O
suggest	NN	O	O
that	NN	O	O
aberrancies	NN	O	O
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
c-Fos/c-Jun	NN	O	B-protein
AP-1	NN	O	B-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
impaired	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

IL-13	NN	O	B-protein
induces	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
JAK2	NN	O	B-protein
Janus	NN	O	I-protein
kinase	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
colon	NN	O	I-cell_line
carcinoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
:	NN	O	O
similarities	NN	O	O
between	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
reported	NN	O	O
that	NN	O	O
IL-13R	NN	O	B-protein
may	NN	O	O
share	NN	O	O
a	NN	O	O
component	NN	O	O
with	NN	O	O
IL-4R	NN	O	B-protein
.	NN	O	O
Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
both	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
share	NN	O	O
signaling	NN	O	O
events	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
colon	NN	O	I-cell_line
carcinoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
HT-29	NN	O	B-cell_line
and	NN	O	O
WiDr	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

IL-13	NN	O	B-protein
caused	NN	O	O
rapid	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
out	NN	O	O
of	NN	O	O
four	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
known	NN	O	O
Janus	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
kinases	NN	O	I-protein
(	NN	O	O
JAKs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
JAK2	NN	O	B-protein
kinase	NN	O	I-protein
is	NN	O	O
rapidly	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-13	NN	O	B-protein
.	NN	O	O

Within	NN	O	O
1	NN	O	O
min	NN	O	O
of	NN	O	O
activation	NN	O	O
,	NN	O	O
JAK2	NN	O	B-protein
was	NN	O	O
phosphorylated	NN	O	O
,	NN	O	O
and	NN	O	O
peaked	NN	O	O
in	NN	O	O
10	NN	O	O
min	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
IL-13	NN	O	B-protein
phosphorylated	NN	O	O
insulin	NN	O	B-protein
response	NN	O	I-protein
substrate-1	NN	O	I-protein
,	NN	O	O
IL-4R	NN	O	B-protein
p140	NN	O	I-protein
,	NN	O	O
JAK1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Tyk2	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
JAK3	NN	O	B-protein
kinase	NN	O	I-protein
.	NN	O	O

IL-4	NN	O	B-protein
also	NN	O	O
stimulated	NN	O	O
all	NN	O	O
three	NN	O	O
kinases	NN	O	B-protein
and	NN	O	O
substrates	NN	O	O
,	NN	O	O
but	NN	O	O
unlike	NN	O	O
in	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
IL-4	NN	O	B-protein
did	NN	O	O
not	NN	O	O
involve	NN	O	O
JAK3	NN	O	B-protein
activation	NN	O	O
for	NN	O	O
its	NN	O	O
signaling	NN	O	O
in	NN	O	O
colon	NN	O	B-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
JAK2	NN	O	B-protein
associated	NN	O	O
with	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
p140	NN	O	I-protein
before	NN	O	O
and	NN	O	O
after	NN	O	O
stimulation	NN	O	O
with	NN	O	O
IL-13	NN	O	B-protein
.	NN	O	O

Both	NN	O	O
IL-13	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
STAT	NN	O	I-protein
(	NN	O	O
STAT6	NN	O	B-protein
)	NN	O	O
but	NN	O	O
not	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
or	NN	O	O
STAT5	NN	O	B-protein
.	NN	O	O

125I-IL-13	NN	O	B-protein
did	NN	O	O
not	NN	O	O
bind	NN	O	O
to	NN	O	O
colon	NN	O	B-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
unlabeled	NN	O	O
IL-13	NN	O	B-protein
competed	NN	O	O
for	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
125I-IL-4	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
IL-13	NN	O	B-protein
utilizes	NN	O	O
IL-4R	NN	O	B-protein
and	NN	O	O
its	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
,	NN	O	O
and	NN	O	O
JAK2	NN	O	B-protein
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
IL-4R	NN	O	B-protein
and	NN	O	O
IL-13R	NN	O	B-protein
in	NN	O	O
colon	NN	O	B-cell_type
cancer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-6	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
and	NN	O	O
expression	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

IL-4	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
inhibit	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
and	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
by	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
transcriptional	NN	O	O
or	NN	O	O
post-transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-6	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
we	NN	O	O
studied	NN	O	O
IL-6	NN	O	B-protein
production	NN	O	O
,	NN	O	O
expression	NN	O	O
level	NN	O	O
of	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
IL-6	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
NF-IL-6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
stability	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
and	NN	O	O
U937	NN	O	B-cell_line
,	NN	O	O
stimulated	NN	O	O
by	NN	O	O
PMA	NN	O	O
and	NN	O	O
LPS	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
or	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
or	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

Both	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
were	NN	O	O
seen	NN	O	O
to	NN	O	O
inhibit	NN	O	O
IL-6	NN	O	B-protein
production	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
both	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
studied	NN	O	O
.	NN	O	O

In	NN	O	O
chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
assays	NN	O	O
,	NN	O	O
utilizing	NN	O	O
the	NN	O	O
transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
a	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
reporter	NN	O	I-DNA
plasmid	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
suppressed	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	O
gene	NN	O	O
promoter	NN	O	O
stimulated	NN	O	O
by	NN	O	O
PMA	NN	O	O
and	NN	O	O
LPS	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
inhibited	NN	O	O
nuclear	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
NF-	NN	O	O
IL-6	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
enhanced	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
a	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
assay	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
may	NN	O	O
inhibit	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
affecting	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
while	NN	O	O
IL-10	NN	O	B-protein
may	NN	O	O
inhibit	NN	O	O
the	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
post-transcriptionally	NN	O	O
,	NN	O	O
without	NN	O	O
suppressing	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
inhibit	NN	O	O
IL-6	NN	O	B-protein
production	NN	O	O
by	NN	O	O
different	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
in	NN	O	O
a	NN	O	O
cyclosporin	NN	O	O
A-resistant	NN	O	O
pathway	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
drug	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
is	NN	O	O
the	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
Ca2+/calmodulin-dependent	NN	O	B-protein
serine-threonine	NN	O	I-protein
phosphatase	NN	O	I-protein
calcineurin	NN	O	B-protein
by	NN	O	O
the	NN	O	O
drug-immunophilin	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

Inactive	NN	O	O
calcineurin	NN	O	B-protein
is	NN	O	O
unable	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
required	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

IL-2	NN	O	B-protein
production	NN	O	O
by	NN	O	O
CsA-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
therefore	NN	O	O
dramatically	NN	O	O
reduced	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
that	NN	O	O
NFAT	NN	O	B-protein
can	NN	O	O
be	NN	O	O
activated	NN	O	O
,	NN	O	O
and	NN	O	O
significant	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
can	NN	O	O
be	NN	O	O
produced	NN	O	O
by	NN	O	O
the	NN	O	O
CsA-resistant	NN	O	O
CD28	NN	O	B-protein
-signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

In	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
,	NN	O	O
both	NN	O	O
multicopy	NN	O	O
NFAT	NN	O	B-protein
-and	NN	O	O
IL-2	NN	O	B-DNA
promoter-beta-galactosidase	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
could	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
/alpha-	NN	O	O
CD28	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
activation	NN	O	O
was	NN	O	O
resistant	NN	O	O
to	NN	O	O
CsA	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
showed	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
CsA-resistant	NN	O	B-protein
NFAT	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
PMA	NN	O	O
plus	NN	O	O
alphaCD28	NN	O	B-protein
.	NN	O	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
PMA	NN	O	O
/alphaCD28	NN	O	B-protein
produced	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
CsA	NN	O	O
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
NFAT	NN	O	B-protein
can	NN	O	O
be	NN	O	O
activated	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
can	NN	O	O
be	NN	O	O
produced	NN	O	O
in	NN	O	O
a	NN	O	O
calcineurin	NN	O	B-protein
independent	NN	O	O
manner	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
interleukin-10	NN	O	B-protein
on	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cell	NN	O	I-cell_type
responses	NN	O	O
to	NN	O	O
Cryptococcus	NN	O	O
neoformans	NN	O	O
,	NN	O	O
Candida	NN	O	O
albicans	NN	O	O
,	NN	O	O
and	NN	O	O
lipopolysaccharide	NN	O	O
.	NN	O	O

Deactivation	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
is	NN	O	O
critical	NN	O	O
to	NN	O	O
limit	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
but	NN	O	O
can	NN	O	O
be	NN	O	O
detrimental	NN	O	O
in	NN	O	O
the	NN	O	O
face	NN	O	O
of	NN	O	O
progressive	NN	O	O
infection	NN	O	O
.	NN	O	O

We	NN	O	O
compared	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
deactivating	NN	O	B-protein
cytokine	NN	O	I-protein
interleukin	NN	O	B-protein
10	NN	O	I-protein
(	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cell	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
responses	NN	O	O
to	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
Cryptococcus	NN	O	O
neoformans	NN	O	O
,	NN	O	O
and	NN	O	O
Candida	NN	O	O
albicans	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
effected	NN	O	O
dose-dependent	NN	O	O
inhibition	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
release	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
stimulated	NN	O	O
by	NN	O	O
LPS	NN	O	O
and	NN	O	O
C.	NN	O	O
neoformans	NN	O	O
,	NN	O	O
with	NN	O	O
significant	NN	O	O
inhibition	NN	O	O
seen	NN	O	O
with	NN	O	O
0.1	NN	O	O
U/ml	NN	O	O
and	NN	O	O
greater	NN	O	O
than	NN	O	O
90	NN	O	O
%	NN	O	O
inhibition	NN	O	O
noted	NN	O	O
with	NN	O	O
10	NN	O	O
U/ml	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
even	NN	O	O
at	NN	O	O
doses	NN	O	O
as	NN	O	O
high	NN	O	O
as	NN	O	O
100	NN	O	O
U/ml	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
inhibited	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
C.	NN	O	O
albicans	NN	O	O
by	NN	O	O
only	NN	O	O
50	NN	O	O
%	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
profoundly	NN	O	O
inhibited	NN	O	O
release	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
from	NN	O	O
PBMC	NN	O	B-cell_type
stimulated	NN	O	O
by	NN	O	O
all	NN	O	O
three	NN	O	O
stimuli	NN	O	O
.	NN	O	O

TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
release	NN	O	O
was	NN	O	O
inhibited	NN	O	O
even	NN	O	O
if	NN	O	O
IL-10	NN	O	B-protein
was	NN	O	O
added	NN	O	O
up	NN	O	O
to	NN	O	O
8	NN	O	O
h	NN	O	O
after	NN	O	O
cryptococcal	NN	O	O
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-1	NN	O	B-RNA
beta	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
of	NN	O	O
lesser	NN	O	O
magnitude	NN	O	O
and	NN	O	O
occurred	NN	O	O
only	NN	O	O
when	NN	O	O
IL-10	NN	O	B-protein
was	NN	O	O
added	NN	O	O
within	NN	O	O
2	NN	O	O
h	NN	O	O
of	NN	O	O
cryptococcal	NN	O	O
stimulation	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
inhibited	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
fungal	NN	O	O
stimuli	NN	O	O
.	NN	O	O

All	NN	O	O
three	NN	O	O
stimuli	NN	O	O
induced	NN	O	O
IL-10	NN	O	B-protein
production	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
,	NN	O	O
although	NN	O	O
over	NN	O	O
10-fold	NN	O	O
less	NN	O	O
IL-10	NN	O	B-protein
was	NN	O	O
released	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
C.	NN	O	O
neoformans	NN	O	O
compared	NN	O	O
with	NN	O	O
LPS	NN	O	O
and	NN	O	O
C.	NN	O	O
albicans	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
while	NN	O	O
IL-10	NN	O	B-protein
has	NN	O	O
deactivating	NN	O	O
effects	NN	O	O
on	NN	O	O
PBMC	NN	O	B-cell_type
responses	NN	O	O
to	NN	O	O
all	NN	O	O
three	NN	O	O
stimuli	NN	O	O
,	NN	O	O
disparate	NN	O	O
stimulus-	NN	O	O
and	NN	O	O
response-specific	NN	O	O
patterns	NN	O	O
of	NN	O	O
deactivation	NN	O	O
are	NN	O	O
seen	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
release	NN	O	O
appears	NN	O	O
to	NN	O	O
occur	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
for	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
both	NN	O	O
transcriptionally	NN	O	O
and	NN	O	O
posttranscriptionally	NN	O	O
for	NN	O	O
IL-1beta	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
and	NN	O	O
purification	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
substrate	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Identification	NN	O	O
as	NN	O	O
lymphocyte-specific	NN	O	B-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
LSP1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
B-chronic	NN	O	B-cell_line
lymphocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
(	NN	O	I-cell_line
B-CLL	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
two	NN	O	O
major	NN	O	O
PKC	NN	O	B-protein
substrates	NN	O	I-protein
,	NN	O	O
MARCKS	NN	O	B-protein
(	NN	O	O
myristoylated	NN	O	B-protein
,	NN	O	I-protein
alanine-rich	NN	O	I-protein
C	NN	O	I-protein
kinase	NN	O	I-protein
substrate	NN	O	I-protein
)	NN	O	O
and	NN	O	O
MRP	NN	O	B-protein
(	NN	O	O
MARCKS-related	NN	O	B-protein
protein	NN	O	I-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
of	NN	O	O
a	NN	O	O
third	NN	O	O
protein	NN	O	O
,	NN	O	O
with	NN	O	O
an	NN	O	O
apparent	NN	O	O
m.w.	NN	O	O
of	NN	O	O
60	NN	O	O
,	NN	O	O
000	NN	O	O
that	NN	O	O
was	NN	O	O
the	NN	O	O
most	NN	O	O
prominent	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
substrate	NN	O	I-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

p60	NN	O	B-protein
phosphorylation	NN	O	O
was	NN	O	O
time	NN	O	O
and	NN	O	O
PMA	NN	O	O
dose	NN	O	O
dependent	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
cell-permeable	NN	O	O
diacylglycerol	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
inactive	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
.	NN	O	O

Two-dimensional	NN	O	O
electrophoretic	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
phosphorylation	NN	O	O
pattern	NN	O	O
from	NN	O	O
the	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
CESS	NN	O	I-cell_line
demonstrated	NN	O	O
the	NN	O	O
identity	NN	O	O
between	NN	O	O
the	NN	O	O
p60	NN	O	B-protein
protein	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
this	NN	O	O
cell	NN	O	O
line	NN	O	O
and	NN	O	O
that	NN	O	O
expressed	NN	O	O
in	NN	O	O
B-CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

p60	NN	O	B-protein
was	NN	O	O
purified	NN	O	O
from	NN	O	O
CESS	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
peptide	NN	O	O
microsequencing	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
revealed	NN	O	O
that	NN	O	O
it	NN	O	O
was	NN	O	O
lymphocyte-specific	NN	O	B-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
LSP1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
that	NN	O	O
is	NN	O	O
here	NN	O	O
characterized	NN	O	O
as	NN	O	O
the	NN	O	O
most	NN	O	O
prominent	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
substrate	NN	O	I-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
--	NN	O	O
basic	NN	O	O
research	NN	O	O
and	NN	O	O
clinical	NN	O	O
perspectives	NN	O	O
for	NN	O	O
gastroenterology	NN	O	O
.	NN	O	O

Tissue	NN	O	O
specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
is	NN	O	O
a	NN	O	O
fascinating	NN	O	O
new	NN	O	O
field	NN	O	O
in	NN	O	O
molecular	NN	O	O
immunology	NN	O	O
.	NN	O	O

This	NN	O	O
review	NN	O	O
summarizes	NN	O	O
data	NN	O	O
on	NN	O	O
specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
promoters	NN	O	B-DNA
and	NN	O	O
enhancers	NN	O	B-DNA
by	NN	O	O
nuclear	NN	O	B-protein
trans-acting	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
structural	NN	O	O
classes	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
described	NN	O	O
and	NN	O	O
basic	NN	O	O
methods	NN	O	O
for	NN	O	O
detection	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
detailed	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
most	NN	O	O
important	NN	O	O
trans-acting	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
e.g	NN	O	O
.	NN	O	O
NF-kB	NN	O	B-protein
,	NN	O	O
NF-AT	NN	O	B-protein
and	NN	O	O
STAT	NN	O	B-protein
families	NN	O	I-protein
)	NN	O	O
and	NN	O	O
their	NN	O	O
functional	NN	O	O
importance	NN	O	O
are	NN	O	O
described	NN	O	O
.	NN	O	O

Several	NN	O	O
methods	NN	O	O
for	NN	O	O
specific	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
shown	NN	O	O
that	NN	O	O
may	NN	O	O
be	NN	O	O
relevant	NN	O	O
to	NN	O	O
treatment	NN	O	O
of	NN	O	O
human	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
are	NN	O	O
discussed	NN	O	O
with	NN	O	O
regard	NN	O	O
to	NN	O	O
their	NN	O	O
potential	NN	O	O
clinical	NN	O	O
relevance	NN	O	O
for	NN	O	O
gastroenterology	NN	O	O
.	NN	O	O

-DOCSTART-	O

Selective	NN	O	O
effects	NN	O	O
of	NN	O	O
DNA	NN	O	O
damaging	NN	O	O
agents	NN	O	O
on	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
activation	NN	O	O
and	NN	O	O
virus	NN	O	O
replication	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Much	NN	O	O
attention	NN	O	O
has	NN	O	O
recently	NN	O	O
focused	NN	O	O
on	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
UV	NN	O	O
light	NN	O	O
can	NN	O	O
activate	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
LTR	NN	O	B-DNA
activation	NN	O	O
remains	NN	O	O
obscure	NN	O	O
,	NN	O	O
several	NN	O	O
lines	NN	O	O
of	NN	O	O
investigation	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
following	NN	O	O
signaling	NN	O	O
events	NN	O	O
related	NN	O	O
to	NN	O	O
generalized	NN	O	O
DNA	NN	O	O
damage	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
present	NN	O	O
data	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
HIV	NN	O	O
LTR	NN	O	B-DNA
activation	NN	O	O
is	NN	O	O
not	NN	O	O
a	NN	O	O
general	NN	O	O
consequence	NN	O	O
of	NN	O	O
cellular	NN	O	B-DNA
DNA	NN	O	I-DNA
damage	NN	O	O
,	NN	O	O
but	NN	O	O
rather	NN	O	O
a	NN	O	O
process	NN	O	O
unique	NN	O	O
to	NN	O	O
specific	NN	O	O
genotoxic	NN	O	O
stimuli	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
it	NN	O	O
does	NN	O	O
not	NN	O	O
necessarily	NN	O	O
depend	NN	O	O
on	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
several	NN	O	O
of	NN	O	O
these	NN	O	O
agents	NN	O	O
can	NN	O	O
significantly	NN	O	O
increase	NN	O	O
HIV	NN	O	O
replication	NN	O	O
and	NN	O	O
accelerate	NN	O	O
CD4	NN	O	B-protein
-positive	NN	O	O
lymphocyte	NN	O	O
cytotoxicity	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
could	NN	O	O
have	NN	O	O
clinical	NN	O	O
significance	NN	O	O
to	NN	O	O
AIDS	NN	O	O
patients	NN	O	O
with	NN	O	O
malignancies	NN	O	O
who	NN	O	O
are	NN	O	O
undergoing	NN	O	O
radiotherapy	NN	O	O
and	NN	O	O
chemotherapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Receptors	NN	O	O
for	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-10	NN	O	I-protein
and	NN	O	I-protein
IL-6-type	NN	O	I-protein
cytokines	NN	O	I-protein
use	NN	O	O
similar	NN	O	O
signaling	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
inducing	NN	O	O
transcription	NN	O	O
through	NN	O	O
IL-6	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
for	NN	O	O
interleukin	NN	O	B-protein
10	NN	O	I-protein
(	NN	O	O
IL-10R	NN	O	B-protein
)	NN	O	O
contains	NN	O	O
two	NN	O	O
box	NN	O	B-protein
3	NN	O	I-protein
sequence	NN	O	I-protein
motifs	NN	O	I-protein
that	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
signal-transducing	NN	O	B-protein
receptor	NN	O	I-protein
subunits	NN	O	I-protein
for	NN	O	O
IL-6-type	NN	O	B-protein
cytokines	NN	O	I-protein
and	NN	O	O
noted	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
activating	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
inducing	NN	O	O
transcription	NN	O	O
through	NN	O	O
IL-6-responsive	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
IL-10R	NN	O	B-protein
has	NN	O	O
signaling	NN	O	O
functions	NN	O	O
similar	NN	O	O
to	NN	O	O
IL-6R	NN	O	B-protein
in	NN	O	O
cells	NN	O	O
normally	NN	O	O
expressing	NN	O	O
these	NN	O	O
receptors	NN	O	O
,	NN	O	O
leukocytes	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
B-	NN	O	B-cell_type
,	NN	O	I-cell_type
T-	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
NK-cell	NN	O	I-cell_type
lineages	NN	O	I-cell_type
were	NN	O	O
treated	NN	O	O
with	NN	O	O
either	NN	O	O
cytokine	NN	O	O
.	NN	O	O

Both	NN	O	O
cytokines	NN	O	B-protein
activated	NN	O	O
factors	NN	O	O
that	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
sis-inducible	NN	O	B-DNA
element	NN	O	I-DNA
and	NN	O	O
included	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
.	NN	O	O

The	NN	O	O
cell	NN	O	O
response	NN	O	O
to	NN	O	O
IL-10	NN	O	B-protein
characteristically	NN	O	O
differed	NN	O	O
from	NN	O	O
that	NN	O	O
to	NN	O	O
IL-2/IL-15	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
.	NN	O	O

The	NN	O	O
signaling	NN	O	O
capabilities	NN	O	O
of	NN	O	O
the	NN	O	O
IL-10R	NN	O	B-protein
for	NN	O	O
activating	NN	O	O
specific	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
inducing	NN	O	O
gene	NN	O	O
transcription	NN	O	O
were	NN	O	O
defined	NN	O	O
by	NN	O	O
reconstitution	NN	O	O
of	NN	O	O
receptor	NN	O	O
functions	NN	O	O
in	NN	O	O
transfected	NN	O	B-cell_line
tissue	NN	O	I-cell_line
culture	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

COS-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
co-expressing	NN	O	O
the	NN	O	O
human	NN	O	O
IL-10R	NN	O	B-protein
and	NN	O	O
individual	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
confirmed	NN	O	O
a	NN	O	O
preference	NN	O	O
of	NN	O	O
the	NN	O	O
IL-10R	NN	O	B-protein
for	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
STAT1	NN	O	B-protein
.	NN	O	O

Unlike	NN	O	O
many	NN	O	O
hematopoietin	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
the	NN	O	O
IL-10R	NN	O	B-protein
did	NN	O	O
not	NN	O	O
detectably	NN	O	O
activate	NN	O	O
STAT5	NN	O	B-protein
.	NN	O	O

The	NN	O	O
IL-10R	NN	O	B-protein
,	NN	O	O
together	NN	O	O
with	NN	O	O
reporter	NN	O	O
gene	NN	O	O
constructs	NN	O	O
containing	NN	O	O
different	NN	O	O
IL-6-responsive	NN	O	B-DNA
gene	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
reconstituted	NN	O	O
in	NN	O	O
hepatoma	NN	O	B-cell_type
cells	NN	O	I-cell_type
an	NN	O	O
induction	NN	O	O
of	NN	O	O
transcription	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
that	NN	O	O
was	NN	O	O
comparable	NN	O	O
to	NN	O	O
that	NN	O	O
by	NN	O	O
IL-6	NN	O	B-protein
.	NN	O	O

This	NN	O	O
regulation	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
appreciably	NN	O	O
modified	NN	O	O
by	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
similar	NN	O	O
actions	NN	O	O
of	NN	O	O
IL-10R	NN	O	B-protein
and	NN	O	O
IL-6R	NN	O	B-protein
on	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
endogenous	NN	O	O
IL-6-responsive	NN	O	B-DNA
genes	NN	O	I-DNA
were	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
hepatoma	NN	O	B-cell_type
cells	NN	O	I-cell_type
stably	NN	O	O
expressing	NN	O	O
the	NN	O	O
IL-10R	NN	O	B-protein
.	NN	O	O

These	NN	O	O
receptor	NN	O	O
functions	NN	O	O
required	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
box	NN	O	B-protein
3	NN	O	I-protein
motifs	NN	O	I-protein
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
the	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
IL-10R	NN	O	B-protein
constructs	NN	O	O
containing	NN	O	O
progressively	NN	O	B-protein
truncated	NN	O	I-protein
cytoplasmic	NN	O	I-protein
domains	NN	O	I-protein
.	NN	O	O

The	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
IL-10R	NN	O	B-protein
,	NN	O	O
unlike	NN	O	O
other	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
interferon	NN	O	O
receptor	NN	O	O
family	NN	O	O
,	NN	O	O
is	NN	O	O
highly	NN	O	O
effective	NN	O	O
in	NN	O	O
recruiting	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
of	NN	O	O
IL-6-type	NN	O	B-protein
cytokine	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
and	NN	O	O
modification	NN	O	O
of	NN	O	O
in	NN	O	O
vivo	NN	O	O
HLA-DR	NN	O	B-DNA
promoter	NN	O	I-DNA
occupancy	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
thymic	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
:	NN	O	O
similarities	NN	O	O
and	NN	O	O
distinctions	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
HLA-DR-constitutive	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
primary	NN	O	O
cultures	NN	O	O
of	NN	O	O
thymic	NN	O	B-cell_line
epithelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
TEC	NN	O	B-cell_line
)	NN	O	O
was	NN	O	O
analyzed	NN	O	O
.	NN	O	O

This	NN	O	O
cellular	NN	O	O
system	NN	O	O
offers	NN	O	O
the	NN	O	O
advantage	NN	O	O
that	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
induction	NN	O	O
is	NN	O	O
studied	NN	O	O
in	NN	O	O
a	NN	O	O
``	NN	O	B-cell_type
physiologic	NN	O	I-cell_type
''	NN	O	I-cell_type
cell	NN	O	I-cell_type
lineage	NN	O	I-cell_type
that	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
this	NN	O	O
expression	NN	O	O
within	NN	O	O
the	NN	O	O
thymus	NN	O	O
,	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
participate	NN	O	O
to	NN	O	O
the	NN	O	O
selection	NN	O	O
and	NN	O	O
maturation	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
de	NN	O	O
novo	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
CIITA	NN	O	B-protein
trans-activator	NN	O	I-protein
,	NN	O	O
a	NN	O	O
crucial	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
gene	NN	O	I-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
anatomy	NN	O	O
of	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
DRA	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
corresponding	NN	O	O
binding	NN	O	B-protein
factors	NN	O	I-protein
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
in	NN	O	O
vivo	NN	O	O
DNAse	NN	O	B-protein
I	NN	O	I-protein
footprint	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
treatment	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
induces	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
occupancy	NN	O	O
of	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
gene	NN	O	I-DNA
regulatory	NN	O	I-DNA
sequences	NN	O	I-DNA
by	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
resulting	NN	O	O
occupancy	NN	O	O
displays	NN	O	O
strong	NN	O	O
similarities	NN	O	O
with	NN	O	O
the	NN	O	O
one	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
MHC	NN	O	B-cell_type
class	NN	O	I-cell_type
II-constitutive	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
represented	NN	O	O
by	NN	O	O
both	NN	O	O
the	NN	O	O
Burkitt	NN	O	B-cell_line
lymphoma	NN	O	I-cell_line
line	NN	O	I-cell_line
Raji	NN	O	I-cell_line
and	NN	O	O
normal	NN	O	B-cell_type
tonsil-	NN	O	I-cell_type
derived	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
some	NN	O	O
peculiar	NN	O	O
differences	NN	O	O
were	NN	O	O
observed	NN	O	O
between	NN	O	O
the	NN	O	O
TEC	NN	O	B-cell_line
,	NN	O	O
either	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
or	NN	O	O
not	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
constitutive	NN	O	O
B	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
both	NN	O	O
common	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
one	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
CIITA	NN	O	B-protein
trans-activator	NN	O	I-protein
,	NN	O	O
and	NN	O	O
distinct	NN	O	O
tissue-specific	NN	O	O
constraints	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
constitutive	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
E3330	NN	O	O
,	NN	O	O
a	NN	O	O
novel	NN	O	O
quinone	NN	O	O
derivative	NN	O	O
able	NN	O	O
to	NN	O	O
suppress	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
generation	NN	O	O
,	NN	O	O
on	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

(	NN	O	O
2E	NN	O	O
)	NN	O	O
-3-	NN	O	O
[	NN	O	O
5-	NN	O	O
(	NN	O	O
2	NN	O	O
,	NN	O	O
3-Dimethoxy-6-methyl-1	NN	O	O
,	NN	O	O
4-benzoquinoyl	NN	O	O
)	NN	O	O
]	NN	O	O
-2-nonyl-2-	NN	O	O
propenoic	NN	O	O
acid	NN	O	O
(	NN	O	O
E3330	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
novel	NN	O	O
agent	NN	O	O
with	NN	O	O
hepatoprotective	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
E3330	NN	O	O
on	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
TNF	NN	O	I-DNA
)	NN	O	I-DNA
-alpha	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
on	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
run-on	NN	O	O
experiments	NN	O	O
showed	NN	O	O
that	NN	O	O
E3330	NN	O	O
decreases	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-DNA
gene	NN	O	I-DNA
induced	NN	O	O
by	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
stimulation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
we	NN	O	O
constructed	NN	O	O
a	NN	O	O
secreted-type	NN	O	B-DNA
placental	NN	O	I-DNA
alkaline	NN	O	I-DNA
phosphatase	NN	O	I-DNA
(	NN	O	I-DNA
PLAP	NN	O	I-DNA
)	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
whose	NN	O	O
transcription	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
1.4-kb	NN	O	B-DNA
human	NN	O	I-DNA
TNF-alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
stable	NN	O	O
transformant	NN	O	O
of	NN	O	O
the	NN	O	O
PLAP	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
derived	NN	O	O
from	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
showed	NN	O	O
very	NN	O	O
little	NN	O	O
activity	NN	O	O
on	NN	O	O
the	NN	O	O
promoter	NN	O	O
before	NN	O	O
stimulation	NN	O	O
,	NN	O	O
whereas	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
dramatic	NN	O	O
increase	NN	O	O
in	NN	O	O
PLAP	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

E3330	NN	O	O
inhibited	NN	O	O
this	NN	O	O
induced	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

There	NN	O	O
are	NN	O	O
four	NN	O	O
putative	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
(	NN	O	O
kappa	NN	O	B-DNA
B-1	NN	O	I-DNA
,	NN	O	O
kappa	NN	O	B-DNA
B-2	NN	O	I-DNA
,	NN	O	O
kappa	NN	O	B-DNA
B-3	NN	O	I-DNA
,	NN	O	O
kappa	NN	O	B-DNA
B-4	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
human	NN	O	B-DNA
TNF-alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

By	NN	O	O
using	NN	O	O
mutated	NN	O	B-DNA
promoter-PLAP	NN	O	I-DNA
plasmids	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
established	NN	O	O
that	NN	O	O
these	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
were	NN	O	O
necessary	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
transcription	NN	O	O
on	NN	O	O
stimulation	NN	O	O
with	NN	O	O
LPS	NN	O	O
.	NN	O	O

A	NN	O	O
gel	NN	O	O
retardation	NN	O	O
experiment	NN	O	O
with	NN	O	O
synthetic	NN	O	O
double-stranded	NN	O	O
oligonucleotides	NN	O	O
showed	NN	O	O
that	NN	O	O
activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
consisting	NN	O	O
of	NN	O	O
p50/p65	NN	O	B-protein
heterodimer	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
all	NN	O	O
four	NN	O	O
putative	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
DNA	NN	O	I-DNA
probes	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
all	NN	O	O
four	NN	O	O
putative	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
recognition	NN	O	I-DNA
sites	NN	O	I-DNA
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
inducible	NN	O	O
TNF-alpha	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

E3330	NN	O	O
decreased	NN	O	O
activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
nuclei	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
E3330	NN	O	O
inhibits	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and/or	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclei	NN	O	O
.	NN	O	O

Western	NN	O	O
blotting	NN	O	O
analysis	NN	O	O
with	NN	O	O
anti-I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
antibody	NN	O	I-protein
indicated	NN	O	O
that	NN	O	O
E3330	NN	O	O
inhibited	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
an	NN	O	O
inhibitory	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
in	NN	O	O
LPS-stimulated	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
.	NN	O	O

E3330	NN	O	O
may	NN	O	O
suppress	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
active	NN	O	O
oxygen	NN	O	O
species	NN	O	O
serving	NN	O	O
as	NN	O	O
common	NN	O	O
messengers	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

C-terminal	NN	O	B-protein
activating	NN	O	I-protein
and	NN	O	I-protein
inhibitory	NN	O	I-protein
domains	NN	O	I-protein
determine	NN	O	O
the	NN	O	O
transactivation	NN	O	O
potential	NN	O	O
of	NN	O	O
BSAP	NN	O	B-protein
(	NN	O	O
Pax-5	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
Pax-2	NN	O	B-DNA
and	NN	O	O
Pax-8	NN	O	B-DNA
.	NN	O	O

Pax-5	NN	O	B-DNA
encodes	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
BSAP	NN	O	B-protein
which	NN	O	O
plays	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
in	NN	O	O
early	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
and	NN	O	O
midbrain	NN	O	O
patterning	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
have	NN	O	O
analysed	NN	O	O
the	NN	O	O
structural	NN	O	O
requirements	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
BSAP	NN	O	B-protein
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
mutagenesis	NN	O	O
and	NN	O	O
transient	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
serine/threonine/proline-rich	NN	O	I-protein
region	NN	O	I-protein
of	NN	O	O
BSAP	NN	O	B-protein
contains	NN	O	O
a	NN	O	O
potent	NN	O	O
transactivation	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
55	NN	O	O
amino	NN	O	O
acids	NN	O	O
which	NN	O	O
is	NN	O	O
active	NN	O	O
from	NN	O	O
promoter	NN	O	B-DNA
and	NN	O	I-DNA
enhancer	NN	O	I-DNA
positions	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
transactivation	NN	O	B-protein
domain	NN	O	I-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
inactivated	NN	O	O
by	NN	O	O
a	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
frameshift	NN	O	O
mutation	NN	O	O
in	NN	O	O
one	NN	O	O
PAX-5	NN	O	B-DNA
allele	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
acute	NN	O	B-cell_line
lymphoblastic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
REH	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
transactivation	NN	O	B-protein
domain	NN	O	I-protein
is	NN	O	O
negatively	NN	O	O
regulated	NN	O	O
by	NN	O	O
adjacent	NN	O	O
sequences	NN	O	O
from	NN	O	O
the	NN	O	O
extreme	NN	O	B-protein
C-terminus	NN	O	I-protein
.	NN	O	O

The	NN	O	O
activating	NN	O	O
and	NN	O	O
inhibitory	NN	O	O
domains	NN	O	O
function	NN	O	O
together	NN	O	O
as	NN	O	O
an	NN	O	O
independent	NN	O	O
regulatory	NN	O	O
module	NN	O	O
in	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
fusion	NN	O	O
to	NN	O	O
the	NN	O	O
GAL4	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

The	NN	O	O
same	NN	O	O
arrangement	NN	O	O
of	NN	O	O
positively	NN	O	B-DNA
and	NN	O	I-DNA
negatively	NN	O	I-DNA
acting	NN	O	I-DNA
sequences	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
conserved	NN	O	O
in	NN	O	O
the	NN	O	O
mammalian	NN	O	B-DNA
Pax-2	NN	O	I-DNA
and	NN	O	O
Pax-8	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
zebrafish	NN	O	B-DNA
Pax-b	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
sea	NN	O	B-protein
urchin	NN	O	I-protein
Pax-258	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
competence	NN	O	O
of	NN	O	O
a	NN	O	O
subfamily	NN	O	O
of	NN	O	O
Pax	NN	O	B-protein
proteins	NN	O	I-protein
is	NN	O	O
determined	NN	O	O
by	NN	O	O
a	NN	O	O
C-terminal	NN	O	B-protein
regulatory	NN	O	I-protein
module	NN	O	I-protein
composed	NN	O	O
of	NN	O	O
activating	NN	O	B-DNA
and	NN	O	I-DNA
inhibitory	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Second	NN	O	O
messenger	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
is	NN	O	O
absent	NN	O	O
in	NN	O	O
androgen	NN	O	B-cell_line
insensitive	NN	O	I-cell_line
human	NN	O	I-cell_line
prostatic	NN	O	I-cell_line
carcinoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
PC-3	NN	O	B-cell_line
and	NN	O	O
DU-145	NN	O	B-cell_line
.	NN	O	O

A	NN	O	O
theoretical	NN	O	O
pathway	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
AR	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
via	NN	O	O
a	NN	O	O
cAMP	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CRE	NN	O	B-DNA
)	NN	O	O
present	NN	O	O
in	NN	O	O
its	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
(	NN	O	O
-508	NN	O	O
to	NN	O	O
-501	NN	O	O
)	NN	O	O
.	NN	O	O

After	NN	O	O
20	NN	O	O
h	NN	O	O
of	NN	O	O
stimulation	NN	O	O
with	NN	O	O
8-bromo-cAMP	NN	O	O
,	NN	O	O
AR	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
upregulated	NN	O	O
in	NN	O	O
LNCaP	NN	O	B-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
either	NN	O	O
PC-3	NN	O	B-cell_line
or	NN	O	O
DU-145	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
CRE	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
was	NN	O	O
the	NN	O	O
same	NN	O	O
in	NN	O	O
all	NN	O	O
cell	NN	O	O
lines	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
putative	NN	O	O
AR-CRE	NN	O	B-protein
forms	NN	O	O
specific	NN	O	O
and	NN	O	O
compatible	NN	O	O
protein	NN	O	O
interactions	NN	O	O
with	NN	O	O
CREB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
ability	NN	O	O
to	NN	O	O
regulate	NN	O	O
AR	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
via	NN	O	O
the	NN	O	O
second	NN	O	O
messenger	NN	O	O
pathway	NN	O	O
is	NN	O	O
lost	NN	O	O
in	NN	O	O
the	NN	O	O
PC-3	NN	O	B-cell_line
and	NN	O	I-cell_line
DU-145	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
primary	NN	O	O
mechanism	NN	O	O
of	NN	O	O
androgen	NN	O	O
insensitivity	NN	O	O
in	NN	O	O
prostate	NN	O	O
cancer	NN	O	O
.	NN	O	O

-DOCSTART-	O

Structural	NN	O	O
and	NN	O	O
functional	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD36	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
:	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
proximal	NN	O	O
PEBP2/CBF	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

CD36	NN	O	B-protein
is	NN	O	O
a	NN	O	O
cell	NN	O	O
surface	NN	O	O
glycoprotein	NN	O	B-protein
composed	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
polypeptide	NN	O	O
chain	NN	O	O
,	NN	O	O
which	NN	O	O
interacts	NN	O	O
with	NN	O	O
thrombospondin	NN	O	B-protein
,	NN	O	O
collagens	NN	O	B-protein
type	NN	O	I-protein
I	NN	O	I-protein
and	NN	O	I-protein
IV	NN	O	I-protein
,	NN	O	O
oxidized	NN	O	O
low	NN	O	B-protein
density	NN	O	I-protein
lipoprotein	NN	O	I-protein
,	NN	O	O
fatty	NN	O	O
acids	NN	O	O
,	NN	O	O
anionic	NN	O	O
phospholipids	NN	O	O
,	NN	O	O
and	NN	O	O
erythrocytes	NN	O	B-cell_type
parasitized	NN	O	O
with	NN	O	O
Plasmodium	NN	O	O
falciparum	NN	O	O
.	NN	O	O

Its	NN	O	O
expression	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
a	NN	O	O
few	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
including	NN	O	O
monocyte/macrophages	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
CD36	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
phagocytosis	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
foam	NN	O	O
cell	NN	O	O
formation	NN	O	O
by	NN	O	O
uptake	NN	O	O
of	NN	O	O
oxidized	NN	O	B-protein
low	NN	O	I-protein
density	NN	O	I-protein
lipoprotein	NN	O	I-protein
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
CD36	NN	O	B-protein
gene	NN	O	O
in	NN	O	O
monocytic	NN	O	O
cells	NN	O	O
we	NN	O	O
have	NN	O	O
isolated	NN	O	O
and	NN	O	O
analyzed	NN	O	O
the	NN	O	O
CD36	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
experiments	NN	O	O
of	NN	O	O
5'-deletion	NN	O	B-DNA
fragments	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CD36	NN	O	B-DNA
promoter	NN	O	I-DNA
coupled	NN	O	O
to	NN	O	O
luciferase	NN	O	B-protein
demonstrated	NN	O	O
that	NN	O	O
as	NN	O	O
few	NN	O	O
as	NN	O	O
158	NN	O	B-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
upstream	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
were	NN	O	O
sufficient	NN	O	O
to	NN	O	O
direct	NN	O	O
the	NN	O	O
monocyte-specific	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Within	NN	O	O
the	NN	O	O
above	NN	O	O
region	NN	O	O
,	NN	O	O
the	NN	O	O
fragment	NN	O	O
spanning	NN	O	O
nucleotides	NN	O	B-DNA
-158	NN	O	I-DNA
to	NN	O	I-DNA
-90	NN	O	I-DNA
was	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
transcription	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Biochemical	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
region	NN	O	O
-158/-90	NN	O	B-DNA
revealed	NN	O	O
a	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
the	NN	O	O
polyomavirus	NN	O	B-protein
enhancer-binding	NN	O	I-protein
protein	NN	O	I-protein
2/core-binding	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
PEBP2/CBF	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
at	NN	O	O
position	NN	O	B-DNA
-103	NN	O	I-DNA
.	NN	O	O

Disruption	NN	O	O
of	NN	O	O
the	NN	O	O
PEBP2/CBF	NN	O	B-DNA
site	NN	O	I-DNA
markedly	NN	O	O
diminished	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
PEBP2/CBF	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
constitutive	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
CD36	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
PEBP2/CBF	NN	O	B-protein
family	NN	O	I-protein
in	NN	O	O
chromosome	NN	O	O
translocations	NN	O	O
associated	NN	O	O
with	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
myeloid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
encoding	NN	O	O
for	NN	O	O
myeloperoxidase	NN	O	B-protein
,	NN	O	O
elastase	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
colony-stimulating	NN	O	B-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
highlights	NN	O	O
the	NN	O	O
relevance	NN	O	O
of	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
CD36	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
PEBP2/CBF	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

HLA-DQB1	NN	O	B-DNA
codon	NN	O	I-DNA
57	NN	O	I-DNA
is	NN	O	O
critical	NN	O	O
for	NN	O	O
peptide	NN	O	O
binding	NN	O	O
and	NN	O	O
recognition	NN	O	O
.	NN	O	O

The	NN	O	O
association	NN	O	O
of	NN	O	O
specific	NN	O	O
HLA-DQ	NN	O	B-DNA
alleles	NN	O	I-DNA
with	NN	O	O
autoimmunity	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
discrete	NN	O	O
polymorphisms	NN	O	O
in	NN	O	O
the	NN	O	O
HLA-DQ	NN	O	B-DNA
sequence	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
localized	NN	O	O
within	NN	O	O
sites	NN	O	O
suitable	NN	O	O
for	NN	O	O
peptide	NN	O	O
recognition	NN	O	O
.	NN	O	O

The	NN	O	O
polymorphism	NN	O	O
at	NN	O	O
residue	NN	O	O
57	NN	O	O
of	NN	O	O
the	NN	O	O
DQB1	NN	O	O
polypeptide	NN	O	O
is	NN	O	O
of	NN	O	O
particular	NN	O	O
interest	NN	O	O
since	NN	O	O
it	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
major	NN	O	O
structural	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
salt	NN	O	B-protein
bridge	NN	O	I-protein
structure	NN	O	I-protein
at	NN	O	O
one	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
peptide-binding	NN	O	B-protein
cleft	NN	O	I-protein
of	NN	O	O
the	NN	O	O
DQ	NN	O	O
molecules	NN	O	O
.	NN	O	O

This	NN	O	O
polymorphism	NN	O	O
at	NN	O	O
residue	NN	O	O
57	NN	O	O
is	NN	O	O
a	NN	O	O
recurrent	NN	O	O
feature	NN	O	O
of	NN	O	O
HLA-DQ	NN	O	O
evolution	NN	O	O
,	NN	O	O
occurring	NN	O	O
in	NN	O	O
multiple	NN	O	O
distinct	NN	O	O
allelic	NN	O	O
families	NN	O	O
,	NN	O	O
which	NN	O	O
implies	NN	O	O
a	NN	O	O
functional	NN	O	O
selection	NN	O	O
for	NN	O	O
maintaining	NN	O	O
variation	NN	O	O
at	NN	O	O
this	NN	O	O
position	NN	O	O
in	NN	O	O
the	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecule	NN	O	I-protein
.	NN	O	O

We	NN	O	O
directly	NN	O	O
tested	NN	O	O
the	NN	O	O
amino	NN	O	O
acid	NN	O	O
polymorphism	NN	O	O
at	NN	O	O
this	NN	O	O
site	NN	O	O
as	NN	O	O
a	NN	O	O
determinant	NN	O	O
for	NN	O	O
peptide	NN	O	O
binding	NN	O	O
and	NN	O	O
for	NN	O	O
antigen-specific	NN	O	O
T	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
a	NN	O	O
single	NN	O	O
Ala	NN	O	O
--	NN	O	O
>	NN	O	O
Asp	NN	O	O
amino	NN	O	O
acid	NN	O	O
57	NN	O	O
substitution	NN	O	O
in	NN	O	O
an	NN	O	O
HLA-DQ3.2	NN	O	O
molecule	NN	O	O
regulated	NN	O	O
binding	NN	O	O
of	NN	O	O
an	NN	O	O
HSV-2	NN	O	O
VP-16-derived	NN	O	O
peptide	NN	O	O
.	NN	O	O

A	NN	O	O
complementary	NN	O	O
single-residue	NN	O	O
substitution	NN	O	O
in	NN	O	O
the	NN	O	O
peptide	NN	O	O
abolished	NN	O	O
its	NN	O	O
binding	NN	O	O
to	NN	O	O
DQ3.2	NN	O	O
and	NN	O	O
converted	NN	O	O
it	NN	O	O
to	NN	O	O
a	NN	O	O
peptide	NN	O	O
that	NN	O	O
can	NN	O	O
bind	NN	O	O
to	NN	O	O
DQ3.1	NN	O	O
and	NN	O	O
DQ3.3	NN	O	B-protein
Asp-57-positive	NN	O	I-protein
MHC	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

These	NN	O	O
binding	NN	O	O
studies	NN	O	O
were	NN	O	O
paralleled	NN	O	O
by	NN	O	O
specific	NN	O	O
T	NN	O	O
cell	NN	O	O
recognition	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-protein
II-peptide	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
substituted	NN	O	O
peptide	NN	O	O
abolished	NN	O	O
T	NN	O	O
cell	NN	O	O
reactivity	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
directed	NN	O	O
to	NN	O	O
the	NN	O	O
DQ3.2-peptide	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
same	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
recognized	NN	O	O
the	NN	O	O
substituted	NN	O	O
peptide	NN	O	O
presented	NN	O	O
by	NN	O	O
DQ3.3	NN	O	O
,	NN	O	O
a	NN	O	O
class	NN	O	O
II	NN	O	O
restriction	NN	O	O
element	NN	O	O
differing	NN	O	O
from	NN	O	O
DQ3.2	NN	O	O
only	NN	O	O
at	NN	O	O
residue	NN	O	O
57	NN	O	O
.	NN	O	O

This	NN	O	O
structural	NN	O	O
and	NN	O	O
functional	NN	O	O
complementarity	NN	O	O
for	NN	O	O
residue	NN	O	O
57	NN	O	O
and	NN	O	O
a	NN	O	O
specific	NN	O	O
peptide	NN	O	O
residue	NN	O	O
identifies	NN	O	O
this	NN	O	O
interaction	NN	O	O
as	NN	O	O
a	NN	O	O
key	NN	O	O
controlling	NN	O	O
determinant	NN	O	O
of	NN	O	O
restricted	NN	O	O
recognition	NN	O	O
in	NN	O	O
HLA-DQ-specific	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Autocrine	NN	O	O
activation	NN	O	O
by	NN	O	O
interferon-gamma	NN	O	B-protein
of	NN	O	O
STAT	NN	O	B-protein
factors	NN	O	I-protein
following	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
engagement	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor/CD3	NN	O	I-protein
complex	NN	O	I-protein
and	NN	O	O
co-stimulatory	NN	O	B-protein
molecules	NN	O	I-protein
,	NN	O	O
and	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
triggering	NN	O	O
of	NN	O	O
several	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
which	NN	O	O
lead	NN	O	O
rapidly	NN	O	O
to	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

A	NN	O	O
result	NN	O	O
of	NN	O	O
this	NN	O	O
activation	NN	O	O
process	NN	O	O
is	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
genes	NN	O	O
,	NN	O	O
including	NN	O	O
those	NN	O	O
encoding	NN	O	O
cytokines	NN	O	B-protein
such	NN	O	O
as	NN	O	O
interleukin-2	NN	O	B-protein
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
,	NN	O	O
and	NN	O	O
interferon	NN	O	B-protein
(	NN	O	I-protein
IFN	NN	O	I-protein
)	NN	O	I-protein
-gamma	NN	O	I-protein
which	NN	O	O
have	NN	O	O
important	NN	O	O
immunoregulatory	NN	O	B-protein
effects	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
factor	NN	O	I-protein
containing	NN	O	O
STAT1	NN	O	B-protein
also	NN	O	O
becomes	NN	O	O
activated	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
or	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
not	NN	O	O
until	NN	O	O
1-2	NN	O	O
h	NN	O	O
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Activation	NN	O	O
is	NN	O	O
delayed	NN	O	O
a	NN	O	O
further	NN	O	O
1-2	NN	O	O
hr	NN	O	O
when	NN	O	O
mononuclear	NN	O	B-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
are	NN	O	O
stimulated	NN	O	O
by	NN	O	O
an	NN	O	O
antigen	NN	O	O
which	NN	O	O
requires	NN	O	O
processing	NN	O	O
.	NN	O	O

Appearance	NN	O	O
of	NN	O	O
the	NN	O	O
STAT1	NN	O	B-protein
factor	NN	O	O
is	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
and	NN	O	O
blocked	NN	O	O
by	NN	O	O
cycloheximide	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
its	NN	O	O
activation	NN	O	O
is	NN	O	O
dependent	NN	O	O
upon	NN	O	O
a	NN	O	O
protein	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
synthesized	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
initial	NN	O	O
signaling	NN	O	O
events	NN	O	O
.	NN	O	O

Neutralizing	NN	O	O
antiserum	NN	O	O
against	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
tested	NN	O	O
,	NN	O	O
blocked	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
factor	NN	O	O
almost	NN	O	O
completely	NN	O	O
,	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
was	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
culture	NN	O	O
supernatants	NN	O	O
of	NN	O	O
stimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
at	NN	O	O
levels	NN	O	O
at	NN	O	O
which	NN	O	O
recombinant	NN	O	O
IFN-gamma	NN	O	B-protein
could	NN	O	O
activate	NN	O	O
the	NN	O	O
factor	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
a	NN	O	O
STAT1	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
activated	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
synthesized	NN	O	O
and	NN	O	O
released	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
populations	NN	O	I-cell_type
.	NN	O	O

While	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
both	NN	O	O
secrete	NN	O	O
and	NN	O	O
respond	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
an	NN	O	O
autocrine	NN	O	O
loop	NN	O	O
,	NN	O	O
it	NN	O	O
seems	NN	O	O
likely	NN	O	O
that	NN	O	O
the	NN	O	O
responding	NN	O	O
cells	NN	O	O
may	NN	O	O
differ	NN	O	O
from	NN	O	O
those	NN	O	O
synthesizing	NN	O	O
this	NN	O	O
cytokine	NN	O	B-protein
in	NN	O	O
the	NN	O	O
mononuclear	NN	O	B-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
in	NN	O	O
the	NN	O	O
light	NN	O	O
of	NN	O	O
the	NN	O	O
recent	NN	O	O
report	NN	O	O
that	NN	O	O
Th1	NN	O	B-cell_line
cells	NN	O	I-cell_line
lack	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
receptor	NN	O	O
chain	NN	O	O
necessary	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
(	NN	O	O
Pernis	NN	O	O
,	NN	O	O
A.	NN	O	O
,	NN	O	O
Gupta	NN	O	O
,	NN	O	O
S.	NN	O	O
,	NN	O	O
Gollob	NN	O	O
,	NN	O	O
K.J.	NN	O	O
,	NN	O	O
Garfein	NN	O	O
,	NN	O	O
E.	NN	O	O
,	NN	O	O
Coffman	NN	O	O
,	NN	O	O
R.L.	NN	O	O
,	NN	O	O
Schindler	NN	O	O
,	NN	O	O
C.	NN	O	O
,	NN	O	O
and	NN	O	O
Rothman	NN	O	O
,	NN	O	O
P.	NN	O	O
,	NN	O	O
Science	NN	O	O
1995.	NN	O	O
269	NN	O	O
:	NN	O	O
245	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Absence	NN	O	O
of	NN	O	O
T-cell-	NN	O	B-protein
and	NN	O	I-protein
B-cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
TCF-1	NN	O	B-protein
,	NN	O	O
GATA-3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
BSAP	NN	O	B-protein
in	NN	O	O
Hodgkin	NN	O	B-cell_line
's	NN	O	I-cell_line
Reed-Sternberg	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Based	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
T	NN	O	B-DNA
cell	NN	O	I-DNA
receptor-beta	NN	O	I-DNA
(	NN	O	I-DNA
TcR-beta	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
rearrangements	NN	O	I-DNA
in	NN	O	O
L428	NN	O	B-cell_line
and	NN	O	I-cell_line
HDLM-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
in	NN	O	O
HDLM-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
(	NN	O	I-DNA
IgH	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
rearrangement	NN	O	I-DNA
in	NN	O	O
KM-H2	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
some	NN	O	O
researchers	NN	O	O
have	NN	O	O
concluded	NN	O	O
that	NN	O	O
these	NN	O	O
long-term	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
are	NN	O	O
lymphoid	NN	O	O
in	NN	O	O
nature	NN	O	O
.	NN	O	O

The	NN	O	O
information	NN	O	O
obtained	NN	O	O
from	NN	O	O
these	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
has	NN	O	O
also	NN	O	O
been	NN	O	O
used	NN	O	O
in	NN	O	O
arguments	NN	O	O
for	NN	O	O
a	NN	O	O
lymphoid	NN	O	O
origin	NN	O	O
of	NN	O	O
H-RS	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
tissue	NN	O	O
despite	NN	O	O
the	NN	O	O
frequent	NN	O	O
absence	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-protein
markers	NN	O	I-protein
and	NN	O	O
Ig/TcR	NN	O	B-DNA
gene	NN	O	I-DNA
rearrangements	NN	O	I-DNA
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

We	NN	O	O
questioned	NN	O	O
whether	NN	O	O
one	NN	O	O
can	NN	O	O
use	NN	O	O
the	NN	O	O
limited	NN	O	O
expression	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-protein
markers	NN	O	I-protein
or	NN	O	O
the	NN	O	O
limited	NN	O	O
gene	NN	O	O
rearrangement	NN	O	O
to	NN	O	O
conclude	NN	O	O
that	NN	O	O
H-RS	NN	O	B-cell_line
cells	NN	O	I-cell_line
have	NN	O	O
a	NN	O	O
lymphoid	NN	O	O
origin	NN	O	O
,	NN	O	O
because	NN	O	O
these	NN	O	O
markers	NN	O	O
may	NN	O	O
be	NN	O	O
aberrant	NN	O	O
in	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
two	NN	O	O
T-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
(	NN	O	O
TCF-1	NN	O	B-protein
and	NN	O	O
GATA-3	NN	O	B-protein
)	NN	O	O
and	NN	O	O
one	NN	O	O
B-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
BSAP	NN	O	B-protein
)	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_line
H-RS	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
using	NN	O	O
a	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

The	NN	O	O
sensitivity	NN	O	O
and	NN	O	O
specificity	NN	O	O
of	NN	O	O
this	NN	O	O
assay	NN	O	O
for	NN	O	O
determination	NN	O	O
of	NN	O	O
cell	NN	O	B-cell_line
lineage	NN	O	I-cell_line
have	NN	O	O
been	NN	O	O
established	NN	O	O
in	NN	O	O
a	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
and	NN	O	I-cell_line
murine	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

All	NN	O	O
three	NN	O	O
types	NN	O	O
of	NN	O	O
H-RS	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
consistently	NN	O	O
negative	NN	O	O
for	NN	O	O
BSAP	NN	O	B-protein
,	NN	O	O
TCF-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GATA-3	NN	O	B-protein
.	NN	O	O

The	NN	O	O
absence	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
was	NN	O	O
confirmed	NN	O	O
in	NN	O	O
H-RS	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
tissues	NN	O	O
by	NN	O	O
an	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
technique	NN	O	O
.	NN	O	O

Virtually	NN	O	O
all	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
some	NN	O	O
myeloma	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
are	NN	O	O
positive	NN	O	O
for	NN	O	O
BSAP	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
for	NN	O	O
promoters	NN	O	B-DNA
for	NN	O	O
several	NN	O	O
B-cell	NN	O	B-protein
markers	NN	O	I-protein
,	NN	O	O
including	NN	O	O
VpreB1	NN	O	B-protein
,	NN	O	O
lambda	NN	O	B-protein
5	NN	O	I-protein
,	NN	O	O
CD19	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CD20	NN	O	B-protein
.	NN	O	O

All	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
tested	NN	O	O
were	NN	O	O
positive	NN	O	O
for	NN	O	O
TCF-1	NN	O	B-protein
and	NN	O	O
GATA-3	NN	O	B-protein
,	NN	O	O
which	NN	O	O
are	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
for	NN	O	O
promoters	NN	O	O
for	NN	O	O
several	NN	O	O
T-cell-restricted	NN	O	B-protein
markers	NN	O	I-protein
,	NN	O	O
including	NN	O	O
CD2	NN	O	B-protein
,	NN	O	O
CD3	NN	O	B-protein
,	NN	O	O
TcR	NN	O	B-protein
,	NN	O	O
and	NN	O	O
lck	NN	O	B-protein
.	NN	O	O

The	NN	O	O
absence	NN	O	O
of	NN	O	O
BSAP	NN	O	B-protein
,	NN	O	O
TCF-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GATA-3	NN	O	B-protein
clearly	NN	O	O
indicates	NN	O	O
an	NN	O	O
underlying	NN	O	O
difference	NN	O	O
between	NN	O	O
H-RS	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

MEK1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinases	NN	O	I-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

TCR	NN	O	B-protein
engagement	NN	O	O
stimulates	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
Raf-1	NN	O	B-protein
.	NN	O	O

Active	NN	O	O
Raf-1	NN	O	B-protein
phosphorylates	NN	O	O
and	NN	O	O
activates	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MAP	NN	O	I-protein
)	NN	O	I-protein
kinase/extracellular	NN	O	I-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
kinase	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
MEK1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
phosphorylates	NN	O	O
and	NN	O	O
activates	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinases/extracellular	NN	O	I-protein
signal	NN	O	I-protein
regulated	NN	O	I-protein
kinases	NN	O	I-protein
,	NN	O	O
ERK1	NN	O	B-protein
and	NN	O	O
ERK2	NN	O	B-protein
.	NN	O	O

Raf-1	NN	O	B-protein
activity	NN	O	O
promotes	NN	O	O
IL-2	NN	O	O
production	NN	O	O
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
sought	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
MEK1	NN	O	B-protein
and	NN	O	O
ERK	NN	O	B-protein
activities	NN	O	O
also	NN	O	O
stimulate	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
constitutively	NN	O	O
active	NN	O	O
Raf-1	NN	O	B-protein
or	NN	O	O
MEK1	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
enhanced	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
-driven	NN	O	O
transcription	NN	O	O
stimulated	NN	O	O
by	NN	O	O
a	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
and	NN	O	O
PMA	NN	O	O
,	NN	O	O
and	NN	O	O
together	NN	O	O
with	NN	O	O
a	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
each	NN	O	O
protein	NN	O	O
was	NN	O	O
sufficient	NN	O	O
to	NN	O	O
stimulate	NN	O	O
NF-AT	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
MEK1-interfering	NN	O	B-protein
mutants	NN	O	I-protein
inhibited	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
-driven	NN	O	O
transcription	NN	O	O
and	NN	O	O
blocked	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
constitutively	NN	O	O
active	NN	O	O
Ras	NN	O	B-protein
and	NN	O	O
Raf-1	NN	O	B-protein
to	NN	O	O
costimulate	NN	O	O
NF-AT	NN	O	B-protein
activity	NN	O	O
with	NN	O	O
a	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinase-specific	NN	O	I-protein
phosphatase	NN	O	I-protein
,	NN	O	O
MKP-1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
blocks	NN	O	O
ERK	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
inhibited	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
NF-AT	NN	O	B-protein
-driven	NN	O	O
transcription	NN	O	O
stimulated	NN	O	O
by	NN	O	O
a	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
and	NN	O	O
PMA	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
addition	NN	O	O
,	NN	O	O
MKP-1	NN	O	B-protein
neutralized	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
enhancement	NN	O	O
caused	NN	O	O
by	NN	O	O
active	NN	O	O
Raf-1	NN	O	O
and	NN	O	O
MEK1	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
consisting	NN	O	O
of	NN	O	O
Raf-1	NN	O	B-protein
,	NN	O	O
MEK1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
ERK1	NN	O	B-protein
and	NN	O	O
ERK2	NN	O	B-protein
functions	NN	O	O
in	NN	O	O
the	NN	O	O
stimulation	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Multiple	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
interleukin	NN	O	I-DNA
9	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
genetic	NN	O	O
elements	NN	O	O
and	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
responsible	NN	O	O
for	NN	O	O
human	NN	O	B-DNA
interleukin	NN	O	I-DNA
9	NN	O	I-DNA
(	NN	O	I-DNA
IL-9	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
virus	NN	O	I-cell_line
type	NN	O	I-cell_line
I-transformed	NN	O	I-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
C5MJ2	NN	O	B-cell_line
,	NN	O	O
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
IL-9	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
is	NN	O	O
controlled	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
serially	NN	O	O
deleted	NN	O	O
sequences	NN	O	O
of	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-9	NN	O	B-DNA
gene	NN	O	I-DNA
has	NN	O	O
revealed	NN	O	O
several	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
the	NN	O	O
basal	NN	O	O
and	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-9	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
C5MJ2	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

An	NN	O	O
AP-1	NN	O	O
site	NN	O	O
at	NN	O	O
-146	NN	O	O
to	NN	O	O
-140	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-9	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
proximal	NN	O	B-DNA
region	NN	O	I-DNA
between	NN	O	O
-46	NN	O	B-DNA
and	NN	O	I-DNA
-80	NN	O	I-DNA
was	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
minimum	NN	O	O
sequence	NN	O	O
for	NN	O	O
the	NN	O	O
basal	NN	O	O
and	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-9	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
C5MJ2	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Within	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
an	NN	O	O
NF-kappaB	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
-59	NN	O	B-DNA
to	NN	O	I-DNA
-50	NN	O	I-DNA
and	NN	O	O
its	NN	O	O
adjacent	NN	O	O
20-base	NN	O	B-DNA
pair	NN	O	I-DNA
upstream	NN	O	I-DNA
sequence	NN	O	I-DNA
were	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
IL-9	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

DNA-protein	NN	O	O
binding	NN	O	O
studies	NN	O	O
indicated	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
and	NN	O	O
potentially	NN	O	O
novel	NN	O	B-protein
proteins	NN	O	I-protein
(	NN	O	O
around	NN	O	O
35	NN	O	B-protein
kDa	NN	O	I-protein
)	NN	O	O
can	NN	O	O
bind	NN	O	O
to	NN	O	O
this	NN	O	O
important	NN	O	O
sequence	NN	O	O
.	NN	O	O

Mutations	NN	O	O
at	NN	O	O
different	NN	O	O
sites	NN	O	O
within	NN	O	O
this	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
abolished	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
cooperation	NN	O	O
of	NN	O	O
different	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
IL-9	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

DNA	NN	O	O
triplex	NN	O	O
formation	NN	O	O
selectively	NN	O	O
inhibits	NN	O	O
granulocyte-macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
human	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

Granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
hemopoietic	NN	O	O
growth	NN	O	O
factor	NN	O	O
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
activated	NN	O	O
T	NN	O	O
cells	NN	O	O
,	NN	O	O
fibroblasts	NN	O	O
,	NN	O	O
macrophages	NN	O	O
,	NN	O	O
and	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
.	NN	O	O

Although	NN	O	O
GM-CSF	NN	O	B-protein
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
essential	NN	O	O
for	NN	O	O
normal	NN	O	O
hemopoiesis	NN	O	O
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
GM-CSF	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
some	NN	O	O
diseases	NN	O	O
such	NN	O	O
as	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
and	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
.	NN	O	O

An	NN	O	O
NF-kappaB	NN	O	B-protein
/Rel	NN	O	O
binding	NN	O	O
site	NN	O	O
within	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
termed	NN	O	O
the	NN	O	O
kappaB	NN	O	O
element	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
controlling	NN	O	O
expression	NN	O	O
in	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
assays	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
stimuli	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
oligonucleotide-directed	NN	O	O
triple	NN	O	O
helix	NN	O	O
formation	NN	O	O
across	NN	O	O
this	NN	O	O
regulatory	NN	O	B-DNA
sequence	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
potential	NN	O	O
tool	NN	O	O
to	NN	O	O
inhibit	NN	O	O
GM-CSF	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

A	NN	O	O
15-base	NN	O	O
oligonucleotide	NN	O	O
,	NN	O	O
GM3	NN	O	O
,	NN	O	O
was	NN	O	O
targeted	NN	O	O
to	NN	O	O
a	NN	O	O
purine-rich	NN	O	B-DNA
region	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
proximal	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
overlaps	NN	O	O
the	NN	O	O
kappaB	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

Gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
and	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
footprinting	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
GM3	NN	O	O
formed	NN	O	O
a	NN	O	O
sequence-specific	NN	O	O
collinear	NN	O	O
triplex	NN	O	O
with	NN	O	O
its	NN	O	O
double-stranded	NN	O	B-DNA
DNA	NN	O	I-DNA
target	NN	O	I-DNA
.	NN	O	O

Triplex	NN	O	O
formation	NN	O	O
by	NN	O	O
GM3	NN	O	O
blocked	NN	O	O
recombinant	NN	O	B-protein
and	NN	O	I-protein
nuclear	NN	O	I-protein
NF-kappaB	NN	O	I-protein
proteins	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

GM3	NN	O	O
also	NN	O	O
caused	NN	O	O
selective	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
lymphotrophic	NN	O	I-protein
virus-1	NN	O	I-protein
Tax	NN	O	I-protein
transactivator-induced	NN	O	O
luciferase	NN	O	B-protein
activity	NN	O	O
from	NN	O	O
a	NN	O	O
reporter	NN	O	O
construct	NN	O	O
driven	NN	O	O
by	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
GM3	NN	O	O
greatly	NN	O	O
reduced	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
endogenous	NN	O	O
GM-CSF	NN	O	B-RNA
mRNA	NN	O	I-RNA
induced	NN	O	O
by	NN	O	O
different	NN	O	O
stimuli	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
interleukin	NN	O	B-RNA
3	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
cells	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
kappaB	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
plays	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Colinear	NN	O	O
triplex	NN	O	O
formation	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
selective	NN	O	O
transcriptional	NN	O	O
repressor	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
may	NN	O	O
have	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
application	NN	O	O
in	NN	O	O
cases	NN	O	O
of	NN	O	O
undesirable	NN	O	O
overexpression	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reversible	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
monoblastic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
ML-9	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
myosin	NN	O	B-protein
light	NN	O	I-protein
chain	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

Human	NN	O	B-cell_line
monoblastic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
into	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
by	NN	O	O
various	NN	O	O
agents	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
1-	NN	O	O
(	NN	O	O
5-chloronaphthalene-1-sulfonyl	NN	O	O
)	NN	O	O
-1H-hexahydro-1	NN	O	O
,	NN	O	O
4-diazepine	NN	O	O
hydrochloride	NN	O	O
(	NN	O	O
ML-9	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
myosin	NN	O	B-protein
light	NN	O	I-protein
chain	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
induces	NN	O	O
differentiation	NN	O	O
of	NN	O	O
monocytoid	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
U937	NN	O	B-cell_line
and	NN	O	O
THP-1	NN	O	B-cell_line
but	NN	O	O
not	NN	O	O
of	NN	O	O
myeloblastic	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
ML-1	NN	O	I-cell_line
cell	NN	O	I-cell_line
or	NN	O	O
erythroleukemia	NN	O	B-cell_line
K562	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
further	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
ML-9	NN	O	O
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
VD3	NN	O	O
)	NN	O	O
a	NN	O	O
typical	NN	O	O
inducer	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

ML-9	NN	O	O
induced	NN	O	O
nitroblue	NN	O	O
tetrazolium	NN	O	O
(	NN	O	O
NBT	NN	O	O
)	NN	O	O
-reducing	NN	O	O
activity	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cell	NN	O	I-cell_line
more	NN	O	O
rapidly	NN	O	O
than	NN	O	O
VD3	NN	O	O
:	NN	O	O
This	NN	O	O
differentiation	NN	O	O
marker	NN	O	O
was	NN	O	O
induced	NN	O	O
significantly	NN	O	O
after	NN	O	O
incubation	NN	O	O
with	NN	O	O
ML-9	NN	O	O
and	NN	O	O
VD3	NN	O	O
for	NN	O	O
4	NN	O	O
hours	NN	O	O
and	NN	O	O
1	NN	O	O
day	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

ML-9	NN	O	O
also	NN	O	O
induced	NN	O	O
alpha-naphthyl	NN	O	B-protein
acetate	NN	O	I-protein
esterase	NN	O	I-protein
(	NN	O	O
ANAE	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
,	NN	O	O
another	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
marker	NN	O	O
,	NN	O	O
more	NN	O	O
rapidly	NN	O	O
than	NN	O	O
VD3	NN	O	O
.	NN	O	O

The	NN	O	O
maximum	NN	O	O
levels	NN	O	O
of	NN	O	O
these	NN	O	O
markers	NN	O	O
induced	NN	O	O
by	NN	O	O
ML-9	NN	O	O
were	NN	O	O
comparable	NN	O	O
to	NN	O	O
those	NN	O	O
induced	NN	O	O
by	NN	O	O
VD3	NN	O	O
,	NN	O	O
but	NN	O	O
after	NN	O	O
removal	NN	O	O
of	NN	O	O
ML-9	NN	O	O
from	NN	O	O
the	NN	O	O
medium	NN	O	O
by	NN	O	O
washing	NN	O	O
the	NN	O	O
cells	NN	O	O
,	NN	O	O
the	NN	O	O
expressions	NN	O	O
of	NN	O	O
theses	NN	O	O
markers	NN	O	O
decreased	NN	O	O
within	NN	O	O
4	NN	O	O
hours	NN	O	O
and	NN	O	O
reached	NN	O	O
basal	NN	O	O
levels	NN	O	O
in	NN	O	O
1	NN	O	O
day	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
ML-9	NN	O	O
's	NN	O	O
induction	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
differentiation-associated	NN	O	O
phenotypes	NN	O	O
was	NN	O	O
reversible	NN	O	O
.	NN	O	O

The	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
ML-9	NN	O	O
was	NN	O	O
also	NN	O	O
reversible	NN	O	O
.	NN	O	O

Similar	NN	O	O
effects	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
another	NN	O	O
line	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
monoblastic	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
.	NN	O	O

ML-9	NN	O	O
had	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
morphology	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
increased	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
monocyte-macrophage	NN	O	B-protein
lineage-associated	NN	O	I-protein
surface	NN	O	I-protein
antigen	NN	O	I-protein
,	NN	O	O
CD14	NN	O	B-protein
,	NN	O	O
to	NN	O	O
some	NN	O	O
extent	NN	O	O
.	NN	O	O

Irreversible	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
VD3	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
down	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-myc	NN	O	O
and	NN	O	O
upregulation	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	O
and	NN	O	O
c-jun	NN	O	O
,	NN	O	O
but	NN	O	O
ML-9	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
oncogenes	NN	O	O
appreciably	NN	O	O
.	NN	O	O

ML-9-induced	NN	O	O
differentiation	NN	O	O
was	NN	O	O
also	NN	O	O
reversible	NN	O	O
when	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
cultured	NN	O	O
with	NN	O	O
cultured	NN	O	O
with	NN	O	O
ML-9	NN	O	O
plus	NN	O	O
an	NN	O	O
anti-cancer	NN	O	O
drug	NN	O	O
such	NN	O	O
as	NN	O	O
1-beta-D-arabino-furanosylcytosine	NN	O	O
or	NN	O	O
daunomycin	NN	O	O
.	NN	O	O

it	NN	O	O
became	NN	O	O
irreversible	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
upon	NN	O	O
simultaneous	NN	O	O
treatment	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
TGF-beta	NN	O	B-protein
1	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
differentiation	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
caused	NN	O	O
growth	NN	O	O
arrest	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
G0/G1	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
ML-9	NN	O	O
should	NN	O	O
be	NN	O	O
useful	NN	O	O
for	NN	O	O
studying	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Modulation	NN	O	O
of	NN	O	O
endogenous	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	I-protein
antagonist	NN	O	O
results	NN	O	O
in	NN	O	O
opposing	NN	O	O
effects	NN	O	O
on	NN	O	O
HIV	NN	O	O
expression	NN	O	O
in	NN	O	O
chronically	NN	O	B-cell_line
infected	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
proportion	NN	O	O
of	NN	O	O
HIV-infected	NN	O	O
individuals	NN	O	O
experience	NN	O	O
episodes	NN	O	O
of	NN	O	O
localized	NN	O	O
or	NN	O	O
systemic	NN	O	O
bacterial	NN	O	O
infections	NN	O	O
caused	NN	O	O
by	NN	O	O
Gram-negative	NN	O	O
bacteria	NN	O	O
.	NN	O	O

Many	NN	O	O
of	NN	O	O
the	NN	O	O
clinical	NN	O	O
side	NN	O	O
effects	NN	O	O
of	NN	O	O
these	NN	O	O
infections	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
which	NN	O	O
are	NN	O	O
induced	NN	O	O
primarily	NN	O	O
by	NN	O	O
LPS	NN	O	O
,	NN	O	O
a	NN	O	O
constituent	NN	O	O
of	NN	O	O
the	NN	O	O
bacterial	NN	O	O
cell	NN	O	O
wall	NN	O	O
of	NN	O	O
Gram-negative	NN	O	O
bacteria	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
examines	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
LPS-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
in	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
promonocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
chronically	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
LPS	NN	O	O
alone	NN	O	O
induced	NN	O	O
minimal	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
by	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Costimulation	NN	O	O
of	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
LPS	NN	O	O
plus	NN	O	O
GM-CSF	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
steady-state	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV	NN	O	B-RNA
RNA	NN	O	I-RNA
;	NN	O	O
however	NN	O	O
,	NN	O	O
only	NN	O	O
a	NN	O	O
weak	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
-driven	NN	O	O
transcription	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
was	NN	O	O
noted	NN	O	O
.	NN	O	O

Costimulation	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
LPS	NN	O	O
plus	NN	O	O
GM-CSF	NN	O	B-protein
induced	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

IL-1	NN	O	B-protein
receptor	NN	O	I-protein
antagonist	NN	O	O
(	NN	O	O
ra	NN	O	O
)	NN	O	O
inhibited	NN	O	O
LPS	NN	O	O
enhancement	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
in	NN	O	O
GM-CSF-stimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
endogenous	NN	O	B-protein
IL-1	NN	O	I-protein
was	NN	O	O
involved	NN	O	O
in	NN	O	O
LPS-mediated	NN	O	O
viral	NN	O	O
production	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
regard	NN	O	O
,	NN	O	O
anti-inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
inhibited	NN	O	O
LPS	NN	O	O
plus	NN	O	O
GM-CSF	NN	O	B-protein
-stimulated	NN	O	O
HIV	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
effect	NN	O	O
closely	NN	O	O
correlated	NN	O	O
with	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
release	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
with	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
endogenous	NN	O	O
IL-1ra	NN	O	O
production	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
balance	NN	O	O
between	NN	O	O
an	NN	O	O
endogenously	NN	O	B-protein
produced	NN	O	I-protein
viral	NN	O	I-protein
inducer	NN	O	I-protein
(	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
)	NN	O	O
and	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
IL-1ra	NN	O	B-protein
)	NN	O	O
may	NN	O	O
represent	NN	O	O
an	NN	O	O
important	NN	O	O
pathway	NN	O	O
leading	NN	O	O
to	NN	O	O
modulation	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
from	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Herpesvirus	NN	O	B-cell_line
saimiri	NN	O	I-cell_line
immortalized	NN	O	I-cell_line
gamma	NN	O	I-cell_line
delta	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
activated	NN	O	O
by	NN	O	O
IL-12	NN	O	B-protein
.	NN	O	O

IL-12	NN	O	B-protein
is	NN	O	O
a	NN	O	O
novel	NN	O	O
heterodimeric	NN	O	B-protein
cytokine	NN	O	I-protein
important	NN	O	O
for	NN	O	O
the	NN	O	O
regulation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
lymphocytes	NN	O	O
and	NN	O	O
NK	NN	O	O
cells	NN	O	O
.	NN	O	O

Like	NN	O	O
other	NN	O	O
cytokines	NN	O	O
,	NN	O	O
IL-12	NN	O	B-protein
mediates	NN	O	O
its	NN	O	O
biologic	NN	O	O
activity	NN	O	O
through	NN	O	O
high-affinity	NN	O	O
receptors	NN	O	O
expressed	NN	O	O
on	NN	O	O
responsive	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
date	NN	O	O
,	NN	O	O
a	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
receptors	NN	O	O
for	NN	O	O
IL-12	NN	O	B-protein
have	NN	O	O
been	NN	O	O
found	NN	O	O
only	NN	O	O
on	NN	O	O
PBMC	NN	O	B-cell_type
following	NN	O	O
activation	NN	O	O
with	NN	O	O
PHA	NN	O	B-protein
or	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

To	NN	O	O
gain	NN	O	O
further	NN	O	O
knowledge	NN	O	O
of	NN	O	O
the	NN	O	O
IL-12R	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
the	NN	O	O
IL-12	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
in	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
studied	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
had	NN	O	O
been	NN	O	O
transformed	NN	O	O
to	NN	O	O
permanent	NN	O	O
growth	NN	O	O
with	NN	O	O
Herpesvirus	NN	O	O
saimiri	NN	O	O
,	NN	O	O
an	NN	O	O
oncogenic	NN	O	O
virus	NN	O	O
of	NN	O	O
nonhuman	NN	O	O
primates	NN	O	O
.	NN	O	O

This	NN	O	O
paper	NN	O	O
reports	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-12R	NN	O	B-protein
on	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
gamma	NN	O	I-cell_line
delta	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
that	NN	O	O
responds	NN	O	O
to	NN	O	O
IL-12	NN	O	B-protein
with	NN	O	O
enhanced	NN	O	O
cytolytic	NN	O	O
activity	NN	O	O
and	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
cytolytic	NN	O	B-protein
effector	NN	O	I-protein
molecules	NN	O	I-protein
granzyme	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
perforin	NN	O	B-protein
.	NN	O	O

Using	NN	O	O
these	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
a	NN	O	O
model	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
,	NN	O	O
we	NN	O	O
confirmed	NN	O	O
that	NN	O	O
these	NN	O	O
events	NN	O	O
involve	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
nonreceptor	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
JAK2	NN	O	B-protein
,	NN	O	O
TYK2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
4	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
at	NN	O	O
early	NN	O	O
stages	NN	O	O
of	NN	O	O
B-cell	NN	O	O
and	NN	O	O
T-cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
distinct	NN	O	O
sets	NN	O	O
of	NN	O	O
genes	NN	O	O
at	NN	O	O
different	NN	O	O
stages	NN	O	O
of	NN	O	O
B-lymphocyte	NN	O	O
and	NN	O	O
T-lymphocyte	NN	O	O
differentiation	NN	O	O
is	NN	O	O
controlled	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

A	NN	O	O
number	NN	O	O
of	NN	O	O
recent	NN	O	O
studies	NN	O	O
have	NN	O	O
described	NN	O	O
interactions	NN	O	O
between	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	O
that	NN	O	O
provide	NN	O	O
new	NN	O	O
insights	NN	O	O
into	NN	O	O
mechanisms	NN	O	O
regulating	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

These	NN	O	O
mechanisms	NN	O	O
include	NN	O	O
the	NN	O	O
assembly	NN	O	O
of	NN	O	O
higher	NN	O	B-protein
order	NN	O	I-protein
nucleoprotein	NN	O	I-protein
complexes	NN	O	I-protein
and	NN	O	O
other	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
that	NN	O	O
enhance	NN	O	O
the	NN	O	O
functional	NN	O	O
specificity	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
regulators	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
LIM-Hox	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
hLH-2	NN	O	B-DNA
,	NN	O	O
aberrantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukaemia	NN	O	O
and	NN	O	O
located	NN	O	O
on	NN	O	O
9q33-34.1	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
describe	NN	O	O
the	NN	O	O
isolation	NN	O	O
of	NN	O	O
human	NN	O	B-protein
LH-2	NN	O	I-protein
,	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
containing	NN	O	O
two	NN	O	O
cysteine-rich	NN	O	B-protein
regions	NN	O	I-protein
(	NN	O	O
LIM	NN	O	B-protein
domains	NN	O	I-protein
)	NN	O	O
and	NN	O	O
a	NN	O	O
homeobox	NN	O	B-protein
(	NN	O	I-protein
Hox	NN	O	I-protein
)	NN	O	I-protein
DNA-binding	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

High	NN	O	O
levels	NN	O	O
of	NN	O	O
hLH-2	NN	O	B-DNA
expression	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
all	NN	O	O
cases	NN	O	O
of	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukaemia	NN	O	O
(	NN	O	O
CML	NN	O	O
)	NN	O	O
tested	NN	O	O
,	NN	O	O
regardless	NN	O	O
of	NN	O	O
disease	NN	O	O
status	NN	O	O
.	NN	O	O

hLH-2	NN	O	B-DNA
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
chromosome	NN	O	B-DNA
9Q33-34.1	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
region	NN	O	O
as	NN	O	O
the	NN	O	O
reciprocal	NN	O	O
translocation	NN	O	O
that	NN	O	O
creates	NN	O	O
the	NN	O	O
BCR-ABL	NN	O	B-protein
chimera	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Philadelphia	NN	O	O
chromosome	NN	O	O
(	NN	O	O
Ph	NN	O	O
'	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
hallmark	NN	O	O
of	NN	O	O
CML	NN	O	O
;	NN	O	O
hLH-2	NN	O	B-DNA
was	NN	O	O
retained	NN	O	O
on	NN	O	O
the	NN	O	O
derivative	NN	O	B-DNA
9	NN	O	I-DNA
chromosome	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
therefore	NN	O	O
centromeric	NN	O	O
of	NN	O	O
c-ABL	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
proximity	NN	O	O
of	NN	O	O
hLH-2	NN	O	B-DNA
to	NN	O	O
the	NN	O	O
breakpoint	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
9	NN	O	I-DNA
raises	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
cis-activation	NN	O	O
by	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
9	NN	O	I-DNA
;	NN	O	I-DNA
22	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q34	NN	O	I-DNA
;	NN	O	I-DNA
q11	NN	O	I-DNA
)	NN	O	I-DNA
translocation	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
finding	NN	O	O
hLH-2	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
all	NN	O	O
cases	NN	O	O
of	NN	O	O
CML	NN	O	O
,	NN	O	O
expression	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
malignancies	NN	O	O
and	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
primary	NN	O	O
cases	NN	O	O
of	NN	O	O
acute	NN	O	O
myelogenous	NN	O	O
leukaemia	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
hLH-2	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
development	NN	O	O
or	NN	O	O
progression	NN	O	O
of	NN	O	O
leukaemia	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
hLH-2	NN	O	B-DNA
may	NN	O	O
prove	NN	O	O
useful	NN	O	O
as	NN	O	O
a	NN	O	O
marker	NN	O	O
of	NN	O	O
CML	NN	O	O
for	NN	O	O
monitoring	NN	O	O
residual	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lymphoid	NN	O	O
cell	NN	O	O
resistance	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
HIV	NN	O	O
infection	NN	O	O
.	NN	O	O

In	NN	O	O
humans	NN	O	O
infected	NN	O	O
with	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
virus	NN	O	O
there	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
disproportionate	NN	O	O
severity	NN	O	O
of	NN	O	O
signs	NN	O	O
and	NN	O	O
symptoms	NN	O	O
of	NN	O	O
illness	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
fraction	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_line
infected	NN	O	I-cell_line
T-lymphoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
part	NN	O	O
,	NN	O	O
this	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
altered	NN	O	O
intercellular	NN	O	O
signalling	NN	O	O
systems	NN	O	O
and	NN	O	O
intracellular	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
are	NN	O	O
well	NN	O	O
known	NN	O	O
for	NN	O	O
their	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
vitality	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
HIV	NN	O	O
infections	NN	O	O
often	NN	O	O
show	NN	O	O
elevated	NN	O	O
circulating	NN	O	O
levels	NN	O	O
of	NN	O	O
cortisol	NN	O	O
,	NN	O	O
suggesting	NN	O	O
some	NN	O	O
misfunction	NN	O	O
in	NN	O	O
the	NN	O	O
regulatory	NN	O	O
systems	NN	O	O
that	NN	O	O
maintain	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
this	NN	O	O
critical	NN	O	O
hormone	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
cellular	NN	O	O
level	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
known	NN	O	O
that	NN	O	O
both	NN	O	O
acute	NN	O	O
HIV	NN	O	O
infection	NN	O	O
and	NN	O	O
glucocorticoids	NN	O	O
can	NN	O	O
cause	NN	O	O
apoptotic	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
thymic	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
chronically	NN	O	B-cell_type
HIV-infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
appear	NN	O	O
to	NN	O	O
be	NN	O	O
resistant	NN	O	O
to	NN	O	O
glucocorticoid-evoked	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
-ligand	NN	O	O
binding	NN	O	O
studies	NN	O	O
on	NN	O	O
patients	NN	O	O
'	NN	O	O
cells	NN	O	O
have	NN	O	O
shown	NN	O	O
reduced	NN	O	O
affinity	NN	O	O
between	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
and	NN	O	O
test	NN	O	O
steroids	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
,	NN	O	O
chronically	NN	O	B-cell_line
HIV-infected	NN	O	I-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
lymphoid	NN	O	B-cell_line
CEM	NN	O	I-cell_line
line	NN	O	I-cell_line
displayed	NN	O	O
resistance	NN	O	O
to	NN	O	O
glucocorticoid-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

These	NN	O	O
cells	NN	O	O
showed	NN	O	O
reduced	NN	O	O
numbers	NN	O	O
of	NN	O	O
binding	NN	O	B-protein
sites	NN	O	I-protein
with	NN	O	O
little	NN	O	O
alteration	NN	O	O
in	NN	O	O
apparent	NN	O	O
affinity	NN	O	O
between	NN	O	O
ligand	NN	O	O
and	NN	O	O
receptor	NN	O	O
.	NN	O	O

Thus	NN	O	O
it	NN	O	O
appears	NN	O	O
that	NN	O	O
there	NN	O	O
may	NN	O	O
often	NN	O	O
be	NN	O	O
malfunction	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
glucocorticoid	NN	O	O
response	NN	O	O
in	NN	O	O
HIV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
probably	NN	O	O
due	NN	O	O
to	NN	O	O
altered	NN	O	O
interactions	NN	O	O
between	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
its	NN	O	O
hormone	NN	O	O
.	NN	O	O

Such	NN	O	O
alterations	NN	O	O
may	NN	O	O
have	NN	O	O
clinical	NN	O	O
consequences	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
relatively	NN	O	O
longer	NN	O	O
life	NN	O	O
span	NN	O	O
of	NN	O	O
infected	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
systemic	NN	O	O
effects	NN	O	O
of	NN	O	O
chronically	NN	O	O
elevated	NN	O	O
cortisol	NN	O	O
level	NN	O	O
.	NN	O	O

-DOCSTART-	O

Abundant	NN	O	O
expression	NN	O	O
of	NN	O	O
erythroid	NN	O	B-RNA
transcription	NN	O	I-RNA
factor	NN	O	I-RNA
P45	NN	O	I-RNA
NF-E2	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
granurocytes	NN	O	I-cell_type
.	NN	O	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-E2	NN	O	I-protein
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
regulation	NN	O	O
of	NN	O	O
erythroid-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

p45	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-E2	NN	O	B-protein
contains	NN	O	O
a	NN	O	O
basic-leucine	NN	O	B-protein
zipper	NN	O	I-protein
domain	NN	O	I-protein
and	NN	O	O
dimerizes	NN	O	O
with	NN	O	O
the	NN	O	O
small	NN	O	B-protein
Maf	NN	O	I-protein
family	NN	O	I-protein
protein	NN	O	I-protein
to	NN	O	O
form	NN	O	O
functional	NN	O	O
NF-E2	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

While	NN	O	O
p45	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
restricted	NN	O	O
to	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
megakaryocytes	NN	O	B-cell_type
and	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
hematopoietic	NN	O	O
lineage	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
that	NN	O	O
p45	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
abundantly	NN	O	O
transcribed	NN	O	O
in	NN	O	O
the	NN	O	O
granulocyte	NN	O	B-cell_type
fraction	NN	O	I-cell_type
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

As	NN	O	O
neutrophils	NN	O	B-cell_type
occupy	NN	O	O
approximately	NN	O	O
92	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
in	NN	O	O
granulocyte	NN	O	B-cell_type
fraction	NN	O	I-cell_type
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

As	NN	O	O
neutrophils	NN	O	B-cell_type
occupy	NN	O	O
approximately	NN	O	O
92	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
in	NN	O	O
this	NN	O	O
fraction	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
expressing	NN	O	O
p45	NN	O	B-protein
is	NN	O	O
most	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
neutrophils	NN	O	B-cell_type
.	NN	O	O

p45	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
also	NN	O	O
expressed	NN	O	O
in	NN	O	O
HL-60	NN	O	B-cell_line
promyelocytes	NN	O	I-cell_line
,	NN	O	O
albeit	NN	O	O
the	NN	O	O
expression	NN	O	O
level	NN	O	O
is	NN	O	O
much	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
granulocyte	NN	O	B-cell_type
fraction	NN	O	I-cell_type
.	NN	O	O

HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
found	NN	O	O
to	NN	O	O
express	NN	O	O
mafK	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
genuine	NN	O	O
NF-E2	NN	O	B-protein
complex	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

Although	NN	O	O
p45	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
transcribed	NN	O	O
from	NN	O	O
two	NN	O	O
different	NN	O	O
promoters	NN	O	O
,	NN	O	O
aNF-E2	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
fNF-E2	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
p45	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
transcribed	NN	O	O
only	NN	O	O
from	NN	O	O
aNF-E2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
p45	NN	O	B-protein
megakaryocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
p45	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
transcribed	NN	O	O
only	NN	O	O
from	NN	O	O
aNF-E2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
p45	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
neutrophils	NN	O	B-cell_type
declined	NN	O	O
rapidly	NN	O	O
after	NN	O	O
transfer	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
to	NN	O	O
in	NN	O	O
vitro	NN	O	O
culture	NN	O	O
and	NN	O	O
G-CSF	NN	O	B-protein
could	NN	O	O
not	NN	O	O
sustain	NN	O	O
the	NN	O	O
expression	NN	O	O
from	NN	O	O
the	NN	O	O
down-regulation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
E2	NN	O	B-protein
may	NN	O	O
also	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
neutrophil-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-protein
and	NN	O	I-protein
c-jun	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
during	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
phytohemagglutinin-stimulated	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
protein	NN	O	O
products	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
(	NN	O	O
p62c-fos	NN	O	B-protein
)	NN	O	O
and	NN	O	O
c-jun	NN	O	B-DNA
(	NN	O	O
p39c-jun	NN	O	B-protein
)	NN	O	O
genes	NN	O	O
are	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
and	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
play	NN	O	O
important	NN	O	O
roles	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

Most	NN	O	O
studies	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
have	NN	O	O
been	NN	O	O
performed	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
,	NN	O	O
and	NN	O	O
therefore	NN	O	O
do	NN	O	O
not	NN	O	O
give	NN	O	O
direct	NN	O	O
information	NN	O	O
about	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
proteins	NN	O	O
during	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
Western	NN	O	O
blotting	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
during	NN	O	O
the	NN	O	O
cell	NN	O	O
cycles	NN	O	O
of	NN	O	O
two	NN	O	O
different	NN	O	O
cellular	NN	O	B-cell_line
systems	NN	O	I-cell_line
:	NN	O	O
a	NN	O	O
continuously	NN	O	B-cell_line
growing	NN	O	I-cell_line
myeloid	NN	O	I-cell_line
leukemic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
HL60	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
into	NN	O	O
cycle	NN	O	O
,	NN	O	O
phyto-	NN	O	B-cell_line
hemagglutinin	NN	O	I-cell_line
(	NN	O	I-cell_line
PHA	NN	O	I-cell_line
)	NN	O	I-cell_line
-stimulated	NN	O	I-cell_line
normal	NN	O	I-cell_line
human	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
blood	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
(	NN	O	O
PBL	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
transcription	NN	O	O
factor	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
consists	NN	O	O
of	NN	O	O
dimers	NN	O	O
of	NN	O	O
Fos	NN	O	B-protein
and	NN	O	I-protein
Jun	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
was	NN	O	O
also	NN	O	O
studied	NN	O	O
using	NN	O	O
a	NN	O	O
gel	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
nuclear	NN	O	O
p62c-fos	NN	O	B-protein
,	NN	O	O
p39c-jun	NN	O	B-protein
,	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
throughout	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
both	NN	O	O
in	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
PHA-stimulated	NN	O	B-cell_line
PBL	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
we	NN	O	O
postulate	NN	O	O
that	NN	O	O
these	NN	O	O
proteins	NN	O	O
are	NN	O	O
required	NN	O	O
throughout	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
and	NN	O	O
not	NN	O	O
transiently	NN	O	O
in	NN	O	O
the	NN	O	O
G0	NN	O	O
to	NN	O	O
G1	NN	O	O
transition	NN	O	O
as	NN	O	O
previous	NN	O	O
mRNA	NN	O	O
studies	NN	O	O
have	NN	O	O
indicated	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
an	NN	O	O
uncoupling	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
from	NN	O	O
p62c-fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p39c-jun	NN	O	B-protein
AP-1	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
expressed	NN	O	O
more	NN	O	O
strongly	NN	O	O
in	NN	O	O
the	NN	O	O
G1-	NN	O	O
and	NN	O	O
G2/M-phase	NN	O	O
enriched	NN	O	O
samples	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
S-phase	NN	O	O
enriched	NN	O	O
samples	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
levels	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
p62c-fos	NN	O	I-protein
and	NN	O	I-protein
p39c-jun	NN	O	I-protein
were	NN	O	O
constant	NN	O	O
.	NN	O	O

Nuclei	NN	O	O
of	NN	O	O
unstimulated	NN	O	B-cell_type
PBL	NN	O	I-cell_type
from	NN	O	O
different	NN	O	O
donors	NN	O	O
expressed	NN	O	O
p62c-fos	NN	O	B-protein
and	NN	O	O
p39c-jun	NN	O	B-protein
,	NN	O	O
but	NN	O	O
AP-1	NN	O	B-protein
was	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
samples	NN	O	O
.	NN	O	O

Following	NN	O	O
PHA	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
PBL	NN	O	B-cell_type
,	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
delayed	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
augmentation	NN	O	O
of	NN	O	O
p39c-jun	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
observed	NN	O	O
that	NN	O	O
cytoplasmic	NN	O	B-protein
p62c-fos	NN	O	I-protein
and	NN	O	I-protein
p39c-jun	NN	O	I-protein
were	NN	O	O
present	NN	O	O
in	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
PHA-stimulated	NN	O	B-cell_line
PBL	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
no	NN	O	O
cytoplasmic	NN	O	B-protein
p62c-fos	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
unstimulated	NN	O	B-cell_type
PBL	NN	O	I-cell_type
,	NN	O	O
although	NN	O	O
in	NN	O	O
some	NN	O	O
cases	NN	O	O
cytoplasmic	NN	O	B-protein
p39c-jun	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
subcellular	NN	O	O
compartmentalization	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	B-protein
may	NN	O	O
occur	NN	O	O
under	NN	O	O
certain	NN	O	O
circumstances	NN	O	O
.	NN	O	O

-DOCSTART-	O

Octamer	NN	O	B-protein
binding	NN	O	I-protein
factors	NN	O	I-protein
and	NN	O	O
their	NN	O	O
coactivator	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
PU.1	NN	O	I-DNA
(	NN	O	I-DNA
spi-1	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

PU.1	NN	O	B-protein
(	NN	O	O
spi-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
is	NN	O	O
predominantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
activates	NN	O	O
many	NN	O	O
B	NN	O	B-DNA
cell	NN	O	I-DNA
and	NN	O	I-DNA
myeloid	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
development	NN	O	O
of	NN	O	O
both	NN	O	O
of	NN	O	O
these	NN	O	O
lineages	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
studies	NN	O	O
(	NN	O	O
Chen	NN	O	O
,	NN	O	O
H.M.	NN	O	O
,	NN	O	O
Ray-Gallet	NN	O	O
,	NN	O	O
D.	NN	O	O
,	NN	O	O
Zhang	NN	O	O
,	NN	O	O
P.	NN	O	O
,	NN	O	O
Hetherington	NN	O	O
,	NN	O	O
C.J.	NN	O	O
,	NN	O	O
Gonzalez	NN	O	O
,	NN	O	O
D.A.	NN	O	O
,	NN	O	O
Zhang	NN	O	O
,	NN	O	O
D.-E.	NN	O	O
,	NN	O	O
Moreau-Gachelin	NN	O	O
,	NN	O	O
F.	NN	O	O
,	NN	O	O
and	NN	O	O
Tenen	NN	O	O
,	NN	O	O
D.G.	NN	O	O
(	NN	O	O
1995	NN	O	O
)	NN	O	O
Oncogene	NN	O	O
11	NN	O	O
,	NN	O	O
1549-1560	NN	O	O
)	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
PU.1	NN	O	B-DNA
promoter	NN	O	I-DNA
directs	NN	O	O
cell	NN	O	B-DNA
type-specific	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
lines	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
PU.1	NN	O	B-protein
activates	NN	O	O
its	NN	O	O
own	NN	O	O
promoter	NN	O	O
in	NN	O	O
an	NN	O	O
autoregulatory	NN	O	O
loop	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
PU.1	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
also	NN	O	O
specifically	NN	O	O
and	NN	O	O
highly	NN	O	O
functional	NN	O	O
in	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
.	NN	O	O

Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
can	NN	O	O
bind	NN	O	O
specifically	NN	O	O
to	NN	O	O
a	NN	O	O
site	NN	O	O
at	NN	O	O
base	NN	O	O
pair	NN	O	O
-55	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
site	NN	O	O
is	NN	O	O
specifically	NN	O	O
protected	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
two	NN	O	O
other	NN	O	O
sites	NN	O	O
contribute	NN	O	O
to	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
an	NN	O	O
Sp1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
adjacent	NN	O	O
to	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
PU.1	NN	O	B-DNA
autoregulatory	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
B	NN	O	O
cell	NN	O	O
coactivator	NN	O	B-protein
OBF-1/Bob1/OCA-B	NN	O	B-protein
is	NN	O	O
only	NN	O	O
expressed	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
not	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
that	NN	O	O
OBF-1/Bob1/OCA-B	NN	O	B-protein
can	NN	O	O
transactivate	NN	O	O
the	NN	O	O
PU.1	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
HeLa	NN	O	B-cell_line
and	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
restricted	NN	O	I-protein
coactivator	NN	O	I-protein
may	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
B	NN	O	O
cell	NN	O	O
specific	NN	O	O
expression	NN	O	O
of	NN	O	O
PU.1	NN	O	B-protein
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
processing	NN	O	O
by	NN	O	O
NF-kappaB	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
transiently	NN	O	B-cell_line
transfected	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
NF-kappaB	NN	O	B-protein
involves	NN	O	O
proteasome-mediated	NN	O	O
processing	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB1	NN	O	B-protein
p105	NN	O	I-protein
precursor	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
generates	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
processing	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
occurs	NN	O	O
constitutively	NN	O	O
in	NN	O	O
vivo	NN	O	O
but	NN	O	O
can	NN	O	O
be	NN	O	O
markedly	NN	O	O
enhanced	NN	O	O
by	NN	O	O
various	NN	O	O
cellular	NN	O	O
activation	NN	O	O
agents	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
underlying	NN	O	O
regulatory	NN	O	O
mechanism	NN	O	O
is	NN	O	O
not	NN	O	O
yet	NN	O	O
clear	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
signal-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
processing	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
de	NN	O	O
novo	NN	O	O
synthesis	NN	O	O
of	NN	O	O
this	NN	O	O
precursor	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
studies	NN	O	O
performed	NN	O	O
in	NN	O	O
COS7	NN	O	B-cell_line
cells	NN	O	I-cell_line
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
newly	NN	O	O
synthesized	NN	O	O
p105	NN	O	B-protein
protein	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
more	NN	O	O
rapidly	NN	O	O
processed	NN	O	O
compared	NN	O	O
to	NN	O	O
its	NN	O	O
accumulated	NN	O	O
form	NN	O	O
that	NN	O	O
is	NN	O	O
already	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
processed	NN	O	O
product	NN	O	O
p50	NN	O	B-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
processing	NN	O	O
rate	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
is	NN	O	O
markedly	NN	O	O
inhibited	NN	O	O
in	NN	O	O
cells	NN	O	O
co-transfected	NN	O	O
with	NN	O	O
p50	NN	O	B-protein
or	NN	O	O
other	NN	O	O
NF-kappaB	NN	O	B-protein
subunits	NN	O	I-protein
,	NN	O	O
including	NN	O	O
RelA	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
that	NN	O	O
physically	NN	O	O
interact	NN	O	O
with	NN	O	O
p105	NN	O	B-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
processing	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
is	NN	O	O
subject	NN	O	O
to	NN	O	O
negative	NN	O	O
regulation	NN	O	O
by	NN	O	O
the	NN	O	O
various	NN	O	O
NF-kappaB	NN	O	B-protein
subunits	NN	O	I-protein
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
p105	NN	O	B-protein
undergoes	NN	O	O
degradation	NN	O	O
in	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-cell_line
human	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
inducible	NN	O	O
degradation	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
is	NN	O	O
not	NN	O	O
coupled	NN	O	O
with	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
processing	NN	O	O
and	NN	O	O
inducible	NN	O	O
degradation	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
are	NN	O	O
differentially	NN	O	O
regulated	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	B-protein
regulator	NN	O	I-protein
of	NN	O	O
c-myb	NN	O	B-DNA
expression	NN	O	O
during	NN	O	O
T-cell	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

Resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
very	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
c-Myb	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

During	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
c-myb	NN	O	B-DNA
expression	NN	O	O
is	NN	O	O
induced	NN	O	O
and	NN	O	O
much	NN	O	O
of	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
expression	NN	O	O
occurs	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
c-myb	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
sequence	NN	O	I-DNA
that	NN	O	O
increased	NN	O	O
c-myb	NN	O	B-DNA
expression	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
footprinting	NN	O	O
by	NN	O	O
ligation-mediated	NN	O	O
PCR	NN	O	O
was	NN	O	O
performed	NN	O	O
to	NN	O	O
correlate	NN	O	O
in	NN	O	O
vivo	NN	O	O
protein	NN	O	O
binding	NN	O	O
with	NN	O	O
functional	NN	O	O
activity	NN	O	O
.	NN	O	O

A	NN	O	O
protein	NN	O	O
footprint	NN	O	O
was	NN	O	O
visible	NN	O	O
over	NN	O	O
this	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
c-myb	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
sequence	NN	O	I-DNA
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
unactivated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

An	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
with	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
from	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
an	NN	O	O
oligonucleotide	NN	O	O
of	NN	O	O
this	NN	O	O
binding	NN	O	O
site	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
new	NN	O	O
protein-DNA	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
CMAT	NN	O	B-protein
for	NN	O	O
c-myb	NN	O	B-protein
in	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
;	NN	O	O
this	NN	O	O
complex	NN	O	O
was	NN	O	O
not	NN	O	O
present	NN	O	O
in	NN	O	O
unactivated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Because	NN	O	O
the	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
showed	NN	O	O
some	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
with	NN	O	O
the	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
of	NN	O	I-DNA
activated	NN	O	I-DNA
T	NN	O	I-DNA
cells	NN	O	I-DNA
(	NN	O	I-DNA
NFAT	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
compared	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
binding	NN	O	O
complexes	NN	O	O
and	NN	O	O
the	NN	O	O
molecular	NN	O	O
masses	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
proteins	NN	O	O
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
induction	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
EMSA	NN	O	I-protein
binding	NN	O	I-protein
complex	NN	O	I-protein
appeared	NN	O	O
earlier	NN	O	O
than	NN	O	O
the	NN	O	O
CMAT	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

The	NN	O	O
NFAT	NN	O	B-protein
protein	NN	O	O
migrated	NN	O	O
more	NN	O	O
slowly	NN	O	O
in	NN	O	O
a	NN	O	O
sodium	NN	O	O
dodecyl	NN	O	O
sulfate-polyacrylamide	NN	O	O
gel	NN	O	O
than	NN	O	O
the	NN	O	O
CMAT	NN	O	B-protein
protein	NN	O	I-protein
did	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
an	NN	O	O
antibody	NN	O	O
against	NN	O	O
NFAT	NN	O	B-protein
did	NN	O	O
not	NN	O	O
cross-react	NN	O	O
with	NN	O	O
the	NN	O	O
CMAT	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
appearance	NN	O	O
of	NN	O	O
the	NN	O	O
CMAT	NN	O	B-protein
binding	NN	O	I-protein
complex	NN	O	I-protein
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
both	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
rapamycin	NN	O	O
.	NN	O	O

The	NN	O	O
CMAT	NN	O	B-protein
protein	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
novel	NN	O	O
inducible	NN	O	O
protein	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
c-myb	NN	O	B-DNA
expression	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

CNI-1493	NN	O	O
inhibits	NN	O	O
monocyte/macrophage	NN	O	B-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
by	NN	O	O
suppression	NN	O	O
of	NN	O	O
translation	NN	O	O
efficiency	NN	O	O
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
mediates	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
disease	NN	O	O
states	NN	O	O
including	NN	O	O
septic	NN	O	O
shock	NN	O	O
,	NN	O	O
acute	NN	O	O
and	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
,	NN	O	O
and	NN	O	O
cachexia	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
a	NN	O	O
multivalent	NN	O	O
guanylhydrazone	NN	O	O
(	NN	O	O
CNI-1493	NN	O	O
)	NN	O	O
developed	NN	O	O
as	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
macrophage	NN	O	O
activation	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
suppress	NN	O	O
TNF	NN	O	B-protein
production	NN	O	O
and	NN	O	O
protect	NN	O	O
against	NN	O	O
tissue	NN	O	O
inflammation	NN	O	O
and	NN	O	O
endotoxin	NN	O	O
lethality	NN	O	O
[	NN	O	O
Bianchi	NN	O	O
,	NN	O	O
M.	NN	O	O
,	NN	O	O
Ulrich	NN	O	O
,	NN	O	O
P.	NN	O	O
,	NN	O	O
Bloom	NN	O	O
,	NN	O	O
O.	NN	O	O
,	NN	O	O
Meistrell	NN	O	O
,	NN	O	O
M.	NN	O	O
,	NN	O	O
Zimmerman	NN	O	O
,	NN	O	O
G.A.	NN	O	O
,	NN	O	O
Schmidtmayerova	NN	O	O
,	NN	O	O
H.	NN	O	O
,	NN	O	O
Bukrinsky	NN	O	O
,	NN	O	O
M.	NN	O	O
,	NN	O	O
Donnelley	NN	O	O
,	NN	O	O
T.	NN	O	O
,	NN	O	O
Bucala	NN	O	O
,	NN	O	O
R.	NN	O	O
,	NN	O	O
Sherry	NN	O	O
,	NN	O	O
B.	NN	O	O
,	NN	O	O
Manogue	NN	O	O
,	NN	O	O
K.R.	NN	O	O
,	NN	O	O
Tortolani	NN	O	O
,	NN	O	O
A.J.	NN	O	O
,	NN	O	O
Cerami	NN	O	O
,	NN	O	O
A.	NN	O	O
&	NN	O	O
Tracey	NN	O	O
,	NN	O	O
K.J.	NN	O	O
(	NN	O	O
1995	NN	O	O
)	NN	O	O
Mol.Med.1	NN	O	O
,	NN	O	O
254-266	NN	O	O
,	NN	O	O
and	NN	O	O
Bianchi	NN	O	O
,	NN	O	O
M.	NN	O	O
,	NN	O	O
Bloom	NN	O	O
,	NN	O	O
O.	NN	O	O
,	NN	O	O
Raabe	NN	O	O
,	NN	O	O
T.	NN	O	O
,	NN	O	O
Cohen	NN	O	O
,	NN	O	O
P.	NN	O	O
S.	NN	O	O
,	NN	O	O
Chesney	NN	O	O
,	NN	O	O
J.	NN	O	O
,	NN	O	O
Sherry	NN	O	O
,	NN	O	O
B.	NN	O	O
,	NN	O	O
Schmidtmayerova	NN	O	O
,	NN	O	O
H.	NN	O	O
,	NN	O	O
Zhang	NN	O	O
,	NN	O	O
X.	NN	O	O
,	NN	O	O
Bukrinsky	NN	O	O
,	NN	O	O
M.	NN	O	O
,	NN	O	O
Ulrich	NN	O	O
,	NN	O	O
P.	NN	O	O
,	NN	O	O
Cerami	NN	O	O
,	NN	O	O
A.	NN	O	O
&	NN	O	O
Tracey	NN	O	O
,	NN	O	O
J.	NN	O	O
(	NN	O	O
1996	NN	O	O
)	NN	O	O
J.Exp.Med.	NN	O	O
,	NN	O	O
in	NN	O	O
press	NN	O	O
]	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
elucidated	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
CNI-1493	NN	O	O
inhibits	NN	O	O
macrophage	NN	O	O
TNF	NN	O	B-protein
synthesis	NN	O	O
and	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
it	NN	O	O
acts	NN	O	O
through	NN	O	O
suppression	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
translation	NN	O	O
efficiency	NN	O	O
.	NN	O	O

CNI-1493	NN	O	O
blocked	NN	O	O
neither	NN	O	O
the	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-induced	NN	O	O
increases	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
TNF	NN	O	B-RNA
mRNA	NN	O	I-RNA
nor	NN	O	O
the	NN	O	O
translocation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
activated	NN	O	O
by	NN	O	O
15	NN	O	O
min	NN	O	O
of	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
CNI-1493	NN	O	O
does	NN	O	O
not	NN	O	O
interfere	NN	O	O
with	NN	O	O
early	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-mediated	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
synthesis	NN	O	O
of	NN	O	O
the	NN	O	O
26-kDa	NN	O	B-protein
membrane	NN	O	I-protein
form	NN	O	I-protein
of	NN	O	O
TNF	NN	O	B-protein
was	NN	O	O
effectively	NN	O	O
blocked	NN	O	O
by	NN	O	O
CNI-1493	NN	O	O
.	NN	O	O

Further	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
translational	NN	O	O
suppression	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
is	NN	O	O
given	NN	O	O
by	NN	O	O
experiments	NN	O	O
using	NN	O	O
chloram-phenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
constructs	NN	O	I-DNA
containing	NN	O	O
elements	NN	O	O
of	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
TNF	NN	O	B-protein
translational	NN	O	O
regulation	NN	O	O
.	NN	O	O

Both	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
and	NN	O	I-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
untranslated	NN	O	I-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
gene	NN	O	I-DNA
were	NN	O	O
required	NN	O	O
to	NN	O	O
elicit	NN	O	O
maximal	NN	O	O
translational	NN	O	O
suppression	NN	O	O
by	NN	O	O
CNI-1493	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
target	NN	O	O
through	NN	O	O
which	NN	O	O
CNI-	NN	O	O
1493	NN	O	O
inhibits	NN	O	O
TNF	NN	O	B-protein
translation	NN	O	O
should	NN	O	O
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
macrophage	NN	O	O
activation	NN	O	O
and	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
inflammation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Soluble	NN	O	B-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptors	NN	O	I-protein
inhibit	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
and	NN	O	O
cytokine-induced	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
chronically	NN	O	O
infected	NN	O	B-cell_type
U1	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Recombinant	NN	O	B-protein
human	NN	O	I-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
binding	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
r-h	NN	O	B-protein
TBP-1	NN	O	I-protein
)	NN	O	O
and	NN	O	O
recombinant	NN	O	B-protein
human	NN	O	I-protein
soluble	NN	O	I-protein
dimeric	NN	O	I-protein
TNF	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
rhu	NN	O	B-protein
TNFR	NN	O	I-protein
:	NN	O	I-protein
Fc	NN	O	I-protein
)	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
relative	NN	O	O
contributions	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
to	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
cytokine-induced	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
replication	NN	O	O
in	NN	O	O
chronically	NN	O	O
infected	NN	O	O
cell	NN	O	O
lines	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
HIV-1-infected	NN	O	B-cell_type
promonocytic	NN	O	I-cell_type
U1	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
r-h-TBP-1	NN	O	B-protein
or	NN	O	O
rhu	NN	O	B-protein
TNFR	NN	O	I-protein
:	NN	O	I-protein
Fc	NN	O	I-protein
reduced	NN	O	O
PMA-induced	NN	O	O
HIV-1	NN	O	B-protein
p24	NN	O	I-protein
antigen	NN	O	I-protein
production	NN	O	O
in	NN	O	O
a	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
maximal	NN	O	O
inhibition	NN	O	O
of	NN	O	O
approximately	NN	O	O
90	NN	O	O
%	NN	O	O
.	NN	O	O

Maximal	NN	O	O
inhibition	NN	O	O
of	NN	O	O
p24	NN	O	B-protein
antigen	NN	O	I-protein
production	NN	O	O
in	NN	O	O
T-lymphocytic	NN	O	B-cell_line
ACH-2	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
47	NN	O	O
%	NN	O	O
with	NN	O	O
r-hTBP-1	NN	O	B-protein
and	NN	O	O
42	NN	O	O
%	NN	O	O
with	NN	O	O
rhu	NN	O	B-protein
TNFR	NN	O	I-protein
:	NN	O	I-protein
Fc	NN	O	I-protein
.	NN	O	O

r-hTBP-1	NN	O	B-protein
and	NN	O	O
rhu	NN	O	B-protein
TNFR	NN	O	I-protein
:	NN	O	I-protein
Fc	NN	O	I-protein
also	NN	O	O
decreased	NN	O	O
p24	NN	O	B-protein
antigen	NN	O	I-protein
synthesized	NN	O	O
by	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
response	NN	O	O
to	NN	O	O
other	NN	O	O
stimuli	NN	O	O
,	NN	O	O
including	NN	O	O
phytohemagglutinin	NN	O	B-protein
(	NN	O	I-protein
PHA	NN	O	I-protein
)	NN	O	I-protein
-induced	NN	O	I-protein
supernatant	NN	O	I-protein
,	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
interleukin-6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TNF	NN	O	B-protein
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
r-hTBP-1	NN	O	B-protein
to	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
during	NN	O	O
the	NN	O	O
last	NN	O	O
4	NN	O	O
h	NN	O	O
of	NN	O	O
a	NN	O	O
24	NN	O	O
h	NN	O	O
incubation	NN	O	O
with	NN	O	O
PMA	NN	O	O
still	NN	O	O
inhibited	NN	O	O
p24	NN	O	B-protein
antigen	NN	O	I-protein
production	NN	O	O
by	NN	O	O
15	NN	O	O
%	NN	O	O
.	NN	O	O

U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
stimulated	NN	O	O
with	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
M	NN	O	O
PMA	NN	O	O
released	NN	O	O
approximately	NN	O	O
1	NN	O	O
ng/ml	NN	O	O
endogenous	NN	O	B-protein
TBP-1	NN	O	I-protein
with	NN	O	O
an	NN	O	O
initial	NN	O	O
peak	NN	O	O
observed	NN	O	O
at	NN	O	O
1	NN	O	O
h	NN	O	O
and	NN	O	O
a	NN	O	O
second	NN	O	O
peak	NN	O	O
at	NN	O	O
24	NN	O	O
h	NN	O	O
after	NN	O	O
PMA	NN	O	O
stimulation	NN	O	O
.	NN	O	O

r-hTBP-1	NN	O	B-protein
also	NN	O	O
partially	NN	O	O
reversed	NN	O	O
inhibition	NN	O	O
of	NN	O	O
U1	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
caused	NN	O	O
by	NN	O	O
PMA	NN	O	O
.	NN	O	O

Both	NN	O	O
r-hTBP-1	NN	O	B-protein
and	NN	O	O
rhu	NN	O	B-protein
TNFR	NN	O	I-protein
:	NN	O	I-protein
Fc	NN	O	I-protein
blocked	NN	O	O
PMA	NN	O	O
induction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NK	NN	O	I-protein
)	NN	O	I-protein
-	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
association	NN	O	O
with	NN	O	O
decreases	NN	O	O
in	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
soluble	NN	O	O
TNF	NN	O	O
receptors	NN	O	O
can	NN	O	O
inhibit	NN	O	O
stimuli-induced	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
and	NN	O	O
NK-	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
chronically	NN	O	O
infected	NN	O	B-cell_type
U1	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
RNA	NN	O	B-DNA
polymerase	NN	O	I-DNA
III-dependent	NN	O	I-DNA
genes	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
tax	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus-encoded	NN	O	I-protein
tax	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
activator	NN	O	O
of	NN	O	O
many	NN	O	O
viral	NN	O	B-DNA
and	NN	O	I-DNA
cellular	NN	O	I-DNA
genes	NN	O	I-DNA
transcribed	NN	O	O
by	NN	O	O
RNA	NN	O	B-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
both	NN	O	O
chromatin	NN	O	B-DNA
and	NN	O	O
cell	NN	O	O
extracts	NN	O	O
derived	NN	O	O
from	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
virus	NN	O	I-cell_line
type	NN	O	I-cell_line
1-infected	NN	O	I-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
support	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
5S	NN	O	O
rRNA	NN	O	O
and	NN	O	O
tRNA	NN	O	O
gene	NN	O	O
transcription	NN	O	O
than	NN	O	O
chromatin	NN	O	B-DNA
or	NN	O	O
extracts	NN	O	O
from	NN	O	O
uninfected	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
viral	NN	O	B-protein
protein	NN	O	I-protein
Tax	NN	O	I-protein
was	NN	O	O
likely	NN	O	O
responsible	NN	O	O
for	NN	O	O
this	NN	O	O
higher	NN	O	O
level	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
,	NN	O	O
as	NN	O	O
purified	NN	O	B-protein
Tax	NN	O	I-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
stimulate	NN	O	O
both	NN	O	O
genes	NN	O	O
when	NN	O	O
added	NN	O	O
to	NN	O	O
the	NN	O	O
uninfected	NN	O	O
cell	NN	O	O
extract	NN	O	O
or	NN	O	O
in	NN	O	O
reconstituted	NN	O	O
systems	NN	O	O
.	NN	O	O

Both	NN	O	O
limiting-component	NN	O	O
transcription	NN	O	O
assays	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
assays	NN	O	O
identified	NN	O	O
the	NN	O	O
class	NN	O	B-protein
III	NN	O	I-protein
gene	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
TFIIIB	NN	O	B-protein
as	NN	O	O
the	NN	O	O
principle	NN	O	O
target	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
we	NN	O	O
find	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
increases	NN	O	O
the	NN	O	O
effective	NN	O	O
concentration	NN	O	O
of	NN	O	O
active	NN	O	B-protein
TFIIIB	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
stimulates	NN	O	O
RNA	NN	O	B-DNA
polymerase	NN	O	I-DNA
III-dependent	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
accelerating	NN	O	O
the	NN	O	O
rate	NN	O	O
and/or	NN	O	O
extent	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
initiation	NN	O	I-protein
complex	NN	O	I-protein
assembly	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
Retinoid	NN	O	B-protein
X	NN	O	I-protein
Receptor	NN	O	I-protein
alpha	NN	O	I-protein
is	NN	O	O
increased	NN	O	O
upon	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-Dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
VD	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
inducer	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
both	NN	O	O
normal	NN	O	B-cell_type
and	NN	O	I-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Its	NN	O	O
effects	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
its	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Efficient	NN	O	O
gene	NN	O	O
activation	NN	O	O
requires	NN	O	O
the	NN	O	O
heterodimerization	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
with	NN	O	O
Retinoid	NN	O	B-protein
X	NN	O	I-protein
Receptors	NN	O	I-protein
(	NN	O	O
RXR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
using	NN	O	O
specific	NN	O	B-protein
antibodies	NN	O	I-protein
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
RXR	NN	O	B-protein
alpha	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
blood	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
patients	NN	O	O
(	NN	O	O
AML	NN	O	O
)	NN	O	O
(	NN	O	O
10	NN	O	O
cases	NN	O	O
)	NN	O	O
and	NN	O	O
from	NN	O	O
myelomonocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
arrested	NN	O	O
at	NN	O	O
different	NN	O	O
stages	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
the	NN	O	O
RXR	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
increased	NN	O	O
during	NN	O	O
myelomonocytic	NN	O	O
differentiation	NN	O	O
,	NN	O	O
since	NN	O	O
the	NN	O	O
highest	NN	O	O
levels	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
AML	NN	O	O
samples	NN	O	O
and	NN	O	O
in	NN	O	O
myelomonocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
having	NN	O	O
the	NN	O	O
highest	NN	O	O
amounts	NN	O	O
of	NN	O	O
monocytic	NN	O	B-cell_type
precursors	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
fresh	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whatever	NN	O	O
their	NN	O	O
stage	NN	O	O
of	NN	O	O
differentiation	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
myelomonocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
respond	NN	O	O
to	NN	O	O
VD	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
RXR	NN	O	B-protein
alpha	NN	O	I-protein
levels	NN	O	O
.	NN	O	O

Combinations	NN	O	O
of	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
and	NN	O	O
VD	NN	O	O
,	NN	O	O
in	NN	O	O
some	NN	O	O
cases	NN	O	O
,	NN	O	O
increased	NN	O	O
this	NN	O	O
effect	NN	O	O
.	NN	O	O

This	NN	O	O
response	NN	O	O
suggests	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
RXR	NN	O	B-protein
alpha	NN	O	I-protein
in	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
upon	NN	O	O
VD	NN	O	O
treatment	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
p13MTCP1	NN	O	B-protein
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
mature	NN	O	O
T-cell	NN	O	O
proliferations	NN	O	O
with	NN	O	O
t	NN	O	O
(	NN	O	O
X	NN	O	O
;	NN	O	O
14	NN	O	O
)	NN	O	O
translocations	NN	O	O
.	NN	O	O

T-cell	NN	O	O
prolymphocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
T-PLL	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
rare	NN	O	O
form	NN	O	O
of	NN	O	O
mature	NN	O	O
T-cell	NN	O	O
leukemias	NN	O	O
,	NN	O	O
and	NN	O	O
ataxia	NN	O	O
telangiectasia	NN	O	O
clonal	NN	O	O
proliferation	NN	O	O
,	NN	O	O
a	NN	O	O
related	NN	O	O
condition	NN	O	O
occurring	NN	O	O
in	NN	O	O
patients	NN	O	O
suffering	NN	O	O
from	NN	O	O
ataxia	NN	O	O
telangiectasia	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
associated	NN	O	O
to	NN	O	O
translocations	NN	O	O
involving	NN	O	O
the	NN	O	O
14q32.1	NN	O	B-DNA
or	NN	O	O
Xq28	NN	O	B-DNA
regions	NN	O	I-DNA
,	NN	O	O
where	NN	O	O
are	NN	O	O
located	NN	O	O
the	NN	O	O
TCL1	NN	O	B-DNA
and	NN	O	I-DNA
MTCP1	NN	O	I-DNA
putative	NN	O	I-DNA
oncogenes	NN	O	I-DNA
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
MTCP1	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
X	NN	O	I-DNA
;	NN	O	I-DNA
14	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q28	NN	O	I-DNA
;	NN	O	I-DNA
q11	NN	O	I-DNA
)	NN	O	I-DNA
translocation	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
these	NN	O	O
T-cell	NN	O	O
proliferations	NN	O	O
.	NN	O	O

Alternative	NN	O	O
splicing	NN	O	O
generates	NN	O	O
type	NN	O	B-RNA
A	NN	O	I-RNA
and	NN	O	I-RNA
B	NN	O	I-RNA
transcripts	NN	O	I-RNA
that	NN	O	O
potentially	NN	O	O
encode	NN	O	O
two	NN	O	O
entirely	NN	O	O
distinct	NN	O	O
proteins	NN	O	O
;	NN	O	O
type	NN	O	O
A	NN	O	B-RNA
transcripts	NN	O	I-RNA
code	NN	O	O
for	NN	O	O
a	NN	O	O
small	NN	O	O
mitochondrial	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
p8MTCP1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
type	NN	O	B-RNA
B	NN	O	I-RNA
transcripts	NN	O	I-RNA
,	NN	O	O
containing	NN	O	O
an	NN	O	O
additional	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
,	NN	O	O
may	NN	O	O
code	NN	O	O
for	NN	O	O
107	NN	O	B-protein
amino-acid	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
p13MTCP1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
recently	NN	O	O
cloned	NN	O	O
TCL1	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
also	NN	O	O
involved	NN	O	O
in	NN	O	O
translocations	NN	O	O
and	NN	O	O
inversions	NN	O	O
associated	NN	O	O
with	NN	O	O
T-cell	NN	O	O
proliferations	NN	O	O
,	NN	O	O
codes	NN	O	O
for	NN	O	O
a	NN	O	O
14-kD	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
displays	NN	O	O
significant	NN	O	O
homology	NN	O	O
with	NN	O	O
p13MTCP1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
generated	NN	O	O
rabbit	NN	O	O
antisera	NN	O	O
against	NN	O	O
this	NN	O	O
putative	NN	O	O
p13MTCP1	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
screened	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
p13MTCP1	NN	O	B-protein
normal	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
and	NN	O	O
33	NN	O	O
cases	NN	O	O
of	NN	O	O
immature	NN	O	B-cell_type
and	NN	O	I-cell_type
mature	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
T-cell	NN	O	I-cell_type
proliferations	NN	O	I-cell_type
using	NN	O	O
a	NN	O	O
sensitive	NN	O	O
Western	NN	O	O
blot	NN	O	O
assay	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
investigated	NN	O	O
the	NN	O	O
MTCP1	NN	O	B-DNA
locus	NN	O	I-DNA
configuration	NN	O	O
by	NN	O	O
Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
p13MTCP1	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
three	NN	O	O
T-cell	NN	O	O
proliferations	NN	O	O
with	NN	O	O
MTCP1	NN	O	B-DNA
rearrangements	NN	O	I-DNA
because	NN	O	O
of	NN	O	O
t	NN	O	O
(	NN	O	O
X	NN	O	O
;	NN	O	O
14	NN	O	O
)	NN	O	O
translocations	NN	O	O
,	NN	O	O
but	NN	O	O
neither	NN	O	O
in	NN	O	O
normal	NN	O	O
resting	NN	O	O
and	NN	O	O
activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
nor	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
T-cell	NN	O	B-cell_type
leukemias	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
p13MTCP1	NN	O	B-protein
and	NN	O	O
p14TCL1	NN	O	B-protein
form	NN	O	O
a	NN	O	O
new	NN	O	O
protein	NN	O	B-protein
family	NN	O	I-protein
that	NN	O	O
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
T-PLL	NN	O	O
and	NN	O	O
related	NN	O	O
conditions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Preassociation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
with	NN	O	O
STAT2	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
in	NN	O	O
separate	NN	O	O
signalling	NN	O	B-protein
complexes	NN	O	I-protein
prior	NN	O	O
to	NN	O	O
cytokine	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

A	NN	O	O
variety	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
act	NN	O	O
through	NN	O	O
an	NN	O	O
induction	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
latent	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
called	NN	O	O
STAT	NN	O	B-protein
(	NN	O	I-protein
signal	NN	O	I-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Ligand-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
STATs	NN	O	B-protein
promotes	NN	O	O
their	NN	O	O
homodimer	NN	O	O
and	NN	O	O
heterodimer	NN	O	O
formation	NN	O	O
and	NN	O	O
subsequent	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
that	NN	O	O
STAT	NN	O	B-protein
protein	NN	O	I-protein
heterocomplexes	NN	O	I-protein
exist	NN	O	O
prior	NN	O	O
to	NN	O	O
cytokine	NN	O	B-protein
treatment	NN	O	O
.	NN	O	O

When	NN	O	O
unstimulated	NN	O	B-cell_line
HeLa	NN	O	I-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
ruptured	NN	O	O
in	NN	O	O
hypotonic	NN	O	O
buffer	NN	O	O
without	NN	O	O
salt	NN	O	O
or	NN	O	O
detergent	NN	O	O
,	NN	O	O
immunoadsorption	NN	O	O
of	NN	O	O
either	NN	O	O
STAT1	NN	O	B-protein
or	NN	O	O
STAT2	NN	O	B-protein
from	NN	O	O
the	NN	O	O
resulting	NN	O	O
cytosol	NN	O	O
yields	NN	O	O
coimmunoadsorption	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
STAT	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
STAT1	NN	O	B-protein
-STAT3	NN	O	B-protein
heterocomplexes	NN	O	O
are	NN	O	O
coimmunoadsorbed	NN	O	O
from	NN	O	O
hypotonic	NN	O	O
cytosol	NN	O	O
.	NN	O	O

STAT1	NN	O	B-protein
and	NN	O	O
STAT2	NN	O	B-protein
or	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
translated	NN	O	O
in	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
spontaneously	NN	O	O
form	NN	O	O
heterocomplexes	NN	O	O
when	NN	O	O
the	NN	O	O
translation	NN	O	O
lysates	NN	O	O
are	NN	O	O
mixed	NN	O	O
at	NN	O	O
0	NN	O	O
degrees	NN	O	O
C	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
interferon-alpha	NN	O	B-protein
/beta-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
increases	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
a	NN	O	O
preexisting	NN	O	O
,	NN	O	O
latent	NN	O	O
,	NN	O	O
STAT1	NN	O	B-protein
-	NN	O	O
STAT2	NN	O	B-protein
signaling	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Newly	NN	O	O
translated	NN	O	O
STAT1	NN	O	B-protein
binds	NN	O	O
in	NN	O	O
equilibrium	NN	O	O
fashion	NN	O	O
to	NN	O	O
STAT2	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
STAT2	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
exist	NN	O	O
in	NN	O	O
separate	NN	O	O
heterocomplexes	NN	O	B-protein
with	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
STAT1	NN	O	B-protein
contains	NN	O	O
a	NN	O	O
common	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
other	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Interferon-gamma	NN	O	B-protein
modulates	NN	O	O
the	NN	O	O
lipopolysaccharide-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
level	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
modulates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
cytokines	NN	O	B-protein
by	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

In	NN	O	O
view	NN	O	O
of	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
different	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
activated	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Priming	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
with	NN	O	O
IFN-gamma	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
down	NN	O	O
regulation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
and	NN	O	I-RNA
c-jun	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
stimulation	NN	O	O
with	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
LPS	NN	O	O
alone	NN	O	O
.	NN	O	O

Not	NN	O	O
only	NN	O	O
was	NN	O	O
this	NN	O	O
effect	NN	O	O
observed	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
,	NN	O	O
but	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
DNA	NN	O	O
binding	NN	O	O
capacity	NN	O	O
was	NN	O	O
affected	NN	O	O
as	NN	O	O
well	NN	O	O
,	NN	O	O
A	NN	O	O
strong	NN	O	O
reduction	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
LPS-induced	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

LPS-stimulated	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
showed	NN	O	O
an	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
p105	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
the	NN	O	O
precursor	NN	O	O
of	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
while	NN	O	O
no	NN	O	O
effect	NN	O	O
was	NN	O	O
noticed	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
p65	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
IFN-gamma	NN	O	B-protein
priming	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
p105	NN	O	B-RNA
transcripts	NN	O	I-RNA
but	NN	O	O
enhanced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
p65	NN	O	B-RNA
mRNA	NN	O	I-RNA
(	NN	O	O
two-fold	NN	O	O
)	NN	O	O
.	NN	O	O

Priming	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
followed	NN	O	O
by	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
further	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
p65	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

This	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
half-life	NN	O	O
of	NN	O	O
p65	NN	O	B-RNA
mRNA	NN	O	I-RNA
(	NN	O	O
75	NN	O	O
vs	NN	O	O
150	NN	O	O
minutes	NN	O	O
)	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSAs	NN	O	O
)	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
unstimulated	NN	O	O
monocytes	NN	O	O
predominantly	NN	O	O
expressed	NN	O	O
p50	NN	O	B-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Stimulation	NN	O	O
with	NN	O	O
LPS	NN	O	O
or	NN	O	O
IFN-gamma	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
subunits	NN	O	I-protein
,	NN	O	O
while	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
plus	NN	O	O
LPS	NN	O	O
caused	NN	O	O
a	NN	O	O
further	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

With	NN	O	O
Western	NN	O	O
blotting	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
shown	NN	O	O
that	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
monocytes	NN	O	B-cell_type
contained	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
combined	NN	O	O
stimulation	NN	O	O
did	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
enhanced	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
protein	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
on	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
were	NN	O	O
specific	NN	O	O
,	NN	O	O
since	NN	O	O
no	NN	O	O
change	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-IL-6	NN	O	B-protein
or	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
the	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
may	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
modulatory	NN	O	O
effects	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
on	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
activated	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

HIV-1	NN	O	B-protein
tat	NN	O	I-protein
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
hematopoietic	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
downregulates	NN	O	O
MIP-1	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

MIP-1	NN	O	B-protein
alpha	NN	O	I-protein
is	NN	O	O
a	NN	O	O
secreted	NN	O	O
chemokine	NN	O	B-protein
which	NN	O	O
can	NN	O	O
inhibit	NN	O	O
hematopoietic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
modulate	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
also	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
CD8+	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
MIP-1	NN	O	B-protein
alpha	NN	O	I-protein
is	NN	O	O
induced	NN	O	O
during	NN	O	O
cellular	NN	O	O
activation	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

It	NN	O	O
is	NN	O	O
also	NN	O	O
expressed	NN	O	O
in	NN	O	O
transformed	NN	O	B-cell_line
B-cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
identified	NN	O	O
a	NN	O	O
new	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
family	NN	O	I-protein
(	NN	O	O
the	NN	O	O
MNP	NN	O	B-protein
family	NN	O	I-protein
)	NN	O	O
whose	NN	O	O
expression	NN	O	O
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
MIP-1	NN	O	B-protein
alpha	NN	O	I-protein
transcription	NN	O	O
during	NN	O	O
cellular	NN	O	O
activation	NN	O	O
and	NN	O	O
in	NN	O	O
transformed	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Monocytes	NN	O	O
and	NN	O	O
transformed	NN	O	B-cell_line
B-cells	NN	O	I-cell_line
normally	NN	O	O
express	NN	O	O
MNP-1	NN	O	B-protein
strongly	NN	O	O
and	NN	O	O
MNP-2	NN	O	B-protein
weakly	NN	O	O
,	NN	O	O
while	NN	O	O
T-cells	NN	O	O
strongly	NN	O	O
express	NN	O	O
only	NN	O	O
MNP-2	NN	O	B-protein
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
reported	NN	O	O
that	NN	O	O
HIV-1	NN	O	B-protein
tat	NN	O	I-protein
downregulates	NN	O	O
MIP-1	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
show	NN	O	O
induction	NN	O	O
of	NN	O	O
MNP-1	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
expressing	NN	O	O
HIV-1	NN	O	B-protein
tat	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
neither	NN	O	O
HTLV-1	NN	O	B-protein
tax	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
nor	NN	O	O
EBV	NN	O	O
in	NN	O	O
B-cells	NN	O	B-cell_type
had	NN	O	O
any	NN	O	O
effect	NN	O	O
on	NN	O	O
MNP-1	NN	O	B-protein
or	NN	O	O
MNP-2	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
showing	NN	O	O
that	NN	O	O
the	NN	O	O
effect	NN	O	O
is	NN	O	O
specific	NN	O	O
for	NN	O	O
HIV-1	NN	O	B-protein
tat	NN	O	I-protein
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
HIV-1	NN	O	B-protein
tat	NN	O	I-protein
may	NN	O	O
inhibit	NN	O	O
MIP-1	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
inducing	NN	O	O
MNP-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
T-cells	NN	O	B-cell_type
,	NN	O	O
probably	NN	O	O
by	NN	O	O
either	NN	O	O
competing	NN	O	O
with	NN	O	O
MNP-2	NN	O	B-protein
for	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
MIP-1	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
or	NN	O	O
by	NN	O	O
sequestering	NN	O	O
it	NN	O	O
into	NN	O	O
inactive	NN	O	O
forms	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differentiation	NN	O	O
of	NN	O	O
T-helper	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
selective	NN	O	O
regulation	NN	O	O
by	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
STAT	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interleukin-12	NN	O	B-protein
(	NN	O	O
IL-12	NN	O	B-protein
)	NN	O	O
control	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
T-helper	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
summarize	NN	O	O
studies	NN	O	O
which	NN	O	O
investigate	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
selectively	NN	O	O
reprogramme	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T-lymphocytes	NN	O	B-cell_type
.	NN	O	O

Cytokine	NN	O	B-protein
stimulation	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
specific	NN	O	O
tyrosine	NN	O	O
residues	NN	O	O
within	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
corresponding	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

These	NN	O	O
phosphotyrosines	NN	O	O
serve	NN	O	O
as	NN	O	O
docking	NN	O	B-protein
sites	NN	O	I-protein
for	NN	O	O
latent	NN	O	O
,	NN	O	O
cytoplasmic	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
known	NN	O	O
as	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
Stat	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Receptor/Stat	NN	O	O
interaction	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
src	NN	O	B-protein
homology	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	I-protein
SH2	NN	O	I-protein
)	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
corresponding	NN	O	O
Stat	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
Stat	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
strongly	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
phosphotyrosine	NN	O	O
residue	NN	O	O
,	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
a	NN	O	O
specific	NN	O	O
Stat	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
dictated	NN	O	O
by	NN	O	O
amino	NN	O	O
acid	NN	O	O
residues	NN	O	O
C-terminal	NN	O	O
to	NN	O	O
the	NN	O	O
phosphotyrosine	NN	O	O
.	NN	O	O

Specific	NN	O	O
docking	NN	O	B-protein
sites	NN	O	I-protein
within	NN	O	O
individual	NN	O	O
cytokine	NN	O	B-protein
receptors	NN	O	I-protein
have	NN	O	O
been	NN	O	O
identified	NN	O	O
for	NN	O	O
almost	NN	O	O
all	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
direct	NN	O	O
coupling	NN	O	O
between	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
helps	NN	O	O
to	NN	O	O
explain	NN	O	O
how	NN	O	O
different	NN	O	O
cytokines	NN	O	B-protein
elicit	NN	O	O
distinct	NN	O	O
patterns	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Role	NN	O	O
of	NN	O	O
EGR1	NN	O	B-protein
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
stimulus-dependent	NN	O	O
CD44	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
B	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

The	NN	O	O
immediate-early	NN	O	B-DNA
gene	NN	O	I-DNA
egr-1	NN	O	B-DNA
encodes	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
EGR1	NN	O	B-protein
)	NN	O	O
that	NN	O	O
links	NN	O	O
B-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
BCR	NN	O	B-protein
)	NN	O	O
signals	NN	O	O
to	NN	O	O
downstream	NN	O	O
activation	NN	O	O
events	NN	O	O
through	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
previously	NN	O	O
unidentified	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
we	NN	O	O
identify	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
lymphocyte	NN	O	B-protein
homing	NN	O	I-protein
and	NN	O	I-protein
migration	NN	O	I-protein
protein	NN	O	I-protein
CD44	NN	O	B-protein
as	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
EGR1	NN	O	B-protein
regulation	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

BCR	NN	O	B-protein
-induced	NN	O	O
increases	NN	O	O
in	NN	O	O
CD44	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
transcription	NN	O	O
levels	NN	O	O
are	NN	O	O
shown	NN	O	O
to	NN	O	O
occur	NN	O	O
in	NN	O	O
EGR1-expressing	NN	O	B-cell_line
but	NN	O	I-cell_line
not	NN	O	I-cell_line
in	NN	O	I-cell_line
nonexpressing	NN	O	I-cell_line
subclones	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
B-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
WEHI-231	NN	O	B-cell_line
.	NN	O	O

Kinetics	NN	O	O
of	NN	O	O
egr-1	NN	O	B-DNA
transcription	NN	O	O
and	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
nuclear	NN	O	O
EGR1	NN	O	B-protein
protein	NN	O	I-protein
precede	NN	O	O
CD44	NN	O	B-protein
induction	NN	O	O
and	NN	O	O
occur	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
after	NN	O	O
stimulation	NN	O	O
in	NN	O	O
the	NN	O	O
EGR1-expressing	NN	O	B-cell_line
subclone	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
single	NN	O	O
EGR1	NN	O	B-protein
binding	NN	O	O
motif	NN	O	O
is	NN	O	O
demonstrated	NN	O	O
at	NN	O	O
bp	NN	O	O
-301	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD44	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
a	NN	O	O
CD44	NN	O	B-DNA
promoter-chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
with	NN	O	O
an	NN	O	O
egr-1	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
6.5-	NN	O	O
to	NN	O	O
8.5-fold	NN	O	O
induction	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
relative	NN	O	O
to	NN	O	O
an	NN	O	O
empty	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
EGR1	NN	O	B-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
necessary	NN	O	O
for	NN	O	O
stimulus-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
CD44	NN	O	B-protein
promoter-chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
in	NN	O	O
nontransformed	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
transactivation	NN	O	O
by	NN	O	O
an	NN	O	O
EGR1	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
B-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
studies	NN	O	O
identify	NN	O	O
EGR1	NN	O	B-protein
as	NN	O	O
an	NN	O	O
intermediary	NN	O	O
linking	NN	O	O
BCR	NN	O	B-protein
-derived	NN	O	O
signals	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
CD44	NN	O	B-protein
.	NN	O	O

The	NN	O	O
relevance	NN	O	O
of	NN	O	O
these	NN	O	O
molecular	NN	O	O
events	NN	O	O
to	NN	O	O
BCR	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
and	NN	O	O
antigen-stimulated	NN	O	O
B-cell-mediated	NN	O	O
immune	NN	O	O
responses	NN	O	O
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
in	NN	O	O
oxidant-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

In	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
its	NN	O	O
subclone	NN	O	O
Wurzburg	NN	O	B-cell_line
cells	NN	O	I-cell_line
oxidant	NN	O	O
challenge	NN	O	O
elevated	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
by	NN	O	O
mobilizing	NN	O	O
Ca2+	NN	O	O
from	NN	O	O
intracellular	NN	O	O
stores	NN	O	O
.	NN	O	O

In	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
this	NN	O	O
effect	NN	O	O
was	NN	O	O
rapid	NN	O	O
and	NN	O	O
transient	NN	O	O
,	NN	O	O
but	NN	O	O
in	NN	O	O
Wurzburg	NN	O	B-cell_line
cells	NN	O	I-cell_line
the	NN	O	O
response	NN	O	O
was	NN	O	O
slow	NN	O	O
and	NN	O	O
sustained	NN	O	O
.	NN	O	O

H2O2-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
Wurzburg	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
not	NN	O	O
influenced	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
extracellular	NN	O	O
EGTA	NN	O	O
but	NN	O	O
was	NN	O	O
totally	NN	O	O
inhibited	NN	O	O
in	NN	O	O
cells	NN	O	O
that	NN	O	O
were	NN	O	O
loaded	NN	O	O
with	NN	O	O
esterified	NN	O	O
EGTA	NN	O	O
.	NN	O	O

In	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
are	NN	O	O
not	NN	O	O
sensitive	NN	O	O
to	NN	O	O
H2O2-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
H2O2	NN	O	O
potentiated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
sustained	NN	O	O
high	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
following	NN	O	O
thapsigargin	NN	O	O
treatment	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
regulatory	NN	O	O
effect	NN	O	O
of	NN	O	O
alpha-lipoate	NN	O	O
and	NN	O	O
N-acetylcysteine	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
due	NN	O	O
to	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
stabilize	NN	O	O
elevation	NN	O	O
of	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
following	NN	O	O
oxidant	NN	O	O
challenge	NN	O	O
.	NN	O	O

Results	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
indicate	NN	O	O
that	NN	O	O
a	NN	O	O
sustained	NN	O	O
elevated	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
is	NN	O	O
a	NN	O	O
significant	NN	O	O
factor	NN	O	O
in	NN	O	O
oxidant-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-1alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
I	NN	O	I-protein
and	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
Tax	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
virus	NN	O	I-cell_line
type	NN	O	I-cell_line
I	NN	O	I-cell_line
(	NN	O	I-cell_line
HTLV-I	NN	O	I-cell_line
)	NN	O	I-cell_line
-infected	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
constitutively	NN	O	O
produce	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
interleukin-1alpha	NN	O	B-protein
(	NN	O	O
IL-1alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

To	NN	O	O
analyze	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-1alpha	NN	O	B-protein
in	NN	O	O
HTLV-I-infected	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
studied	NN	O	O
regulatory	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
IL-1alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1alpha	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

IL-1alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
drive	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
HTLV-I-positive	NN	O	B-cell_line
MT-2	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
constitutively	NN	O	O
produce	NN	O	O
IL-1alpha	NN	O	B-protein
.	NN	O	O

In	NN	O	O
a	NN	O	O
cotransfection	NN	O	O
assay	NN	O	O
,	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
both	NN	O	O
HTLV-I	NN	O	O
and	NN	O	O
HTLV-II	NN	O	O
specifically	NN	O	O
activated	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
IL-1alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
an	NN	O	O
uninfected	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
mutant	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
deficient	NN	O	O
in	NN	O	O
transactivation	NN	O	O
of	NN	O	O
genes	NN	O	O
by	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappaB	NN	O	I-protein
pathway	NN	O	O
was	NN	O	O
unable	NN	O	O
to	NN	O	O
induce	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
IL-1alpha	NN	O	B-DNA
promoter-CAT	NN	O	I-DNA
constructs	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
was	NN	O	O
rescued	NN	O	O
by	NN	O	O
exogenous	NN	O	O
provision	NN	O	O
of	NN	O	O
p65/p50	NN	O	B-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
two	NN	O	O
IL-1alpha	NN	O	B-DNA
kappaB-like	NN	O	I-DNA
sites	NN	O	I-DNA
(	NN	O	O
positions	NN	O	O
-1	NN	O	B-DNA
,	NN	O	I-DNA
065	NN	O	I-DNA
to	NN	O	I-DNA
-1	NN	O	I-DNA
,	NN	O	I-DNA
056	NN	O	I-DNA
and	NN	O	O
+646	NN	O	B-DNA
to	NN	O	I-DNA
+655	NN	O	I-DNA
)	NN	O	O
specifically	NN	O	O
formed	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
NF-kappaB	NN	O	B-protein
-containing	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
from	NN	O	O
MT-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
bound	NN	O	O
with	NN	O	O
higher	NN	O	O
affinity	NN	O	O
to	NN	O	O
the	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
than	NN	O	O
to	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
either	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
or	NN	O	I-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
sites	NN	O	I-DNA
reduced	NN	O	O
IL-1alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
MT-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
exogenous	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
Tax	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
a	NN	O	O
general	NN	O	O
role	NN	O	O
for	NN	O	O
Tax	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
IL-1alpha	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
by	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
IL-1alpha	NN	O	B-protein
by	NN	O	O
HTLV-I	NN	O	O
productively	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
hypercalcemia	NN	O	O
,	NN	O	O
osteolytic	NN	O	O
bone	NN	O	O
lesions	NN	O	O
,	NN	O	O
neutrophilia	NN	O	O
,	NN	O	O
elevation	NN	O	O
of	NN	O	O
C-reactive	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
fever	NN	O	O
frequently	NN	O	O
seen	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
HTLV-I-induced	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia/lymphoma	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
myeloid-cell-specific	NN	O	B-DNA
c-fes	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
novel	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
protein	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
c-fps/fes	NN	O	B-DNA
(	NN	O	I-DNA
c-fes	NN	O	I-DNA
)	NN	O	I-DNA
proto-oncogene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
development	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
neutrophils	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

mRNA	NN	O	O
for	NN	O	O
c-fes	NN	O	O
has	NN	O	O
been	NN	O	O
detected	NN	O	O
exclusively	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
vascular	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
adult	NN	O	O
mammals	NN	O	O
.	NN	O	O

Although	NN	O	O
a	NN	O	O
13-kilobase-pair	NN	O	B-DNA
(	NN	O	I-DNA
kb	NN	O	I-DNA
)	NN	O	I-DNA
human	NN	O	I-DNA
c-fes	NN	O	I-DNA
transgene	NN	O	I-DNA
exhibits	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
expression	NN	O	O
in	NN	O	O
mice	NN	O	O
,	NN	O	O
the	NN	O	O
sequences	NN	O	O
that	NN	O	O
confer	NN	O	O
myeloid-cell-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
c-fes	NN	O	I-DNA
gene	NN	O	I-DNA
have	NN	O	O
not	NN	O	O
been	NN	O	O
defined	NN	O	O
.	NN	O	O

Transient-transfection	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
plasmids	NN	O	B-DNA
containing	NN	O	O
446	NN	O	O
bp	NN	O	O
of	NN	O	O
c-fes	NN	O	B-DNA
5'-flanking	NN	O	I-DNA
sequences	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
a	NN	O	O
luciferase	NN	O	O
reporter	NN	O	O
gene	NN	O	O
were	NN	O	O
active	NN	O	O
exclusively	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

No	NN	O	O
other	NN	O	O
DNA	NN	O	O
element	NN	O	O
within	NN	O	O
the	NN	O	O
13-kb	NN	O	B-DNA
human	NN	O	I-DNA
c-fes	NN	O	I-DNA
locus	NN	O	I-DNA
contained	NN	O	O
positive	NN	O	B-DNA
cis-acting	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
a	NN	O	O
weakly	NN	O	O
active	NN	O	O
region	NN	O	O
within	NN	O	O
the	NN	O	O
3'-flanking	NN	O	B-DNA
sequences	NN	O	I-DNA
.	NN	O	O

DNase	NN	O	O
I	NN	O	O
footprinting	NN	O	O
assays	NN	O	O
revealed	NN	O	O
four	NN	O	O
distinct	NN	O	O
sites	NN	O	O
that	NN	O	O
bind	NN	O	O
myeloid	NN	O	B-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
-408	NN	O	B-DNA
to	NN	O	I-DNA
-386	NN	O	I-DNA
,	NN	O	O
-293	NN	O	B-DNA
to	NN	O	I-DNA
-254	NN	O	I-DNA
,	NN	O	O
-76	NN	O	B-DNA
to	NN	O	I-DNA
-65	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
-34	NN	O	B-DNA
to	NN	O	I-DNA
+3	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
first	NN	O	O
two	NN	O	O
footprints	NN	O	O
resided	NN	O	O
in	NN	O	O
sequences	NN	O	O
that	NN	O	O
were	NN	O	O
largely	NN	O	O
dispensable	NN	O	O
for	NN	O	O
transient	NN	O	O
activity	NN	O	O
.	NN	O	O

Plasmids	NN	O	O
containing	NN	O	O
151	NN	O	B-DNA
bp	NN	O	I-DNA
of	NN	O	O
5'-flanking	NN	O	B-DNA
sequences	NN	O	I-DNA
confer	NN	O	O
myeloid-cell-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analyses	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
151-bp	NN	O	B-DNA
region	NN	O	I-DNA
contains	NN	O	O
nuclear	NN	O	B-DNA
protein	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
and/or	NN	O	O
Elf-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

This	NN	O	O
unidentified	NN	O	O
factor	NN	O	O
binds	NN	O	O
immediately	NN	O	O
3	NN	O	O
'	NN	O	O
of	NN	O	O
the	NN	O	O
PU.1/Elf-1	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
specific	NN	O	O
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
PU.1	NN	O	B-protein
/Elf-1	NN	O	B-protein
site	NN	O	O
or	NN	O	O
the	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
FP4-3	NN	O	B-DNA
'	NN	O	I-DNA
)	NN	O	O
within	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
c-fes	NN	O	B-DNA
promoter	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
substantially	NN	O	O
reduced	NN	O	O
activity	NN	O	O
in	NN	O	O
transient	NN	O	O
transfections	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
transient-cotransfection	NN	O	O
assay	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
PU.1	NN	O	B-protein
(	NN	O	O
and	NN	O	O
not	NN	O	O
Elf-1	NN	O	B-protein
)	NN	O	O
can	NN	O	O
transactivate	NN	O	O
the	NN	O	O
c-fes	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
nonmyeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
c-fes	NN	O	I-DNA
gene	NN	O	I-DNA
contains	NN	O	O
a	NN	O	O
strong	NN	O	O
myeloid-cell-specific	NN	O	B-DNA
promoter	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
novel	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

IL-10	NN	O	B-protein
inhibits	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B/Rel	NN	O	I-protein
nuclear	NN	O	O
activity	NN	O	O
in	NN	O	O
CD3-stimulated	NN	O	B-cell_line
human	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

IL-10	NN	O	B-protein
markedly	NN	O	O
reduces	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
(	NN	O	B-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B/Rel	NN	O	I-protein
nuclear	NN	O	O
activity	NN	O	O
induced	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
the	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
OKT3	NN	O	B-protein
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
is	NN	O	O
exerted	NN	O	O
specifically	NN	O	O
on	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
mAb	NN	O	B-protein
OKT3	NN	O	I-protein
,	NN	O	O
and	NN	O	O
not	NN	O	O
that	NN	O	O
produced	NN	O	O
by	NN	O	O
PMA	NN	O	O
.	NN	O	O

As	NN	O	O
judged	NN	O	O
by	NN	O	O
supershifting	NN	O	O
the	NN	O	O
DNA-protein	NN	O	B-protein
complexes	NN	O	I-protein
with	NN	O	O
Abs	NN	O	B-protein
recognizing	NN	O	O
specific	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
protein	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
the	NN	O	O
p50/p65	NN	O	B-protein
(	NN	O	O
Rel	NN	O	B-protein
A	NN	O	I-protein
)	NN	O	O
heterodimeric	NN	O	O
form	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
primarily	NN	O	O
affected	NN	O	O
.	NN	O	O

The	NN	O	O
maximal	NN	O	O
effect	NN	O	O
is	NN	O	O
observed	NN	O	O
at	NN	O	O
the	NN	O	O
IL-10	NN	O	B-protein
concentration	NN	O	O
of	NN	O	O
20	NN	O	O
U/ml	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
inhibitory	NN	O	O
activity	NN	O	O
is	NN	O	O
exerted	NN	O	O
on	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
monocytes	NN	O	B-cell_type
pretreated	NN	O	O
with	NN	O	O
IL-10	NN	O	B-protein
are	NN	O	O
able	NN	O	O
so	NN	O	O
inhibit	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
nuclear	NN	O	O
activity	NN	O	O
in	NN	O	O
purified	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
OKT3	NN	O	B-protein
.	NN	O	O

Soluble	NN	O	O
factors	NN	O	O
do	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
inhibition	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
CD80	NN	O	B-protein
Ag	NN	O	I-protein
,	NN	O	O
found	NN	O	O
on	NN	O	O
monocytes	NN	O	B-cell_type
obtained	NN	O	O
from	NN	O	O
PBMC	NN	O	B-cell_type
incubated	NN	O	O
with	NN	O	O
OKT3	NN	O	B-protein
,	NN	O	O
is	NN	O	O
not	NN	O	O
detected	NN	O	O
after	NN	O	O
addition	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
CLB-CD28/1	NN	O	B-protein
restores	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
nuclear	NN	O	O
activity	NN	O	O
in	NN	O	O
IL-10-inhibited	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
inhibition	NN	O	O
might	NN	O	O
be	NN	O	O
ascribed	NN	O	O
to	NN	O	O
a	NN	O	O
lack	NN	O	O
of	NN	O	O
cooperation	NN	O	O
between	NN	O	O
accessory	NN	O	O
cells	NN	O	O
and	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
resulting	NN	O	O
from	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
a	NN	O	O
costimulatory	NN	O	O
molecule	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
CD80	NN	O	B-protein
,	NN	O	O
produced	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
on	NN	O	O
activated	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
can	NN	O	O
inhibit	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
nuclear	NN	O	O
activity	NN	O	O
in	NN	O	O
CD3-stimulated	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Since	NN	O	O
inappropriate	NN	O	O
activation	NN	O	O
of	NN	O	O
kappa	NN	O	B-DNA
B-driven	NN	O	I-DNA
genes	NN	O	I-DNA
has	NN	O	O
a	NN	O	O
physiopathologic	NN	O	O
role	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
diseases	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
HIV	NN	O	O
infection	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
support	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
using	NN	O	O
this	NN	O	O
cytokine	NN	O	O
to	NN	O	O
suppress	NN	O	O
an	NN	O	O
undesirable	NN	O	O
activation	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
A-myb	NN	O	B-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
c-myb	NN	O	B-DNA
and	NN	O	O
B-myb	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
,	NN	O	O
sIg+	NN	O	O
B-acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemias	NN	O	O
.	NN	O	O

The	NN	O	O
A-myb	NN	O	B-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
is	NN	O	O
related	NN	O	O
both	NN	O	O
functionally	NN	O	O
and	NN	O	O
structurally	NN	O	O
to	NN	O	O
the	NN	O	O
v-myb	NN	O	B-DNA
oncogene	NN	O	I-DNA
.	NN	O	O

Following	NN	O	O
our	NN	O	O
observations	NN	O	O
that	NN	O	O
A-myb	NN	O	B-DNA
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
restricted	NN	O	O
subset	NN	O	O
of	NN	O	O
normal	NN	O	O
mature	NN	O	B-cell_type
human	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
with	NN	O	O
the	NN	O	O
phenotype	NN	O	O
CD38+	NN	O	O
,	NN	O	O
CD39-	NN	O	O
,	NN	O	O
slgM-	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
now	NN	O	O
investigated	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
A-myb	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
neoplastic	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
representating	NN	O	O
the	NN	O	O
whole	NN	O	O
spectrum	NN	O	O
of	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
compared	NN	O	O
it	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
c-myb	NN	O	B-DNA
and	NN	O	O
B-myb	NN	O	B-DNA
.	NN	O	O

In	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
32	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
A-myb	NN	O	B-DNA
was	NN	O	O
very	NN	O	O
strongly	NN	O	O
expressed	NN	O	O
in	NN	O	O
most	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
BL	NN	O	O
)	NN	O	O
cell	NN	O	O
lines	NN	O	O
,	NN	O	O
but	NN	O	O
weak	NN	O	O
or	NN	O	O
negative	NN	O	O
in	NN	O	O
2	NN	O	O
pre-B	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ALL	NN	O	O
)	NN	O	O
,	NN	O	O
4	NN	O	O
non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
NHL	NN	O	O
)	NN	O	O
,	NN	O	O
6	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus-immortalized	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
6	NN	O	O
myeloma	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Protein	NN	O	O
expression	NN	O	O
paralleled	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
RNA	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
also	NN	O	O
investigated	NN	O	O
A-myb	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
49	NN	O	O
fresh	NN	O	O
cases	NN	O	O
of	NN	O	O
B	NN	O	O
leukemias	NN	O	O
.	NN	O	O

Among	NN	O	O
24	NN	O	O
ALL	NN	O	O
,	NN	O	O
6	NN	O	O
were	NN	O	O
of	NN	O	O
the	NN	O	O
null	NN	O	O
and	NN	O	O
11	NN	O	O
of	NN	O	O
the	NN	O	O
common	NN	O	O
type	NN	O	O
and	NN	O	O
all	NN	O	O
these	NN	O	O
were	NN	O	O
negative	NN	O	O
for	NN	O	O
A-myb	NN	O	B-protein
expression	NN	O	O
;	NN	O	O
on	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
all	NN	O	O
7	NN	O	O
B-ALL	NN	O	O
cases	NN	O	O
(	NN	O	O
slg+	NN	O	O
)	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
one	NN	O	O
fresh	NN	O	O
BL	NN	O	O
case	NN	O	O
with	NN	O	O
bone	NN	O	O
marrow	NN	O	O
infiltration	NN	O	O
,	NN	O	O
expressed	NN	O	O
A-myb	NN	O	B-DNA
.	NN	O	O

A-myb	NN	O	B-DNA
was	NN	O	O
undetectable	NN	O	O
in	NN	O	O
4	NN	O	O
prolymphocytic	NN	O	O
leukemias	NN	O	O
(	NN	O	O
PLL	NN	O	O
)	NN	O	O
but	NN	O	O
was	NN	O	O
strongly	NN	O	O
expressed	NN	O	O
in	NN	O	O
5/20	NN	O	O
(	NN	O	O
25	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CLL	NN	O	O
)	NN	O	O
samples	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
latter	NN	O	O
A-myb	NN	O	B-DNA
did	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
phenotype	NN	O	O
or	NN	O	O
clinical	NN	O	O
stage	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
one	NN	O	O
case	NN	O	O
of	NN	O	O
CLL	NN	O	O
into	NN	O	O
Richter	NN	O	O
's	NN	O	O
syndrome	NN	O	O
and	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
Richter	NN	O	B-cell_line
's	NN	O	I-cell_line
cells	NN	O	I-cell_line
expressed	NN	O	O
about	NN	O	O
25-fold	NN	O	O
less	NN	O	O
A-myb	NN	O	B-RNA
RNA	NN	O	I-RNA
than	NN	O	O
the	NN	O	O
CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
same	NN	O	O
patient.	NN	O	O
The	NN	O	O
pattern	NN	O	O
of	NN	O	O
c-myb	NN	O	B-DNA
and	NN	O	O
B-myb	NN	O	B-DNA
was	NN	O	O
clearly	NN	O	O
distinct	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
A-myb	NN	O	B-DNA
.	NN	O	O

C-myb	NN	O	O
and	NN	O	O
B-myb	NN	O	B-DNA
were	NN	O	O
expressed	NN	O	O
in	NN	O	O
all	NN	O	O
neoplastic	NN	O	O
groups	NN	O	O
,	NN	O	O
except	NN	O	O
in	NN	O	O
CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
A-myb	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
unlike	NN	O	O
that	NN	O	O
of	NN	O	O
c-myb	NN	O	B-DNA
and	NN	O	O
B-myb	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
B-cell	NN	O	O
neoplasias	NN	O	O
(	NN	O	O
in	NN	O	O
particular	NN	O	O
BL	NN	O	O
and	NN	O	O
slg+B-ALL	NN	O	O
)	NN	O	O
representative	NN	O	O
of	NN	O	O
a	NN	O	O
specific	NN	O	O
stage	NN	O	O
of	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

This	NN	O	O
expression	NN	O	O
may	NN	O	O
in	NN	O	O
part	NN	O	O
reflect	NN	O	O
expression	NN	O	O
of	NN	O	O
A-myb	NN	O	B-DNA
by	NN	O	O
the	NN	O	O
normal	NN	O	O
germinal	NN	O	O
center	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
are	NN	O	O
the	NN	O	O
normal	NN	O	O
counterpart	NN	O	O
of	NN	O	O
these	NN	O	O
transformed	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
data	NN	O	O
presented	NN	O	O
strongly	NN	O	O
support	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
perhaps	NN	O	O
in	NN	O	O
B-cell	NN	O	O
transformation	NN	O	O
in	NN	O	O
some	NN	O	O
neoplasias	NN	O	O
.	NN	O	O

-DOCSTART-	O

Granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
stimulates	NN	O	O
JAK2	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
and	NN	O	O
rapidly	NN	O	O
activates	NN	O	O
p93fes	NN	O	B-protein
,	NN	O	O
STAT1	NN	O	B-protein
p91	NN	O	I-protein
,	NN	O	O
and	NN	O	O
STAT3	NN	O	B-protein
p92	NN	O	I-protein
in	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

Granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
supports	NN	O	O
proliferation	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
functional	NN	O	O
activation	NN	O	O
of	NN	O	O
hemopoietic	NN	O	O
cells	NN	O	O
by	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
a	NN	O	O
heterodimeric	NN	O	O
receptor	NN	O	O
.	NN	O	O

Although	NN	O	O
GM-CSF	NN	O	B-protein
receptor	NN	O	O
is	NN	O	O
devoid	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
enzymatic	NN	O	O
activity	NN	O	O
,	NN	O	O
GM-CSF-induced	NN	O	B-cell_line
peripheral	NN	O	I-cell_line
blood	NN	O	I-cell_line
polymorphonuclear	NN	O	I-cell_line
leukocytes	NN	O	I-cell_line
(	NN	O	O
PMN	NN	O	B-cell_type
)	NN	O	O
functional	NN	O	O
activation	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
a	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
intracellular	NN	O	B-protein
signaling	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
JAK2	NN	O	B-protein
becomes	NN	O	O
tyrosine-phosphorylated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
GM-CSF	NN	O	B-protein
in	NN	O	O
PMN	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
also	NN	O	O
the	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
STAT1	NN	O	B-protein
p91	NN	O	I-protein
and	NN	O	O
STAT3	NN	O	B-protein
p92	NN	O	I-protein
and	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
c-fps/fes	NN	O	B-DNA
protooncogene	NN	O	I-DNA
become	NN	O	O
tyrosine-phosphorylated	NN	O	O
upon	NN	O	O
GM-CSF	NN	O	B-protein
stimulation	NN	O	O
and	NN	O	O
physically	NN	O	O
associated	NN	O	O
with	NN	O	O
both	NN	O	O
GM-CSF	NN	O	B-protein
receptor	NN	O	O
beta	NN	O	O
common	NN	O	O
subunit	NN	O	O
and	NN	O	O
JAK2	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
GM-CSF	NN	O	B-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
JAK2	NN	O	B-protein
and	NN	O	O
p93fes	NN	O	B-protein
catalytic	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-protein
receptor	NN	O	I-protein
beta	NN	O	I-protein
common	NN	O	I-protein
subunit	NN	O	I-protein
with	NN	O	O
JAK2	NN	O	B-protein
is	NN	O	O
ligand-dependent	NN	O	O
.	NN	O	O

Finally	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
GM-CSF	NN	O	B-protein
induces	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
complex	NN	O	I-protein
that	NN	O	O
contains	NN	O	O
both	NN	O	O
p91	NN	O	B-protein
and	NN	O	O
p92	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
identify	NN	O	O
a	NN	O	O
new	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
activated	NN	O	O
by	NN	O	O
GM-CSF	NN	O	B-protein
and	NN	O	O
provide	NN	O	O
a	NN	O	O
mechanism	NN	O	O
for	NN	O	O
rapid	NN	O	O
activation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
GM-CSF-stimulated	NN	O	B-cell_type
PMN	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	I-protein
EBNA2	NN	O	I-protein
)	NN	O	I-protein
-oestrogen	NN	O	I-protein
receptor	NN	O	I-protein
fusion	NN	O	I-protein
proteins	NN	O	I-protein
complement	NN	O	O
the	NN	O	O
EBNA2-deficient	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
strain	NN	O	O
P3HR1	NN	O	O
in	NN	O	O
transformation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
suppress	NN	O	O
growth	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
develop	NN	O	O
a	NN	O	O
transformation	NN	O	O
system	NN	O	O
with	NN	O	O
a	NN	O	O
conditional	NN	O	O
Epstein-Barr	NN	O	B-DNA
virus	NN	O	I-DNA
nuclear	NN	O	I-DNA
antigen	NN	O	I-DNA
2	NN	O	I-DNA
(	NN	O	I-DNA
EBNA2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
fused	NN	O	O
the	NN	O	O
hormone	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
oestrogen	NN	O	O
receptor	NN	O	O
to	NN	O	O
the	NN	O	O
N	NN	O	B-protein
or	NN	O	I-protein
C	NN	O	I-protein
terminus	NN	O	I-protein
of	NN	O	O
EBNA2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
promoter	NN	O	O
transactivation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
primary	NN	O	O
B	NN	O	O
cell	NN	O	O
transformation	NN	O	O
assays	NN	O	O
these	NN	O	O
chimeric	NN	O	O
EBNA2	NN	O	B-protein
proteins	NN	O	O
are	NN	O	O
able	NN	O	O
to	NN	O	O
substitute	NN	O	O
for	NN	O	O
wild-type	NN	O	B-protein
EBNA2	NN	O	I-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
oestrogen	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
this	NN	O	O
transformation	NN	O	O
is	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
double	NN	O	O
infection	NN	O	O
of	NN	O	O
a	NN	O	O
cell	NN	O	O
with	NN	O	O
two	NN	O	O
virions	NN	O	O
,	NN	O	O
the	NN	O	O
P3HR1	NN	O	B-DNA
virus	NN	O	I-DNA
genome	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
mini-EBV	NN	O	B-DNA
plasmid	NN	O	I-DNA
carrying	NN	O	O
the	NN	O	O
chimeric	NN	O	O
EBNA2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
EBNA2	NN	O	B-protein
-oestrogen	NN	O	O
receptor	NN	O	O
fusion	NN	O	O
protein	NN	O	O
in	NN	O	O
established	NN	O	B-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
lines	NN	O	I-cell_line
resulted	NN	O	O
in	NN	O	O
growth	NN	O	O
retardation	NN	O	O
or	NN	O	O
growth	NN	O	O
arrest	NN	O	O
upon	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
oestrogen	NN	O	O
.	NN	O	O

By	NN	O	O
titrating	NN	O	O
the	NN	O	O
oestrogen	NN	O	O
concentration	NN	O	O
in	NN	O	O
these	NN	O	O
stably	NN	O	B-cell_line
transfected	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
growth	NN	O	O
retarding	NN	O	O
and	NN	O	O
the	NN	O	O
transactivating	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
proteins	NN	O	I-protein
could	NN	O	O
not	NN	O	O
be	NN	O	O
dissociated	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
of	NN	O	O
established	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
lines	NN	O	I-cell_line
is	NN	O	O
a	NN	O	O
novel	NN	O	O
function	NN	O	O
of	NN	O	O
EBNA2	NN	O	B-protein
which	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	O
system	NN	O	O
.	NN	O	O

It	NN	O	O
remains	NN	O	O
open	NN	O	O
whether	NN	O	O
the	NN	O	O
growth	NN	O	O
retarding	NN	O	O
property	NN	O	O
of	NN	O	O
the	NN	O	O
EBNA2-oestrogen	NN	O	B-protein
receptor	NN	O	I-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
lines	NN	O	I-cell_line
is	NN	O	O
due	NN	O	O
to	NN	O	O
unphysiologically	NN	O	O
high	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
protein	NN	O	I-protein
or	NN	O	O
to	NN	O	O
interference	NN	O	O
with	NN	O	O
a	NN	O	O
cellular	NN	O	O
programme	NN	O	O
driving	NN	O	O
proliferation	NN	O	O
in	NN	O	O
these	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Tissue-specific	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
gammac	NN	O	B-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
depends	NN	O	O
upon	NN	O	O
an	NN	O	O
Ets	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
GA-binding	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
gammac	NN	O	B-protein
chain	NN	O	I-protein
is	NN	O	O
a	NN	O	O
subunit	NN	O	O
of	NN	O	O
multiple	NN	O	O
cytokine	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-7	NN	O	B-protein
,	NN	O	O
IL-9	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-15	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
hematopoietic	NN	O	B-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
defect	NN	O	O
in	NN	O	O
gammac	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
X-linked	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
block	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
better	NN	O	O
characterize	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
gammac	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
define	NN	O	O
the	NN	O	O
minimal	NN	O	B-DNA
tissue-specific	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
,	NN	O	O
progressive	NN	O	O
5'-deletion	NN	O	B-DNA
constructs	NN	O	I-DNA
of	NN	O	O
a	NN	O	O
segment	NN	O	O
extending	NN	O	O
1053	NN	O	B-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
transcription	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
were	NN	O	O
generated	NN	O	O
and	NN	O	O
tested	NN	O	O
for	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
various	NN	O	O
hematopoietic	NN	O	B-cell_type
and	NN	O	I-cell_type
nonhematopoietic	NN	O	I-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
-1053/+34	NN	O	B-DNA
construct	NN	O	I-DNA
allowed	NN	O	O
promoter	NN	O	O
activity	NN	O	O
only	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
origin	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
tissue	NN	O	O
specificity	NN	O	O
was	NN	O	O
conserved	NN	O	O
in	NN	O	O
all	NN	O	O
other	NN	O	O
constructs	NN	O	O
tested	NN	O	O
.	NN	O	O

The	NN	O	O
region	NN	O	O
downstream	NN	O	O
of	NN	O	O
-90	NN	O	O
appeared	NN	O	O
critical	NN	O	O
for	NN	O	O
basal	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

It	NN	O	O
contains	NN	O	O
two	NN	O	O
potential	NN	O	O
Ets	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
conserved	NN	O	O
in	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
gammac	NN	O	I-DNA
promoter	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
was	NN	O	O
found	NN	O	O
essential	NN	O	O
for	NN	O	O
functional	NN	O	O
promoter	NN	O	O
activity	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
mutational	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
functional	NN	O	O
Ets	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
found	NN	O	O
to	NN	O	O
bind	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
principally	NN	O	O
GA-binding	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
Elf-1	NN	O	B-protein
and	NN	O	O
could	NN	O	O
be	NN	O	O
transactivated	NN	O	O
by	NN	O	O
GABPalpha	NN	O	B-protein
and	NN	O	O
-beta	NN	O	B-protein
synergistically	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
,	NN	O	O
as	NN	O	O
already	NN	O	O
reported	NN	O	O
for	NN	O	O
the	NN	O	O
IL2Rbeta	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
GA-binding	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
an	NN	O	O
essential	NN	O	O
component	NN	O	O
of	NN	O	O
gammac	NN	O	O
basal	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Although	NN	O	O
GABP	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
not	NN	O	O
restricted	NN	O	O
to	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
other	NN	O	O
specific	NN	O	B-protein
factors	NN	O	I-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gammac	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Detection	NN	O	O
of	NN	O	O
intracellular	NN	O	B-protein
signal	NN	O	I-protein
transduction	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
rhesus	NN	O	O
macaques	NN	O	O
and	NN	O	O
sooty	NN	O	O
mangabeys	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
manifestations	NN	O	O
of	NN	O	O
human	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
nonhuman	NN	O	O
primate	NN	O	O
SIV	NN	O	O
infection	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
disease	NN	O	O
is	NN	O	O
reasoned	NN	O	O
to	NN	O	O
be	NN	O	O
secondary	NN	O	O
to	NN	O	O
generalized	NN	O	O
T-cell	NN	O	O
dysfunction	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
T-cell	NN	O	O
dysfunction	NN	O	O
remain	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O

To	NN	O	O
address	NN	O	O
this	NN	O	O
issue	NN	O	O
,	NN	O	O
we	NN	O	O
sought	NN	O	O
to	NN	O	O
utilize	NN	O	O
the	NN	O	O
nonhuman	NN	O	O
primate	NN	O	O
model	NN	O	O
to	NN	O	O
study	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
events	NN	O	O
in	NN	O	O
cells	NN	O	O
from	NN	O	O
disease-susceptible	NN	O	O
rhesus	NN	O	O
macaques	NN	O	O
and	NN	O	O
disease-resistant	NN	O	O
sooty	NN	O	O
mangabeys	NN	O	O
.	NN	O	O

Because	NN	O	O
relatively	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
these	NN	O	O
events	NN	O	O
in	NN	O	O
nonhuman	NN	O	O
primates	NN	O	O
,	NN	O	O
our	NN	O	O
laboratory	NN	O	O
defined	NN	O	O
optimal	NN	O	O
conditions	NN	O	O
,	NN	O	O
reagents	NN	O	O
,	NN	O	O
and	NN	O	O
assays	NN	O	O
for	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
in	NN	O	O
cells	NN	O	O
from	NN	O	O
nonhuman	NN	O	O
primates	NN	O	O
.	NN	O	O

The	NN	O	O
protein	NN	O	O
phosphorylation	NN	O	O
patterns	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
monkeys	NN	O	O
exhibited	NN	O	O
quantitative	NN	O	O
,	NN	O	O
qualitative	NN	O	O
,	NN	O	O
and	NN	O	O
kinetic	NN	O	O
differences	NN	O	O
.	NN	O	O

Antibodies	NN	O	O
to	NN	O	O
Stat6	NN	O	B-protein
detected	NN	O	O
a	NN	O	O
unique	NN	O	O
band	NN	O	O
in	NN	O	O
macaque	NN	O	O
cell	NN	O	O
lysates	NN	O	O
.	NN	O	O

This	NN	O	O
band	NN	O	O
is	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
human	NN	O	O
cell	NN	O	O
lysates	NN	O	O
and	NN	O	O
never	NN	O	O
seen	NN	O	O
in	NN	O	O
mangabey	NN	O	O
cell	NN	O	O
lysates	NN	O	O
.	NN	O	O

Detection	NN	O	O
of	NN	O	O
various	NN	O	O
other	NN	O	O
intracellular	NN	O	B-protein
signaling	NN	O	I-protein
proteins	NN	O	I-protein
is	NN	O	O
also	NN	O	O
described	NN	O	O
.	NN	O	O

-DOCSTART-	O

Genetic	NN	O	O
characterization	NN	O	O
of	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
T-cell	NN	O	I-DNA
leukemia	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
promoter	NN	O	I-DNA
:	NN	O	O
Binding	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
to	NN	O	O
Tax-responsive	NN	O	B-DNA
element	NN	O	I-DNA
1	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
cyclic	NN	O	B-protein
AMP-responsive	NN	O	I-protein
members	NN	O	I-protein
of	NN	O	O
the	NN	O	O
CREB/ATF	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	O
factors	NN	O	O
.	NN	O	O

To	NN	O	O
achieve	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
transactivation	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
Tax-responsive	NN	O	B-DNA
element	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	O
TRE-1	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
we	NN	O	O
developed	NN	O	O
a	NN	O	O
genetic	NN	O	O
approach	NN	O	O
with	NN	O	O
Saccharomyces	NN	O	O
cerevisiae	NN	O	O
.	NN	O	O

We	NN	O	O
constructed	NN	O	O
a	NN	O	O
yeast	NN	O	O
reporter	NN	O	O
strain	NN	O	O
containing	NN	O	O
the	NN	O	O
lacZ	NN	O	B-DNA
gene	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
CYC1	NN	O	B-DNA
promoter	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
three	NN	O	O
copies	NN	O	O
of	NN	O	O
TRE-1	NN	O	B-DNA
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
cyclic	NN	O	B-protein
AMP	NN	O	I-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
or	NN	O	O
CREB	NN	O	B-protein
fused	NN	O	O
to	NN	O	O
the	NN	O	O
GAL4	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
(	NN	O	O
GAD	NN	O	B-protein
)	NN	O	O
in	NN	O	O
this	NN	O	O
strain	NN	O	O
did	NN	O	O
not	NN	O	O
modify	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Tax	NN	O	B-protein
alone	NN	O	O
was	NN	O	O
also	NN	O	O
inactive	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
induced	NN	O	O
by	NN	O	O
coexpression	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
and	NN	O	O
CREB	NN	O	B-protein
.	NN	O	O

This	NN	O	O
effect	NN	O	O
was	NN	O	O
stronger	NN	O	O
with	NN	O	O
the	NN	O	O
GAD-CREB	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
different	NN	O	O
CREB	NN	O	B-protein
mutants	NN	O	I-protein
with	NN	O	O
this	NN	O	O
genetic	NN	O	O
system	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
92	NN	O	I-protein
amino	NN	O	I-protein
acid	NN	O	I-protein
residues	NN	O	I-protein
,	NN	O	O
which	NN	O	O
include	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
domain	NN	O	I-protein
and	NN	O	O
the	NN	O	O
leucine	NN	O	B-protein
zipper	NN	O	I-protein
,	NN	O	O
are	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
to	NN	O	O
mediate	NN	O	O
transactivation	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

To	NN	O	O
identify	NN	O	O
cellular	NN	O	B-protein
proteins	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
TRE-1	NN	O	B-DNA
in	NN	O	O
a	NN	O	O
Tax	NN	O	B-protein
-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
this	NN	O	O
strain	NN	O	O
was	NN	O	O
also	NN	O	O
used	NN	O	O
to	NN	O	O
screen	NN	O	O
a	NN	O	O
library	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
cDNAs	NN	O	I-DNA
fused	NN	O	O
to	NN	O	O
GAD	NN	O	B-protein
.	NN	O	O

Of	NN	O	O
five	NN	O	O
positive	NN	O	O
clones	NN	O	O
isolated	NN	O	O
from	NN	O	O
0.75	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
yeast	NN	O	O
colonies	NN	O	O
,	NN	O	O
four	NN	O	O
were	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
CREB/activating	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
ATF	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
:	NN	O	O
CREB	NN	O	B-protein
,	NN	O	O
two	NN	O	B-protein
isoforms	NN	O	I-protein
of	NN	O	I-protein
the	NN	O	I-protein
cyclic	NN	O	I-protein
AMP-responsive	NN	O	I-protein
element	NN	O	I-protein
modulator	NN	O	I-protein
(	NN	O	O
CREM	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
ATF-1	NN	O	B-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
these	NN	O	O
three	NN	O	O
proteins	NN	O	O
can	NN	O	O
be	NN	O	O
phosphorylated	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
and	NN	O	O
thus	NN	O	O
form	NN	O	O
a	NN	O	O
particular	NN	O	O
subgroup	NN	O	O
within	NN	O	O
the	NN	O	O
CREB/ATF	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
ATF-2	NN	O	B-protein
in	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
did	NN	O	O
not	NN	O	O
activate	NN	O	O
TRE-1	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

This	NN	O	O
shows	NN	O	O
that	NN	O	O
in	NN	O	O
a	NN	O	O
eukaryotic	NN	O	O
nucleus	NN	O	O
,	NN	O	O
Tax	NN	O	B-protein
specifically	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
domain-leucine	NN	O	I-protein
zipper	NN	O	I-protein
region	NN	O	I-protein
of	NN	O	O
ATF-1	NN	O	B-protein
,	NN	O	O
CREB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CREM	NN	O	B-protein
.	NN	O	O

The	NN	O	O
fifth	NN	O	O
clone	NN	O	O
identified	NN	O	O
in	NN	O	O
this	NN	O	O
screening	NN	O	O
corresponded	NN	O	O
to	NN	O	O
the	NN	O	O
Ku	NN	O	B-protein
autoantigen	NN	O	I-protein
p70	NN	O	I-protein
subunit	NN	O	I-protein
.	NN	O	O

When	NN	O	O
fused	NN	O	O
to	NN	O	O
GAD	NN	O	B-protein
,	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
Ku	NN	O	B-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
via	NN	O	O
TRE-1	NN	O	B-DNA
but	NN	O	O
this	NN	O	O
activation	NN	O	O
was	NN	O	O
not	NN	O	O
dependent	NN	O	O
on	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
mammalian	NN	O	B-protein
histone	NN	O	I-protein
deacetylase	NN	O	I-protein
related	NN	O	O
to	NN	O	O
the	NN	O	O
yeast	NN	O	B-protein
transcriptional	NN	O	I-protein
regulator	NN	O	I-protein
Rpd3p	NN	O	B-protein
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Trapoxin	NN	O	O
is	NN	O	O
a	NN	O	O
microbially	NN	O	O
derived	NN	O	O
cyclotetrapeptide	NN	O	O
that	NN	O	O
inhibits	NN	O	O
histone	NN	O	B-protein
deacetylation	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
causes	NN	O	O
mammalian	NN	O	O
cells	NN	O	O
to	NN	O	O
arrest	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

A	NN	O	O
trapoxin	NN	O	O
affinity	NN	O	O
matrix	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
isolate	NN	O	O
two	NN	O	O
nuclear	NN	O	O
proteins	NN	O	O
that	NN	O	O
copurified	NN	O	O
with	NN	O	O
histone	NN	O	B-protein
deacetylase	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

Both	NN	O	O
proteins	NN	O	O
were	NN	O	O
identified	NN	O	O
by	NN	O	O
peptide	NN	O	O
microsequencing	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
complementary	NN	O	B-DNA
DNA	NN	O	I-DNA
encoding	NN	O	O
the	NN	O	O
histone	NN	O	B-protein
deacetylase	NN	O	I-protein
catalytic	NN	O	I-protein
subunit	NN	O	I-protein
(	NN	O	O
HD1	NN	O	B-protein
)	NN	O	O
was	NN	O	O
cloned	NN	O	O
from	NN	O	O
a	NN	O	O
human	NN	O	O
Jurkat	NN	O	O
T	NN	O	O
cell	NN	O	O
library	NN	O	O
.	NN	O	O

As	NN	O	O
the	NN	O	O
predicted	NN	O	O
protein	NN	O	O
is	NN	O	O
very	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
yeast	NN	O	B-protein
transcriptional	NN	O	I-protein
regulator	NN	O	I-protein
Rpd3p	NN	O	B-protein
,	NN	O	O
these	NN	O	O
results	NN	O	O
support	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
histone	NN	O	B-protein
deacetylase	NN	O	I-protein
as	NN	O	O
a	NN	O	O
key	NN	O	O
regulator	NN	O	O
of	NN	O	O
eukaryotic	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Coexpression	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-13	NN	O	B-DNA
and	NN	O	I-DNA
interleukin-4	NN	O	I-DNA
genes	NN	O	I-DNA
correlates	NN	O	O
with	NN	O	O
their	NN	O	O
physical	NN	O	O
linkage	NN	O	O
in	NN	O	O
the	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
cluster	NN	O	I-DNA
on	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
5q23-31	NN	O	I-DNA
.	NN	O	O

Interleukin-13	NN	O	B-protein
(	NN	O	O
IL-13	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IL-4	NN	O	B-protein
are	NN	O	O
cytokines	NN	O	O
produced	NN	O	O
by	NN	O	O
T	NN	O	O
cells	NN	O	O
that	NN	O	O
are	NN	O	O
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
q23-31	NN	O	O
region	NN	O	O
of	NN	O	O
human	NN	O	O
chromosome	NN	O	O
5	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-13	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
and	NN	O	O
sequenced	NN	O	O
the	NN	O	O
human	NN	O	O
IL-13	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
analyzed	NN	O	O
its	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
for	NN	O	O
potential	NN	O	O
transcriptional	NN	O	B-DNA
activation	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
examined	NN	O	O
its	NN	O	O
expression	NN	O	O
in	NN	O	O
nontransformed	NN	O	B-cell_line
T-lineage	NN	O	I-cell_line
cell	NN	O	I-cell_line
populations	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
IL-13	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
located	NN	O	O
12.5-kb	NN	O	B-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
2-kb	NN	O	B-DNA
downstream	NN	O	I-DNA
of	NN	O	O
a	NN	O	O
CpG	NN	O	B-DNA
island	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
IL-13	NN	O	B-DNA
gene	NN	O	I-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
flank	NN	O	I-DNA
region	NN	O	I-DNA
included	NN	O	O
a	NN	O	O
segment	NN	O	O
with	NN	O	O
sequence	NN	O	O
homology	NN	O	O
to	NN	O	O
P	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-13	NN	O	B-DNA
P	NN	O	I-DNA
element	NN	O	I-DNA
site	NN	O	I-DNA
significantly	NN	O	O
reduced	NN	O	O
IL-13	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Oligonucleotides	NN	O	O
containing	NN	O	O
the	NN	O	O
IL-13	NN	O	B-protein
or	NN	O	O
IL-4	NN	O	B-DNA
P	NN	O	I-DNA
element	NN	O	I-DNA
sites	NN	O	I-DNA
specifically	NN	O	O
bound	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
nuclear	NN	O	B-protein
factor-activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
,	NN	O	O
preformed	NN	O	O
(	NN	O	O
NF-ATp	NN	O	B-protein
)	NN	O	O
,	NN	O	O
when	NN	O	O
incubated	NN	O	O
with	NN	O	O
nuclear	NN	O	O
protein	NN	O	O
extracts	NN	O	O
from	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-13	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
was	NN	O	O
highest	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_line
populations	NN	O	I-cell_line
enriched	NN	O	O
for	NN	O	O
cells	NN	O	O
that	NN	O	O
had	NN	O	O
previously	NN	O	O
been	NN	O	O
primed	NN	O	O
in	NN	O	O
vivo	NN	O	O
or	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
priming	NN	O	O
increases	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-13	NN	O	B-DNA
and	NN	O	I-DNA
IL-4	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
coordinate	NN	O	O
manner	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
primed	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
contain	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
NF-ATp	NN	O	I-protein
,	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
P	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
and	NN	O	I-DNA
IL-13	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
than	NN	O	O
do	NN	O	O
freshly-isolated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
NF-AT-binding	NN	O	B-DNA
P	NN	O	I-DNA
elements	NN	O	I-DNA
are	NN	O	O
attractive	NN	O	O
candidates	NN	O	O
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
coordinate	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
alpha	NN	O	B-protein
interferon	NN	O	I-protein
but	NN	O	O
not	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
signal	NN	O	O
transduction	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
tyrosine	NN	O	B-protein
phosphatase	NN	O	I-protein
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
viral	NN	O	O
oncoproteins	NN	O	O
,	NN	O	O
cell	NN	O	O
transformation	NN	O	O
,	NN	O	O
or	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
can	NN	O	O
inhibit	NN	O	O
alpha/beta	NN	O	O
interferon	NN	O	O
(	NN	O	O
IFN-alpha/beta	NN	O	O
)	NN	O	O
-induced	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
these	NN	O	O
promoters	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
exert	NN	O	O
their	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
vary	NN	O	O
,	NN	O	O
but	NN	O	O
in	NN	O	O
most	NN	O	O
instances	NN	O	O
they	NN	O	O
involve	NN	O	O
a	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-alpha/beta-induced	NN	O	O
transcription	NN	O	O
complex	NN	O	O
ISGF3	NN	O	O
such	NN	O	O
that	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
component	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
(	NN	O	O
the	NN	O	O
48-kDa	NN	O	O
ISGF3gamma	NN	O	O
protein	NN	O	O
)	NN	O	O
does	NN	O	O
not	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
interferon-stimulated	NN	O	O
response	NN	O	O
element	NN	O	O
(	NN	O	O
ISRE	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
treatment	NN	O	O
of	NN	O	O
human	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
monocytes	NN	O	O
dramatically	NN	O	O
inhibits	NN	O	O
activation	NN	O	O
of	NN	O	O
IFN-alpha/B-stimulated	NN	O	O
early	NN	O	O
response	NN	O	O
genes	NN	O	O
but	NN	O	O
by	NN	O	O
a	NN	O	O
mechanism	NN	O	O
which	NN	O	O
does	NN	O	O
not	NN	O	O
involve	NN	O	O
abrogation	NN	O	O
of	NN	O	O
the	NN	O	O
ISRE	NN	O	O
binding	NN	O	O
of	NN	O	O
ISGF3gamma	NN	O	O
.	NN	O	O

Phorbol	NN	O	O
ester	NN	O	O
treatment	NN	O	O
of	NN	O	O
monocytes	NN	O	O
inhibited	NN	O	O
IFN	NN	O	O
alpha-stimulated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
Stat1alpha	NN	O	B-protein
,	NN	O	O
Stat2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Stat3	NN	O	B-protein
and	NN	O	O
of	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
Tyk2	NN	O	B-protein
but	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
IFN-gamma	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
Stat1alpha	NN	O	B-protein
.	NN	O	O

IFNalpha-stimulated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
IFN-alpha	NN	O	B-protein
receptor	NN	O	I-protein
were	NN	O	O
unaffected	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
PMA	NN	O	O
caused	NN	O	O
the	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
Tyk2	NN	O	B-protein
but	NN	O	O
not	NN	O	O
of	NN	O	O
Jak1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
was	NN	O	O
activated	NN	O	O
by	NN	O	O
IFN	NN	O	B-protein
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
vanadate	NN	O	O
prevented	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
PMA	NN	O	O
with	NN	O	O
regard	NN	O	O
to	NN	O	O
PMA-induced	NN	O	O
Tyk2	NN	O	B-protein
dephosphorylation	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
PMA	NN	O	O
exerts	NN	O	O
its	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
tyrosine	NN	O	B-protein
phosphatase	NN	O	I-protein
which	NN	O	O
selectively	NN	O	O
regulates	NN	O	O
Tyk2	NN	O	B-protein
but	NN	O	O
not	NN	O	O
Jak1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
normal	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Convergence	NN	O	O
of	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
and	NN	O	O
differences	NN	O	O
from	NN	O	O
transformed	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
study	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
have	NN	O	O
optimized	NN	O	O
conditions	NN	O	O
for	NN	O	O
transient	NN	O	O
transfection	NN	O	O
.	NN	O	O

Interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
-reporter	NN	O	B-DNA
gene	NN	O	I-DNA
behavior	NN	O	O
closely	NN	O	O
parallels	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
.	NN	O	O

As	NN	O	O
assessed	NN	O	O
with	NN	O	O
mutagenized	NN	O	B-DNA
promoters	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
most	NN	O	O
important	NN	O	O
IL-2	NN	O	B-DNA
cis-regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
AP-1	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
NF-	NN	O	B-DNA
kappaB	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
primary	NN	O	O
activation	NN	O	O
,	NN	O	O
with	NN	O	O
phytohemagglutinin	NN	O	B-protein
or	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	I-protein
CD3	NN	O	I-protein
,	NN	O	O
and	NN	O	O
costimulation	NN	O	O
,	NN	O	O
provided	NN	O	O
by	NN	O	O
pairs	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
antibodies	NN	O	I-protein
or	NN	O	O
B7-positive	NN	O	B-cell_type
(	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
)	NN	O	I-cell_type
or	NN	O	O
B7-negative	NN	O	B-cell_type
(	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
)	NN	O	I-cell_type
accessory	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	I-cell_type
are	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
same	NN	O	O
cis-elements	NN	O	B-DNA
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
of	NN	O	I-DNA
activated	NN	O	I-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
much	NN	O	O
less	NN	O	O
important	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
than	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
transcriptional	NN	O	O
regulation	NN	O	O
differs	NN	O	O
in	NN	O	O
tumor	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
compared	NN	O	O
with	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
different	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
converge	NN	O	O
on	NN	O	O
the	NN	O	O
same	NN	O	O
cis-elements	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
RAR-RXR	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
RXR-RXR	NN	O	B-protein
pathway	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
signaling	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
CD34+	NN	O	I-cell_line
erythroid	NN	O	I-cell_line
progenitor	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
,	NN	O	O
similar	NN	O	O
to	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
can	NN	O	O
act	NN	O	O
as	NN	O	O
a	NN	O	O
bifunctional	NN	O	O
regulator	NN	O	O
of	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
,	NN	O	O
in	NN	O	O
that	NN	O	O
it	NN	O	O
can	NN	O	O
stimulate	NN	O	O
granulocyte-macrophage	NN	O	O
colony-stimulating	NN	O	O
factor	NN	O	O
(	NN	O	O
GM-CSF	NN	O	O
)	NN	O	O
-	NN	O	O
or	NN	O	O
interleukin-3	NN	O	O
(	NN	O	O
IL-3	NN	O	O
)	NN	O	O
-induced	NN	O	O
GM	NN	O	O
colony	NN	O	O
formation	NN	O	O
,	NN	O	O
but	NN	O	O
potently	NN	O	O
inhibit	NN	O	O
G-CSF	NN	O	B-protein
-induced	NN	O	O
growth	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
using	NN	O	O
highly	NN	O	O
enriched	NN	O	O
human	NN	O	B-protein
CD34+	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
Lin-	NN	O	B-cell_line
murine	NN	O	I-cell_line
bone	NN	O	I-cell_line
marrow	NN	O	I-cell_line
progenitor	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
demonstrates	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
9-cis-RA	NN	O	O
on	NN	O	O
burst-forming	NN	O	O
unit-erythroid	NN	O	O
(	NN	O	O
BFU-E	NN	O	O
)	NN	O	O
colony	NN	O	O
formation	NN	O	O
regardless	NN	O	O
of	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
stimulating	NN	O	O
growth	NN	O	O
.	NN	O	O

Specifically	NN	O	O
,	NN	O	O
9-cis-RA	NN	O	O
potently	NN	O	O
inhibited	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
BFU-E	NN	O	O
response	NN	O	O
to	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
Epo	NN	O	B-protein
)	NN	O	O
(	NN	O	O
100	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
SCF	NN	O	B-protein
)	NN	O	O
+	NN	O	O
Epo	NN	O	B-protein
(	NN	O	O
92	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
IL-3	NN	O	B-protein
+	NN	O	O
Epo	NN	O	B-protein
(	NN	O	O
97	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
+	NN	O	O
Epo	NN	O	B-protein
(	NN	O	O
88	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
IL-9	NN	O	B-protein
+	NN	O	O
Epo	NN	O	B-protein
(	NN	O	O
100	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

Erythroid	NN	O	O
colony	NN	O	O
growth	NN	O	O
was	NN	O	O
also	NN	O	O
inhibited	NN	O	O
when	NN	O	O
CD34+	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
were	NN	O	O
seeded	NN	O	O
at	NN	O	O
one	NN	O	O
cell	NN	O	O
per	NN	O	O
well	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
direct	NN	O	O
action	NN	O	O
of	NN	O	O
RA	NN	O	O
.	NN	O	O

Using	NN	O	O
synthetic	NN	O	O
ligands	NN	O	O
to	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RARs	NN	O	B-protein
)	NN	O	O
and	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RXRs	NN	O	B-protein
)	NN	O	O
that	NN	O	O
selectively	NN	O	O
bind	NN	O	O
and	NN	O	O
activate	NN	O	O
RAR-RXR	NN	O	B-protein
or	NN	O	O
RXR-RXR	NN	O	B-protein
dimers	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
we	NN	O	O
dissected	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
retinoid	NN	O	O
response	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
normal	NN	O	O
myeloid	NN	O	O
and	NN	O	O
erythroid	NN	O	O
progenitor	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

Transactivation	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
both	NN	O	O
the	NN	O	O
RAR	NN	O	B-protein
(	NN	O	O
Ro	NN	O	O
13-7410	NN	O	O
)	NN	O	O
and	NN	O	O
RXR	NN	O	B-protein
(	NN	O	O
Ro	NN	O	O
25-6603	NN	O	O
and	NN	O	O
Ro	NN	O	O
25-7386	NN	O	O
)	NN	O	O
ligands	NN	O	O
were	NN	O	O
highly	NN	O	O
selective	NN	O	O
at	NN	O	O
100	NN	O	O
nmol/L	NN	O	O
.	NN	O	O

At	NN	O	O
this	NN	O	O
concentration	NN	O	O
,	NN	O	O
Ro	NN	O	O
13-7410	NN	O	O
potently	NN	O	O
inhibited	NN	O	O
G-CSF	NN	O	B-protein
-stimulated	NN	O	O
myeloid	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
SCF	NN	O	B-protein
+	NN	O	O
Epo	NN	O	B-protein
-induced	NN	O	O
erythroid	NN	O	O
colony	NN	O	O
growth	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
same	NN	O	O
concentration	NN	O	O
,	NN	O	O
Ro	NN	O	O
25-6603	NN	O	O
and	NN	O	O
Ro	NN	O	O
25-7386	NN	O	O
had	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
G-CSF	NN	O	B-protein
-induced	NN	O	O
colony	NN	O	O
formation	NN	O	O
,	NN	O	O
whereas	NN	O	O
they	NN	O	O
inhibited	NN	O	O
75	NN	O	O
%	NN	O	O
and	NN	O	O
53	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
of	NN	O	O
SCF	NN	O	B-protein
+	NN	O	O
Epo	NN	O	B-protein
-stimulated	NN	O	O
BFU-E	NN	O	O
colony	NN	O	O
growth	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
RAR-RXR	NN	O	B-protein
response	NN	O	O
pathway	NN	O	O
can	NN	O	O
signal	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
and	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
unique	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
RXR-RXR	NN	O	B-protein
pathway	NN	O	O
in	NN	O	O
mediating	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
but	NN	O	O
not	NN	O	O
myeloid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

AM580	NN	O	O
,	NN	O	O
a	NN	O	O
stable	NN	O	O
benzoic	NN	O	O
derivative	NN	O	O
of	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
,	NN	O	O
has	NN	O	O
powerful	NN	O	O
and	NN	O	O
selective	NN	O	O
cyto-differentiating	NN	O	O
effects	NN	O	O
on	NN	O	O
acute	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

All-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
is	NN	O	O
successfully	NN	O	O
used	NN	O	O
in	NN	O	O
the	NN	O	O
cyto-differentiating	NN	O	O
treatment	NN	O	O
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
.	NN	O	O

Paradoxically	NN	O	O
,	NN	O	O
APL	NN	O	B-cell_line
cells	NN	O	I-cell_line
express	NN	O	O
PML-RAR	NN	O	B-protein
,	NN	O	O
an	NN	O	O
aberrant	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
type	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
leukemia-specific	NN	O	B-DNA
t	NN	O	I-DNA
(	NN	O	I-DNA
15	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
chromosomal	NN	O	I-DNA
translocation	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
AM580	NN	O	O
,	NN	O	O
a	NN	O	O
stable	NN	O	O
retinobenzoic	NN	O	O
derivative	NN	O	O
originally	NN	O	O
synthesized	NN	O	O
as	NN	O	O
a	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
agonist	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
powerful	NN	O	O
inducer	NN	O	O
of	NN	O	O
granulocytic	NN	O	O
maturation	NN	O	O
in	NN	O	O
NB4	NN	O	B-cell_line
,	NN	O	O
an	NN	O	O
APL-derived	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
in	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
APL	NN	O	I-cell_type
blasts	NN	O	I-cell_type
.	NN	O	O

After	NN	O	O
treatment	NN	O	O
of	NN	O	O
APL	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
AM580	NN	O	O
either	NN	O	O
alone	NN	O	O
or	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
G-CSF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
compound	NN	O	O
induces	NN	O	O
granulocytic	NN	O	O
maturation	NN	O	O
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
determination	NN	O	O
of	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
leukocyte	NN	O	B-RNA
alkaline	NN	O	I-RNA
phosphatase	NN	O	I-RNA
,	NN	O	I-RNA
CD11b	NN	O	I-RNA
,	NN	O	I-RNA
CD33	NN	O	I-RNA
,	NN	O	I-RNA
and	NN	O	I-RNA
G-CSF	NN	O	I-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
at	NN	O	O
concentrations	NN	O	O
that	NN	O	O
are	NN	O	O
10-	NN	O	O
to	NN	O	O
100-fold	NN	O	O
lower	NN	O	O
than	NN	O	O
those	NN	O	O
of	NN	O	O
ATRA	NN	O	O
necessary	NN	O	O
to	NN	O	O
produce	NN	O	O
similar	NN	O	O
effects	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
AM580	NN	O	O
is	NN	O	O
not	NN	O	O
effective	NN	O	O
as	NN	O	O
ATRA	NN	O	O
in	NN	O	O
modulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
differentiation	NN	O	B-protein
markers	NN	O	I-protein
in	NN	O	O
the	NN	O	O
HL-60	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
patients	NN	O	O
during	NN	O	O
the	NN	O	O
stable	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

In	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
two	NN	O	O
other	NN	O	O
synthetic	NN	O	O
nonselective	NN	O	O
RAR	NN	O	O
ligands	NN	O	O
are	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
LAP	NN	O	B-protein
as	NN	O	O
much	NN	O	O
as	NN	O	O
AM580	NN	O	O
,	NN	O	O
whereas	NN	O	O
RAR	NN	O	O
beta-	NN	O	O
or	NN	O	O
RAR	NN	O	O
gamma-specific	NN	O	O
ligands	NN	O	O
are	NN	O	O
totally	NN	O	O
ineffective	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
AM580	NN	O	O
is	NN	O	O
more	NN	O	O
powerful	NN	O	O
than	NN	O	O
ATRA	NN	O	O
in	NN	O	O
modulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
differentiation	NN	O	O
antigens	NN	O	O
only	NN	O	O
in	NN	O	O
cells	NN	O	O
in	NN	O	O
which	NN	O	O
PML-RAR	NN	O	B-protein
is	NN	O	O
present	NN	O	O
.	NN	O	O

Binding	NN	O	O
experiments	NN	O	O
,	NN	O	O
using	NN	O	O
COS-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
transiently	NN	O	O
transfected	NN	O	O
with	NN	O	O
PML-RAR	NN	O	B-protein
and	NN	O	O
the	NN	O	O
normal	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
show	NN	O	O
that	NN	O	O
AM580	NN	O	O
has	NN	O	O
a	NN	O	O
lower	NN	O	O
affinity	NN	O	O
than	NN	O	O
ATRA	NN	O	O
for	NN	O	O
both	NN	O	O
receptors	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
PML-RAR	NN	O	B-protein
,	NN	O	O
the	NN	O	O
synthetic	NN	O	O
retinoid	NN	O	O
is	NN	O	O
a	NN	O	O
much	NN	O	O
better	NN	O	O
transactivator	NN	O	O
of	NN	O	O
retinoic	NN	O	B-DNA
acid-responsive	NN	O	I-DNA
element-containing	NN	O	I-DNA
promoters	NN	O	I-DNA
than	NN	O	O
the	NN	O	O
natural	NN	O	O
retinoid	NN	O	O
,	NN	O	O
whereas	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
AM580	NN	O	O
and	NN	O	O
ATRA	NN	O	O
have	NN	O	O
similar	NN	O	O
activity	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
explain	NN	O	O
the	NN	O	O
strong	NN	O	O
cyto-differentiating	NN	O	O
potential	NN	O	O
of	NN	O	O
AM580	NN	O	O
in	NN	O	O
PML-RAR-containing	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor-retinoid	NN	O	I-protein
X	NN	O	I-protein
receptor-vitamin	NN	O	I-protein
D	NN	O	I-protein
response	NN	O	I-protein
element	NN	O	I-protein
complex	NN	O	I-protein
formation	NN	O	O
by	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
vitamin	NN	O	B-cell_type
D-resistant	NN	O	I-cell_type
New	NN	O	I-cell_type
World	NN	O	I-cell_type
primate	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Most	NN	O	O
New	NN	O	O
World	NN	O	O
primate	NN	O	O
(	NN	O	O
NWP	NN	O	O
)	NN	O	O
genera	NN	O	O
evolved	NN	O	O
to	NN	O	O
require	NN	O	O
high	NN	O	O
circulating	NN	O	O
levels	NN	O	O
of	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
and	NN	O	O
vitamin	NN	O	O
D	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
an	NN	O	O
intracellular	NN	O	B-protein
vitamin	NN	O	I-protein
D	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
IDBP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
present	NN	O	O
in	NN	O	O
both	NN	O	O
nuclear	NN	O	O
and	NN	O	O
cytoplasmic	NN	O	O
fractions	NN	O	O
of	NN	O	O
NWP	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
or	NN	O	O
another	NN	O	O
protein	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
may	NN	O	O
cause	NN	O	O
or	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
steroid	NN	O	O
hormone-resistant	NN	O	O
state	NN	O	O
in	NN	O	O
NWP	NN	O	B-cell_line
by	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
dimerization	NN	O	O
process	NN	O	O
and/or	NN	O	O
by	NN	O	O
interference	NN	O	O
of	NN	O	O
receptor	NN	O	B-protein
complex	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
steroid-responsive	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
employed	NN	O	O
electromobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
to	NN	O	O
screen	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
proteins	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
vitamin	NN	O	B-DNA
D	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
VDRE	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
and	NN	O	O
post-nuclear	NN	O	O
extracts	NN	O	O
were	NN	O	O
prepared	NN	O	O
from	NN	O	O
two	NN	O	O
B-lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
known	NN	O	O
to	NN	O	O
be	NN	O	O
representative	NN	O	O
of	NN	O	O
the	NN	O	O
vitamin	NN	O	B-cell_line
D-resistant	NN	O	I-cell_line
and	NN	O	O
wild	NN	O	B-cell_line
type	NN	O	I-cell_line
phenotypes	NN	O	I-cell_line
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
extracts	NN	O	O
were	NN	O	O
compared	NN	O	O
for	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
retard	NN	O	O
the	NN	O	O
migration	NN	O	O
of	NN	O	O
radiolabeled	NN	O	O
double	NN	O	O
stranded	NN	O	O
oligomers	NN	O	O
representative	NN	O	O
of	NN	O	O
the	NN	O	O
VDREs	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
osteocalcin	NN	O	I-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
mouse	NN	O	I-DNA
osteopontin	NN	O	I-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
specific	NN	O	O
,	NN	O	O
retarded	NN	O	O
band	NN	O	O
containing	NN	O	O
VDR-RXR	NN	O	B-protein
was	NN	O	O
identified	NN	O	O
when	NN	O	O
wild	NN	O	O
type	NN	O	O
cell	NN	O	O
but	NN	O	O
not	NN	O	O
when	NN	O	O
vitamin	NN	O	O
D-resistant	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
was	NN	O	O
used	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
reaction	NN	O	O
with	NN	O	O
either	NN	O	O
probe	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
vitamin	NN	O	O
D-resistant	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
contained	NN	O	O
a	NN	O	O
protein	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
which	NN	O	O
was	NN	O	O
bound	NN	O	O
specifically	NN	O	O
to	NN	O	O
the	NN	O	O
VDRE	NN	O	B-DNA
and	NN	O	O
was	NN	O	O
capable	NN	O	O
of	NN	O	O
completely	NN	O	O
inhibiting	NN	O	O
VDR-RXR-VDRE	NN	O	B-protein
complex	NN	O	I-protein
formation	NN	O	O
;	NN	O	O
these	NN	O	O
effects	NN	O	O
were	NN	O	O
not	NN	O	O
demonstrated	NN	O	O
with	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
from	NN	O	O
the	NN	O	O
wild	NN	O	B-cell_line
type	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
or	NN	O	O
with	NN	O	O
the	NN	O	O
post-nuclear	NN	O	O
extract	NN	O	O
of	NN	O	O
the	NN	O	O
vitamin	NN	O	B-cell_line
D-resistant	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
a	NN	O	O
VDRE-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
,	NN	O	O
distinct	NN	O	O
from	NN	O	O
IDBP	NN	O	B-protein
and	NN	O	O
present	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
of	NN	O	O
cells	NN	O	O
from	NN	O	O
a	NN	O	O
prototypical	NN	O	B-cell_line
vitamin	NN	O	I-cell_line
D-resistant	NN	O	I-cell_line
NWP	NN	O	I-cell_line
,	NN	O	O
is	NN	O	O
capable	NN	O	O
of	NN	O	O
inhibiting	NN	O	O
normal	NN	O	O
VDR-RXR	NN	O	B-protein
heterodimer	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
VDRE	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

BCL-6	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
B-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Translocations	NN	O	O
involving	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-DNA
gene	NN	O	I-DNA
are	NN	O	O
common	NN	O	O
in	NN	O	O
the	NN	O	O
diffuse	NN	O	O
large	NN	O	O
cell	NN	O	O
subtype	NN	O	O
of	NN	O	O
non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
.	NN	O	O

Invariably	NN	O	O
,	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
coding	NN	O	O
region	NN	O	O
is	NN	O	O
intact	NN	O	O
,	NN	O	O
but	NN	O	O
its	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
untranslated	NN	O	I-DNA
region	NN	O	I-DNA
is	NN	O	O
replaced	NN	O	O
with	NN	O	O
sequences	NN	O	O
from	NN	O	O
the	NN	O	O
translocation	NN	O	B-DNA
partner	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
BCL-6	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
during	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Resting	NN	O	B-cell_type
B	NN	O	I-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
contain	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
mouse	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
anti-IgM	NN	O	B-protein
or	NN	O	I-protein
IgD	NN	O	I-protein
antibodies	NN	O	I-protein
,	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
,	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
plus	NN	O	O
ionomycin	NN	O	O
,	NN	O	O
or	NN	O	O
CD40	NN	O	B-protein
ligand	NN	O	I-protein
led	NN	O	O
to	NN	O	O
a	NN	O	O
five-fold	NN	O	O
to	NN	O	O
35-fold	NN	O	O
decrease	NN	O	O
in	NN	O	O
BCL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

Similar	NN	O	O
downregulation	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
seen	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
Staphylococcus	NN	O	O
aureus	NN	O	O
plus	NN	O	O
interleukin-2	NN	O	B-protein
or	NN	O	O
anti-IgM	NN	O	B-protein
antibodies	NN	O	I-protein
and	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
phytohemagglutinin	NN	O	B-protein
.	NN	O	O

BCL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
began	NN	O	O
to	NN	O	O
decrease	NN	O	O
8	NN	O	O
to	NN	O	O
16	NN	O	O
hours	NN	O	O
after	NN	O	O
stimulation	NN	O	O
,	NN	O	O
before	NN	O	O
cells	NN	O	O
entered	NN	O	O
S	NN	O	O
phase	NN	O	O
.	NN	O	O

Although	NN	O	O
polyclonal	NN	O	O
activation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
invariably	NN	O	O
decreased	NN	O	O
BCL-6	NN	O	B-protein
MRNA	NN	O	O
expression	NN	O	O
,	NN	O	O
activated	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
human	NN	O	O
germinal	NN	O	O
centers	NN	O	O
expressed	NN	O	O
BCL-6	NN	O	O
mRNA	NN	O	O
at	NN	O	O
levels	NN	O	O
comparable	NN	O	O
to	NN	O	O
the	NN	O	O
levels	NN	O	O
in	NN	O	O
resting	NN	O	O
B	NN	O	O
cells	NN	O	O
.	NN	O	O

Despite	NN	O	O
these	NN	O	O
similar	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
,	NN	O	O
BCL-6	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
was	NN	O	O
threefold	NN	O	O
to	NN	O	O
34-fold	NN	O	O
higher	NN	O	O
in	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
than	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
BCL-6	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
are	NN	O	O
controlled	NN	O	O
by	NN	O	O
translational	NN	O	O
or	NN	O	O
posttranslational	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
germinal	NN	O	O
center	NN	O	O
reaction	NN	O	O
provides	NN	O	O
unique	NN	O	O
activation	NN	O	O
signals	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
allow	NN	O	O
for	NN	O	O
continued	NN	O	O
,	NN	O	O
high-level	NN	O	O
BCL-6	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

DNA-binding	NN	O	B-protein
phosphoproteins	NN	O	I-protein
induced	NN	O	O
after	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
:	NN	O	O
effects	NN	O	O
of	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

To	NN	O	O
define	NN	O	O
novel	NN	O	B-protein
proteins	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
early	NN	O	O
transcriptional	NN	O	O
response	NN	O	O
during	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
used	NN	O	O
a	NN	O	O
high-resolution	NN	O	O
,	NN	O	O
two-dimensional	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
system	NN	O	O
to	NN	O	O
identify	NN	O	O
nuclear	NN	O	B-protein
,	NN	O	I-protein
deoxyribonucleic	NN	O	I-protein
acid	NN	O	I-protein
(	NN	O	I-protein
DNA	NN	O	I-protein
)	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
exhibiting	NN	O	O
rapid	NN	O	O
changes	NN	O	O
in	NN	O	O
phosphorylation	NN	O	O
following	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
18	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
whose	NN	O	O
phosphorylation	NN	O	O
level	NN	O	O
changed	NN	O	O
more	NN	O	O
than	NN	O	O
5-fold	NN	O	O
upon	NN	O	O
activation	NN	O	O
.	NN	O	O

Of	NN	O	O
these	NN	O	O
,	NN	O	O
11	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
possess	NN	O	O
DNA-binding	NN	O	O
properties	NN	O	O
.	NN	O	O

The	NN	O	O
11	NN	O	O
phosphoproteins	NN	O	B-protein
with	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
,	NN	O	O
along	NN	O	O
with	NN	O	O
4	NN	O	O
others	NN	O	O
,	NN	O	O
were	NN	O	O
analyzed	NN	O	O
further	NN	O	O
.	NN	O	O

Phosphoamino	NN	O	O
acid	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
several	NN	O	O
sets	NN	O	O
of	NN	O	O
proteins	NN	O	O
with	NN	O	O
different	NN	O	O
phosphorylated	NN	O	O
residues	NN	O	O
Kinetic	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
selected	NN	O	O
proteins	NN	O	O
was	NN	O	O
performed	NN	O	O
and	NN	O	O
revealed	NN	O	O
a	NN	O	O
complex	NN	O	O
group	NN	O	O
of	NN	O	O
transient	NN	O	O
and	NN	O	O
sustained	NN	O	O
responses	NN	O	O
to	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
activation-induced	NN	O	O
changes	NN	O	O
in	NN	O	O
one	NN	O	O
set	NN	O	O
of	NN	O	O
phosphoproteins	NN	O	B-protein
were	NN	O	O
dramatically	NN	O	O
inhibited	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
these	NN	O	O
phosphoproteins	NN	O	B-protein
may	NN	O	O
be	NN	O	O
directly	NN	O	O
involved	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
response	NN	O	O
to	NN	O	O
cellular	NN	O	O
activation	NN	O	O
by	NN	O	O
external	NN	O	O
stimuli	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inactivation	NN	O	O
of	NN	O	O
IkappaBbeta	NN	O	B-protein
by	NN	O	O
the	NN	O	O
tax	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
:	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
for	NN	O	O
constitutive	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

In	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
is	NN	O	O
sequestered	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
via	NN	O	O
interactions	NN	O	O
with	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
including	NN	O	O
IkappaBalpha	NN	O	B-protein
and	NN	O	O
IkappaBbeta	NN	O	B-protein
.	NN	O	O

During	NN	O	O
normal	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
IkappaBalpha	NN	O	B-protein
is	NN	O	O
rapidly	NN	O	O
phosphorylated	NN	O	O
,	NN	O	O
ubiquitinated	NN	O	O
,	NN	O	O
and	NN	O	O
degraded	NN	O	O
by	NN	O	O
the	NN	O	O
26S	NN	O	B-protein
proteasome	NN	O	I-protein
,	NN	O	O
thus	NN	O	O
permitting	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
functional	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
its	NN	O	O
transient	NN	O	O
pattern	NN	O	O
of	NN	O	O
nuclear	NN	O	O
induction	NN	O	O
during	NN	O	O
an	NN	O	O
immune	NN	O	O
response	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
is	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
in	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
transforming	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
HTLV-1	NN	O	O
Tax	NN	O	B-protein
targets	NN	O	O
IkappaBalpha	NN	O	B-protein
to	NN	O	O
the	NN	O	O
ubiquitin-proteasome	NN	O	O
pathway	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
remains	NN	O	O
unclear	NN	O	O
how	NN	O	O
this	NN	O	O
viral	NN	O	B-protein
protein	NN	O	I-protein
induces	NN	O	O
a	NN	O	O
persistent	NN	O	O
rather	NN	O	O
than	NN	O	O
transient	NN	O	O
NF-kappaB	NN	O	B-protein
response	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
acting	NN	O	O
on	NN	O	O
IkappaBalpha	NN	O	B-protein
,	NN	O	O
Tax	NN	O	B-protein
stimulates	NN	O	O
the	NN	O	O
turnover	NN	O	O
Of	NN	O	O
IkappaBbeta	NN	O	B-protein
via	NN	O	O
a	NN	O	O
related	NN	O	O
targeting	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Like	NN	O	O
IkappaBalpha	NN	O	B-protein
,	NN	O	O
Tax	NN	O	B-protein
-mediated	NN	O	O
breakdown	NN	O	O
of	NN	O	O
IkappaBbeta	NN	O	B-protein
in	NN	O	O
transfected	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
is	NN	O	O
blocked	NN	O	O
either	NN	O	O
by	NN	O	O
cell-permeable	NN	O	O
proteasome	NN	O	O
inhibitors	NN	O	O
or	NN	O	O
by	NN	O	O
mutation	NN	O	O
Of	NN	O	O
IkappaBbeta	NN	O	B-protein
at	NN	O	O
two	NN	O	O
serine	NN	O	O
residues	NN	O	O
present	NN	O	O
within	NN	O	O
its	NN	O	O
N-terminal	NN	O	B-protein
region	NN	O	I-protein
.	NN	O	O

Despite	NN	O	O
the	NN	O	O
dual	NN	O	O
specificity	NN	O	O
of	NN	O	O
HTLV-1	NN	O	O
Tax	NN	O	B-protein
for	NN	O	O
IkappaBalpha	NN	O	B-protein
and	NN	O	O
IkappaBbeta	NN	O	B-protein
at	NN	O	O
the	NN	O	O
protein	NN	O	O
level	NN	O	O
,	NN	O	O
Tax	NN	O	B-protein
selectively	NN	O	O
stimulates	NN	O	O
NF-kappaB	NN	O	B-protein
-directed	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IkappaBalpha	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
IkappaBbeta	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
is	NN	O	O
chronically	NN	O	O
downregulated	NN	O	O
in	NN	O	O
HTLV-1-infected	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
findings	NN	O	O
with	NN	O	O
IkappaBbeta	NN	O	B-protein
provide	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
Tax-expressing	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
p16	NN	O	B-protein
in	NN	O	O
the	NN	O	O
E2F-dependent	NN	O	B-protein
thymidine	NN	O	I-protein
kinase	NN	O	I-protein
regulation	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
alterations	NN	O	O
of	NN	O	O
the	NN	O	O
MTS1	NN	O	B-DNA
tumor	NN	O	I-DNA
suppressor	NN	O	I-DNA
gene	NN	O	I-DNA
on	NN	O	O
chromosome	NN	O	B-DNA
9p21	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
p16	NN	O	B-protein
,	NN	O	O
the	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
cyclin-dependent-kinase-4	NN	O	B-protein
and	NN	O	I-protein
6	NN	O	I-protein
,	NN	O	O
in	NN	O	O
tumorigenesis	NN	O	O
is	NN	O	O
not	NN	O	O
yet	NN	O	O
clear	NN	O	O
.	NN	O	O

Phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-protein
protein	NN	O	I-protein
by	NN	O	O
cyclin-dependent	NN	O	B-protein
kinases	NN	O	I-protein
4	NN	O	I-protein
and	NN	O	I-protein
6	NN	O	I-protein
prevents	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
E2F	NN	O	B-protein
,	NN	O	O
which	NN	O	O
subsequently	NN	O	O
promotes	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
S	NN	O	B-DNA
phase	NN	O	I-DNA
regulated	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
thymidine	NN	O	B-protein
kinase	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
a	NN	O	O
role	NN	O	O
of	NN	O	O
p16	NN	O	B-protein
in	NN	O	O
this	NN	O	O
regulation	NN	O	O
has	NN	O	O
been	NN	O	O
presumed	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
proof	NN	O	O
so	NN	O	O
far	NN	O	O
that	NN	O	O
loss	NN	O	O
of	NN	O	O
this	NN	O	O
tumor	NN	O	B-DNA
suppressor	NN	O	I-DNA
gene	NN	O	I-DNA
really	NN	O	O
affects	NN	O	O
E2F	NN	O	B-protein
-mediated	NN	O	O
regulations	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
thymidine	NN	O	B-protein
kinase	NN	O	I-protein
in	NN	O	O
phytohemagglutinin-stimulated	NN	O	B-cell_line
normal	NN	O	I-cell_line
human	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
the	NN	O	O
p16-negative	NN	O	B-cell_line
human	NN	O	I-cell_line
acute	NN	O	I-cell_line
lymphoblastic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
MOLT-4	NN	O	B-cell_line
and	NN	O	O
CEM	NN	O	B-cell_line
.	NN	O	O

Compared	NN	O	O
to	NN	O	O
normal	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
MOLT-4	NN	O	B-cell_line
and	NN	O	I-cell_line
CEM	NN	O	I-cell_line
cells	NN	O	I-cell_line
exhibited	NN	O	O
an	NN	O	O
altered	NN	O	O
cell	NN	O	O
cycle	NN	O	O
regulation	NN	O	O
of	NN	O	O
thymidine	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
a	NN	O	O
much	NN	O	O
higher	NN	O	O
intracellular	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
enzyme	NN	O	B-protein
,	NN	O	O
and	NN	O	O
higher	NN	O	O
thymidine	NN	O	B-RNA
kinase	NN	O	I-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
of	NN	O	O
p16	NN	O	B-protein
in	NN	O	O
normal	NN	O	O
human	NN	O	O
lymphocytes	NN	O	O
caused	NN	O	O
arrest	NN	O	O
in	NN	O	O
G1	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
without	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	O
growth	NN	O	O
of	NN	O	O
MOLT-4	NN	O	B-cell_line
and	NN	O	I-cell_line
CEM	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
all	NN	O	O
of	NN	O	O
them	NN	O	O
express	NN	O	O
functional	NN	O	O
retinoblastoma	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
leukemia	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
transient	NN	O	O
overexpression	NN	O	O
of	NN	O	O
p16	NN	O	B-protein
reestablished	NN	O	O
the	NN	O	O
normal	NN	O	O
regulation	NN	O	O
of	NN	O	O
thymidine	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
paralleled	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
of	NN	O	O
the	NN	O	O
underphosphorylated	NN	O	O
form	NN	O	O
of	NN	O	O
retinoblastoma	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
decrease	NN	O	O
of	NN	O	O
free	NN	O	O
E2F	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
its	NN	O	O
motif	NN	O	O
in	NN	O	O
the	NN	O	O
thymidine	NN	O	B-DNA
kinase	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
loss	NN	O	O
of	NN	O	O
p16	NN	O	B-protein
causes	NN	O	O
upregulation	NN	O	O
of	NN	O	O
this	NN	O	O
DNA	NN	O	O
precursor	NN	O	O
pathway	NN	O	O
enzyme	NN	O	B-protein
via	NN	O	O
activation	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
by	NN	O	O
a	NN	O	O
mechanism	NN	O	O
involving	NN	O	O
retinoblastoma	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Interactions	NN	O	O
of	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-DNA
activator	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
env	NN	O	B-DNA
gene	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
mammary	NN	O	O
tumor	NN	O	O
virus	NN	O	O
with	NN	O	O
activation-dependent	NN	O	B-protein
,	NN	O	I-protein
T	NN	O	I-protein
cell-specific	NN	O	I-protein
transacting	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
mouse	NN	O	O
mammary	NN	O	O
tumor	NN	O	O
virus	NN	O	O
env	NN	O	B-DNA
gene	NN	O	I-DNA
contains	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-DNA
activator	NN	O	I-DNA
(	NN	O	O
META	NN	O	B-DNA
)	NN	O	O
that	NN	O	O
can	NN	O	O
control	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
adjacent	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Transcriptional	NN	O	O
control	NN	O	O
by	NN	O	O
META	NN	O	B-DNA
parallels	NN	O	O
that	NN	O	O
of	NN	O	O
several	NN	O	O
lymphokine	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
being	NN	O	O
specific	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
dependent	NN	O	O
on	NN	O	O
their	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
drug	NN	O	O
cyclosporine	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
.	NN	O	O

DNase	NN	O	B-protein
I	NN	O	I-protein
footprinting	NN	O	O
indicated	NN	O	O
that	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
from	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
bound	NN	O	O
a	NN	O	O
promoter-proximal	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
META	NN	O	B-DNA
(	NN	O	I-DNA
P	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
promoter-distal	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
META	NN	O	B-DNA
(	NN	O	I-DNA
D+	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
within	NN	O	O
the	NN	O	O
400-base	NN	O	B-DNA
pair	NN	O	I-DNA
META	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Nuclear	NN	O	B-protein
factors	NN	O	I-protein
from	NN	O	O
unstimulated	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
from	NN	O	O
activated	NN	O	O
cells	NN	O	O
,	NN	O	O
bound	NN	O	O
a	NN	O	O
site	NN	O	O
,	NN	O	O
META	NN	O	B-DNA
(	NN	O	I-DNA
D-	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
adjacent	NN	O	O
to	NN	O	O
META	NN	O	B-DNA
(	NN	O	I-DNA
D+	NN	O	I-DNA
)	NN	O	I-DNA
.	NN	O	O

META	NN	O	B-DNA
(	NN	O	I-DNA
D+	NN	O	I-DNA
)	NN	O	I-DNA
directed	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
linked	NN	O	O
luciferase	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
gel	NN	O	O
shift	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
binding	NN	O	O
of	NN	O	O
inducible	NN	O	O
,	NN	O	O
CsA-sensitive	NN	O	B-protein
T	NN	O	I-protein
cell	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
in	NN	O	O
parallel	NN	O	O
with	NN	O	O
transfection	NN	O	O
results	NN	O	O
.	NN	O	O

Authentic	NN	O	O
NFAT	NN	O	B-DNA
and	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
targets	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
compete	NN	O	O
for	NN	O	O
the	NN	O	O
META	NN	O	B-protein
(	NN	O	I-protein
D+	NN	O	I-protein
)	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
SV40	NN	O	B-DNA
core	NN	O	I-DNA
sequence	NN	O	I-DNA
competed	NN	O	O
for	NN	O	O
META	NN	O	B-protein
(	NN	O	I-protein
D+	NN	O	I-protein
)	NN	O	I-protein
binding	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
but	NN	O	O
META	NN	O	B-DNA
(	NN	O	I-DNA
D+	NN	O	I-DNA
)	NN	O	I-DNA
failed	NN	O	O
to	NN	O	O
compete	NN	O	O
for	NN	O	O
the	NN	O	O
complexes	NN	O	O
obtained	NN	O	O
with	NN	O	O
the	NN	O	O
SV40	NN	O	O
probe	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
,	NN	O	O
taken	NN	O	O
together	NN	O	O
,	NN	O	O
indicate	NN	O	O
that	NN	O	O
META	NN	O	B-DNA
(	NN	O	I-DNA
D+	NN	O	I-DNA
)	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
novel	NN	O	O
transcriptional	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
similar	NN	O	O
in	NN	O	O
its	NN	O	O
cell-type	NN	O	O
specificity	NN	O	O
,	NN	O	O
activation	NN	O	O
dependence	NN	O	O
,	NN	O	O
and	NN	O	O
CsA	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
may	NN	O	O
be	NN	O	O
relevant	NN	O	O
to	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
MMTV	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
Mls	NN	O	B-protein
antigens	NN	O	I-protein
or	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
interferon	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
ICSAT/Pip/LSIRF	NN	O	B-protein
,	NN	O	O
that	NN	O	O
negatively	NN	O	O
regulates	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
interferon-regulated	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
isolated	NN	O	O
a	NN	O	O
novel	NN	O	B-DNA
cDNA	NN	O	I-DNA
clone	NN	O	I-DNA
encoding	NN	O	O
interferon	NN	O	B-protein
(	NN	O	I-protein
IFN	NN	O	I-protein
)	NN	O	I-protein
consensus	NN	O	I-protein
sequence-binding	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
adult	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
or	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
ICSAT	NN	O	B-protein
)	NN	O	O
;	NN	O	O
this	NN	O	O
protein	NN	O	O
is	NN	O	O
the	NN	O	O
human	NN	O	O
homolog	NN	O	O
of	NN	O	O
the	NN	O	O
recently	NN	O	O
cloned	NN	O	O
Pip/LSIRF	NN	O	B-protein
.	NN	O	O

ICSAT	NN	O	B-protein
is	NN	O	O
structurally	NN	O	O
most	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
previously	NN	O	O
cloned	NN	O	O
ICSBP	NN	O	B-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
IFN	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
IRF	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
proteins	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
interferon	NN	O	B-DNA
consensus	NN	O	I-DNA
sequences	NN	O	I-DNA
(	NN	O	O
ICSs	NN	O	B-DNA
)	NN	O	O
found	NN	O	O
in	NN	O	O
many	NN	O	O
promoters	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-regulated	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Among	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
investigated	NN	O	O
,	NN	O	O
ICSAT	NN	O	B-protein
was	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
virus	NN	O	I-cell_line
type	NN	O	I-cell_line
1	NN	O	I-cell_line
(	NN	O	I-cell_line
HTLV-1	NN	O	I-cell_line
)	NN	O	I-cell_line
-infected	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

When	NN	O	O
the	NN	O	O
HTLV-1	NN	O	B-DNA
tax	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
expressed	NN	O	O
or	NN	O	O
phorbol	NN	O	O
myristake	NN	O	O
acetate-A23187	NN	O	O
stimulation	NN	O	O
was	NN	O	O
used	NN	O	O
,	NN	O	O
ICSAT	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
induced	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
which	NN	O	O
otherwise	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
ICSAT	NN	O	B-protein
.	NN	O	O

When	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
ICSAT	NN	O	B-protein
to	NN	O	O
four	NN	O	O
different	NN	O	O
ICSs	NN	O	B-DNA
was	NN	O	O
tested	NN	O	O
,	NN	O	O
the	NN	O	O
relative	NN	O	O
differences	NN	O	O
in	NN	O	O
binding	NN	O	O
affinities	NN	O	O
for	NN	O	O
those	NN	O	O
ICSs	NN	O	B-DNA
were	NN	O	O
determined	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
functional	NN	O	O
role	NN	O	O
of	NN	O	O
ICSAT	NN	O	B-protein
,	NN	O	O
we	NN	O	O
performed	NN	O	O
cotransfection	NN	O	O
experiments	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
embryonal	NN	O	I-cell_line
carcinoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
N-Tera2	NN	O	I-cell_line
.	NN	O	O

ICSAT	NN	O	B-protein
was	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
possess	NN	O	O
repressive	NN	O	O
function	NN	O	O
over	NN	O	O
the	NN	O	O
gene	NN	O	O
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
IFN	NN	O	B-protein
stimulation	NN	O	O
or	NN	O	O
by	NN	O	O
IRF-1	NN	O	B-protein
cotransfection	NN	O	O
.	NN	O	O

Such	NN	O	O
repressive	NN	O	O
function	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
seen	NN	O	O
in	NN	O	O
IRF-2	NN	O	B-protein
or	NN	O	O
ICSBP	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
ICSAT	NN	O	B-protein
has	NN	O	O
a	NN	O	O
different	NN	O	O
repressive	NN	O	O
effect	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
IRF-2	NN	O	B-protein
or	NN	O	O
ICSBP	NN	O	B-protein
in	NN	O	O
some	NN	O	O
IFN-responsive	NN	O	B-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
novel	NN	O	O
mechanism	NN	O	O
of	NN	O	O
gene	NN	O	O
regulation	NN	O	O
by	NN	O	O
``	NN	O	O
differential	NN	O	O
repression	NN	O	O
''	NN	O	O
is	NN	O	O
used	NN	O	O
by	NN	O	O
multiple	NN	O	O
members	NN	O	O
of	NN	O	O
repressor	NN	O	B-protein
proteins	NN	O	I-protein
with	NN	O	O
different	NN	O	O
repressive	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
IFN-responsive	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
rise	NN	O	O
activated	NN	O	O
by	NN	O	O
doxepin-sensitive	NN	O	B-protein
H1-histamine	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
cloned	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
clarify	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
histamine	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
its	NN	O	O
transmembrane	NN	O	O
mechanism	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
agonists	NN	O	O
or	NN	O	O
antagonists	NN	O	O
of	NN	O	O
histamine	NN	O	B-protein
receptor	NN	O	I-protein
subtypes	NN	O	O
and	NN	O	O
bacterial	NN	O	O
toxins	NN	O	O
on	NN	O	O
intracellular	NN	O	O
concentration	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
)	NN	O	O
,	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
pyrilamine	NN	O	O
binding	NN	O	O
and	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
cloned	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

H1-agonists	NN	O	O
(	NN	O	O
histamine	NN	O	O
and	NN	O	O
2-methylhistamine	NN	O	O
)	NN	O	O
caused	NN	O	O
a	NN	O	O
transient	NN	O	O
rise	NN	O	O
of	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
,	NN	O	O
and	NN	O	O
H1-antagonists	NN	O	O
(	NN	O	O
pyrilamine	NN	O	O
and	NN	O	O
doxepin	NN	O	O
)	NN	O	O
inhibited	NN	O	O
the	NN	O	O
histamine-induced	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
rise	NN	O	O
more	NN	O	O
potently	NN	O	O
than	NN	O	O
the	NN	O	O
H2-antagonist	NN	O	O
(	NN	O	O
cimetidine	NN	O	O
)	NN	O	O
on	NN	O	O
the	NN	O	O
H3-antagonist	NN	O	O
(	NN	O	O
impromidine	NN	O	O
)	NN	O	O
.	NN	O	O

Binding	NN	O	O
parameters	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
pyrilamine	NN	O	O
binding	NN	O	O
were	NN	O	O
Kd	NN	O	O
=	NN	O	O
5.53	NN	O	O
nM	NN	O	O
and	NN	O	O
Bmax	NN	O	O
=	NN	O	O
2	NN	O	O
,	NN	O	O
647	NN	O	O
sites/cell	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
with	NN	O	O
B.pertussis	NN	O	O
,	NN	O	O
V.cholera.	NN	O	O
or	NN	O	O
C.botulinum	NN	O	O
toxin	NN	O	O
did	NN	O	O
not	NN	O	O
influence	NN	O	O
histamine-induced	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
rise	NN	O	O
.	NN	O	O

Western	NN	O	O
Blot	NN	O	O
analysis	NN	O	O
using	NN	O	O
antibodies	NN	O	O
against	NN	O	O
subunits	NN	O	O
of	NN	O	O
GTP-binding	NN	O	B-protein
proteins	NN	O	I-protein
indicated	NN	O	O
that	NN	O	O
Gq/G11	NN	O	B-protein
richly	NN	O	O
existed	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Histamine	NN	O	O
induced	NN	O	O
mRNA	NN	O	B-RNA
expression	NN	O	O
of	NN	O	O
an	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
gene	NN	O	I-DNA
c-fos	NN	O	B-DNA
.	NN	O	O

Pretreatment	NN	O	O
with	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
activator	NN	O	O
,	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
,	NN	O	O
caused	NN	O	O
almost	NN	O	O
complete	NN	O	O
inhibition	NN	O	O
of	NN	O	O
histamine-induced	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
rise	NN	O	O
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
do	NN	O	O
so	NN	O	O
by	NN	O	O
activators	NN	O	O
of	NN	O	O
cAMP-	NN	O	B-protein
and	NN	O	I-protein
cGMP-dependent	NN	O	I-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

STAT-related	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
patients	NN	O	O
.	NN	O	O

A	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
activated	NN	O	O
by	NN	O	O
many	NN	O	O
cytokines	NN	O	B-protein
has	NN	O	O
recently	NN	O	O
been	NN	O	O
elaborated	NN	O	O
.	NN	O	O

The	NN	O	O
JAK	NN	O	B-protein
kinases	NN	O	I-protein
and	NN	O	O
the	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
factors	NN	O	I-protein
have	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
essential	NN	O	O
components	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
STAT	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
oligonucleotide	NN	O	O
probes	NN	O	O
from	NN	O	O
the	NN	O	O
beta-casein	NN	O	B-DNA
and	NN	O	I-DNA
IRF-1	NN	O	I-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
ISRE	NN	O	O
probe	NN	O	O
to	NN	O	O
detect	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
acute	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
bandshift	NN	O	O
assays	NN	O	O
.	NN	O	O

Specific	NN	O	O
DNA	NN	O	O
protein	NN	O	B-protein
complex	NN	O	I-protein
formation	NN	O	O
was	NN	O	O
observed	NN	O	O
with	NN	O	O
the	NN	O	O
probes	NN	O	O
from	NN	O	O
the	NN	O	O
beta-casein	NN	O	O
and	NN	O	O
IRF-1	NN	O	B-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
the	NN	O	O
ISRE	NN	O	O
oligonucleotide	NN	O	O
probe	NN	O	O
,	NN	O	O
when	NN	O	O
cell	NN	O	O
extracts	NN	O	O
from	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ALL	NN	O	O
)	NN	O	O
and	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
(	NN	O	O
AML	NN	O	O
)	NN	O	O
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
nonradioactive	NN	O	O
oligonucleotides	NN	O	O
as	NN	O	O
competitors	NN	O	O
to	NN	O	O
show	NN	O	O
the	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
complex	NN	O	O
formation	NN	O	O
.	NN	O	O

Specific	NN	O	O
antibodies	NN	O	B-protein
directed	NN	O	O
against	NN	O	O
the	NN	O	O
individual	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
used	NN	O	O
in	NN	O	O
supershift	NN	O	O
experiments	NN	O	O
.	NN	O	O

STAT5-	NN	O	B-protein
and	NN	O	I-protein
STAT1-related	NN	O	I-protein
factors	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
in	NN	O	O
ALL	NN	O	O
and	NN	O	O
STAT1-	NN	O	B-protein
,	NN	O	I-protein
STAT3-	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
STAT5-related	NN	O	I-protein
proteins	NN	O	I-protein
were	NN	O	O
present	NN	O	O
in	NN	O	O
nuclear	NN	O	O
cell	NN	O	O
extracts	NN	O	O
from	NN	O	O
AML	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
not	NN	O	O
treated	NN	O	O
with	NN	O	O
cytokines	NN	O	B-protein
before	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
extracted	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
these	NN	O	O
factors	NN	O	O
are	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
likely	NN	O	O
that	NN	O	O
the	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
is	NN	O	O
a	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
events	NN	O	O
of	NN	O	O
leukemogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
element	NN	O	B-DNA
upstream	NN	O	O
from	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
delta-globin-encoding	NN	O	I-DNA
gene	NN	O	I-DNA
specifically	NN	O	O
enhances	NN	O	O
beta-globin	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
erythroleukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
factor	NN	O	I-protein
specific	NN	O	O
to	NN	O	O
adult	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
polypryrimidine-binding	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
PYBF	NN	O	B-protein
)	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
pyrimidine-rich	NN	O	B-DNA
region	NN	O	I-DNA
1	NN	O	B-DNA
kb	NN	O	I-DNA
upstream	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
delta-globin-encoding	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
HBD	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

The	NN	O	O
developmental	NN	O	O
stage-specificity	NN	O	O
of	NN	O	O
PYBF	NN	O	B-protein
and	NN	O	O
the	NN	O	O
location	NN	O	O
of	NN	O	O
its	NN	O	O
binding	NN	O	O
site	NN	O	O
between	NN	O	O
the	NN	O	O
fetal	NN	O	B-DNA
and	NN	O	I-DNA
adult	NN	O	I-DNA
beta-globin	NN	O	I-DNA
(	NN	O	I-DNA
HBB	NN	O	I-DNA
)	NN	O	I-DNA
-like	NN	O	I-DNA
genes	NN	O	I-DNA
suggest	NN	O	O
that	NN	O	O
PBYF	NN	O	B-protein
and	NN	O	O
its	NN	O	O
binding	NN	O	O
site	NN	O	O
may	NN	O	O
function	NN	O	O
in	NN	O	O
fetal-to-adult	NN	O	O
globin	NN	O	B-DNA
gene	NN	O	I-DNA
switching	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
383-bp	NN	O	B-DNA
(	NN	O	I-DNA
delta383	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	I-DNA
99-bp	NN	O	I-DNA
(	NN	O	I-DNA
delta99	NN	O	I-DNA
)	NN	O	I-DNA
sequences	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
PYBF-binding	NN	O	B-DNA
site	NN	O	I-DNA
on	NN	O	O
transcription	NN	O	O
from	NN	O	O
various	NN	O	O
globin	NN	O	O
and	NN	O	O
non-globin	NN	O	O
promoters	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
transient	NN	O	O
assay	NN	O	O
with	NN	O	O
the	NN	O	O
cat	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
murine	NN	O	B-cell_line
erythroleukemia	NN	O	I-cell_line
(	NN	O	I-cell_line
MEL	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
cell	NN	O	O
line	NN	O	O
with	NN	O	O
abundant	NN	O	O
PYBF	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
both	NN	O	O
delta383	NN	O	B-DNA
and	NN	O	O
delta99	NN	O	B-DNA
specifically	NN	O	O
enhance	NN	O	O
expression	NN	O	O
of	NN	O	O
cat	NN	O	B-protein
for	NN	O	O
plasmids	NN	O	B-DNA
containing	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
adult	NN	O	I-DNA
globin	NN	O	I-DNA
(	NN	O	I-DNA
HBB	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
whereas	NN	O	O
expression	NN	O	O
of	NN	O	O
similar	NN	O	O
constructs	NN	O	O
using	NN	O	O
human	NN	O	B-DNA
fetal	NN	O	I-DNA
(	NN	O	I-DNA
A	NN	O	I-DNA
gamma-	NN	O	I-DNA
)	NN	O	I-DNA
globin	NN	O	I-DNA
(	NN	O	I-DNA
HBG1	NN	O	I-DNA
)	NN	O	I-DNA
or	NN	O	I-DNA
simian	NN	O	I-DNA
virus	NN	O	I-DNA
40	NN	O	I-DNA
(	NN	O	I-DNA
SV40	NN	O	I-DNA
)	NN	O	I-DNA
promoters	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
enhanced	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
PYBF	NN	O	B-protein
and	NN	O	O
the	NN	O	O
pyrimidine-rich	NN	O	B-DNA
region	NN	O	I-DNA
upstream	NN	O	O
from	NN	O	O
HBD	NN	O	B-DNA
can	NN	O	O
specifically	NN	O	O
enhance	NN	O	O
HBB	NN	O	B-DNA
transcription	NN	O	O
in	NN	O	O
adult	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Translocation	NN	O	B-DNA
breakpoints	NN	O	I-DNA
in	NN	O	O
three	NN	O	O
patients	NN	O	O
with	NN	O	O
campomelic	NN	O	O
dysplasia	NN	O	O
and	NN	O	O
autosomal	NN	O	O
sex	NN	O	O
reversal	NN	O	O
map	NN	O	O
more	NN	O	O
than	NN	O	O
130	NN	O	O
kb	NN	O	O
from	NN	O	O
SOX9	NN	O	B-DNA
.	NN	O	O

Campomelic	NN	O	O
dysplasia	NN	O	O
(	NN	O	O
CMPD1	NN	O	O
)	NN	O	O
and	NN	O	O
autosomal	NN	O	O
XY	NN	O	O
sex	NN	O	O
reversal	NN	O	O
(	NN	O	O
SRA1	NN	O	O
)	NN	O	O
are	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
SRY-related	NN	O	B-DNA
gene	NN	O	I-DNA
SOX9	NN	O	B-DNA
on	NN	O	O
17q	NN	O	B-DNA
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
the	NN	O	O
17q	NN	O	B-DNA
breakpoints	NN	O	I-DNA
in	NN	O	O
four	NN	O	O
CMPD1	NN	O	O
translocation	NN	O	O
cases	NN	O	O
previously	NN	O	O
analyzed	NN	O	O
by	NN	O	O
us	NN	O	O
and	NN	O	O
others	NN	O	O
map	NN	O	O
50	NN	O	O
kb	NN	O	O
or	NN	O	O
more	NN	O	O
from	NN	O	O
SOX9	NN	O	B-DNA
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
present	NN	O	O
clinical	NN	O	O
,	NN	O	O
cytogenetic	NN	O	O
,	NN	O	O
and	NN	O	O
molecular	NN	O	O
data	NN	O	O
from	NN	O	O
a	NN	O	O
new	NN	O	O
CMPD1/SRA1	NN	O	O
patient	NN	O	O
with	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
6	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q14	NN	O	I-DNA
;	NN	O	I-DNA
q24	NN	O	I-DNA
)	NN	O	I-DNA
.	NN	O	O

Fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
has	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
17q	NN	O	B-DNA
breakpoint	NN	O	I-DNA
in	NN	O	O
this	NN	O	O
case	NN	O	O
maps	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
region	NN	O	O
as	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
in	NN	O	O
the	NN	O	O
other	NN	O	O
translocation	NN	O	O
cases	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
130	NN	O	O
kb	NN	O	O
from	NN	O	O
SOX9	NN	O	B-DNA
.	NN	O	O

Likewise	NN	O	O
,	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
in	NN	O	O
two	NN	O	O
of	NN	O	O
the	NN	O	O
previously	NN	O	O
described	NN	O	O
cases	NN	O	O
also	NN	O	O
map	NN	O	O
more	NN	O	O
than	NN	O	O
130	NN	O	O
kb	NN	O	O
and	NN	O	O
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
pulsed	NN	O	O
field	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
analysis	NN	O	O
,	NN	O	O
at	NN	O	O
most	NN	O	O
400	NN	O	O
kb	NN	O	O
or	NN	O	O
690	NN	O	O
kb	NN	O	O
from	NN	O	O
SOX9	NN	O	B-DNA
.	NN	O	O

By	NN	O	O
using	NN	O	O
a	NN	O	O
SOX9	NN	O	B-DNA
coding	NN	O	I-DNA
sequence	NN	O	I-DNA
polymorphism	NN	O	I-DNA
,	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
SOX9	NN	O	B-DNA
alleles	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
the	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
in	NN	O	O
lymphoblastoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
translocation	NN	O	O
cases	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
isoform	NN	O	O
of	NN	O	O
the	NN	O	O
Spi-B	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

Spi-B	NN	O	B-protein
is	NN	O	O
an	NN	O	O
Ets	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
related	NN	O	O
to	NN	O	O
the	NN	O	O
oncoprotein	NN	O	B-protein
Spi-1/PU.1	NN	O	B-protein
and	NN	O	O
highly	NN	O	O
expressed	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
Ets	NN	O	B-protein
proteins	NN	O	I-protein
share	NN	O	O
a	NN	O	O
conserved	NN	O	O
Ets	NN	O	B-protein
domain	NN	O	I-protein
that	NN	O	O
mediates	NN	O	O
specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

Spi-B	NN	O	B-protein
binds	NN	O	O
DNA	NN	O	B-DNA
sequences	NN	O	I-DNA
containing	NN	O	O
a	NN	O	O
core	NN	O	O
5'-GGAA-3	NN	O	O
'	NN	O	O
and	NN	O	O
activates	NN	O	O
transcription	NN	O	O
through	NN	O	O
this	NN	O	O
motif	NN	O	O
.	NN	O	O

Up	NN	O	O
to	NN	O	O
date	NN	O	O
,	NN	O	O
the	NN	O	O
biological	NN	O	O
function	NN	O	O
of	NN	O	O
Spi-B	NN	O	B-protein
remains	NN	O	O
unknown	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
an	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
variant	NN	O	O
of	NN	O	O
Spi-B	NN	O	B-protein
,	NN	O	O
named	NN	O	O
deltaSpi-B	NN	O	B-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
lost	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

In	NN	O	O
B	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
deltaspi-B	NN	O	B-RNA
and	NN	O	I-RNA
spi-B	NN	O	I-RNA
mRNAs	NN	O	I-RNA
were	NN	O	O
present	NN	O	O
simultaneously	NN	O	O
in	NN	O	O
a	NN	O	O
ratio	NN	O	O
of	NN	O	O
around	NN	O	O
10	NN	O	O
%	NN	O	O
.	NN	O	O

DeltaSpi-B	NN	O	B-protein
product	NN	O	O
was	NN	O	O
not	NN	O	O
able	NN	O	O
to	NN	O	O
bind	NN	O	O
DNA	NN	O	O
and	NN	O	O
was	NN	O	O
recovered	NN	O	O
in	NN	O	O
cytoplasmic	NN	O	O
cellular	NN	O	O
extracts	NN	O	O
.	NN	O	O

We	NN	O	O
raise	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
delta	NN	O	B-protein
Spi-B	NN	O	I-protein
might	NN	O	O
affect	NN	O	O
Spi-B	NN	O	B-protein
function	NN	O	O
by	NN	O	O
recruiting	NN	O	O
factors	NN	O	O
involved	NN	O	O
in	NN	O	O
Spi-B	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Binding	NN	O	O
and	NN	O	O
cooperative	NN	O	O
interactions	NN	O	O
between	NN	O	O
two	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
transcriptional	NN	O	I-protein
coactivators	NN	O	I-protein
.	NN	O	O

The	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
and	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
octamer-binding	NN	O	I-protein
protein	NN	O	I-protein
1/octamer-binding	NN	O	I-protein
factor	NN	O	I-protein
1/Oct	NN	O	I-protein
coactivator	NN	O	I-protein
from	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
Bob1/OBF-1/OCA-B	NN	O	B-protein
)	NN	O	O
represent	NN	O	O
two	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
transcriptional	NN	O	I-protein
coactivators	NN	O	I-protein
.	NN	O	O

CIITA	NN	O	B-protein
and	NN	O	O
Bob1	NN	O	B-protein
interact	NN	O	O
with	NN	O	O
proteins	NN	O	O
that	NN	O	O
bind	NN	O	O
to	NN	O	O
conserved	NN	O	B-DNA
upstream	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
major	NN	O	I-DNA
histocompatibility	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
octamer-binding	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Both	NN	O	O
CIITA	NN	O	B-protein
and	NN	O	O
Bob1	NN	O	B-protein
increase	NN	O	O
the	NN	O	O
expression	NN	O	O
from	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
prototypic	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
,	NN	O	O
interactions	NN	O	O
between	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
and	NN	O	O
Bob1	NN	O	B-protein
are	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
octamer-binding	NN	O	B-RNA
site	NN	O	I-RNA
.	NN	O	O

Using	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
binding	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
confirm	NN	O	O
that	NN	O	O
Bob1	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
CIITA	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
CIITA	NN	O	B-protein
not	NN	O	O
only	NN	O	O
activates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
but	NN	O	O
recruits	NN	O	O
another	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
coactivator	NN	O	O
to	NN	O	O
increase	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Efficient	NN	O	O
transcription	NN	O	O
and	NN	O	O
replication	NN	O	O
of	NN	O	O
simian	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-DNA
and	NN	O	I-DNA
Sp1	NN	O	I-DNA
binding	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Ten	NN	O	O
mutants	NN	O	O
of	NN	O	O
the	NN	O	O
simian	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
SIV	NN	O	O
)	NN	O	O
SIVmac239	NN	O	O
bearing	NN	O	O
deletions	NN	O	O
(	NN	O	O
delta	NN	O	O
)	NN	O	O
or	NN	O	O
substitutions	NN	O	O
(	NN	O	O
subst	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-DNA
and/or	NN	O	O
Sp1	NN	O	B-DNA
binding	NN	O	I-DNA
elements	NN	O	I-DNA
were	NN	O	O
created	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
replicative	NN	O	O
capacities	NN	O	O
of	NN	O	O
the	NN	O	O
mutants	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

All	NN	O	O
mutants	NN	O	O
,	NN	O	O
including	NN	O	O
one	NN	O	O
extensively	NN	O	O
mutagenized	NN	O	O
strain	NN	O	O
entirely	NN	O	O
missing	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-DNA
and	NN	O	I-DNA
four	NN	O	I-DNA
Spl	NN	O	I-DNA
binding	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
replicated	NN	O	O
with	NN	O	O
wild-type	NN	O	O
kinetics	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
wild-type	NN	O	O
level	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_line
blood	NN	O	I-cell_line
mononuclear	NN	O	I-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
in	NN	O	O
50	NN	O	O
to	NN	O	O
100	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
experiments	NN	O	O
.	NN	O	O

One	NN	O	O
group	NN	O	O
of	NN	O	O
mutants	NN	O	O
replicated	NN	O	O
very	NN	O	O
similarly	NN	O	O
to	NN	O	O
SIVmac239	NN	O	O
in	NN	O	O
kinetics	NN	O	O
and	NN	O	O
yield	NN	O	O
in	NN	O	O
CEMxl74	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
2x	NN	O	O
NFKappaB	NN	O	B-protein
>	NN	O	O
or	NN	O	O
=	NN	O	O
SlVmac239	NN	O	B-protein
approximately	NN	O	O
deltaNFkappaB	NN	O	B-protein
approximately	NN	O	O
deltaSpl234	NN	O	B-protein
approximately	NN	O	O
substNFkappaB	NN	O	B-protein
approximately	NN	O	O
substSpl2	NN	O	B-protein
approximately	NN	O	O
substSp23	NN	O	B-protein
)	NN	O	O
,	NN	O	O
while	NN	O	O
a	NN	O	O
second	NN	O	O
group	NN	O	O
replicated	NN	O	O
with	NN	O	O
delayed	NN	O	O
or	NN	O	O
slightly	NN	O	O
delayed	NN	O	O
kinetics	NN	O	O
in	NN	O	O
CEMxl74	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
SIVmac239	NN	O	O
>	NN	O	O
substSp34	NN	O	B-protein
>	NN	O	O
deltaNFkappaBdeltaSpl234	NN	O	B-protein
approximately	NN	O	O
deltaNFkappaBdeltaSp1	NN	O	B-protein
>	NN	O	O
substSpl234	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Reversions	NN	O	O
or	NN	O	O
additional	NN	O	O
mutations	NN	O	O
were	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
U3	NN	O	B-DNA
and	NN	O	I-DNA
R	NN	O	I-DNA
regions	NN	O	I-DNA
of	NN	O	O
proviral	NN	O	B-DNA
DNA	NN	O	I-DNA
from	NN	O	O
CEMxl74	NN	O	B-cell_line
cells	NN	O	I-cell_line
infected	NN	O	O
with	NN	O	O
the	NN	O	O
SIVmac239	NN	O	O
mutants	NN	O	O
.	NN	O	O

Similar	NN	O	O
results	NN	O	O
were	NN	O	O
obtained	NN	O	O
when	NN	O	O
mutants	NN	O	O
of	NN	O	O
SIVmacMER	NN	O	O
(	NN	O	O
a	NN	O	O
macrophage-competent	NN	O	O
derivative	NN	O	O
of	NN	O	O
SIVmac239	NN	O	O
)	NN	O	O
were	NN	O	O
tested	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cell	NN	O	I-cell_type
and	NN	O	O
CEMx174	NN	O	B-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
most	NN	O	O
mutated	NN	O	O
viruses	NN	O	O
was	NN	O	O
suppressed	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
rhesus	NN	O	I-cell_type
monkey	NN	O	I-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
(	NN	O	O
SIVmacMER	NN	O	O
approximately	NN	O	O
2xNFkappaB	NN	O	B-protein
approximately	NN	O	O
substNFkappaB	NN	O	B-protein
>	NN	O	O
deltaNFkappaB	NN	O	B-protein
>	NN	O	O
deltaNFkappaB	NN	O	B-protein
deltaSpl234	NN	O	B-protein
approximately	NN	O	O
deltaNFkappaBdeltaSpl	NN	O	B-protein
>	NN	O	O
deltaSpl234	NN	O	B-protein
approximately	NN	O	O
substSpl2	NN	O	B-protein
>	NN	O	O
substSp23	NN	O	B-protein
approximately	NN	O	O
substSp34	NN	O	B-protein
approximately	NN	O	O
substSpl234	NN	O	B-protein
>	NN	O	O
or	NN	O	O
=	NN	O	O
SIVmac239	NN	O	O
)	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
Sp1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
had	NN	O	O
the	NN	O	O
most	NN	O	O
dramatic	NN	O	O
effects	NN	O	O
on	NN	O	O
SIVmac	NN	O	O
replication	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_line
macrophage	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
-driven	NN	O	O
secreted	NN	O	O
alkaline	NN	O	B-protein
phosphatase	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
transient	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
,	NN	O	O
unlike	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
,	NN	O	O
the	NN	O	O
SIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
possesses	NN	O	O
an	NN	O	O
enhancer	NN	O	B-DNA
region	NN	O	I-DNA
just	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-DNA
element	NN	O	I-DNA
which	NN	O	O
maintains	NN	O	O
significant	NN	O	O
levels	NN	O	O
of	NN	O	O
basal	NN	O	O
transcription	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-DNA
and	NN	O	I-DNA
Sp1	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
region	NN	O	O
is	NN	O	O
responsive	NN	O	O
to	NN	O	O
transactivation	NN	O	O
by	NN	O	O
Tat	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
SIV	NN	O	B-DNA
TATA	NN	O	I-DNA
box	NN	O	I-DNA
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
stronger	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
surprisingly	NN	O	O
high	NN	O	O
replicative	NN	O	O
capacity	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	O
and	NN	O	O
Sp1	NN	O	O
binding	NN	O	O
site	NN	O	O
mutants	NN	O	O
of	NN	O	O
SIVmac	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
unique	NN	O	O
features	NN	O	O
or	NN	O	O
the	NN	O	O
enhancer/promoter	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

cAMP	NN	O	O
inducibility	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
repressor	NN	O	I-protein
ICER	NN	O	B-protein
in	NN	O	O
developing	NN	O	O
and	NN	O	O
mature	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
Proc	NN	O	O
Natl	NN	O	O
Acad	NN	O	O
Sci	NN	O	O
U	NN	O	O
S	NN	O	O
A	NN	O	O
1996	NN	O	O
Jul	NN	O	O
23	NN	O	O
;	NN	O	O
93	NN	O	O
(	NN	O	O
15	NN	O	O
)	NN	O	O
:	NN	O	O
8154	NN	O	O
]	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
cAMP-dependent	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
exerts	NN	O	O
an	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and	NN	O	O
effector	NN	O	O
functions	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
form	NN	O	O
high	NN	O	O
intracellular	NN	O	O
levels	NN	O	O
of	NN	O	O
cAMP	NN	O	O
is	NN	O	O
acquired	NN	O	O
during	NN	O	O
development	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
thymus	NN	O	O
and	NN	O	O
is	NN	O	O
retained	NN	O	O
by	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
mature	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
elevated	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
potent	NN	O	B-protein
transcriptional	NN	O	I-protein
repressor	NN	O	I-protein
ICER	NN	O	B-protein
(	NN	O	O
inducible	NN	O	B-protein
cAMP	NN	O	I-protein
early	NN	O	I-protein
repressor	NN	O	I-protein
)	NN	O	O
previously	NN	O	O
described	NN	O	O
in	NN	O	O
the	NN	O	O
hypothalamic-pituitary-gonadal	NN	O	O
axis	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
assays	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
ICER	NN	O	B-protein
inhibits	NN	O	O
calcineurin	NN	O	B-protein
-mediated	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
promoter	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
Tax	NN	O	B-protein
-mediated	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
T-lymphotropic	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
I	NN	O	I-DNA
(	NN	O	I-DNA
HTLV-I	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
ICER	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
cAMP-induced	NN	O	O
quiescence	NN	O	O
and	NN	O	O
the	NN	O	O
persistent	NN	O	O
latency	NN	O	O
of	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
hydrophobic	NN	O	O
domain	NN	O	O
of	NN	O	O
Ca2+-modulating	NN	O	B-protein
cyclophilin	NN	O	I-protein
ligand	NN	O	I-protein
modulates	NN	O	O
calcium	NN	O	O
influx	NN	O	O
signaling	NN	O	O
in	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

Ca2+-modulating	NN	O	B-protein
cyclophilin	NN	O	I-protein
ligand	NN	O	I-protein
(	NN	O	O
CAML	NN	O	B-protein
)	NN	O	O
was	NN	O	O
originally	NN	O	O
described	NN	O	O
as	NN	O	O
a	NN	O	O
cyclophilin	NN	O	B-protein
B-binding	NN	O	I-protein
protein	NN	O	I-protein
whose	NN	O	O
overexpression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
causes	NN	O	O
a	NN	O	O
rise	NN	O	O
in	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
,	NN	O	O
thus	NN	O	O
activating	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
early	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

As	NN	O	O
reported	NN	O	O
here	NN	O	O
,	NN	O	O
structure-function	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
CAML	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
indicates	NN	O	O
that	NN	O	O
two	NN	O	O
of	NN	O	O
CAML	NN	O	B-protein
's	NN	O	O
putative	NN	O	B-protein
membrane-spanning	NN	O	I-protein
domains	NN	O	I-protein
are	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
hydrophobic	NN	O	B-protein
C-terminal	NN	O	I-protein
tail	NN	O	I-protein
of	NN	O	O
CAML	NN	O	B-protein
forms	NN	O	O
its	NN	O	O
effector	NN	O	O
domain	NN	O	O
,	NN	O	O
thus	NN	O	O
implicating	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
hydrophilic	NN	O	I-protein
domain	NN	O	I-protein
in	NN	O	O
a	NN	O	O
regulatory	NN	O	O
role	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
define	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
protein	NN	O	I-protein
motif	NN	O	I-protein
that	NN	O	O
functions	NN	O	O
in	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
signaling	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus-encoded	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
EBNA-5	NN	O	I-protein
accumulates	NN	O	O
in	NN	O	O
PML-containing	NN	O	O
bodies	NN	O	O
.	NN	O	O

EBNA-5	NN	O	B-protein
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
(	NN	O	I-protein
EBV	NN	O	I-protein
)	NN	O	I-protein
-encoded	NN	O	I-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
required	NN	O	O
for	NN	O	O
immortalization	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
EBV-transformed	NN	O	B-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
EBNA-5	NN	O	B-protein
is	NN	O	O
preferentially	NN	O	O
targetted	NN	O	O
to	NN	O	O
distinct	NN	O	O
nuclear	NN	O	O
foci	NN	O	O
.	NN	O	O

Previously	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
(	NN	O	O
W.Q.	NN	O	O
Jiang	NN	O	O
,	NN	O	O
L.Szekely	NN	O	O
,	NN	O	O
V.Wendel-Hansen	NN	O	O
,	NN	O	O
N.Ringertz	NN	O	O
,	NN	O	O
G.Klein	NN	O	O
,	NN	O	O
and	NN	O	O
A.	NN	O	O
Rosen	NN	O	O
,	NN	O	O
Exp.Cell	NN	O	O
Res	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
new	NN	O	O
variant	NN	O	O
translocation	NN	O	O
in	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukaemia	NN	O	O
:	NN	O	O
molecular	NN	O	O
characterization	NN	O	O
and	NN	O	O
clinical	NN	O	O
correlation	NN	O	O
.	NN	O	O

Translocation	NN	O	B-DNA
t	NN	O	I-DNA
(	NN	O	I-DNA
15	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q22	NN	O	I-DNA
;	NN	O	I-DNA
q21	NN	O	I-DNA
)	NN	O	O
is	NN	O	O
an	NN	O	O
acquired	NN	O	O
clonal	NN	O	O
cytogenetic	NN	O	O
change	NN	O	O
present	NN	O	O
in	NN	O	O
almost	NN	O	O
all	NN	O	O
cases	NN	O	O
of	NN	O	O
acute	NN	O	O
promelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
genetic	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
translocation	NN	O	O
supports	NN	O	O
its	NN	O	O
integral	NN	O	O
role	NN	O	O
in	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
APL	NN	O	O
in	NN	O	O
whom	NN	O	O
the	NN	O	O
leukaemic	NN	O	O
clone	NN	O	O
was	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
true	NN	O	O
variant	NN	O	O
of	NN	O	O
the	NN	O	O
classical	NN	O	B-DNA
t	NN	O	I-DNA
(	NN	O	I-DNA
15	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
patient	NN	O	O
whose	NN	O	O
disease	NN	O	O
had	NN	O	O
numerous	NN	O	O
atypical	NN	O	O
clinical	NN	O	O
features	NN	O	O
,	NN	O	O
had	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
11	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q13	NN	O	I-DNA
;	NN	O	I-DNA
121	NN	O	I-DNA
)	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
chromosome	NN	O	B-DNA
17	NN	O	I-DNA
breakpoint	NN	O	I-DNA
was	NN	O	O
localized	NN	O	O
to	NN	O	O
intron	NN	O	B-DNA
2	NN	O	I-DNA
of	NN	O	O
RARA	NN	O	B-DNA
by	NN	O	O
Southern	NN	O	O
blotting	NN	O	O
,	NN	O	O
and	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
evidence	NN	O	O
at	NN	O	O
the	NN	O	O
molecular	NN	O	O
level	NN	O	O
for	NN	O	O
rearrangement	NN	O	O
at	NN	O	O
PML	NN	O	B-DNA
locus	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
,	NN	O	O
along	NN	O	O
with	NN	O	O
previous	NN	O	O
reports	NN	O	O
of	NN	O	O
rare	NN	O	O
variant	NN	O	O
translocations	NN	O	O
in	NN	O	O
APL	NN	O	O
,	NN	O	O
indicate	NN	O	O
that	NN	O	O
while	NN	O	O
dysregulation	NN	O	O
of	NN	O	O
RARA	NN	O	B-DNA
by	NN	O	O
gene	NN	O	O
fusion	NN	O	O
may	NN	O	O
be	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
APL	NN	O	O
phenotype	NN	O	O
,	NN	O	O
the	NN	O	O
particular	NN	O	O
fusion	NN	O	O
partner	NN	O	O
may	NN	O	O
determine	NN	O	O
clinicopathological	NN	O	O
aspects	NN	O	O
,	NN	O	O
including	NN	O	O
presentation	NN	O	O
,	NN	O	O
response	NN	O	O
to	NN	O	O
treatment	NN	O	O
with	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
prognosis	NN	O	O
.	NN	O	O

This	NN	O	O
heterogeneity	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
variant	NN	O	O
fusion	NN	O	B-protein
partners	NN	O	I-protein
of	NN	O	O
RARA	NN	O	B-DNA
in	NN	O	O
APL	NN	O	O
encode	NN	O	O
factors	NN	O	B-protein
with	NN	O	O
properties	NN	O	O
both	NN	O	O
common	NN	O	O
to	NN	O	O
and	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
PML	NN	O	B-cell_type
.	NN	O	O

Investigation	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
promises	NN	O	O
to	NN	O	O
shed	NN	O	O
light	NN	O	O
on	NN	O	O
the	NN	O	O
complex	NN	O	O
development	NN	O	O
pathways	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
haematopoiesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
model	NN	O	O
of	NN	O	O
latent	NN	O	O
adenovirus	NN	O	O
5	NN	O	O
infection	NN	O	O
in	NN	O	O
the	NN	O	O
guinea	NN	O	O
pig	NN	O	O
(	NN	O	O
Cavia	NN	O	O
porcellus	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
model	NN	O	O
of	NN	O	O
adenovirus	NN	O	O
5	NN	O	O
(	NN	O	O
Ad5	NN	O	O
)	NN	O	O
infection	NN	O	O
was	NN	O	O
developed	NN	O	O
in	NN	O	O
guinea	NN	O	O
pigs	NN	O	O
to	NN	O	O
begin	NN	O	O
to	NN	O	O
study	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
peripheral	NN	O	O
lung	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Forty	NN	O	O
animals	NN	O	O
were	NN	O	O
inoculated	NN	O	O
intranasally	NN	O	O
with	NN	O	O
10	NN	O	O
(	NN	O	O
7.0	NN	O	O
)	NN	O	O
pfu	NN	O	O
of	NN	O	O
Ad5/animal	NN	O	O
,	NN	O	O
and	NN	O	O
15	NN	O	O
animals	NN	O	O
inoculated	NN	O	O
with	NN	O	O
sterile	NN	O	O
culture	NN	O	O
media	NN	O	O
served	NN	O	O
as	NN	O	O
controls	NN	O	O
.	NN	O	O

Viral	NN	O	O
titres	NN	O	O
were	NN	O	O
10	NN	O	O
(	NN	O	O
4.4	NN	O	O
)	NN	O	O
,	NN	O	O
10	NN	O	O
(	NN	O	O
6.1	NN	O	O
)	NN	O	O
,	NN	O	O
10	NN	O	O
(	NN	O	O
5.2	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
10	NN	O	O
(	NN	O	O
2.9	NN	O	O
)	NN	O	O
pfu/animal	NN	O	O
,	NN	O	O
on	NN	O	O
days	NN	O	O
1	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
and	NN	O	O
7	NN	O	O
after	NN	O	O
infection	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

In	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
to	NN	O	O
viral	NN	O	B-DNA
DNA	NN	O	I-DNA
and	NN	O	O
immunocytochemistry	NN	O	O
for	NN	O	O
Ad5	NN	O	O
E1A	NN	O	B-protein
protein	NN	O	I-protein
localized	NN	O	O
the	NN	O	O
virus	NN	O	O
to	NN	O	O
airway	NN	O	O
and	NN	O	O
alveolar	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Histologic	NN	O	O
examination	NN	O	O
showed	NN	O	O
an	NN	O	O
extensive	NN	O	O
inflammatory	NN	O	O
cell	NN	O	O
infiltration	NN	O	O
around	NN	O	O
the	NN	O	O
airways	NN	O	O
,	NN	O	O
with	NN	O	O
epithelial	NN	O	O
necrosis	NN	O	O
and	NN	O	O
an	NN	O	O
alveolar	NN	O	O
exudate	NN	O	O
that	NN	O	O
caused	NN	O	O
localized	NN	O	O
alveolar	NN	O	O
collapse	NN	O	O
in	NN	O	O
the	NN	O	O
infected	NN	O	O
areas	NN	O	O
.	NN	O	O

Immunocytochemistry	NN	O	O
identified	NN	O	O
the	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
infiltrate	NN	O	O
as	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
all	NN	O	O
animals	NN	O	O
20	NN	O	O
and	NN	O	O
47	NN	O	O
days	NN	O	O
after	NN	O	O
infection	NN	O	O
had	NN	O	O
seroconverted	NN	O	O
to	NN	O	O
Ad5	NN	O	O
,	NN	O	O
virus	NN	O	O
was	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
these	NN	O	O
groups	NN	O	O
either	NN	O	O
by	NN	O	O
viral	NN	O	O
plaque	NN	O	O
assay	NN	O	O
or	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

Ad5	NN	O	B-DNA
E1A	NN	O	I-DNA
DNA	NN	O	I-DNA
was	NN	O	O
detected	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
in	NN	O	O
five	NN	O	O
of	NN	O	O
six	NN	O	O
animals	NN	O	O
20	NN	O	O
days	NN	O	O
after	NN	O	O
infection	NN	O	O
and	NN	O	O
in	NN	O	O
five	NN	O	O
of	NN	O	O
five	NN	O	O
animals	NN	O	O
47	NN	O	O
days	NN	O	O
after	NN	O	O
infection	NN	O	O
.	NN	O	O

In	NN	O	O
these	NN	O	O
same	NN	O	O
animals	NN	O	O
,	NN	O	O
E1A	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
20	NN	O	O
days	NN	O	O
after	NN	O	O
infection	NN	O	O
in	NN	O	O
two	NN	O	O
and	NN	O	O
47	NN	O	O
days	NN	O	O
after	NN	O	O
infection	NN	O	O
in	NN	O	O
one	NN	O	O
while	NN	O	O
persistent	NN	O	O
bronchiolitis	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
four	NN	O	O
and	NN	O	O
three	NN	O	O
animals	NN	O	O
20	NN	O	O
and	NN	O	O
47	NN	O	O
days	NN	O	O
after	NN	O	O
infection	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
guinea	NN	O	O
pig	NN	O	O
provides	NN	O	O
a	NN	O	O
useful	NN	O	O
model	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Ad5	NN	O	O
infection	NN	O	O
in	NN	O	O
chronic	NN	O	O
airway	NN	O	O
inflammation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cyclic	NN	O	O
AMP	NN	O	O
-responsive	NN	O	B-DNA
element	NN	O	I-DNA
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
zebra	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
6	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
shown	NN	O	O
that	NN	O	O
infection	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus	NN	O	I-cell_line
(	NN	O	I-cell_line
EBV	NN	O	I-cell_line
)	NN	O	I-cell_line
genome-positive	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
6	NN	O	O
(	NN	O	O
HHV-6	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
immediate-early	NN	O	B-DNA
EBV	NN	O	I-DNA
Zebra	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
followed	NN	O	O
by	NN	O	O
virus	NN	O	O
replication	NN	O	O
(	NN	O	O
L.Flamand	NN	O	O
,	NN	O	O
I.Stefanescu	NN	O	O
,	NN	O	O
D.V.Ablashi	NN	O	O
,	NN	O	O
and	NN	O	O
J.Menezes	NN	O	O
,	NN	O	O
J.Virol.67	NN	O	O
:	NN	O	O
6768-6777	NN	O	O
,	NN	O	O
1993	NN	O	O
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
HHV-6	NN	O	O
upregulates	NN	O	O
Zebra	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
through	NN	O	O
a	NN	O	O
cyclic	NN	O	B-DNA
AMP-responsive	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CRE	NN	O	B-DNA
)	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
Zebra	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
Zp	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Using	NN	O	O
human	NN	O	B-cell_line
B-	NN	O	I-cell_line
or	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
transfected	NN	O	O
with	NN	O	O
ZpCat	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
region	NN	O	O
designated	NN	O	O
the	NN	O	O
ZII	NN	O	B-DNA
domain	NN	O	I-DNA
of	NN	O	O
Zp	NN	O	B-DNA
is	NN	O	O
the	NN	O	O
target	NN	O	O
of	NN	O	O
HHV-6	NN	O	O
transactivation	NN	O	O
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
AP-1/CRE	NN	O	I-DNA
site	NN	O	I-DNA
within	NN	O	O
ZII	NN	O	B-DNA
abolished	NN	O	O
the	NN	O	O
inducibility	NN	O	O
of	NN	O	O
Zp	NN	O	B-DNA
by	NN	O	O
HHV-6	NN	O	O
,	NN	O	O
whereas	NN	O	O
positioning	NN	O	O
of	NN	O	O
the	NN	O	O
ZII	NN	O	B-DNA
domain	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
minimal	NN	O	I-DNA
promoter	NN	O	I-DNA
conferred	NN	O	O
responsiveness	NN	O	O
following	NN	O	O
HHV-6	NN	O	O
infection	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
to	NN	O	O
ZII	NN	O	B-DNA
was	NN	O	O
prevented	NN	O	O
by	NN	O	O
oligonucleotides	NN	O	O
containing	NN	O	O
CRE	NN	O	B-DNA
but	NN	O	O
not	NN	O	O
by	NN	O	O
AP-1	NN	O	B-DNA
consensus	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

Antibodies	NN	O	O
against	NN	O	O
CRE-binding	NN	O	B-protein
(	NN	O	I-protein
CREB	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
but	NN	O	O
not	NN	O	O
against	NN	O	O
c-Fos	NN	O	B-protein
or	NN	O	O
c-Jun	NN	O	B-protein
were	NN	O	O
able	NN	O	O
to	NN	O	O
supershift	NN	O	O
the	NN	O	O
DNA-protein	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
identifying	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
which	NN	O	O
binds	NN	O	O
to	NN	O	O
ZII	NN	O	B-DNA
as	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
CREB	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
transfection	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
protein	NN	O	B-DNA
kinase	NN	O	I-DNA
A	NN	O	I-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
were	NN	O	O
found	NN	O	O
to	NN	O	O
activate	NN	O	O
Zp	NN	O	B-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
phosphorylated	NN	O	O
form	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
protein	NN	O	I-protein
can	NN	O	O
play	NN	O	O
a	NN	O	O
determining	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
EBV	NN	O	O
reactivation	NN	O	O
process	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
in	NN	O	O
stably	NN	O	O
transfected	NN	O	O
Jurkat	NN	O	B-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

Two	NN	O	O
Jurkat	NN	O	B-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
have	NN	O	O
been	NN	O	O
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
vector	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Upon	NN	O	O
stimulation	NN	O	O
,	NN	O	O
they	NN	O	O
express	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
secreted	NN	O	O
heat	NN	O	O
stable	NN	O	O
placental	NN	O	B-protein
alkaline	NN	O	I-protein
phosphatase	NN	O	I-protein
.	NN	O	O

With	NN	O	O
these	NN	O	O
clones	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
both	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
(	NN	O	O
IC50	NN	O	O
=	NN	O	O
8	NN	O	O
nM	NN	O	O
)	NN	O	O
and	NN	O	O
FK506	NN	O	O
(	NN	O	O
IC50	NN	O	O
=	NN	O	O
160	NN	O	O
pM	NN	O	O
)	NN	O	O
,	NN	O	O
presumably	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Selective	NN	O	O
phosphatase	NN	O	O
inhibitors	NN	O	O
for	NN	O	O
protein	NN	O	B-protein
phosphatase	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
PP1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
2A	NN	O	B-protein
(	NN	O	O
PP2A	NN	O	B-protein
)	NN	O	O
that	NN	O	O
do	NN	O	O
not	NN	O	O
inhibit	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
and	NN	O	O
calyculin	NN	O	O
A	NN	O	O
,	NN	O	O
also	NN	O	O
inhibited	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
with	NN	O	O
IC50s	NN	O	O
of	NN	O	O
87	NN	O	O
nM	NN	O	O
and	NN	O	O
4	NN	O	O
nM	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
and	NN	O	O
related	NN	O	O
inhibitors	NN	O	O
may	NN	O	O
block	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
through	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
PP1	NN	O	B-protein
,	NN	O	O
instead	NN	O	O
of	NN	O	O
PP2A	NN	O	B-protein
.	NN	O	O

NFAT	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
also	NN	O	O
inhibited	NN	O	O
by	NN	O	O
agents	NN	O	O
that	NN	O	O
increase	NN	O	O
cAMP	NN	O	O
concentrations	NN	O	O
such	NN	O	O
as	NN	O	O
dibutyryl	NN	O	O
cAMP	NN	O	O
,	NN	O	O
forskolin	NN	O	O
and	NN	O	O
prostaglandin	NN	O	O
E2	NN	O	O
.	NN	O	O

These	NN	O	O
stable	NN	O	O
Jurkat	NN	O	B-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
provide	NN	O	O
a	NN	O	O
convenient	NN	O	O
and	NN	O	O
sensitive	NN	O	O
tool	NN	O	O
to	NN	O	O
study	NN	O	O
NFAT	NN	O	B-protein
regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoids	NN	O	O
induced	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
mRNA	NN	O	O
expression	NN	O	O
in	NN	O	O
asthma	NN	O	O
.	NN	O	O

Although	NN	O	O
their	NN	O	O
precise	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
,	NN	O	O
glucocorticoids	NN	O	O
represent	NN	O	O
the	NN	O	O
most	NN	O	O
effective	NN	O	O
therapy	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
asthma	NN	O	O
.	NN	O	O

Interactions	NN	O	O
between	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
complex	NN	O	I-protein
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
regulate	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
some	NN	O	O
genes	NN	O	O
,	NN	O	O
including	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
itself	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
compare	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
seven	NN	O	O
unstable	NN	O	O
untreated	NN	O	O
asthmatic	NN	O	O
patients	NN	O	O
who	NN	O	O
were	NN	O	O
subsequently	NN	O	O
treated	NN	O	O
with	NN	O	O
high	NN	O	O
doses	NN	O	O
of	NN	O	O
parenteral	NN	O	O
corticosteroid	NN	O	O
(	NN	O	O
methyl	NN	O	O
prednisolone	NN	O	O
120	NN	O	O
mg/day	NN	O	O
)	NN	O	O
for	NN	O	O
10	NN	O	O
days	NN	O	O
.	NN	O	O

mRNA	NN	O	B-RNA
expression	NN	O	O
was	NN	O	O
identified	NN	O	O
after	NN	O	O
RNA	NN	O	O
extraction	NN	O	O
using	NN	O	O
RNAzol	NN	O	O
and	NN	O	O
analysed	NN	O	O
after	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
,	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
using	NN	O	O
a	NN	O	O
semiquantitative	NN	O	O
competitive	NN	O	O
hybridization	NN	O	O
assay	NN	O	O
.	NN	O	O

All	NN	O	O
asthmatic	NN	O	O
patients	NN	O	O
showed	NN	O	O
an	NN	O	O
improvement	NN	O	O
in	NN	O	O
their	NN	O	O
FEV1	NN	O	O
values	NN	O	O
after	NN	O	O
corticosteroid	NN	O	O
treatment	NN	O	O
(	NN	O	O
per	NN	O	O
cent	NN	O	O
of	NN	O	O
predicted	NN	O	O
value	NN	O	O
68.28	NN	O	O
+/-	NN	O	O
4.93	NN	O	O
versus	NN	O	O
95.57	NN	O	O
+/-	NN	O	O
6.41	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.02	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
mRNA	NN	O	B-RNA
expression	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.02	NN	O	O
)	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
their	NN	O	O
monocytes	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
report	NN	O	O
of	NN	O	O
an	NN	O	O
ex	NN	O	O
vivo	NN	O	O
down-regulation	NN	O	O
for	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
mRNA	NN	O	B-RNA
expression	NN	O	O
,	NN	O	O
following	NN	O	O
corticosteroid	NN	O	O
treatment	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mapping	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
repression	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
lymphoid-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
oct-2A	NN	O	B-protein
.	NN	O	O

The	NN	O	O
lymphoid-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Oct-2a	NN	O	B-protein
is	NN	O	O
implicated	NN	O	O
in	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
via	NN	O	O
the	NN	O	O
octamer	NN	O	O
motif	NN	O	O
.	NN	O	O

Structure/function	NN	O	O
analysis	NN	O	O
of	NN	O	O
various	NN	O	O
Oct-2a	NN	O	B-protein
effector	NN	O	I-protein
regions	NN	O	I-protein
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
GAL4	NN	O	B-protein
DNA-binding	NN	O	I-protein
domain	NN	O	I-protein
revealed	NN	O	O
that	NN	O	O
Oct-2a	NN	O	B-protein
contains	NN	O	O
two	NN	O	O
functionally	NN	O	O
different	NN	O	O
activation	NN	O	B-protein
domains	NN	O	I-protein
at	NN	O	O
the	NN	O	O
N	NN	O	B-protein
and	NN	O	I-protein
the	NN	O	I-protein
C	NN	O	I-protein
termini	NN	O	I-protein
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
both	NN	O	O
domains	NN	O	O
is	NN	O	O
strongly	NN	O	O
potentiated	NN	O	O
by	NN	O	O
interactions	NN	O	O
with	NN	O	O
distinct	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
coactivators	NN	O	I-protein
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
repression	NN	O	B-protein
domain	NN	O	I-protein
located	NN	O	O
within	NN	O	O
the	NN	O	O
N	NN	O	B-protein
terminus	NN	O	I-protein
of	NN	O	O
Oct-2a	NN	O	B-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
2-99	NN	O	I-protein
)	NN	O	O
.	NN	O	O

When	NN	O	O
this	NN	O	O
domain	NN	O	O
was	NN	O	O
transferred	NN	O	O
to	NN	O	O
a	NN	O	O
potent	NN	O	O
activator	NN	O	O
,	NN	O	O
transcription	NN	O	O
was	NN	O	O
strongly	NN	O	O
inhibited	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
present	NN	O	O
a	NN	O	O
deletion	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
Oct-2a	NN	O	B-protein
to	NN	O	O
determine	NN	O	O
the	NN	O	O
minimal	NN	O	B-protein
repression	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
stretch	NN	O	O
of	NN	O	O
23	NN	O	O
amino	NN	O	O
acids	NN	O	O
,	NN	O	O
rich	NN	O	O
in	NN	O	O
serine	NN	O	O
and	NN	O	O
threonine	NN	O	O
residues	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
responsible	NN	O	O
for	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
repression	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
repression	NN	O	O
is	NN	O	O
strongly	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
type	NN	O	O
of	NN	O	O
enhancer	NN	O	B-DNA
present	NN	O	O
in	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
plasmid	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
tested	NN	O	O
.	NN	O	O

The	NN	O	O
possibility	NN	O	O
that	NN	O	O
Oct-2a	NN	O	B-protein
can	NN	O	O
act	NN	O	O
as	NN	O	O
an	NN	O	O
activator	NN	O	O
and/or	NN	O	O
a	NN	O	O
repressor	NN	O	O
may	NN	O	O
have	NN	O	O
important	NN	O	O
consequences	NN	O	O
for	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
Oct-2a	NN	O	B-protein
in	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
other	NN	O	O
developmental	NN	O	O
processes	NN	O	O
.	NN	O	O

-DOCSTART-	O

Opposing	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
on	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
neutrophilic	NN	O	B-cell_type
and	NN	O	I-cell_type
eosinophilic	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
.	NN	O	O

Eosinophils	NN	O	B-cell_type
and	NN	O	O
neutrophils	NN	O	B-cell_type
are	NN	O	O
closely	NN	O	O
related	NN	O	O
,	NN	O	O
terminally	NN	O	B-cell_type
differentiated	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
in	NN	O	O
vitro	NN	O	O
undergo	NN	O	O
constitutive	NN	O	O
cell	NN	O	O
death	NN	O	O
by	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
onset	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
can	NN	O	O
be	NN	O	O
delayed	NN	O	O
by	NN	O	O
hemopoietins	NN	O	B-protein
and	NN	O	O
inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
there	NN	O	O
have	NN	O	O
been	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
reports	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
(	NN	O	O
in	NN	O	O
particular	NN	O	O
dexamethasone	NN	O	O
)	NN	O	O
antagonize	NN	O	O
the	NN	O	O
eosinophil	NN	O	O
life-prolonging	NN	O	O
effects	NN	O	O
of	NN	O	O
hemopoietins	NN	O	B-protein
,	NN	O	O
direct	NN	O	O
effects	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
on	NN	O	O
eosinophil	NN	O	O
apoptosis	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
documented	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
direct	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
on	NN	O	O
eosinophil	NN	O	B-cell_type
and	NN	O	O
neutrophil	NN	O	B-cell_type
apoptosis	NN	O	O
in	NN	O	O
light	NN	O	O
of	NN	O	O
their	NN	O	O
common	NN	O	O
therapeutic	NN	O	O
use	NN	O	O
as	NN	O	O
anti-inflammatory	NN	O	O
and	NN	O	O
anti-allergic/hypereosinophilic	NN	O	O
agents	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
treatment	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
induced	NN	O	O
eosinophil	NN	O	B-cell_type
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
neutrophil	NN	O	B-cell_type
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
on	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
was	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
abolished	NN	O	O
by	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
antagonist	NN	O	O
RU38486	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
description	NN	O	O
of	NN	O	O
an	NN	O	O
agent	NN	O	O
that	NN	O	O
promotes	NN	O	O
eosinophil	NN	O	B-cell_type
apoptosis	NN	O	O
while	NN	O	O
inhibiting	NN	O	O
neutrophil	NN	O	B-cell_type
apoptosis	NN	O	O
,	NN	O	O
and	NN	O	O
thus	NN	O	O
presents	NN	O	O
a	NN	O	O
novel	NN	O	O
approach	NN	O	O
to	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
control	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
these	NN	O	O
closely	NN	O	O
related	NN	O	O
cell	NN	O	O
types	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
increases	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
clinical	NN	O	O
action	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
inflammation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Permanent	NN	O	O
occupancy	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
enhancer	NN	O	I-DNA
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
needed	NN	O	O
for	NN	O	O
persistent	NN	O	O
viral	NN	O	O
replication	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

This	NN	O	O
work	NN	O	O
aimed	NN	O	O
to	NN	O	O
ascertain	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
kappaB-responsive	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	I-DNA
HIV-1	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
not	NN	O	O
only	NN	O	O
in	NN	O	O
early	NN	O	O
initiation	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
long-term	NN	O	O
maintenance	NN	O	O
of	NN	O	O
proviral	NN	O	O
transcription	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

For	NN	O	O
this	NN	O	O
purpose	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
three	NN	O	O
main	NN	O	O
approaches	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
was	NN	O	O
to	NN	O	O
abruptly	NN	O	O
terminate	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor-induced	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
enhancer	NN	O	O
sequences	NN	O	O
in	NN	O	O
U1	NN	O	O
monocytic	NN	O	O
cells	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
short	NN	O	O
pulse	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

This	NN	O	O
resulted	NN	O	O
in	NN	O	O
concomitant	NN	O	O
decrease	NN	O	O
in	NN	O	O
nuclear	NN	O	O
NF-kappaB	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
and	NN	O	O
endogenous	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
was	NN	O	O
to	NN	O	O
suppress	NN	O	O
the	NN	O	O
permanent	NN	O	O
NF-kappaB	NN	O	B-protein
translocation	NN	O	O
induced	NN	O	O
by	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
itself	NN	O	O
in	NN	O	O
chronically	NN	O	B-cell_line
infected	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
using	NN	O	O
a	NN	O	O
specific	NN	O	O
proteasome	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
Z-LLL-H	NN	O	O
)	NN	O	O
.	NN	O	O

As	NN	O	O
early	NN	O	O
as	NN	O	O
2	NN	O	O
h	NN	O	O
after	NN	O	O
addition	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitor	NN	O	O
to	NN	O	O
the	NN	O	O
culture	NN	O	O
medium	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
an	NN	O	O
inhibition	NN	O	O
of	NN	O	O
both	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
HIV-1	NN	O	O
genome	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
third	NN	O	O
approach	NN	O	O
was	NN	O	O
to	NN	O	O
monitor	NN	O	O
the	NN	O	O
replication	NN	O	O
competence	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
an	NN	O	O
infectious	NN	O	O
HIV-1	NN	O	O
provirus	NN	O	O
carrying	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
kappaB-responsive	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
both	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeats	NN	O	I-DNA
.	NN	O	O

Compared	NN	O	O
with	NN	O	O
its	NN	O	O
wild-type	NN	O	O
counterpart	NN	O	O
,	NN	O	O
this	NN	O	O
mutated	NN	O	O
provirus	NN	O	O
showed	NN	O	O
a	NN	O	O
profoundly	NN	O	O
decreased	NN	O	O
,	NN	O	O
Z-LLL-H-insensitive	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
replicative	NN	O	O
activity	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
.	NN	O	O

Together	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
occupancy	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
enhancer	NN	O	I-DNA
by	NN	O	O
NF-kappaB	NN	O	B-protein
(	NN	O	I-protein
p50/p65	NN	O	I-protein
)	NN	O	I-protein
heterodimers	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
ongoing	NN	O	O
transcription	NN	O	O
of	NN	O	O
integrated	NN	O	O
HIV	NN	O	O
provirus	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
even	NN	O	O
in	NN	O	O
cells	NN	O	O
chronically	NN	O	O
infected	NN	O	O
and	NN	O	O
permanently	NN	O	O
producing	NN	O	O
functional	NN	O	O
HIV	NN	O	B-protein
Tat	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappaB	NN	O	B-protein
is	NN	O	O
crucial	NN	O	O
to	NN	O	O
the	NN	O	O
intense	NN	O	O
self-perpetuated	NN	O	O
viral	NN	O	O
transcription	NN	O	O
observed	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Rel-deficient	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
exhibit	NN	O	O
defects	NN	O	O
in	NN	O	O
production	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
3	NN	O	I-protein
and	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
c-rel	NN	O	B-DNA
protooncogene	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
subunit	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B-like	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Mice	NN	O	O
lacking	NN	O	O
Rel	NN	O	B-protein
are	NN	O	O
defective	NN	O	O
in	NN	O	O
mitogenic	NN	O	O
activation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
display	NN	O	O
impaired	NN	O	O
humoral	NN	O	O
immunity	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
attempt	NN	O	O
to	NN	O	O
identify	NN	O	O
changes	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
that	NN	O	O
accompany	NN	O	O
the	NN	O	O
T-cell	NN	O	O
stimulation	NN	O	O
defects	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
Rel	NN	O	B-protein
,	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
activation	NN	O	I-protein
markers	NN	O	I-protein
and	NN	O	O
cytokine	NN	O	O
production	NN	O	O
in	NN	O	O
mitogen-stimulated	NN	O	O
Rel-/-	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
cell	NN	O	O
surface	NN	O	O
markers	NN	O	O
including	NN	O	O
the	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	I-protein
IL-2R	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
CD25	NN	O	B-protein
)	NN	O	O
,	NN	O	O
CD69	NN	O	B-protein
and	NN	O	O
L-selectin	NN	O	B-protein
(	NN	O	O
CD62	NN	O	B-protein
)	NN	O	O
is	NN	O	O
normal	NN	O	O
in	NN	O	O
mitogen-activated	NN	O	B-cell_line
Rel-/-	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
is	NN	O	O
impaired	NN	O	O
.	NN	O	O

In	NN	O	O
Rel-/-	NN	O	B-cell_line
splenic	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
stimulated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
,	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-3	NN	O	O
,	NN	O	O
IL-5	NN	O	O
,	NN	O	O
granulocyte-	NN	O	B-protein
macrophage	NN	O	I-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
were	NN	O	O
only	NN	O	O
2-	NN	O	O
to	NN	O	O
3-fold	NN	O	O
lower	NN	O	O
compared	NN	O	O
with	NN	O	O
normal	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
anti-CD3	NN	O	B-protein
and	NN	O	O
anti-CD28	NN	O	B-protein
stimulated	NN	O	O
Rel-/-	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
fail	NN	O	O
to	NN	O	O
proliferate	NN	O	O
,	NN	O	O
make	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
detectable	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

Exogenous	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
which	NN	O	O
restitutes	NN	O	O
the	NN	O	O
proliferative	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
anti-CD3-	NN	O	B-cell_line
and	NN	O	I-cell_line
anti-CD28-treated	NN	O	I-cell_line
Rel-/-	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
restores	NN	O	O
production	NN	O	O
of	NN	O	O
IL-5	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-3	NN	O	B-protein
and	NN	O	O
GM-CSF	NN	O	B-protein
expression	NN	O	O
to	NN	O	O
approximately	NN	O	O
normal	NN	O	O
levels	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
mitogen-activated	NN	O	B-cell_line
Rel-/-	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
lipopolysaccharide-stimulated	NN	O	O
Rel-/-	NN	O	B-cell_line
macrophages	NN	O	I-cell_line
produce	NN	O	O
higher	NN	O	O
than	NN	O	O
normal	NN	O	O
levels	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
establish	NN	O	O
that	NN	O	O
Rel	NN	O	B-protein
can	NN	O	O
function	NN	O	O
as	NN	O	O
an	NN	O	O
activator	NN	O	O
or	NN	O	O
repressor	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
is	NN	O	O
required	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
for	NN	O	O
production	NN	O	O
of	NN	O	O
IL-3	NN	O	B-protein
and	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	O
appearance	NN	O	O
of	NN	O	O
a	NN	O	O
factor	NN	O	O
that	NN	O	O
binds	NN	O	O
the	NN	O	O
CD28	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
correlates	NN	O	O
with	NN	O	O
interleukin-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
requires	NN	O	O
the	NN	O	O
combined	NN	O	O
signaling	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
costimulatory	NN	O	B-protein
molecules	NN	O	I-protein
such	NN	O	O
as	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
produce	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
control	NN	O	O
point	NN	O	O
in	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
IL-2	NN	O	B-protein
enhancer	NN	O	O
contains	NN	O	O
a	NN	O	O
functional	NN	O	O
motif	NN	O	O
named	NN	O	O
CD28	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CD28RE	NN	O	B-DNA
)	NN	O	O
that	NN	O	O
serves	NN	O	O
a	NN	O	O
role	NN	O	O
as	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
mitogenic	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
.	NN	O	O

The	NN	O	O
CD28RE	NN	O	B-DNA
sequence	NN	O	O
reveals	NN	O	O
similarity	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
kappaB	NN	O	I-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
CD28RE	NN	O	B-DNA
binds	NN	O	O
an	NN	O	O
inducible	NN	O	O
protein	NN	O	O
with	NN	O	O
a	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
approximately	NN	O	O
35	NN	O	O
kDa	NN	O	O
called	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
mitogenic-activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-MATp35	NN	O	B-protein
)	NN	O	O
that	NN	O	O
is	NN	O	O
clearly	NN	O	O
different	NN	O	O
from	NN	O	O
the	NN	O	O
known	NN	O	O
NF-	NN	O	B-protein
kappaB/Rel	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
NF-MATp35	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
depend	NN	O	O
on	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
and	NN	O	O
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
received	NN	O	O
a	NN	O	O
mitogenic	NN	O	O
combination	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
stimuli	NN	O	O
,	NN	O	O
not	NN	O	O
necessarily	NN	O	O
including	NN	O	O
CD28	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Nonmitogenic	NN	O	O
T	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
did	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
appearance	NN	O	O
of	NN	O	O
NF-MATp35	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
mitogenic	NN	O	O
combinations	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
are	NN	O	O
integrated	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
NF-MATp35	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
its	NN	O	O
effect	NN	O	O
on	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
inhibited	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-MATp35	NN	O	B-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
nuclear	NN	O	O
appearance	NN	O	O
of	NN	O	O
NF-MATp35	NN	O	B-protein
shows	NN	O	O
excellent	NN	O	O
correlation	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
unique	NN	O	O
characteristic	NN	O	O
among	NN	O	O
nuclear	NN	O	O
factors	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
basis	NN	O	O
for	NN	O	O
hyporesponsiveness	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
inducible	NN	O	I-DNA
nitric	NN	O	I-DNA
oxide	NN	O	I-DNA
synthase	NN	O	I-DNA
gene	NN	O	I-DNA
to	NN	O	O
lipopolysaccharide/interferon-gamma	NN	O	O
.	NN	O	O

The	NN	O	O
work	NN	O	O
reported	NN	O	O
here	NN	O	O
resolves	NN	O	O
,	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
gene	NN	O	O
regulation	NN	O	O
,	NN	O	O
the	NN	O	O
controversy	NN	O	O
as	NN	O	O
to	NN	O	O
whether	NN	O	O
or	NN	O	O
not	NN	O	O
human	NN	O	B-cell_type
monocytes/macrophages	NN	O	I-cell_type
can	NN	O	O
produce	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
(	NN	O	O
NO	NN	O	O
)	NN	O	O
when	NN	O	O
stimulated	NN	O	O
with	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
co-stimulation	NN	O	O
by	NN	O	O
interferon-gamma	NN	O	O
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Studies	NN	O	O
included	NN	O	O
structural	NN	O	O
comparison	NN	O	O
of	NN	O	O
the	NN	O	O
promoters	NN	O	O
for	NN	O	O
human	NN	O	O
and	NN	O	O
mouse	NN	O	O
inducible	NN	O	B-DNA
NO	NN	O	I-DNA
synthase	NN	O	I-DNA
(	NN	O	I-DNA
iNOS	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
transfection	NN	O	O
and	NN	O	O
assay	NN	O	O
of	NN	O	O
human	NN	O	O
and	NN	O	O
mouse	NN	O	O
iNOS	NN	O	B-DNA
promoter	NN	O	I-DNA
regions	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
+/-	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
of	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Two	NN	O	O
explanations	NN	O	O
for	NN	O	O
hyporesponsiveness	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
iNOS	NN	O	I-DNA
promoter	NN	O	I-DNA
to	NN	O	O
LPS	NN	O	O
+/-	NN	O	O
IFN-gamma	NN	O	B-protein
were	NN	O	O
found	NN	O	O
:	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
multiple	NN	O	O
inactivating	NN	O	O
nucleotide	NN	O	O
substitutions	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
counterpart	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
that	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
regulate	NN	O	O
LPS/	NN	O	O
IFN-gamma	NN	O	B-protein
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
iNOS	NN	O	I-DNA
gene	NN	O	I-DNA
;	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
and	NN	O	O
absence	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
(	NN	O	O
e.g.	NN	O	O
,	NN	O	O
an	NN	O	O
LPS-inducible	NN	O	B-protein
nuclear	NN	O	I-protein
factor-kappa	NN	O	I-protein
B/Rel	NN	O	I-protein
complex	NN	O	I-protein
)	NN	O	O
,	NN	O	O
that	NN	O	O
is	NN	O	O
(	NN	O	O
are	NN	O	O
)	NN	O	O
required	NN	O	O
for	NN	O	O
maximal	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
importance	NN	O	O
of	NN	O	O
resolution	NN	O	O
of	NN	O	O
this	NN	O	O
controversy	NN	O	O
is	NN	O	O
that	NN	O	O
future	NN	O	O
research	NN	O	O
in	NN	O	O
this	NN	O	O
area	NN	O	O
should	NN	O	O
be	NN	O	O
directed	NN	O	O
toward	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
alternative	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
can	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
successful	NN	O	O
production	NN	O	O
of	NN	O	O
NO	NN	O	O
.	NN	O	O

-DOCSTART-	O

X	NN	O	O
inactivation	NN	O	O
analysis	NN	O	O
in	NN	O	O
a	NN	O	O
female	NN	O	O
with	NN	O	O
hypomelanosis	NN	O	O
of	NN	O	O
Ito	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
balanced	NN	O	B-DNA
X	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
translocation	NN	O	I-DNA
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
functional	NN	O	O
disomy	NN	O	O
of	NN	O	O
Xp	NN	O	O
.	NN	O	O

X	NN	O	O
inactivation	NN	O	O
analysis	NN	O	O
was	NN	O	O
performed	NN	O	O
on	NN	O	O
normal	NN	O	O
and	NN	O	O
hypopigmented	NN	O	O
skin	NN	O	O
samples	NN	O	O
obtained	NN	O	O
from	NN	O	O
a	NN	O	O
female	NN	O	O
with	NN	O	O
hypomelanosis	NN	O	O
of	NN	O	O
Ito	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
balanced	NN	O	B-DNA
whole	NN	O	I-DNA
arm	NN	O	I-DNA
X	NN	O	I-DNA
;	NN	O	I-DNA
17	NN	O	I-DNA
translocation	NN	O	I-DNA
.	NN	O	O

Severe	NN	O	O
skewing	NN	O	O
of	NN	O	O
X	NN	O	O
inactivation	NN	O	O
resulting	NN	O	O
in	NN	O	O
inactivity	NN	O	O
of	NN	O	O
the	NN	O	O
intact	NN	O	O
X	NN	O	B-DNA
was	NN	O	O
found	NN	O	O
in	NN	O	O
blood	NN	O	O
and	NN	O	O
cultures	NN	O	O
of	NN	O	O
both	NN	O	O
types	NN	O	O
of	NN	O	O
skin	NN	O	O
,	NN	O	O
but	NN	O	O
analysis	NN	O	O
of	NN	O	O
DNA	NN	O	O
prepared	NN	O	O
directly	NN	O	O
from	NN	O	O
hypopigmented	NN	O	O
skin	NN	O	O
showed	NN	O	O
significant	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
translocated	NN	O	O
X	NN	O	B-DNA
,	NN	O	O
inconsistent	NN	O	O
with	NN	O	O
the	NN	O	O
usual	NN	O	O
mechanism	NN	O	O
of	NN	O	O
phenotypic	NN	O	O
expression	NN	O	O
in	NN	O	O
X	NN	O	B-DNA
;	NN	O	I-DNA
autosome	NN	O	I-DNA
translocations	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
dual	NN	O	O
colour	NN	O	O
FISH	NN	O	O
analysis	NN	O	O
using	NN	O	O
centromere	NN	O	O
specific	NN	O	O
probes	NN	O	O
for	NN	O	O
chromosomes	NN	O	O
X	NN	O	B-DNA
and	NN	O	O
17	NN	O	B-DNA
showed	NN	O	O
that	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
on	NN	O	O
both	NN	O	O
chromosomes	NN	O	B-DNA
lie	NN	O	O
within	NN	O	O
the	NN	O	O
alphoid	NN	O	O
arrays	NN	O	O
,	NN	O	O
making	NN	O	O
interruption	NN	O	O
of	NN	O	O
a	NN	O	O
locus	NN	O	O
on	NN	O	O
either	NN	O	O
chromosome	NN	O	O
unlikely	NN	O	O
.	NN	O	O

While	NN	O	O
partial	NN	O	O
variable	NN	O	O
monosomy	NN	O	O
of	NN	O	O
loci	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
17p	NN	O	I-DNA
can	NN	O	O
not	NN	O	O
be	NN	O	O
excluded	NN	O	O
as	NN	O	O
contributing	NN	O	O
to	NN	O	O
the	NN	O	O
phenotype	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
argued	NN	O	O
that	NN	O	O
the	NN	O	O
major	NN	O	O
likely	NN	O	O
factor	NN	O	O
is	NN	O	O
partial	NN	O	O
functional	NN	O	O
disomy	NN	O	O
of	NN	O	O
sequences	NN	O	O
on	NN	O	O
Xp	NN	O	B-DNA
in	NN	O	O
cell	NN	O	O
lineages	NN	O	O
that	NN	O	O
have	NN	O	O
failed	NN	O	O
to	NN	O	O
inactivate	NN	O	O
the	NN	O	O
intact	NN	O	O
X	NN	O	B-DNA
chromosome	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
herpesvirus	NN	O	B-DNA
Saimiri	NN	O	I-DNA
cis-acting	NN	O	I-DNA
DNA	NN	O	I-DNA
fragment	NN	O	I-DNA
that	NN	O	O
permits	NN	O	O
stable	NN	O	O
replication	NN	O	O
of	NN	O	O
episomes	NN	O	O
in	NN	O	O
transformed	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Herpesvirus	NN	O	O
saimiri	NN	O	O
is	NN	O	O
a	NN	O	O
lymphotropic	NN	O	O
herpesvirus	NN	O	O
capable	NN	O	O
of	NN	O	O
immortalizing	NN	O	O
and	NN	O	O
transforming	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Immortalized	NN	O	O
and	NN	O	O
transformed	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
harbor	NN	O	O
several	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
genome	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
persisting	NN	O	B-DNA
genome	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
mapping	NN	O	O
of	NN	O	O
the	NN	O	O
cis-acting	NN	O	B-DNA
genetic	NN	O	I-DNA
cis-acting	NN	O	I-DNA
segment	NN	O	I-DNA
(	NN	O	O
oriP	NN	O	B-DNA
)	NN	O	O
required	NN	O	O
for	NN	O	O
viral	NN	O	O
episomal	NN	O	O
maintenance	NN	O	O
is	NN	O	O
reported	NN	O	O
here	NN	O	O
.	NN	O	O

Viral	NN	O	B-DNA
DNA	NN	O	I-DNA
fragments	NN	O	I-DNA
that	NN	O	O
potentially	NN	O	O
contain	NN	O	O
oriP	NN	O	B-DNA
were	NN	O	O
cloned	NN	O	O
into	NN	O	O
a	NN	O	O
plasmid	NN	O	B-DNA
that	NN	O	O
contains	NN	O	O
the	NN	O	O
hygromycin	NN	O	B-DNA
resistance	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

After	NN	O	O
several	NN	O	O
round	NN	O	O
of	NN	O	O
subcloning	NN	O	O
followed	NN	O	O
by	NN	O	O
transfection	NN	O	O
,	NN	O	O
oriP	NN	O	B-DNA
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
a	NN	O	O
1.955-kb	NN	O	B-DNA
viral	NN	O	I-DNA
segment	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
viral	NN	O	O
fragment	NN	O	O
permits	NN	O	O
stable	NN	O	O
plasmid	NN	O	B-DNA
replication	NN	O	O
without	NN	O	O
deletion	NN	O	O
or	NN	O	O
rearrangement	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
episomal	NN	O	O
maintenance	NN	O	O
without	NN	O	O
integration	NN	O	O
or	NN	O	O
recombination	NN	O	O
.	NN	O	O

The	NN	O	O
function	NN	O	O
of	NN	O	O
oriP	NN	O	B-DNA
depends	NN	O	O
on	NN	O	O
a	NN	O	O
trans-acting	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
genome	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
1.955-kb	NN	O	B-DNA
viral	NN	O	I-DNA
segment	NN	O	I-DNA
includes	NN	O	O
a	NN	O	O
dyad	NN	O	O
symmetry	NN	O	O
region	NN	O	O
located	NN	O	O
between	NN	O	O
two	NN	O	O
small	NN	O	B-DNA
nuclear	NN	O	I-DNA
RNA	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
dihydrofolate	NN	O	B-DNA
reductase	NN	O	I-DNA
gene	NN	O	I-DNA
homolog	NN	O	I-DNA
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
this	NN	O	O
oriP	NN	O	B-DNA
contains	NN	O	O
novel	NN	O	O
elements	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
other	NN	O	O
DNA	NN	O	O
viruses	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
anergized	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
express	NN	O	O
perturbed	NN	O	O
AP-1	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Anergy	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
mechanism	NN	O	O
to	NN	O	O
ensure	NN	O	O
antigen-specific	NN	O	O
tolerance	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
adult	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
,	NN	O	O
anergy	NN	O	O
has	NN	O	O
mainly	NN	O	O
been	NN	O	O
studied	NN	O	O
at	NN	O	O
the	NN	O	O
cellular	NN	O	O
level	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
the	NN	O	O
T-cell-activating	NN	O	B-protein
superantigen	NN	O	I-protein
staphylococcal	NN	O	B-protein
enterotoxin	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	O
SEA	NN	O	B-protein
)	NN	O	O
to	NN	O	O
investigate	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
T-lymphocyte	NN	O	O
anergy	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Injection	NN	O	O
of	NN	O	O
SEA	NN	O	B-protein
to	NN	O	O
adult	NN	O	O
mice	NN	O	O
activates	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
expressing	NN	O	O
certain	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
TCR	NN	O	I-protein
)	NN	O	I-protein
variable	NN	O	I-protein
region	NN	O	I-protein
beta-chain	NN	O	I-protein
families	NN	O	I-protein
and	NN	O	O
induces	NN	O	O
strong	NN	O	O
and	NN	O	O
rapid	NN	O	O
production	NN	O	O
of	NN	O	O
interleukin	NN	O	O
2	NN	O	O
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
repeated	NN	O	O
injections	NN	O	O
of	NN	O	O
SEA	NN	O	B-protein
cause	NN	O	O
CD4+	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
deletion	NN	O	O
and	NN	O	O
anergy	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
CD4+	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
characterized	NN	O	O
by	NN	O	O
reduced	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
at	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
levels	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
expression	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
and	NN	O	O
octamer	NN	O	B-protein
binding	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
which	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Large	NN	O	O
amounts	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
significant	NN	O	O
quantities	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
were	NN	O	O
induced	NN	O	O
in	NN	O	O
SEA-activated	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
spleen	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
Oct-1	NN	O	O
and	NN	O	O
Oct-2	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
similar	NN	O	O
in	NN	O	O
both	NN	O	O
resting	NN	O	O
and	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
anergic	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
contained	NN	O	O
severely	NN	O	O
reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
Fos/Jun-containing	NN	O	B-protein
NF-AT	NN	O	I-protein
complexes	NN	O	I-protein
but	NN	O	O
expressed	NN	O	O
significant	NN	O	O
amounts	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
Oct	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
after	NN	O	O
SEA	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Resolution	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
demonstrated	NN	O	O
predominant	NN	O	O
expression	NN	O	O
of	NN	O	O
p50-p65	NN	O	B-protein
heterodimers	NN	O	I-protein
in	NN	O	O
activated	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
anergic	NN	O	O
cells	NN	O	O
mainly	NN	O	O
expressed	NN	O	O
the	NN	O	O
transcriptionally	NN	O	O
inactive	NN	O	O
p50	NN	O	B-protein
homodimer	NN	O	I-protein
.	NN	O	O

These	NN	O	O
alterations	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
repression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
anergic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Fas	NN	O	B-protein
ligation	NN	O	O
induces	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
Jun	NN	O	B-protein
kinase	NN	O	I-protein
activation	NN	O	O
independently	NN	O	O
of	NN	O	O
CD45	NN	O	B-protein
and	NN	O	O
Lck	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Stimulation	NN	O	O
through	NN	O	O
the	NN	O	O
Fas/APO-1	NN	O	B-protein
receptor	NN	O	I-protein
results	NN	O	O
in	NN	O	O
apoptosis	NN	O	O
through	NN	O	O
an	NN	O	O
incompletely	NN	O	O
characterized	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

More	NN	O	O
is	NN	O	O
known	NN	O	O
regarding	NN	O	O
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
that	NN	O	O
occur	NN	O	O
after	NN	O	O
ligation	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
that	NN	O	O
TCR	NN	O	B-protein
stimulation	NN	O	O
requires	NN	O	O
both	NN	O	O
the	NN	O	O
membrane	NN	O	B-protein
tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
,	NN	O	O
CD45	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
Src-family	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
Lck	NN	O	B-protein
,	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
cellular	NN	O	O
activation	NN	O	O
.	NN	O	O

Although	NN	O	O
prior	NN	O	O
studies	NN	O	O
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
and	NN	O	O
phosphatases	NN	O	O
in	NN	O	O
Fas	NN	O	B-protein
signaling	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
Fas	NN	O	O
ligation	NN	O	O
induces	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
deficient	NN	O	O
in	NN	O	O
either	NN	O	O
CD45	NN	O	B-protein
or	NN	O	O
Lck	NN	O	B-protein
.	NN	O	O

Further	NN	O	O
,	NN	O	O
in	NN	O	O
normal	NN	O	O
and	NN	O	O
CD45-	NN	O	B-cell_line
or	NN	O	I-cell_line
Lck-deficient	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
Fas	NN	O	O
stimulation	NN	O	O
results	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
Jun	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
proposed	NN	O	O
mediator	NN	O	O
of	NN	O	O
stress	NN	O	O
activation	NN	O	O
pathways	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
endogenous	NN	O	O
ceramide	NN	O	O
release	NN	O	O
in	NN	O	O
Fas-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
stimulation	NN	O	O
with	NN	O	O
a	NN	O	O
synthetic	NN	O	O
ceramide	NN	O	O
analog	NN	O	O
results	NN	O	O
in	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
suggesting	NN	O	O
ceramide	NN	O	O
release	NN	O	O
occurs	NN	O	O
proximal	NN	O	O
to	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
Fas	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
although	NN	O	O
CD45	NN	O	B-protein
and	NN	O	O
Lck	NN	O	B-protein
are	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
Fas	NN	O	B-protein
signaling	NN	O	O
,	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
transducing	NN	O	O
distal	NN	O	O
signals	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
after	NN	O	O
Fas	NN	O	B-protein
ligation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Globin	NN	O	O
gene	NN	O	O
switching	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
protein-DNA	NN	O	O
interactions	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
locus	NN	O	I-DNA
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
expressing	NN	O	O
the	NN	O	O
fetal	NN	O	O
or	NN	O	O
the	NN	O	O
adult	NN	O	O
globin	NN	O	B-DNA
gene	NN	O	I-DNA
program	NN	O	O
.	NN	O	O

To	NN	O	O
characterize	NN	O	O
the	NN	O	O
protein-DNA	NN	O	O
interactions	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
developmental	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
locus	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
by	NN	O	O
in	NN	O	O
vivo	NN	O	O
dimethyl	NN	O	O
sulfate	NN	O	O
footprinting	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
expressing	NN	O	O
either	NN	O	O
the	NN	O	O
fetal	NN	O	O
or	NN	O	O
the	NN	O	O
adult	NN	O	O
globin	NN	O	O
developmental	NN	O	O
program	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
locus	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
LCR	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
locus	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprints	NN	O	O
on	NN	O	O
NF-E2	NN	O	B-protein
(	NN	O	O
or	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
GATA-1	NN	O	B-DNA
motifs	NN	O	I-DNA
remained	NN	O	O
the	NN	O	O
same	NN	O	O
regardless	NN	O	O
of	NN	O	O
whether	NN	O	O
the	NN	O	O
fetal	NN	O	O
or	NN	O	O
the	NN	O	O
adult	NN	O	B-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
are	NN	O	O
expressed	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprints	NN	O	O
on	NN	O	O
GT	NN	O	B-DNA
(	NN	O	I-DNA
CACCC	NN	O	I-DNA
)	NN	O	I-DNA
motifs	NN	O	I-DNA
differed	NN	O	O
between	NN	O	O
the	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
fetal	NN	O	O
or	NN	O	O
the	NN	O	O
adult	NN	O	O
globin	NN	O	O
program	NN	O	O
.	NN	O	O

In	NN	O	O
promoter	NN	O	B-DNA
regions	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
actively	NN	O	B-DNA
transcribed	NN	O	I-DNA
genes	NN	O	I-DNA
demonstrated	NN	O	O
extensive	NN	O	O
and	NN	O	O
consistent	NN	O	O
footprints	NN	O	O
over	NN	O	O
the	NN	O	O
canonical	NN	O	O
elements	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
CACCC	NN	O	B-DNA
and	NN	O	I-DNA
CCAAT	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
adult	NN	O	B-cell_type
globin	NN	O	I-cell_type
expressing	NN	O	I-cell_type
cells	NN	O	I-cell_type
displayed	NN	O	O
more	NN	O	O
extensive	NN	O	O
footprints	NN	O	O
than	NN	O	O
the	NN	O	O
fetal	NN	O	B-cell_type
globin	NN	O	I-cell_type
expressing	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
regulatory	NN	O	I-DNA
sequences	NN	O	I-DNA
of	NN	O	O
both	NN	O	O
the	NN	O	O
Agamma-	NN	O	B-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
beta-globin	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
beta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
bulk	NN	O	O
of	NN	O	O
protein-DNA	NN	O	O
interactions	NN	O	O
that	NN	O	O
underlies	NN	O	O
the	NN	O	O
developmental	NN	O	O
control	NN	O	O
of	NN	O	O
globin	NN	O	B-DNA
genes	NN	O	I-DNA
takes	NN	O	O
place	NN	O	O
in	NN	O	O
the	NN	O	O
gamma-	NN	O	B-DNA
and	NN	O	I-DNA
beta-globin	NN	O	I-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
that	NN	O	O
GT	NN	O	B-DNA
motifs	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
locus	NN	O	I-DNA
LCR	NN	O	B-DNA
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
developmental	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	O
beta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
perhaps	NN	O	O
by	NN	O	O
increasing	NN	O	O
the	NN	O	O
probability	NN	O	O
of	NN	O	O
interaction	NN	O	O
of	NN	O	O
the	NN	O	O
LCR	NN	O	B-protein
holocomplex	NN	O	I-protein
with	NN	O	O
the	NN	O	O
fetal	NN	O	B-DNA
or	NN	O	I-DNA
the	NN	O	I-DNA
adult	NN	O	I-DNA
globin	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Antisense	NN	O	O
inhibition	NN	O	O
of	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
expression	NN	O	O
induces	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
monoblastoid	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
active	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
metabolite	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxycholecalciferol	NN	O	O
(	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
)	NN	O	O
acts	NN	O	O
as	NN	O	O
an	NN	O	O
antiproliferative	NN	O	O
and	NN	O	O
differentiating	NN	O	O
agent	NN	O	O
for	NN	O	O
the	NN	O	O
monoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U937	NN	O	I-cell_line
and	NN	O	O
as	NN	O	O
an	NN	O	O
important	NN	O	O
immunologic	NN	O	O
mediator	NN	O	O
implicated	NN	O	O
particularly	NN	O	O
in	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
cells	NN	O	O
belonging	NN	O	O
to	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
effects	NN	O	O
are	NN	O	O
controlled	NN	O	O
by	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

The	NN	O	O
objective	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
develop	NN	O	O
U937	NN	O	B-cell_line
transfectants	NN	O	O
expressing	NN	O	O
antisense	NN	O	O
VDR	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
to	NN	O	O
use	NN	O	O
these	NN	O	O
to	NN	O	O
examine	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
-VDR	NN	O	B-protein
interaction	NN	O	O
in	NN	O	O
this	NN	O	O
lineage	NN	O	O
.	NN	O	O

A	NN	O	O
2-kb	NN	O	B-DNA
VDR	NN	O	I-DNA
cDNA	NN	O	I-DNA
insert	NN	O	I-DNA
(	NN	O	O
including	NN	O	O
the	NN	O	O
complete	NN	O	O
VDR	NN	O	B-DNA
coding	NN	O	I-DNA
region	NN	O	I-DNA
)	NN	O	O
was	NN	O	O
cloned	NN	O	O
in	NN	O	O
an	NN	O	O
antisense	NN	O	O
orientation	NN	O	O
into	NN	O	O
the	NN	O	O
EBV	NN	O	O
episomal	NN	O	O
vector	NN	O	O
pMEP4	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
transfected	NN	O	O
into	NN	O	O
U937	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
resultant	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
DH42	NN	O	B-cell_line
,	NN	O	O
was	NN	O	O
hygromycin	NN	O	O
resistant	NN	O	O
,	NN	O	O
contained	NN	O	O
VDR	NN	O	B-DNA
cDNA	NN	O	I-DNA
,	NN	O	O
expressed	NN	O	O
fewer	NN	O	O
VDRs	NN	O	B-protein
than	NN	O	O
controls	NN	O	O
,	NN	O	O
and	NN	O	O
showed	NN	O	O
a	NN	O	O
substantial	NN	O	O
decrease	NN	O	O
in	NN	O	O
antiproliferative	NN	O	O
response	NN	O	O
to	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
increased	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
cells	NN	O	O
expressing	NN	O	O
macrophage	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
Ags	NN	O	I-protein
,	NN	O	O
including	NN	O	O
CD14	NN	O	B-protein
and	NN	O	O
CD11b	NN	O	B-protein
.	NN	O	O

A	NN	O	O
subpopulation	NN	O	O
of	NN	O	O
smaller	NN	O	O
cells	NN	O	O
did	NN	O	O
not	NN	O	O
express	NN	O	O
the	NN	O	O
differentiation	NN	O	O
markers	NN	O	O
after	NN	O	O
cadmium	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Cell	NN	O	O
cycle	NN	O	O
analysis	NN	O	O
showed	NN	O	O
shifts	NN	O	O
in	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
cells	NN	O	O
from	NN	O	O
G1	NN	O	O
to	NN	O	O
S	NN	O	O
phase	NN	O	O
,	NN	O	O
which	NN	O	O
were	NN	O	O
more	NN	O	O
pronounced	NN	O	O
after	NN	O	O
cadmium	NN	O	O
treatment	NN	O	O
.	NN	O	O

A	NN	O	O
considerable	NN	O	O
proportion	NN	O	O
of	NN	O	O
cells	NN	O	O
were	NN	O	O
outside	NN	O	O
the	NN	O	O
cycle	NN	O	O
and	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
confirmed	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
functional	NN	O	O
outcome	NN	O	O
of	NN	O	O
the	NN	O	O
VDR	NN	O	B-protein
antisense	NN	O	O
transfection	NN	O	O
suggests	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
myelomonocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
VDR	NN	O	B-protein
expression	NN	O	O
may	NN	O	O
act	NN	O	O
as	NN	O	O
a	NN	O	O
protective	NN	O	O
mechanism	NN	O	O
against	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
T	NN	O	O
lymphotropic	NN	O	O
virus-I	NN	O	O
infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
induces	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
beta-galactoside-binding	NN	O	B-protein
lectin	NN	O	I-protein
,	NN	O	O
galectin-3	NN	O	B-protein
.	NN	O	O

Animal	NN	O	O
lectins	NN	O	B-protein
play	NN	O	O
important	NN	O	O
roles	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
biological	NN	O	O
processes	NN	O	O
via	NN	O	O
their	NN	O	O
recognition	NN	O	O
of	NN	O	O
glycoconjugates	NN	O	O
.	NN	O	O

Galectin-3	NN	O	O
is	NN	O	O
a	NN	O	O
beta-galactoside-binding	NN	O	B-protein
lectin	NN	O	I-protein
previously	NN	O	O
designated	NN	O	O
as	NN	O	O
epsilon	NN	O	B-protein
BP	NN	O	I-protein
(	NN	O	O
IgE-binding	NN	O	B-protein
protein	NN	O	I-protein
)	NN	O	O
,	NN	O	O
CBP35	NN	O	B-protein
,	NN	O	O
Mac-2	NN	O	B-protein
,	NN	O	O
L-29	NN	O	B-protein
,	NN	O	O
and	NN	O	O
L-34	NN	O	B-protein
,	NN	O	O
and	NN	O	O
its	NN	O	O
expression	NN	O	O
has	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
various	NN	O	O
physiological	NN	O	O
and	NN	O	O
pathological	NN	O	O
processes	NN	O	O
,	NN	O	O
including	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
tumor	NN	O	O
transformation	NN	O	O
,	NN	O	O
and	NN	O	O
metastasis	NN	O	O
.	NN	O	O

Galectin-3	NN	O	B-protein
is	NN	O	O
widely	NN	O	O
distributed	NN	O	O
in	NN	O	O
various	NN	O	O
tissues	NN	O	O
and	NN	O	O
cell	NN	O	O
types	NN	O	O
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
many	NN	O	O
leukocytes	NN	O	B-cell_type
,	NN	O	O
with	NN	O	O
the	NN	O	O
notable	NN	O	O
exception	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
that	NN	O	O
galectin-3	NN	O	B-protein
is	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphotropic	NN	O	I-cell_line
virus	NN	O	I-cell_line
(	NN	O	I-cell_line
HTLV	NN	O	I-cell_line
)	NN	O	I-cell_line
-I-infected	NN	O	I-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
including	NN	O	O
F6T	NN	O	B-cell_line
,	NN	O	O
HUT	NN	O	B-cell_line
102	NN	O	I-cell_line
,	NN	O	O
K3T	NN	O	B-cell_line
,	NN	O	O
MT-2	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
SLB-I	NN	O	B-cell_line
,	NN	O	O
but	NN	O	O
is	NN	O	O
not	NN	O	O
expressed	NN	O	O
in	NN	O	O
non-HTLV-I-infected	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
such	NN	O	O
as	NN	O	O
Jurkat	NN	O	B-cell_line
,	NN	O	O
CEM	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
MOLT-4	NN	O	B-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
galectin-3	NN	O	B-protein
level	NN	O	O
was	NN	O	O
markedly	NN	O	O
increased	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
after	NN	O	O
infection	NN	O	O
with	NN	O	O
HTLV-I	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
uninfected	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
galectin-3	NN	O	B-protein
expression	NN	O	O
appeared	NN	O	O
to	NN	O	O
correlate	NN	O	O
well	NN	O	O
with	NN	O	O
HTLV-I	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
as	NN	O	O
undetectable	NN	O	O
or	NN	O	O
very	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
galectin-3	NN	O	B-protein
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
S1T	NN	O	B-cell_line
and	NN	O	I-cell_line
ATL-1K	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
are	NN	O	O
nonproductively	NN	O	O
infected	NN	O	O
with	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

In	NN	O	O
co-transfection	NN	O	O
experiments	NN	O	O
,	NN	O	O
the	NN	O	O
galectin-3	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
significantly	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
expression	NN	O	B-DNA
vectors	NN	O	I-DNA
encoding	NN	O	O
the	NN	O	O
40-kd	NN	O	B-protein
Tax	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
a	NN	O	O
potent	NN	O	O
transactivator	NN	O	O
in	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
various	NN	O	O
Tax	NN	O	B-protein
mutants	NN	O	I-protein
suggested	NN	O	O
that	NN	O	O
galectin-3	NN	O	B-DNA
promoter	NN	O	I-DNA
induction	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
cyclic-AMP-responsive	NN	O	B-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein/activation	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B/Rel	NN	O	I-protein
induction	NN	O	O
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
U-937	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
an	NN	O	O
HTLV-I	NN	O	O
Tax	NN	O	O
expression	NN	O	O
vector	NN	O	O
induced	NN	O	O
galectin-3	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
this	NN	O	O
cell	NN	O	O
line	NN	O	O
.	NN	O	O

Functionally	NN	O	O
,	NN	O	O
galectin-3	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
activate	NN	O	O
interleukin-2	NN	O	B-protein
production	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
raise	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
HTLV-I	NN	O	O
Tax	NN	O	B-protein
production	NN	O	O
induces	NN	O	O
the	NN	O	O
transcription	NN	O	O
and	NN	O	O
subsequent	NN	O	O
synthesis	NN	O	O
and	NN	O	O
secretion	NN	O	O
of	NN	O	O
galectin-3	NN	O	B-protein
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
may	NN	O	O
further	NN	O	O
activate	NN	O	O
these	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
altered	NN	O	O
properties	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
found	NN	O	O
in	NN	O	O
adult	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
induced	NN	O	O
by	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

-DOCSTART-	O

Monocytic	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
collagenase	NN	O	B-protein
.	NN	O	O

Interstitial	NN	O	O
collagenase	NN	O	B-protein
(	NN	O	O
MMP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
metalloproteinase	NN	O	B-protein
produced	NN	O	O
by	NN	O	O
resident	NN	O	B-cell_type
and	NN	O	I-cell_type
inflammatory	NN	O	I-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
connective	NN	O	O
tissue	NN	O	O
turnover	NN	O	O
,	NN	O	O
cleaves	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
collagen	NN	O	I-protein
fibrils	NN	O	I-protein
.	NN	O	O

This	NN	O	O
catalytic	NN	O	O
event	NN	O	O
is	NN	O	O
rate	NN	O	O
limiting	NN	O	O
in	NN	O	O
remodeling	NN	O	O
of	NN	O	O
tissues	NN	O	O
rich	NN	O	O
in	NN	O	O
fibrillar	NN	O	B-protein
collagen	NN	O	I-protein
such	NN	O	O
as	NN	O	O
the	NN	O	O
skin	NN	O	O
and	NN	O	O
lungs	NN	O	O
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
collagenase	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
cell-type	NN	O	O
specific	NN	O	O
;	NN	O	O
bacterial	NN	O	O
LPS	NN	O	O
and	NN	O	O
zymosan	NN	O	O
,	NN	O	O
a	NN	O	O
yeast	NN	O	O
cell	NN	O	O
wall	NN	O	O
derivative	NN	O	O
,	NN	O	O
are	NN	O	O
potent	NN	O	O
inducers	NN	O	O
of	NN	O	O
collagenase	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
do	NN	O	O
not	NN	O	O
alter	NN	O	O
fibroblast	NN	O	B-cell_type
collagenase	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Since	NN	O	O
promoter	NN	O	O
elements	NN	O	O
controlling	NN	O	O
collagenase	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
monocytic	NN	O	O
cells	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
previously	NN	O	O
defined	NN	O	O
,	NN	O	O
we	NN	O	O
sought	NN	O	O
to	NN	O	O
delineate	NN	O	O
responsive	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
collagenase	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
human	NN	O	B-cell_line
(	NN	O	I-cell_line
U937	NN	O	I-cell_line
)	NN	O	I-cell_line
and	NN	O	I-cell_line
murine	NN	O	I-cell_line
(	NN	O	I-cell_line
J774	NN	O	I-cell_line
)	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Deletion	NN	O	O
constructs	NN	O	O
containing	NN	O	O
as	NN	O	O
little	NN	O	O
as	NN	O	O
72	NN	O	O
bp	NN	O	O
of	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
-flanking	NN	O	I-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
collagenase	NN	O	B-DNA
promoter	NN	O	I-DNA
were	NN	O	O
sufficient	NN	O	O
for	NN	O	O
LPS-	NN	O	O
or	NN	O	O
zymosan-mediated	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
,	NN	O	O
whereas	NN	O	O
phorbol	NN	O	O
inducibility	NN	O	O
exhibited	NN	O	O
an	NN	O	O
absolute	NN	O	O
requirement	NN	O	O
for	NN	O	O
upstream	NN	O	O
elements	NN	O	O
including	NN	O	O
the	NN	O	O
polyoma	NN	O	B-DNA
enhancer	NN	O	I-DNA
A-binding	NN	O	I-DNA
protein-3	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
-83	NN	O	B-DNA
to	NN	O	I-DNA
-91	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
TTCA	NN	O	B-DNA
sequence	NN	O	I-DNA
(	NN	O	O
-102	NN	O	B-DNA
to	NN	O	I-DNA
-105	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
both	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

Mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
activator	NN	O	B-DNA
protein-1	NN	O	I-DNA
[	NN	O	I-DNA
AP-1	NN	O	I-DNA
]	NN	O	I-DNA
site	NN	O	I-DNA
at	NN	O	O
-72	NN	O	B-DNA
abolished	NN	O	O
basal	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
and	NN	O	O
LPS/zymosan	NN	O	O
inducibility	NN	O	O
,	NN	O	O
while	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
an	NN	O	O
NF-kappaB-like	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
-20	NN	O	O
to	NN	O	O
-10	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
LPS	NN	O	O
-and	NN	O	O
zymosan-treated	NN	O	B-cell_type
cells	NN	O	I-cell_type
showed	NN	O	O
strong	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
gel-shift	NN	O	O
analysis	NN	O	O
,	NN	O	O
and	NN	O	O
supershift	NN	O	O
analysis	NN	O	O
showed	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
complexes	NN	O	I-protein
contained	NN	O	O
specific	NN	O	O
members	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
jun	NN	O	B-DNA
and	NN	O	I-DNA
fos	NN	O	I-DNA
gene	NN	O	I-DNA
families	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
most	NN	O	O
LPS	NN	O	O
effects	NN	O	O
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
,	NN	O	O
mediates	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
collagenase	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
macrophagelike	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
regulation	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
,	NN	O	O
collagenase	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
cell	NN	O	O
wall	NN	O	O
derivatives	NN	O	O
of	NN	O	O
bacteria	NN	O	O
or	NN	O	O
yeast	NN	O	O
is	NN	O	O
largely	NN	O	O
independent	NN	O	O
of	NN	O	O
upstream	NN	O	B-DNA
promoter	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
in	NN	O	O
bacterial	NN	O	B-cell_type
lipopolysaccharide-stimulated	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
by	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
induces	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
that	NN	O	O
encode	NN	O	O
for	NN	O	O
proinflammatory	NN	O	B-protein
regulators	NN	O	I-protein
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Previous	NN	O	O
work	NN	O	O
has	NN	O	O
suggested	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
one	NN	O	O
LPS-induced	NN	O	O
event	NN	O	O
that	NN	O	O
mediates	NN	O	O
this	NN	O	O
response	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
this	NN	O	O
notion	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
LPS	NN	O	O
stimulated	NN	O	O
AP-1	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
transfected	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
macrophage	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
RAW	NN	O	I-cell_line
264.7	NN	O	I-cell_line
.	NN	O	O

As	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
phosphorylates	NN	O	O
and	NN	O	O
subsequently	NN	O	O
increases	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
c-Jun	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
whether	NN	O	O
LPS	NN	O	O
treatment	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
resulted	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
kinase	NN	O	B-protein
.	NN	O	O

LPS	NN	O	O
treatment	NN	O	O
of	NN	O	O
RAW	NN	O	B-cell_line
264.7	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
murine	NN	O	B-cell_type
bone	NN	O	I-cell_type
marrow-derived	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
resulted	NN	O	O
in	NN	O	O
rapid	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
p46	NN	O	B-protein
and	NN	O	I-protein
p54	NN	O	I-protein
isoforms	NN	O	I-protein
of	NN	O	O
JNK	NN	O	B-protein
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
wild-type	NN	O	O
and	NN	O	O
rough	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
LPS	NN	O	O
and	NN	O	O
synthetic	NN	O	O
lipid	NN	O	O
A	NN	O	O
resulted	NN	O	O
in	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
while	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
inhibitor	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
inhibited	NN	O	O
this	NN	O	O
response	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
LPS-	NN	O	O
LPS	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
LBP	NN	O	I-protein
)	NN	O	I-protein
complexes	NN	O	I-protein
to	NN	O	O
CD14	NN	O	B-protein
,	NN	O	O
a	NN	O	O
surface	NN	O	B-protein
receptor	NN	O	I-protein
that	NN	O	O
mediates	NN	O	O
many	NN	O	O
LPS	NN	O	O
responses	NN	O	O
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
crucial	NN	O	O
,	NN	O	O
as	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
60b	NN	O	I-protein
,	NN	O	O
which	NN	O	O
blocks	NN	O	O
this	NN	O	O
binding	NN	O	O
,	NN	O	O
inhibited	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
LPS	NN	O	O
activation	NN	O	O
of	NN	O	O
JNK	NN	O	B-protein
in	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
a	NN	O	O
CD14-	NN	O	O
and	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation-dependent	NN	O	O
event	NN	O	O
that	NN	O	O
may	NN	O	O
mediate	NN	O	O
the	NN	O	O
early	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
regulating	NN	O	O
LPS-triggered	NN	O	O
gene	NN	O	O
induction	NN	O	O
.	NN	O	O

-DOCSTART-	O

Surfactant	NN	O	O
suppresses	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
biophysical	NN	O	O
properties	NN	O	O
,	NN	O	O
pulmonary	NN	O	O
surfactant	NN	O	O
has	NN	O	O
immunomodulatory	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
both	NN	O	O
synthetic	NN	O	O
(	NN	O	O
Exosurf	NN	O	O
)	NN	O	O
and	NN	O	O
modified	NN	O	O
natural	NN	O	O
surfactant	NN	O	O
(	NN	O	O
Survanta	NN	O	O
)	NN	O	O
downregulated	NN	O	O
endotoxin-stimulated	NN	O	O
inflammatory	NN	O	B-RNA
c	NN	O	I-RNA
ytokine	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
and	NN	O	O
protein	NN	O	B-protein
products	NN	O	I-protein
(	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
[	NN	O	O
TNF	NN	O	O
]	NN	O	O
,	NN	O	O
interleukin-1-beta	NN	O	B-protein
[	NN	O	O
IL-1	NN	O	B-protein
]	NN	O	O
,	NN	O	O
interleukin-6	NN	O	B-protein
[	NN	O	O
IL-6	NN	O	B-protein
]	NN	O	O
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
both	NN	O	O
Exosurf	NN	O	O
and	NN	O	O
Survanta	NN	O	O
suppress	NN	O	O
TNF	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
secretion	NN	O	O
(	NN	O	O
85	NN	O	O
+/-	NN	O	O
4	NN	O	O
%	NN	O	O
mean	NN	O	O
percent	NN	O	O
inhibition	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
by	NN	O	O
Exosurf	NN	O	O
;	NN	O	O
71	NN	O	O
+/-	NN	O	O
6	NN	O	O
%	NN	O	O
by	NN	O	O
Survanta	NN	O	O
)	NN	O	O
by	NN	O	O
endotoxin-stimulated	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Because	NN	O	O
surfactant	NN	O	O
downregulated	NN	O	O
inflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
production	NN	O	O
similarly	NN	O	O
in	NN	O	O
both	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
THP-1	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
used	NN	O	O
this	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
to	NN	O	O
investigate	NN	O	O
whether	NN	O	O
surfactant	NN	O	O
affected	NN	O	O
transcriptional	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

Specifically	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
activation	NN	O	O
because	NN	O	O
it	NN	O	O
is	NN	O	O
crucial	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
many	NN	O	O
inflammatory	NN	O	B-DNA
cytokine	NN	O	I-DNA
genes	NN	O	I-DNA
including	NN	O	O
TNF	NN	O	B-DNA
,	NN	O	O
IL-1	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
IL-6	NN	O	B-DNA
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
both	NN	O	O
surfactants	NN	O	O
decreased	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
both	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
p50	NN	O	B-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
components	NN	O	I-protein
in	NN	O	O
LPS-activated	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
specific	NN	O	O
antibody	NN	O	O
induction	NN	O	O
of	NN	O	O
supershifts	NN	O	O
in	NN	O	O
mobility	NN	O	O
assays	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
are	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
suggest	NN	O	O
that	NN	O	O
surfactant	NN	O	O
's	NN	O	O
suppressive	NN	O	O
effects	NN	O	O
on	NN	O	O
inflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
production	NN	O	O
may	NN	O	O
involve	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
through	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
cell	NN	O	B-DNA
type-specific	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
B7.1	NN	O	I-DNA
gene	NN	O	I-DNA
regulated	NN	O	O
by	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
costimulatory	NN	O	B-protein
molecule	NN	O	I-protein
B7.1	NN	O	I-protein
provides	NN	O	O
a	NN	O	O
second	NN	O	O
signal	NN	O	O
critical	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
distribution	NN	O	O
of	NN	O	O
this	NN	O	O
integral	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
certain	NN	O	O
tissues	NN	O	O
where	NN	O	O
its	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
is	NN	O	O
modulated	NN	O	O
by	NN	O	O
multiple	NN	O	O
exogenous	NN	O	O
stimuli	NN	O	O
.	NN	O	O

To	NN	O	O
identify	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
specificity	NN	O	O
and	NN	O	O
inducibility	NN	O	O
,	NN	O	O
the	NN	O	O
chromatin	NN	O	B-DNA
configuration	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
B7.1	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
examined	NN	O	O
in	NN	O	O
intact	NN	O	O
nuclei	NN	O	O
from	NN	O	O
various	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
tissue-specific	NN	O	O
deoxyribonuclease	NN	O	B-DNA
I	NN	O	I-DNA
hypersensitive	NN	O	I-DNA
site	NN	O	I-DNA
approximately	NN	O	O
3kb	NN	O	B-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
led	NN	O	O
to	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
a	NN	O	O
cell	NN	O	B-DNA
type-specific	NN	O	I-DNA
enhancer	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
183-bp	NN	O	B-DNA
region	NN	O	I-DNA
was	NN	O	O
both	NN	O	O
cell	NN	O	O
type	NN	O	O
specific	NN	O	O
and	NN	O	O
responsive	NN	O	O
to	NN	O	O
two	NN	O	O
distinct	NN	O	O
stimuli	NN	O	O
,	NN	O	O
lipopolysaccharide	NN	O	O
and	NN	O	O
dibutyryl	NN	O	O
cAMP	NN	O	O
,	NN	O	O
known	NN	O	O
to	NN	O	O
regulate	NN	O	O
B7.1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Deletional	NN	O	O
and	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
revealed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
multiple	NN	O	O
functionally	NN	O	O
critical	NN	O	O
cis	NN	O	B-DNA
elements	NN	O	I-DNA
within	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
was	NN	O	O
a	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
NF	NN	O	I-DNA
)	NN	O	I-DNA
-kappaB	NN	O	I-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
B7.1-positive	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
this	NN	O	O
element	NN	O	O
bound	NN	O	O
several	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
family	NN	O	I-protein
,	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
already	NN	O	O
implicated	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
relevant	NN	O	O
to	NN	O	O
B7.1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
description	NN	O	O
,	NN	O	O
to	NN	O	O
our	NN	O	O
knowledge	NN	O	O
,	NN	O	O
of	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
that	NN	O	O
control	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
gene	NN	O	O
encoding	NN	O	O
a	NN	O	O
B7	NN	O	B-protein
costimulatory	NN	O	I-protein
molecule	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Ubiquitinylation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
c-Jun	NN	O	B-protein
and	NN	O	O
c-Fos	NN	O	B-protein
using	NN	O	O
reconstituted	NN	O	B-protein
ubiquitinylating	NN	O	I-protein
enzymes	NN	O	I-protein
.	NN	O	O

Recombinant	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
c-Fos	NN	O	B-protein
were	NN	O	O
ubiquitinylated	NN	O	O
by	NN	O	O
the	NN	O	O
ubiquitin	NN	O	B-protein
carrier	NN	O	I-protein
enzymes	NN	O	I-protein
E214K	NN	O	B-protein
,	NN	O	O
E220K	NN	O	B-protein
,	NN	O	O
or	NN	O	O
E232K	NN	O	B-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
ubiquitin-activating	NN	O	B-protein
enzyme	NN	O	I-protein
,	NN	O	O
E1	NN	O	B-protein
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
ubiquitin	NN	O	B-protein
protein	NN	O	I-protein
ligase	NN	O	I-protein
E3	NN	O	I-protein
substantially	NN	O	O
enhanced	NN	O	O
the	NN	O	O
E214K	NN	O	B-protein
-mediated	NN	O	O
ubiquitinylation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
c-Fos	NN	O	B-protein
.	NN	O	O

Truncated	NN	O	B-protein
c-Jun	NN	O	I-protein
and	NN	O	I-protein
c-Fos	NN	O	I-protein
mutant	NN	O	I-protein
proteins	NN	O	I-protein
including	NN	O	O
wbJun	NN	O	B-protein
and	NN	O	O
wbFos	NN	O	B-protein
were	NN	O	O
also	NN	O	O
ubiquitinylated	NN	O	O
under	NN	O	O
the	NN	O	O
same	NN	O	O
conditions	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
sites	NN	O	O
of	NN	O	O
ubiquitinylation	NN	O	O
are	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
dimerization	NN	O	B-protein
and	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
c-Fos	NN	O	B-protein
.	NN	O	O

The	NN	O	O
E3-dependent	NN	O	O
ubiquitinylation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
was	NN	O	O
inhibited	NN	O	O
upon	NN	O	O
the	NN	O	O
heterodimerization	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
with	NN	O	O
c-Fos	NN	O	B-protein
.	NN	O	O

Further	NN	O	O
addition	NN	O	O
of	NN	O	O
E220K	NN	O	B-protein
significantly	NN	O	O
enhanced	NN	O	O
ubiquitinylation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
in	NN	O	O
the	NN	O	O
heterodimer	NN	O	O
suggesting	NN	O	O
a	NN	O	O
regulatory	NN	O	O
role	NN	O	O
of	NN	O	O
E220K	NN	O	B-protein
.	NN	O	O

Polyubiquitinylated	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
wbFos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
wbJun	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
E220K-ubiquitinylated	NN	O	B-protein
c-Jun	NN	O	I-protein
,	NN	O	O
were	NN	O	O
readily	NN	O	O
degraded	NN	O	O
by	NN	O	O
the	NN	O	O
ATP-dependent	NN	O	B-protein
26	NN	O	I-protein
S	NN	O	I-protein
multicatalytic	NN	O	I-protein
proteases	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
temporal	NN	O	O
control	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
c-Fos	NN	O	B-protein
may	NN	O	O
be	NN	O	O
regulated	NN	O	O
through	NN	O	O
the	NN	O	O
ubiquitinylation	NN	O	O
pathways	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
ubiquitinylation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
c-Fos	NN	O	B-protein
may	NN	O	O
in	NN	O	O
turn	NN	O	O
be	NN	O	O
regulated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
heterodimerization	NN	O	O
between	NN	O	O
them	NN	O	O
and	NN	O	O
the	NN	O	O
cooperation	NN	O	O
between	NN	O	O
E220K	NN	O	B-protein
and	NN	O	O
E3	NN	O	O
mediated	NN	O	O
polyubiquitinylation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
B-cell	NN	O	I-protein
specific	NN	O	I-protein
transcriptional	NN	O	I-protein
coactivator	NN	O	I-protein
Bob1	NN	O	B-protein
.	NN	O	O

From	NN	O	O
a	NN	O	O
murine	NN	O	B-DNA
B-cell	NN	O	I-DNA
cDNA-library	NN	O	I-DNA
we	NN	O	O
have	NN	O	O
cloned	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
encoding	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
B-cell	NN	O	I-protein
specific	NN	O	I-protein
coactivator	NN	O	I-protein
mBob1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
protein	NN	O	O
is	NN	O	O
the	NN	O	O
murine	NN	O	O
homologue	NN	O	O
to	NN	O	O
the	NN	O	O
recently	NN	O	O
described	NN	O	O
human	NN	O	B-protein
coactivator	NN	O	I-protein
Bob1	NN	O	I-protein
(	NN	O	O
hBob1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
also	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
OBF-1	NN	O	B-protein
or	NN	O	O
OCA-B	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
also	NN	O	O
characterized	NN	O	O
the	NN	O	O
genomic	NN	O	B-protein
mBob1	NN	O	I-protein
clone	NN	O	I-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
its	NN	O	O
intron-exon	NN	O	O
structure	NN	O	O
has	NN	O	O
allowed	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
C-terminal	NN	O	B-protein
splice	NN	O	I-protein
variant	NN	O	I-protein
.	NN	O	O

mBob1	NN	O	B-protein
is	NN	O	O
B-cell	NN	O	O
restricted	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
all	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
representing	NN	O	O
different	NN	O	O
stages	NN	O	O
of	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

mBob1	NN	O	B-protein
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
octamer	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
and	NN	O	O
stimulates	NN	O	O
transcription	NN	O	O
mediated	NN	O	O
by	NN	O	O
these	NN	O	O
factors	NN	O	O
.	NN	O	O

-DOCSTART-	O

HIV	NN	O	B-protein
glycoprotein	NN	O	I-protein
120	NN	O	I-protein
enhances	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
glial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Involvement	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
/signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
well	NN	O	O
established	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
major	NN	O	O
glial	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
(	NN	O	O
CNS	NN	O	O
)	NN	O	O
,	NN	O	O
astrocytes	NN	O	B-cell_type
and	NN	O	O
microglia	NN	O	B-cell_type
,	NN	O	O
are	NN	O	O
key	NN	O	O
participants	NN	O	O
in	NN	O	O
mediating	NN	O	O
the	NN	O	O
neurologic	NN	O	O
dysfunction	NN	O	O
associated	NN	O	O
with	NN	O	O
HIV	NN	O	O
infection	NN	O	O
of	NN	O	O
the	NN	O	O
CNS	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-protein
envelope	NN	O	I-protein
glycoprotein	NN	O	I-protein
of	NN	O	O
HIV	NN	O	O
,	NN	O	O
glycoprotein	NN	O	B-protein
120	NN	O	I-protein
(	NN	O	O
gp120	NN	O	B-protein
)	NN	O	O
,	NN	O	O
to	NN	O	O
regulate	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
in	NN	O	O
glial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
because	NN	O	O
ICAM-1	NN	O	B-protein
is	NN	O	O
important	NN	O	O
in	NN	O	O
mediating	NN	O	O
immune	NN	O	O
responsiveness	NN	O	O
in	NN	O	O
the	NN	O	O
CNS	NN	O	O
,	NN	O	O
facilitating	NN	O	O
entry	NN	O	O
of	NN	O	O
HIV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
the	NN	O	O
CNS	NN	O	O
,	NN	O	O
and	NN	O	O
promoting	NN	O	O
syncytia	NN	O	O
formation	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
gp120	NN	O	B-protein
enhances	NN	O	O
ICAM-1	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
rat	NN	O	I-cell_type
astrocytes	NN	O	I-cell_type
,	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
astrocytes	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
astroglioma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
CRT	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
primary	NN	O	B-cell_type
rat	NN	O	I-cell_type
microglia	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
involved	NN	O	O
in	NN	O	O
gp120	NN	O	B-protein
-mediated	NN	O	O
enhancement	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
appear	NN	O	O
to	NN	O	O
involve	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
and	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
because	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
and	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
abrogate	NN	O	O
gp120	NN	O	B-protein
-mediated	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
both	NN	O	O
astrocytes	NN	O	B-cell_type
and	NN	O	O
microglia	NN	O	B-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
gp120	NN	O	B-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	O
STAT-1	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
JAK2	NN	O	B-protein
)	NN	O	O
in	NN	O	O
glial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
gp120	NN	O	B-protein
-mediated	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
has	NN	O	O
functional	NN	O	O
significance	NN	O	O
,	NN	O	O
as	NN	O	O
it	NN	O	O
enhances	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
bind	NN	O	O
to	NN	O	O
gp120-stimulated	NN	O	B-cell_type
human	NN	O	I-cell_type
astrocytes	NN	O	I-cell_type
in	NN	O	O
an	NN	O	O
ICAM-1	NN	O	B-protein
/beta	NN	O	O
2	NN	O	O
integrin	NN	O	B-protein
-dependent	NN	O	O
fashion	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
new	NN	O	O
insights	NN	O	O
into	NN	O	O
how	NN	O	O
gp120	NN	O	B-protein
can	NN	O	O
influence	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
glial	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
AIDS	NN	O	O
dementia	NN	O	O
complex	NN	O	O
.	NN	O	O

-DOCSTART-	O

IL-10	NN	O	B-protein
cooperates	NN	O	O
with	NN	O	O
TNF-alpha	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
HIV-1	NN	O	O
from	NN	O	O
latently	NN	O	B-cell_type
and	NN	O	I-cell_type
acutely	NN	O	I-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

IL-10	NN	O	B-protein
is	NN	O	O
elevated	NN	O	O
in	NN	O	O
HIV-1-infected	NN	O	O
individuals	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
disease	NN	O	O
progression	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
on	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
from	NN	O	O
infected	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

Although	NN	O	O
IL-10	NN	O	B-protein
alone	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
IL-10	NN	O	B-protein
markedly	NN	O	O
enhanced	NN	O	O
virion	NN	O	O
production	NN	O	O
from	NN	O	O
a	NN	O	O
chronically	NN	O	O
infected	NN	O	O
promonocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
U1	NN	O	B-cell_line
)	NN	O	O
and	NN	O	O
in	NN	O	O
acutely	NN	O	O
infected	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Neutralizing	NN	O	O
mAbs	NN	O	B-protein
to	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
indicated	NN	O	O
that	NN	O	O
both	NN	O	O
cytokines	NN	O	B-protein
were	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
and	NN	O	O
were	NN	O	O
required	NN	O	O
to	NN	O	O
generate	NN	O	O
a	NN	O	O
synergistic	NN	O	O
increase	NN	O	O
in	NN	O	O
virus	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
cytokines	NN	O	B-protein
were	NN	O	O
distinguishable	NN	O	O
functionally	NN	O	O
since	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
TNF-alpha	NN	O	B-protein
attenuated	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
cooperativity	NN	O	O
,	NN	O	O
while	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
IL-10	NN	O	B-protein
potentiated	NN	O	O
their	NN	O	O
cooperativity	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
play	NN	O	O
different	NN	O	O
roles	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
virus	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
Ab	NN	O	O
blocking	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
secretion	NN	O	O
studies	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
either	NN	O	O
endogenous	NN	O	B-protein
TNF-alpha	NN	O	I-protein
or	NN	O	O
IL-10	NN	O	B-protein
was	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
cooperativity	NN	O	O
,	NN	O	O
nor	NN	O	O
was	NN	O	O
an	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
combination	NN	O	O
with	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
IL-10	NN	O	B-protein
stimulated	NN	O	O
activating	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
binding	NN	O	O
activities	NN	O	O
and	NN	O	O
cooperated	NN	O	O
to	NN	O	O
increase	NN	O	O
HIV-1	NN	O	O
steady-state	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
and	NN	O	O
enhance	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
-directed	NN	O	O
transcription	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
IL-10	NN	O	B-protein
effect	NN	O	O
occurs	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
down-regulating	NN	O	O
the	NN	O	O
cellular	NN	O	O
immune	NN	O	O
response	NN	O	O
to	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
may	NN	O	O
also	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
TNF-alpha	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Interaction	NN	O	O
of	NN	O	O
HTLV-I	NN	O	B-protein
Tax	NN	O	I-protein
with	NN	O	O
the	NN	O	O
human	NN	O	B-protein
proteasome	NN	O	I-protein
:	NN	O	O
implications	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
induction	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
etiologically	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ATL	NN	O	O
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
degenerative	NN	O	O
neurologic	NN	O	O
syndrome	NN	O	O
termed	NN	O	O
tropical	NN	O	O
spastic	NN	O	O
paraparesis	NN	O	O
(	NN	O	O
TSP	NN	O	O
)	NN	O	O
.	NN	O	O

HTLV-I	NN	O	O
encodes	NN	O	O
a	NN	O	O
potent	NN	O	O
transactivator	NN	O	B-protein
protein	NN	O	O
termed	NN	O	O
Tax	NN	O	B-protein
that	NN	O	O
appears	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
T-cell	NN	O	O
immortalization	NN	O	O
.	NN	O	O

Even	NN	O	O
though	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
Tax	NN	O	B-protein
induces	NN	O	O
transformation	NN	O	O
are	NN	O	O
still	NN	O	O
unknown	NN	O	O
,	NN	O	O
it	NN	O	O
seems	NN	O	O
likely	NN	O	O
that	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
to	NN	O	O
alter	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
plays	NN	O	O
an	NN	O	O
important	NN	O	O
part	NN	O	O
in	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

Tax	NN	O	B-protein
does	NN	O	O
not	NN	O	O
bind	NN	O	O
directly	NN	O	O
to	NN	O	O
DNA	NN	O	O
but	NN	O	O
rather	NN	O	O
deregulates	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

One	NN	O	O
family	NN	O	O
of	NN	O	O
host	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
whose	NN	O	O
activity	NN	O	O
is	NN	O	O
altered	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
includes	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
.	NN	O	O

These	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
post-transcriptionally	NN	O	O
regulated	NN	O	O
by	NN	O	O
their	NN	O	O
assembly	NN	O	O
with	NN	O	O
a	NN	O	O
second	NN	O	O
family	NN	O	O
of	NN	O	O
inhibitory	NN	O	B-protein
proteins	NN	O	I-protein
termed	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
that	NN	O	O
serve	NN	O	O
to	NN	O	O
sequester	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

Upon	NN	O	O
cellular	NN	O	O
activation	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
is	NN	O	O
phosphorylated	NN	O	O
,	NN	O	O
polyubiquitinated	NN	O	O
,	NN	O	O
and	NN	O	O
degraded	NN	O	O
in	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
.	NN	O	O

This	NN	O	O
proteolytic	NN	O	O
event	NN	O	O
liberates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
permitting	NN	O	O
its	NN	O	O
rapid	NN	O	O
translocation	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
where	NN	O	O
it	NN	O	O
binds	NN	O	O
to	NN	O	O
its	NN	O	O
cognate	NN	O	B-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
the	NN	O	O
p105	NN	O	B-protein
precursor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	B-protein
subunit	NN	O	I-protein
is	NN	O	O
also	NN	O	O
post-translationally	NN	O	O
processed	NN	O	O
in	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
Tax	NN	O	B-protein
activates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
remain	NN	O	O
unclear	NN	O	O
,	NN	O	O
and	NN	O	O
findings	NN	O	O
presented	NN	O	O
in	NN	O	O
the	NN	O	O
literature	NN	O	O
are	NN	O	O
often	NN	O	O
controversial	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
physical	NN	O	O
interaction	NN	O	O
between	NN	O	O
Tax	NN	O	B-protein
and	NN	O	O
the	NN	O	O
HsN3	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
proteasome	NN	O	I-protein
.	NN	O	O

This	NN	O	O
raises	NN	O	O
the	NN	O	O
intriguing	NN	O	O
possibility	NN	O	O
that	NN	O	O
physical	NN	O	O
association	NN	O	O
of	NN	O	O
the	NN	O	O
HsN3	NN	O	B-protein
proteasome	NN	O	I-protein
subunit	NN	O	I-protein
with	NN	O	O
HTLV-I	NN	O	B-protein
Tax	NN	O	I-protein
coupled	NN	O	O
with	NN	O	O
the	NN	O	O
independent	NN	O	O
interaction	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
with	NN	O	O
either	NN	O	O
p100	NN	O	O
or	NN	O	O
p65-	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
targets	NN	O	O
these	NN	O	O
cytoplasmic	NN	O	B-protein
NF-kappa	NN	O	I-protein
B/Rel	NN	O	I-protein
complexes	NN	O	I-protein
to	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
for	NN	O	O
processing	NN	O	O
.	NN	O	O

-DOCSTART-	O

Immunophenotype	NN	O	O
of	NN	O	O
intraductal	NN	O	O
carcinoma	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
--	NN	O	O
Mammography	NN	O	O
and	NN	O	O
breast-conserving	NN	O	O
therapy	NN	O	O
have	NN	O	O
focused	NN	O	O
attention	NN	O	O
on	NN	O	O
the	NN	O	O
classification	NN	O	O
of	NN	O	O
intraductal	NN	O	O
carcinoma	NN	O	O
(	NN	O	O
IDC	NN	O	O
)	NN	O	O
and	NN	O	O
emphasized	NN	O	O
the	NN	O	O
prognostic	NN	O	O
importance	NN	O	O
of	NN	O	O
comedo	NN	O	O
versus	NN	O	O
noncomedo	NN	O	O
variants	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
histochemical	NN	O	O
markers	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
immunophenotype	NN	O	O
of	NN	O	O
43	NN	O	O
IDCs	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
comedo	NN	O	O
versus	NN	O	O
noncomedo	NN	O	O
status	NN	O	O
and	NN	O	O
patterns	NN	O	O
of	NN	O	O
angiogenesis	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
--	NN	O	O
Reactions	NN	O	O
in	NN	O	O
comedo	NN	O	O
carcinomas	NN	O	O
were	NN	O	O
significantly	NN	O	O
negative	NN	O	O
for	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
positive	NN	O	O
for	NN	O	O
p53	NN	O	B-protein
and	NN	O	O
HER-2/neu	NN	O	B-protein
more	NN	O	O
often	NN	O	O
than	NN	O	O
the	NN	O	O
noncomedo	NN	O	O
variant	NN	O	O
.	NN	O	O

All	NN	O	O
seven	NN	O	O
IDCs	NN	O	O
associated	NN	O	O
with	NN	O	O
Paget	NN	O	O
's	NN	O	O
disease	NN	O	O
showed	NN	O	O
positive	NN	O	O
reactions	NN	O	O
for	NN	O	O
HER-2/neu	NN	O	B-protein
.	NN	O	O

Basement	NN	O	O
membrane	NN	O	O
immunoreactivity	NN	O	O
for	NN	O	O
type	NN	O	B-protein
IV	NN	O	I-protein
collagen	NN	O	I-protein
and	NN	O	O
laminin	NN	O	B-protein
was	NN	O	O
discontinuous	NN	O	O
in	NN	O	O
most	NN	O	O
examples	NN	O	O
of	NN	O	O
IDC	NN	O	O
regardless	NN	O	O
of	NN	O	O
type	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
trend	NN	O	O
toward	NN	O	O
more	NN	O	O
intense	NN	O	O
staining	NN	O	O
in	NN	O	O
comedo	NN	O	O
than	NN	O	O
in	NN	O	O
noncomedo	NN	O	O
carcinomas	NN	O	O
.	NN	O	O

Periductal	NN	O	O
angiogenesis	NN	O	O
was	NN	O	O
not	NN	O	O
significantly	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
type	NN	O	O
of	NN	O	O
IDC	NN	O	O
but	NN	O	O
was	NN	O	O
more	NN	O	O
pronounced	NN	O	O
with	NN	O	O
comedo	NN	O	O
carcinomas	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
--	NN	O	O
These	NN	O	O
observations	NN	O	O
indicate	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
immunophenotypic	NN	O	O
correlates	NN	O	O
to	NN	O	O
the	NN	O	O
current	NN	O	O
structural	NN	O	O
classification	NN	O	O
of	NN	O	O
IDC	NN	O	O
.	NN	O	O

The	NN	O	O
immunophenotype	NN	O	O
of	NN	O	O
IDC	NN	O	O
is	NN	O	O
helpful	NN	O	O
in	NN	O	O
subclassifying	NN	O	O
an	NN	O	O
IDC	NN	O	O
and	NN	O	O
could	NN	O	O
prove	NN	O	O
useful	NN	O	O
as	NN	O	O
a	NN	O	O
prognostic	NN	O	O
indicator	NN	O	O
for	NN	O	O
local	NN	O	O
control	NN	O	O
in	NN	O	O
patients	NN	O	O
treated	NN	O	O
by	NN	O	O
breast-conserving	NN	O	O
therapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
phosphatase	NN	O	I-protein
2A	NN	O	I-protein
induces	NN	O	O
serine/threonine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
subcellular	NN	O	O
redistribution	NN	O	O
,	NN	O	O
and	NN	O	O
functional	NN	O	O
inhibition	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
.	NN	O	O

Signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	O
STATs	NN	O	B-protein
)	NN	O	O
are	NN	O	O
rapidly	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
residues	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
cytokine	NN	O	B-protein
and	NN	O	O
growth	NN	O	B-protein
factor	NN	O	I-protein
stimulation	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

STATs	NN	O	B-protein
hereafter	NN	O	O
are	NN	O	O
translocated	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
where	NN	O	O
they	NN	O	O
act	NN	O	O
as	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Recent	NN	O	O
reports	NN	O	O
suggest	NN	O	O
that	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STATs	NN	O	B-protein
also	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
-mediated	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
serine/threonine	NN	O	B-protein
phosphatases	NN	O	I-protein
in	NN	O	O
STAT3	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
antigen-specific	NN	O	I-cell_line
CD4	NN	O	I-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
cutaneous	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
expressing	NN	O	O
a	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
STAT3	NN	O	B-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
phosphatases	NN	O	I-protein
(	NN	O	O
PPs	NN	O	B-protein
)	NN	O	O
PP1/PP2A	NN	O	B-protein
,	NN	O	O
calyculin	NN	O	B-protein
A	NN	O	I-protein
,	NN	O	O
induces	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
on	NN	O	O
serine	NN	O	O
and	NN	O	O
threonine	NN	O	O
residues	NN	O	O
,	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
inhibition	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
relocation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
from	NN	O	O
the	NN	O	O
nucleus	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

Similar	NN	O	O
results	NN	O	O
were	NN	O	O
obtained	NN	O	O
with	NN	O	O
other	NN	O	O
PP2A	NN	O	O
inhibitors	NN	O	O
(	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
,	NN	O	O
endothall	NN	O	O
thioanhydride	NN	O	O
)	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
PP1	NN	O	B-protein
(	NN	O	O
tautomycin	NN	O	O
)	NN	O	O
or	NN	O	O
PP2B	NN	O	B-protein
(	NN	O	O
cyclosporine	NN	O	O
A	NN	O	O
)	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
with	NN	O	O
the	NN	O	O
broad	NN	O	O
serine/threonine	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
staurosporine	NN	O	O
partly	NN	O	O
blocked	NN	O	O
the	NN	O	O
calyculin	NN	O	B-protein
A	NN	O	I-protein
-induced	NN	O	O
STAT3	NN	O	B-protein
phosphorylation	NN	O	O
,	NN	O	O
whereas	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
serine/threonine	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase-1	NN	O	I-protein
extracellular-regulated	NN	O	I-protein
kinase-kinase	NN	O	I-protein
,	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
p38	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
,	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
we	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
PP2A	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
phosphorylation	NN	O	O
and	NN	O	O
subcellular	NN	O	O
distribution	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
phosphorylation	NN	O	O
is	NN	O	O
balanced	NN	O	O
between	NN	O	O
a	NN	O	O
staurosporine-sensitive	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
and	NN	O	O
PP2A	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Polyamines	NN	O	O
in	NN	O	O
human	NN	O	O
breast	NN	O	O
cancer	NN	O	O
and	NN	O	O
its	NN	O	O
relations	NN	O	O
to	NN	O	O
classical	NN	O	O
prognostic	NN	O	O
features	NN	O	O
:	NN	O	O
clinical	NN	O	O
implications	NN	O	O
.	NN	O	O

Experimental	NN	O	O
evidence	NN	O	O
suggest	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
of	NN	O	O
polyamines	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
development	NN	O	O
.	NN	O	O

Polyamines	NN	O	O
have	NN	O	O
been	NN	O	O
determined	NN	O	O
in	NN	O	O
tissue	NN	O	O
and	NN	O	O
erythrocyte	NN	O	O
samples	NN	O	O
from	NN	O	O
100	NN	O	O
patients	NN	O	O
with	NN	O	O
primary	NN	O	O
invasive	NN	O	O
breast	NN	O	O
cancer	NN	O	O
and	NN	O	O
30	NN	O	O
patients	NN	O	O
with	NN	O	O
fibroadenomas	NN	O	O
.	NN	O	O

Statistical	NN	O	O
analysis	NN	O	O
was	NN	O	O
performed	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
prognostic	NN	O	O
value	NN	O	O
of	NN	O	O
the	NN	O	O
polyamine	NN	O	O
patterns	NN	O	O
of	NN	O	O
tumor	NN	O	O
tissues	NN	O	O
and	NN	O	O
erythrocytes	NN	O	O
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
clinical	NN	O	O
and	NN	O	O
histological	NN	O	O
prognostic	NN	O	O
factors	NN	O	O
.	NN	O	O

In	NN	O	O
malignant	NN	O	O
tissues	NN	O	O
,	NN	O	O
polyamine	NN	O	O
levels	NN	O	O
were	NN	O	O
significantly	NN	O	O
higher	NN	O	O
than	NN	O	O
in	NN	O	O
benign	NN	O	O
tissues	NN	O	O
.	NN	O	O

They	NN	O	O
correlated	NN	O	O
with	NN	O	O
markers	NN	O	O
of	NN	O	O
tumor	NN	O	O
aggressivity	NN	O	O
(	NN	O	O
axillary	NN	O	O
node	NN	O	O
involvement	NN	O	O
and	NN	O	O
especially	NN	O	O
with	NN	O	O
markers	NN	O	O
of	NN	O	O
high	NN	O	O
mitotic	NN	O	O
rate	NN	O	O
as	NN	O	O
Ki-67	NN	O	O
staining	NN	O	O
,	NN	O	O
histological	NN	O	O
grade	NN	O	O
)	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
estrogen	NN	O	O
and	NN	O	O
progesterone	NN	O	O
status	NN	O	O
,	NN	O	O
tumor	NN	O	O
size	NN	O	O
and	NN	O	O
polyamine	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Erythrocyte	NN	O	O
polyamines	NN	O	O
levels	NN	O	O
were	NN	O	O
identical	NN	O	O
between	NN	O	O
cancer	NN	O	O
patients	NN	O	O
and	NN	O	O
controls	NN	O	O
.	NN	O	O

The	NN	O	O
knowledge	NN	O	O
of	NN	O	O
the	NN	O	O
polyamine	NN	O	O
pattern	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
could	NN	O	O
become	NN	O	O
useful	NN	O	O
in	NN	O	O
clinical	NN	O	O
practice	NN	O	O
particularly	NN	O	O
if	NN	O	O
polyamine	NN	O	O
metabolism	NN	O	O
is	NN	O	O
targeted	NN	O	O
as	NN	O	O
a	NN	O	O
therapeutic	NN	O	O
approach	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
triggered	NN	O	O
by	NN	O	O
the	NN	O	O
FcepsilonRIIb	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
CD23	NN	O	B-protein
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
lead	NN	O	O
to	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
reaction	NN	O	O
of	NN	O	O
allergic	NN	O	O
asthma	NN	O	O
.	NN	O	O

Alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
are	NN	O	O
activated	NN	O	O
when	NN	O	O
IgE/allergen	NN	O	B-protein
immune	NN	O	I-protein
complexes	NN	O	I-protein
bind	NN	O	O
to	NN	O	O
the	NN	O	O
CD23	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
We	NN	O	O
sought	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
regulating	NN	O	O
this	NN	O	O
early	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
focused	NN	O	O
on	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
triggered	NN	O	O
by	NN	O	O
CD23	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
promonocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U937	NN	O	I-cell_line
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
CD23	NN	O	B-protein
was	NN	O	O
cross-linked	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
IgE	NN	O	B-protein
immune	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Surface	NN	O	O
expression	NN	O	O
of	NN	O	O
CD23	NN	O	B-protein
was	NN	O	O
determined	NN	O	O
by	NN	O	O
FACS	NN	O	O
analysis	NN	O	O
.	NN	O	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
gene	NN	O	O
transcription	NN	O	O
were	NN	O	O
studied	NN	O	O
by	NN	O	O
gel-shift	NN	O	O
assays	NN	O	O
and	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

IkappaBalpha	NN	O	B-protein
phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
Western	NN	O	O
blot	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappaB	NN	O	I-protein
is	NN	O	O
the	NN	O	O
main	NN	O	O
transcription	NN	O	O
factor	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
activation	NN	O	O
that	NN	O	O
follows	NN	O	O
CD23	NN	O	B-protein
cross-linking	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

CD23-induced	NN	O	B-protein
NF-kappaB	NN	O	I-protein
is	NN	O	O
a	NN	O	O
heterodimer	NN	O	O
composed	NN	O	O
of	NN	O	O
p65/p50	NN	O	B-protein
subunits	NN	O	I-protein
.	NN	O	O

NF-kappaB	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
is	NN	O	O
secondary	NN	O	O
to	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
subsequent	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
inhibitory	NN	O	I-protein
molecule	NN	O	I-protein
IkappaBalpha	NN	O	B-protein
.	NN	O	O

Tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
-dependent	NN	O	O
,	NN	O	O
and	NN	O	O
not	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
-dependent	NN	O	O
,	NN	O	O
pathways	NN	O	O
mediate	NN	O	O
CD23	NN	O	B-protein
-triggered	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
but	NN	O	O
do	NN	O	O
not	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
direct	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
.	NN	O	O

IkappaBalpha	NN	O	B-protein
degradation	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
correlate	NN	O	O
with	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
TNF-alpha	NN	O	B-protein
and	NN	O	O
IL-1beta	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
NF-kappaB	NN	O	B-protein
is	NN	O	O
the	NN	O	O
main	NN	O	O
transcription	NN	O	O
factor	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
of	NN	O	O
CD23	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
TNFalpha	NN	O	B-protein
-mediated	NN	O	O
NFkappaB	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
myricetin	NN	O	O
and	NN	O	O
other	NN	O	O
flavonoids	NN	O	O
through	NN	O	O
downregulating	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
IKK	NN	O	B-protein
in	NN	O	O
ECV304	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Flavonoids	NN	O	O
are	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
naturally-occurring	NN	O	O
phenolic	NN	O	O
compounds	NN	O	O
in	NN	O	O
the	NN	O	O
plant	NN	O	O
kingdom	NN	O	O
,	NN	O	O
and	NN	O	O
many	NN	O	O
flavonoids	NN	O	O
are	NN	O	O
found	NN	O	O
with	NN	O	O
vascular	NN	O	O
protective	NN	O	O
properties	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
how	NN	O	O
the	NN	O	O
protective	NN	O	O
response	NN	O	O
is	NN	O	O
exerted	NN	O	O
by	NN	O	O
flavonoids	NN	O	O
is	NN	O	O
not	NN	O	O
well	NN	O	O
characterized	NN	O	O
.	NN	O	O

In	NN	O	O
view	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NFkappaB	NN	O	B-protein
)	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
atherosclerosis	NN	O	O
,	NN	O	O
prevention	NN	O	O
of	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
represents	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
protecting	NN	O	O
vascular	NN	O	O
injury	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
flavonoids	NN	O	O
on	NN	O	O
NFkappaB	NN	O	B-protein
/inhibitor-kappaB	NN	O	O
(	NN	O	O
IkappaB	NN	O	B-protein
)	NN	O	O
system	NN	O	O
in	NN	O	O
ECV304	NN	O	B-cell_line
cells	NN	O	I-cell_line
activated	NN	O	O
with	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNFalpha	NN	O	B-protein
)	NN	O	O
were	NN	O	O
examined	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
action	NN	O	O
of	NN	O	O
six	NN	O	O
flavonoids	NN	O	O
on	NN	O	O
IkappaB	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
IKK	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
,	NN	O	O
an	NN	O	O
enzyme	NN	O	O
recently	NN	O	O
found	NN	O	O
to	NN	O	O
phosphorylate	NN	O	O
critical	NN	O	O
serine	NN	O	O
residues	NN	O	O
of	NN	O	O
IkappaB	NN	O	O
for	NN	O	O
degradation	NN	O	O
.	NN	O	O

Of	NN	O	O
six	NN	O	O
flavonoids	NN	O	O
tested	NN	O	O
,	NN	O	O
myricetin	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
strongly	NN	O	O
inhibit	NN	O	O
IKK	NN	O	B-protein
kinase	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
prevent	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
and	NN	O	O
IkappaBbeta	NN	O	B-protein
in	NN	O	O
activated	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
myricetin	NN	O	O
was	NN	O	O
also	NN	O	O
found	NN	O	O
to	NN	O	O
inhibit	NN	O	O
NFkappaB	NN	O	B-protein
activity	NN	O	O
correlated	NN	O	O
with	NN	O	O
suppression	NN	O	O
of	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
ECV304	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Therefore	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
flavonoids	NN	O	O
may	NN	O	O
be	NN	O	O
of	NN	O	O
therapeutic	NN	O	O
value	NN	O	O
for	NN	O	O
vascular	NN	O	O
disease	NN	O	O
through	NN	O	O
down	NN	O	O
regulation	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
/IkappaB	NN	O	B-protein
system	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Wiley-Liss	NN	O	O
,	NN	O	O
Inc	NN	O	O
.	NN	O	O

-DOCSTART-	O

Bcl-2	NN	O	B-protein
-mediated	NN	O	O
drug	NN	O	O
resistance	NN	O	O
:	NN	O	O
inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
blocking	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
lymphocytes	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
-induced	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	O
transcription	NN	O	O
.	NN	O	O

Bcl-2	NN	O	B-protein
inhibits	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
stimuli	NN	O	O
,	NN	O	O
including	NN	O	O
chemotherapy	NN	O	O
drugs	NN	O	O
and	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
generally	NN	O	O
accepted	NN	O	O
that	NN	O	O
Bcl-2	NN	O	B-protein
exerts	NN	O	O
its	NN	O	O
antiapoptotic	NN	O	O
effects	NN	O	O
mainly	NN	O	O
by	NN	O	O
dimerizing	NN	O	O
with	NN	O	O
proapoptotic	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Bcl-2	NN	O	B-protein
family	NN	O	I-protein
such	NN	O	O
as	NN	O	O
Bax	NN	O	B-protein
and	NN	O	O
Bad	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
antiapoptotic	NN	O	O
effects	NN	O	O
is	NN	O	O
unclear	NN	O	O
.	NN	O	O

Paclitaxel	NN	O	O
and	NN	O	O
other	NN	O	O
drugs	NN	O	O
that	NN	O	O
disturb	NN	O	O
microtubule	NN	O	B-protein
dynamics	NN	O	O
kill	NN	O	O
cells	NN	O	O
in	NN	O	O
a	NN	O	O
Fas	NN	O	B-protein
/Fas	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
FasL	NN	O	B-protein
)	NN	O	O
-dependent	NN	O	O
manner	NN	O	O
;	NN	O	O
antibody	NN	O	O
to	NN	O	O
FasL	NN	O	B-protein
inhibits	NN	O	O
paclitaxel-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
Bcl-2	NN	O	B-protein
overexpression	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
prevention	NN	O	O
of	NN	O	O
chemotherapy	NN	O	O
(	NN	O	O
paclitaxel	NN	O	O
)	NN	O	O
-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
and	NN	O	O
blocks	NN	O	O
paclitaxel-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
this	NN	O	O
effect	NN	O	O
is	NN	O	O
that	NN	O	O
Bcl-2	NN	O	B-protein
prevents	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
(	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
lymphocytes	NN	O	I-protein
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
activated	NN	O	O
by	NN	O	O
microtubule	NN	O	B-protein
damage	NN	O	O
)	NN	O	O
by	NN	O	O
binding	NN	O	O
and	NN	O	O
sequestering	NN	O	O
calcineurin	NN	O	O
,	NN	O	O
a	NN	O	O
calcium-dependent	NN	O	O
phosphatase	NN	O	O
that	NN	O	O
must	NN	O	O
dephosphorylate	NN	O	O
NFAT	NN	O	B-protein
to	NN	O	O
move	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Without	NN	O	O
NFAT	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
,	NN	O	O
the	NN	O	O
FasL	NN	O	B-protein
gene	NN	O	O
is	NN	O	O
not	NN	O	O
transcribed	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
it	NN	O	O
appears	NN	O	O
that	NN	O	O
paclitaxel	NN	O	O
and	NN	O	O
other	NN	O	O
drugs	NN	O	O
that	NN	O	O
disturb	NN	O	O
microtubule	NN	O	B-protein
function	NN	O	O
kill	NN	O	O
cells	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
through	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
Bcl-2	NN	O	B-protein
antagonizes	NN	O	O
drug-induced	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
calcineurin	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
blocking	NN	O	O
NFAT	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
,	NN	O	O
and	NN	O	O
preventing	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
can	NN	O	O
be	NN	O	O
overcome	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
partially	NN	O	O
,	NN	O	O
through	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

Phosphorylated	NN	O	O
Bcl-2	NN	O	B-protein
can	NN	O	O
not	NN	O	O
bind	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
can	NN	O	O
occur	NN	O	O
after	NN	O	O
Bcl-2	NN	O	B-protein
phosphorylation	NN	O	O
.	NN	O	O

-DOCSTART-	O

MHC-peptide	NN	O	O
ligand	NN	O	O
interactions	NN	O	O
establish	NN	O	O
a	NN	O	O
functional	NN	O	O
threshold	NN	O	O
for	NN	O	O
antigen-specific	NN	O	O
T	NN	O	O
cell	NN	O	O
recognition	NN	O	O
.	NN	O	O

Antigen-specific	NN	O	O
T	NN	O	O
cell	NN	O	O
recognition	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
functional	NN	O	O
density	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
-ligand	NN	O	O
,	NN	O	O
which	NN	O	O
consists	NN	O	O
of	NN	O	O
specific	NN	O	O
MHC	NN	O	B-protein
molecules	NN	O	I-protein
and	NN	O	O
a	NN	O	O
specifically	NN	O	O
bound	NN	O	O
peptide	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
the	NN	O	O
affinity	NN	O	O
and	NN	O	O
concentration	NN	O	O
of	NN	O	O
exogenous	NN	O	O
peptide	NN	O	O
and	NN	O	O
the	NN	O	O
density	NN	O	O
of	NN	O	O
specific	NN	O	O
MHC	NN	O	B-protein
molecules	NN	O	I-protein
on	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
a	NN	O	O
CD4+	NN	O	B-cell_line
,	NN	O	I-cell_line
DQA1*0501/DQB1*0201	NN	O	I-cell_line
(	NN	O	I-cell_line
DQ2.1	NN	O	I-cell_line
)	NN	O	I-cell_line
-restricted	NN	O	I-cell_line
,	NN	O	I-cell_line
HSV-2-specific	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
.	NN	O	O

Using	NN	O	O
antigen	NN	O	O
peptide	NN	O	O
analogs	NN	O	O
with	NN	O	O
different	NN	O	O
mutations	NN	O	O
of	NN	O	O
known	NN	O	O
DQ2-anchor	NN	O	O
residues	NN	O	O
,	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
was	NN	O	O
reduced	NN	O	O
in	NN	O	O
an	NN	O	O
peptide-affinity	NN	O	O
and	NN	O	O
-	NN	O	O
concentration	NN	O	O
specific	NN	O	O
manner	NN	O	O
.	NN	O	O

The	NN	O	O
decrease	NN	O	O
using	NN	O	O
weaker	NN	O	O
binding	NN	O	O
peptides	NN	O	O
was	NN	O	O
gradual	NN	O	O
as	NN	O	O
stimulation	NN	O	O
with	NN	O	O
a	NN	O	O
peptide	NN	O	O
with	NN	O	O
intermediate	NN	O	O
affinity	NN	O	O
yielded	NN	O	O
intermediate	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
and	NN	O	O
the	NN	O	O
poorest	NN	O	O
binding	NN	O	O
peptide	NN	O	O
induced	NN	O	O
an	NN	O	O
even	NN	O	O
weaker	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
response	NN	O	O
.	NN	O	O

MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
density	NN	O	O
on	NN	O	O
the	NN	O	O
APC	NN	O	B-cell_type
was	NN	O	O
modified	NN	O	O
using	NN	O	O
DQ2	NN	O	B-cell_line
homo	NN	O	I-cell_line
-and	NN	O	I-cell_line
heterozygous	NN	O	I-cell_line
B-LCLs	NN	O	I-cell_line
as	NN	O	O
APCs	NN	O	B-cell_type
,	NN	O	O
however	NN	O	O
this	NN	O	O
variation	NN	O	O
of	NN	O	O
MHC	NN	O	O
concentration	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
.	NN	O	O

We	NN	O	O
interpret	NN	O	O
this	NN	O	O
as	NN	O	O
a	NN	O	O
reflection	NN	O	O
of	NN	O	O
a	NN	O	O
low	NN	O	O
threshold	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
which	NN	O	O
peptide-	NN	O	O
MHC	NN	O	B-protein
avidity	NN	O	O
is	NN	O	O
the	NN	O	O
over-riding	NN	O	O
determinant	NN	O	O
of	NN	O	O
the	NN	O	O
strength	NN	O	O
of	NN	O	O
ligand	NN	O	O
signal	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulatory	NN	O	O
effects	NN	O	O
of	NN	O	O
interleukin-11	NN	O	B-protein
during	NN	O	O
acute	NN	O	O
lung	NN	O	O
inflammatory	NN	O	O
injury	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
interleukin-11	NN	O	B-protein
(	NN	O	O
IL-11	NN	O	B-protein
)	NN	O	O
was	NN	O	O
evaluated	NN	O	O
in	NN	O	O
the	NN	O	O
IgG	NN	O	B-protein
immune	NN	O	I-protein
complex	NN	O	I-protein
model	NN	O	O
of	NN	O	O
acute	NN	O	O
lung	NN	O	O
injury	NN	O	O
in	NN	O	O
rats	NN	O	O
.	NN	O	O

IL-11	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
were	NN	O	O
both	NN	O	O
up-regulated	NN	O	O
during	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
this	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
.	NN	O	O

Exogenously	NN	O	O
administered	NN	O	O
IL-11	NN	O	B-protein
substantially	NN	O	O
reduced	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
the	NN	O	O
intrapulmonary	NN	O	O
accumulation	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
the	NN	O	O
lung	NN	O	O
vascular	NN	O	O
leak	NN	O	O
of	NN	O	O
albumin	NN	O	B-protein
.	NN	O	O

These	NN	O	O
in	NN	O	O
vivo	NN	O	O
anti-inflammatory	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-11	NN	O	B-protein
were	NN	O	O
associated	NN	O	O
with	NN	O	O
reduced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
lung	NN	O	O
,	NN	O	O
reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
in	NN	O	O
bronchoalveolar	NN	O	O
lavage	NN	O	O
(	NN	O	O
BAL	NN	O	O
)	NN	O	O
fluids	NN	O	O
,	NN	O	O
and	NN	O	O
diminished	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
lung	NN	O	B-protein
vascular	NN	O	I-protein
ICAM-1	NN	O	I-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
interesting	NN	O	O
that	NN	O	O
IL-11	NN	O	B-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
BAL	NN	O	O
fluid	NN	O	O
content	NN	O	O
of	NN	O	O
the	NN	O	O
CXC	NN	O	B-protein
chemokines	NN	O	I-protein
,	NN	O	O
macrophage	NN	O	B-protein
inflammatory	NN	O	I-protein
protein-2	NN	O	I-protein
(	NN	O	O
MIP-2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
cytokine-inducible	NN	O	B-protein
neutrophil	NN	O	I-protein
chemoattractant	NN	O	I-protein
(	NN	O	O
CINC	NN	O	B-protein
)	NN	O	O
;	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-11	NN	O	B-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
these	NN	O	O
chemokines	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
BAL	NN	O	O
content	NN	O	O
of	NN	O	O
C5a	NN	O	O
was	NN	O	O
reduced	NN	O	O
by	NN	O	O
IL-11	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-11	NN	O	B-protein
is	NN	O	O
a	NN	O	O
regulatory	NN	O	B-protein
cytokine	NN	O	I-protein
in	NN	O	O
the	NN	O	O
lung	NN	O	O
and	NN	O	O
that	NN	O	O
,	NN	O	O
like	NN	O	O
other	NN	O	O
members	NN	O	O
of	NN	O	O
this	NN	O	O
family	NN	O	O
,	NN	O	O
its	NN	O	O
anti-inflammatory	NN	O	O
properties	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
linked	NN	O	O
to	NN	O	O
its	NN	O	O
suppression	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
diminished	NN	O	O
production	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
reduced	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
lung	NN	O	B-protein
vascular	NN	O	I-protein
ICAM-1	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Thrombopoietin	NN	O	B-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
and	NN	O	O
Stat5	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
platelets	NN	O	I-cell_type
.	NN	O	O

Thrombopoietin	NN	O	B-protein
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
essential	NN	O	O
for	NN	O	O
megakaryocytopoiesis	NN	O	O
and	NN	O	O
thrombopoiesis	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
and	NN	O	O
others	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
thrombopoietin	NN	O	B-protein
induces	NN	O	O
rapid	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak2	NN	O	B-protein
and	NN	O	O
other	NN	O	O
proteins	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
platelets	NN	O	I-cell_type
and	NN	O	O
BaF3	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
genetically	NN	O	O
engineered	NN	O	O
to	NN	O	O
express	NN	O	O
c-Mpl	NN	O	B-protein
,	NN	O	O
a	NN	O	O
receptor	NN	O	O
for	NN	O	O
thrombopoietin	NN	O	B-protein
.	NN	O	O

The	NN	O	O
Jak	NN	O	O
family	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
are	NN	O	O
known	NN	O	O
to	NN	O	O
mediate	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
or	NN	O	O
hematopoietic	NN	O	B-protein
growth	NN	O	I-protein
factors	NN	O	I-protein
by	NN	O	O
recruitment	NN	O	O
and	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
Stat	NN	O	B-protein
(	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
)	NN	O	O
proteins	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
whether	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
present	NN	O	O
in	NN	O	O
platelets	NN	O	B-cell_type
and	NN	O	O
,	NN	O	O
if	NN	O	O
so	NN	O	O
,	NN	O	O
whether	NN	O	O
they	NN	O	O
become	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
thrombopoietin	NN	O	B-protein
.	NN	O	O

We	NN	O	O
immunologically	NN	O	O
identified	NN	O	O
Stat1	NN	O	B-protein
,	NN	O	O
Stat2	NN	O	B-protein
,	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Stat5	NN	O	B-protein
in	NN	O	O
human	NN	O	O
platelet	NN	O	O
lysates	NN	O	O
.	NN	O	O

Thrombopoietin	NN	O	B-protein
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
and	NN	O	O
Stat5	NN	O	B-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Thrombopoietin	NN	O	B-protein
also	NN	O	O
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
and	NN	O	O
Stat5	NN	O	B-protein
in	NN	O	O
FDCP-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
genetically	NN	O	O
engineered	NN	O	O
to	NN	O	O
constitutively	NN	O	O
express	NN	O	O
human	NN	O	B-protein
c-Mpl	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
Stat3	NN	O	B-protein
and	NN	O	O
Stat5	NN	O	B-protein
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
after	NN	O	O
ligand	NN	O	O
binding	NN	O	O
to	NN	O	O
c-Mpl	NN	O	B-protein
and	NN	O	O
that	NN	O	O
this	NN	O	O
event	NN	O	O
may	NN	O	O
have	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
megakaryopoiesis/thrombopoiesis	NN	O	O
or	NN	O	O
possibly	NN	O	O
a	NN	O	O
mature	NN	O	O
platelet	NN	O	O
function	NN	O	O
such	NN	O	O
as	NN	O	O
aggregation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mechanisms	NN	O	O
that	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
lymphoid	NN	O	O
malignancies	NN	O	O
:	NN	O	O
roles	NN	O	O
for	NN	O	O
genetic	NN	O	O
alterations	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
defined	NN	O	O
genetic	NN	O	O
alterations	NN	O	O
commonly	NN	O	O
associated	NN	O	O
with	NN	O	O
transformed	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
review	NN	O	O
suggests	NN	O	O
roles	NN	O	O
for	NN	O	O
these	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
lymphoid	NN	O	O
neoplasms	NN	O	O
.	NN	O	O

Damage	NN	O	O
to	NN	O	O
the	NN	O	O
genes	NN	O	O
encoding	NN	O	O
proteins	NN	O	O
that	NN	O	O
function	NN	O	O
in	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
,	NN	O	O
transcription	NN	O	O
,	NN	O	O
or	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
and	NN	O	O
linked	NN	O	O
at	NN	O	O
varying	NN	O	O
degrees	NN	O	O
to	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
certain	NN	O	O
lymphoid	NN	O	O
malignancies	NN	O	O
.	NN	O	O

An	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
mechanistic	NN	O	O
consequences	NN	O	O
following	NN	O	O
such	NN	O	O
genetic	NN	O	O
alterations	NN	O	O
is	NN	O	O
essential	NN	O	O
to	NN	O	O
an	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
these	NN	O	O
lymphoid	NN	O	O
neoplasms	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
also	NN	O	O
becoming	NN	O	O
clear	NN	O	O
that	NN	O	O
the	NN	O	O
dysregulated	NN	O	O
expression	NN	O	O
of	NN	O	O
proteins	NN	O	O
that	NN	O	O
are	NN	O	O
not	NN	O	O
genetically	NN	O	O
altered	NN	O	O
can	NN	O	O
also	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
lymphoid	NN	O	O
malignancies	NN	O	O
.	NN	O	O

One	NN	O	O
such	NN	O	O
example	NN	O	O
is	NN	O	O
the	NN	O	O
excessive	NN	O	O
expression	NN	O	O
of	NN	O	O
``	NN	O	B-protein
normal	NN	O	I-protein
''	NN	O	I-protein
lymphokines	NN	O	I-protein
of	NN	O	O
cytokines	NN	O	B-protein
which	NN	O	O
accompanies	NN	O	O
many	NN	O	O
lymphoproliferative	NN	O	O
diseases	NN	O	O
.	NN	O	O

The	NN	O	O
dysregulated	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
during	NN	O	O
malignancy	NN	O	O
can	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
augmentation	NN	O	O
of	NN	O	O
growth	NN	O	O
of	NN	O	O
transformed	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
an	NN	O	O
alteration	NN	O	O
of	NN	O	O
the	NN	O	O
anti-tumor	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
latter	NN	O	O
mechanism	NN	O	O
is	NN	O	O
especially	NN	O	O
important	NN	O	O
because	NN	O	O
evasion	NN	O	O
of	NN	O	O
the	NN	O	O
impending	NN	O	O
immune	NN	O	O
response	NN	O	O
is	NN	O	O
a	NN	O	O
prerequisite	NN	O	O
for	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
lymphoproliferative	NN	O	O
diseases	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
this	NN	O	O
review	NN	O	O
supports	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
lymphoid	NN	O	O
malignancies	NN	O	O
is	NN	O	O
multifactorial	NN	O	O
,	NN	O	O
involving	NN	O	O
genetic	NN	O	O
alterations	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
dysregulated	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Dehydroepiandrosterone	NN	O	O
modulation	NN	O	O
of	NN	O	O
lipopolysaccharide-stimulated	NN	O	O
monocyte	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

Dehydroepiandrosterone	NN	O	O
(	NN	O	O
DHEA	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
predominant	NN	O	O
androgen	NN	O	O
secreted	NN	O	O
by	NN	O	O
the	NN	O	O
adrenal	NN	O	O
cortex	NN	O	O
,	NN	O	O
can	NN	O	O
be	NN	O	O
converted	NN	O	O
to	NN	O	O
both	NN	O	O
potent	NN	O	O
androgens	NN	O	O
and	NN	O	O
estrogens	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
its	NN	O	O
role	NN	O	O
as	NN	O	O
a	NN	O	O
precursor	NN	O	O
for	NN	O	O
other	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
,	NN	O	O
DHEA	NN	O	O
has	NN	O	O
been	NN	O	O
proposed	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
immunity	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
has	NN	O	O
investigated	NN	O	O
DHEA	NN	O	O
modulation	NN	O	O
of	NN	O	O
LPS-induced	NN	O	O
monocyte	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

Cytotoxicity	NN	O	B-protein
markers	NN	O	I-protein
assessed	NN	O	O
include	NN	O	O
tumor	NN	O	O
cell	NN	O	O
killing	NN	O	O
,	NN	O	O
IL-1	NN	O	B-protein
secretion	NN	O	O
,	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediate	NN	O	O
release	NN	O	O
,	NN	O	O
nitric	NN	O	B-protein
oxide	NN	O	I-protein
synthetase	NN	O	I-protein
activity	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
reactive	NN	O	O
nitrogen	NN	O	O
intermediates	NN	O	O
,	NN	O	O
complement	NN	O	B-protein
receptor-1	NN	O	I-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
protein	NN	O	I-protein
presence	NN	O	O
.	NN	O	O

Monocytes	NN	O	B-cell_type
stimulated	NN	O	O
with	NN	O	O
LPS	NN	O	O
concentrations	NN	O	O
of	NN	O	O
1.0	NN	O	O
micrograms/ml	NN	O	O
displayed	NN	O	O
the	NN	O	O
above	NN	O	O
cytotoxic	NN	O	O
markers	NN	O	O
,	NN	O	O
whereas	NN	O	O
monocytes	NN	O	B-cell_type
stimulated	NN	O	O
with	NN	O	O
DHEA	NN	O	O
alone	NN	O	O
or	NN	O	O
with	NN	O	O
LPS	NN	O	O
at	NN	O	O
a	NN	O	O
lower	NN	O	O
concentration	NN	O	O
of	NN	O	O
0.2	NN	O	O
ng/ml	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
when	NN	O	O
used	NN	O	O
simultaneously	NN	O	O
,	NN	O	O
DHEA	NN	O	O
and	NN	O	O
LPS	NN	O	O
0.2	NN	O	O
ng/ml	NN	O	O
displayed	NN	O	O
a	NN	O	O
synergistic	NN	O	O
effect	NN	O	O
on	NN	O	O
monocyte	NN	O	O
cytotoxicity	NN	O	O
against	NN	O	O
cancerous	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
IL-1	NN	O	B-protein
secretion	NN	O	O
,	NN	O	O
reactive	NN	O	O
nitrogen	NN	O	O
intermediate	NN	O	O
release	NN	O	O
,	NN	O	O
complement	NN	O	B-protein
receptor-1	NN	O	I-protein
cell-surface	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
protein	NN	O	I-protein
to	NN	O	O
levels	NN	O	O
comparable	NN	O	O
with	NN	O	O
levels	NN	O	O
obtained	NN	O	O
using	NN	O	O
LPS	NN	O	O
1.0	NN	O	O
microgram/ml	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
Scatchard	NN	O	O
plot	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
DHEA	NN	O	O
receptor	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
DHEA	NN	O	O
effects	NN	O	O
on	NN	O	O
LPS-stimulated	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
are	NN	O	O
mediated	NN	O	O
through	NN	O	O
a	NN	O	O
receptor-dependent	NN	O	O
process	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interferons	NN	O	O
inhibit	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
by	NN	O	O
interleukin	NN	O	B-protein
4	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
by	NN	O	O
inducing	NN	O	O
SOCS-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Interferons	NN	O	B-protein
(	NN	O	O
IFNs	NN	O	B-protein
)	NN	O	O
inhibit	NN	O	O
induction	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
of	NN	O	O
multiple	NN	O	O
genes	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
IFNs	NN	O	B-protein
mediate	NN	O	O
this	NN	O	O
inhibition	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
defined	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
activates	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
inducing	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
homodimerization	NN	O	O
,	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
latent	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
STAT6	NN	O	B-protein
(	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription-6	NN	O	I-protein
)	NN	O	O
.	NN	O	O

STAT6	NN	O	B-protein
-responsive	NN	O	O
elements	NN	O	O
are	NN	O	O
characteristically	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
IL-4-inducible	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Because	NN	O	O
STAT6	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
type	NN	O	O
I	NN	O	O
and	NN	O	O
type	NN	O	O
II	NN	O	O
IFNs	NN	O	B-protein
to	NN	O	O
regulate	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
by	NN	O	O
IL-4	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
with	NN	O	O
IFN-beta	NN	O	B-protein
or	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-1	NN	O	B-protein
,	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
granulocyte/macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
or	NN	O	O
transforming	NN	O	O
growth	NN	O	O
factor	NN	O	O
beta	NN	O	O
suppressed	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
by	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
decreased	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
and	NN	O	O
was	NN	O	O
not	NN	O	O
evident	NN	O	O
unless	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
preincubated	NN	O	O
with	NN	O	O
IFN	NN	O	B-protein
for	NN	O	O
at	NN	O	O
least	NN	O	O
1	NN	O	O
hr	NN	O	O
before	NN	O	O
IL-4	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
inhibition	NN	O	O
by	NN	O	O
IFN	NN	O	B-protein
could	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
cotreatment	NN	O	O
with	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
and	NN	O	O
correlated	NN	O	O
temporally	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
JAK/STAT	NN	O	B-DNA
inhibitory	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
SOCS-1	NN	O	B-DNA
.	NN	O	O

Forced	NN	O	O
expression	NN	O	O
of	NN	O	O
SOCS-1	NN	O	B-DNA
in	NN	O	O
a	NN	O	O
macrophage	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
RAW264	NN	O	B-cell_line
,	NN	O	O
markedly	NN	O	O
suppressed	NN	O	O
trans-activation	NN	O	O
of	NN	O	O
an	NN	O	O
IL-4-inducible	NN	O	B-DNA
reporter	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
IL-6-	NN	O	O
and	NN	O	O
IFN-gamma-induced	NN	O	O
reporter	NN	O	O
gene	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
IFNs	NN	O	B-protein
inhibit	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
and	NN	O	O
STAT6	NN	O	B-protein
-dependent	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
inducing	NN	O	O
expression	NN	O	O
of	NN	O	O
SOCS-1	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
activation-responsive	NN	O	O
element	NN	O	O
in	NN	O	O
single	NN	O	O
C	NN	O	B-DNA
motif-1/lymphotactin	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
site	NN	O	O
of	NN	O	O
constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
DNA-protein	NN	O	O
interactions	NN	O	O
involving	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
.	NN	O	O

Single	NN	O	B-protein
C	NN	O	I-protein
motif-1	NN	O	I-protein
(	NN	O	I-protein
SCM-1	NN	O	I-protein
)	NN	O	I-protein
/lymphotactin	NN	O	I-protein
is	NN	O	O
a	NN	O	O
C-type	NN	O	B-protein
chemokine	NN	O	I-protein
whose	NN	O	O
expression	NN	O	O
is	NN	O	O
activation	NN	O	O
dependent	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
sensitive	NN	O	O
and	NN	O	O
restricted	NN	O	O
to	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
double-negative	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
,	NN	O	O
gammadelta-type	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
humans	NN	O	O
,	NN	O	O
there	NN	O	O
are	NN	O	O
two	NN	O	O
highly	NN	O	O
homologous	NN	O	B-DNA
genes	NN	O	I-DNA
encoding	NN	O	O
SCM-1alpha	NN	O	B-protein
and	NN	O	O
SCM-1beta	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
regulatory	NN	O	O
mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
SCM-1	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
0.7	NN	O	O
kb	NN	O	O
was	NN	O	O
strongly	NN	O	O
induced	NN	O	O
upon	NN	O	O
activation	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
or	NN	O	O
PHA	NN	O	B-protein
plus	NN	O	O
PMA	NN	O	O
only	NN	O	O
in	NN	O	O
SCM-1-producer	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
through	NN	O	O
a	NN	O	O
cyclosporin	NN	O	O
A-sensitive	NN	O	O
mechanism	NN	O	O
.	NN	O	O

An	NN	O	O
element	NN	O	O
termed	NN	O	O
E1	NN	O	B-DNA
located	NN	O	O
at	NN	O	O
-108	NN	O	B-DNA
to	NN	O	I-DNA
-95	NN	O	I-DNA
nt	NN	O	I-DNA
relative	NN	O	O
to	NN	O	O
the	NN	O	O
major	NN	O	O
transcription	NN	O	O
start	NN	O	O
site	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
using	NN	O	O
the	NN	O	O
E1	NN	O	B-DNA
oligonucleotide	NN	O	O
as	NN	O	O
probe	NN	O	O
,	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
unstimulated	NN	O	O
T	NN	O	O
and	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
formed	NN	O	O
a	NN	O	O
constitutive	NN	O	B-protein
complex	NN	O	I-protein
termed	NN	O	O
complex	NN	O	B-protein
I	NN	O	I-protein
,	NN	O	O
while	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
stimulated	NN	O	O
SCM-1-producer	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
formed	NN	O	O
a	NN	O	O
higher	NN	O	B-protein
mobility	NN	O	I-protein
complex	NN	O	I-protein
termed	NN	O	O
complex	NN	O	B-protein
II	NN	O	I-protein
with	NN	O	O
a	NN	O	O
concomitant	NN	O	O
decrease	NN	O	O
in	NN	O	O
complex	NN	O	B-protein
I	NN	O	I-protein
.	NN	O	O

The	NN	O	O
shift	NN	O	O
from	NN	O	O
complex	NN	O	B-protein
I	NN	O	I-protein
to	NN	O	O
complex	NN	O	B-protein
II	NN	O	I-protein
seen	NN	O	O
only	NN	O	O
in	NN	O	O
SCM-1-producer	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
upon	NN	O	O
activation	NN	O	O
was	NN	O	O
completely	NN	O	O
suppressed	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

Both	NN	O	O
complexes	NN	O	O
were	NN	O	O
critically	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-DNA
core	NN	O	I-DNA
sequence	NN	O	I-DNA
TTTCC	NN	O	O
in	NN	O	O
the	NN	O	O
E1	NN	O	B-DNA
element	NN	O	I-DNA
and	NN	O	O
were	NN	O	O
partially	NN	O	O
supershifted	NN	O	O
by	NN	O	O
anti-NF-ATp	NN	O	B-protein
.	NN	O	O

One-hybrid	NN	O	O
assays	NN	O	O
in	NN	O	O
yeast	NN	O	B-protein
isolated	NN	O	I-protein
NF-ATp	NN	O	I-protein
as	NN	O	O
an	NN	O	O
E1	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
transfection	NN	O	O
of	NN	O	O
NF-ATp	NN	O	B-protein
into	NN	O	O
T	NN	O	B-cell_line
and	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
strongly	NN	O	O
enhanced	NN	O	O
the	NN	O	O
activation-dependent	NN	O	B-protein
SCM-1	NN	O	I-protein
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
a	NN	O	O
unique	NN	O	O
mechanism	NN	O	O
involving	NN	O	O
NF-ATp	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
regulate	NN	O	O
the	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
and	NN	O	O
activation-dependent	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
SCM-1	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Dopamine	NN	O	O
stimulates	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
via	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
reported	NN	O	O
that	NN	O	O
lymphocytes	NN	O	B-cell_type
produce	NN	O	O
,	NN	O	O
transport	NN	O	O
and	NN	O	O
bind	NN	O	O
dopamine	NN	O	O
present	NN	O	O
in	NN	O	O
plasma	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
dopamine	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
has	NN	O	O
not	NN	O	O
yet	NN	O	O
been	NN	O	O
examined	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
by	NN	O	O
dopamine	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

HIV-1	NN	O	O
replication	NN	O	O
was	NN	O	O
increased	NN	O	O
by	NN	O	O
dopamine	NN	O	O
,	NN	O	O
which	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
transactivation	NN	O	O
.	NN	O	O

Our	NN	O	O
transient	NN	O	O
expression	NN	O	O
data	NN	O	O
revealed	NN	O	O
that	NN	O	O
dopamine	NN	O	O
stimulated	NN	O	O
transcription	NN	O	O
through	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-DNA
element	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
importance	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-DNA
sites	NN	O	I-DNA
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
using	NN	O	O
vectors	NN	O	O
containing	NN	O	O
wild-type	NN	O	B-DNA
or	NN	O	I-DNA
mutant	NN	O	I-DNA
kappaB	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
mediating	NN	O	O
dopamine	NN	O	O
responsiveness	NN	O	O
,	NN	O	O
the	NN	O	O
proteasome	NN	O	O
inhibitor	NN	O	O
MG132	NN	O	O
abolished	NN	O	O
dopamine-induced	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
explored	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
dopamine	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
or	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
known	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
combination	NN	O	O
of	NN	O	O
dopamine	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
led	NN	O	O
to	NN	O	O
a	NN	O	O
stimulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
and	NN	O	O
replication	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
with	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
dopamine	NN	O	O
treatment	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
nor	NN	O	O
the	NN	O	O
concentrations	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
IkappaB-alpha	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
which	NN	O	O
suggests	NN	O	O
a	NN	O	O
distinct	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
mechanism	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
reveal	NN	O	O
a	NN	O	O
new	NN	O	O
link	NN	O	O
between	NN	O	O
the	NN	O	O
dopamine	NN	O	O
system	NN	O	O
,	NN	O	O
cytokine	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
via	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
PBMC	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Hormonal	NN	O	O
metabolic	NN	O	O
status	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
after	NN	O	O
conservative	NN	O	O
surgery	NN	O	O
:	NN	O	O
comparison	NN	O	O
with	NN	O	O
known	NN	O	O
prognostic	NN	O	O
criteria	NN	O	O
]	NN	O	O

Body	NN	O	O
weight	NN	O	O
,	NN	O	O
body	NN	O	O
mass	NN	O	O
index	NN	O	O
,	NN	O	O
body	NN	O	O
fat	NN	O	O
,	NN	O	O
lean	NN	O	O
body	NN	O	O
mass	NN	O	O
,	NN	O	O
blood-glucose	NN	O	O
,	NN	O	O
cholesterol	NN	O	O
,	NN	O	O
HDL-cholesterol	NN	O	B-protein
,	NN	O	O
triglyceride	NN	O	O
,	NN	O	O
beta-lipoproteins	NN	O	B-protein
,	NN	O	O
insulin	NN	O	O
,	NN	O	O
gonadotropin	NN	O	O
,	NN	O	O
estradiol	NN	O	O
,	NN	O	O
testosterone	NN	O	O
,	NN	O	O
SHBG	NN	O	B-protein
,	NN	O	O
T3	NN	O	B-protein
,	NN	O	O
T4	NN	O	B-protein
and	NN	O	O
TSH	NN	O	O
levels	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
estradiol	NN	O	O
and	NN	O	O
progesterone	NN	O	O
receptor	NN	O	O
levels	NN	O	O
in	NN	O	O
excised	NN	O	O
tumor	NN	O	O
were	NN	O	O
studied	NN	O	O
in	NN	O	O
40	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
prior	NN	O	O
to	NN	O	O
conservative	NN	O	O
treatment	NN	O	O
.	NN	O	O

Said	NN	O	O
anthropometric	NN	O	O
,	NN	O	O
metabolic	NN	O	O
and	NN	O	O
hormonal	NN	O	O
parameters	NN	O	O
were	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
index	NN	O	O
of	NN	O	O
lymphocytic	NN	O	O
infiltration	NN	O	O
of	NN	O	O
tumor	NN	O	O
selected	NN	O	O
as	NN	O	O
a	NN	O	O
prognostic	NN	O	O
factor	NN	O	O
.	NN	O	O

A	NN	O	O
significant	NN	O	O
correlation	NN	O	O
between	NN	O	O
high	NN	O	O
lymphocytic	NN	O	O
infiltration	NN	O	O
(	NN	O	O
2.5	NN	O	O
points	NN	O	O
)	NN	O	O
,	NN	O	O
low	NN	O	O
body	NN	O	O
mass	NN	O	O
and	NN	O	O
fat	NN	O	O
was	NN	O	O
identified	NN	O	O
.	NN	O	O

Also	NN	O	O
,	NN	O	O
smoking	NN	O	O
contributed	NN	O	O
to	NN	O	O
loss	NN	O	O
of	NN	O	O
body	NN	O	O
mass	NN	O	O
and	NN	O	O
fat	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
caused	NN	O	O
lymphocytic	NN	O	O
infiltration	NN	O	O
to	NN	O	O
rise	NN	O	O
.	NN	O	O

Moderate	NN	O	O
body	NN	O	O
mass	NN	O	O
,	NN	O	O
relatively	NN	O	O
low	NN	O	O
fat	NN	O	O
level	NN	O	O
and	NN	O	O
positive	NN	O	O
receptor	NN	O	O
status	NN	O	O
are	NN	O	O
among	NN	O	O
factors	NN	O	O
of	NN	O	O
good	NN	O	O
prognosis	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
of	NN	O	O
early	NN	O	O
stages	NN	O	O
.	NN	O	O

-DOCSTART-	O

Selective	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
interleukin-10-stimulated	NN	O	B-protein
STAT	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interleukin-10	NN	O	B-protein
(	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
have	NN	O	O
various	NN	O	O
reverse	NN	O	O
effects	NN	O	O
on	NN	O	O
macrophages	NN	O	B-cell_type
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
this	NN	O	O
difference	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
fully	NN	O	O
understood	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
IL-10-	NN	O	B-protein
and	NN	O	I-protein
IFN-gamma-activated	NN	O	I-protein
STAT	NN	O	I-protein
molecules	NN	O	I-protein
to	NN	O	O
two	NN	O	O
kinds	NN	O	O
of	NN	O	O
GAS	NN	O	B-DNA
-motif	NN	O	O
sequences	NN	O	O
.	NN	O	O

IL-10-activated	NN	O	B-protein
STAT1	NN	O	I-protein
could	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
GAS-motif	NN	O	B-DNA
sequence	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoter	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
Fcgamma	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
to	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
COX-2	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
whereas	NN	O	O
IFN-gamma-activated	NN	O	B-protein
STAT1	NN	O	I-protein
and	NN	O	O
STAT5	NN	O	B-protein
could	NN	O	O
bind	NN	O	O
to	NN	O	O
both	NN	O	O
sequences	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
inhibited	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
STAT	NN	O	B-protein
activation	NN	O	O
without	NN	O	O
newly	NN	O	B-protein
synthesized	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
aspirin	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
suppressed	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
STAT	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
IL-10-activated	NN	O	B-protein
STAT1	NN	O	I-protein
has	NN	O	O
a	NN	O	O
specificity	NN	O	O
in	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
GAS-motif	NN	O	B-DNA
sequences	NN	O	I-DNA
,	NN	O	O
whereas	NN	O	O
IFN-gamma-activated	NN	O	B-protein
STAT1	NN	O	I-protein
and	NN	O	O
STAT5	NN	O	B-protein
have	NN	O	O
a	NN	O	O
broader	NN	O	O
spectrum	NN	O	O
in	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
GAS-motif	NN	O	B-DNA
sequences	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
may	NN	O	O
explain	NN	O	O
the	NN	O	O
difference	NN	O	O
between	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
biological	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
on	NN	O	O
IFN-gamma	NN	O	B-protein
activities	NN	O	O
.	NN	O	O

-DOCSTART-	O

Peptide	NN	O	O
binding	NN	O	O
affinity	NN	O	O
and	NN	O	O
pH	NN	O	O
variation	NN	O	O
establish	NN	O	O
functional	NN	O	O
thresholds	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
HLA-DQ	NN	O	B-DNA
-restricted	NN	O	O
T	NN	O	O
cell	NN	O	O
recognition	NN	O	O
.	NN	O	O

Peptides	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
HSV-2	NN	O	B-protein
VP16	NN	O	I-protein
protein	NN	O	I-protein
were	NN	O	O
utilized	NN	O	O
for	NN	O	O
studies	NN	O	O
of	NN	O	O
peptide	NN	O	O
binding	NN	O	O
to	NN	O	O
DQ0302	NN	O	O
molecules	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
at	NN	O	O
both	NN	O	O
neutral	NN	O	O
and	NN	O	O
acidic	NN	O	O
pH	NN	O	O
.	NN	O	O

The	NN	O	O
native	NN	O	O
peptide	NN	O	O
VP16	NN	O	O
430-444	NN	O	O
contains	NN	O	O
an	NN	O	O
Asp	NN	O	O
at	NN	O	O
position	NN	O	B-protein
442	NN	O	I-protein
,	NN	O	O
binds	NN	O	O
to	NN	O	O
DQ0302	NN	O	O
strongly	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
Kd	NN	O	O
value	NN	O	O
of	NN	O	O
50nM	NN	O	O
at	NN	O	O
acidic	NN	O	O
pH	NN	O	O
and	NN	O	O
very	NN	O	O
weakly	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
Kd	NN	O	O
value	NN	O	O
of	NN	O	O
greater	NN	O	O
than	NN	O	O
10	NN	O	O
microM	NN	O	O
at	NN	O	O
neutral	NN	O	O
pH	NN	O	O
.	NN	O	O

A	NN	O	O
truncated	NN	O	O
version	NN	O	O
of	NN	O	O
430-444	NN	O	O
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
VP16	NN	O	O
433-442	NN	O	O
,	NN	O	O
binds	NN	O	O
with	NN	O	O
an	NN	O	O
affinity	NN	O	O
10-fold	NN	O	O
lower	NN	O	O
compared	NN	O	O
to	NN	O	O
430-444	NN	O	O
at	NN	O	O
acidic	NN	O	O
pH	NN	O	O
,	NN	O	O
and	NN	O	O
binding	NN	O	O
at	NN	O	O
neutral	NN	O	O
pH	NN	O	O
was	NN	O	O
barely	NN	O	O
detectable	NN	O	O
.	NN	O	O

The	NN	O	O
homologous	NN	O	O
peptide	NN	O	O
430-444	NN	O	O
,	NN	O	O
442A	NN	O	O
has	NN	O	O
an	NN	O	O
Asp	NN	O	O
to	NN	O	O
Ala	NN	O	O
substitution	NN	O	O
at	NN	O	O
position	NN	O	O
442	NN	O	O
and	NN	O	O
binds	NN	O	O
to	NN	O	O
DQ0302	NN	O	O
with	NN	O	O
a	NN	O	O
Kd	NN	O	O
similar	NN	O	O
to	NN	O	O
433-442	NN	O	O
.	NN	O	O

The	NN	O	O
short	NN	O	O
truncated	NN	O	O
analog	NN	O	O
433-442A	NN	O	O
binds	NN	O	O
very	NN	O	O
poorly	NN	O	O
at	NN	O	O
both	NN	O	O
acidic	NN	O	O
and	NN	O	O
neutral	NN	O	O
pH	NN	O	O
.	NN	O	O

Both	NN	O	O
the	NN	O	O
wild	NN	O	O
type	NN	O	O
430-444	NN	O	O
and	NN	O	O
433-442	NN	O	O
peptides	NN	O	O
stimulated	NN	O	O
a	NN	O	O
HSV-specific	NN	O	O
T	NN	O	O
cell	NN	O	O
clone	NN	O	O
after	NN	O	O
a	NN	O	O
brief	NN	O	O
incubation	NN	O	O
with	NN	O	O
antigen	NN	O	B-cell_type
presenting	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
APC	NN	O	B-cell_type
)	NN	O	O
expressing	NN	O	O
DQ0302	NN	O	O
at	NN	O	O
acidic	NN	O	O
pH	NN	O	O
.	NN	O	O

Much	NN	O	O
higher	NN	O	O
concentrations	NN	O	O
of	NN	O	O
wild	NN	O	O
type	NN	O	O
peptides	NN	O	O
were	NN	O	O
needed	NN	O	O
to	NN	O	O
activate	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
neutral	NN	O	O
pH	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
APC	NN	O	B-cell_type
pulsed	NN	O	O
with	NN	O	O
Ala-substituted	NN	O	O
peptides	NN	O	O
430-444	NN	O	O
,	NN	O	O
442A	NN	O	O
or	NN	O	O
433-442A	NN	O	O
at	NN	O	O
neutral	NN	O	O
pH	NN	O	O
failed	NN	O	O
to	NN	O	O
stimulate	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
APC	NN	O	B-cell_type
pulsed	NN	O	O
at	NN	O	O
acidic	NN	O	O
pH	NN	O	O
and	NN	O	O
subsequently	NN	O	O
washed	NN	O	O
led	NN	O	O
to	NN	O	O
successful	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
Ala-substituted	NN	O	O
peptide	NN	O	O
was	NN	O	O
recognized	NN	O	O
by	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
at	NN	O	O
neutral	NN	O	O
pH	NN	O	O
only	NN	O	O
when	NN	O	O
it	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
APC	NN	O	B-cell_line
culture	NN	O	I-cell_line
throughout	NN	O	O
the	NN	O	O
stimulation	NN	O	O
process	NN	O	O
.	NN	O	O

While	NN	O	O
the	NN	O	O
MHC-peptide	NN	O	B-protein
complexes	NN	O	I-protein
formed	NN	O	O
with	NN	O	O
the	NN	O	O
native	NN	O	O
peptide	NN	O	O
are	NN	O	O
stable	NN	O	O
,	NN	O	O
complexes	NN	O	O
formed	NN	O	O
with	NN	O	O
the	NN	O	O
Ala-substituted	NN	O	O
peptide	NN	O	O
had	NN	O	O
a	NN	O	O
functional	NN	O	O
t1/2	NN	O	O
of	NN	O	O
less	NN	O	O
than	NN	O	O
4	NN	O	O
hr	NN	O	O
at	NN	O	O
neutral	NN	O	O
pH	NN	O	O
.	NN	O	O

-DOCSTART-	O

ETS	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
regulate	NN	O	O
an	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
third	NN	O	B-DNA
intron	NN	O	I-DNA
of	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

We	NN	O	O
describe	NN	O	O
an	NN	O	O
enhancer	NN	O	O
site	NN	O	O
in	NN	O	O
the	NN	O	O
third	NN	O	B-DNA
intron	NN	O	I-DNA
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

A	NN	O	O
reporter	NN	O	B-DNA
construct	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
TNF-alpha	NN	O	B-DNA
gene	NN	O	I-DNA
displayed	NN	O	O
weak	NN	O	O
activity	NN	O	O
when	NN	O	O
transfected	NN	O	O
into	NN	O	O
RAW264.7	NN	O	B-cell_line
macrophage-like	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
the	NN	O	O
third	NN	O	B-DNA
intron	NN	O	I-DNA
of	NN	O	O
TNF-alpha	NN	O	B-protein
to	NN	O	O
this	NN	O	O
construct	NN	O	O
resulted	NN	O	O
in	NN	O	O
an	NN	O	O
enhancement	NN	O	O
of	NN	O	O
CAT	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

This	NN	O	O
enhancement	NN	O	O
was	NN	O	O
eliminated	NN	O	O
if	NN	O	O
a	NN	O	O
conserved	NN	O	B-DNA
20-bp	NN	O	I-DNA
sequence	NN	O	I-DNA
was	NN	O	O
removed	NN	O	O
from	NN	O	O
the	NN	O	O
intron	NN	O	B-DNA
or	NN	O	O
if	NN	O	O
a	NN	O	O
dominant-negative	NN	O	B-protein
ets-binding	NN	O	I-protein
factor	NN	O	I-protein
was	NN	O	O
co-transfected	NN	O	O
with	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Mutations	NN	O	O
of	NN	O	O
this	NN	O	O
site	NN	O	O
that	NN	O	O
destroyed	NN	O	O
potential	NN	O	O
ets	NN	O	B-DNA
transcription	NN	O	I-DNA
factor	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
had	NN	O	O
reduced	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
major	NN	O	O
transcription	NN	O	O
factor	NN	O	O
that	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
oligonucleotide	NN	O	O
was	NN	O	O
confirmed	NN	O	O
to	NN	O	O
be	NN	O	O
GABP	NN	O	B-protein
by	NN	O	O
supershift	NN	O	O
and	NN	O	O
competition	NN	O	O
analysis	NN	O	O
.	NN	O	O

In	NN	O	O
RAW264.7	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
binding	NN	O	O
was	NN	O	O
constitutive	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
in	NN	O	O
bone	NN	O	B-cell_line
marrow-derived	NN	O	I-cell_line
macrophages	NN	O	I-cell_line
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
interferon-gamma	NN	O	B-protein
inducible	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
imply	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
ets	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

E2F-1	NN	O	B-protein
blocks	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
and	NN	O	O
causes	NN	O	O
proliferation	NN	O	O
in	NN	O	O
transgenic	NN	O	B-cell_type
megakaryocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
E2F-1	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
through	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
activate	NN	O	B-DNA
genes	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
cell	NN	O	O
division	NN	O	O
.	NN	O	O

E2F-1	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
proteins	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	O
susceptibility	NN	O	B-DNA
gene	NN	O	I-DNA
product	NN	O	O
,	NN	O	O
cyclin-dependent	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
and	NN	O	O
their	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
proteins	NN	O	O
that	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
certain	NN	O	O
developmental	NN	O	O
processes	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
a	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
E2F-1	NN	O	B-protein
in	NN	O	O
differentiation	NN	O	O
,	NN	O	O
we	NN	O	O
assayed	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
megakaryocytes	NN	O	B-cell_type
to	NN	O	O
form	NN	O	O
platelets	NN	O	B-cell_type
in	NN	O	O
an	NN	O	O
in	NN	O	O
vivo	NN	O	O
transgenic	NN	O	O
model	NN	O	O
.	NN	O	O

E2F-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
megakaryocytes	NN	O	B-cell_type
blocked	NN	O	O
differentiation	NN	O	O
during	NN	O	O
maturation	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
severe	NN	O	O
thrombocytopenia	NN	O	O
.	NN	O	O

Ultrastructural	NN	O	O
analysis	NN	O	O
of	NN	O	O
megakaryocytes	NN	O	B-cell_type
revealed	NN	O	O
abnormal	NN	O	O
development	NN	O	O
characterized	NN	O	O
by	NN	O	O
hyperdemarcation	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	O
membranes	NN	O	O
and	NN	O	O
reduced	NN	O	O
numbers	NN	O	O
of	NN	O	O
alpha	NN	O	O
granules	NN	O	O
.	NN	O	O

Administration	NN	O	O
of	NN	O	O
megakaryocyte	NN	O	B-protein
growth	NN	O	I-protein
and	NN	O	I-protein
development	NN	O	I-protein
factor	NN	O	I-protein
or	NN	O	O
interleukin	NN	O	B-protein
6	NN	O	I-protein
could	NN	O	O
not	NN	O	O
overcome	NN	O	O
the	NN	O	O
differentiation	NN	O	O
block	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
E2F-1	NN	O	B-protein
caused	NN	O	O
massive	NN	O	O
megakaryocyte	NN	O	B-cell_type
accumulation	NN	O	O
in	NN	O	O
both	NN	O	O
normal	NN	O	O
and	NN	O	O
ectopic	NN	O	O
sites	NN	O	O
,	NN	O	O
first	NN	O	O
evident	NN	O	O
in	NN	O	O
E15	NN	O	O
embryonic	NN	O	O
liver	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
significant	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
transgenic	NN	O	B-cell_type
megakaryocytes	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
E2F-1	NN	O	B-protein
can	NN	O	O
prevent	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
,	NN	O	O
probably	NN	O	O
through	NN	O	O
its	NN	O	O
cell	NN	O	O
cycle-stimulatory	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Polymorphic	NN	O	O
nucleotides	NN	O	O
within	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
that	NN	O	O
mediate	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

Atopy	NN	O	O
,	NN	O	O
which	NN	O	O
predisposes	NN	O	O
individuals	NN	O	O
to	NN	O	O
develop	NN	O	O
asthma	NN	O	O
,	NN	O	O
severe	NN	O	O
systemic	NN	O	O
anaphylaxis	NN	O	O
,	NN	O	O
and	NN	O	O
atopic	NN	O	O
dermatitis	NN	O	O
,	NN	O	O
is	NN	O	O
usually	NN	O	O
associated	NN	O	O
with	NN	O	O
dramatically	NN	O	O
elevated	NN	O	O
total	NN	O	O
serum	NN	O	O
IgE	NN	O	B-protein
levels	NN	O	O
and	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
major	NN	O	O
susceptibility	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
multiple	NN	O	O
minor	NN	O	O
susceptibility	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
recent	NN	O	O
sib-pair	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
a	NN	O	O
tight	NN	O	O
linkage	NN	O	O
between	NN	O	O
markers	NN	O	O
on	NN	O	O
5q31.1	NN	O	B-DNA
and	NN	O	O
a	NN	O	O
major	NN	O	B-DNA
susceptibility	NN	O	I-DNA
gene	NN	O	I-DNA
controlling	NN	O	O
total	NN	O	O
serum	NN	O	O
IgE	NN	O	B-protein
levels	NN	O	O
.	NN	O	O

Due	NN	O	O
to	NN	O	O
its	NN	O	O
location	NN	O	O
within	NN	O	O
this	NN	O	O
cluster	NN	O	O
and	NN	O	O
its	NN	O	O
biologic	NN	O	O
role	NN	O	O
in	NN	O	O
Ig	NN	O	O
class	NN	O	O
switching	NN	O	O
and	NN	O	O
Th2	NN	O	B-cell_line
cell	NN	O	I-cell_line
differentiation	NN	O	O
,	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
has	NN	O	O
emerged	NN	O	O
as	NN	O	O
one	NN	O	O
major	NN	O	O
candidate	NN	O	O
for	NN	O	O
the	NN	O	O
atopy	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
one	NN	O	O
model	NN	O	O
,	NN	O	O
polymorphisms	NN	O	O
within	NN	O	O
IL-4	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
might	NN	O	O
result	NN	O	O
in	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
amplifying	NN	O	O
Th2	NN	O	B-cell_line
cell	NN	O	I-cell_line
differentiation	NN	O	O
and	NN	O	O
class	NN	O	O
switching	NN	O	O
to	NN	O	O
IgE	NN	O	B-protein
.	NN	O	O

In	NN	O	O
support	NN	O	O
of	NN	O	O
this	NN	O	O
model	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
exists	NN	O	O
in	NN	O	O
multiple	NN	O	O
allelic	NN	O	O
forms	NN	O	O
that	NN	O	O
exhibit	NN	O	O
distinct	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
in	NN	O	O
IL-4-positive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
particular	NN	O	O
allele	NN	O	O
has	NN	O	O
an	NN	O	O
unusually	NN	O	O
high	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

A	NN	O	O
nucleotide	NN	O	O
substitution	NN	O	O
within	NN	O	O
a	NN	O	O
recently	NN	O	O
described	NN	O	O
OAP40	NN	O	B-DNA
element	NN	O	I-DNA
located	NN	O	O
just	NN	O	O
upstream	NN	O	O
of	NN	O	O
an	NN	O	O
NF-AT	NN	O	B-DNA
site	NN	O	I-DNA
(	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
)	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
largely	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
increased	NN	O	O
promotor	NN	O	O
strength	NN	O	O
of	NN	O	O
this	NN	O	O
particular	NN	O	O
allelic	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
EMSAs	NN	O	O
,	NN	O	O
this	NN	O	O
substitution	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
markedly	NN	O	O
enhanced	NN	O	O
affinity	NN	O	O
for	NN	O	O
sequence-specific	NN	O	O
complexes	NN	O	O
exhibiting	NN	O	O
an	NN	O	O
AP-1	NN	O	B-protein
specificity	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
allelic	NN	O	O
nucleotides	NN	O	O
,	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
provides	NN	O	O
specific	NN	O	O
targets	NN	O	O
for	NN	O	O
a	NN	O	O
comprehensive	NN	O	O
screening	NN	O	O
of	NN	O	O
atopic	NN	O	O
and	NN	O	O
nonatopic	NN	O	O
individuals	NN	O	O
and	NN	O	O
may	NN	O	O
provide	NN	O	O
a	NN	O	O
clue	NN	O	O
for	NN	O	O
genetic	NN	O	O
predisposition	NN	O	O
for	NN	O	O
atopy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Evidence	NN	O	O
for	NN	O	O
lowered	NN	O	O
induction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
during	NN	O	O
aging	NN	O	O
.	NN	O	O

Transcription	NN	O	O
factor	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
is	NN	O	O
induced	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
young	NN	O	O
individuals	NN	O	O
following	NN	O	O
activation	NN	O	O
with	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
stimuli	NN	O	O
including	NN	O	O
anti-CD3	NN	O	B-protein
,	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
older	NN	O	O
individuals	NN	O	O
show	NN	O	O
a	NN	O	O
significant	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
stimuli	NN	O	O
.	NN	O	O

The	NN	O	O
age-related	NN	O	O
decline	NN	O	O
in	NN	O	O
induction	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
could	NN	O	O
not	NN	O	O
be	NN	O	O
attributed	NN	O	O
to	NN	O	O
alteration	NN	O	O
in	NN	O	O
the	NN	O	O
composition	NN	O	O
of	NN	O	O
subunits	NN	O	O
,	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
predominant	NN	O	O
subunits	NN	O	O
of	NN	O	O
induced	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
young	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
elderly	NN	O	O
donors	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
similar	NN	O	O
levels	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
cytosols	NN	O	O
of	NN	O	O
unactivated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
both	NN	O	O
young	NN	O	O
and	NN	O	O
elderly	NN	O	O
donors	NN	O	O
suggesting	NN	O	O
that	NN	O	O
precursor	NN	O	O
levels	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
remain	NN	O	O
unaltered	NN	O	O
during	NN	O	O
aging	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
an	NN	O	O
age-associated	NN	O	O
decline	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
elderly	NN	O	O
individuals	NN	O	O
may	NN	O	O
be	NN	O	O
attributable	NN	O	O
to	NN	O	O
altered	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
immune	NN	O	O
dysregulation	NN	O	O
accompanying	NN	O	O
aging	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
lipopolysaccharide-induced	NN	O	O
monocyte	NN	O	B-protein
interleukin-1	NN	O	I-protein
receptor	NN	O	I-protein
antagonist	NN	O	O
synthesis	NN	O	O
by	NN	O	O
cortisol	NN	O	O
:	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
mineralocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Glucocorticoids	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
part	NN	O	O
of	NN	O	O
their	NN	O	O
physiological	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
and	NN	O	O
immune	NN	O	O
processes	NN	O	O
,	NN	O	O
suppress	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

Human	NN	O	B-RNA
monocyte	NN	O	I-RNA
IL-1	NN	O	I-RNA
receptor	NN	O	I-RNA
antagonist	NN	O	I-RNA
(	NN	O	I-RNA
IL-1ra	NN	O	I-RNA
)	NN	O	I-RNA
messenger	NN	O	I-RNA
ribonucleic	NN	O	I-RNA
acid	NN	O	I-RNA
(	NN	O	O
mRNA	NN	O	O
)	NN	O	O
expression	NN	O	O
and	NN	O	O
protein	NN	O	O
secretion	NN	O	O
are	NN	O	O
inhibited	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
further	NN	O	O
studied	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-1ra	NN	O	B-protein
by	NN	O	O
the	NN	O	O
major	NN	O	O
physiological	NN	O	O
human	NN	O	O
glucocorticoid	NN	O	O
,	NN	O	O
cortisol	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
cortisol	NN	O	O
incubation	NN	O	O
induced	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
IL-1ra	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
a	NN	O	O
significant	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-1ra	NN	O	B-protein
protein	NN	O	I-protein
secretion	NN	O	O
in	NN	O	O
cell	NN	O	O
cultures	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
the	NN	O	O
bacterial	NN	O	O
endotoxin	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

Oral	NN	O	O
administration	NN	O	O
of	NN	O	O
276	NN	O	O
mumol	NN	O	O
cortisol	NN	O	O
to	NN	O	O
normal	NN	O	O
subjects	NN	O	O
also	NN	O	O
decreased	NN	O	O
LPS-induced	NN	O	O
IL-1ra	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
.	NN	O	O

By	NN	O	O
coincubating	NN	O	O
the	NN	O	O
monocytes	NN	O	B-cell_type
with	NN	O	O
either	NN	O	O
the	NN	O	O
mineralocorticoid	NN	O	O
antagonist	NN	O	O
spironolactone	NN	O	O
or	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
antagonist	NN	O	O
RU	NN	O	O
38486	NN	O	O
,	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
cortisol-induced	NN	O	O
inhibition	NN	O	O
of	NN	O	O
LPS-stimulated	NN	O	O
IL-1ra	NN	O	B-protein
secretion	NN	O	O
was	NN	O	O
partially	NN	O	O
reversed	NN	O	O
.	NN	O	O

The	NN	O	O
mineralocorticoid	NN	O	O
aldosterone	NN	O	O
exerted	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
in	NN	O	O
LPS-induced	NN	O	O
monocyte	NN	O	O
IL-1ra	NN	O	B-protein
secretion	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
coincubation	NN	O	O
with	NN	O	O
spironolactone	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
mineralocorticoid	NN	O	B-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
human	NN	O	O
monocytes	NN	O	B-cell_type
was	NN	O	O
observed	NN	O	O
by	NN	O	O
PCR	NN	O	O
of	NN	O	O
reversed	NN	O	B-RNA
transcribed	NN	O	I-RNA
RNA	NN	O	I-RNA
.	NN	O	O

Our	NN	O	O
results	NN	O	O
further	NN	O	O
indicate	NN	O	O
that	NN	O	O
corticosteroids	NN	O	O
physiologically	NN	O	O
control	NN	O	O
the	NN	O	O
IL-1/	NN	O	O
IL-1ra	NN	O	B-protein
system	NN	O	O
during	NN	O	O
inflammatory	NN	O	O
or	NN	O	O
immune	NN	O	O
processes	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
a	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
-mediated	NN	O	O
effect	NN	O	O
,	NN	O	O
the	NN	O	O
mineralocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
monocyte	NN	O	O
IL-1ra	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
cortisol	NN	O	O
.	NN	O	O

-DOCSTART-	O

Target	NN	O	O
structures	NN	O	O
of	NN	O	O
the	NN	O	O
CD8	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-T-cell	NN	O	O
response	NN	O	O
to	NN	O	O
human	NN	O	O
cytomegalovirus	NN	O	O
:	NN	O	O
the	NN	O	O
72-kilodalton	NN	O	B-protein
major	NN	O	I-protein
immediate-early	NN	O	I-protein
protein	NN	O	I-protein
revisited	NN	O	O
.	NN	O	O

Cell-mediated	NN	O	O
immunity	NN	O	O
plays	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
infection	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	O
cytomegalovirus	NN	O	O
(	NN	O	O
HCMV	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
only	NN	O	O
a	NN	O	O
few	NN	O	O
CD8	NN	O	B-protein
(	NN	O	I-protein
+	NN	O	I-protein
)	NN	O	I-protein
-T-cell	NN	O	I-protein
epitopes	NN	O	I-protein
are	NN	O	O
known	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
majority	NN	O	O
being	NN	O	O
contained	NN	O	O
in	NN	O	O
the	NN	O	O
pp65	NN	O	B-protein
phosphoprotein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
dominate	NN	O	O
the	NN	O	O
CD8	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-T-cell	NN	O	O
response	NN	O	O
to	NN	O	O
HCMV	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
readdressed	NN	O	O
the	NN	O	O
issue	NN	O	O
of	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
specific	NN	O	O
for	NN	O	O
the	NN	O	O
72-kDa	NN	O	B-protein
major	NN	O	I-protein
immediate-early	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
IE-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
nonstructural	NN	O	O
but	NN	O	O
is	NN	O	O
found	NN	O	O
very	NN	O	O
early	NN	O	O
and	NN	O	O
throughout	NN	O	O
the	NN	O	O
replicative	NN	O	O
cycle	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
novel	NN	O	O
flow-cytometric	NN	O	O
assay	NN	O	O
,	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
identify	NN	O	O
CD8	NN	O	B-protein
(	NN	O	I-protein
+	NN	O	I-protein
)	NN	O	I-protein
-T-cell	NN	O	I-protein
epitopes	NN	O	I-protein
(	NN	O	O
by	NN	O	O
IE-1	NN	O	B-protein
peptide-specific	NN	O	O
induction	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
synthesis	NN	O	O
)	NN	O	O
and	NN	O	O
simultaneously	NN	O	O
measure	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
cells	NN	O	O
directed	NN	O	O
against	NN	O	O
them	NN	O	O
.	NN	O	O

For	NN	O	O
this	NN	O	O
purpose	NN	O	O
,	NN	O	O
81	NN	O	O
pentadecamer	NN	O	O
peptides	NN	O	O
covering	NN	O	O
the	NN	O	O
complete	NN	O	O
491-amino-acid	NN	O	B-protein
sequence	NN	O	I-protein
of	NN	O	O
IE-1	NN	O	B-protein
were	NN	O	O
tested	NN	O	O
on	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
anti-HCMV	NN	O	O
immunoglobulin	NN	O	O
G-seropositive	NN	O	O
donors	NN	O	O
.	NN	O	O

At	NN	O	O
least	NN	O	O
10	NN	O	O
new	NN	O	O
epitopes	NN	O	B-protein
were	NN	O	O
identified	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
fine	NN	O	O
specificity	NN	O	O
and	NN	O	O
presenting	NN	O	O
HLA	NN	O	B-protein
molecule	NN	O	I-protein
of	NN	O	O
the	NN	O	O
first	NN	O	O
of	NN	O	O
them	NN	O	O
was	NN	O	O
determined	NN	O	O
.	NN	O	O

The	NN	O	O
frequencies	NN	O	O
of	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
directed	NN	O	O
against	NN	O	O
IE-1	NN	O	B-protein
were	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
directed	NN	O	O
against	NN	O	O
pp65	NN	O	B-protein
in	NN	O	O
donors	NN	O	O
tested	NN	O	O
with	NN	O	O
known	NN	O	O
pp65	NN	O	B-protein
-derived	NN	O	O
peptides	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
additional	NN	O	O
testing	NN	O	O
of	NN	O	O
a	NN	O	O
corresponding	NN	O	O
set	NN	O	O
of	NN	O	O
peptides	NN	O	O
covering	NN	O	O
the	NN	O	O
complete	NN	O	O
sequence	NN	O	O
of	NN	O	O
pp65	NN	O	B-protein
on	NN	O	O
10	NN	O	O
of	NN	O	O
these	NN	O	O
donors	NN	O	O
identified	NN	O	O
individuals	NN	O	O
whose	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
recognized	NN	O	O
IE-1	NN	O	B-protein
but	NN	O	O
not	NN	O	O
pp65	NN	O	B-protein
and	NN	O	O
vice	NN	O	O
versa	NN	O	O
,	NN	O	O
clearly	NN	O	O
illustrating	NN	O	O
that	NN	O	O
either	NN	O	O
protein	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
major	NN	O	O
target	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
IE-1	NN	O	B-protein
is	NN	O	O
far	NN	O	O
more	NN	O	O
important	NN	O	O
as	NN	O	O
a	NN	O	O
CD8	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-T-cell	NN	O	O
target	NN	O	O
than	NN	O	O
current	NN	O	O
opinion	NN	O	O
suggests	NN	O	O
.	NN	O	O

-DOCSTART-	O

E1A	NN	O	B-DNA
oncogene	NN	O	I-DNA
induction	NN	O	O
of	NN	O	O
cellular	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
killing	NN	O	O
by	NN	O	O
cytolytic	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
through	NN	O	O
target	NN	O	O
cell	NN	O	O
sensitization	NN	O	O
to	NN	O	O
apoptotic	NN	O	O
injury	NN	O	O
.	NN	O	O

E1A	NN	O	B-DNA
oncogene	NN	O	I-DNA
expression	NN	O	O
increases	NN	O	O
mammalian	NN	O	O
cell	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
lysis	NN	O	O
by	NN	O	O
cytolytic	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
CLs	NN	O	B-cell_type
)	NN	O	O
at	NN	O	O
a	NN	O	O
stage	NN	O	O
in	NN	O	O
this	NN	O	O
intercellular	NN	O	O
interaction	NN	O	O
that	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
cell	NN	O	O
surface	NN	O	O
recognition	NN	O	O
events	NN	O	O
.	NN	O	O

Since	NN	O	O
CLs	NN	O	B-cell_type
can	NN	O	O
induce	NN	O	O
either	NN	O	O
apoptotic	NN	O	O
or	NN	O	O
necrotic	NN	O	O
cell	NN	O	O
death	NN	O	O
,	NN	O	O
we	NN	O	O
asked	NN	O	O
whether	NN	O	O
E1A	NN	O	B-protein
sensitization	NN	O	O
to	NN	O	O
injury-induced	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
explain	NN	O	O
E1A	NN	O	B-protein
-induced	NN	O	O
cytolytic	NN	O	O
susceptibility	NN	O	O
.	NN	O	O

Mouse	NN	O	B-cell_type
,	NN	O	I-cell_type
rat	NN	O	I-cell_type
,	NN	O	I-cell_type
hamster	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
human	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
were	NN	O	O
rendered	NN	O	O
cytolytic	NN	O	O
susceptible	NN	O	O
by	NN	O	O
E1A	NN	O	B-protein
were	NN	O	O
also	NN	O	O
sensitized	NN	O	O
to	NN	O	O
CL-induced	NN	O	O
and	NN	O	O
chemically	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
E1A-positive	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
no	NN	O	O
more	NN	O	O
susceptible	NN	O	O
to	NN	O	O
injury-induced	NN	O	O
necrosis	NN	O	O
than	NN	O	O
E1A-negative	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
induction	NN	O	O
of	NN	O	O
cytolytic	NN	O	O
susceptibility	NN	O	O
and	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
other	NN	O	O
E1A	NN	O	B-protein
activities	NN	O	O
,	NN	O	O
cellular	NN	O	O
sensitization	NN	O	O
to	NN	O	O
chemically	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
depended	NN	O	O
on	NN	O	O
high-level	NN	O	O
E1A	NN	O	B-protein
oncoprotein	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Loss	NN	O	O
of	NN	O	O
both	NN	O	O
cytolytic	NN	O	O
susceptibility	NN	O	O
and	NN	O	O
sensitization	NN	O	O
to	NN	O	O
chemically	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
coselected	NN	O	O
during	NN	O	O
in	NN	O	O
vivo	NN	O	O
selection	NN	O	O
of	NN	O	O
E1A-positive	NN	O	B-cell_type
sarcoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
increased	NN	O	O
tumorigenicity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
E1A	NN	O	B-protein
mutant	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
can	NN	O	O
not	NN	O	O
bind	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
transcriptional	NN	O	I-protein
coactivator	NN	O	I-protein
,	NN	O	O
p300	NN	O	B-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
fail	NN	O	O
to	NN	O	O
induce	NN	O	O
cytolytic	NN	O	O
susceptibility	NN	O	O
also	NN	O	O
failed	NN	O	O
to	NN	O	O
sensitize	NN	O	O
cells	NN	O	O
to	NN	O	O
injury-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
E1A	NN	O	B-protein
induces	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
killer	NN	O	O
cell-induced	NN	O	O
lysis	NN	O	O
through	NN	O	O
sensitization	NN	O	O
of	NN	O	O
cells	NN	O	O
to	NN	O	O
injury-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

TCL1	NN	O	B-DNA
oncogene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
AIDS-related	NN	O	O
lymphomas	NN	O	O
and	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
.	NN	O	O

AIDS-related	NN	O	O
non-Hodgkin	NN	O	O
's	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
AIDS	NN	O	O
NHL	NN	O	O
)	NN	O	O
comprises	NN	O	O
a	NN	O	O
diverse	NN	O	O
and	NN	O	O
heterogeneous	NN	O	O
group	NN	O	O
of	NN	O	O
high-grade	NN	O	O
B	NN	O	O
cell	NN	O	O
tumors	NN	O	O
.	NN	O	O

Certain	NN	O	O
classes	NN	O	O
of	NN	O	O
AIDS	NN	O	O
NHL	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
alterations	NN	O	O
in	NN	O	O
oncogenes	NN	O	B-DNA
or	NN	O	O
tumor-suppressor	NN	O	B-DNA
genes	NN	O	I-DNA
or	NN	O	O
infections	NN	O	O
by	NN	O	O
oncogenic	NN	O	O
herpesviruses	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
clinically	NN	O	O
significant	NN	O	O
class	NN	O	O
of	NN	O	O
AIDS	NN	O	O
NHL	NN	O	O
designated	NN	O	O
immunoblastic	NN	O	O
lymphoma	NN	O	O
plasmacytoid	NN	O	O
(	NN	O	O
AIDS	NN	O	O
IBLP	NN	O	O
)	NN	O	O
lacks	NN	O	O
any	NN	O	O
consistent	NN	O	O
genetic	NN	O	O
alterations	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
the	NN	O	O
TCL1	NN	O	B-DNA
oncogene	NN	O	I-DNA
from	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
AIDS	NN	O	B-DNA
IBLP-associated	NN	O	I-DNA
cDNA	NN	O	I-DNA
fragments	NN	O	I-DNA
generated	NN	O	O
by	NN	O	O
subtractive	NN	O	O
hybridization	NN	O	O
with	NN	O	O
non-AIDS	NN	O	O
IBLP	NN	O	O
.	NN	O	O

Aberrant	NN	O	O
TCL1	NN	O	B-protein
expression	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia/lymphoma	NN	O	O
development	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
expression	NN	O	O
also	NN	O	O
has	NN	O	O
been	NN	O	O
seen	NN	O	O
in	NN	O	O
many	NN	O	O
established	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
tumor	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
TCL1	NN	O	B-protein
expression	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
AIDS	NN	O	O
NHL	NN	O	O
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
TCL1	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
AIDS	NN	O	O
IBLP	NN	O	O
tumors	NN	O	O
examined	NN	O	O
.	NN	O	O

TCL1	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
tumor	NN	O	O
cells	NN	O	O
in	NN	O	O
AIDS	NN	O	O
IBLP	NN	O	O
tissue	NN	O	O
samples	NN	O	O
analyzed	NN	O	O
with	NN	O	O
immunohistochemical	NN	O	O
staining	NN	O	O
.	NN	O	O

Hyperplastic	NN	O	O
lymph	NN	O	O
node	NN	O	O
and	NN	O	O
tonsil	NN	O	O
also	NN	O	O
exhibit	NN	O	O
strong	NN	O	O
TCL1	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
in	NN	O	O
mantle	NN	O	B-cell_type
zone	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
rare	NN	O	O
interfollicular	NN	O	B-cell_type
zone	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
follicle-center	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
centroblasts	NN	O	B-cell_type
and	NN	O	O
centrocytes	NN	O	B-cell_type
)	NN	O	O
show	NN	O	O
weaker	NN	O	O
expression	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
establish	NN	O	O
TCL1	NN	O	B-protein
as	NN	O	O
the	NN	O	O
most	NN	O	O
prevalent	NN	O	O
of	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
surveyed	NN	O	O
oncogenes	NN	O	B-DNA
associated	NN	O	O
with	NN	O	O
AIDS	NN	O	O
IBLP	NN	O	O
.	NN	O	O

They	NN	O	O
also	NN	O	O
indicate	NN	O	O
that	NN	O	O
abundant	NN	O	O
TCL1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
quiescent	NN	O	B-cell_type
mantle	NN	O	I-cell_type
zone	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
down-regulated	NN	O	O
in	NN	O	O
activated	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
follicular	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
parallel	NN	O	O
to	NN	O	O
the	NN	O	O
known	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
BCL-2	NN	O	B-protein
.	NN	O	O

High-level	NN	O	O
expression	NN	O	O
in	NN	O	O
nonproliferating	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
suggests	NN	O	O
that	NN	O	O
TCL1	NN	O	B-protein
may	NN	O	O
function	NN	O	O
in	NN	O	O
protecting	NN	O	O
naive	NN	O	B-cell_type
preactivated	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

IL-2	NN	O	B-protein
-independent	NN	O	O
activation	NN	O	O
and	NN	O	O
proliferation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
induced	NN	O	O
by	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
in	NN	O	O
T	NN	O	O
cell	NN	O	O
costimulation	NN	O	O
is	NN	O	O
firmly	NN	O	O
established	NN	O	O
,	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
it	NN	O	O
exerts	NN	O	O
its	NN	O	O
costimulatory	NN	O	O
actions	NN	O	O
are	NN	O	O
less	NN	O	O
clear	NN	O	O
.	NN	O	O

In	NN	O	O
many	NN	O	O
circumstances	NN	O	O
it	NN	O	O
is	NN	O	O
difficult	NN	O	O
to	NN	O	O
distinguish	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
from	NN	O	O
subsequent	NN	O	O
actions	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
on	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
a	NN	O	O
model	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
costimulation	NN	O	O
using	NN	O	O
PMA	NN	O	O
plus	NN	O	O
the	NN	O	O
natural	NN	O	B-protein
ligand	NN	O	I-protein
CD80	NN	O	I-protein
that	NN	O	O
resulted	NN	O	O
in	NN	O	O
very	NN	O	O
limited	NN	O	O
stimulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
both	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
and	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
stimulation	NN	O	O
.	NN	O	O

Promoter	NN	O	O
assays	NN	O	O
revealed	NN	O	O
CD28	NN	O	B-protein
-dependent	NN	O	O
effects	NN	O	O
on	NN	O	O
both	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
on	NN	O	O
NF-AT	NN	O	B-protein
or	NN	O	O
the	NN	O	O
intact	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
was	NN	O	O
completely	NN	O	O
resistant	NN	O	O
to	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
the	NN	O	O
immunosuppressant	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
.	NN	O	O

Moreover	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
blocking	NN	O	O
Abs	NN	O	B-protein
to	NN	O	O
both	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
this	NN	O	O
form	NN	O	O
of	NN	O	O
costimulation	NN	O	O
by	NN	O	O
CD28	NN	O	B-protein
was	NN	O	O
independent	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
stimulating	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
blasts	NN	O	I-cell_type
with	NN	O	O
CD80	NN	O	B-protein
alone	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
limited	NN	O	O
requirement	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
this	NN	O	O
system	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
CD28	NN	O	B-protein
costimulation	NN	O	O
can	NN	O	O
cause	NN	O	O
substantial	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
driven	NN	O	O
by	NN	O	O
a	NN	O	O
soluble	NN	O	B-protein
factor	NN	O	I-protein
independent	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
transactivation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Epstein-Barr	NN	O	B-DNA
virus	NN	O	I-DNA
latency	NN	O	I-DNA
BamHI-Q	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
positively	NN	O	O
regulated	NN	O	O
by	NN	O	O
STATs	NN	O	B-protein
and	NN	O	O
Zta	NN	O	B-protein
interference	NN	O	O
with	NN	O	O
JAK	NN	O	B-protein
/STAT	NN	O	B-protein
activation	NN	O	O
leads	NN	O	O
to	NN	O	O
loss	NN	O	O
of	NN	O	O
BamHI-Q	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

In	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
-associated	NN	O	O
tumors	NN	O	O
in	NN	O	O
nonimmunocompromised	NN	O	O
patients	NN	O	O
,	NN	O	O
EBV	NN	O	O
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
highly	NN	O	O
restricted	NN	O	O
.	NN	O	O

EBV-encoded	NN	O	B-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
(	NN	O	I-protein
EBNA	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
immunogenic	NN	O	O
and	NN	O	O
proliferative	NN	O	O
EBNAs	NN	O	B-protein
are	NN	O	O
not	NN	O	O
.	NN	O	O

This	NN	O	O
pattern	NN	O	O
of	NN	O	O
EBNA	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
generated	NN	O	O
by	NN	O	O
usage	NN	O	O
of	NN	O	O
the	NN	O	O
BamHI-Q	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
Qp	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
determined	NN	O	O
that	NN	O	O
the	NN	O	O
JAK	NN	O	B-protein
/STAT	NN	O	B-protein
pathway	NN	O	O
positively	NN	O	O
regulates	NN	O	O
Qp	NN	O	B-DNA
activity	NN	O	O
.	NN	O	O

In	NN	O	O
transient-transfection	NN	O	O
assays	NN	O	O
,	NN	O	O
a	NN	O	O
Qp-CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
was	NN	O	O
activated	NN	O	O
by	NN	O	O
cotransfected	NN	O	O
JAK-1	NN	O	B-protein
and	NN	O	O
by	NN	O	O
treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
IL-6	NN	O	B-protein
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
Qp	NN	O	B-DNA
to	NN	O	O
bind	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
was	NN	O	O
directly	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility-shift	NN	O	O
assay	NN	O	O
,	NN	O	O
and	NN	O	O
mutation	NN	O	O
of	NN	O	O
potential	NN	O	O
STAT-binding	NN	O	B-DNA
sites	NN	O	I-DNA
reduced	NN	O	O
Qp	NN	O	B-DNA
responsiveness	NN	O	O
to	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
JAK	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
STATs	NN	O	B-protein
in	NN	O	O
Qp	NN	O	B-DNA
function	NN	O	O
,	NN	O	O
Qp	NN	O	B-DNA
using	NN	O	O
Burkitt	NN	O	B-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
Rael	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
cultured	NN	O	B-cell_type
nasopharyngeal	NN	O	I-cell_type
carcinoma	NN	O	I-cell_type
(	NN	O	I-cell_type
NPC	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
contained	NN	O	O
nuclear	NN	O	O
STAT	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
whether	NN	O	O
the	NN	O	O
inability	NN	O	O
to	NN	O	O
maintain	NN	O	O
EBV-positive	NN	O	B-cell_line
NPC	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
in	NN	O	O
culture	NN	O	O
was	NN	O	O
related	NN	O	O
to	NN	O	O
Qp	NN	O	B-DNA
activity	NN	O	O
.	NN	O	O

Passaging	NN	O	O
of	NN	O	O
the	NN	O	O
NPC	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
HK666	NN	O	I-cell_line
led	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
BZLF1	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
Zta	NN	O	B-protein
and	NN	O	O
loss	NN	O	O
of	NN	O	O
Qp	NN	O	B-DNA
function	NN	O	O
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
assays	NN	O	O
linked	NN	O	O
Zta	NN	O	B-protein
expression	NN	O	O
to	NN	O	O
the	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
Qp	NN	O	B-DNA
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
Zta	NN	O	B-protein
reduced	NN	O	O
Qp	NN	O	B-DNA
activity	NN	O	O
in	NN	O	O
reporter	NN	O	O
assays	NN	O	O
.	NN	O	O

This	NN	O	O
negative	NN	O	O
regulation	NN	O	O
required	NN	O	O
Zta	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
Zta	NN	O	B-protein
up-regulation	NN	O	O
of	NN	O	O
p53	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
p53	NN	O	B-protein
-mediated	NN	O	O
interference	NN	O	O
with	NN	O	O
JAK	NN	O	B-protein
/STAT	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
Qp	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
data	NN	O	O
imply	NN	O	O
that	NN	O	O
JAK	NN	O	B-protein
/STAT	NN	O	B-protein
signaling	NN	O	O
has	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
EBV-associated	NN	O	O
malignancies	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reactive	NN	O	O
oxygen	NN	O	O
intermediate-release	NN	O	O
of	NN	O	O
fibre-exposed	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
increases	NN	O	O
inflammatory	NN	O	O
cytokine-mRNA	NN	O	B-RNA
level	NN	O	O
,	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
co-cultured	NN	O	B-cell_line
bronchial	NN	O	I-cell_line
epithelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
BEAS-2B	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

Some	NN	O	O
pulmonary	NN	O	O
diseases	NN	O	O
like	NN	O	O
bronchitis	NN	O	O
or	NN	O	O
asthma	NN	O	O
bronchiale	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
inflammatory	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
bronchial	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
are	NN	O	O
located	NN	O	O
directly	NN	O	O
in	NN	O	O
the	NN	O	O
surrounding	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
so	NN	O	O
that	NN	O	O
we	NN	O	O
suppose	NN	O	O
an	NN	O	O
interaction	NN	O	O
between	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
regarding	NN	O	O
to	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
.	NN	O	O

For	NN	O	O
measuring	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
to	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
by	NN	O	O
bronchial	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
established	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
model	NN	O	O
of	NN	O	O
co-cultured	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
(	NN	O	O
BM	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
BEAS-2B	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
a	NN	O	O
transwell	NN	O	O
system	NN	O	O
(	NN	O	O
Costar	NN	O	O
)	NN	O	O
.	NN	O	O

BM	NN	O	B-cell_type
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
Chrysotile	NN	O	O
B	NN	O	O
and	NN	O	O
soot	NN	O	O
particle	NN	O	O
FR	NN	O	O
101	NN	O	O
in	NN	O	O
a	NN	O	O
concentration	NN	O	O
of	NN	O	O
100	NN	O	O
microg/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
.	NN	O	O

After	NN	O	O
up	NN	O	O
to	NN	O	O
90	NN	O	O
min	NN	O	O
exposure	NN	O	O
time	NN	O	O
ELISA	NN	O	O
,	NN	O	O
EMSA	NN	O	O
(	NN	O	O
electromobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
)	NN	O	O
and	NN	O	O
RT-PCR	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
measure	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
protein	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
cytokine	NN	O	B-protein
(	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
specific	NN	O	O
mRNA	NN	O	B-RNA
levels	NN	O	O
in	NN	O	O
BEAS-2B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
observed	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
activity	NN	O	O
(	NN	O	O
up	NN	O	O
to	NN	O	O
1.8	NN	O	O
+/-	NN	O	O
0.5-fold	NN	O	O
)	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
protein	NN	O	O
activity	NN	O	O
in	NN	O	O
BEAS-2B	NN	O	B-cell_type
cells	NN	O	I-cell_type
after	NN	O	O
particle	NN	O	O
or	NN	O	O
fibre	NN	O	O
exposure	NN	O	O
of	NN	O	O
co-cultured	NN	O	B-cell_line
BM	NN	O	I-cell_line
.	NN	O	O

Consecutive	NN	O	O
IL-1beta-	NN	O	B-RNA
,	NN	O	I-RNA
IL-6-	NN	O	I-RNA
and	NN	O	I-RNA
TNF-alpha-mRNA	NN	O	I-RNA
were	NN	O	O
elevated	NN	O	O
(	NN	O	O
up	NN	O	O
to	NN	O	O
1.9	NN	O	O
+/-	NN	O	O
0.58-fold	NN	O	O
)	NN	O	O
.	NN	O	O

Protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
cytokine-specific	NN	O	B-RNA
mRNA	NN	O	I-RNA
were	NN	O	O
inhibited	NN	O	O
by	NN	O	O
antioxidants	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
a	NN	O	O
ROI-dependent	NN	O	O
NF-kappaB	NN	O	B-protein
mediated	NN	O	O
transcription	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
in	NN	O	O
bronchial	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
by	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
NFAT-1	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
HIV-1	NN	O	O
infected	NN	O	O
individuals	NN	O	O
produce	NN	O	O
a	NN	O	O
soluble	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
efficiently	NN	O	O
suppresses	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
long	NN	O	O
terminal	NN	O	O
repeat	NN	O	O
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
to	NN	O	O
mitogenic	NN	O	O
or	NN	O	O
Tat	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
is	NN	O	O
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
suppressive	NN	O	O
action	NN	O	O
of	NN	O	O
a	NN	O	O
Herpesvirus	NN	O	B-cell_line
saimiri	NN	O	I-cell_line
(	NN	O	I-cell_line
HVS	NN	O	I-cell_line
)	NN	O	I-cell_line
-transformed	NN	O	I-cell_line
CD8+	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
from	NN	O	O
an	NN	O	O
HIV-infected	NN	O	O
individual	NN	O	O
and	NN	O	O
supernatants	NN	O	O
from	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
HIV-1-infected	NN	O	O
asymptomatic	NN	O	O
subjects	NN	O	O
(	NN	O	O
CD4+	NN	O	B-protein
>	NN	O	O
350/microliters	NN	O	O
)	NN	O	O
.	NN	O	O

Mutagenesis	NN	O	O
of	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
or	NN	O	I-DNA
Sp-1	NN	O	I-DNA
elements	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
resulted	NN	O	O
in	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
supernatants	NN	O	O
to	NN	O	O
inhibit	NN	O	O
Tat-	NN	O	O
or	NN	O	O
mitogen-mediated	NN	O	O
LTR	NN	O	O
transcription	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
HIV-1	NN	O	B-protein
Tat	NN	O	I-protein
by	NN	O	O
a	NN	O	O
LTR	NN	O	B-DNA
in	NN	O	O
which	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
(	NN	O	I-DNA
IL	NN	O	I-DNA
)	NN	O	I-DNA
-2	NN	O	I-DNA
homology	NN	O	I-DNA
NFAT-1	NN	O	I-DNA
region	NN	O	I-DNA
was	NN	O	O
mutated	NN	O	O
resulted	NN	O	O
in	NN	O	O
almost	NN	O	O
complete	NN	O	O
elimination	NN	O	O
of	NN	O	O
suppression	NN	O	O
by	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
when	NN	O	O
the	NN	O	O
NFAT-1	NN	O	B-DNA
mutant	NN	O	I-DNA
LTR	NN	O	I-DNA
was	NN	O	O
activated	NN	O	O
by	NN	O	O
mitogen	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
gene	NN	O	O
expression	NN	O	O
directed	NN	O	O
by	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
elements	NN	O	I-DNA
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
CD8	NN	O	B-protein
+	NN	O	O
cell-derived	NN	O	O
supernatants	NN	O	O
(	NN	O	O
Copeland	NN	O	O
et	NN	O	O
al.	NN	O	O
,	NN	O	O
AIDS	NN	O	O
Res	NN	O	O
Hum	NN	O	O
Retroviruses	NN	O	O
,	NN	O	O
1995	NN	O	O
;	NN	O	O
11	NN	O	O
:	NN	O	O
1321-1326	NN	O	O
)	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
mitogenic	NN	O	O
activation	NN	O	O
,	NN	O	O
mediated	NN	O	O
primarily	NN	O	O
through	NN	O	O
the	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
susceptible	NN	O	O
to	NN	O	O
CD8	NN	O	B-protein
-mediated	NN	O	O
inhibition	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Tat	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
may	NN	O	O
rely	NN	O	O
upon	NN	O	O
a	NN	O	O
different	NN	O	O
pathway	NN	O	O
that	NN	O	O
is	NN	O	O
NFAT-1	NN	O	B-protein
dependent	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
by	NN	O	O
anetholdithiolthione	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
redox	NN	O	B-protein
sensitive	NN	O	I-protein
cytosolic	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Redox	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
immuno-deficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
effective	NN	O	O
strategy	NN	O	O
for	NN	O	O
acquired	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
therapy	NN	O	O
.	NN	O	O

Anetholdithiolthione	NN	O	O
(	NN	O	O
ADT	NN	O	O
,	NN	O	O
5-	NN	O	O
[	NN	O	O
p-methoxyphenyl	NN	O	O
]	NN	O	O
-3H-1	NN	O	O
,	NN	O	O
2-dithiol-3-thione	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
antioxidant	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
protect	NN	O	O
against	NN	O	O
acetaminophen-	NN	O	O
and	NN	O	O
CCl4-induced	NN	O	O
hepatotoxicity	NN	O	O
,	NN	O	O
lipid	NN	O	O
peroxidation	NN	O	O
,	NN	O	O
radiation	NN	O	O
injury	NN	O	O
,	NN	O	O
and	NN	O	O
also	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
clinically	NN	O	O
as	NN	O	O
an	NN	O	O
anti-choleretic	NN	O	O
agent	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
ADT	NN	O	O
pretreatment	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
stimuli	NN	O	O
such	NN	O	O
as	NN	O	O
H2O2	NN	O	O
,	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
or	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
.	NN	O	O

PMA	NN	O	O
and	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
induced	NN	O	O
activation	NN	O	O
of	NN	O	O
(	NN	O	B-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
human	NN	O	O
Jurkat	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
was	NN	O	O
partially	NN	O	O
inhibited	NN	O	O
by	NN	O	O
ADT	NN	O	O
(	NN	O	O
0.1	NN	O	O
mM	NN	O	O
)	NN	O	O
pretreatment	NN	O	O
.	NN	O	O

ADT	NN	O	O
(	NN	O	O
0.1	NN	O	O
mM	NN	O	O
)	NN	O	O
also	NN	O	O
inhibited	NN	O	O
H2O2	NN	O	O
induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
peroxide	NN	O	B-cell_line
sensitive	NN	O	I-cell_line
human	NN	O	I-cell_line
Wurzburg	NN	O	I-cell_line
T-cells	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
ADT	NN	O	O
treated	NN	O	O
Wurzburg	NN	O	B-cell_line
cells	NN	O	I-cell_line
had	NN	O	O
significantly	NN	O	O
higher	NN	O	O
glutathione	NN	O	O
levels	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
untreated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

H2O2	NN	O	O
induced	NN	O	O
lipid	NN	O	O
peroxidation	NN	O	O
in	NN	O	O
Wurzburg	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
remarkably	NN	O	O
inhibited	NN	O	O
by	NN	O	O
ADT	NN	O	O
pretreatment	NN	O	O
.	NN	O	O

ADT	NN	O	O
,	NN	O	O
a	NN	O	O
pro-glutathione	NN	O	O
antioxidant	NN	O	O
,	NN	O	O
was	NN	O	O
observed	NN	O	O
to	NN	O	O
be	NN	O	O
capable	NN	O	O
of	NN	O	O
modulating	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
endothelial	NN	O	B-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
induction	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
endothelial	NN	O	B-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
by	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
can	NN	O	O
occur	NN	O	O
independently	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
and	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
PTK	NN	O	B-protein
)	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
by	NN	O	O
interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
on	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
PTK	NN	O	B-protein
inhibitors	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
or	NN	O	O
genistein	NN	O	O
suppress	NN	O	O
induction	NN	O	O
of	NN	O	O
endothelial	NN	O	O
VCAM-1	NN	O	B-protein
and	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
subsequent	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
TNF	NN	O	B-protein
.	NN	O	O

Inhibition	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
specific	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
following	NN	O	O
PTK	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
mobilization	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
VCAM-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
pathophysiological	NN	O	O
conditions	NN	O	O
that	NN	O	O
involve	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
these	NN	O	O
molecules	NN	O	O
(	NN	O	O
e.g.	NN	O	O
inflammation	NN	O	O
and	NN	O	O
atherosclerosis	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
C-zeta	NN	O	I-protein
mediates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
immunodeficiency	NN	O	I-cell_type
virus-infected	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
regulating	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
persistence	NN	O	O
in	NN	O	O
a	NN	O	O
major	NN	O	O
cell	NN	O	O
reservoir	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
macrophage	NN	O	B-cell_type
remain	NN	O	O
unknown	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
selectively	NN	O	O
activated	NN	O	O
following	NN	O	O
HIV	NN	O	O
infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
little	NN	O	O
information	NN	O	O
as	NN	O	O
to	NN	O	O
what	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
mediate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
monocytes-macrophages	NN	O	B-cell_type
is	NN	O	O
available	NN	O	O
,	NN	O	O
our	NN	O	O
previous	NN	O	O
work	NN	O	O
indicated	NN	O	O
that	NN	O	O
classical	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	I-protein
PKC	NN	O	I-protein
)	NN	O	I-protein
isoenzymes	NN	O	I-protein
were	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
HIV-mediated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
focused	NN	O	O
on	NN	O	O
atypical	NN	O	O
PKC	NN	O	B-protein
isoenzymes	NN	O	I-protein
.	NN	O	O

PKC-zeta	NN	O	B-protein
belongs	NN	O	O
to	NN	O	O
this	NN	O	O
family	NN	O	O
and	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
step	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
other	NN	O	O
cell	NN	O	O
systems	NN	O	O
.	NN	O	O

Immunoblotting	NN	O	O
experiments	NN	O	O
with	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
demonstrate	NN	O	O
that	NN	O	O
PKC-zeta	NN	O	B-protein
is	NN	O	O
present	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
expression	NN	O	O
can	NN	O	O
be	NN	O	O
downmodulated	NN	O	O
by	NN	O	O
antisense	NN	O	O
oligonucleotides	NN	O	O
(	NN	O	O
AO	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
HIV-mediated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
is	NN	O	O
selectively	NN	O	O
reduced	NN	O	O
by	NN	O	O
AO	NN	O	O
to	NN	O	O
PKC-zeta	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
cotransfection	NN	O	O
of	NN	O	O
a	NN	O	O
negative	NN	O	O
dominant	NN	O	O
molecule	NN	O	O
of	NN	O	O
PKC-zeta	NN	O	B-protein
(	NN	O	O
PKC-zeta	NN	O	B-protein
mut	NN	O	I-protein
)	NN	O	O
with	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
reporter	NN	O	O
genes	NN	O	O
selectively	NN	O	O
inhibits	NN	O	O
the	NN	O	O
HIV	NN	O	O
-but	NN	O	O
not	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate-	NN	O	O
or	NN	O	O
lipopolysaccharide-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

That	NN	O	O
PKC-zeta	NN	O	B-protein
is	NN	O	O
specific	NN	O	O
in	NN	O	O
regulating	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
concluded	NN	O	O
from	NN	O	O
the	NN	O	O
inability	NN	O	O
of	NN	O	O
PKC-zeta	NN	O	B-protein
(	NN	O	I-protein
mut	NN	O	I-protein
)	NN	O	I-protein
to	NN	O	O
interfere	NN	O	O
with	NN	O	O
the	NN	O	O
basal	NN	O	O
or	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate-inducible	NN	O	O
CREB	NN	O	B-protein
-or	NN	O	O
AP1	NN	O	B-protein
-dependent	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

Lastly	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
selective	NN	O	O
inhibition	NN	O	O
of	NN	O	O
p24	NN	O	B-protein
production	NN	O	O
by	NN	O	O
HIV-infected	NN	O	B-cell_type
human	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
when	NN	O	O
treated	NN	O	O
with	NN	O	O
AO	NN	O	O
to	NN	O	O
PKC-zeta	NN	O	B-protein
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
atypical	NN	O	B-protein
PKC	NN	O	I-protein
isoenzymes	NN	O	I-protein
,	NN	O	O
including	NN	O	O
PKC-zeta	NN	O	B-protein
,	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
by	NN	O	O
which	NN	O	O
HIV	NN	O	O
infection	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
Bcl-x	NN	O	B-protein
(	NN	O	I-protein
L	NN	O	I-protein
)	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
Tax	NN	O	I-protein
through	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
apoptosis-resistant	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
transfectants	NN	O	I-cell_line
with	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
Tax	NN	O	B-protein
is	NN	O	O
thought	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
pivotal	NN	O	O
role	NN	O	O
in	NN	O	O
immortalization	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
protected	NN	O	O
the	NN	O	O
mouse	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
CTLL-2	NN	O	I-cell_line
against	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
deprivation	NN	O	O
and	NN	O	O
converted	NN	O	O
its	NN	O	O
growth	NN	O	O
from	NN	O	O
being	NN	O	O
IL-2	NN	O	B-protein
dependent	NN	O	O
to	NN	O	O
being	NN	O	O
IL-2	NN	O	B-protein
independent	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
bcl-xl	NN	O	B-protein
but	NN	O	O
not	NN	O	O
bcl-2	NN	O	B-protein
,	NN	O	O
bcl-xs	NN	O	B-protein
,	NN	O	O
bak	NN	O	B-protein
,	NN	O	O
bad	NN	O	B-protein
,	NN	O	O
or	NN	O	O
bax	NN	O	B-protein
was	NN	O	O
associated	NN	O	O
with	NN	O	O
apoptosis	NN	O	O
resistance	NN	O	O
after	NN	O	O
IL-2	NN	O	B-protein
deprivation	NN	O	O
in	NN	O	O
CTLL-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
expressed	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

Transient-transfection	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
bcl-x	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
transactivated	NN	O	O
by	NN	O	O
wild-type	NN	O	B-protein
Tax	NN	O	I-protein
.	NN	O	O

Similar	NN	O	O
effects	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
mutant	NN	O	O
Tax	NN	O	B-protein
retaining	NN	O	O
transactivating	NN	O	O
ability	NN	O	O
through	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Deletion	NN	O	O
or	NN	O	O
substitution	NN	O	O
of	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
NF-kappaB	NN	O	I-protein
binding	NN	O	I-protein
site	NN	O	I-protein
identified	NN	O	O
in	NN	O	O
the	NN	O	O
bcl-x	NN	O	B-DNA
promoter	NN	O	I-DNA
significantly	NN	O	O
decreased	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
transactivation	NN	O	O
.	NN	O	O

This	NN	O	O
NF-kappaB-like	NN	O	B-DNA
element	NN	O	I-DNA
was	NN	O	O
able	NN	O	O
to	NN	O	O
form	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
NF-kappaB	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
bcl-x	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
also	NN	O	O
diminished	NN	O	O
by	NN	O	O
the	NN	O	O
mutant	NN	O	B-protein
IkappaBalpha	NN	O	I-protein
,	NN	O	O
which	NN	O	O
specifically	NN	O	O
inhibits	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
Bcl-x	NN	O	B-protein
(	NN	O	I-protein
L	NN	O	I-protein
)	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
through	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
pathway	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
CTLL-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
after	NN	O	O
IL-2	NN	O	B-protein
deprivation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
beta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
important	NN	O	O
for	NN	O	O
recruitment	NN	O	O
of	NN	O	O
erythroid	NN	O	B-protein
Kruppel-like	NN	O	I-protein
factor	NN	O	I-protein
to	NN	O	O
the	NN	O	O
locus	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Erythroid	NN	O	B-protein
Kruppel-like	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EKLF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
CACCC	NN	O	O
box	NN	O	O
in	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
adult	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
was	NN	O	O
recently	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
EKLF	NN	O	B-protein
is	NN	O	O
also	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
locus	NN	O	I-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
LCR	NN	O	B-DNA
)	NN	O	O
5'HS3	NN	O	B-DNA
.	NN	O	O

Some	NN	O	O
evidence	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
LCR	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
interact	NN	O	O
in	NN	O	O
adult	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
network	NN	O	O
of	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
that	NN	O	O
exists	NN	O	O
between	NN	O	O
these	NN	O	O
two	NN	O	O
elements	NN	O	O
may	NN	O	O
regulate	NN	O	O
how	NN	O	O
EKLF	NN	O	B-protein
is	NN	O	O
recruited	NN	O	O
to	NN	O	O
the	NN	O	O
LCR	NN	O	B-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
use	NN	O	O
the	NN	O	O
PIN*POINT	NN	O	O
assay	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
on	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
EKLF	NN	O	B-protein
to	NN	O	O
5'HS2	NN	O	B-DNA
and	NN	O	O
5'HS3	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
LCR	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
recruitment	NN	O	O
of	NN	O	O
EKLF	NN	O	B-protein
to	NN	O	O
5'HS2	NN	O	B-DNA
requires	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
recruitment	NN	O	O
to	NN	O	O
5'HS3	NN	O	B-DNA
depends	NN	O	O
on	NN	O	O
the	NN	O	O
CACCC	NN	O	O
and	NN	O	O
TATA	NN	O	B-DNA
boxes	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
recruitment	NN	O	O
of	NN	O	O
EKLF	NN	O	B-protein
to	NN	O	O
5'HS3	NN	O	B-DNA
only	NN	O	O
occurred	NN	O	O
in	NN	O	O
beta-globin-expressing	NN	O	B-cell_type
murine	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
recruitment	NN	O	O
of	NN	O	O
EKLF	NN	O	B-protein
to	NN	O	O
5'HS2	NN	O	B-DNA
occurred	NN	O	O
in	NN	O	O
both	NN	O	O
gamma-globin-expressing	NN	O	B-cell_line
K562	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
murine	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Unlike	NN	O	O
EKLF	NN	O	B-protein
,	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
also	NN	O	O
binds	NN	O	O
to	NN	O	O
CACCC	NN	O	O
boxes	NN	O	O
,	NN	O	O
is	NN	O	O
not	NN	O	O
recruited	NN	O	O
to	NN	O	O
5'HS3	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
also	NN	O	O
examined	NN	O	O
how	NN	O	O
one	NN	O	O
5'HS	NN	O	B-DNA
affects	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
EKLF	NN	O	B-protein
to	NN	O	O
another	NN	O	O
5'HS	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
EKLF	NN	O	B-protein
to	NN	O	O
5'HS3	NN	O	B-DNA
depends	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
5'HS2	NN	O	B-DNA
in	NN	O	O
cis	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
recruitment	NN	O	O
to	NN	O	O
5'HS2	NN	O	B-DNA
does	NN	O	O
not	NN	O	O
depend	NN	O	O
on	NN	O	O
5'HS3	NN	O	B-DNA
.	NN	O	O

Based	NN	O	O
on	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
we	NN	O	O
present	NN	O	O
a	NN	O	O
model	NN	O	O
that	NN	O	O
illustrates	NN	O	O
how	NN	O	O
EKLF	NN	O	B-protein
may	NN	O	O
be	NN	O	O
recruited	NN	O	O
to	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
locus	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Estrogen	NN	O	O
decreases	NN	O	O
TNF	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
blocking	NN	O	O
JNK	NN	O	B-protein
activity	NN	O	O
and	NN	O	O
the	NN	O	O
resulting	NN	O	O
production	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
JunD	NN	O	B-protein
.	NN	O	O

Central	NN	O	O
to	NN	O	O
the	NN	O	O
bone-sparing	NN	O	O
effect	NN	O	O
of	NN	O	O
estrogen	NN	O	O
(	NN	O	O
E	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
)	NN	O	O
is	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
block	NN	O	O
the	NN	O	O
monocytic	NN	O	O
production	NN	O	O
of	NN	O	O
the	NN	O	O
osteoclastogenic	NN	O	B-protein
cytokine	NN	O	I-protein
TNF-alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
E	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
downregulates	NN	O	O
TNF	NN	O	B-protein
production	NN	O	O
is	NN	O	O
presently	NN	O	O
unknown	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
studies	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
an	NN	O	O
E	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
receptor-negative	NN	O	O
line	NN	O	O
,	NN	O	O
suggest	NN	O	O
that	NN	O	O
E	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
inhibits	NN	O	O
TNF	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
through	NN	O	O
an	NN	O	O
effect	NN	O	O
mediated	NN	O	O
by	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
beta	NN	O	I-protein
(	NN	O	O
ERbeta	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
report	NN	O	O
that	NN	O	O
in	NN	O	O
RAW	NN	O	B-cell_line
264.7	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
an	NN	O	O
E	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
receptor-positive	NN	O	O
murine	NN	O	O
monocytic	NN	O	O
line	NN	O	O
,	NN	O	O
E	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
downregulates	NN	O	O
cytokine-induced	NN	O	O
TNF	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
decreasing	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
Jun	NN	O	B-protein
NH	NN	O	I-protein
(	NN	O	I-protein
2	NN	O	I-protein
)	NN	O	I-protein
-terminal	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
resulting	NN	O	O
diminished	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
JunD	NN	O	B-protein
at	NN	O	O
their	NN	O	O
NH	NN	O	B-protein
(	NN	O	I-protein
2	NN	O	I-protein
)	NN	O	I-protein
-termini	NN	O	I-protein
decreases	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
these	NN	O	O
nuclear	NN	O	O
proteins	NN	O	O
to	NN	O	O
autostimulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
c-Jun	NN	O	B-DNA
and	NN	O	I-DNA
JunD	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
thus	NN	O	O
leading	NN	O	O
to	NN	O	O
lower	NN	O	O
production	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
JunD	NN	O	B-protein
.	NN	O	O

The	NN	O	O
consequent	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
levels	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
JunD	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
diminished	NN	O	O
binding	NN	O	O
of	NN	O	O
c-Jun/c-Fos	NN	O	B-protein
and	NN	O	O
JunD/c-Fos	NN	O	B-protein
heterodimers	NN	O	I-protein
to	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
,	NN	O	O
thus	NN	O	O
,	NN	O	O
to	NN	O	O
decreased	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Cutting	NN	O	O
edge	NN	O	O
:	NN	O	O
TCR	NN	O	B-protein
stimulation	NN	O	O
by	NN	O	O
antibody	NN	O	O
and	NN	O	O
bacterial	NN	O	O
superantigen	NN	O	O
induces	NN	O	O
Stat3	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Recent	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
TCR/CD3	NN	O	B-protein
stimulation	NN	O	O
induces	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat5	NN	O	B-protein
in	NN	O	O
murine	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
CD3	NN	O	O
ligation	NN	O	O
by	NN	O	O
mAb	NN	O	O
and	NN	O	O
Staphylococcal	NN	O	O
enterotoxin	NN	O	O
(	NN	O	O
SE	NN	O	O
)	NN	O	O
induce	NN	O	O
a	NN	O	O
rapid	NN	O	O
,	NN	O	O
gradually	NN	O	O
accumulating	NN	O	O
,	NN	O	O
long-lasting	NN	O	O
tyrosine	NN	O	O
,	NN	O	O
and	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
(	NN	O	O
but	NN	O	O
not	NN	O	O
Stat5	NN	O	B-protein
)	NN	O	O
in	NN	O	O
allogen-specific	NN	O	B-cell_line
human	NN	O	I-cell_line
CD4+	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
induces	NN	O	O
a	NN	O	O
rapid	NN	O	O
and	NN	O	O
transient	NN	O	O
tyrosine	NN	O	O
and	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
.	NN	O	O

Compared	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
CD3	NN	O	B-protein
ligation	NN	O	O
induces	NN	O	O
a	NN	O	O
delayed	NN	O	O
Stat3	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
oligonucleotide	NN	O	O
probes	NN	O	O
from	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-protein
and	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

CD3	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
is	NN	O	O
almost	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
Src	NN	O	B-protein
kinase	NN	O	I-protein
inhibitor	NN	O	I-protein
(	NN	O	O
PP1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
Stat3	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
unaffected	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
CD3	NN	O	B-protein
ligation	NN	O	O
by	NN	O	O
mAb	NN	O	B-protein
and	NN	O	O
SE	NN	O	O
triggers	NN	O	O
a	NN	O	O
rapid	NN	O	O
,	NN	O	O
PP1	NN	O	B-protein
-sensitive	NN	O	O
tyrosine	NN	O	O
and	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
TCR/CD3	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
induce	NN	O	O
Stat3	NN	O	B-protein
activation	NN	O	O
via	NN	O	O
distinct	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

Distinctive	NN	O	O
gene	NN	O	O
expression	NN	O	O
patterns	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
mammary	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
breast	NN	O	O
cancers	NN	O	O
.	NN	O	O

cDNA	NN	O	B-DNA
microarrays	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
clustering	NN	O	O
algorithm	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
patterns	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
mammary	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
growing	NN	O	O
in	NN	O	O
culture	NN	O	O
and	NN	O	O
in	NN	O	O
primary	NN	O	O
human	NN	O	O
breast	NN	O	O
tumors	NN	O	O
.	NN	O	O

Clusters	NN	O	O
of	NN	O	O
coexpressed	NN	O	B-DNA
genes	NN	O	I-DNA
identified	NN	O	O
through	NN	O	O
manipulations	NN	O	O
of	NN	O	O
mammary	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
also	NN	O	O
showed	NN	O	O
consistent	NN	O	O
patterns	NN	O	O
of	NN	O	O
variation	NN	O	O
in	NN	O	O
expression	NN	O	O
among	NN	O	O
breast	NN	O	O
tumor	NN	O	O
samples	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
immunohistochemistry	NN	O	O
with	NN	O	O
antibodies	NN	O	O
against	NN	O	O
proteins	NN	O	O
encoded	NN	O	O
by	NN	O	O
a	NN	O	O
particular	NN	O	O
gene	NN	O	O
in	NN	O	O
a	NN	O	O
cluster	NN	O	O
,	NN	O	O
the	NN	O	O
identity	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
type	NN	O	O
within	NN	O	O
the	NN	O	O
tumor	NN	O	O
specimen	NN	O	O
that	NN	O	O
contributed	NN	O	O
the	NN	O	O
observed	NN	O	O
gene	NN	O	O
expression	NN	O	O
pattern	NN	O	O
could	NN	O	O
be	NN	O	O
determined	NN	O	O
.	NN	O	O

Clusters	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
with	NN	O	O
coherent	NN	O	O
expression	NN	O	O
patterns	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
the	NN	O	O
breast	NN	O	O
tumors	NN	O	O
samples	NN	O	O
could	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
specific	NN	O	O
features	NN	O	O
of	NN	O	O
biological	NN	O	O
variation	NN	O	O
among	NN	O	O
the	NN	O	O
samples	NN	O	O
.	NN	O	O

Two	NN	O	O
such	NN	O	O
clusters	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
patterns	NN	O	O
that	NN	O	O
correlated	NN	O	O
with	NN	O	O
variation	NN	O	O
in	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
rates	NN	O	O
and	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IFN	NN	O	B-protein
-regulated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Clusters	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
expressed	NN	O	O
by	NN	O	O
stromal	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
lymphocytes	NN	O	B-cell_type
in	NN	O	O
the	NN	O	O
breast	NN	O	O
tumors	NN	O	O
also	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
this	NN	O	O
analysis	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
feasibility	NN	O	O
and	NN	O	O
usefulness	NN	O	O
of	NN	O	O
this	NN	O	O
systematic	NN	O	O
approach	NN	O	O
to	NN	O	O
studying	NN	O	O
variation	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
patterns	NN	O	O
in	NN	O	O
human	NN	O	O
cancers	NN	O	O
as	NN	O	O
a	NN	O	O
means	NN	O	O
to	NN	O	O
dissect	NN	O	O
and	NN	O	O
classify	NN	O	O
solid	NN	O	O
tumors	NN	O	O
.	NN	O	O

-DOCSTART-	O

PPARgamma	NN	O	B-protein
activation	NN	O	O
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
adipocyte	NN	O	B-DNA
fatty	NN	O	I-DNA
acid	NN	O	I-DNA
binding	NN	O	I-DNA
protein	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
peroxisome-proliferator	NN	O	B-protein
activated	NN	O	I-protein
receptor	NN	O	I-protein
gamma	NN	O	I-protein
(	NN	O	O
PPARgamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
of	NN	O	O
ligand	NN	O	B-protein
activated	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
anti-diabetic	NN	O	O
actions	NN	O	O
of	NN	O	O
the	NN	O	O
thiazolidinediones	NN	O	O
(	NN	O	O
TZDs	NN	O	O
)	NN	O	O
.	NN	O	O

PPARgamma	NN	O	B-protein
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
genes	NN	O	B-DNA
involved	NN	O	O
in	NN	O	O
lipid	NN	O	O
anabolism	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
adipocyte	NN	O	B-protein
fatty	NN	O	I-protein
acid	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
aP2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
a	NN	O	O
key	NN	O	O
regulator	NN	O	O
of	NN	O	O
adipocyte	NN	O	B-cell_type
differentiation	NN	O	O
.	NN	O	O

PPARgamma	NN	O	B-protein
is	NN	O	O
also	NN	O	O
expressed	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
up-regulated	NN	O	O
in	NN	O	O
activated	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
PPARgamma	NN	O	B-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
differentiation	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
to	NN	O	O
foam	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
atherosclerotic	NN	O	O
lesions	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
both	NN	O	O
natural	NN	O	O
and	NN	O	O
synthetic	NN	O	O
PPARgamma	NN	O	O
agonists	NN	O	O
induce	NN	O	O
time-	NN	O	O
and	NN	O	O
dose-dependent	NN	O	O
increases	NN	O	O
in	NN	O	O
aP2	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
both	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
PPARgamma	NN	O	B-protein
activation	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
monocyte	NN	O	B-cell_type
differentiation	NN	O	O
and	NN	O	O
function	NN	O	O
analogous	NN	O	O
to	NN	O	O
its	NN	O	O
well-characterized	NN	O	O
role	NN	O	O
in	NN	O	O
adipocytes	NN	O	B-cell_type
.	NN	O	I-cell_type

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
physical	NN	O	O
interaction	NN	O	O
between	NN	O	O
calcineurin	NN	O	B-protein
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFATp	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
calcium/calmodulin-dependent	NN	O	B-protein
serine/threonine	NN	O	I-protein
phosphatase	NN	O	I-protein
,	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
plays	NN	O	O
a	NN	O	O
pivotal	NN	O	O
role	NN	O	O
in	NN	O	O
transducing	NN	O	O
membrane-associated	NN	O	O
signals	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
putative	NN	O	O
targets	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
is	NN	O	O
the	NN	O	O
pre-existing	NN	O	O
,	NN	O	O
cytosolic	NN	O	B-protein
component	NN	O	I-protein
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFATp	NN	O	B-protein
;	NN	O	O
also	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
NFAT1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
required	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
by	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
drugs	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
prevents	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	O
its	NN	O	O
translocation	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
physical	NN	O	O
interaction	NN	O	O
between	NN	O	O
calcineurin	NN	O	B-protein
and	NN	O	O
NFATp	NN	O	B-protein
has	NN	O	O
not	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
from	NN	O	O
lysates	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
immobilized	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
induced	NN	O	O
a	NN	O	O
shift	NN	O	O
in	NN	O	O
the	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
that	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
its	NN	O	O
dephosphorylation	NN	O	O
.	NN	O	O

This	NN	O	O
dephosphorylation	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
treatment	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
or	NN	O	O
FK506	NN	O	O
prior	NN	O	O
to	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Of	NN	O	O
note	NN	O	O
,	NN	O	O
both	NN	O	O
the	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
the	NN	O	O
dephosphorylated	NN	O	O
form	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
both	NN	O	O
forms	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
to	NN	O	O
calcineurin	NN	O	B-protein
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
with	NN	O	O
a	NN	O	O
complex	NN	O	O
of	NN	O	O
FK506	NN	O	O
and	NN	O	O
its	NN	O	O
ligand	NN	O	O
FKBP12	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
these	NN	O	O
data	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
a	NN	O	O
direct	NN	O	O
interaction	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
with	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	O
that	NN	O	O
this	NN	O	O
interaction	NN	O	O
does	NN	O	O
not	NN	O	O
depend	NN	O	O
upon	NN	O	O
the	NN	O	O
phosphorylation	NN	O	B-DNA
sites	NN	O	I-DNA
of	NN	O	O
NFATp	NN	O	B-protein
affected	NN	O	O
by	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cross-linking	NN	O	O
CD40	NN	O	B-protein
on	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
preferentially	NN	O	O
induces	NN	O	O
stress-activated	NN	O	B-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
rather	NN	O	O
than	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

The	NN	O	O
B	NN	O	B-protein
cell-associated	NN	O	I-protein
surface	NN	O	I-protein
molecule	NN	O	I-protein
CD40	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
T	NN	O	O
cell-dependent	NN	O	O
B	NN	O	O
cell	NN	O	O
maturation	NN	O	O
,	NN	O	O
as	NN	O	O
individuals	NN	O	O
with	NN	O	O
defects	NN	O	O
in	NN	O	O
either	NN	O	O
CD40	NN	O	B-protein
or	NN	O	O
its	NN	O	O
ligand	NN	O	O
are	NN	O	O
impaired	NN	O	O
in	NN	O	O
immunoglobulin	NN	O	B-protein
isotype	NN	O	O
class	NN	O	O
switching	NN	O	O
and	NN	O	O
germinal	NN	O	O
center	NN	O	O
formation	NN	O	O
.	NN	O	O

CD40	NN	O	B-protein
signaling	NN	O	O
activates	NN	O	O
downstream	NN	O	B-protein
effectors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
Lyn	NN	O	B-protein
,	NN	O	O
the	NN	O	O
phosphatidylinositol-3-kinase	NN	O	B-protein
(	NN	O	O
PI-3	NN	O	B-protein
kinase	NN	O	I-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
stress-activated	NN	O	B-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
(	NN	O	O
SAPK	NN	O	B-protein
)	NN	O	O
are	NN	O	O
activated	NN	O	O
after	NN	O	O
CD40	NN	O	B-protein
cross-linking	NN	O	O
on	NN	O	O
various	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
or	NN	O	O
human	NN	O	B-cell_type
tonsillar	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
activation	NN	O	O
is	NN	O	O
rapid	NN	O	O
and	NN	O	O
transient	NN	O	O
and	NN	O	O
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
a	NN	O	O
cyclosporin	NN	O	O
A-insensitive	NN	O	O
pathway	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
this	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
appears	NN	O	O
not	NN	O	O
to	NN	O	O
rely	NN	O	O
on	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
.	NN	O	O

While	NN	O	O
CD40	NN	O	B-protein
ligation	NN	O	O
strongly	NN	O	O
activates	NN	O	O
the	NN	O	O
SAPKs	NN	O	B-protein
(	NN	O	O
up	NN	O	O
to	NN	O	O
25-fold	NN	O	O
)	NN	O	O
,	NN	O	O
it	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
family	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
;	NN	O	O
ERK1	NN	O	B-protein
and	NN	O	O
ERK2	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
these	NN	O	O
data	NN	O	O
,	NN	O	O
CD40	NN	O	B-protein
signals	NN	O	O
up-regulate	NN	O	O
c-jun	NN	O	B-RNA
but	NN	O	I-RNA
not	NN	O	I-RNA
c-fos	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
alter	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
ATF2	NN	O	B-protein
but	NN	O	O
not	NN	O	O
the	NN	O	O
Raf-1	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
preferentially	NN	O	O
induces	NN	O	O
SAPK	NN	O	B-protein
but	NN	O	O
not	NN	O	O
MAPK	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor-CD3	NN	O	I-protein
complex	NN	O	I-protein
signaling	NN	O	O
pathway	NN	O	O
by	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
phosphatase	NN	O	I-protein
inhibitor	NN	O	O
pervanadate	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
CD45	NN	O	B-protein
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
two	NN	O	O
interconnected	NN	O	O
Lck/Fyn-	NN	O	O
or	NN	O	O
zap-70-dependent	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

The	NN	O	O
tyrosine	NN	O	B-protein
phosphatase	NN	O	I-protein
specific	NN	O	O
inhibitor	NN	O	O
pervanadate	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
activator	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
through	NN	O	O
induction	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
phosphorylation	NN	O	I-protein
and	NN	O	O
downstream	NN	O	O
events	NN	O	O
of	NN	O	O
the	NN	O	O
activation	NN	O	O
cascade	NN	O	O
.	NN	O	O

Using	NN	O	O
CD45-	NN	O	B-cell_line
or	NN	O	I-cell_line
CD3-negative	NN	O	I-cell_line
variants	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
different	NN	O	O
biochemical	NN	O	O
events	NN	O	O
induced	NN	O	O
by	NN	O	O
pervanadate	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
at	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
of	NN	O	O
either	NN	O	O
CD45	NN	O	B-protein
or	NN	O	O
CD3	NN	O	B-protein
.	NN	O	O

CD45	NN	O	B-protein
-dependent	NN	O	O
events	NN	O	O
such	NN	O	O
as	NN	O	O
tyrosine	NN	O	B-protein
phosphorylation	NN	O	I-protein
of	NN	O	O
Shc	NN	O	B-protein
,	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
stimulation	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
of	NN	O	O
CD69	NN	O	B-protein
and	NN	O	O
CD25	NN	O	B-protein
surface	NN	O	O
expression	NN	O	O
paralleled	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
lck	NN	O	B-protein
and	NN	O	O
fyn	NN	O	B-protein
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
stimulation	NN	O	O
of	NN	O	O
calcium	NN	O	O
influx	NN	O	O
,	NN	O	O
a	NN	O	O
CD3	NN	O	B-protein
-dependent	NN	O	O
event	NN	O	O
,	NN	O	O
paralleled	NN	O	O
zap-70	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
T-cell	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
-CD3	NN	O	B-protein
(	NN	O	I-protein
TcR-CD3	NN	O	I-protein
)	NN	O	I-protein
complex	NN	O	I-protein
is	NN	O	O
functionally	NN	O	O
linked	NN	O	O
to	NN	O	O
two	NN	O	O
different	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
PTK	NN	O	B-protein
)	NN	O	O
modules	NN	O	O
with	NN	O	O
separate	NN	O	O
specific	NN	O	O
functions	NN	O	O
and	NN	O	O
that	NN	O	O
CD45	NN	O	B-protein
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
regulator	NN	O	O
of	NN	O	O
this	NN	O	O
coupling	NN	O	O
.	NN	O	O

-DOCSTART-	O

Vitamin	NN	O	O
D	NN	O	O
analogs	NN	O	O
,	NN	O	O
20-Epi-22-oxa-24a	NN	O	O
,	NN	O	O
26a	NN	O	O
,	NN	O	O
27a	NN	O	O
,	NN	O	O
-trihomo-1alpha	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2-vitamin	NN	O	O
D3	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
24	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2-22-ene-24-cyclopropyl-vitamin	NN	O	O
D3	NN	O	O
and	NN	O	O
1alpha	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2-lumisterol3	NN	O	O
prime	NN	O	O
NB4	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
for	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
via	NN	O	O
nongenomic	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
,	NN	O	O
involving	NN	O	O
calcium	NN	O	O
and	NN	O	O
calpain	NN	O	O
.	NN	O	O

Side-chain	NN	O	O
modified	NN	O	O
vitamin	NN	O	O
D	NN	O	O
analogs	NN	O	O
including	NN	O	O
20-Epi-22-oxa-24a	NN	O	O
,	NN	O	O
26a	NN	O	O
,	NN	O	O
27a-trihomo-1alpha	NN	O	O
,	NN	O	O
2	NN	O	O
5-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
KH1060	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
1	NN	O	O
,	NN	O	O
24-dihydroxy-22-ene-24-cyclopropyl-vitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
MC903	NN	O	O
)	NN	O	O
were	NN	O	O
originally	NN	O	O
designed	NN	O	O
to	NN	O	O
aid	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
hyperproliferative	NN	O	O
disorders	NN	O	O
including	NN	O	O
psoriasis	NN	O	O
and	NN	O	O
cancer	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
these	NN	O	O
analogs	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
6-cis-locked	NN	O	O
conformer	NN	O	O
,	NN	O	O
1alpha	NN	O	B-cell_line
,	NN	O	I-cell_line
25-dihydroxy-lumisterol3	NN	O	I-cell_line
(	NN	O	I-cell_line
JN	NN	O	I-cell_line
)	NN	O	I-cell_line
prime	NN	O	I-cell_line
NB4	NN	O	I-cell_line
cells	NN	O	I-cell_line
for	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
MC903	NN	O	O
and	NN	O	O
KH1060	NN	O	O
was	NN	O	O
presumed	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
vitamin	NN	O	I-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
VDRnuc	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Differentiation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
all	NN	O	O
analogs	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
inhibited	NN	O	O
by	NN	O	O
1beta	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
HL	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
antagonist	NN	O	O
to	NN	O	O
the	NN	O	O
nongenomic	NN	O	O
activities	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25D3	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
although	NN	O	O
MC903	NN	O	O
and	NN	O	O
KH1060	NN	O	O
may	NN	O	O
bind	NN	O	O
the	NN	O	O
VDRnuc	NN	O	B-protein
,	NN	O	O
that	NN	O	O
the	NN	O	O
differentiative	NN	O	O
activities	NN	O	O
of	NN	O	O
these	NN	O	O
agents	NN	O	O
requires	NN	O	O
nongenomic	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
1alpha	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2-d5-previtamin	NN	O	O
D3	NN	O	O
(	NN	O	O
HF	NN	O	O
)	NN	O	O
,	NN	O	O
JN	NN	O	O
,	NN	O	O
KH1060	NN	O	O
,	NN	O	O
and	NN	O	O
MC903	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
PKC	NN	O	B-protein
delta	NN	O	I-protein
and	NN	O	O
translocation	NN	O	O
of	NN	O	O
both	NN	O	O
isoforms	NN	O	O
to	NN	O	O
the	NN	O	O
particulate	NN	O	O
fraction	NN	O	O
,	NN	O	O
and	NN	O	O
PKC	NN	O	B-protein
alpha	NN	O	I-protein
to	NN	O	O
the	NN	O	O
nuclear	NN	O	O
fraction	NN	O	O
.	NN	O	O

The	NN	O	O
full	NN	O	O
differentiation	NN	O	O
response	NN	O	O
with	NN	O	O
combinations	NN	O	O
of	NN	O	O
analogs	NN	O	O
and	NN	O	O
TPA	NN	O	O
was	NN	O	O
inhibited	NN	O	O
50	NN	O	O
%	NN	O	O
by	NN	O	O
the	NN	O	O
membrane	NN	O	O
permeable	NN	O	O
Ca2+	NN	O	O
chelator	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
2-bis	NN	O	O
(	NN	O	O
o-aminophenoxy	NN	O	O
)	NN	O	O
-ethane-N	NN	O	O
,	NN	O	O
N	NN	O	O
,	NN	O	O
N	NN	O	O
'	NN	O	O
,	NN	O	O
N'-tetraacetic	NN	O	O
acid	NN	O	O
(	NN	O	O
BAPTA-AM	NN	O	O
)	NN	O	O
or	NN	O	O
calpain	NN	O	B-protein
inhibitor	NN	O	O
I	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
intracellular	NN	O	O
free	NN	O	O
calcium	NN	O	O
and	NN	O	O
the	NN	O	O
calcium-dependent	NN	O	B-protein
protease	NN	O	I-protein
,	NN	O	O
calpain	NN	O	B-protein
play	NN	O	O
critical	NN	O	O
roles	NN	O	O
in	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Intracellular	NN	O	O
calcium	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
most	NN	O	O
critical	NN	O	O
in	NN	O	O
the	NN	O	O
1	NN	O	O
,	NN	O	O
25D3-priming	NN	O	O
stage	NN	O	O
of	NN	O	O
differentiation	NN	O	O
,	NN	O	O
while	NN	O	O
calpain	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
in	NN	O	O
the	NN	O	O
TPA	NN	O	O
maturation	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Neutrophil	NN	O	B-cell_type
maturation	NN	O	O
and	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
.	NN	O	O

Neutrophil	NN	O	B-cell_type
maturation	NN	O	O
occurs	NN	O	O
in	NN	O	O
well	NN	O	O
defined	NN	O	O
morphological	NN	O	O
stages	NN	O	O
that	NN	O	O
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
acquisition	NN	O	O
of	NN	O	O
molecular	NN	O	B-protein
markers	NN	O	I-protein
associated	NN	O	O
with	NN	O	O
neutrophil	NN	O	O
function	NN	O	O
.	NN	O	O

A	NN	O	O
variety	NN	O	O
of	NN	O	O
factors	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
terminal	NN	O	O
neutrophil	NN	O	B-cell_type
maturation	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
vitamin	NN	O	O
A	NN	O	O
derivative	NN	O	O
,	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
.	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
can	NN	O	O
directly	NN	O	O
modulate	NN	O	O
gene	NN	O	O
expression	NN	O	O
via	NN	O	O
binding	NN	O	O
to	NN	O	O
its	NN	O	O
nuclear	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
which	NN	O	O
can	NN	O	O
,	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
role	NN	O	O
for	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
during	NN	O	O
neutrophil	NN	O	B-cell_type
maturation	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
from	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
sources.	NN	O	O
Here	NN	O	O
we	NN	O	O
present	NN	O	O
a	NN	O	O
review	NN	O	O
of	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
action	NN	O	O
and	NN	O	O
the	NN	O	O
major	NN	O	O
evidence	NN	O	O
showing	NN	O	O
that	NN	O	O
normal	NN	O	O
retinoid	NN	O	O
signaling	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
neutrophil	NN	O	B-cell_type
maturation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-DNA
T-cell	NN	O	I-DNA
lymphotrophic	NN	O	I-DNA
virus	NN	O	I-DNA
type-I	NN	O	I-DNA
tax	NN	O	I-DNA
gene	NN	O	I-DNA
induces	NN	O	O
secretion	NN	O	O
of	NN	O	O
human	NN	O	B-protein
macrophage	NN	O	I-protein
inflammatory	NN	O	I-protein
protein-1alpha	NN	O	I-protein
.	NN	O	O

Human	NN	O	O
T-cell	NN	O	O
lymphotropic	NN	O	O
virus	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
encodes	NN	O	O
for	NN	O	O
a	NN	O	O
40-kDa	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
Tax	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
immortalization	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Tax	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
transactivate	NN	O	O
several	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
MIP-1alpha	NN	O	B-protein
is	NN	O	O
selectively	NN	O	O
expressed	NN	O	O
and	NN	O	O
secreted	NN	O	O
in	NN	O	O
the	NN	O	O
tax	NN	O	B-cell_line
transfected	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
upon	NN	O	O
mitogen	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
MIP-1alpha-R	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
these	NN	O	O
cells	NN	O	O
suggests	NN	O	O
an	NN	O	O
autocrine	NN	O	O
role	NN	O	O
for	NN	O	O
this	NN	O	O
chemokine	NN	O	B-protein
in	NN	O	O
HTLV-I	NN	O	B-cell_type
infected	NN	O	I-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

Induced	NN	O	O
MIP-1alpha	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
secretion	NN	O	O
in	NN	O	O
PMA/PHA	NN	O	B-cell_line
stimulated	NN	O	I-cell_line
tax	NN	O	I-cell_line
transfected	NN	O	I-cell_line
cells	NN	O	I-cell_line
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
noninduction	NN	O	O
of	NN	O	O
MNP-1	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
intimately	NN	O	O
involved	NN	O	O
in	NN	O	O
downmodulating	NN	O	O
the	NN	O	O
MIP-1alpha	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
.	NN	O	O

Rheumatoid	NN	O	O
arthritis	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
chronic	NN	O	O
autoimmune	NN	O	O
inflammatory	NN	O	O
disease	NN	O	O
in	NN	O	O
which	NN	O	O
unknown	NN	O	O
arthrogenic	NN	O	B-protein
autoantigen	NN	O	I-protein
is	NN	O	O
presented	NN	O	O
to	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
strong	NN	O	O
association	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
with	NN	O	O
an	NN	O	O
epitope	NN	O	B-protein
within	NN	O	O
the	NN	O	O
HLA-DR	NN	O	B-protein
chain	NN	O	I-protein
shared	NN	O	O
between	NN	O	O
various	NN	O	O
alleles	NN	O	O
of	NN	O	O
HLA-DR4	NN	O	B-protein
and	NN	O	O
DR1	NN	O	B-protein
emphasizes	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
antigen	NN	O	O
presentation	NN	O	O
.	NN	O	O

This	NN	O	O
immune	NN	O	O
response	NN	O	O
predominantly	NN	O	O
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
synovial	NN	O	O
tissue	NN	O	O
and	NN	O	O
fluid	NN	O	O
of	NN	O	O
the	NN	O	O
joints	NN	O	O
and	NN	O	O
autoreactive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
readily	NN	O	O
demonstrable	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
synovial	NN	O	O
compartment	NN	O	O
and	NN	O	O
blood	NN	O	O
.	NN	O	O

Circulating	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
DC	NN	O	B-cell_type
)	NN	O	O
are	NN	O	O
phenotypically	NN	O	O
and	NN	O	O
functionally	NN	O	O
identical	NN	O	O
with	NN	O	O
normal	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
(	NN	O	I-cell_type
PB	NN	O	I-cell_type
)	NN	O	I-cell_type
DC	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
synovial	NN	O	O
tissue	NN	O	O
,	NN	O	O
fully	NN	O	O
differentiated	NN	O	O
perivascular	NN	O	B-cell_type
DC	NN	O	I-cell_type
are	NN	O	O
found	NN	O	O
in	NN	O	O
close	NN	O	O
association	NN	O	O
with	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
with	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
follicles	NN	O	I-cell_type
,	NN	O	O
sometimes	NN	O	O
containing	NN	O	O
follicular	NN	O	B-cell_type
DC	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
perivascular	NN	O	B-cell_type
DC	NN	O	I-cell_type
migrate	NN	O	O
across	NN	O	O
the	NN	O	O
activated	NN	O	O
endothelium	NN	O	O
from	NN	O	O
blood	NN	O	O
and	NN	O	O
receive	NN	O	O
differentiative	NN	O	O
signals	NN	O	O
within	NN	O	O
the	NN	O	O
joint	NN	O	O
from	NN	O	O
monocyte-derived	NN	O	B-protein
cytokines	NN	O	I-protein
and	NN	O	O
CD40-ligand+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
SF	NN	O	O
,	NN	O	O
DC	NN	O	B-cell_type
manifest	NN	O	O
an	NN	O	O
intermediate	NN	O	O
phenotype	NN	O	O
,	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
monocyte-derived	NN	O	B-cell_type
DC	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Like	NN	O	O
a	NN	O	O
delayed-type	NN	O	O
hypersensitivity	NN	O	O
response	NN	O	O
,	NN	O	O
the	NN	O	O
rheumatoid	NN	O	O
synovium	NN	O	O
represents	NN	O	O
an	NN	O	O
effector	NN	O	O
site	NN	O	O
.	NN	O	O

DC	NN	O	O
at	NN	O	O
many	NN	O	O
effector	NN	O	O
sites	NN	O	O
have	NN	O	O
a	NN	O	O
characteristic	NN	O	O
pattern	NN	O	O
of	NN	O	O
infiltration	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
important	NN	O	O
to	NN	O	O
note	NN	O	O
that	NN	O	O
the	NN	O	O
effector	NN	O	O
response	NN	O	O
is	NN	O	O
not	NN	O	O
self-limiting	NN	O	O
in	NN	O	O
RA	NN	O	O
autoimmune	NN	O	O
inflammation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
article	NN	O	O
,	NN	O	O
we	NN	O	O
argue	NN	O	O
that	NN	O	O
the	NN	O	O
presentation	NN	O	O
of	NN	O	O
self-antigen	NN	O	B-protein
by	NN	O	O
DC	NN	O	B-cell_type
and	NN	O	O
by	NN	O	O
autoantibody-producing	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
perpetuation	NN	O	O
of	NN	O	O
the	NN	O	O
autoimmune	NN	O	O
response	NN	O	O
.	NN	O	O

Permanently	NN	O	O
arresting	NN	O	O
this	NN	O	O
ongoing	NN	O	O
immune	NN	O	O
response	NN	O	O
with	NN	O	O
either	NN	O	O
pharmaceutical	NN	O	O
agents	NN	O	O
or	NN	O	O
immunotherapy	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
challenge	NN	O	O
for	NN	O	O
immunology	NN	O	O
.	NN	O	O

-DOCSTART-	O

Thymocyte-thymic	NN	O	O
epithelial	NN	O	B-cell_type
cell	NN	O	I-cell_type
interaction	NN	O	O
leads	NN	O	O
to	NN	O	O
high-level	NN	O	O
replication	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
exclusively	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
CD3	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
:	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
for	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
interleukin-7	NN	O	B-protein
.	NN	O	O

This	NN	O	O
work	NN	O	O
aims	NN	O	O
at	NN	O	O
identifying	NN	O	O
the	NN	O	O
thymocyte	NN	O	O
subpopulation	NN	O	O
able	NN	O	O
to	NN	O	O
support	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
replication	NN	O	O
under	NN	O	O
the	NN	O	O
biological	NN	O	O
stimuli	NN	O	O
of	NN	O	O
the	NN	O	O
thymic	NN	O	O
microenvironment	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
interaction	NN	O	O
with	NN	O	O
thymic	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
TEC	NN	O	B-cell_type
)	NN	O	O
induces	NN	O	O
a	NN	O	O
high-level	NN	O	O
replication	NN	O	O
of	NN	O	O
the	NN	O	O
T-tropic	NN	O	B-protein
primary	NN	O	I-protein
isolate	NN	O	I-protein
HIV-1	NN	O	I-protein
(	NN	O	O
B-LAIp	NN	O	B-protein
)	NN	O	O
exclusively	NN	O	O
in	NN	O	O
the	NN	O	O
mature	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
CD3	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interleukin-7	NN	O	B-protein
(	NN	O	O
IL-7	NN	O	B-protein
)	NN	O	O
,	NN	O	O
secreted	NN	O	O
during	NN	O	O
this	NN	O	O
interaction	NN	O	O
,	NN	O	O
are	NN	O	O
critical	NN	O	O
cytokines	NN	O	O
for	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
transactivation	NN	O	O
through	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
.	NN	O	O

TNF	NN	O	B-protein
is	NN	O	O
the	NN	O	O
major	NN	O	O
inducer	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
particularly	NN	O	O
of	NN	O	O
the	NN	O	O
p50-p65	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
IL-7	NN	O	B-protein
acts	NN	O	O
as	NN	O	O
a	NN	O	O
cofactor	NN	O	O
by	NN	O	O
sustaining	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
p75	NN	O	B-protein
TNF	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
requirement	NN	O	O
for	NN	O	O
TNF	NN	O	B-protein
is	NN	O	O
further	NN	O	O
confirmed	NN	O	O
by	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
the	NN	O	O
inability	NN	O	O
of	NN	O	O
the	NN	O	O
intermediate	NN	O	B-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
CD3	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
to	NN	O	O
replicate	NN	O	O
the	NN	O	O
virus	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
defect	NN	O	O
in	NN	O	O
TNF	NN	O	B-protein
production	NN	O	O
during	NN	O	O
their	NN	O	O
interaction	NN	O	O
with	NN	O	O
TEC	NN	O	B-cell_type
and	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
nuclear	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
these	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
exogenous	NN	O	O
TNF	NN	O	B-protein
to	NN	O	O
the	NN	O	O
intermediate	NN	O	B-cell_line
thymocyte	NN	O	I-cell_line
cultures	NN	O	I-cell_line
induces	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
and	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
promote	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
the	NN	O	O
cocultures	NN	O	O
with	NN	O	O
TEC	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
other	NN	O	O
major	NN	O	O
subpopulation	NN	O	O
expressing	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
namely	NN	O	O
,	NN	O	O
the	NN	O	O
double-positive	NN	O	B-cell_type
(	NN	O	I-cell_type
DP	NN	O	I-cell_type
)	NN	O	I-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CD3	NN	O	I-cell_type
(	NN	O	I-cell_type
+/-	NN	O	I-cell_type
)	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
,	NN	O	O
despite	NN	O	O
the	NN	O	O
entry	NN	O	O
of	NN	O	O
the	NN	O	O
virus	NN	O	O
,	NN	O	O
do	NN	O	O
not	NN	O	O
produce	NN	O	O
a	NN	O	O
significant	NN	O	O
level	NN	O	O
of	NN	O	O
virus	NN	O	O
,	NN	O	O
presumably	NN	O	O
because	NN	O	O
they	NN	O	O
are	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
TNF	NN	O	B-protein
and	NN	O	O
IL-7	NN	O	B-protein
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
despite	NN	O	O
an	NN	O	O
efficient	NN	O	O
entry	NN	O	O
of	NN	O	O
the	NN	O	O
virus	NN	O	O
in	NN	O	O
all	NN	O	O
the	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
subpopulations	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
high	NN	O	O
viral	NN	O	O
load	NN	O	O
may	NN	O	O
be	NN	O	O
generated	NN	O	O
exclusively	NN	O	O
within	NN	O	O
the	NN	O	O
mature	NN	O	B-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
CD3	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
subset	NN	O	I-cell_type
of	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
under	NN	O	O
conditions	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
after	NN	O	O
infection	NN	O	O
,	NN	O	O
TNF	NN	O	B-protein
might	NN	O	O
also	NN	O	O
be	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
intermediate	NN	O	B-cell_type
thymocyte	NN	O	I-cell_type
compartment	NN	O	I-cell_type
,	NN	O	O
leading	NN	O	O
to	NN	O	O
efficient	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappaB-mediated	NN	O	B-cell_type
up-regulation	NN	O	I-cell_type
of	NN	O	O
Bcl-x	NN	O	B-protein
and	NN	O	O
Bfl-1/A1	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
CD40	NN	O	B-protein
survival	NN	O	O
signaling	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
thymus-dependent	NN	O	O
humoral	NN	O	O
immune	NN	O	O
responses	NN	O	O
and	NN	O	O
rescuing	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Many	NN	O	O
of	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
are	NN	O	O
believed	NN	O	O
to	NN	O	O
be	NN	O	O
achieved	NN	O	O
through	NN	O	O
altered	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
Bcl-x	NN	O	B-protein
,	NN	O	O
a	NN	O	O
known	NN	O	O
CD40-regulated	NN	O	B-protein
antiapoptotic	NN	O	I-protein
molecule	NN	O	I-protein
,	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
related	NN	O	O
antiapoptotic	NN	O	B-protein
molecule	NN	O	I-protein
,	NN	O	O
A1/Bfl-1	NN	O	B-protein
,	NN	O	O
as	NN	O	O
a	NN	O	O
CD40-inducible	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
pathway	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
a	NN	O	O
dominant-active	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
abolished	NN	O	O
CD40	NN	O	B-protein
-induced	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
Bfl-1	NN	O	B-protein
and	NN	O	O
Bcl-x	NN	O	B-protein
genes	NN	O	O
and	NN	O	O
also	NN	O	O
eliminated	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
to	NN	O	O
rescue	NN	O	O
Fas	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Within	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
Bcl-x	NN	O	B-protein
,	NN	O	O
a	NN	O	O
potential	NN	O	O
NF-kappaB-binding	NN	O	B-DNA
sequence	NN	O	I-DNA
was	NN	O	O
found	NN	O	O
to	NN	O	O
support	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
physiological	NN	O	O
levels	NN	O	O
of	NN	O	O
Bcl-x	NN	O	B-cell_type
protected	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
Fas-mediated	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
cell	NN	O	O
survival	NN	O	O
proceeds	NN	O	O
through	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Stimulation	NN	O	O
of	NN	O	O
neutrophil	NN	O	B-cell_type
interleukin-8	NN	O	B-protein
production	NN	O	O
by	NN	O	O
eosinophil	NN	O	B-protein
granule	NN	O	I-protein
major	NN	O	I-protein
basic	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
evaluated	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
eosinophil	NN	O	B-protein
granule	NN	O	I-protein
major	NN	O	I-protein
basic	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
MBP	NN	O	B-protein
)	NN	O	O
to	NN	O	O
stimulate	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-8	NN	O	I-protein
production	NN	O	O
by	NN	O	O
neutrophils	NN	O	B-cell_type
.	NN	O	O

MBP	NN	O	B-protein
over	NN	O	O
the	NN	O	O
concentration	NN	O	O
range	NN	O	O
of	NN	O	O
0.1	NN	O	O
to	NN	O	O
10	NN	O	O
microM	NN	O	O
stimulated	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
up	NN	O	O
to	NN	O	O
approximately	NN	O	O
8	NN	O	O
ng/ml	NN	O	O
IL-8	NN	O	B-protein
.	NN	O	O

Incubation	NN	O	O
with	NN	O	O
2	NN	O	O
microM	NN	O	O
MBP	NN	O	B-protein
showed	NN	O	O
that	NN	O	O
,	NN	O	O
after	NN	O	O
a	NN	O	O
1	NN	O	O
h	NN	O	O
lag	NN	O	O
,	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
release	NN	O	O
increased	NN	O	O
with	NN	O	O
time	NN	O	O
for	NN	O	O
approximately	NN	O	O
10	NN	O	O
h	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
2	NN	O	O
microM	NN	O	O
concentration	NN	O	O
,	NN	O	O
eosinophil	NN	O	B-protein
cationic	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
eosinophil-derived	NN	O	O
neurotoxin	NN	O	O
,	NN	O	O
and	NN	O	O
eosinophil	NN	O	B-protein
peroxidase	NN	O	I-protein
did	NN	O	O
not	NN	O	O
stimulate	NN	O	O
significant	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
production	NN	O	O
.	NN	O	O

MBP	NN	O	B-protein
stimulated	NN	O	O
2-fold	NN	O	O
increases	NN	O	O
in	NN	O	O
IL-8	NN	O	B-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
(	NN	O	O
mRNA	NN	O	B-RNA
)	NN	O	O
after	NN	O	O
1	NN	O	O
and	NN	O	O
3	NN	O	O
h	NN	O	O
of	NN	O	O
incubation	NN	O	O
,	NN	O	O
which	NN	O	O
were	NN	O	O
blocked	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
stimulation	NN	O	O
with	NN	O	O
MBP	NN	O	B-protein
did	NN	O	O
not	NN	O	O
produce	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappaB	NN	O	I-protein
or	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
.	NN	O	O

No	NN	O	O
NF-IL-6	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
stimulation	NN	O	O
with	NN	O	O
MBP	NN	O	B-protein
prolonged	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
IL-8	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

MBP	NN	O	B-protein
also	NN	O	O
induced	NN	O	O
transient	NN	O	O
increases	NN	O	O
in	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
macrophage	NN	O	B-protein
inflammatory	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MIP	NN	O	I-protein
)	NN	O	I-protein
-1alpha	NN	O	I-protein
and	NN	O	O
MIP-1beta	NN	O	B-protein
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
stimulate	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
either	NN	O	O
chemokine	NN	O	B-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
MBP	NN	O	B-protein
is	NN	O	O
selective	NN	O	O
among	NN	O	O
the	NN	O	O
eosinophil	NN	O	B-protein
granule	NN	O	I-protein
proteins	NN	O	I-protein
as	NN	O	O
a	NN	O	O
stimulus	NN	O	O
for	NN	O	O
neutrophil	NN	O	B-cell_type
IL-8	NN	O	B-protein
release	NN	O	O
and	NN	O	O
,	NN	O	O
further	NN	O	O
,	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
neutrophil	NN	O	B-cell_type
IL-8	NN	O	B-protein
release	NN	O	O
by	NN	O	O
MBP	NN	O	B-protein
involves	NN	O	O
both	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
posttranscriptional	NN	O	O
regulation	NN	O	O
.	NN	O	O

We	NN	O	O
postulate	NN	O	O
that	NN	O	O
MBP	NN	O	B-protein
-induced	NN	O	O
release	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
by	NN	O	O
neutrophils	NN	O	B-cell_type
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
acute	NN	O	O
asthma	NN	O	O
and	NN	O	O
other	NN	O	O
inflammatory	NN	O	O
lung	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
TCL1	NN	O	B-DNA
oncogene	NN	O	I-DNA
,	NN	O	O
or	NN	O	O
transcripts	NN	O	O
from	NN	O	O
its	NN	O	O
homologue	NN	O	O
MTCP1/c6.1B	NN	O	B-DNA
,	NN	O	O
in	NN	O	O
leukaemic	NN	O	O
and	NN	O	O
non-leukaemic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
ataxia	NN	O	O
telangiectasia	NN	O	O
patients	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
the	NN	O	O
recessively	NN	O	O
inherited	NN	O	O
disorder	NN	O	O
ataxia	NN	O	O
telangiectasia	NN	O	O
(	NN	O	O
A-T	NN	O	O
)	NN	O	O
have	NN	O	O
a	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
specific	NN	O	O
chromosome	NN	O	B-DNA
translocations	NN	O	I-DNA
which	NN	O	O
can	NN	O	O
be	NN	O	O
easily	NN	O	O
observed	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
show	NN	O	O
a	NN	O	O
greatly	NN	O	O
increased	NN	O	O
predisposition	NN	O	O
to	NN	O	O
leukaemia/lymphoma	NN	O	O
,	NN	O	O
mainly	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
origin	NN	O	O
.	NN	O	O

Some	NN	O	O
translocation	NN	O	B-cell_type
cells	NN	O	I-cell_type
proliferate	NN	O	O
into	NN	O	O
a	NN	O	O
large	NN	O	O
clone	NN	O	O
and	NN	O	O
may	NN	O	O
develop	NN	O	O
into	NN	O	O
T	NN	O	O
cell	NN	O	O
prolymphocytic	NN	O	O
leukaemia	NN	O	O
(	NN	O	O
T-PLL	NN	O	O
)	NN	O	O
.	NN	O	O

By	NN	O	O
the	NN	O	O
time	NN	O	O
of	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
T-PLL	NN	O	O
,	NN	O	O
the	NN	O	O
clone	NN	O	O
contains	NN	O	O
many	NN	O	O
more	NN	O	O
genetic	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
form	NN	O	O
of	NN	O	O
additional	NN	O	O
translocations	NN	O	O
.	NN	O	O

T-PLL	NN	O	O
is	NN	O	O
also	NN	O	O
seen	NN	O	O
in	NN	O	O
non-A-T	NN	O	O
individuals	NN	O	O
where	NN	O	O
expression	NN	O	O
of	NN	O	O
either	NN	O	O
TCL1	NN	O	B-protein
(	NN	O	O
at	NN	O	O
14q32	NN	O	B-DNA
)	NN	O	O
or	NN	O	O
the	NN	O	O
c6.1B/MTCP1	NN	O	B-protein
A1	NN	O	I-protein
transcript	NN	O	I-protein
(	NN	O	O
at-Xq28	NN	O	B-DNA
)	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
just	NN	O	O
a	NN	O	O
few	NN	O	O
instances	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
,	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
TCL1	NN	O	B-protein
occurs	NN	O	O
in	NN	O	O
leukaemic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
A-T	NN	O	O
patients	NN	O	O
with	NN	O	O
chromosome	NN	O	B-DNA
14	NN	O	I-DNA
rearrangements	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
TCL1	NN	O	B-protein
also	NN	O	O
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
preleukaemic	NN	O	B-cell_type
clone	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
A-T	NN	O	O
patients	NN	O	O
containing	NN	O	O
the	NN	O	O
primary	NN	O	O
translocation	NN	O	O
alone	NN	O	O
.	NN	O	O

Some	NN	O	O
expression	NN	O	O
of	NN	O	O
TCL1	NN	O	B-protein
could	NN	O	O
also	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
randomly	NN	O	O
selected	NN	O	O
A-T	NN	O	O
patients	NN	O	O
without	NN	O	O
large	NN	O	O
cytogenetic	NN	O	O
clones	NN	O	O
and	NN	O	O
without	NN	O	O
any	NN	O	O
evidence	NN	O	O
of	NN	O	O
leukaemic	NN	O	O
change	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
B1	NN	O	B-RNA
transcript	NN	O	I-RNA
from	NN	O	O
a	NN	O	O
second	NN	O	O
gene	NN	O	O
,	NN	O	O
MTCP1	NN	O	B-DNA
,	NN	O	O
occurred	NN	O	O
at	NN	O	O
a	NN	O	O
relatively	NN	O	O
high	NN	O	O
level	NN	O	O
only	NN	O	O
in	NN	O	O
two	NN	O	O
T-PLL	NN	O	O
tumours	NN	O	O
from	NN	O	O
A-T	NN	O	O
patients	NN	O	O
with	NN	O	O
t	NN	O	O
(	NN	O	O
X	NN	O	O
;	NN	O	O
14	NN	O	O
)	NN	O	O
translocations	NN	O	O
whereas	NN	O	O
the	NN	O	O
MTCP1/A1	NN	O	B-RNA
transcript	NN	O	I-RNA
is	NN	O	O
much	NN	O	O
more	NN	O	O
widely	NN	O	O
expressed	NN	O	O
in	NN	O	O
both	NN	O	O
tumour	NN	O	B-cell_type
and	NN	O	I-cell_type
non	NN	O	I-cell_type
tumour	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
A-T	NN	O	O
and	NN	O	O
non-A-T	NN	O	O
individuals	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Correlation	NN	O	O
of	NN	O	O
lymphocytic	NN	O	O
infiltration	NN	O	O
of	NN	O	O
tumor	NN	O	O
tissue	NN	O	O
with	NN	O	O
the	NN	O	O
hormonal	NN	O	O
and	NN	O	O
metabolic	NN	O	O
state	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
]	NN	O	O

Lymphocyte	NN	O	O
infiltration	NN	O	O
of	NN	O	O
tumor	NN	O	O
was	NN	O	O
studied	NN	O	O
vis-a-vis	NN	O	O
hormone	NN	O	O
metabolic	NN	O	O
status	NN	O	O
,	NN	O	O
tumor	NN	O	O
tissue	NN	O	O
hormone	NN	O	O
sensitivity	NN	O	O
and	NN	O	O
tobacco	NN	O	O
smoking	NN	O	O
,	NN	O	O
in	NN	O	O
113	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
,	NN	O	O
aged	NN	O	O
25-77	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
average	NN	O	O
,	NN	O	O
no	NN	O	O
correlation	NN	O	O
was	NN	O	O
established	NN	O	O
between	NN	O	O
degree	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
infiltration	NN	O	O
in	NN	O	O
breast	NN	O	O
tumor	NN	O	O
and	NN	O	O
age	NN	O	O
and	NN	O	O
menopause	NN	O	O
onset	NN	O	O
.	NN	O	O

In	NN	O	O
smoking	NN	O	O
menopausal	NN	O	O
patients	NN	O	O
,	NN	O	O
lymphocyte	NN	O	O
infiltration	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
higher	NN	O	O
than	NN	O	O
in	NN	O	O
non-smokers	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
direct	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
infiltration	NN	O	O
and	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
progesterone	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
tumor	NN	O	O
.	NN	O	O

Some	NN	O	O
subgroups	NN	O	O
displayed	NN	O	O
a	NN	O	O
direct	NN	O	O
correlation	NN	O	O
between	NN	O	O
infiltration	NN	O	O
and	NN	O	O
sex-binding	NN	O	B-protein
globulin	NN	O	I-protein
,	NN	O	O
cholesterol	NN	O	O
,	NN	O	O
luteinizing	NN	O	O
hormone	NN	O	O
in	NN	O	O
blood	NN	O	O
,	NN	O	O
and	NN	O	O
lean	NN	O	O
body	NN	O	O
mass	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
matched	NN	O	O
by	NN	O	O
an	NN	O	O
inverse	NN	O	O
correlation	NN	O	O
between	NN	O	O
lymphocyte	NN	O	O
infiltration	NN	O	O
and	NN	O	O
blood-thyroid	NN	O	O
hormone	NN	O	O
concentration	NN	O	O
,	NN	O	O
urine	NN	O	O
catecholamines	NN	O	O
and	NN	O	O
free	NN	O	O
cortisol	NN	O	O
excretion	NN	O	O
and	NN	O	O
fat/lean	NN	O	O
body	NN	O	O
mass	NN	O	O
ratio	NN	O	O
.	NN	O	O

Considering	NN	O	O
the	NN	O	O
abovesaid	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
lymphocyte	NN	O	O
ability	NN	O	O
to	NN	O	O
perform	NN	O	O
the	NN	O	O
dual	NN	O	O
function	NN	O	O
of	NN	O	O
immunocytes	NN	O	B-cell_type
and	NN	O	O
hormonocytes	NN	O	B-cell_type
,	NN	O	O
it	NN	O	O
is	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
results	NN	O	O
may	NN	O	O
be	NN	O	O
used	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
infiltration	NN	O	O
and	NN	O	O
research	NN	O	O
in	NN	O	O
means	NN	O	O
of	NN	O	O
its	NN	O	O
control	NN	O	O
.	NN	O	O

-DOCSTART-	O

Gene	NN	O	O
transcription	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
G-protein-coupled	NN	O	B-protein
chemoattractant	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Receptors	NN	O	O
for	NN	O	O
leukocyte	NN	O	B-protein
chemoattractants	NN	O	I-protein
,	NN	O	O
including	NN	O	O
chemokines	NN	O	B-protein
,	NN	O	O
are	NN	O	O
traditionally	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
special	NN	O	O
leukocyte	NN	O	O
functions	NN	O	O
such	NN	O	O
as	NN	O	O
chemotaxis	NN	O	O
,	NN	O	O
degranulation	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
superoxide	NN	O	O
anions	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
these	NN	O	O
G-protein-coupled	NN	O	B-protein
serpentine	NN	O	I-protein
receptors	NN	O	I-protein
have	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
transduce	NN	O	O
signals	NN	O	O
leading	NN	O	O
to	NN	O	O
gene	NN	O	O
transcription	NN	O	O
and	NN	O	O
translation	NN	O	O
in	NN	O	O
leukocytes	NN	O	B-cell_type
.	NN	O	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
are	NN	O	O
activated	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
several	NN	O	O
chemoattractants	NN	O	O
at	NN	O	O
physiologically	NN	O	O
relevant	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
transcription	NN	O	O
factors	NN	O	O
through	NN	O	O
these	NN	O	O
receptors	NN	O	O
involves	NN	O	O
G-protein	NN	O	B-protein
coupling	NN	O	O
and	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

The	NN	O	O
underlying	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
different	NN	O	O
from	NN	O	O
those	NN	O	O
utilized	NN	O	O
by	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
a	NN	O	O
better	NN	O	O
characterized	NN	O	O
cytokine	NN	O	O
that	NN	O	O
induces	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
immediate-early	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Chemoattractants	NN	O	O
stimulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
and	NN	O	O
chemokines	NN	O	B-protein
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
may	NN	O	O
activate	NN	O	O
their	NN	O	O
respective	NN	O	O
receptors	NN	O	O
and	NN	O	O
initiate	NN	O	O
an	NN	O	O
autocrine	NN	O	O
regulatory	NN	O	O
mechanism	NN	O	O
for	NN	O	O
persistent	NN	O	O
cytokine	NN	O	B-protein
and	NN	O	O
chemokine	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Affinity-driven	NN	O	O
peptide	NN	O	O
selection	NN	O	O
of	NN	O	O
an	NN	O	O
NFAT	NN	O	O
inhibitor	NN	O	O
more	NN	O	O
selective	NN	O	O
than	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

The	NN	O	O
flow	NN	O	O
of	NN	O	O
information	NN	O	O
from	NN	O	O
calcium-mobilizing	NN	O	B-protein
receptors	NN	O	I-protein
to	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
of	NN	O	I-DNA
activated	NN	O	I-DNA
T	NN	O	I-DNA
cells	NN	O	I-DNA
(	NN	O	I-DNA
NFAT	NN	O	I-DNA
)	NN	O	I-DNA
-dependent	NN	O	I-DNA
genes	NN	O	I-DNA
is	NN	O	O
critically	NN	O	O
dependent	NN	O	O
on	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
phosphatase	NN	O	B-protein
calcineurin	NN	O	B-protein
and	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NFAT	NN	O	B-protein
.	NN	O	O

A	NN	O	O
high-affinity	NN	O	O
calcineurin	NN	O	B-protein
-binding	NN	O	O
peptide	NN	O	O
was	NN	O	O
selected	NN	O	O
from	NN	O	O
combinatorial	NN	O	O
peptide	NN	O	O
libraries	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
docking	NN	O	I-protein
motif	NN	O	I-protein
of	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

This	NN	O	O
peptide	NN	O	O
potently	NN	O	O
inhibited	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
NFAT	NN	O	B-protein
-dependent	NN	O	O
expression	NN	O	O
of	NN	O	O
endogenous	NN	O	B-DNA
cytokine	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
without	NN	O	O
affecting	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
require	NN	O	O
calcineurin	NN	O	B-protein
but	NN	O	O
not	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

Substitution	NN	O	O
of	NN	O	O
the	NN	O	O
optimized	NN	O	O
peptide	NN	O	O
sequence	NN	O	O
into	NN	O	O
the	NN	O	O
natural	NN	O	O
calcineurin	NN	O	B-protein
docking	NN	O	I-protein
site	NN	O	I-protein
increased	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
responsiveness	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

Compounds	NN	O	O
that	NN	O	O
interfere	NN	O	O
selectively	NN	O	O
with	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
-NFAT	NN	O	B-protein
interaction	NN	O	O
without	NN	O	O
affecting	NN	O	O
calcineurin	NN	O	B-protein
phosphatase	NN	O	B-protein
activity	NN	O	O
may	NN	O	O
be	NN	O	O
useful	NN	O	O
as	NN	O	O
therapeutic	NN	O	O
agents	NN	O	O
that	NN	O	O
are	NN	O	O
less	NN	O	O
toxic	NN	O	O
than	NN	O	O
current	NN	O	O
drugs	NN	O	O
.	NN	O	O

-DOCSTART-	O

9-cis	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
induces	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
secretion	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
THP-1	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Monocyte	NN	O	O
migration	NN	O	O
and	NN	O	O
activation	NN	O	O
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
MCP-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Prior	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
modulated	NN	O	O
by	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
ligands	NN	O	O
that	NN	O	O
act	NN	O	O
through	NN	O	O
extracellular	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
current	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
9-cis	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
ligand	NN	O	O
for	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
retinoid	NN	O	I-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RXR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
markedly	NN	O	O
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
human	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
cultured	NN	O	O
with	NN	O	O
RA	NN	O	O
(	NN	O	O
0.05	NN	O	O
to	NN	O	O
500	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
,	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
induced	NN	O	O
rapidly	NN	O	O
,	NN	O	O
significantly	NN	O	O
,	NN	O	O
and	NN	O	O
dose-dependently	NN	O	O
by	NN	O	O
as	NN	O	O
much	NN	O	O
as	NN	O	O
165-fold	NN	O	O
.	NN	O	O

MCP-1	NN	O	B-RNA
RNA	NN	O	I-RNA
level	NN	O	O
was	NN	O	O
also	NN	O	O
increased	NN	O	O
in	NN	O	O
RA-treated	NN	O	O
cells	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
PPARgamma	NN	O	B-protein
,	NN	O	O
a	NN	O	O
heterodimer	NN	O	O
partner	NN	O	O
of	NN	O	O
RXR	NN	O	B-protein
,	NN	O	O
is	NN	O	O
also	NN	O	O
markedly	NN	O	O
induced	NN	O	O
by	NN	O	O
RA	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
BRL49653	NN	O	O
,	NN	O	O
a	NN	O	O
PPARgamma	NN	O	B-protein
ligand	NN	O	O
,	NN	O	O
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
MCP-1	NN	O	B-protein
secretion	NN	O	O
either	NN	O	O
alone	NN	O	O
or	NN	O	O
to	NN	O	O
modify	NN	O	O
the	NN	O	O
expression	NN	O	O
level	NN	O	O
induced	NN	O	O
by	NN	O	O
RA	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
BRL49653	NN	O	O
significantly	NN	O	O
increased	NN	O	O
MCP-1	NN	O	B-protein
(	NN	O	O
biotinylated	NN	O	B-protein
MCP-1	NN	O	I-protein
)	NN	O	O
binding	NN	O	O
to	NN	O	O
THP-1	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
RA	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

Other	NN	O	O
peroxisome	NN	O	B-protein
proliferator	NN	O	I-protein
activated	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
PPAR	NN	O	B-protein
)	NN	O	O
ligands	NN	O	O
,	NN	O	O
15d-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
and	NN	O	O
troglitazone	NN	O	O
(	NN	O	O
PPARgamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
Wy14	NN	O	O
,	NN	O	O
643	NN	O	O
(	NN	O	O
PPARalpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
PD195599	NN	O	O
(	NN	O	O
PPARbeta	NN	O	B-protein
)	NN	O	O
inhibited	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
by	NN	O	O
RA	NN	O	O
.	NN	O	O

RA	NN	O	O
's	NN	O	O
effect	NN	O	O
on	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	O
elutriated	NN	O	O
monocytes	NN	O	O
were	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
studies	NN	O	O
identify	NN	O	O
RA	NN	O	O
as	NN	O	O
a	NN	O	O
nuclear	NN	O	O
signal	NN	O	O
for	NN	O	O
MCP-1	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
undifferentiated	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
studies	NN	O	O
also	NN	O	O
suggest	NN	O	O
monocyte	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
induced	NN	O	O
through	NN	O	O
RA	NN	O	O
may	NN	O	O
modulate	NN	O	O
cell	NN	O	O
migration	NN	O	O
.	NN	O	O

-DOCSTART-	O

Classification	NN	O	O
of	NN	O	O
IVS1-10T	NN	O	O
--	NN	O	O
>	NN	O	O
C	NN	O	O
as	NN	O	O
a	NN	O	O
polymorphism	NN	O	O
of	NN	O	O
BRCA1	NN	O	B-DNA
.	NN	O	O

Mutations	NN	O	O
inactivating	NN	O	O
the	NN	O	O
tumor	NN	O	B-DNA
suppressor	NN	O	I-DNA
gene	NN	O	I-DNA
BRCA1	NN	O	B-DNA
may	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
disease	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
80	NN	O	O
%	NN	O	O
of	NN	O	O
familial	NN	O	O
ovarian	NN	O	O
cancer	NN	O	O
cases	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
syndrome	NN	O	O
,	NN	O	O
tumorigenesis	NN	O	O
classically	NN	O	O
initiates	NN	O	O
from	NN	O	O
an	NN	O	O
inherited	NN	O	O
mutation	NN	O	O
in	NN	O	O
one	NN	O	O
allele	NN	O	B-DNA
followed	NN	O	O
by	NN	O	O
somatic	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	B-DNA
allele	NN	O	I-DNA
.	NN	O	O

Sequencing	NN	O	O
of	NN	O	O
BRCA1	NN	O	B-DNA
amplified	NN	O	O
from	NN	O	O
genomic	NN	O	B-DNA
DNA	NN	O	I-DNA
of	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
microdissected	NN	O	B-cell_type
ovarian	NN	O	I-cell_type
tumor	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
a	NN	O	O
familial	NN	O	O
ovarian	NN	O	O
cancer	NN	O	O
patient	NN	O	O
revealed	NN	O	O
three	NN	O	O
,	NN	O	O
rare	NN	O	O
heterozygous	NN	O	O
DNA	NN	O	O
variations	NN	O	O
(	NN	O	O
2418delA	NN	O	O
,	NN	O	O
233G	NN	O	O
--	NN	O	O
>	NN	O	O
A	NN	O	O
,	NN	O	O
and	NN	O	O
IVS1-10T	NN	O	O
--	NN	O	O
>	NN	O	O
C	NN	O	O
)	NN	O	O
in	NN	O	O
both	NN	O	O
tumor	NN	O	B-DNA
and	NN	O	I-DNA
constitutional	NN	O	I-DNA
(	NN	O	I-DNA
lymphocyte	NN	O	I-DNA
)	NN	O	I-DNA
DNA	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
both	NN	O	O
copies	NN	O	O
of	NN	O	O
BRCA1	NN	O	B-DNA
were	NN	O	O
retained	NN	O	O
in	NN	O	O
tumor	NN	O	O
.	NN	O	O

Haplotype	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
and	NN	O	O
four	NN	O	O
siblings	NN	O	O
assigned	NN	O	O
2418delA	NN	O	O
to	NN	O	O
one	NN	O	O
copy	NN	O	O
of	NN	O	O
BRCA1	NN	O	B-DNA
and	NN	O	O
233G	NN	O	O
--	NN	O	O
>	NN	O	O
A	NN	O	O
and	NN	O	O
IVS1-10T	NN	O	O
--	NN	O	O
>	NN	O	O
C	NN	O	O
to	NN	O	O
the	NN	O	O
other	NN	O	O
.	NN	O	O

The	NN	O	O
DNA	NN	O	O
change	NN	O	O
,	NN	O	O
2418delA	NN	O	O
,	NN	O	O
is	NN	O	O
considered	NN	O	O
a	NN	O	O
mutation	NN	O	O
that	NN	O	O
inactivated	NN	O	O
one	NN	O	O
BRCA1	NN	O	B-DNA
allele	NN	O	O
because	NN	O	O
it	NN	O	O
caused	NN	O	O
a	NN	O	O
frameshift	NN	O	O
and	NN	O	O
generation	NN	O	O
of	NN	O	O
a	NN	O	O
premature	NN	O	B-DNA
stop	NN	O	I-DNA
codon	NN	O	I-DNA
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
truncated	NN	O	O
peptide	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
protein	NN	O	O
truncation	NN	O	O
test	NN	O	O
.	NN	O	O

The	NN	O	O
DNA	NN	O	O
variation	NN	O	O
,	NN	O	O
233G	NN	O	O
--	NN	O	O
>	NN	O	O
A	NN	O	O
,	NN	O	O
does	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
an	NN	O	O
amino	NN	O	O
acid	NN	O	O
change	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
considered	NN	O	O
a	NN	O	O
benign	NN	O	O
polymorphism	NN	O	O
.	NN	O	O

IVS1-10T	NN	O	O
--	NN	O	O
>	NN	O	O
C	NN	O	O
is	NN	O	O
a	NN	O	O
unique	NN	O	O
BRCA1	NN	O	B-DNA
change	NN	O	O
that	NN	O	O
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
last	NN	O	O
nucleotide	NN	O	O
of	NN	O	O
a	NN	O	O
consensus	NN	O	B-DNA
sequence	NN	O	I-DNA
for	NN	O	O
a	NN	O	O
branch	NN	O	B-DNA
site	NN	O	I-DNA
critical	NN	O	O
for	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
IVS1-10T	NN	O	O
--	NN	O	O
>	NN	O	O
C	NN	O	O
deleteriously	NN	O	O
affected	NN	O	O
BRCA1	NN	O	B-DNA
splicing	NN	O	O
or	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
thereby	NN	O	O
inactivated	NN	O	O
the	NN	O	O
other	NN	O	O
BRCA1	NN	O	B-DNA
allele	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
the	NN	O	O
technique	NN	O	O
of	NN	O	O
reverse	NN	O	O
transcription-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
with	NN	O	O
RNA	NN	O	O
isolated	NN	O	O
from	NN	O	O
lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
patient	NN	O	O
and	NN	O	O
of	NN	O	O
controls	NN	O	O
,	NN	O	O
no	NN	O	O
evidence	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
IVS1-10TC	NN	O	O
abnormally	NN	O	O
disrupted	NN	O	O
mRNA	NN	O	B-RNA
splicing	NN	O	O
or	NN	O	O
caused	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
BRCA1	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
IVS1-10T	NN	O	O
--	NN	O	O
>	NN	O	O
C	NN	O	O
is	NN	O	O
not	NN	O	O
harmful	NN	O	O
to	NN	O	O
BRCA1	NN	O	B-DNA
function	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
classified	NN	O	O
a	NN	O	O
benign	NN	O	O
polymorphism	NN	O	O
.	NN	O	O

Retention	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
BRCA1	NN	O	B-DNA
allele	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
tumor	NN	O	O
with	NN	O	O
the	NN	O	O
heterozygous	NN	O	O
germline	NN	O	O
BRCA1	NN	O	B-DNA
mutation	NN	O	O
,	NN	O	O
2418delA	NN	O	O
,	NN	O	O
indicated	NN	O	O
that	NN	O	O
mutational	NN	O	O
inactivation	NN	O	O
of	NN	O	O
both	NN	O	O
BRCA1	NN	O	B-DNA
alleles	NN	O	I-DNA
was	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
tumorigenesis	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
possible	NN	O	O
that	NN	O	O
the	NN	O	O
normal	NN	O	B-DNA
allele	NN	O	I-DNA
may	NN	O	O
be	NN	O	O
functionally	NN	O	O
inactivated	NN	O	O
by	NN	O	O
a	NN	O	O
nonmutational	NN	O	O
mechanism	NN	O	O
.	NN	O	O

-DOCSTART-	O

Myb-transformed	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
into	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

Immune	NN	O	O
induction	NN	O	O
is	NN	O	O
effected	NN	O	O
through	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
specific	NN	O	O
receptors	NN	O	B-protein
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
thymus-derived	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Cells	NN	O	O
most	NN	O	O
able	NN	O	O
to	NN	O	O
ingest	NN	O	O
,	NN	O	O
process	NN	O	O
,	NN	O	O
and	NN	O	O
present	NN	O	O
antigen	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
mononuclear	NN	O	B-cell_type
phagocyte/neutrophil	NN	O	I-cell_type
series	NN	O	I-cell_type
.	NN	O	O

For	NN	O	O
example	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
DC	NN	O	B-cell_type
)	NN	O	O
can	NN	O	O
be	NN	O	O
found	NN	O	O
in	NN	O	O
colonies	NN	O	O
of	NN	O	O
GM-CSF-responsive	NN	O	B-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
under	NN	O	O
experimental	NN	O	O
conditions	NN	O	O
are	NN	O	O
routinely	NN	O	O
expanded	NN	O	O
as	NN	O	O
a	NN	O	O
population	NN	O	O
in	NN	O	O
vitro	NN	O	O
from	NN	O	O
GM-CSF-responsive	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
address	NN	O	O
the	NN	O	O
question	NN	O	O
of	NN	O	O
DC	NN	O	B-cell_type
lineage	NN	O	I-cell_type
and	NN	O	O
to	NN	O	O
determine	NN	O	O
what	NN	O	O
genes	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
generated	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
GM-CSF-responsive	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
in	NN	O	O
a	NN	O	O
homogeneous	NN	O	O
manner	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
cloned	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
are	NN	O	O
derived	NN	O	O
from	NN	O	O
12-day	NN	O	O
fetal	NN	O	O
liver	NN	O	O
and	NN	O	O
are	NN	O	O
transformed	NN	O	O
with	NN	O	O
a	NN	O	O
truncated	NN	O	O
form	NN	O	O
of	NN	O	O
c-myb	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
lacks	NN	O	O
the	NN	O	O
normal	NN	O	O
autoregulatory	NN	O	B-DNA
sequences	NN	O	I-DNA
.	NN	O	O

As	NN	O	O
far	NN	O	O
as	NN	O	O
we	NN	O	O
know	NN	O	O
,	NN	O	O
these	NN	O	O
myb-transformed	NN	O	B-cell_line
hemopoi-etic	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
MTHC	NN	O	B-cell_line
)	NN	O	O
differ	NN	O	O
from	NN	O	O
normal	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
unregulated	NN	O	O
expression	NN	O	O
of	NN	O	O
myb	NN	O	O
,	NN	O	O
a	NN	O	O
gene	NN	O	O
whose	NN	O	O
expression	NN	O	O
is	NN	O	O
obligatory	NN	O	O
for	NN	O	O
proliferation	NN	O	O
of	NN	O	O
hemopoietic	NN	O	O
cells	NN	O	O
.	NN	O	O

MTHC	NN	O	B-cell_line
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
will	NN	O	O
differentiate	NN	O	O
into	NN	O	O
cells	NN	O	O
that	NN	O	O
have	NN	O	O
many	NN	O	O
of	NN	O	O
the	NN	O	O
properties	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

When	NN	O	O
the	NN	O	O
same	NN	O	O
MTHC	NN	O	B-cell_line
lines	NN	O	I-cell_line
are	NN	O	O
exposed	NN	O	O
to	NN	O	O
TNF-alpha	NN	O	B-protein
in	NN	O	O
combination	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
the	NN	O	O
cells	NN	O	O
instead	NN	O	O
become	NN	O	O
DC	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
differentiated	NN	O	O
DC	NN	O	B-cell_type
are	NN	O	O
potent	NN	O	O
presenters	NN	O	O
of	NN	O	O
antigen	NN	O	O
in	NN	O	O
mixed	NN	O	O
lymphocyte	NN	O	O
reactions	NN	O	O
and	NN	O	O
of	NN	O	O
soluble	NN	O	O
antigen	NN	O	O
to	NN	O	O
specific	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
cells	NN	O	O
with	NN	O	O
the	NN	O	O
properties	NN	O	O
of	NN	O	O
both	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
DC	NN	O	B-cell_type
can	NN	O	O
be	NN	O	O
derived	NN	O	O
from	NN	O	O
a	NN	O	O
single	NN	O	O
type	NN	O	O
of	NN	O	O
GM-CSF-responsive	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
this	NN	O	O
MTHC	NN	O	B-cell_line
system	NN	O	I-cell_line
to	NN	O	O
analyze	NN	O	O
differences	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
as	NN	O	O
the	NN	O	O
cells	NN	O	O
mature	NN	O	O
along	NN	O	O
the	NN	O	O
DC	NN	O	B-cell_type
and	NN	O	O
macrophage	NN	O	B-cell_type
pathways	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
distinctive	NN	O	O
pattern	NN	O	O
of	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
cDNAs	NN	O	B-DNA
is	NN	O	O
evident	NN	O	O
where	NN	O	O
macrophage-specific	NN	O	B-DNA
cDNAs	NN	O	I-DNA
are	NN	O	O
homologous	NN	O	O
to	NN	O	O
genes	NN	O	O
encoding	NN	O	O
cytoskeletal	NN	O	B-protein
and	NN	O	I-protein
cell-surface	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
DC	NN	O	B-cell_type
-specific	NN	O	O
cDNAs	NN	O	B-DNA
are	NN	O	O
homologous	NN	O	O
to	NN	O	O
signaling	NN	O	B-DNA
,	NN	O	I-DNA
chemokine	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
IFN-gamma-inducible	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
discuss	NN	O	O
the	NN	O	O
utility	NN	O	O
of	NN	O	O
MTHC	NN	O	B-cell_line
in	NN	O	O
analyzing	NN	O	O
the	NN	O	O
relationships	NN	O	O
between	NN	O	O
DC	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
DC	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
represent	NN	O	O
extreme	NN	O	O
phenotypes	NN	O	O
in	NN	O	O
a	NN	O	O
spectrum	NN	O	O
of	NN	O	O
antigen	NN	O	B-cell_type
handling	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
are	NN	O	O
somewhat	NN	O	O
interchangeable	NN	O	O
,	NN	O	O
depending	NN	O	O
on	NN	O	O
their	NN	O	O
immediate	NN	O	O
environment	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Legionella	NN	O	O
pneumophila	NN	O	O
rpoS	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
required	NN	O	O
for	NN	O	O
growth	NN	O	O
within	NN	O	O
Acanthamoeba	NN	O	O
castellanii	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
regulatory	NN	O	O
networks	NN	O	O
in	NN	O	O
Legionella	NN	O	O
pneumophila	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
homolog	NN	O	O
of	NN	O	O
the	NN	O	O
Escherichia	NN	O	B-protein
coli	NN	O	I-protein
stress	NN	O	I-protein
and	NN	O	I-protein
stationary-phase	NN	O	I-protein
sigma	NN	O	I-protein
factor	NN	O	I-protein
RpoS	NN	O	B-protein
was	NN	O	O
identified	NN	O	O
by	NN	O	O
complementation	NN	O	O
of	NN	O	O
an	NN	O	O
E.	NN	O	O
coli	NN	O	O
rpoS	NN	O	B-DNA
mutation	NN	O	O
.	NN	O	O

An	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
approximately	NN	O	O
60	NN	O	O
%	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
E.	NN	O	B-DNA
coli	NN	O	I-DNA
rpoS	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
identified	NN	O	O
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
L.	NN	O	O
pneumophila	NN	O	O
RpoS	NN	O	B-protein
increased	NN	O	O
in	NN	O	O
stationary	NN	O	O
phase	NN	O	O
.	NN	O	O

An	NN	O	O
insertion	NN	O	O
mutation	NN	O	O
was	NN	O	O
constructed	NN	O	O
in	NN	O	O
the	NN	O	O
rpoS	NN	O	B-DNA
gene	NN	O	I-DNA
on	NN	O	O
the	NN	O	O
chromosome	NN	O	O
of	NN	O	O
L.	NN	O	O
pneumophila	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
this	NN	O	O
mutant	NN	O	O
strain	NN	O	O
to	NN	O	O
survive	NN	O	O
various	NN	O	O
stress	NN	O	O
conditions	NN	O	O
was	NN	O	O
assayed	NN	O	O
and	NN	O	O
compared	NN	O	O
with	NN	O	O
results	NN	O	O
for	NN	O	O
the	NN	O	O
wild-type	NN	O	O
strain	NN	O	O
.	NN	O	O

Both	NN	O	O
the	NN	O	O
mutant	NN	O	O
and	NN	O	O
wild-type	NN	O	O
strains	NN	O	O
were	NN	O	O
more	NN	O	O
resistant	NN	O	O
to	NN	O	O
stress	NN	O	O
when	NN	O	O
in	NN	O	O
stationary	NN	O	O
phase	NN	O	O
than	NN	O	O
when	NN	O	O
in	NN	O	O
the	NN	O	O
logarithmic	NN	O	O
phase	NN	O	O
of	NN	O	O
growth	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
indicates	NN	O	O
that	NN	O	O
L.	NN	O	O
pneumophila	NN	O	O
RpoS	NN	O	B-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
a	NN	O	O
stationary-phase-dependent	NN	O	O
resistance	NN	O	O
to	NN	O	O
stress	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
mutant	NN	O	O
strain	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
kill	NN	O	O
HL-60-	NN	O	B-cell_type
and	NN	O	I-cell_type
THP-1-derived	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
it	NN	O	O
could	NN	O	O
not	NN	O	O
replicate	NN	O	O
within	NN	O	O
a	NN	O	O
protozoan	NN	O	O
host	NN	O	O
,	NN	O	O
Acanthamoeba	NN	O	O
castellanii	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
L.	NN	O	O
pneumophila	NN	O	O
possesses	NN	O	O
a	NN	O	O
growth	NN	O	O
phase-dependent	NN	O	O
resistance	NN	O	O
to	NN	O	O
stress	NN	O	O
that	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
RpoS	NN	O	B-protein
control	NN	O	O
and	NN	O	O
that	NN	O	O
RpoS	NN	O	B-protein
likely	NN	O	O
regulates	NN	O	O
genes	NN	O	O
that	NN	O	O
enable	NN	O	O
it	NN	O	O
to	NN	O	O
survive	NN	O	O
in	NN	O	O
the	NN	O	O
environment	NN	O	O
within	NN	O	O
protozoa	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
rpoS	NN	O	B-DNA
in	NN	O	O
L.	NN	O	O
pneumophila	NN	O	O
is	NN	O	O
very	NN	O	O
different	NN	O	O
from	NN	O	O
what	NN	O	O
has	NN	O	O
previously	NN	O	O
been	NN	O	O
reported	NN	O	O
for	NN	O	O
E.	NN	O	O
coli	NN	O	O
rpoS	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Monoallelic	NN	O	O
expression	NN	O	O
of	NN	O	O
Pax5	NN	O	B-protein
:	NN	O	O
a	NN	O	O
paradigm	NN	O	O
for	NN	O	O
the	NN	O	O
haploinsufficiency	NN	O	O
of	NN	O	O
mammalian	NN	O	B-DNA
Pax	NN	O	I-DNA
genes	NN	O	I-DNA
?	NN	O	O

It	NN	O	O
is	NN	O	O
generally	NN	O	O
assumed	NN	O	O
that	NN	O	O
most	NN	O	O
mammalian	NN	O	B-DNA
genes	NN	O	I-DNA
are	NN	O	O
transcribed	NN	O	O
from	NN	O	O
both	NN	O	O
alleles	NN	O	B-DNA
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
the	NN	O	O
diploid	NN	O	O
state	NN	O	O
of	NN	O	O
the	NN	O	O
genome	NN	O	O
offers	NN	O	O
the	NN	O	O
advantage	NN	O	O
that	NN	O	O
a	NN	O	O
loss-of-function	NN	O	O
mutation	NN	O	O
in	NN	O	O
one	NN	O	O
allele	NN	O	B-DNA
can	NN	O	O
be	NN	O	O
compensated	NN	O	O
for	NN	O	O
by	NN	O	O
the	NN	O	O
remaining	NN	O	O
wild-type	NN	O	B-DNA
allele	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
same	NN	O	O
gene	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
the	NN	O	O
vast	NN	O	O
majority	NN	O	O
of	NN	O	O
human	NN	O	O
disease	NN	O	O
syndromes	NN	O	O
and	NN	O	O
engineered	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
genome	NN	O	I-DNA
are	NN	O	O
recessive	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
recessiveness	NN	O	O
is	NN	O	O
the	NN	O	O
'default	NN	O	O
'	NN	O	O
state	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
minority	NN	O	O
of	NN	O	O
genes	NN	O	O
are	NN	O	O
semi-dominant	NN	O	O
,	NN	O	O
as	NN	O	O
heterozygous	NN	O	O
loss-of-function	NN	O	O
mutation	NN	O	O
in	NN	O	O
these	NN	O	O
genes	NN	O	O
leads	NN	O	O
to	NN	O	O
phenotypic	NN	O	O
abnormalities	NN	O	O
.	NN	O	O

This	NN	O	O
condition	NN	O	O
,	NN	O	O
known	NN	O	O
as	NN	O	O
haploinsufficiency	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
described	NN	O	O
for	NN	O	O
five	NN	O	O
of	NN	O	O
the	NN	O	O
nine	NN	O	O
mammalian	NN	O	B-DNA
Pax	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
mouse	NN	O	O
developmental	NN	O	O
mutants	NN	O	O
and	NN	O	O
human	NN	O	O
disease	NN	O	O
syndromes	NN	O	O
.	NN	O	O

Recently	NN	O	O
we	NN	O	O
have	NN	O	O
reported	NN	O	O
that	NN	O	O
the	NN	O	O
Pax5	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
subject	NN	O	O
to	NN	O	O
allele	NN	O	B-DNA
-specific	NN	O	O
regulation	NN	O	O
during	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

Pax5	NN	O	B-protein
is	NN	O	O
predominantly	NN	O	O
transcribed	NN	O	O
from	NN	O	O
only	NN	O	O
one	NN	O	O
of	NN	O	O
its	NN	O	O
two	NN	O	O
alleles	NN	O	B-DNA
in	NN	O	O
early	NN	O	B-cell_type
B-lymphoid	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
and	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
it	NN	O	O
transiently	NN	O	O
switches	NN	O	O
to	NN	O	O
a	NN	O	O
biallelic	NN	O	O
mode	NN	O	O
of	NN	O	O
transcription	NN	O	O
in	NN	O	O
pre-B	NN	O	B-cell_type
and	NN	O	I-cell_type
immature	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

As	NN	O	O
a	NN	O	O
consequence	NN	O	O
,	NN	O	O
B-lymphoid	NN	O	O
tissues	NN	O	O
are	NN	O	O
mosaic	NN	O	O
with	NN	O	O
regard	NN	O	O
to	NN	O	O
the	NN	O	O
transcribed	NN	O	O
allele	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
heterozygous	NN	O	O
mutation	NN	O	O
of	NN	O	O
Pax5	NN	O	B-protein
therefore	NN	O	O
results	NN	O	O
in	NN	O	O
deletion	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
expressing	NN	O	O
only	NN	O	O
the	NN	O	O
mutant	NN	O	B-DNA
allele	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
allele	NN	O	B-DNA
-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
Pax5	NN	O	B-protein
raises	NN	O	O
the	NN	O	O
intriguing	NN	O	O
possibility	NN	O	O
that	NN	O	O
monoallelic	NN	O	O
expression	NN	O	O
may	NN	O	O
also	NN	O	O
be	NN	O	O
the	NN	O	O
mechanism	NN	O	O
causing	NN	O	O
the	NN	O	O
haploinsufficiency	NN	O	O
of	NN	O	O
other	NN	O	O
Pax	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
,	NN	O	O
we	NN	O	O
discuss	NN	O	O
different	NN	O	O
models	NN	O	O
accounting	NN	O	O
for	NN	O	O
the	NN	O	O
haploinsufficiency	NN	O	O
of	NN	O	O
mammalian	NN	O	B-DNA
Pax	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
provide	NN	O	O
further	NN	O	O
evidence	NN	O	O
in	NN	O	O
support	NN	O	O
of	NN	O	O
the	NN	O	O
allele	NN	O	B-DNA
-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
Pax5	NN	O	B-protein
and	NN	O	O
discuss	NN	O	O
the	NN	O	O
implication	NN	O	O
of	NN	O	O
these	NN	O	O
findings	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
recent	NN	O	O
literature	NN	O	O
describing	NN	O	O
the	NN	O	O
stochastic	NN	O	O
and	NN	O	O
monoallelic	NN	O	O
activation	NN	O	O
of	NN	O	O
other	NN	O	O
hematopoietic	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
by	NN	O	O
IL-4	NN	O	B-protein
relies	NN	O	O
on	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
function	NN	O	O
but	NN	O	O
not	NN	O	O
on	NN	O	O
receptor	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
and	NN	O	O
can	NN	O	O
contribute	NN	O	O
to	NN	O	O
both	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
gene	NN	O	O
regulation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
mechanisms	NN	O	O
and	NN	O	O
consequences	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
activation	NN	O	O
through	NN	O	O
the	NN	O	O
human	NN	O	B-protein
IL-4	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-4R	NN	O	B-protein
)	NN	O	O
.	NN	O	O

By	NN	O	O
functionally	NN	O	O
expressing	NN	O	O
receptor	NN	O	B-protein
mutants	NN	O	I-protein
in	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
pro-B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Ba/F3	NN	O	B-cell_line
,	NN	O	O
we	NN	O	O
could	NN	O	O
show	NN	O	O
that	NN	O	O
phosphorylated	NN	O	O
tyrosine	NN	O	O
residues	NN	O	O
within	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
are	NN	O	O
dispensable	NN	O	O
for	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
STAT5	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
disruption	NN	O	O
of	NN	O	O
a	NN	O	O
membrane-proximal	NN	O	B-protein
proline-rich	NN	O	I-protein
sequence	NN	O	I-protein
motif	NN	O	I-protein
(	NN	O	O
'	NN	O	O
box1	NN	O	B-protein
'	NN	O	O
)	NN	O	O
in	NN	O	O
either	NN	O	O
subunit	NN	O	O
of	NN	O	O
the	NN	O	O
bipartite	NN	O	B-protein
IL-4R	NN	O	I-protein
abolished	NN	O	O
not	NN	O	O
only	NN	O	O
ligand-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
kinases	NN	O	I-protein
JAK1	NN	O	B-protein
and	NN	O	O
JAK3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
also	NN	O	O
IL-4	NN	O	B-protein
-triggered	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
and	NN	O	O
concomitant	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

A	NN	O	O
dominant-negative	NN	O	B-protein
version	NN	O	I-protein
of	NN	O	O
STAT5b	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
STAT5a	NN	O	B-protein
,	NN	O	O
interfered	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
in	NN	O	O
Ba/F3	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
an	NN	O	O
involvement	NN	O	O
of	NN	O	O
STAT5b	NN	O	B-protein
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
through	NN	O	O
IL-4R	NN	O	B-protein
.	NN	O	O

Reporter	NN	O	O
gene	NN	O	O
experiments	NN	O	O
finally	NN	O	O
showed	NN	O	O
that	NN	O	O
transcription	NN	O	O
from	NN	O	O
promoters	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
target	NN	O	O
genes	NN	O	O
can	NN	O	O
be	NN	O	O
specifically	NN	O	O
induced	NN	O	O
by	NN	O	O
challenging	NN	O	O
cells	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
both	NN	O	O
STAT5a	NN	O	B-protein
and	NN	O	O
STAT5b	NN	O	B-protein
can	NN	O	O
contribute	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
-triggered	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
by	NN	O	O
a	NN	O	O
dominant-negative	NN	O	O
form	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

An	NN	O	O
inhibitory	NN	O	O
,	NN	O	O
``	NN	O	O
dominant-negative	NN	O	O
,	NN	O	O
''	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
catalytic	NN	O	I-protein
(	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
was	NN	O	O
prepared	NN	O	O
,	NN	O	O
which	NN	O	O
lacks	NN	O	O
the	NN	O	O
calmodulin-binding	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
autoinhibitory	NN	O	B-protein
domain	NN	O	I-protein
and	NN	O	O
most	NN	O	O
of	NN	O	O
its	NN	O	O
catalytic	NN	O	O
core	NN	O	O
but	NN	O	O
possesses	NN	O	O
the	NN	O	O
regulatory	NN	O	B-protein
(	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
binding	NN	O	O
domain	NN	O	O
.	NN	O	O

When	NN	O	O
tested	NN	O	O
for	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
block	NN	O	O
calcineurin	NN	O	B-protein
-dependent	NN	O	O
signaling	NN	O	O
in	NN	O	O
Jurkat	NN	O	O
cells	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
``	NN	O	B-DNA
B-subunit	NN	O	I-DNA
knock-out	NN	O	I-DNA
''	NN	O	I-DNA
(	NN	O	I-DNA
BKO	NN	O	I-DNA
)	NN	O	I-DNA
construct	NN	O	I-DNA
suppressed	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
activity	NN	O	O
driven	NN	O	O
by	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
the	NN	O	O
pivotal	NN	O	O
promoter	NN	O	B-DNA
element	NN	O	I-DNA
for	NN	O	O
interleukin	NN	O	B-DNA
(	NN	O	I-DNA
IL	NN	O	I-DNA
)	NN	O	I-DNA
-2	NN	O	I-DNA
gene	NN	O	I-DNA
induction	NN	O	O
.	NN	O	O

Immunoprecipitation	NN	O	O
of	NN	O	O
epitope-labeled	NN	O	B-protein
BKO	NN	O	I-protein
demonstrated	NN	O	O
for	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
tight	NN	O	O
complex	NN	O	O
with	NN	O	O
endogenous	NN	O	B-protein
B	NN	O	I-protein
subunit	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
an	NN	O	O
inhibitory	NN	O	O
mechanism	NN	O	O
that	NN	O	O
involves	NN	O	O
the	NN	O	O
sequestration	NN	O	O
of	NN	O	O
the	NN	O	O
B	NN	O	B-protein
subunit	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
sharply	NN	O	O
reduced	NN	O	O
NF-AT	NN	O	B-protein
activity	NN	O	O
produced	NN	O	O
by	NN	O	O
co-transfecting	NN	O	B-DNA
BKO	NN	O	I-DNA
could	NN	O	O
be	NN	O	O
``	NN	O	O
rescued	NN	O	O
''	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
transfected	NN	O	O
B	NN	O	B-protein
subunit	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
depletion	NN	O	O
of	NN	O	O
this	NN	O	O
subunit	NN	O	O
was	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
inhibition	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
the	NN	O	O
potential	NN	O	O
utility	NN	O	O
of	NN	O	O
agents	NN	O	O
that	NN	O	O
disrupt	NN	O	O
calcineurin	NN	O	B-protein
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
by	NN	O	O
blocking	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
catalytically	NN	O	B-protein
active	NN	O	I-protein
dimer	NN	O	I-protein
of	NN	O	O
calcineurin	NN	O	B-protein
A	NN	O	I-protein
and	NN	O	I-protein
B	NN	O	I-protein
subunits	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Evaluation	NN	O	O
of	NN	O	O
monoclonal	NN	O	O
anti-D	NN	O	O
reagents	NN	O	O
using	NN	O	O
D	NN	O	B-cell_line
variant	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Monoclonal	NN	O	B-protein
anti-D	NN	O	I-protein
antibodies	NN	O	I-protein
submitted	NN	O	O
to	NN	O	O
the	NN	O	O
Third	NN	O	O
Monoclonal	NN	O	O
International	NN	O	O
Workshop	NN	O	O
were	NN	O	O
evaluated	NN	O	O
against	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
D	NN	O	B-cell_line
variant	NN	O	I-cell_line
cells	NN	O	I-cell_line
using	NN	O	O
standard	NN	O	O
serological	NN	O	O
techniques	NN	O	O
.	NN	O	O

The	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
were	NN	O	O
able	NN	O	O
to	NN	O	O
discriminate	NN	O	O
between	NN	O	O
the	NN	O	O
cells	NN	O	O
of	NN	O	O
Categories	NN	O	B-cell_line
Va	NN	O	I-cell_line
,	NN	O	I-cell_line
VI	NN	O	I-cell_line
and	NN	O	I-cell_line
DFR	NN	O	I-cell_line
but	NN	O	I-cell_line
not	NN	O	I-cell_line
Category	NN	O	I-cell_line
III	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Cells	NN	O	O
within	NN	O	O
each	NN	O	O
category	NN	O	O
did	NN	O	O
not	NN	O	O
give	NN	O	O
any	NN	O	O
aberrant	NN	O	O
results	NN	O	O
.	NN	O	O

The	NN	O	O
Rh	NN	O	B-cell_line
:	NN	O	I-cell_line
33	NN	O	I-cell_line
cells	NN	O	I-cell_line
behaved	NN	O	O
as	NN	O	O
normal	NN	O	B-cell_type
Rh	NN	O	I-cell_type
(	NN	O	I-cell_type
D	NN	O	I-cell_type
)	NN	O	I-cell_type
positive	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Interleukin	NN	O	B-protein
10	NN	O	I-protein
induced	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
activation	NN	O	O
of	NN	O	O
divergent	NN	O	B-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

IL-10	NN	O	B-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
mediator	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cell	NN	O	I-cell_type
growth	NN	O	O
and	NN	O	O
plasma	NN	O	O
cell	NN	O	O
formation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
signal	NN	O	O
transduction	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
protooncogene	NN	O	B-DNA
c-fos	NN	O	I-DNA
was	NN	O	O
investigated	NN	O	O
,	NN	O	O
because	NN	O	O
Fos	NN	O	O
plays	NN	O	O
a	NN	O	O
potential	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

B	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
purified	NN	O	O
from	NN	O	O
buffy	NN	O	O
coat	NN	O	O
preparations	NN	O	O
of	NN	O	O
healthy	NN	O	O
blood	NN	O	O
donors	NN	O	O
by	NN	O	O
positive	NN	O	O
selection	NN	O	O
using	NN	O	O
an	NN	O	O
anti	NN	O	B-protein
CD20	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
and	NN	O	O
a	NN	O	O
MiniMACS	NN	O	O
separation	NN	O	O
unit	NN	O	O
.	NN	O	O

B	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
prestimulated	NN	O	O
with	NN	O	O
SAC	NN	O	O
for	NN	O	O
48	NN	O	O
hrs	NN	O	O
.	NN	O	O

Then	NN	O	O
,	NN	O	O
cells	NN	O	O
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
medium	NN	O	O
or	NN	O	O
IL-10	NN	O	B-protein
(	NN	O	O
100	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
for	NN	O	O
10	NN	O	O
to	NN	O	O
120	NN	O	O
min	NN	O	O
.	NN	O	O

RNA	NN	O	B-RNA
was	NN	O	O
extracted	NN	O	O
by	NN	O	O
phenol/chloroform	NN	O	O
and	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
PCR	NN	O	O
assisted	NN	O	O
mRNA	NN	O	O
assay	NN	O	O
.	NN	O	O

A	NN	O	O
significant	NN	O	O
2-4	NN	O	O
fold	NN	O	O
increase	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
was	NN	O	O
observed	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
of	NN	O	O
stimulation	NN	O	O
with	NN	O	O
IL-10	NN	O	B-protein
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

After	NN	O	O
2	NN	O	O
hrs	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
declined	NN	O	O
to	NN	O	O
basal	NN	O	O
levels	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
was	NN	O	O
dose-dependent	NN	O	O
with	NN	O	O
a	NN	O	O
maximum	NN	O	O
stimulation	NN	O	O
using	NN	O	O
100	NN	O	O
ng/ml	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

The	NN	O	O
IL-10	NN	O	B-protein
effect	NN	O	O
on	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
was	NN	O	O
not	NN	O	O
blocked	NN	O	O
by	NN	O	O
polymyxin	NN	O	O
B	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
genistein	NN	O	O
(	NN	O	O
10	NN	O	O
microM	NN	O	O
)	NN	O	O
a	NN	O	O
complete	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
induced	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
H-7	NN	O	O
(	NN	O	O
10	NN	O	O
microM	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
serine/threonine	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
significantly	NN	O	O
blocked	NN	O	O
IL-10	NN	O	B-protein
mediated	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
induces	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B-cells	NN	O	I-cell_type
by	NN	O	O
activation	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
and	NN	O	O
serine/threonine	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

Since	NN	O	O
this	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
report	NN	O	O
on	NN	O	O
IL-10	NN	O	B-protein
induced	NN	O	O
signal	NN	O	O
transduction	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
may	NN	O	O
help	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
intracellular	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
IL-10	NN	O	B-protein
stimulates	NN	O	O
human	NN	O	B-cell_type
B-cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Immunocytochemical	NN	O	O
analysis	NN	O	O
of	NN	O	O
MNDA	NN	O	B-protein
in	NN	O	O
tissue	NN	O	O
sections	NN	O	O
and	NN	O	O
sorted	NN	O	O
normal	NN	O	O
bone	NN	O	O
marrow	NN	O	O
cells	NN	O	O
documents	NN	O	O
expression	NN	O	O
only	NN	O	O
in	NN	O	O
maturing	NN	O	O
normal	NN	O	O
and	NN	O	O
neoplastic	NN	O	O
myelomonocytic	NN	O	O
cells	NN	O	O
and	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
normal	NN	O	O
and	NN	O	O
neoplastic	NN	O	O
B	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
differentiation	NN	O	O
antigen	NN	O	O
(	NN	O	O
MNDA	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
nuclear	NN	O	O
antigen	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
in	NN	O	O
mature	NN	O	O
myelomonocytic	NN	O	O
cell	NN	O	O
lines	NN	O	O
.	NN	O	O

An	NN	O	O
extensive	NN	O	O
immunocytochemical	NN	O	O
evaluation	NN	O	O
of	NN	O	O
fixed	NN	O	O
tissues	NN	O	O
confirmed	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
maturing	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
in	NN	O	O
acute	NN	O	O
nonlymphocytic	NN	O	O
leukemias	NN	O	O
and	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
.	NN	O	O

MNDA	NN	O	B-protein
was	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
normal	NN	O	O
tissue	NN	O	O
histiocytes	NN	O	O
but	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
foreign	NN	O	B-cell_type
body	NN	O	I-cell_type
giant	NN	O	I-cell_type
cells	NN	O	I-cell_type
associated	NN	O	O
with	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Flow	NN	O	O
cytometric	NN	O	O
cell	NN	O	O
sorting	NN	O	O
of	NN	O	O
normal	NN	O	O
bone	NN	O	O
marrow	NN	O	O
established	NN	O	O
that	NN	O	O
MNDA	NN	O	B-protein
is	NN	O	O
initially	NN	O	O
expressed	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
blast	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
showed	NN	O	O
a	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
in	NN	O	O
a	NN	O	O
population	NN	O	O
of	NN	O	O
normal	NN	O	O
mande	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
subset	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
neoplasms	NN	O	I-cell_type
expressing	NN	O	O
MNDA	NN	O	B-protein
included	NN	O	O
hairy	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
,	NN	O	O
parafollicular	NN	O	O
(	NN	O	O
monocytoid	NN	O	O
)	NN	O	O
B	NN	O	O
cell	NN	O	O
lymphoma	NN	O	O
,	NN	O	O
mantle	NN	O	O
cell	NN	O	O
lymphoma	NN	O	O
,	NN	O	O
and	NN	O	O
small	NN	O	O
lymphocytic	NN	O	O
lymphoma	NN	O	O
.	NN	O	O

Cell	NN	O	O
sorting	NN	O	O
of	NN	O	O
normal	NN	O	O
bone	NN	O	O
marrow	NN	O	O
showed	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
CD20	NN	O	B-protein
+/CD10-/CD5-	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

MNDA	NN	O	B-protein
was	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
other	NN	O	O
normal	NN	O	O
bone	NN	O	O
marrow	NN	O	O
or	NN	O	O
all	NN	O	O
other	NN	O	O
nonhematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
hematopoietic	NN	O	O
cell-specific	NN	O	O
pattern	NN	O	O
of	NN	O	O
MNDA	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
elucidated	NN	O	O
through	NN	O	O
a	NN	O	O
comprehensive	NN	O	O
analysis	NN	O	O
of	NN	O	O
normal	NN	O	O
and	NN	O	O
neoplastic	NN	O	O
tissues	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
results	NN	O	O
provide	NN	O	O
further	NN	O	O
evidence	NN	O	O
of	NN	O	O
the	NN	O	O
coexpression	NN	O	O
of	NN	O	O
B-	NN	O	B-protein
and	NN	O	I-protein
myeloid	NN	O	I-protein
cell	NN	O	I-protein
markers	NN	O	I-protein
in	NN	O	O
neoplastic	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
identify	NN	O	O
a	NN	O	O
normal	NN	O	B-cell_type
B	NN	O	I-cell_type
cell	NN	O	I-cell_type
population	NN	O	I-cell_type
that	NN	O	O
might	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
cell	NN	O	O
of	NN	O	O
origin	NN	O	O
of	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
neoplasms	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
activators	NN	O	I-protein
target	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
inhibit	NN	O	O
leukocyte	NN	O	B-cell_type
-endothelial	NN	O	B-cell_type
cell	NN	O	I-cell_type
interaction	NN	O	O
.	NN	O	O

An	NN	O	O
early	NN	O	O
event	NN	O	O
in	NN	O	O
acute	NN	O	O
and	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
and	NN	O	O
associated	NN	O	O
diseases	NN	O	O
such	NN	O	O
as	NN	O	O
atherosclerosis	NN	O	O
and	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
is	NN	O	O
the	NN	O	O
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
specific	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
ECs	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
which	NN	O	O
subsequently	NN	O	O
bind	NN	O	O
leukocytes	NN	O	B-cell_type
.	NN	O	O

Peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
PPARs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
are	NN	O	O
activated	NN	O	O
by	NN	O	O
fatty	NN	O	O
acid	NN	O	O
metabolites	NN	O	O
,	NN	O	O
peroxisome	NN	O	O
proliferators	NN	O	O
,	NN	O	O
and	NN	O	O
thiazolidinediones	NN	O	O
and	NN	O	O
are	NN	O	O
now	NN	O	O
recognized	NN	O	O
as	NN	O	O
important	NN	O	O
mediators	NN	O	O
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
.	NN	O	O

Whether	NN	O	O
PPAR	NN	O	O
activators	NN	O	O
influence	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
of	NN	O	O
ECs	NN	O	B-cell_type
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
PPAR	NN	O	O
activators	NN	O	O
15-deoxy-Delta	NN	O	O
(	NN	O	O
12	NN	O	O
,	NN	O	O
14	NN	O	O
)	NN	O	O
-prostaglandin	NN	O	O
J	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
(	NN	O	O
15d-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
)	NN	O	O
,	NN	O	O
Wyeth	NN	O	O
14643	NN	O	O
,	NN	O	O
ciglitazone	NN	O	O
,	NN	O	O
and	NN	O	O
troglitazone	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
BRL	NN	O	O
49653	NN	O	O
,	NN	O	O
partially	NN	O	O
inhibit	NN	O	O
the	NN	O	O
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
ELISA	NN	O	O
,	NN	O	O
and	NN	O	O
monocyte	NN	O	O
binding	NN	O	O
to	NN	O	O
human	NN	O	B-cell_type
aortic	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HAECs	NN	O	B-cell_type
)	NN	O	O
activated	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
or	NN	O	O
lipopolysaccharide	NN	O	O
.	NN	O	O

The	NN	O	O
``	NN	O	O
natural	NN	O	O
''	NN	O	O
PPAR	NN	O	O
activator	NN	O	O
15d-PGJ	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
had	NN	O	O
the	NN	O	O
greatest	NN	O	O
potency	NN	O	O
and	NN	O	O
was	NN	O	O
the	NN	O	O
only	NN	O	O
tested	NN	O	O
molecule	NN	O	O
capable	NN	O	O
of	NN	O	O
partially	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-protein
and	NN	O	O
neutrophil-like	NN	O	O
HL60	NN	O	B-cell_line
cell	NN	O	I-cell_line
binding	NN	O	O
to	NN	O	O
PMA-activated	NN	O	O
HAECs	NN	O	B-cell_type
.	NN	O	O

Intracellular	NN	O	O
adhesion	NN	O	O
molecule-1	NN	O	O
induction	NN	O	O
by	NN	O	O
PMA	NN	O	O
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
molecules	NN	O	O
tested	NN	O	O
.	NN	O	O

Both	NN	O	O
PPAR-alpha	NN	O	B-RNA
and	NN	O	I-RNA
PPAR-gamma	NN	O	I-RNA
mRNAs	NN	O	I-RNA
were	NN	O	O
detected	NN	O	O
in	NN	O	O
HAECs	NN	O	B-cell_type
by	NN	O	O
using	NN	O	O
reverse	NN	O	O
transcription-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
and	NN	O	O
a	NN	O	O
ribonuclease	NN	O	B-protein
protection	NN	O	O
assay	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
yet	NN	O	O
to	NN	O	O
determine	NN	O	O
which	NN	O	O
,	NN	O	O
if	NN	O	O
any	NN	O	O
,	NN	O	O
of	NN	O	O
the	NN	O	O
PPARs	NN	O	B-protein
are	NN	O	O
mediating	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
certain	NN	O	O
PPAR	NN	O	O
activators	NN	O	O
may	NN	O	O
help	NN	O	O
limit	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
mediated	NN	O	O
by	NN	O	O
VCAM-1	NN	O	B-protein
and	NN	O	O
monocytes	NN	O	B-cell_type
without	NN	O	O
affecting	NN	O	O
acute	NN	O	O
inflammation	NN	O	O
mediated	NN	O	O
by	NN	O	O
E-selectin	NN	O	B-protein
and	NN	O	O
neutrophil	NN	O	B-cell_type
binding	NN	O	O
.	NN	O	O

-DOCSTART-	O

Dephosphorylation	NN	O	O
of	NN	O	O
ZAP-70	NN	O	B-protein
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
by	NN	O	O
activated	NN	O	B-protein
SHP1	NN	O	I-protein
.	NN	O	O

Studies	NN	O	O
with	NN	O	O
motheaten	NN	O	O
mice	NN	O	O
,	NN	O	O
which	NN	O	O
lack	NN	O	O
the	NN	O	O
SHP1	NN	O	B-protein
protein	NN	O	I-protein
tyrosine	NN	O	B-protein
phosphatase	NN	O	I-protein
,	NN	O	O
indicate	NN	O	O
that	NN	O	O
this	NN	O	O
enzyme	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
negative	NN	O	O
role	NN	O	O
in	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
.	NN	O	O

The	NN	O	O
physiological	NN	O	O
substrates	NN	O	O
for	NN	O	O
SHP1	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
however	NN	O	O
,	NN	O	O
have	NN	O	O
remained	NN	O	O
unclear	NN	O	O
or	NN	O	O
controversial	NN	O	O
.	NN	O	O

To	NN	O	O
define	NN	O	O
these	NN	O	O
targets	NN	O	O
for	NN	O	O
SHP1	NN	O	B-protein
we	NN	O	O
have	NN	O	O
compared	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
constitutively	NN	O	O
active	NN	O	O
and	NN	O	O
inactive	NN	O	O
mutants	NN	O	O
of	NN	O	O
SHP1	NN	O	B-protein
on	NN	O	O
TCR	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
wild-type	NN	O	B-protein
SHP1	NN	O	I-protein
had	NN	O	O
a	NN	O	O
very	NN	O	O
small	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
TCR-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
ZAP-70	NN	O	B-protein
and	NN	O	O
Syk	NN	O	B-protein
,	NN	O	O
even	NN	O	O
when	NN	O	O
SHP1	NN	O	B-protein
was	NN	O	O
overexpressed	NN	O	O
20	NN	O	O
-	NN	O	O
100-fold	NN	O	O
over	NN	O	O
endogenous	NN	O	B-protein
SHP1	NN	O	I-protein
.	NN	O	O

Inactive	NN	O	B-protein
SHP1-D421A	NN	O	I-protein
and	NN	O	O
wild-type	NN	O	B-protein
SHP2	NN	O	I-protein
were	NN	O	O
without	NN	O	O
effects	NN	O	O
.	NN	O	O

Constitutively	NN	O	B-protein
active	NN	O	I-protein
SHP1-DeltaSH2	NN	O	I-protein
had	NN	O	O
a	NN	O	O
more	NN	O	O
pronounced	NN	O	O
effect	NN	O	O
on	NN	O	O
ZAP-70	NN	O	B-protein
and	NN	O	O
Syk	NN	O	B-protein
,	NN	O	O
even	NN	O	O
when	NN	O	O
expressed	NN	O	O
at	NN	O	O
near	NN	O	O
physiological	NN	O	O
levels	NN	O	O
.	NN	O	O

SHP1-DeltaSH2	NN	O	B-protein
also	NN	O	O
inhibited	NN	O	O
events	NN	O	O
downstream	NN	O	O
of	NN	O	O
ZAP-70	NN	O	B-protein
and	NN	O	O
Syk	NN	O	B-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
Erk2	NN	O	B-protein
and	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
constitutively	NN	O	O
active	NN	O	O
SHP2-DeltaSH2	NN	O	B-protein
had	NN	O	O
no	NN	O	O
statistically	NN	O	O
significant	NN	O	O
effect	NN	O	O
(	NN	O	O
although	NN	O	O
it	NN	O	O
caused	NN	O	O
a	NN	O	O
slight	NN	O	O
augmentation	NN	O	O
in	NN	O	O
some	NN	O	O
individual	NN	O	O
experiments	NN	O	O
)	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
constructs	NN	O	O
influenced	NN	O	O
the	NN	O	O
anti-CD3-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
zeta-chain	NN	O	I-protein
or	NN	O	O
phospholipase	NN	O	B-protein
Cgamma1	NN	O	I-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
Src	NN	O	B-protein
family	NN	O	I-protein
kinase	NN	O	I-protein
function	NN	O	O
was	NN	O	O
intact	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
support	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
ZAP-70	NN	O	B-protein
and	NN	O	O
Syk	NN	O	B-protein
can	NN	O	O
be	NN	O	O
direct	NN	O	O
substrates	NN	O	O
for	NN	O	O
SHP1	NN	O	B-protein
in	NN	O	O
intact	NN	O	O
cells	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
two	NN	O	O
SH2	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
SHP1	NN	O	B-protein
did	NN	O	O
not	NN	O	O
facilitate	NN	O	O
its	NN	O	O
recognition	NN	O	O
of	NN	O	O
ZAP-70	NN	O	B-protein
and	NN	O	O
Syk	NN	O	B-protein
as	NN	O	O
substrates	NN	O	O
in	NN	O	O
intact	NN	O	O
cells	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
suggest	NN	O	O
that	NN	O	O
SHP1	NN	O	B-protein
is	NN	O	O
not	NN	O	O
actively	NN	O	O
recruited	NN	O	O
to	NN	O	O
inhibit	NN	O	O
TCR	NN	O	B-protein
signaling	NN	O	O
induced	NN	O	O
by	NN	O	O
ligation	NN	O	O
of	NN	O	O
this	NN	O	O
receptor	NN	O	O
alone	NN	O	O
.	NN	O	O

Instead	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
ligation	NN	O	O
of	NN	O	O
a	NN	O	O
distinct	NN	O	O
inhibitory	NN	O	B-protein
receptor	NN	O	I-protein
leads	NN	O	O
to	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
SHP1	NN	O	B-protein
via	NN	O	O
its	NN	O	O
SH2	NN	O	B-protein
domains	NN	O	I-protein
,	NN	O	O
activation	NN	O	O
of	NN	O	O
SHP1	NN	O	B-protein
and	NN	O	O
subsequently	NN	O	O
inhibition	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
signals	NN	O	O
if	NN	O	O
the	NN	O	O
inhibitory	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
juxtaposed	NN	O	O
to	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Immune	NN	O	O
responses	NN	O	O
to	NN	O	O
adenoviruses	NN	O	O
:	NN	O	O
viral	NN	O	O
evasion	NN	O	O
mechanisms	NN	O	O
and	NN	O	O
their	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
clinic	NN	O	O
.	NN	O	O

Adenoviruses	NN	O	O
encode	NN	O	O
proteins	NN	O	O
that	NN	O	O
block	NN	O	O
responses	NN	O	O
to	NN	O	O
interferons	NN	O	B-protein
,	NN	O	O
intrinsic	NN	O	O
cellular	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
killing	NN	O	O
by	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cytotoxic	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
killing	NN	O	O
by	NN	O	O
the	NN	O	O
death	NN	O	B-protein
ligands	NN	O	I-protein
TNF	NN	O	B-protein
,	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
and	NN	O	O
TRAIL	NN	O	B-protein
.	NN	O	O

The	NN	O	O
viral	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
believed	NN	O	O
to	NN	O	O
prolong	NN	O	O
acute	NN	O	O
and	NN	O	O
persistent	NN	O	O
adenovirus	NN	O	O
infections	NN	O	O
.	NN	O	O

The	NN	O	O
proteins	NN	O	O
may	NN	O	O
prove	NN	O	O
useful	NN	O	O
in	NN	O	O
protecting	NN	O	O
adenovirus	NN	O	B-DNA
gene	NN	O	I-DNA
therapy	NN	O	I-DNA
vectors	NN	O	I-DNA
and	NN	O	O
transplanted	NN	O	B-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

-DOCSTART-	O

C/EBPbeta	NN	O	B-protein
and	NN	O	O
GATA-1	NN	O	B-protein
synergistically	NN	O	O
regulate	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
eosinophil	NN	O	B-DNA
granule	NN	O	I-DNA
major	NN	O	I-DNA
basic	NN	O	I-DNA
protein	NN	O	I-DNA
promoter	NN	O	I-DNA
:	NN	O	O
implication	NN	O	O
for	NN	O	O
C/EBPbeta	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
eosinophil	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Eosinophil	NN	O	B-protein
granule	NN	O	I-protein
major	NN	O	I-protein
basic	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
MBP	NN	O	B-protein
)	NN	O	O
is	NN	O	O
expressed	NN	O	O
exclusively	NN	O	O
in	NN	O	O
eosinophils	NN	O	B-cell_type
and	NN	O	O
basophils	NN	O	B-cell_type
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
our	NN	O	O
previous	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
major	NN	O	O
positive	NN	O	O
regulatory	NN	O	O
role	NN	O	O
for	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
a	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
role	NN	O	O
for	NN	O	O
GATA-2	NN	O	B-protein
in	NN	O	O
MBP	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

Further	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
MBP	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
identified	NN	O	O
a	NN	O	O
C/EBP	NN	O	B-DNA
(	NN	O	I-DNA
CCAAT/enhancer-binding	NN	O	I-DNA
protein	NN	O	I-DNA
)	NN	O	I-DNA
consensus	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
6	NN	O	O
bp	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
GATA-binding	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
MBP	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
HT93A	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
is	NN	O	O
capable	NN	O	O
of	NN	O	O
differentiating	NN	O	O
towards	NN	O	O
both	NN	O	O
the	NN	O	O
eosinophil	NN	O	B-cell_type
and	NN	O	O
neutrophil	NN	O	O
lineages	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
,	NN	O	O
C/EBPalpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
decreased	NN	O	O
significantly	NN	O	O
concomitant	NN	O	O
with	NN	O	O
eosinophilic	NN	O	O
and	NN	O	O
neutrophilic	NN	O	O
differentiation	NN	O	O
,	NN	O	O
whereas	NN	O	O
C/EBPbeta	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
markedly	NN	O	O
increased	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSAs	NN	O	O
)	NN	O	O
showed	NN	O	O
that	NN	O	O
recombinant	NN	O	O
C/EBPbeta	NN	O	B-protein
protein	NN	O	O
could	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
potential	NN	O	O
C/EBP-binding	NN	O	B-DNA
site	NN	O	I-DNA
(	NN	O	O
bp	NN	O	B-DNA
-90	NN	O	I-DNA
to	NN	O	I-DNA
-82	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
MBP	NN	O	B-protein
promoter	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
both	NN	O	O
C/EBPbeta	NN	O	B-protein
and	NN	O	O
GATA-1	NN	O	B-protein
can	NN	O	O
bind	NN	O	O
simultaneously	NN	O	O
to	NN	O	O
the	NN	O	O
C/EBP-	NN	O	O
and	NN	O	O
GATA-binding	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
MBP	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
functionality	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
C/EBP-	NN	O	B-DNA
and	NN	O	I-DNA
GATA-	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
whether	NN	O	O
C/EBPbeta	NN	O	B-protein
and	NN	O	O
GATA-1	NN	O	B-protein
can	NN	O	O
stimulate	NN	O	O
the	NN	O	O
MBP	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
C/EBPbeta	NN	O	B-cell_line
and	NN	O	I-cell_line
GATA-1	NN	O	I-cell_line
negative	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Cotransfection	NN	O	O
with	NN	O	O
C/EBPbeta	NN	O	B-DNA
and	NN	O	I-DNA
GATA-1	NN	O	I-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
produced	NN	O	O
a	NN	O	O
5-fold	NN	O	O
increase	NN	O	O
compared	NN	O	O
with	NN	O	O
cotransfection	NN	O	O
with	NN	O	O
the	NN	O	O
C/EBPbeta	NN	O	B-protein
or	NN	O	O
GATA-1	NN	O	B-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
individually	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
GST	NN	O	B-protein
pull-down	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
physical	NN	O	O
interaction	NN	O	O
between	NN	O	O
human	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
C/EBPbeta	NN	O	B-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
FOG	NN	O	B-protein
(	NN	O	O
riend	NN	O	B-protein
ATA	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
cofactor	NN	O	O
for	NN	O	O
GATA-binding	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
decreased	NN	O	O
transactivation	NN	O	O
activity	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
for	NN	O	O
the	NN	O	O
MBP	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
provide	NN	O	O
the	NN	O	O
first	NN	O	O
evidence	NN	O	O
that	NN	O	O
both	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
C/EBPbeta	NN	O	B-protein
synergistically	NN	O	O
transactivate	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
an	NN	O	O
eosinophil-specific	NN	O	B-DNA
granule	NN	O	I-DNA
protein	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
FOG	NN	O	B-protein
may	NN	O	O
act	NN	O	O
as	NN	O	O
a	NN	O	O
negative	NN	O	O
cofactor	NN	O	O
for	NN	O	O
the	NN	O	O
eosinophil	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
unlike	NN	O	O
its	NN	O	O
positively	NN	O	O
regulatory	NN	O	O
function	NN	O	O
for	NN	O	O
the	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
megakaryocyte	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Tissue-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
ecto-5'-nucleotidase	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Role	NN	O	O
of	NN	O	O
the	NN	O	O
camp	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
mediating	NN	O	O
repression	NN	O	O
by	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
isolated	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
ecto-5'-nucleotidase	NN	O	B-DNA
(	NN	O	I-DNA
low	NN	O	I-DNA
K	NN	O	I-DNA
(	NN	O	I-DNA
m	NN	O	I-DNA
)	NN	O	I-DNA
5'-NT	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
established	NN	O	O
that	NN	O	O
a	NN	O	O
969-base	NN	O	B-DNA
pair	NN	O	I-DNA
(	NN	O	I-DNA
bp	NN	O	I-DNA
)	NN	O	I-DNA
fragment	NN	O	I-DNA
confers	NN	O	O
cell-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
that	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
endogenous	NN	O	B-RNA
ecto-5'-NT	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
.	NN	O	O

A	NN	O	O
768-bp	NN	O	B-DNA
upstream	NN	O	I-DNA
negative	NN	O	I-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
identified	NN	O	O
that	NN	O	O
conferred	NN	O	O
lymphocyte-specific	NN	O	O
negative	NN	O	O
regulation	NN	O	O
in	NN	O	O
a	NN	O	O
heterologous	NN	O	O
system	NN	O	O
with	NN	O	O
a	NN	O	O
244-bp	NN	O	B-DNA
deoxycytidine	NN	O	I-DNA
kinase	NN	O	I-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

DNase	NN	O	B-protein
I	NN	O	I-protein
footprinting	NN	O	O
identified	NN	O	O
several	NN	O	O
protected	NN	O	O
areas	NN	O	O
including	NN	O	O
Sp1	NN	O	B-DNA
,	NN	O	I-DNA
Sp1/AP-2	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
cAMP	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
CRE	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
201-bp	NN	O	O
core	NN	O	O
promoter	NN	O	O
region	NN	O	O
and	NN	O	O
Sp1	NN	O	B-DNA
,	NN	O	I-DNA
NRE-2a	NN	O	I-DNA
,	NN	O	I-DNA
TCF-1/LEF-1	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
Sp1/NF-AT	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Whereas	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
was	NN	O	O
essential	NN	O	O
in	NN	O	O
mediating	NN	O	O
the	NN	O	O
negative	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
Sp1/AP-2	NN	O	B-DNA
site	NN	O	I-DNA
decreased	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
lines	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
analysis	NN	O	O
of	NN	O	O
proteins	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
identified	NN	O	O
both	NN	O	O
ATF-1	NN	O	B-protein
and	NN	O	O
ATF-2	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
increased	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
core	NN	O	O
and	NN	O	O
the	NN	O	O
969-bp	NN	O	B-DNA
promoter	NN	O	I-DNA
fragments	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
this	NN	O	O
increase	NN	O	O
was	NN	O	O
abrogated	NN	O	O
by	NN	O	O
mutations	NN	O	O
at	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
tissue-specific	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
5	NN	O	O
'	NN	O	O
of	NN	O	O
the	NN	O	O
ecto-5'-NT	NN	O	B-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
that	NN	O	O
requires	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
functional	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
basal	NN	O	B-DNA
promoter	NN	O	I-DNA
for	NN	O	O
its	NN	O	O
suppressive	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Extracellular	NN	O	O
signal-regulated	NN	O	O
protein	NN	O	O
kinase	NN	O	O
(	NN	O	O
ERK	NN	O	O
)	NN	O	O
-dependent	NN	O	O
and	NN	O	O
ERK-independent	NN	O	O
pathways	NN	O	O
target	NN	O	O
STAT3	NN	O	B-protein
on	NN	O	O
serine-727	NN	O	O
in	NN	O	O
human	NN	O	O
neutrophils	NN	O	O
stimulated	NN	O	O
by	NN	O	O
chemotactic	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

STAT3	NN	O	B-protein
(	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
3	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
latent	NN	O	O
transcription	NN	O	O
factor	NN	O	O
that	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
(	NN	O	O
Tyr-705	NN	O	O
)	NN	O	O
in	NN	O	O
cells	NN	O	O
stimulated	NN	O	O
with	NN	O	O
cytokines	NN	O	B-protein
or	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
cytoplasmic	NN	O	B-protein
serine	NN	O	I-protein
kinases	NN	O	I-protein
also	NN	O	O
phosphorylate	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
are	NN	O	O
necessary	NN	O	O
for	NN	O	O
maximal	NN	O	O
gene	NN	O	O
activation	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
site-specific	NN	O	O
antibody	NN	O	O
,	NN	O	O
that	NN	O	O
STAT3	NN	O	B-protein
is	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
Ser-727	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
chemotactic	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
N-formyl-methionyl-leucyl-phenylalanine	NN	O	O
and	NN	O	O
complement	NN	O	O
C5a	NN	O	O
)	NN	O	O
,	NN	O	O
cytokines	NN	O	B-protein
[	NN	O	O
granulocyte/macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
G-CSF	NN	O	B-protein
)	NN	O	O
]	NN	O	O
,	NN	O	O
or	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
activator	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
.	NN	O	O

(	NN	O	O
2-Amino-3'-methoxyphenyl	NN	O	O
)	NN	O	O
oxanaphthalen-4-one	NN	O	O
(	NN	O	O
PD	NN	O	O
98059	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
extracellular	NN	O	O
signal-regulated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
ERK	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
,	NN	O	O
blocked	NN	O	O
the	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
induced	NN	O	O
by	NN	O	O
chemotactic	NN	O	B-protein
factors	NN	O	I-protein
or	NN	O	O
PMA	NN	O	O
.	NN	O	O

The	NN	O	O
drug	NN	O	O
was	NN	O	O
less	NN	O	O
effective	NN	O	O
on	NN	O	O
cytokines	NN	O	B-protein
:	NN	O	O
it	NN	O	O
virtually	NN	O	O
abolished	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
GM-CSF	NN	O	B-protein
that	NN	O	O
occurred	NN	O	O
5	NN	O	O
min	NN	O	O
after	NN	O	O
stimulation	NN	O	O
but	NN	O	O
only	NN	O	O
partly	NN	O	O
decreased	NN	O	O
those	NN	O	O
at	NN	O	O
15-30	NN	O	O
min	NN	O	O
and	NN	O	O
did	NN	O	O
not	NN	O	O
appreciably	NN	O	O
alter	NN	O	O
responses	NN	O	O
to	NN	O	O
G-CSF	NN	O	B-protein
regardless	NN	O	O
of	NN	O	O
incubation	NN	O	O
time	NN	O	O
.	NN	O	O

1-	NN	O	O
(	NN	O	O
5-Isoquinolinylsulphonyl	NN	O	O
)	NN	O	O
-2-methylpiperazine	NN	O	O
dihydrochloride	NN	O	O
(	NN	O	O
H7	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
a	NN	O	O
putative	NN	O	O
STAT3	NN	O	B-protein
serine	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
4-	NN	O	O
(	NN	O	O
4-fluorophenyl	NN	O	O
)	NN	O	O
-2-	NN	O	O
(	NN	O	O
4-methylsulphinylphenyl	NN	O	O
)	NN	O	O
-5-	NN	O	O
(	NN	O	O
4-pyridyl	NN	O	O
)	NN	O	O
1H-imidazole	NN	O	O
(	NN	O	O
SB	NN	O	O
203580	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MAP	NN	O	I-protein
)	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
did	NN	O	O
not	NN	O	O
dampen	NN	O	O
any	NN	O	O
of	NN	O	O
these	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
responses	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
neutrophils	NN	O	B-cell_type
use	NN	O	O
both	NN	O	O
ERK-dependent	NN	O	O
and	NN	O	O
ERK-independent	NN	O	O
pathways	NN	O	O
to	NN	O	O
phosphorylate	NN	O	O
Ser-727	NN	O	O
on	NN	O	O
STAT3	NN	O	B-protein
.	NN	O	O

The	NN	O	O
former	NN	O	O
pathway	NN	O	O
is	NN	O	O
recruited	NN	O	O
by	NN	O	O
all	NN	O	O
ERK	NN	O	B-protein
-activating	NN	O	O
stimuli	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
latter	NN	O	O
pathway	NN	O	O
uses	NN	O	O
an	NN	O	O
undefined	NN	O	O
serine	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
is	NN	O	O
recruited	NN	O	O
selectively	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

GATA	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
associate	NN	O	O
with	NN	O	O
a	NN	O	O
novel	NN	O	O
class	NN	O	O
of	NN	O	O
nuclear	NN	O	O
bodies	NN	O	O
in	NN	O	O
erythroblasts	NN	O	B-cell_type
and	NN	O	O
megakaryocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
nuclear	NN	O	O
distribution	NN	O	O
of	NN	O	O
GATA	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
murine	NN	O	B-cell_type
haemopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
examined	NN	O	O
by	NN	O	O
indirect	NN	O	O
immunofluorescence	NN	O	O
.	NN	O	O

Specific	NN	O	O
bright	NN	O	O
foci	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
fluorescence	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
erythroleukaemia	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
primary	NN	O	B-cell_type
murine	NN	O	I-cell_type
erythroblasts	NN	O	I-cell_type
and	NN	O	O
megakaryocytes	NN	O	B-cell_type
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
diffuse	NN	O	O
nucleoplasmic	NN	O	O
localization	NN	O	O
.	NN	O	O

These	NN	O	O
foci	NN	O	O
,	NN	O	O
which	NN	O	O
were	NN	O	O
preferentially	NN	O	O
found	NN	O	O
adjacent	NN	O	O
to	NN	O	O
nucleoli	NN	O	O
or	NN	O	O
at	NN	O	O
the	NN	O	O
nuclear	NN	O	O
periphery	NN	O	O
,	NN	O	O
did	NN	O	O
not	NN	O	O
represent	NN	O	O
sites	NN	O	O
of	NN	O	O
active	NN	O	O
transcription	NN	O	O
or	NN	O	O
binding	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
to	NN	O	O
consensus	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
loci	NN	O	I-DNA
.	NN	O	O

Immunoelectron	NN	O	O
microscopy	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
intensely	NN	O	O
labelled	NN	O	O
structures	NN	O	O
likely	NN	O	O
to	NN	O	O
represent	NN	O	O
the	NN	O	O
GATA-1	NN	O	B-protein
foci	NN	O	O
seen	NN	O	O
by	NN	O	O
immunofluorescence	NN	O	O
.	NN	O	O

The	NN	O	O
GATA-1	NN	O	B-protein
nuclear	NN	O	O
bodies	NN	O	O
differed	NN	O	O
from	NN	O	O
previously	NN	O	O
described	NN	O	O
nuclear	NN	O	O
structures	NN	O	O
and	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
co-localization	NN	O	O
with	NN	O	O
nuclear	NN	O	B-protein
antigens	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
RNA	NN	O	O
processing	NN	O	O
or	NN	O	O
other	NN	O	O
ubiquitous	NN	O	B-protein
(	NN	O	I-protein
Spl	NN	O	I-protein
,	NN	O	I-protein
c-Jun	NN	O	I-protein
and	NN	O	I-protein
TBP	NN	O	I-protein
)	NN	O	I-protein
or	NN	O	I-protein
haemopoietic	NN	O	I-protein
(	NN	O	I-protein
NF-E2	NN	O	I-protein
)	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
GATA-2	NN	O	B-protein
and	NN	O	O
GATA-3	NN	O	B-protein
proteins	NN	O	I-protein
also	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
nuclear	NN	O	O
bodies	NN	O	O
in	NN	O	O
cell	NN	O	O
lines	NN	O	O
co-expressing	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	I-protein
-2	NN	O	I-protein
or	NN	O	I-protein
GATA-1	NN	O	I-protein
and	NN	O	I-protein
-3	NN	O	I-protein
gene	NN	O	I-protein
products	NN	O	I-protein
.	NN	O	O

This	NN	O	O
pattern	NN	O	O
of	NN	O	O
distribution	NN	O	O
is	NN	O	O
,	NN	O	O
thus	NN	O	O
far	NN	O	O
,	NN	O	O
unique	NN	O	O
to	NN	O	O
the	NN	O	O
GATA	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
and	NN	O	O
suggests	NN	O	O
a	NN	O	O
protein-protein	NN	O	O
interaction	NN	O	O
with	NN	O	O
other	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
bodies	NN	O	O
via	NN	O	O
the	NN	O	O
GATA	NN	O	B-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Energy	NN	O	O
substrates	NN	O	O
,	NN	O	O
hormone	NN	O	O
responses	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
during	NN	O	O
intense	NN	O	O
physical	NN	O	O
exercise	NN	O	O
in	NN	O	O
humans	NN	O	O
following	NN	O	O
phosphocreatine	NN	O	O
administration	NN	O	O
.	NN	O	O

Eight	NN	O	O
healthy	NN	O	O
untrained	NN	O	O
male	NN	O	O
volunteers	NN	O	O
pedalled	NN	O	O
a	NN	O	O
cycle	NN	O	O
ergometer	NN	O	O
according	NN	O	O
to	NN	O	O
two	NN	O	O
exercise	NN	O	O
protocols	NN	O	O
:	NN	O	O
the	NN	O	O
first	NN	O	O
involved	NN	O	O
step-wise	NN	O	O
increasing	NN	O	O
physical	NN	O	O
exercise	NN	O	O
to	NN	O	O
maximal	NN	O	O
(	NN	O	O
MPE	NN	O	O
)	NN	O	O
;	NN	O	O
the	NN	O	O
second	NN	O	O
involved	NN	O	O
prolonged	NN	O	O
(	NN	O	O
35	NN	O	O
min	NN	O	O
)	NN	O	O
submaximal	NN	O	O
physical	NN	O	O
exercise	NN	O	O
(	NN	O	O
PPE	NN	O	O
)	NN	O	O
at	NN	O	O
70	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
individual	NN	O	O
's	NN	O	O
maximal	NN	O	O
oxygen	NN	O	O
uptake	NN	O	O
.	NN	O	O

Each	NN	O	O
volunteer	NN	O	O
performed	NN	O	O
these	NN	O	O
exercise	NN	O	O
twice	NN	O	O
,	NN	O	O
following	NN	O	O
either	NN	O	O
an	NN	O	O
intravenous	NN	O	O
injection	NN	O	O
of	NN	O	O
phosphocreatine	NN	O	O
(	NN	O	O
PCr	NN	O	O
)	NN	O	O
or	NN	O	O
a	NN	O	O
placebo	NN	O	O
of	NN	O	O
an	NN	O	O
isotonic	NN	O	O
NaCl	NN	O	O
solution	NN	O	O
.	NN	O	O

Anaerobic	NN	O	O
threshold	NN	O	O
(	NN	O	O
AT	NN	O	O
)	NN	O	O
was	NN	O	O
determined	NN	O	O
from	NN	O	O
the	NN	O	O
point	NN	O	O
of	NN	O	O
departure	NN	O	O
of	NN	O	O
the	NN	O	O
ventilatory	NN	O	O
response	NN	O	O
from	NN	O	O
linearity	NN	O	O
and	NN	O	O
from	NN	O	O
the	NN	O	O
sudden	NN	O	O
increase	NN	O	O
in	NN	O	O
venous	NN	O	O
blood	NN	O	O
lactate	NN	O	O
concentrations	NN	O	O
during	NN	O	O
MPE	NN	O	O
.	NN	O	O

After	NN	O	O
exercise	NN	O	O
following	NN	O	O
placebo	NN	O	O
administration	NN	O	O
we	NN	O	O
observed	NN	O	O
increases	NN	O	O
in	NN	O	O
concentrations	NN	O	O
of	NN	O	O
blood	NN	O	O
substrates	NN	O	O
,	NN	O	O
plasma	NN	O	B-protein
adrenocorticotropin	NN	O	I-protein
(	NN	O	O
ACTH	NN	O	B-protein
)	NN	O	O
,	NN	O	O
growth	NN	O	O
hormone	NN	O	O
and	NN	O	O
cortisol	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
without	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
.	NN	O	O

Intravenous	NN	O	O
administration	NN	O	O
of	NN	O	O
PCr	NN	O	O
(	NN	O	O
starting	NN	O	O
1	NN	O	O
day	NN	O	O
before	NN	O	O
exercise	NN	O	O
)	NN	O	O
led	NN	O	O
to	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
total	NN	O	O
workload	NN	O	O
(	NN	O	O
on	NN	O	O
average	NN	O	O
by	NN	O	O
5.8	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
AT	NN	O	O
(	NN	O	O
on	NN	O	O
average	NN	O	O
by	NN	O	O
6.8	NN	O	O
%	NN	O	O
)	NN	O	O
during	NN	O	O
MPE	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
better	NN	O	O
tolerance	NN	O	O
of	NN	O	O
exercise	NN	O	O
during	NN	O	O
PPE	NN	O	O
.	NN	O	O

Following	NN	O	O
PCr	NN	O	O
administration	NN	O	O
we	NN	O	O
observed	NN	O	O
lower	NN	O	O
blood	NN	O	O
lactate	NN	O	O
concentrations	NN	O	O
and	NN	O	O
different	NN	O	O
patterns	NN	O	O
of	NN	O	O
some	NN	O	O
enzyme	NN	O	O
activities	NN	O	O
,	NN	O	O
less	NN	O	O
pronounced	NN	O	O
changes	NN	O	O
in	NN	O	O
plasma	NN	O	B-protein
ACTH	NN	O	I-protein
and	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
and	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
no	NN	O	O
changes	NN	O	O
in	NN	O	O
plasma	NN	O	O
growth	NN	O	O
hormone	NN	O	O
concentrations	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
placebo	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
intense	NN	O	O
physical	NN	O	O
exercise	NN	O	O
led	NN	O	O
not	NN	O	O
only	NN	O	O
to	NN	O	O
increases	NN	O	O
in	NN	O	O
blood	NN	O	O
hormone	NN	O	O
concentrations	NN	O	O
but	NN	O	O
also	NN	O	O
to	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
density	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Intravenous	NN	O	O
PCr	NN	O	O
injection	NN	O	O
led	NN	O	O
to	NN	O	O
smaller	NN	O	O
changes	NN	O	O
in	NN	O	O
ACTH	NN	O	B-protein
and	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
to	NN	O	O
a	NN	O	O
lower	NN	O	O
activation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Putative	NN	O	B-protein
steroid	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
mineralocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
expressed	NN	O	O
in	NN	O	O
E.	NN	O	O
coli	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
proteins	NN	O	I-protein
shows	NN	O	O
typical	NN	O	O
native	NN	O	O
receptor	NN	O	O
characteristics	NN	O	O
.	NN	O	O

Domain	NN	O	B-protein
E	NN	O	I-protein
,	NN	O	O
considered	NN	O	O
as	NN	O	O
the	NN	O	O
putative	NN	O	B-protein
hormone	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
(	NN	O	O
HBD	NN	O	B-protein
)	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
mineralocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
hMR	NN	O	B-protein
)	NN	O	O
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
as	NN	O	O
a	NN	O	O
fusion	NN	O	O
protein	NN	O	O
with	NN	O	O
either	NN	O	O
maltose	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
MBP	NN	O	B-protein
)	NN	O	O
or	NN	O	O
glutathione	NN	O	B-protein
S-transferase	NN	O	I-protein
(	NN	O	O
GST	NN	O	B-protein
)	NN	O	O
.	NN	O	O

These	NN	O	O
bacterially-produced	NN	O	B-DNA
MR	NN	O	I-DNA
constructs	NN	O	I-DNA
had	NN	O	O
no	NN	O	O
steroid	NN	O	O
binding	NN	O	O
activity	NN	O	O
per	NN	O	O
se	NN	O	O
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
protein	NN	O	I-protein
association	NN	O	O
(	NN	O	O
hsp	NN	O	B-protein
)	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
high	NN	O	O
affinity	NN	O	O
ligand-binding	NN	O	O
of	NN	O	O
the	NN	O	O
MR	NN	O	B-protein
.	NN	O	O

After	NN	O	O
incubation	NN	O	O
of	NN	O	O
purified	NN	O	O
MBP-	NN	O	B-protein
or	NN	O	I-protein
GST-HBD	NN	O	I-protein
with	NN	O	O
rabbit	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
,	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
rich	NN	O	O
in	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
we	NN	O	O
obtained	NN	O	O
saturable	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
aldosterone	NN	O	O
.	NN	O	O

The	NN	O	O
Kd	NN	O	O
value	NN	O	O
for	NN	O	O
aldosterone	NN	O	O
was	NN	O	O
0.3	NN	O	O
nM	NN	O	O
and	NN	O	O
the	NN	O	O
Bmax	NN	O	O
=	NN	O	O
32	NN	O	O
pmol/mg	NN	O	O
.	NN	O	O

Hormone	NN	O	O
binding	NN	O	O
specificity	NN	O	O
was	NN	O	O
assessed	NN	O	O
by	NN	O	O
competition	NN	O	O
studies	NN	O	O
with	NN	O	O
various	NN	O	O
steroid	NN	O	O
ligands	NN	O	O
.	NN	O	O

Sucrose	NN	O	O
gradient	NN	O	O
assays	NN	O	O
performed	NN	O	O
with	NN	O	O
[	NN	O	B-protein
3H	NN	O	I-protein
]	NN	O	I-protein
aldosterone-MBP-HBD	NN	O	I-protein
revealed	NN	O	O
complex	NN	O	O
sedimenting	NN	O	O
at	NN	O	O
8.3S	NN	O	O
and	NN	O	O
4.9S	NN	O	O
with	NN	O	O
[	NN	O	B-protein
3H	NN	O	I-protein
]	NN	O	I-protein
progesterone-MBP-HBD	NN	O	I-protein
.	NN	O	O

Western-blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
sedimentation	NN	O	O
peak	NN	O	O
showed	NN	O	O
the	NN	O	O
concomitant	NN	O	O
presence	NN	O	O
of	NN	O	O
MBP-HBD	NN	O	B-protein
by	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
anti-MBP	NN	O	I-protein
antibody	NN	O	I-protein
,	NN	O	O
and	NN	O	O
hsp90	NN	O	B-protein
by	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
anti-hsp	NN	O	I-protein
antibody	NN	O	I-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
following	NN	O	O
incubation	NN	O	O
with	NN	O	O
the	NN	O	O
anti-rabbit	NN	O	B-protein
hsp90	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
the	NN	O	O
sedimenting	NN	O	O
gradient	NN	O	O
showed	NN	O	O
a	NN	O	O
10.4S	NN	O	B-protein
sedimenting	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

These	NN	O	O
analyses	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
aldosterone-	NN	O	O
MBP-HBD	NN	O	B-protein
complex	NN	O	I-protein
is	NN	O	O
at	NN	O	O
least	NN	O	O
associated	NN	O	O
with	NN	O	O
hsp90	NN	O	B-protein
in	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
HBD	NN	O	B-protein
of	NN	O	O
hMR	NN	O	B-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
bind	NN	O	O
hsp90	NN	O	B-protein
.	NN	O	O

Deletions	NN	O	O
of	NN	O	O
a	NN	O	O
relatively	NN	O	O
short	NN	O	O
amino-	NN	O	B-protein
(	NN	O	I-protein
729-766	NN	O	I-protein
)	NN	O	I-protein
or	NN	O	I-protein
carboxy-	NN	O	I-protein
terminal	NN	O	I-protein
(	NN	O	I-protein
940-984	NN	O	I-protein
)	NN	O	I-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HBD	NN	O	B-protein
fragment	NN	O	I-protein
eliminated	NN	O	O
all	NN	O	O
steroid-binding	NN	O	O
properties	NN	O	O
.	NN	O	O

Overall	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
integrity	NN	O	O
of	NN	O	O
domain	NN	O	B-protein
E	NN	O	I-protein
is	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
to	NN	O	O
bind	NN	O	O
steroid	NN	O	O
ligands	NN	O	O
,	NN	O	O
agonists	NN	O	O
or	NN	O	O
antagonists	NN	O	O
,	NN	O	O
with	NN	O	O
characteristics	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
entire	NN	O	O
native	NN	O	O
MR	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Renal	NN	O	O
cell	NN	O	O
carcinoma-derived	NN	O	O
gangliosides	NN	O	O
suppress	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NFkappaB	NN	O	B-protein
)	NN	O	O
is	NN	O	O
impaired	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
renal	NN	O	O
cell	NN	O	O
carcinomas	NN	O	O
(	NN	O	O
RCCs	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
circulating	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
RCCs	NN	O	O
,	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
downstream	NN	O	O
from	NN	O	O
the	NN	O	O
stimulus-induced	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
factor	NN	O	I-protein
IkappaBalpha	NN	O	B-protein
.	NN	O	O

Tumor-derived	NN	O	B-protein
soluble	NN	O	I-protein
products	NN	O	I-protein
from	NN	O	O
cultured	NN	O	O
RCC	NN	O	O
explants	NN	O	O
inhibit	NN	O	O
NFkappaB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
healthy	NN	O	O
volunteers	NN	O	O
,	NN	O	O
despite	NN	O	O
a	NN	O	O
normal	NN	O	O
level	NN	O	O
of	NN	O	O
stimulus-induced	NN	O	O
IkappaBalpha	NN	O	B-protein
degradation	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
agent	NN	O	O
has	NN	O	O
several	NN	O	O
features	NN	O	O
characteristic	NN	O	O
of	NN	O	O
a	NN	O	O
ganglioside	NN	O	O
,	NN	O	O
including	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
neuraminidase	NN	O	B-protein
but	NN	O	O
not	NN	O	O
protease	NN	O	B-protein
treatment	NN	O	O
;	NN	O	O
hydrophobicity	NN	O	O
;	NN	O	O
and	NN	O	O
molecular	NN	O	O
weight	NN	O	O
less	NN	O	O
than	NN	O	O
3	NN	O	O
kDa	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
we	NN	O	O
detected	NN	O	O
gangliosides	NN	O	O
in	NN	O	O
supernatants	NN	O	O
from	NN	O	O
RCC	NN	O	B-cell_type
explants	NN	O	I-cell_type
and	NN	O	O
not	NN	O	O
from	NN	O	O
adjacent	NN	O	O
normal	NN	O	O
kidney	NN	O	O
tissue	NN	O	O
.	NN	O	O

Gangliosides	NN	O	O
prepared	NN	O	O
from	NN	O	O
RCC	NN	O	O
supernatants	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
purified	NN	O	O
bovine	NN	O	O
gangliosides	NN	O	O
G	NN	O	O
(	NN	O	O
m1	NN	O	O
)	NN	O	O
and	NN	O	O
G	NN	O	O
(	NN	O	O
d1a	NN	O	O
)	NN	O	O
,	NN	O	O
suppressed	NN	O	O
NFkappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
reduced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
IL-2	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
tumor-derived	NN	O	O
gangliosides	NN	O	O
may	NN	O	O
blunt	NN	O	O
antitumor	NN	O	O
immune	NN	O	O
responses	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
RCCs	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
-dependent	NN	O	O
induction	NN	O	O
of	NN	O	O
interleukin-8	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
antioxidant	NN	O	O
sensitive	NN	O	O
activating	NN	O	O
pathway	NN	O	O
distinct	NN	O	O
from	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNFalpha	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
pluripotent	NN	O	O
activator	NN	O	O
of	NN	O	O
inflammation	NN	O	O
by	NN	O	O
inducing	NN	O	O
a	NN	O	O
proinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
cascade	NN	O	O
.	NN	O	O

This	NN	O	O
phenomenon	NN	O	O
is	NN	O	O
mediated	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
through	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CXC	NN	O	B-protein
chemokine	NN	O	I-protein
,	NN	O	O
interleukin-8	NN	O	B-protein
(	NN	O	O
IL-8	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
TNFalpha	NN	O	B-protein
-inducible	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
in	NN	O	O
IL-8	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
``	NN	O	O
monocyte-like	NN	O	O
''	NN	O	O
U937	NN	O	B-cell_line
histiocytic	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

TNFalpha	NN	O	B-protein
is	NN	O	O
a	NN	O	O
rapid	NN	O	O
activator	NN	O	O
of	NN	O	O
IL-8	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
U937	NN	O	B-cell_line
,	NN	O	O
producing	NN	O	O
a	NN	O	O
50-fold	NN	O	O
induction	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
within	NN	O	O
1	NN	O	O
hour	NN	O	O
of	NN	O	O
treatment	NN	O	O
.	NN	O	O

In	NN	O	O
gene	NN	O	O
transfection	NN	O	O
assays	NN	O	O
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
TNFalpha	NN	O	B-protein
requires	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	B-DNA
nuclear	NN	O	I-DNA
factor-kappaB	NN	O	I-DNA
(	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
Rel	NN	O	I-DNA
A	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-8	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

TNFalpha	NN	O	B-protein
treatment	NN	O	O
induces	NN	O	O
a	NN	O	O
rapid	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
65	NN	O	B-protein
kD	NN	O	I-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
NF-kappaB	NN	O	B-protein
subunit	NN	O	I-protein
,	NN	O	O
Rel	NN	O	B-protein
A	NN	O	I-protein
,	NN	O	O
whose	NN	O	O
binding	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
occurs	NN	O	O
before	NN	O	O
changes	NN	O	O
in	NN	O	O
intracellular	NN	O	O
ROS	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
(	NN	O	O
or	NN	O	O
up	NN	O	O
to	NN	O	O
15	NN	O	O
minutes	NN	O	O
posttreatment	NN	O	O
)	NN	O	O
relative	NN	O	O
to	NN	O	O
TNFalpha	NN	O	B-protein
with	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
dimethyl	NN	O	O
sulfoxide	NN	O	O
(	NN	O	O
DMSO	NN	O	O
)	NN	O	O
(	NN	O	O
2	NN	O	O
%	NN	O	O
[	NN	O	O
vol/vol	NN	O	O
]	NN	O	O
)	NN	O	O
blocks	NN	O	O
80	NN	O	O
%	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
DMSO	NN	O	O
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
inducible	NN	O	O
Rel	NN	O	B-protein
A	NN	O	I-protein
binding	NN	O	O
.	NN	O	O

Similar	NN	O	O
selective	NN	O	O
effects	NN	O	O
on	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	O
are	NN	O	O
seen	NN	O	O
with	NN	O	O
the	NN	O	O
unrelated	NN	O	O
antioxidants	NN	O	O
,	NN	O	O
N-acetylcysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
and	NN	O	O
vitamin	NN	O	O
C	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
TNFalpha	NN	O	B-protein
induces	NN	O	O
a	NN	O	O
delayed	NN	O	O
ROS-dependent	NN	O	O
signalling	NN	O	O
pathway	NN	O	O
that	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
transcriptional	NN	O	O
activation	NN	O	O
and	NN	O	O
is	NN	O	O
separable	NN	O	O
from	NN	O	O
that	NN	O	O
required	NN	O	O
for	NN	O	O
its	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

Further	NN	O	O
definition	NN	O	O
of	NN	O	O
this	NN	O	O
pathway	NN	O	O
will	NN	O	O
yield	NN	O	O
new	NN	O	O
insights	NN	O	O
into	NN	O	O
inflammation	NN	O	O
initiated	NN	O	O
by	NN	O	O
TNFalpha	NN	O	B-protein
signalling	NN	O	O
.	NN	O	O

-DOCSTART-	O

Gender	NN	O	O
and	NN	O	O
vascular	NN	O	O
reactivity	NN	O	O
.	NN	O	O

Estrogen	NN	O	B-protein
receptors	NN	O	I-protein
are	NN	O	O
found	NN	O	O
on	NN	O	O
vascular	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
and	NN	O	I-cell_type
smooth	NN	O	I-cell_type
muscle	NN	O	I-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
their	NN	O	O
expression	NN	O	O
is	NN	O	O
influenced	NN	O	O
by	NN	O	O
exposure	NN	O	O
to	NN	O	O
the	NN	O	O
hormone	NN	O	O
.	NN	O	O

Estrogen	NN	O	B-protein
receptors	NN	O	I-protein
influence	NN	O	O
non-genomic	NN	O	O
events	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
rapid	NN	O	O
in	NN	O	O
onset	NN	O	O
and	NN	O	O
genomic	NN	O	O
events	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
longer	NN	O	O
acting	NN	O	O
responses	NN	O	O
.	NN	O	O

Estrogens	NN	O	O
affect	NN	O	O
vascular	NN	O	O
tone	NN	O	O
indirectly	NN	O	O
by	NN	O	O
modulating	NN	O	O
release	NN	O	O
of	NN	O	O
endothelium-derived	NN	O	B-protein
vasoactive	NN	O	I-protein
factors	NN	O	I-protein
and	NN	O	O
directly	NN	O	O
by	NN	O	O
modulating	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
in	NN	O	O
vascular	NN	O	B-cell_type
smooth	NN	O	I-cell_type
muscle	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Estrogens	NN	O	O
indirectly	NN	O	O
affect	NN	O	O
thrombotic	NN	O	O
events	NN	O	O
and	NN	O	O
inflammation	NN	O	O
by	NN	O	O
altering	NN	O	O
platelet	NN	O	B-cell_type
aggregation	NN	O	O
and	NN	O	O
leukocyte	NN	O	B-cell_type
adherence	NN	O	O
and	NN	O	O
migration	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Estrogens	NN	O	O
also	NN	O	O
influence	NN	O	O
production	NN	O	O
of	NN	O	O
mitogens	NN	O	O
which	NN	O	O
,	NN	O	O
when	NN	O	O
released	NN	O	O
at	NN	O	O
sites	NN	O	O
of	NN	O	O
vascular	NN	O	O
injury	NN	O	O
,	NN	O	O
affect	NN	O	O
vascular	NN	O	O
remodeling	NN	O	O
.	NN	O	O

Although	NN	O	O
estrogens	NN	O	O
initiate	NN	O	O
vascular	NN	O	O
responses	NN	O	O
,	NN	O	O
genomic	NN	O	O
sex	NN	O	O
may	NN	O	O
influence	NN	O	O
and/or	NN	O	O
limit	NN	O	O
expression	NN	O	O
of	NN	O	O
estrogen	NN	O	O
receptors	NN	O	O
and	NN	O	O
therefore	NN	O	O
actions	NN	O	O
of	NN	O	O
sex	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
throughout	NN	O	O
the	NN	O	O
vasculature	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
a	NN	O	O
functional	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
in	NN	O	O
all-trans-retinoic	NN	O	O
acid-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
M2-type	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
blast	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Different	NN	O	O
types	NN	O	O
of	NN	O	O
acute	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
blast	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
all-trans-retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
and	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
VD	NN	O	O
)	NN	O	O
.	NN	O	O

M0/M1	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
not	NN	O	O
sensitive	NN	O	O
to	NN	O	O
differentiating	NN	O	O
agents	NN	O	O
,	NN	O	O
whereas	NN	O	O
M3	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
induced	NN	O	O
to	NN	O	O
undergo	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
after	NN	O	O
ATRA	NN	O	O
treatment	NN	O	O
but	NN	O	O
are	NN	O	O
not	NN	O	O
sensitive	NN	O	O
to	NN	O	O
VD	NN	O	O
.	NN	O	O

M2	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
blast	NN	O	I-cell_line
cells	NN	O	I-cell_line
behave	NN	O	O
differently	NN	O	O
because	NN	O	O
they	NN	O	O
undergo	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
with	NN	O	O
both	NN	O	O
the	NN	O	O
differentiation	NN	O	O
inducers	NN	O	O
.	NN	O	O

To	NN	O	O
gain	NN	O	O
some	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
maturation	NN	O	O
of	NN	O	O
M2-type	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
ATRA	NN	O	O
and	NN	O	O
VD	NN	O	O
in	NN	O	O
spontaneous	NN	O	O
M2	NN	O	B-cell_line
blast	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
Kasumi-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
an	NN	O	O
acute	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
M2-type	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
ATRA	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
VD	NN	O	O
efficiently	NN	O	O
increases	NN	O	O
the	NN	O	O
nuclear	NN	O	O
abundance	NN	O	O
of	NN	O	O
VD	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
promotes	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

VDR	NN	O	B-protein
is	NN	O	O
functionally	NN	O	O
active	NN	O	O
in	NN	O	O
ATRA-treated	NN	O	O
Kasumi-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
because	NN	O	O
it	NN	O	O
efficiently	NN	O	O
heterodimerizes	NN	O	O
with	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
DR3-type	NN	O	B-DNA
vitamin	NN	O	I-DNA
D-responsive	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
activates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
vitamin	NN	O	B-DNA
D-responsive	NN	O	I-DNA
element-regulated	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
VD-responsive	NN	O	B-DNA
genes	NN	O	I-DNA
are	NN	O	O
induced	NN	O	O
by	NN	O	O
ATRA	NN	O	O
treatment	NN	O	O
of	NN	O	O
Kasumi-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
genetic	NN	O	O
program	NN	O	O
underlying	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
is	NN	O	O
activated	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
ATRA	NN	O	O
increases	NN	O	O
the	NN	O	O
nuclear	NN	O	O
abundance	NN	O	O
of	NN	O	O
a	NN	O	O
functional	NN	O	O
VDR	NN	O	B-protein
is	NN	O	O
still	NN	O	O
unknown	NN	O	O
,	NN	O	O
but	NN	O	O
our	NN	O	O
data	NN	O	O
clearly	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
M2	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
cell	NN	O	I-cell_line
context	NN	O	O
is	NN	O	O
only	NN	O	O
permissive	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
and	NN	O	O
calcineurin	NN	O	B-protein
synergize	NN	O	O
to	NN	O	O
activate	NN	O	O
IkappaB	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
of	NN	O	O
kappaB	NN	O	O
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
ubiquitous	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
is	NN	O	O
key	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
and	NN	O	O
inflammation	NN	O	O
.	NN	O	O

T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
cross-linking	NN	O	O
is	NN	O	O
in	NN	O	O
part	NN	O	O
required	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
.	NN	O	O

By	NN	O	O
using	NN	O	O
Jurkat	NN	O	B-cell_line
and	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
simultaneous	NN	O	O
activation	NN	O	O
of	NN	O	O
two	NN	O	O
second	NN	O	O
messengers	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
-initiated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
and	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IkappaBalpha	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
IKK	NN	O	I-protein
)	NN	O	I-protein
complex	NN	O	I-protein
but	NN	O	O
not	NN	O	O
of	NN	O	O
another	NN	O	O
putative	NN	O	B-protein
IkappaBalpha	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
p90	NN	O	B-protein
(	NN	O	I-protein
rsk	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
IKK	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
p90	NN	O	B-protein
(	NN	O	I-protein
rsk	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
in	NN	O	O
vivo	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
co-activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
and	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

Each	NN	O	O
second	NN	O	O
messenger	NN	O	O
is	NN	O	O
necessary	NN	O	O
,	NN	O	O
as	NN	O	O
inhibition	NN	O	O
of	NN	O	O
either	NN	O	O
one	NN	O	O
reverses	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IKK	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
IkappaBalpha	NN	O	B-protein
phosphorylation	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
dominant	NN	O	O
negative	NN	O	O
forms	NN	O	O
of	NN	O	O
IKKalpha	NN	O	B-protein
and	NN	O	I-protein
-beta	NN	O	I-protein
demonstrates	NN	O	O
that	NN	O	O
only	NN	O	O
IKKbeta	NN	O	B-protein
is	NN	O	O
the	NN	O	O
target	NN	O	O
for	NN	O	O
PKC	NN	O	B-protein
and	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
within	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
/CD3	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
both	NN	O	O
PKC	NN	O	B-protein
and	NN	O	O
calcineurin	NN	O	B-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
effective	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IKK	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Diminished	NN	O	O
responses	NN	O	O
to	NN	O	O
IL-13	NN	O	B-protein
by	NN	O	O
human	NN	O	O
monocytes	NN	O	O
differentiated	NN	O	O
in	NN	O	O
vitro	NN	O	O
:	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
IL-13Ralpha1	NN	O	B-protein
chain	NN	O	I-protein
and	NN	O	O
STAT6	NN	O	B-protein
.	NN	O	O

The	NN	O	O
primary	NN	O	O
IL-13	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
on	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
is	NN	O	O
believed	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
heterodimer	NN	O	O
comprised	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
and	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-protein
gamma	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	B-protein
gamma	NN	O	I-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	I-protein
-like	NN	O	I-protein
molecule	NN	O	I-protein
,	NN	O	O
IL-13R	NN	O	B-protein
alpha1	NN	O	I-protein
.	NN	O	O

mRNA	NN	O	B-RNA
levels	NN	O	O
for	NN	O	O
IL-13R	NN	O	B-protein
alpha1	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
were	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
in	NN	O	O
in	NN	O	B-cell_line
vitro	NN	O	I-cell_line
monocyte-derived	NN	O	I-cell_line
macrophages	NN	O	I-cell_line
(	NN	O	O
MDMac	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
and	NN	O	O
with	NN	O	O
increasing	NN	O	O
time	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
in	NN	O	O
culture	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
IL-13	NN	O	B-protein
regulation	NN	O	O
of	NN	O	O
lipopolysaccharide-induced	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
Daudi	NN	O	B-cell_line
and	NN	O	O
THP-1	NN	O	B-cell_line
that	NN	O	O
differentially	NN	O	O
express	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
IL-13R	NN	O	B-protein
alpha1	NN	O	I-protein
showed	NN	O	O
that	NN	O	O
IL-13	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
STAT6	NN	O	B-protein
in	NN	O	O
IL-13R	NN	O	B-cell_line
alpha1-positive	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
gamma	NN	O	B-cell_line
(	NN	O	I-cell_line
c	NN	O	I-cell_line
)	NN	O	I-cell_line
-positive	NN	O	I-cell_line
,	NN	O	I-cell_line
IL-13R	NN	O	I-cell_line
alpha1-negative	NN	O	I-cell_line
Daudi	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

IL-13	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
was	NN	O	O
reduced	NN	O	O
in	NN	O	O
MDMac	NN	O	B-cell_line
which	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
diminished	NN	O	O
IL-13	NN	O	B-protein
-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
CD23	NN	O	B-protein
and	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
with	NN	O	O
reduced	NN	O	O
IL-13R	NN	O	B-protein
alpha1	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
low	NN	O	O
nuclear	NN	O	O
STAT6	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
some	NN	O	O
IL-13	NN	O	B-protein
-induced	NN	O	O
responses	NN	O	O
were	NN	O	O
unaltered	NN	O	O
in	NN	O	O
magnitude	NN	O	O
in	NN	O	O
MDMac	NN	O	B-cell_line
.	NN	O	O

In	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
functional	NN	O	O
IL-13R	NN	O	B-protein
alpha1	NN	O	I-protein
and	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
IL-13	NN	O	B-protein
must	NN	O	O
signal	NN	O	O
through	NN	O	O
an	NN	O	O
alternative	NN	O	O
receptor	NN	O	B-protein
complex	NN	O	I-protein
on	NN	O	O
MDMac	NN	O	B-cell_line
.	NN	O	O

Experiments	NN	O	O
with	NN	O	O
a	NN	O	O
blocking	NN	O	O
antibody	NN	O	O
to	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
showed	NN	O	O
that	NN	O	O
this	NN	O	O
chain	NN	O	O
remains	NN	O	O
an	NN	O	O
essential	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
IL-13	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
on	NN	O	O
MDMac	NN	O	B-cell_line
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
p53	NN	O	B-protein
paradox	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
tumor	NN	O	O
progression	NN	O	O
.	NN	O	O

Recent	NN	O	O
evidence	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
p53	NN	O	B-protein
molecule	NN	O	I-protein
appears	NN	O	O
in	NN	O	O
two	NN	O	O
different	NN	O	O
forms	NN	O	O
:	NN	O	O
the	NN	O	O
mutant	NN	O	B-protein
p53	NN	O	I-protein
that	NN	O	O
stimulates	NN	O	O
tumor	NN	O	O
progression	NN	O	O
,	NN	O	O
and	NN	O	O
wild	NN	O	B-protein
type	NN	O	I-protein
p53	NN	O	I-protein
that	NN	O	O
inhibits	NN	O	O
tumor	NN	O	O
progression	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
established	NN	O	O
that	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
can	NN	O	O
activate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
wild	NN	O	B-protein
type	NN	O	I-protein
p53	NN	O	I-protein
in	NN	O	O
concert	NN	O	O
with	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Both	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
are	NN	O	O
also	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
pathway	NN	O	O
that	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	I-protein
MHC	NN	O	I-protein
)	NN	O	I-protein
class	NN	O	I-protein
I	NN	O	I-protein
molecules	NN	O	I-protein
and	NN	O	O
,	NN	O	O
hence	NN	O	O
,	NN	O	O
antigen	NN	O	O
presentation	NN	O	O
to	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
we	NN	O	O
shall	NN	O	O
advance	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
:	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
TNF-alpha	NN	O	B-protein
indirectly	NN	O	O
controls	NN	O	O
immune	NN	O	O
surveillance	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
TNF-alpha	NN	O	B-protein
controls	NN	O	O
DNA	NN	O	O
repair	NN	O	O
and	NN	O	O
tumor	NN	O	O
suppression	NN	O	O
through	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
wild	NN	O	B-protein
type	NN	O	I-protein
p53	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
elevated	NN	O	O
TNF-alpha	NN	O	B-protein
is	NN	O	O
primarily	NN	O	O
responsible	NN	O	O
for	NN	O	O
promoting	NN	O	O
tumor	NN	O	O
progression	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
promoter	NN	O	B-DNA
and	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
sequences	NN	O	I-DNA
controlling	NN	O	O
human	NN	O	O
B29	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

The	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	B-DNA
B29	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
B29	NN	O	B-protein
,	NN	O	O
Ig	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
CD79b	NN	O	B-protein
)	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
Ig-mediated	NN	O	O
B-cell	NN	O	O
activation	NN	O	O
via	NN	O	O
the	NN	O	O
B-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	O
BCR	NN	O	B-protein
)	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
and	NN	O	I-cell_type
murine	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
this	NN	O	O
pivotal	NN	O	O
gene	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
genomic	NN	O	I-DNA
DNA	NN	O	I-DNA
sequence	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
B29	NN	O	B-protein
ATG	NN	O	B-DNA
start	NN	O	I-DNA
codon	NN	O	I-DNA
for	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
B29	NN	O	B-DNA
gene	NN	O	I-DNA
lacks	NN	O	O
either	NN	O	O
a	NN	O	O
TATA	NN	O	B-DNA
or	NN	O	I-DNA
a	NN	O	I-DNA
CAAT	NN	O	I-DNA
box	NN	O	I-DNA
and	NN	O	O
transcription	NN	O	O
is	NN	O	O
initiated	NN	O	O
at	NN	O	O
multiple	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
minimal	NN	O	O
promoter	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
B29	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
contained	NN	O	O
within	NN	O	O
a	NN	O	O
193-bp	NN	O	B-DNA
region	NN	O	I-DNA
5	NN	O	O
'	NN	O	O
of	NN	O	O
these	NN	O	O
multiple	NN	O	B-DNA
start	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
minimal	NN	O	B-DNA
promoter	NN	O	I-DNA
exhibits	NN	O	O
B-cell-specific	NN	O	O
activity	NN	O	O
and	NN	O	O
contains	NN	O	O
SP1	NN	O	B-DNA
,	NN	O	I-DNA
ETS	NN	O	I-DNA
,	NN	O	I-DNA
OCT	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
IKAROS/LYF-1	NN	O	I-DNA
transcription	NN	O	I-DNA
factor	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

All	NN	O	O
these	NN	O	O
motifs	NN	O	O
are	NN	O	O
strikingly	NN	O	O
conserved	NN	O	O
in	NN	O	O
sequence	NN	O	O
and	NN	O	O
placement	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
previously	NN	O	O
characterized	NN	O	O
murine	NN	O	B-DNA
B29	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Additional	NN	O	O
upstream	NN	O	O
gene	NN	O	O
segments	NN	O	O
dramatically	NN	O	O
affected	NN	O	O
B29	NN	O	B-protein
minimal	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

A	NN	O	O
newly	NN	O	O
identified	NN	O	O
motif	NN	O	O
called	NN	O	O
the	NN	O	O
B29	NN	O	B-DNA
conserved	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
BCS	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
found	NN	O	O
upstream	NN	O	O
of	NN	O	O
both	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
murine	NN	O	I-DNA
B29	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
appears	NN	O	O
to	NN	O	O
stimulate	NN	O	O
B29	NN	O	B-protein
transcription	NN	O	O
through	NN	O	O
a	NN	O	O
novel	NN	O	O
mechanism	NN	O	O
.	NN	O	O

A	NN	O	O
single	NN	O	O
BCS	NN	O	B-DNA
had	NN	O	O
little	NN	O	O
effect	NN	O	O
either	NN	O	O
on	NN	O	O
the	NN	O	O
minimal	NN	O	O
B29	NN	O	B-DNA
promoter	NN	O	I-DNA
or	NN	O	O
on	NN	O	O
a	NN	O	O
heterologous	NN	O	O
promoter	NN	O	O
.	NN	O	O

Instead	NN	O	O
,	NN	O	O
the	NN	O	O
BCS	NN	O	B-DNA
stimulated	NN	O	O
transcription	NN	O	O
by	NN	O	O
counteracting	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
negative	NN	O	I-DNA
regulatory	NN	O	I-DNA
DNA	NN	O	I-DNA
sequences	NN	O	I-DNA
that	NN	O	O
block	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
B29	NN	O	B-protein
minimal	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
its	NN	O	O
absence	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
B29	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
the	NN	O	O
complex	NN	O	O
interplay	NN	O	O
of	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
immunosuppressive	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
bovine	NN	O	O
colostrum	NN	O	O
blocks	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
gene	NN	O	I-DNA
enhancer	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
factor	NN	O	O
in	NN	O	O
bovine	NN	O	O
colostrum	NN	O	O
(	NN	O	O
colostrum	NN	O	B-protein
inhibitory	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
CIF	NN	O	B-protein
)	NN	O	O
inhibits	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL2	NN	O	B-protein
)	NN	O	O
production	NN	O	O
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
helper	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
blocking	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
IL2	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
CIF	NN	O	B-protein
blocks	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
IL2	NN	O	B-protein
transcription	NN	O	O
,	NN	O	O
we	NN	O	O
introduced	NN	O	O
reporter	NN	O	O
plasmids	NN	O	O
into	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
by	NN	O	O
transient	NN	O	O
transfection	NN	O	O
.	NN	O	O

These	NN	O	O
contained	NN	O	O
the	NN	O	O
luciferase	NN	O	B-DNA
gene	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL2	NN	O	I-DNA
upstream	NN	O	I-DNA
enhancer	NN	O	I-DNA
region	NN	O	O
(	NN	O	O
segments	NN	O	O
-326	NN	O	O
to	NN	O	O
+45	NN	O	O
)	NN	O	O
or	NN	O	O
three	NN	O	O
repeats	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
element	NN	O	I-DNA
contained	NN	O	O
within	NN	O	O
it	NN	O	O
(	NN	O	O
segments	NN	O	O
-255	NN	O	O
to	NN	O	O
-285	NN	O	O
)	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
luciferase	NN	O	B-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
plus	NN	O	O
a	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
.	NN	O	O

CIF	NN	O	B-protein
inhibited	NN	O	O
induction	NN	O	O
of	NN	O	O
either	NN	O	O
construct	NN	O	O
as	NN	O	O
did	NN	O	O
cyclosporine	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
block	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
element	NN	O	I-protein
.	NN	O	O

CIF	NN	O	B-protein
failed	NN	O	O
to	NN	O	O
inhibit	NN	O	O
several	NN	O	O
other	NN	O	O
enhancer	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
NFAT-controlled	NN	O	B-DNA
luciferase	NN	O	I-DNA
gene	NN	O	I-DNA
system	NN	O	I-DNA
distinguishes	NN	O	O
CIF	NN	O	B-protein
from	NN	O	O
other	NN	O	O
T	NN	O	O
cell	NN	O	O
inhibitory	NN	O	O
activities	NN	O	O
present	NN	O	O
in	NN	O	O
colostrum	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
TGF	NN	O	B-protein
beta	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
TGF	NN	O	B-protein
beta	NN	O	I-protein
2	NN	O	I-protein
and	NN	O	O
the	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Stably	NN	O	O
transfected	NN	O	O
Jurkat	NN	O	O
cells	NN	O	O
behaved	NN	O	O
similarly	NN	O	O
to	NN	O	O
the	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
ones	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
inhibition	NN	O	O
by	NN	O	O
CIF	NN	O	B-protein
and	NN	O	O
cyclosporine	NN	O	O
.	NN	O	O

The	NN	O	O
NFAT	NN	O	B-protein
-luc	NN	O	O
assay	NN	O	O
is	NN	O	O
a	NN	O	O
useful	NN	O	O
technique	NN	O	O
for	NN	O	O
the	NN	O	O
rapid	NN	O	O
,	NN	O	O
sensitive	NN	O	O
measurement	NN	O	O
of	NN	O	O
CIF	NN	O	B-protein
or	NN	O	O
other	NN	O	O
immunosuppressants	NN	O	O
with	NN	O	O
a	NN	O	O
similar	NN	O	O
mode	NN	O	O
of	NN	O	O
action	NN	O	O
.	NN	O	O

-DOCSTART-	O

Heat	NN	O	O
shock	NN	O	O
induces	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
chronically	NN	O	O
infected	NN	O	O
promyelocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
OM10.1	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
long	NN	O	O
period	NN	O	O
of	NN	O	O
clinical	NN	O	O
latency	NN	O	O
before	NN	O	O
development	NN	O	O
of	NN	O	O
symptoms	NN	O	O
is	NN	O	O
characteristic	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
infection	NN	O	O
.	NN	O	O

OM10.1	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
promyelocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
latently	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
developed	NN	O	O
as	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
studying	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
viral	NN	O	O
latency	NN	O	O
and	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
virus	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
this	NN	O	O
latently	NN	O	O
infected	NN	O	O
cell	NN	O	O
line	NN	O	O
with	NN	O	O
heat	NN	O	O
shock	NN	O	O
at	NN	O	O
42	NN	O	O
degrees	NN	O	O
C	NN	O	O
for	NN	O	O
2	NN	O	O
h	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
without	NN	O	O
addition	NN	O	O
of	NN	O	O
any	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
activation	NN	O	O
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
using	NN	O	O
anti-TNF-alpha	NN	O	B-protein
antibody	NN	O	I-protein
and	NN	O	O
various	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-protein
level	NN	O	O
in	NN	O	O
culture	NN	O	O
supernatants	NN	O	O
was	NN	O	O
below	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
an	NN	O	O
ELISA	NN	O	O
assay	NN	O	O
system	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
anti-	NN	O	O
TNF-alpha	NN	O	B-protein
antibody	NN	O	O
in	NN	O	O
culture	NN	O	O
medium	NN	O	O
could	NN	O	O
partially	NN	O	O
suppress	NN	O	O
the	NN	O	O
heat	NN	O	O
shock	NN	O	O
induced	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
.	NN	O	O

Staurosporine	NN	O	O
(	NN	O	O
PKC	NN	O	O
inhibitor	NN	O	O
)	NN	O	O
,	NN	O	O
pentoxifylline	NN	O	O
(	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
inhibitor	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
Ro5-3335	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
Tat	NN	O	B-protein
inhibitor	NN	O	O
)	NN	O	O
also	NN	O	O
inhibited	NN	O	O
significantly	NN	O	O
the	NN	O	O
heat	NN	O	O
shock	NN	O	O
induced	NN	O	O
virus	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
staurosporine	NN	O	O
achieved	NN	O	O
approximately	NN	O	O
90	NN	O	O
%	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
antigen	NN	O	O
expression	NN	O	O
in	NN	O	O
heat	NN	O	B-cell_line
shock-treated	NN	O	I-cell_line
OM10.1	NN	O	I-cell_line
at	NN	O	O
a	NN	O	O
non-toxic	NN	O	O
concentration	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
activation	NN	O	O
with	NN	O	O
heat	NN	O	O
shock	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
fully	NN	O	O
elucidated	NN	O	O
yet	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
presumed	NN	O	O
PKC	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
HIV-1	NN	O	O
activation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
present	NN	O	O
observations	NN	O	O
will	NN	O	O
provide	NN	O	O
a	NN	O	O
further	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
infections	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
CDC42	NN	O	B-protein
effector	NN	O	I-protein
N-WASP	NN	O	B-protein
by	NN	O	O
the	NN	O	O
Shigella	NN	O	B-protein
flexneri	NN	O	I-protein
IcsA	NN	O	I-protein
protein	NN	O	I-protein
promotes	NN	O	O
actin	NN	O	B-protein
nucleation	NN	O	O
by	NN	O	O
Arp2/3	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
bacterial	NN	O	O
actin	NN	O	B-protein
-based	NN	O	O
motility	NN	O	O
.	NN	O	O

To	NN	O	O
propel	NN	O	O
itself	NN	O	O
in	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
pathogen	NN	O	O
Shigella	NN	O	O
flexneri	NN	O	O
subverts	NN	O	O
the	NN	O	O
Cdc42	NN	O	B-protein
-controlled	NN	O	O
machinery	NN	O	O
responsible	NN	O	O
for	NN	O	O
actin	NN	O	B-protein
assembly	NN	O	O
during	NN	O	O
filopodia	NN	O	O
formation	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
bacterial	NN	O	O
motility	NN	O	O
assays	NN	O	O
in	NN	O	O
platelet	NN	O	O
extracts	NN	O	O
with	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
expressing	NN	O	O
the	NN	O	O
Shigella	NN	O	B-protein
IcsA	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
analysis	NN	O	O
of	NN	O	O
reconstituted	NN	O	O
systems	NN	O	O
from	NN	O	O
purified	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
we	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
bacterial	NN	O	B-protein
protein	NN	O	I-protein
IcsA	NN	O	I-protein
binds	NN	O	O
N-WASP	NN	O	B-protein
and	NN	O	O
activates	NN	O	O
it	NN	O	O
in	NN	O	O
a	NN	O	O
Cdc42	NN	O	B-protein
-like	NN	O	O
fashion	NN	O	O
.	NN	O	O

Dramatic	NN	O	O
stimulation	NN	O	O
of	NN	O	O
actin	NN	O	B-protein
assembly	NN	O	O
is	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
ternary	NN	O	O
IcsA-N-WASP-Arp2/3	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
which	NN	O	O
nucleates	NN	O	O
actin	NN	O	B-protein
polymerization	NN	O	O
.	NN	O	O

The	NN	O	O
Arp2/3	NN	O	B-protein
complex	NN	O	I-protein
is	NN	O	O
essential	NN	O	O
in	NN	O	O
initiation	NN	O	O
of	NN	O	O
actin	NN	O	B-protein
assembly	NN	O	O
and	NN	O	O
Shigella	NN	O	O
movement	NN	O	O
,	NN	O	O
as	NN	O	O
previously	NN	O	O
observed	NN	O	O
for	NN	O	O
Listeria	NN	O	O
monocytogenes	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
N-WASP	NN	O	B-protein
by	NN	O	O
IcsA	NN	O	B-protein
unmasks	NN	O	O
two	NN	O	O
domains	NN	O	O
acting	NN	O	O
together	NN	O	O
in	NN	O	O
insertional	NN	O	O
actin	NN	O	B-protein
polymerization	NN	O	O
.	NN	O	O

The	NN	O	O
isolated	NN	O	O
COOH-terminal	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
N-WASP	NN	O	B-protein
containing	NN	O	O
a	NN	O	O
verprolin-homology	NN	O	B-protein
region	NN	O	I-protein
,	NN	O	O
a	NN	O	O
cofilin-homology	NN	O	O
sequence	NN	O	O
,	NN	O	O
and	NN	O	O
an	NN	O	O
acidic	NN	O	B-protein
terminal	NN	O	I-protein
segment	NN	O	I-protein
(	NN	O	O
VCA	NN	O	B-protein
)	NN	O	O
interacts	NN	O	O
with	NN	O	O
G-actin	NN	O	B-protein
in	NN	O	O
a	NN	O	O
unique	NN	O	O
profilin-like	NN	O	O
functional	NN	O	O
fashion	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
when	NN	O	O
N-WASP	NN	O	B-protein
is	NN	O	O
activated	NN	O	O
,	NN	O	O
its	NN	O	O
COOH-terminal	NN	O	B-protein
domain	NN	O	I-protein
feeds	NN	O	O
barbed	NN	O	O
end	NN	O	O
growth	NN	O	O
of	NN	O	O
filaments	NN	O	O
and	NN	O	O
lowers	NN	O	O
the	NN	O	O
critical	NN	O	O
concentration	NN	O	O
at	NN	O	O
the	NN	O	O
bacterial	NN	O	O
surface	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
the	NN	O	O
NH	NN	O	B-protein
(	NN	O	I-protein
2	NN	O	I-protein
)	NN	O	I-protein
-terminal	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
N-WASP	NN	O	B-protein
interacts	NN	O	O
with	NN	O	O
F-actin	NN	O	B-protein
,	NN	O	O
mediating	NN	O	O
the	NN	O	O
attachment	NN	O	O
of	NN	O	O
the	NN	O	O
actin	NN	O	B-protein
tail	NN	O	I-protein
to	NN	O	O
the	NN	O	O
bacterium	NN	O	O
surface	NN	O	O
.	NN	O	O

VASP	NN	O	O
is	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
Shigella	NN	O	O
movement	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
profilin	NN	O	B-protein
does	NN	O	O
not	NN	O	O
require	NN	O	O
its	NN	O	O
binding	NN	O	O
to	NN	O	O
proline-rich	NN	O	B-protein
regions	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

c-Maf	NN	O	B-protein
induces	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
bipotent	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
drive	NN	O	O
colony-forming	NN	O	B-cell_line
unit	NN	O	I-cell_line
granulocyte-macrophage	NN	O	I-cell_line
(	NN	O	I-cell_line
CFU-GM	NN	O	I-cell_line
)	NN	O	I-cell_line
myeloid	NN	O	I-cell_line
progenitors	NN	O	I-cell_line
to	NN	O	O
differentiate	NN	O	O
into	NN	O	O
cells	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
granulocytic	NN	O	B-cell_type
or	NN	O	I-cell_type
monocytic	NN	O	I-cell_type
lineage	NN	O	I-cell_type
are	NN	O	O
not	NN	O	O
fully	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
c-Maf	NN	O	B-protein
and	NN	O	O
c-Myb	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
physically	NN	O	O
interact	NN	O	O
in	NN	O	O
myeloid	NN	O	O
cells	NN	O	O
to	NN	O	O
form	NN	O	O
inhibitory	NN	O	O
complexes	NN	O	O
that	NN	O	O
hinder	NN	O	O
transactivation	NN	O	O
of	NN	O	O
c-Myb	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
through	NN	O	O
direct	NN	O	O
binding	NN	O	O
to	NN	O	O
Myb	NN	O	B-DNA
consensus	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
complexes	NN	O	O
arise	NN	O	O
in	NN	O	O
a	NN	O	O
developmentally	NN	O	O
regulated	NN	O	O
pattern	NN	O	O
,	NN	O	O
peaking	NN	O	O
at	NN	O	O
the	NN	O	O
promyelocyte	NN	O	B-cell_type
stage	NN	O	I-cell_type
,	NN	O	O
or	NN	O	O
in	NN	O	O
cell	NN	O	B-cell_line
model	NN	O	I-cell_line
systems	NN	O	I-cell_line
,	NN	O	O
appearing	NN	O	O
soon	NN	O	O
after	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
wished	NN	O	O
to	NN	O	O
determine	NN	O	O
if	NN	O	O
this	NN	O	O
developmentally	NN	O	O
related	NN	O	O
interaction	NN	O	O
is	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
or	NN	O	O
an	NN	O	O
intrinsic	NN	O	O
differentiating	NN	O	O
stimulus	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
elevated	NN	O	O
Myb	NN	O	B-protein
:	NN	O	I-protein
Maf	NN	O	I-protein
status	NN	O	O
seen	NN	O	O
in	NN	O	O
differentiating	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
be	NN	O	O
recapitulated	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
c-Maf	NN	O	B-protein
in	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
lines	NN	O	O
,	NN	O	O
we	NN	O	O
inducibly	NN	O	O
expressed	NN	O	O
the	NN	O	O
c-Maf	NN	O	B-DNA
cDNA	NN	O	I-DNA
in	NN	O	O
2	NN	O	O
bipotent	NN	O	O
human	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
c-Maf	NN	O	B-protein
protein	NN	O	I-protein
led	NN	O	O
to	NN	O	O
marked	NN	O	O
increases	NN	O	O
in	NN	O	O
Myb	NN	O	B-protein
:	NN	O	I-protein
Maf	NN	O	I-protein
complexes	NN	O	I-protein
and	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
followed	NN	O	O
by	NN	O	O
eventual	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
targets	NN	O	O
that	NN	O	O
could	NN	O	O
mediate	NN	O	O
these	NN	O	O
phenotypic	NN	O	O
changes	NN	O	O
indicated	NN	O	O
that	NN	O	O
c-Maf	NN	O	B-protein
likely	NN	O	O
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
development	NN	O	O
through	NN	O	O
dual	NN	O	O
mechanisms	NN	O	O
;	NN	O	O
inhibition	NN	O	O
of	NN	O	O
a	NN	O	O
select	NN	O	O
set	NN	O	O
of	NN	O	O
c-Myb	NN	O	B-protein
regulated	NN	O	O
targets	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
Bcl-2	NN	O	B-protein
and	NN	O	O
CD13/APN	NN	O	B-protein
,	NN	O	O
coupled	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
as	NN	O	O
yet	NN	O	O
undefined	NN	O	O
differentiation-promoting	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

AML1	NN	O	B-protein
(	NN	O	I-protein
CBFalpha2	NN	O	I-protein
)	NN	O	I-protein
cooperates	NN	O	O
with	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
activating	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
BSAP/PAX5	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
B	NN	O	B-DNA
cell-specific	NN	O	I-DNA
BLK	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

AML1	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
during	NN	O	O
hematopoiesis	NN	O	O
and	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
involving	NN	O	O
AML1	NN	O	B-protein
are	NN	O	O
commonly	NN	O	O
associated	NN	O	O
with	NN	O	O
different	NN	O	O
forms	NN	O	O
of	NN	O	O
leukemia	NN	O	O
,	NN	O	O
including	NN	O	O
pre-B	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
AML1	NN	O	B-protein
during	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
regulatory	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
B	NN	O	B-DNA
cell-specific	NN	O	I-DNA
genes	NN	O	I-DNA
for	NN	O	O
potential	NN	O	O
AML1-binding	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
putative	NN	O	O
AML1-binding	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoter	NN	O	O
of	NN	O	O
the	NN	O	O
B	NN	O	B-DNA
cell-specific	NN	O	I-DNA
tyrosine	NN	O	I-DNA
kinase	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	I-DNA
blk	NN	O	I-DNA
.	NN	O	O

Gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
and	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
AML1	NN	O	B-protein
binds	NN	O	O
specifically	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
in	NN	O	O
the	NN	O	O
blk	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
this	NN	O	O
binding	NN	O	O
site	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
blk	NN	O	B-DNA
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
binding	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
AML1	NN	O	B-DNA
runt	NN	O	I-DNA
DNA-binding	NN	O	I-DNA
domain	NN	O	I-DNA
physically	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
paired	NN	O	B-DNA
DNA-binding	NN	O	I-DNA
domain	NN	O	I-DNA
of	NN	O	O
BSAP	NN	O	B-protein
,	NN	O	O
a	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

BSAP	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
previously	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
blk	NN	O	B-DNA
gene	NN	O	O
.	NN	O	O

Physical	NN	O	O
interaction	NN	O	O
of	NN	O	O
AML1	NN	O	B-protein
with	NN	O	O
BSAP	NN	O	B-protein
correlates	NN	O	O
with	NN	O	O
functional	NN	O	O
cooperativity	NN	O	O
in	NN	O	O
transfection	NN	O	O
studies	NN	O	O
where	NN	O	O
AML1	NN	O	B-protein
and	NN	O	O
BSAP	NN	O	B-protein
synergistically	NN	O	O
activate	NN	O	O
blk	NN	O	B-DNA
promoter	NN	O	O
transcription	NN	O	O
by	NN	O	O
more	NN	O	O
than	NN	O	O
50-fold	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
physical	NN	O	O
and	NN	O	O
functional	NN	O	O
interactions	NN	O	O
between	NN	O	O
AML1	NN	O	B-protein
and	NN	O	O
BSAP	NN	O	B-protein
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
AML1	NN	O	B-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
factor	NN	O	O
for	NN	O	O
regulating	NN	O	O
a	NN	O	O
critical	NN	O	O
B	NN	O	B-DNA
cell-specific	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
blk	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
inhibition	NN	O	O
by	NN	O	O
interleukin-6	NN	O	B-protein
of	NN	O	O
the	NN	O	O
class	NN	O	B-protein
A	NN	O	I-protein
macrophage	NN	O	I-protein
scavenger	NN	O	I-protein
receptor	NN	O	I-protein
in	NN	O	O
macrophages	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
THP-1	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-protein
A	NN	O	I-protein
macrophage	NN	O	I-protein
scavenger	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
MSR	NN	O	I-protein
)	NN	O	I-protein
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
uptake	NN	O	O
of	NN	O	O
modified	NN	O	B-protein
low	NN	O	I-protein
density	NN	O	I-protein
lipoproteins	NN	O	I-protein
(	NN	O	I-protein
LDL	NN	O	I-protein
)	NN	O	I-protein
by	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
transformation	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
into	NN	O	O
lipid-laden	NN	O	B-cell_type
foam	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
characterize	NN	O	O
atherosclerosis	NN	O	O
.	NN	O	O

Many	NN	O	O
environmental	NN	O	O
factors	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
and	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
can	NN	O	O
exert	NN	O	O
regulatory	NN	O	O
effects	NN	O	O
on	NN	O	O
MSR	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
whereas	NN	O	O
intracellular	NN	O	O
accumulation	NN	O	O
of	NN	O	O
cholesterol	NN	O	O
itself	NN	O	O
does	NN	O	O
not	NN	O	O
influence	NN	O	O
MSR	NN	O	B-protein
levels	NN	O	O
to	NN	O	O
any	NN	O	O
considerable	NN	O	O
extent	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
by	NN	O	O
using	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
model	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
whether	NN	O	O
stimulation	NN	O	O
with	NN	O	O
interleukin-6	NN	O	B-protein
(	NN	O	I-protein
IL-6	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
an	NN	O	O
immunoregulatory	NN	O	B-protein
,	NN	O	O
multipotential	NN	O	B-protein
cytokine	NN	O	I-protein
,	NN	O	O
modulates	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
activities	NN	O	O
of	NN	O	O
the	NN	O	O
MSR	NN	O	B-protein
in	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

When	NN	O	O
treated	NN	O	O
with	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
macrophages	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
peripheral	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
phorbol	NN	O	B-cell_line
12-myristate	NN	O	I-cell_line
13-acetate	NN	O	I-cell_line
(	NN	O	I-cell_line
PMA	NN	O	I-cell_line
)	NN	O	I-cell_line
-differentiated	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
showed	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
uptake	NN	O	O
and/or	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
MSR	NN	O	B-protein
ligand	NN	O	O
,	NN	O	O
acetylated	NN	O	B-protein
LDL	NN	O	I-protein
.	NN	O	O

This	NN	O	O
effect	NN	O	O
was	NN	O	O
paralleled	NN	O	O
by	NN	O	O
a	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
MSR	NN	O	B-protein
protein	NN	O	O
and	NN	O	O
mRNA	NN	O	B-RNA
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
MSR	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
transfected	NN	O	O
with	NN	O	O
an	NN	O	O
MSR	NN	O	B-DNA
promoter-reporter	NN	O	I-DNA
gene	NN	O	I-DNA
construct	NN	O	I-DNA
demonstrated	NN	O	O
decreased	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
MSR	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
IL-6	NN	O	B-protein
-treated	NN	O	O
THP-1	NN	O	B-cell_line
macrophages	NN	O	B-cell_type
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
gel	NN	O	O
shift	NN	O	O
assay	NN	O	O
also	NN	O	O
showed	NN	O	O
a	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
to	NN	O	O
the	NN	O	O
MSR	NN	O	B-DNA
promoter	NN	O	I-DNA
AP-1/ets	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
IL-6	NN	O	B-protein
-treated	NN	O	O
cells	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
exposure	NN	O	O
to	NN	O	O
IL-6	NN	O	B-protein
may	NN	O	O
inhibit	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-protein
A	NN	O	I-protein
MSR	NN	O	I-protein
in	NN	O	O
differentiated	NN	O	O
macrophages	NN	O	B-cell_type
at	NN	O	O
transcriptional	NN	O	O
levels	NN	O	O
.	NN	O	O

This	NN	O	O
result	NN	O	O
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
cytokine	NN	O	B-protein
may	NN	O	O
modulate	NN	O	O
foam	NN	O	O
cell	NN	O	O
formation	NN	O	O
during	NN	O	O
atherogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Retinoblastoma	NN	O	O
protein	NN	O	O
expression	NN	O	O
leads	NN	O	O
to	NN	O	O
reduced	NN	O	O
Oct-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
enhances	NN	O	O
interleukin-8	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Tumor	NN	O	O
cell	NN	O	O
lines	NN	O	O
with	NN	O	O
a	NN	O	O
defective	NN	O	O
retinoblastoma	NN	O	B-DNA
gene	NN	O	I-DNA
are	NN	O	O
unable	NN	O	O
to	NN	O	O
transcribe	NN	O	O
the	NN	O	O
HLA	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
treatment	NN	O	O
,	NN	O	O
and	NN	O	O
reconstitution	NN	O	O
of	NN	O	O
functional	NN	O	O
Rb	NN	O	B-protein
rescues	NN	O	O
IFN-gamma-induced	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
Rb	NN	O	B-protein
rescue	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
Rb	NN	O	B-protein
expression	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
for	NN	O	O
HLA-DRA	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
prototype	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Oct-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
POU	NN	O	B-protein
domain	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
repressor	NN	O	O
of	NN	O	O
HLA-DRA	NN	O	B-DNA
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
Rb-defective	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Rb	NN	O	B-protein
expression	NN	O	O
led	NN	O	O
to	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Oct-1	NN	O	B-protein
,	NN	O	O
thus	NN	O	O
relieving	NN	O	O
its	NN	O	O
repressive	NN	O	O
effect	NN	O	O
.	NN	O	O

Oct-1	NN	O	B-protein
has	NN	O	O
also	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
repress	NN	O	O
interleukin	NN	O	B-protein
8	NN	O	I-protein
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
Oct-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
Rb-transformed	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
interleukin	NN	O	B-protein
8	NN	O	I-protein
expression	NN	O	O
is	NN	O	O
higher	NN	O	O
in	NN	O	O
these	NN	O	O
cell	NN	O	O
lines	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cell	NN	O	O
activation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
bacterial	NN	O	B-protein
lipoproteins	NN	O	I-protein
through	NN	O	O
toll-like	NN	O	B-protein
receptor-2	NN	O	I-protein
.	NN	O	O

Apoptosis	NN	O	O
is	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
and	NN	O	O
resolution	NN	O	O
of	NN	O	O
inflammation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
bacterial	NN	O	O
pathogens	NN	O	O
.	NN	O	O

All	NN	O	O
bacterial	NN	O	O
pathogens	NN	O	O
produce	NN	O	O
lipoproteins	NN	O	B-protein
(	NN	O	I-protein
BLPs	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
which	NN	O	O
trigger	NN	O	O
the	NN	O	O
innate	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

BLPs	NN	O	B-protein
were	NN	O	O
found	NN	O	O
to	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
through	NN	O	O
human	NN	O	B-protein
Toll-like	NN	O	I-protein
receptor-2	NN	O	I-protein
(	NN	O	I-protein
hTLR2	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

BLPs	NN	O	B-protein
also	NN	O	O
initiated	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
an	NN	O	O
epithelial	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
transfected	NN	O	O
with	NN	O	O
hTLR2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
BLPs	NN	O	B-protein
stimulated	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
,	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
of	NN	O	O
multiple	NN	O	B-DNA
host	NN	O	I-DNA
defense	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
activated	NN	O	O
the	NN	O	O
respiratory	NN	O	O
burst	NN	O	O
through	NN	O	O
hTLR2	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
hTLR2	NN	O	B-protein
is	NN	O	O
a	NN	O	O
molecular	NN	O	O
link	NN	O	O
between	NN	O	O
microbial	NN	O	O
products	NN	O	O
,	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
and	NN	O	O
host	NN	O	O
defense	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

-DOCSTART-	O

Possible	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
different	NN	O	O
glucocorticoid	NN	O	O
hormone	NN	O	O
compounds	NN	O	O
.	NN	O	O

Different	NN	O	O
glucocorticoid	NN	O	O
hormones	NN	O	O
(	NN	O	O
GCH	NN	O	O
)	NN	O	O
show	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
intensity	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
their	NN	O	O
immunomodulating	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
GCH	NN	O	O
is	NN	O	O
under	NN	O	O
investigation	NN	O	O
,	NN	O	O
but	NN	O	O
is	NN	O	O
has	NN	O	O
been	NN	O	O
noted	NN	O	O
that	NN	O	O
they	NN	O	O
exert	NN	O	O
immune	NN	O	O
activity	NN	O	O
via	NN	O	O
the	NN	O	O
genomic	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
prednisone	NN	O	O
(	NN	O	O
PDN	NN	O	O
)	NN	O	O
,	NN	O	O
deflazacort	NN	O	O
(	NN	O	O
DFC	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
DXM	NN	O	O
)	NN	O	O
on	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
by	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
by	NN	O	O
activated	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
data	NN	O	O
obtained	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
three	NN	O	O
GCH	NN	O	O
molecules	NN	O	O
exert	NN	O	O
an	NN	O	O
immunosuppression	NN	O	O
on	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
a	NN	O	O
strong	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kB	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
;	NN	O	O
moreover	NN	O	O
,	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
not	NN	O	O
all	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
investigated	NN	O	O
were	NN	O	O
affected	NN	O	O
,	NN	O	O
and	NN	O	O
not	NN	O	O
with	NN	O	O
the	NN	O	O
same	NN	O	O
intensity	NN	O	O
,	NN	O	O
by	NN	O	O
the	NN	O	O
three	NN	O	O
GCH	NN	O	O
and	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
DXM	NN	O	O
inhibited	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kB	NN	O	B-protein
less	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
DFC	NN	O	O
and	NN	O	O
PDN	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
in	NN	O	O
agreement	NN	O	O
with	NN	O	O
the	NN	O	O
concept	NN	O	O
that	NN	O	O
different	NN	O	O
GCH	NN	O	O
compounds	NN	O	O
might	NN	O	O
differ	NN	O	O
in	NN	O	O
their	NN	O	O
binding	NN	O	O
and	NN	O	O
affinity	NN	O	O
properties	NN	O	O
,	NN	O	O
tissue-specific	NN	O	O
metabolism	NN	O	O
,	NN	O	O
and	NN	O	O
interaction	NN	O	O
with	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Distinctive	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
nodular	NN	O	O
lymphocyte	NN	O	O
predominance	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
BCL-6	NN	O	B-RNA
gene	NN	O	I-RNA
encoding	NN	O	O
a	NN	O	O
nuclear-located	NN	O	B-protein
Kruppel-type	NN	O	I-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
rearranged	NN	O	O
in	NN	O	O
about	NN	O	O
30	NN	O	O
%	NN	O	O
diffuse	NN	O	O
large	NN	O	O
B-cell	NN	O	B-cell_type
lymphomas	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
predominantly	NN	O	O
in	NN	O	O
normal	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
related	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
BCL-6	NN	O	B-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
differentiation	NN	O	O
of	NN	O	O
normal	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
its	NN	O	O
deregulated	NN	O	O
expression	NN	O	O
caused	NN	O	O
by	NN	O	O
rearrangements	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
lymphomagenesis	NN	O	O
.	NN	O	O

This	NN	O	O
prompted	NN	O	O
us	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
(	NN	O	O
HD	NN	O	O
)	NN	O	O
,	NN	O	O
focusing	NN	O	O
on	NN	O	O
the	NN	O	O
nodular	NN	O	O
lymphocyte	NN	O	O
predominance	NN	O	O
subtype	NN	O	O
(	NN	O	O
NLPHD	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
differs	NN	O	O
from	NN	O	O
classical	NN	O	O
HD	NN	O	O
by	NN	O	O
virtue	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
malignant	NN	O	B-cell_type
cell	NN	O	I-cell_type
population	NN	O	I-cell_type
(	NN	O	O
so-called	NN	O	O
L	NN	O	B-cell_type
&	NN	O	I-cell_type
H	NN	O	I-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
and	NN	O	O
its	NN	O	O
relationship	NN	O	O
with	NN	O	O
germinal	NN	O	O
centers	NN	O	O
.	NN	O	O

Forty-one	NN	O	O
HD	NN	O	O
samples	NN	O	O
(	NN	O	O
19	NN	O	O
NLPHD	NN	O	O
,	NN	O	O
12	NN	O	O
nodular	NN	O	O
sclerosis	NN	O	O
,	NN	O	O
and	NN	O	O
10	NN	O	O
mixed	NN	O	O
cellularity	NN	O	O
)	NN	O	O
were	NN	O	O
immunostained	NN	O	O
with	NN	O	O
the	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
PG-B6	NN	O	B-protein
and	NN	O	O
PG-B6p	NN	O	B-protein
that	NN	O	O
react	NN	O	O
with	NN	O	O
a	NN	O	O
fixative-sensitive	NN	O	O
and	NN	O	O
a	NN	O	O
formalin-resistant	NN	O	B-protein
epitope	NN	O	I-protein
on	NN	O	O
the	NN	O	O
aminoterminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Strong	NN	O	O
nuclear	NN	O	O
positivity	NN	O	O
for	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
tumor	NN	O	B-cell_type
(	NN	O	I-cell_type
L	NN	O	I-cell_type
&	NN	O	I-cell_type
H	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
all	NN	O	O
cases	NN	O	O
of	NN	O	O
NLPHD	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
BCL-6	NN	O	B-protein
was	NN	O	O
expressed	NN	O	O
only	NN	O	O
in	NN	O	O
a	NN	O	O
small	NN	O	O
percentage	NN	O	O
of	NN	O	O
Hodgkin	NN	O	B-cell_line
and	NN	O	I-cell_line
Reed-Sternberg	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
about	NN	O	O
30	NN	O	O
%	NN	O	O
of	NN	O	O
classical	NN	O	O
HD	NN	O	O
cases	NN	O	O
.	NN	O	O

Notably	NN	O	O
,	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
reactive	NN	O	B-cell_type
CD3+/CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
nearby	NN	O	O
to	NN	O	O
and	NN	O	O
rosetting	NN	O	O
around	NN	O	O
L	NN	O	B-cell_type
&	NN	O	I-cell_type
H	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
NLPHD	NN	O	O
were	NN	O	O
also	NN	O	O
strongly	NN	O	O
BCL-6	NN	O	B-protein
+	NN	O	O
,	NN	O	O
but	NN	O	O
lacked	NN	O	O
CD40	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
CD40L	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
.	NN	O	O

This	NN	O	O
staining	NN	O	O
pattern	NN	O	O
clearly	NN	O	O
differed	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
classical	NN	O	O
HD	NN	O	O
,	NN	O	O
whose	NN	O	O
cellular	NN	O	O
background	NN	O	O
was	NN	O	O
made	NN	O	O
up	NN	O	O
of	NN	O	O
CD3+/CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
showing	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
-/	NN	O	O
CD40L	NN	O	B-protein
+	NN	O	O
phenotype	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
further	NN	O	O
support	NN	O	O
the	NN	O	O
concept	NN	O	O
that	NN	O	O
NLPHD	NN	O	O
is	NN	O	O
an	NN	O	O
histogenetically	NN	O	O
distinct	NN	O	O
,	NN	O	O
B-cell-derived	NN	O	O
subtype	NN	O	O
of	NN	O	O
HD	NN	O	O
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
BCL-6	NN	O	B-protein
in	NN	O	O
its	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Selenium-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

The	NN	O	O
eukaryotic	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
various	NN	O	O
inflammatory	NN	O	B-DNA
genes	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
induced	NN	O	O
by	NN	O	O
prooxidants	NN	O	O
and	NN	O	O
several	NN	O	O
stimuli	NN	O	O
eliciting	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
lipopolysaccharide	NN	O	O
,	NN	O	O
UV	NN	O	O
irradiation	NN	O	O
and	NN	O	O
other	NN	O	O
mediators	NN	O	O
.	NN	O	O

Various	NN	O	O
antioxidants	NN	O	O
inhibit	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
these	NN	O	O
stimuli	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
selenium	NN	O	O
,	NN	O	O
an	NN	O	O
integral	NN	O	O
component	NN	O	O
of	NN	O	O
glutathione	NN	O	B-protein
peroxidase	NN	O	I-protein
(	NN	O	O
GPX	NN	O	B-protein
)	NN	O	O
,	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

In	NN	O	O
selenium-deprived	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
and	NN	O	O
ESb-L	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
,	NN	O	O
supplementation	NN	O	O
of	NN	O	O
selenium	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
substantial	NN	O	O
increase	NN	O	O
of	NN	O	O
GPX	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
revealed	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TNF	NN	O	B-protein
was	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
under	NN	O	O
these	NN	O	O
conditions	NN	O	O
.	NN	O	O

Likewise	NN	O	O
,	NN	O	O
reporter	NN	O	O
gene	NN	O	O
assays	NN	O	O
using	NN	O	O
luciferase	NN	O	O
constructs	NN	O	O
driven	NN	O	O
by	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
showed	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
controlled	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
selenium	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
selenium	NN	O	O
were	NN	O	O
specific	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
since	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
AP-1	NN	O	B-protein
was	NN	O	O
not	NN	O	O
suppressed	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
selenium	NN	O	O
supplementation	NN	O	O
may	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
modulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
p23	NN	O	B-protein
,	NN	O	O
hsp90	NN	O	B-protein
,	NN	O	O
and	NN	O	O
immunophilins	NN	O	B-protein
in	NN	O	O
the	NN	O	O
assembly	NN	O	O
of	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

To	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
assembly	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
PR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
developed	NN	O	O
cell-free	NN	O	O
systems	NN	O	O
for	NN	O	O
studying	NN	O	O
interactions	NN	O	O
of	NN	O	O
PR	NN	O	B-protein
,	NN	O	O
hsp90	NN	O	B-protein
,	NN	O	O
and	NN	O	O
other	NN	O	O
associated	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

When	NN	O	O
PR	NN	O	B-protein
is	NN	O	O
incubated	NN	O	O
in	NN	O	O
rabbit	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
,	NN	O	O
its	NN	O	O
association	NN	O	O
with	NN	O	O
hsp90	NN	O	B-protein
,	NN	O	O
hsp70	NN	O	B-protein
,	NN	O	O
the	NN	O	O
three	NN	O	O
immunophilins	NN	O	B-protein
FKBP54	NN	O	B-protein
,	NN	O	O
FKBP52	NN	O	B-protein
and	NN	O	O
CyP-40	NN	O	B-protein
,	NN	O	O
and	NN	O	O
with	NN	O	O
p23	NN	O	B-protein
is	NN	O	O
observed	NN	O	O
.	NN	O	O

These	NN	O	O
interactions	NN	O	O
require	NN	O	O
ATP/Mg2+	NN	O	O
and	NN	O	O
when	NN	O	O
ATP	NN	O	O
is	NN	O	O
limiting	NN	O	O
the	NN	O	O
PR	NN	O	B-protein
complex	NN	O	O
is	NN	O	O
altered	NN	O	O
to	NN	O	O
one	NN	O	O
containing	NN	O	O
the	NN	O	O
proteins	NN	O	O
p60	NN	O	B-protein
and	NN	O	O
p48	NN	O	B-protein
,	NN	O	O
but	NN	O	O
lacking	NN	O	O
immunophilins	NN	O	B-protein
and	NN	O	O
p23	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
two	NN	O	O
pre-formed	NN	O	B-protein
hsp90	NN	O	I-protein
complexes	NN	O	I-protein
that	NN	O	O
may	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
assembly	NN	O	O
of	NN	O	O
PR	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

One	NN	O	O
contains	NN	O	O
hsp90	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
hsp70	NN	O	B-protein
and	NN	O	O
p60	NN	O	B-protein
and	NN	O	O
this	NN	O	O
complex	NN	O	O
forms	NN	O	O
spontaneously	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
ATP	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
complex	NN	O	O
contains	NN	O	O
hsp90	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
p23	NN	O	B-protein
plus	NN	O	O
the	NN	O	O
three	NN	O	O
immunophilins	NN	O	B-protein
and	NN	O	O
some	NN	O	O
hsp70	NN	O	B-protein
.	NN	O	O

The	NN	O	O
formation	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
requires	NN	O	O
ATP	NN	O	O
.	NN	O	O

In	NN	O	O
further	NN	O	O
studies	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
purified	NN	O	O
hsp90	NN	O	B-protein
can	NN	O	O
bind	NN	O	O
to	NN	O	O
purified	NN	O	O
p23	NN	O	B-protein
and	NN	O	O
this	NN	O	O
interaction	NN	O	O
requires	NN	O	O
both	NN	O	O
ATP	NN	O	O
and	NN	O	O
molybdate	NN	O	O
.	NN	O	O

This	NN	O	O
explains	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
the	NN	O	O
known	NN	O	O
effects	NN	O	O
of	NN	O	O
ATP	NN	O	O
and	NN	O	O
molybdate	NN	O	O
on	NN	O	O
assembly	NN	O	O
of	NN	O	O
PR	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Increased	NN	O	O
IkappaB	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
diminished	NN	O	O
nuclear	NN	O	B-protein
NF-kappaB	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
following	NN	O	O
hydrocortisone	NN	O	O
injection	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
hydrocortisone	NN	O	O
and	NN	O	O
other	NN	O	O
glucocorticoids	NN	O	O
inhibit	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
generation	NN	O	O
by	NN	O	O
mononuclear	NN	O	B-cell_type
(	NN	O	I-cell_type
MNC	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	I-cell_type
polymorphonuclear	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
(	NN	O	O
PMNL	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

Since	NN	O	O
NF-kappaB/	NN	O	O
IkappaB	NN	O	B-protein
system	NN	O	O
regulates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
genes	NN	O	I-protein
,	NN	O	O
including	NN	O	O
those	NN	O	O
responsible	NN	O	O
for	NN	O	O
ROS	NN	O	O
generation	NN	O	O
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
hydrocortisone	NN	O	O
may	NN	O	O
stimulate	NN	O	O
IkappaB	NN	O	B-protein
production	NN	O	O
thus	NN	O	O
inhibiting	NN	O	O
NF-kappaB	NN	O	B-protein
translocation	NN	O	O
from	NN	O	O
the	NN	O	O
cytosol	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
in	NN	O	O
MNC	NN	O	B-cell_type
,	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

One	NN	O	O
hundred	NN	O	O
milligram	NN	O	O
of	NN	O	O
hydrocortisone	NN	O	O
was	NN	O	O
injected	NN	O	O
intravenously	NN	O	O
into	NN	O	O
4	NN	O	O
normal	NN	O	O
subjects	NN	O	O
.	NN	O	O

Blood	NN	O	O
samples	NN	O	O
were	NN	O	O
obtained	NN	O	O
prior	NN	O	O
to	NN	O	O
the	NN	O	O
injection	NN	O	O
and	NN	O	O
at	NN	O	O
1	NN	O	O
,	NN	O	O
2	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
8	NN	O	O
and	NN	O	O
24	NN	O	O
hr	NN	O	O
after	NN	O	O
the	NN	O	O
injection	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
extracts	NN	O	O
and	NN	O	O
total	NN	O	O
cell	NN	O	O
lysates	NN	O	O
were	NN	O	O
prepared	NN	O	O
from	NN	O	O
MNC	NN	O	B-cell_type
by	NN	O	O
standard	NN	O	O
techniques	NN	O	O
.	NN	O	O

IkappaB	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
MNC	NN	O	B-cell_type
homogenates	NN	O	O
increased	NN	O	O
at	NN	O	O
1	NN	O	O
hr	NN	O	O
,	NN	O	O
peaked	NN	O	O
at	NN	O	O
2-4	NN	O	O
hr	NN	O	O
,	NN	O	O
started	NN	O	O
to	NN	O	O
decrease	NN	O	O
at	NN	O	O
8	NN	O	O
hr	NN	O	O
,	NN	O	O
and	NN	O	O
returned	NN	O	O
to	NN	O	O
baseline	NN	O	O
levels	NN	O	O
at	NN	O	O
24	NN	O	O
hr	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
in	NN	O	O
MNC	NN	O	B-cell_type
nuclear	NN	O	O
extracts	NN	O	O
decreased	NN	O	O
at	NN	O	O
1	NN	O	O
hr	NN	O	O
,	NN	O	O
reached	NN	O	O
a	NN	O	O
nadir	NN	O	O
at	NN	O	O
4	NN	O	O
hr	NN	O	O
,	NN	O	O
gradually	NN	O	O
increased	NN	O	O
at	NN	O	O
8	NN	O	O
hr	NN	O	O
and	NN	O	O
returned	NN	O	O
back	NN	O	O
to	NN	O	O
baseline	NN	O	O
levels	NN	O	O
at	NN	O	O
24	NN	O	O
hr	NN	O	O
.	NN	O	O

The	NN	O	O
total	NN	O	O
protein	NN	O	O
content	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
subunit	NN	O	I-protein
(	NN	O	O
P65	NN	O	B-protein
)	NN	O	O
in	NN	O	O
MNC	NN	O	B-cell_type
lysates	NN	O	O
also	NN	O	O
showed	NN	O	O
a	NN	O	O
decrease	NN	O	O
following	NN	O	O
hydrocortisone	NN	O	O
injection	NN	O	O
.	NN	O	O

This	NN	O	O
decrease	NN	O	O
was	NN	O	O
observed	NN	O	O
at	NN	O	O
2	NN	O	O
hr	NN	O	O
,	NN	O	O
reached	NN	O	O
a	NN	O	O
nadir	NN	O	O
at	NN	O	O
4	NN	O	O
hr	NN	O	O
,	NN	O	O
and	NN	O	O
returned	NN	O	O
to	NN	O	O
baseline	NN	O	O
levels	NN	O	O
at	NN	O	O
24	NN	O	O
hr	NN	O	O
.	NN	O	O

ROS	NN	O	O
generation	NN	O	O
inhibition	NN	O	O
paralleled	NN	O	O
NF-kappaB	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
inhibited	NN	O	O
at	NN	O	O
1	NN	O	O
hr	NN	O	O
,	NN	O	O
reached	NN	O	O
a	NN	O	O
nadir	NN	O	O
at	NN	O	O
2-4	NN	O	O
hr	NN	O	O
,	NN	O	O
started	NN	O	O
to	NN	O	O
increase	NN	O	O
at	NN	O	O
8	NN	O	O
hr	NN	O	O
,	NN	O	O
and	NN	O	O
returned	NN	O	O
to	NN	O	O
basal	NN	O	O
levels	NN	O	O
at	NN	O	O
24	NN	O	O
hr	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
hydrocortisone	NN	O	O
induces	NN	O	O
IkappaB	NN	O	B-protein
and	NN	O	O
suppresses	NN	O	O
NF-kappaB	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
MNC	NN	O	B-cell_type
in	NN	O	O
parallel	NN	O	O
.	NN	O	O

IkappaB	NN	O	B-protein
further	NN	O	O
reduces	NN	O	O
the	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
thus	NN	O	O
preventing	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Glucocorticoids	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
:	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
.	NN	O	O

Glucocorticoids	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
are	NN	O	O
potent	NN	O	O
anti-inflammatory	NN	O	O
and	NN	O	O
immunosuppressive	NN	O	O
agents	NN	O	O
that	NN	O	O
act	NN	O	O
on	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
including	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
exact	NN	O	O
cellular	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
this	NN	O	O
anti-inflammatory	NN	O	O
capacity	NN	O	O
are	NN	O	O
still	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
our	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
GC	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
were	NN	O	O
isolated	NN	O	O
by	NN	O	O
density	NN	O	O
centrifugation	NN	O	O
methods	NN	O	O
with	NN	O	O
a	NN	O	O
purity	NN	O	O
of	NN	O	O
>	NN	O	O
90	NN	O	O
%	NN	O	O
and	NN	O	O
were	NN	O	O
cultured	NN	O	O
in	NN	O	O
RPMI	NN	O	O
1640	NN	O	O
medium	NN	O	O
.	NN	O	O

Monocyte	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
four	NN	O	O
independent	NN	O	O
methods	NN	O	O
,	NN	O	O
including	NN	O	O
annexin-V	NN	O	O
staining	NN	O	O
,	NN	O	O
TUNEL	NN	O	O
,	NN	O	O
DNA-laddering	NN	O	O
,	NN	O	O
and	NN	O	O
typical	NN	O	O
morphology	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
transmission	NN	O	O
electron	NN	O	O
microscopy	NN	O	O
.	NN	O	O

TNF-alpha	NN	O	B-protein
and	NN	O	O
IL-1beta	NN	O	B-protein
were	NN	O	O
measured	NN	O	O
by	NN	O	O
ELISA	NN	O	O
.	NN	O	O

GC	NN	O	B-protein
receptor	NN	O	I-protein
was	NN	O	O
blocked	NN	O	O
with	NN	O	O
mifepristone	NN	O	O
.	NN	O	O

Caspase	NN	O	O
3	NN	O	O
was	NN	O	O
inhibited	NN	O	O
with	NN	O	O
caspase-3	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
DEVD-CHO	NN	O	O
)	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
with	NN	O	O
different	NN	O	O
GC	NN	O	O
at	NN	O	O
therapeutic	NN	O	O
concentrations	NN	O	O
resulted	NN	O	O
in	NN	O	O
monocyte	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
a	NN	O	O
time-	NN	O	O
and	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Necrosis	NN	O	O
was	NN	O	O
excluded	NN	O	O
by	NN	O	O
propidium	NN	O	O
iodide	NN	O	O
staining	NN	O	O
.	NN	O	O

Proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
such	NN	O	O
as	NN	O	O
IL-1beta	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
were	NN	O	O
down-regulated	NN	O	O
by	NN	O	O
GC	NN	O	O
treatment	NN	O	O
.	NN	O	O

Continuous	NN	O	O
treatment	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
with	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
could	NN	O	O
almost	NN	O	O
completely	NN	O	O
prevent	NN	O	O
GC-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
mifepristone	NN	O	O
or	NN	O	O
caspase-3	NN	O	O
inhibitor	NN	O	O
could	NN	O	O
partially	NN	O	O
abrogate	NN	O	O
GC-induced	NN	O	O
apoptosis	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
GC-induced	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
study	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
GC	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

GC-induced	NN	O	O
monocyte	NN	O	O
apoptosis	NN	O	O
may	NN	O	O
be	NN	O	O
partially	NN	O	O
mediated	NN	O	O
through	NN	O	O
effects	NN	O	O
on	NN	O	O
IL-1beta	NN	O	B-protein
production	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
conceivable	NN	O	O
that	NN	O	O
GC	NN	O	O
exert	NN	O	O
their	NN	O	O
anti-inflammatory	NN	O	O
capacity	NN	O	O
in	NN	O	O
various	NN	O	O
diseases	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
effects	NN	O	O
of	NN	O	O
lipopolysaccharide	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
on	NN	O	O
monocytic	NN	O	B-protein
IkappaB	NN	O	I-protein
kinase	NN	O	I-protein
signalsome	NN	O	O
activation	NN	O	O
and	NN	O	O
IkappaB	NN	O	B-protein
proteolysis	NN	O	O
.	NN	O	O

The	NN	O	O
inflammatory	NN	O	O
mediators	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
are	NN	O	O
potent	NN	O	O
activators	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

This	NN	O	O
study	NN	O	O
compared	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
these	NN	O	O
stimuli	NN	O	O
on	NN	O	O
endogenous	NN	O	O
IkappaB	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
IKK	NN	O	B-protein
)	NN	O	O
signalsome	NN	O	O
activation	NN	O	O
and	NN	O	O
IkappaB	NN	O	B-protein
phosphorylation/proteolysis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
signalsome	NN	O	B-protein
proteins	NN	O	I-protein
IKK-alpha	NN	O	B-protein
,	NN	O	O
IKK-beta	NN	O	B-protein
,	NN	O	O
NF-kappaB-inducing	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
NIK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
IKK-gamma	NN	O	B-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
essential	NN	O	O
modulator	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
IKK	NN	O	B-protein
complex-associated	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Kinase	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
TNF	NN	O	B-protein
elicited	NN	O	O
a	NN	O	O
rapid	NN	O	O
but	NN	O	O
short-lived	NN	O	O
induction	NN	O	O
of	NN	O	O
IKK	NN	O	B-protein
activity	NN	O	O
with	NN	O	O
a	NN	O	O
3-fold	NN	O	O
greater	NN	O	O
effect	NN	O	O
on	NN	O	O
IKK-alpha	NN	O	B-protein
than	NN	O	O
on	NN	O	O
IKK-beta	NN	O	B-protein
,	NN	O	O
peaking	NN	O	O
at	NN	O	O
5	NN	O	O
min	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
LPS	NN	O	O
predominantly	NN	O	O
stimulated	NN	O	O
IKK-beta	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
which	NN	O	O
slowly	NN	O	O
increased	NN	O	O
,	NN	O	O
peaking	NN	O	O
at	NN	O	O
30	NN	O	O
min	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
peak	NN	O	O
was	NN	O	O
observed	NN	O	O
at	NN	O	O
a	NN	O	O
later	NN	O	O
time	NN	O	O
point	NN	O	O
following	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
,	NN	O	O
which	NN	O	O
consisted	NN	O	O
of	NN	O	O
both	NN	O	O
IKK-alpha	NN	O	O
and	NN	O	O
-beta	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
endogenous	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
signalsome	NN	O	O
components	NN	O	O
were	NN	O	O
unaffected	NN	O	O
by	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
our	NN	O	O
studies	NN	O	O
showed	NN	O	O
association	NN	O	O
of	NN	O	O
the	NN	O	O
IKK-alpha/beta	NN	O	B-protein
heterodimer	NN	O	I-protein
with	NN	O	O
NIK	NN	O	B-protein
,	NN	O	O
IkappaB-alpha	NN	O	B-protein
and	NN	O	O
-epsilon	NN	O	B-protein
in	NN	O	O
unstimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Exposure	NN	O	O
to	NN	O	O
LPS	NN	O	O
or	NN	O	O
TNF	NN	O	B-protein
led	NN	O	O
to	NN	O	O
differential	NN	O	O
patterns	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
and	NN	O	O
IkappaB-epsilon	NN	O	B-protein
disappearance	NN	O	O
from	NN	O	O
and	NN	O	O
reassembly	NN	O	O
with	NN	O	O
the	NN	O	O
signalsome	NN	O	O
,	NN	O	O
whereas	NN	O	O
IKK-alpha	NN	O	B-protein
,	NN	O	O
IKK-beta	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NIK	NN	O	B-protein
remained	NN	O	O
complex-associated	NN	O	O
.	NN	O	O

NIK	NN	O	B-protein
can	NN	O	O
not	NN	O	O
phosphorylate	NN	O	O
IkappaB-alpha	NN	O	B-protein
directly	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
functionally	NN	O	O
important	NN	O	O
subunit	NN	O	O
,	NN	O	O
because	NN	O	O
mutated	NN	O	O
NIK	NN	O	B-protein
inhibited	NN	O	O
stimulus-induced	NN	O	O
kappaB	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
more	NN	O	O
effectively	NN	O	O
than	NN	O	O
mutated	NN	O	O
IKK-alpha	NN	O	B-protein
or	NN	O	O
-beta	NN	O	B-protein
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
IKK	NN	O	B-protein
complex-associated	NN	O	I-protein
protein	NN	O	I-protein
inhibited	NN	O	O
stimulus-mediated	NN	O	O
transcription	NN	O	O
,	NN	O	O
whereas	NN	O	O
NF-kappaB	NN	O	B-protein
essential	NN	O	O
modulator	NN	O	O
enhanced	NN	O	O
it	NN	O	O
.	NN	O	O

The	NN	O	O
understanding	NN	O	O
of	NN	O	O
LPS-	NN	O	O
and	NN	O	O
TNF-induced	NN	O	O
signaling	NN	O	O
may	NN	O	O
allow	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
specific	NN	O	O
strategies	NN	O	O
to	NN	O	O
treat	NN	O	O
sepsis-associated	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
decreases	NN	O	O
,	NN	O	O
and	NN	O	O
interleukin-10	NN	O	B-protein
increases	NN	O	O
,	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
to	NN	O	O
dexamethasone	NN	O	O
:	NN	O	O
potential	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Resistance	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
autoimmune/inflammatory	NN	O	O
diseases	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
process	NN	O	O
itself	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
examine	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNFalpha	NN	O	B-protein
,	NN	O	O
a	NN	O	O
proinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
)	NN	O	O
and	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-10	NN	O	I-protein
(	NN	O	O
an	NN	O	O
anti-inflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
)	NN	O	O
to	NN	O	O
differentially	NN	O	O
regulate	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
/macrophages	NN	O	B-cell_type
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

To	NN	O	O
accomplish	NN	O	O
this	NN	O	O
,	NN	O	O
we	NN	O	O
first	NN	O	O
analyzed	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
TNFalpha	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	O
inhibition	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
in	NN	O	O
LPS-stimulated	NN	O	B-cell_line
whole-blood	NN	O	I-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

Second	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
by	NN	O	O
preincubation	NN	O	O
with	NN	O	O
TNFalpha	NN	O	B-protein
or	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
measurement	NN	O	O
of	NN	O	O
LPS-stimulated	NN	O	O
IL-6	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
evaluated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
on	NN	O	O
phorbolmyristate-acetate-stimulated	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	I-protein
antagonist	NN	O	O
secretion	NN	O	O
by	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U937	NN	O	I-cell_line
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
corticosensitivity	NN	O	O
in	NN	O	O
TNFalpha-	NN	O	B-cell_line
and	NN	O	I-cell_line
IL-10-pretreated	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
related	NN	O	O
to	NN	O	O
a	NN	O	O
change	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentration	NN	O	O
and	NN	O	O
affinity	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
had	NN	O	O
different	NN	O	O
effects	NN	O	O
on	NN	O	O
LPS-induced	NN	O	O
TNFalpha	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
secretion	NN	O	O
;	NN	O	O
whereas	NN	O	O
it	NN	O	O
suppressed	NN	O	O
TNFalpha	NN	O	B-protein
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
fashion	NN	O	O
,	NN	O	O
its	NN	O	O
effect	NN	O	O
on	NN	O	O
IL-10	NN	O	B-protein
secretion	NN	O	O
was	NN	O	O
biphasic	NN	O	O
,	NN	O	O
producing	NN	O	O
stimulation	NN	O	O
at	NN	O	O
lower	NN	O	O
,	NN	O	O
and	NN	O	O
inhibition	NN	O	O
at	NN	O	O
higher	NN	O	O
doses	NN	O	O
.	NN	O	O

The	NN	O	O
concentration	NN	O	O
of	NN	O	O
LPS	NN	O	O
employed	NN	O	O
influenced	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
on	NN	O	O
IL-10	NN	O	B-protein
secretion	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
with	NN	O	O
TNFalpha	NN	O	B-protein
diminished	NN	O	O
,	NN	O	O
and	NN	O	O
with	NN	O	O
IL-10	NN	O	B-protein
improved	NN	O	O
,	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
to	NN	O	O
suppress	NN	O	O
IL-6	NN	O	B-protein
secretion	NN	O	O
in	NN	O	O
whole-blood	NN	O	B-cell_type
cell	NN	O	I-cell_type
cultures	NN	O	I-cell_type
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
for	NN	O	O
both	NN	O	O
)	NN	O	O
and	NN	O	O
to	NN	O	O
enhance	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	O
antagonist	NN	O	O
secretion	NN	O	O
by	NN	O	O
U937	NN	O	O
cells	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
for	NN	O	O
both	NN	O	O
)	NN	O	O
.	NN	O	O

TNFalpha	NN	O	B-protein
decreased	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
,	NN	O	O
while	NN	O	O
IL-10	NN	O	B-protein
increased	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
sites	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
with	NN	O	O
no	NN	O	O
discernible	NN	O	O
effect	NN	O	O
on	NN	O	O
their	NN	O	O
binding	NN	O	O
affinity	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
differentially	NN	O	O
modulate	NN	O	O
TNFalpha	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
in	NN	O	O
a	NN	O	O
LPS	NN	O	O
dose-dependent	NN	O	O
fashion	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
is	NN	O	O
altered	NN	O	O
by	NN	O	O
TNFalpha	NN	O	B-protein
or	NN	O	O
IL-10	NN	O	B-protein
pretreatment	NN	O	O
;	NN	O	O
TNFalpha	NN	O	B-protein
blocks	NN	O	O
their	NN	O	O
effects	NN	O	O
,	NN	O	O
whereas	NN	O	O
IL-10	NN	O	B-protein
acts	NN	O	O
synergistically	NN	O	O
with	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
opposite	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
changes	NN	O	O
,	NN	O	O
respectively	NN	O	O
opposing	NN	O	O
and	NN	O	O
favoring	NN	O	O
glucocorticoid	NN	O	O
actions	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
pro-/antiinflammatory	NN	O	O
cytokine	NN	O	B-protein
secretion	NN	O	O
may	NN	O	O
alter	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
patients	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Block	NN	O	O
of	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
32Dcl3	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
AML1/ETO	NN	O	B-DNA
(	NN	O	I-DNA
MTG8	NN	O	I-DNA
)	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
by	NN	O	O
highly	NN	O	O
expressed	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
chimeric	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	I-DNA
AML1/ETO	NN	O	B-DNA
(	NN	O	I-DNA
MTG8	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
generated	NN	O	O
in	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
8	NN	O	I-DNA
;	NN	O	I-DNA
21	NN	O	I-DNA
)	NN	O	I-DNA
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
enhances	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
whether	NN	O	O
this	NN	O	O
enhancement	NN	O	O
is	NN	O	O
the	NN	O	O
primary	NN	O	O
role	NN	O	O
of	NN	O	O
AML1/ETO	NN	O	B-DNA
in	NN	O	O
leukemogenesis	NN	O	O
,	NN	O	O
effects	NN	O	O
of	NN	O	O
over-expression	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
precursor	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
32Dcl3	NN	O	B-cell_line
,	NN	O	O
were	NN	O	O
examined	NN	O	O
.	NN	O	O

When	NN	O	O
32Dcl3	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
exogenous	NN	O	O
Bcl-2	NN	O	B-protein
were	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
,	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
morphological	NN	O	O
differentiation	NN	O	O
was	NN	O	O
delayed	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
even	NN	O	O
the	NN	O	O
cells	NN	O	O
expressing	NN	O	O
very	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
exogenous	NN	O	O
Bcl-2	NN	O	B-protein
eventually	NN	O	O
underwent	NN	O	O
differentiation	NN	O	O
without	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

On	NN	O	O
the	NN	O	O
contrary	NN	O	O
,	NN	O	O
32Dcl3	NN	O	B-cell_line
cells	NN	O	I-cell_line
stably	NN	O	O
expressing	NN	O	O
AML1/ETO	NN	O	B-DNA
were	NN	O	O
completely	NN	O	O
resistant	NN	O	O
to	NN	O	O
differentiation	NN	O	O
and	NN	O	O
continued	NN	O	O
to	NN	O	O
grow	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
G-CSF	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
interpretation	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
not	NN	O	O
the	NN	O	O
primary	NN	O	O
target	NN	O	O
of	NN	O	O
AML1/ETO	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Host	NN	O	O
defense	NN	O	O
mechanisms	NN	O	O
triggered	NN	O	O
by	NN	O	O
microbial	NN	O	B-protein
lipoproteins	NN	O	I-protein
through	NN	O	O
toll-like	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
generation	NN	O	O
of	NN	O	O
cell-mediated	NN	O	O
immunity	NN	O	O
against	NN	O	O
many	NN	O	O
infectious	NN	O	O
pathogens	NN	O	O
involves	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
interleukin-12	NN	O	B-protein
(	NN	O	O
IL-12	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
key	NN	O	O
signal	NN	O	O
of	NN	O	O
the	NN	O	O
innate	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Yet	NN	O	O
,	NN	O	O
for	NN	O	O
many	NN	O	O
pathogens	NN	O	O
,	NN	O	O
the	NN	O	O
molecules	NN	O	O
that	NN	O	O
induce	NN	O	O
IL-12	NN	O	B-protein
production	NN	O	O
by	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
they	NN	O	O
do	NN	O	O
so	NN	O	O
remain	NN	O	O
undefined	NN	O	O
.	NN	O	O

Here	NN	O	O
it	NN	O	O
is	NN	O	O
shown	NN	O	O
that	NN	O	O
microbial	NN	O	B-protein
lipoproteins	NN	O	I-protein
are	NN	O	O
potent	NN	O	O
stimulators	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
production	NN	O	O
by	NN	O	O
human	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
that	NN	O	O
induction	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
Toll-like	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
TLRs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Several	NN	O	O
lipoproteins	NN	O	O
stimulated	NN	O	O
TLR-dependent	NN	O	O
transcription	NN	O	O
of	NN	O	O
inducible	NN	O	O
nitric	NN	O	B-protein
oxide	NN	O	I-protein
synthase	NN	O	I-protein
and	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
,	NN	O	O
a	NN	O	O
powerful	NN	O	O
microbicidal	NN	O	O
pathway	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
TLRs	NN	O	B-protein
by	NN	O	O
microbial	NN	O	B-protein
lipoproteins	NN	O	I-protein
may	NN	O	O
initiate	NN	O	O
innate	NN	O	O
defense	NN	O	O
mechanisms	NN	O	O
against	NN	O	O
infectious	NN	O	O
pathogens	NN	O	O
.	NN	O	O

-DOCSTART-	O

Abnormal	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
decreased	NN	O	O
p65-RelA	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
.	NN	O	O

Numerous	NN	O	O
cellular	NN	O	O
and	NN	O	O
biochemical	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
immune	NN	O	O
regulation	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
(	NN	O	O
SLE	NN	O	O
)	NN	O	O
,	NN	O	O
including	NN	O	O
surface	NN	O	O
Ag	NN	O	B-protein
receptor	NN	O	I-protein
-initiated	NN	O	O
signaling	NN	O	O
events	NN	O	O
and	NN	O	O
lymphokine	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Because	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
numerous	NN	O	O
inflammatory	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
molecular	NN	O	O
target	NN	O	O
of	NN	O	O
antiinflammatory	NN	O	O
drugs	NN	O	O
,	NN	O	O
we	NN	O	O
sought	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
functional	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
protein	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
lupus	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Freshly	NN	O	O
isolated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
lupus	NN	O	O
patients	NN	O	O
,	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
patients	NN	O	O
,	NN	O	O
and	NN	O	O
normal	NN	O	O
individuals	NN	O	O
were	NN	O	O
activated	NN	O	O
physiologically	NN	O	O
via	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
with	NN	O	O
anti-CD3	NN	O	B-protein
and	NN	O	O
anti-CD28	NN	O	B-protein
Abs	NN	O	I-protein
to	NN	O	O
assess	NN	O	O
proximal	NN	O	O
membrane	NN	O	O
signaling	NN	O	O
,	NN	O	O
and	NN	O	O
with	NN	O	O
PMA	NN	O	O
and	NN	O	O
a	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
(	NN	O	O
A23187	NN	O	O
)	NN	O	O
to	NN	O	O
bypass	NN	O	O
membrane-mediated	NN	O	O
signaling	NN	O	O
events	NN	O	O
.	NN	O	O

We	NN	O	O
measured	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
by	NN	O	O
gel	NN	O	O
shift	NN	O	O
analysis	NN	O	O
.	NN	O	O

When	NN	O	O
compared	NN	O	O
with	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
SLE	NN	O	O
patients	NN	O	O
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
in	NN	O	O
SLE	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
RA	NN	O	O
,	NN	O	O
patients	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
absent	NN	O	O
in	NN	O	O
several	NN	O	O
SLE	NN	O	O
patients	NN	O	O
who	NN	O	O
were	NN	O	O
not	NN	O	O
receiving	NN	O	O
any	NN	O	O
medication	NN	O	O
,	NN	O	O
including	NN	O	O
corticosteroids	NN	O	O
.	NN	O	O

Also	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
remained	NN	O	O
absent	NN	O	O
in	NN	O	O
follow-up	NN	O	O
studies	NN	O	O
.	NN	O	O

In	NN	O	O
supershift	NN	O	O
experiments	NN	O	O
using	NN	O	O
specific	NN	O	O
Abs	NN	O	B-protein
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
group	NN	O	O
of	NN	O	O
SLE	NN	O	O
patients	NN	O	O
who	NN	O	O
displayed	NN	O	O
undetectable	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
p65	NN	O	B-protein
complexes	NN	O	I-protein
were	NN	O	O
not	NN	O	O
formed	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
immunoblot	NN	O	O
analysis	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
showed	NN	O	O
decreased	NN	O	O
or	NN	O	O
absent	NN	O	O
p65	NN	O	B-protein
protein	NN	O	I-protein
levels	NN	O	O
.	NN	O	O

As	NN	O	O
p65	NN	O	B-protein
complexes	NN	O	I-protein
are	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
in	NN	O	O
comparison	NN	O	O
to	NN	O	O
the	NN	O	O
p50	NN	O	O
homodimer	NN	O	O
,	NN	O	O
this	NN	O	O
novel	NN	O	O
finding	NN	O	O
may	NN	O	O
provide	NN	O	O
insight	NN	O	O
on	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
abnormal	NN	O	B-protein
cytokine	NN	O	I-protein
or	NN	O	O
other	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
SLE	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signaling	NN	O	O
events	NN	O	O
induced	NN	O	O
by	NN	O	O
lipopolysaccharide-activated	NN	O	B-protein
toll-like	NN	O	I-protein
receptor	NN	O	I-protein
2	NN	O	I-protein
.	NN	O	O

Human	NN	O	B-protein
Toll-like	NN	O	I-protein
receptor	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	I-protein
TLR2	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
a	NN	O	O
signaling	NN	O	B-protein
receptor	NN	O	I-protein
that	NN	O	O
responds	NN	O	O
to	NN	O	O
LPS	NN	O	O
and	NN	O	O
activates	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
investigate	NN	O	O
further	NN	O	O
the	NN	O	O
events	NN	O	O
triggered	NN	O	O
by	NN	O	O
TLR2	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
TLR2	NN	O	B-protein
associates	NN	O	O
with	NN	O	O
the	NN	O	O
high-affinity	NN	O	B-protein
LPS	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
membrane	NN	O	O
CD14	NN	O	B-protein
to	NN	O	O
serve	NN	O	O
as	NN	O	O
an	NN	O	O
LPS	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
LPS	NN	O	O
treatment	NN	O	O
enhances	NN	O	O
the	NN	O	O
oligomerization	NN	O	O
of	NN	O	O
TLR2	NN	O	B-protein
.	NN	O	O

Concomitant	NN	O	O
with	NN	O	O
receptor	NN	O	O
oligomerization	NN	O	O
,	NN	O	O
the	NN	O	O
IL-1R-associated	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
IRAK	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
recruited	NN	O	O
to	NN	O	O
the	NN	O	O
TLR2	NN	O	B-protein
complex	NN	O	O
.	NN	O	O

Intracellular	NN	O	O
deletion	NN	O	O
variants	NN	O	O
of	NN	O	O
TLR2	NN	O	B-protein
lacking	NN	O	O
C-terminal	NN	O	O
13	NN	O	O
or	NN	O	O
141	NN	O	O
aa	NN	O	O
fail	NN	O	O
to	NN	O	O
recruit	NN	O	O
IRAK	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
inability	NN	O	O
of	NN	O	O
these	NN	O	O
mutants	NN	O	O
to	NN	O	O
transmit	NN	O	O
LPS	NN	O	O
cellular	NN	O	O
signaling	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
both	NN	O	O
deletion	NN	O	O
mutants	NN	O	O
could	NN	O	O
still	NN	O	O
form	NN	O	O
complexes	NN	O	O
with	NN	O	O
wild-type	NN	O	B-protein
TLR2	NN	O	I-protein
and	NN	O	O
act	NN	O	O
in	NN	O	O
a	NN	O	O
dominant-negative	NN	O	O
(	NN	O	O
DN	NN	O	O
)	NN	O	O
fashion	NN	O	O
to	NN	O	O
block	NN	O	O
TLR2	NN	O	B-protein
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

DN	NN	O	O
constructs	NN	O	O
of	NN	O	O
myeloid	NN	O	B-protein
differentiation	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
IRAK	NN	O	B-protein
,	NN	O	O
TNF	NN	O	B-protein
receptor-associated	NN	O	I-protein
factor	NN	O	I-protein
6	NN	O	I-protein
,	NN	O	O
and	NN	O	O
NF-kappaB-inducing	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
when	NN	O	O
coexpressed	NN	O	O
with	NN	O	O
TLR2	NN	O	B-protein
,	NN	O	O
abrogate	NN	O	O
TLR2	NN	O	B-protein
-mediated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
reveal	NN	O	O
a	NN	O	O
conserved	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
for	NN	O	O
TLR2	NN	O	B-protein
and	NN	O	O
IL-1Rs	NN	O	B-protein
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
TLR2	NN	O	B-protein
by	NN	O	O
DN	NN	O	O
variants	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tyrphostin	NN	O	O
AG-490	NN	O	O
inhibits	NN	O	O
cytokine-mediated	NN	O	O
JAK3	NN	O	B-protein
/STAT5a/b	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
and	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
of	NN	O	O
antigen-activated	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Janus	NN	O	B-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	I-protein
JAK3	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
required	NN	O	O
for	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
development	NN	O	O
and	NN	O	O
activated	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
utilize	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	I-protein
IL-2	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
common	NN	O	I-protein
gamma	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	I-protein
gamma	NN	O	I-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Genetic	NN	O	O
inactivation	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
is	NN	O	O
manifested	NN	O	O
as	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
disease	NN	O	O
(	NN	O	O
SCID	NN	O	O
)	NN	O	O
in	NN	O	O
humans	NN	O	O
and	NN	O	O
mice	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
JAK3	NN	O	B-protein
represents	NN	O	O
a	NN	O	O
pharmacological	NN	O	O
target	NN	O	O
to	NN	O	O
control	NN	O	O
certain	NN	O	O
lymphoid-derived	NN	O	O
diseases	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
provide	NN	O	O
novel	NN	O	O
evidence	NN	O	O
that	NN	O	O
AG-490	NN	O	O
potently	NN	O	O
inhibits	NN	O	O
the	NN	O	O
autokinase	NN	O	O
activity	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
and	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
5a	NN	O	I-protein
and	NN	O	I-protein
5b	NN	O	I-protein
(	NN	O	I-protein
STAT5a/b	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Similar	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
were	NN	O	O
observed	NN	O	O
with	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
use	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

AG-490	NN	O	O
also	NN	O	O
inhibited	NN	O	O
IL-2	NN	O	B-protein
-mediated	NN	O	O
proliferative	NN	O	O
growth	NN	O	O
in	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
an	NN	O	O
IC50	NN	O	O
)	NN	O	O
=	NN	O	O
25	NN	O	O
microM	NN	O	O
that	NN	O	O
was	NN	O	O
partially	NN	O	O
recoverable	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
this	NN	O	O
inhibitor	NN	O	O
prevented	NN	O	O
tetanus	NN	O	O
toxoid	NN	O	O
antigen-specific	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
expansion	NN	O	O
but	NN	O	O
failed	NN	O	O
to	NN	O	O
block	NN	O	O
activation	NN	O	O
of	NN	O	O
Zap70	NN	O	B-protein
or	NN	O	O
p56Lck	NN	O	B-protein
after	NN	O	O
anti-CD3	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
AG-490	NN	O	O
inhibits	NN	O	O
the	NN	O	O
JAK3	NN	O	B-protein
-mediated	NN	O	O
Type	NN	O	O
II	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
receptor-derived	NN	O	O
Type	NN	O	O
I	NN	O	O
pathway	NN	O	O
and	NN	O	O
possesses	NN	O	O
therapeutic	NN	O	O
potential	NN	O	O
for	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
-derived	NN	O	O
pathologies	NN	O	O
such	NN	O	O
as	NN	O	O
graft-versus-host	NN	O	O
disease	NN	O	O
,	NN	O	O
allergy	NN	O	O
,	NN	O	O
and	NN	O	O
autoimmune	NN	O	O
disorders	NN	O	O
.	NN	O	O

-DOCSTART-	O

C/EBP	NN	O	B-protein
beta	NN	O	I-protein
in	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
:	NN	O	O
correlation	NN	O	O
with	NN	O	O
inflammation	NN	O	O
,	NN	O	O
not	NN	O	O
disease	NN	O	O
specificity	NN	O	O
.	NN	O	O

Rheumatoid	NN	O	O
arthritis	NN	O	O
synovial	NN	O	O
tissue	NN	O	O
was	NN	O	O
examined	NN	O	O
and	NN	O	O
compared	NN	O	O
with	NN	O	O
osteoarthritis	NN	O	O
tissue	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
C/EBP	NN	O	I-protein
beta	NN	O	I-protein
(	NN	O	I-protein
NF-IL-6	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

The	NN	O	O
region	NN	O	O
(	NN	O	O
lining	NN	O	O
or	NN	O	O
sublining	NN	O	O
)	NN	O	O
,	NN	O	O
cell	NN	O	O
type	NN	O	O
,	NN	O	O
and	NN	O	O
subcellular	NN	O	O
distribution	NN	O	O
(	NN	O	O
cytoplasmic	NN	O	O
or	NN	O	O
nuclear	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
was	NN	O	O
characterized	NN	O	O
.	NN	O	O

Rheumatoid	NN	O	O
arthritis	NN	O	O
synovial	NN	O	O
fluid	NN	O	O
and	NN	O	O
blood	NN	O	O
and	NN	O	O
normal	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
were	NN	O	O
also	NN	O	O
examined	NN	O	O
.	NN	O	O

C/EBP	NN	O	B-protein
beta	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
synovial	NN	O	O
lining	NN	O	O
and	NN	O	O
in	NN	O	O
sublining	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	I-cell_type
synovial	NN	O	I-cell_type
tissue	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
both	NN	O	O
rheumatoid	NN	O	O
and	NN	O	O
osteoarthritis	NN	O	O
.	NN	O	O

A	NN	O	O
significant	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
and	NN	O	O
<	NN	O	O
0.05	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
nuclear	NN	O	O
staining	NN	O	O
was	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
lining	NN	O	O
layer	NN	O	O
,	NN	O	O
compared	NN	O	O
to	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
sublining	NN	O	O
region	NN	O	O
,	NN	O	O
in	NN	O	O
rheumatoid	NN	O	O
and	NN	O	O
osteoarthritis	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
diseases	NN	O	O
a	NN	O	O
strong	NN	O	O
correlation	NN	O	O
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.79	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
synovial	NN	O	O
lining	NN	O	O
that	NN	O	O
were	NN	O	O
positive	NN	O	O
for	NN	O	O
nuclear	NN	O	B-protein
C/EBP	NN	O	I-protein
beta	NN	O	I-protein
and	NN	O	O
lining	NN	O	O
cell	NN	O	O
depth	NN	O	O
.	NN	O	O

Two-color	NN	O	O
immunohistochemistry	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
both	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
fibroblast-like	NN	O	B-cell_type
synoviocytes	NN	O	I-cell_type
were	NN	O	O
positive	NN	O	O
for	NN	O	O
nuclear	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
immunohistochemistry	NN	O	O
and	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
with	NN	O	O
isolated	NN	O	O
synovial	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

Nuclear	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
was	NN	O	O
also	NN	O	O
detected	NN	O	O
in	NN	O	O
rheumatoid	NN	O	B-cell_type
synovial	NN	O	I-cell_type
fluid	NN	O	I-cell_type
monocytes/macrophages	NN	O	I-cell_type
,	NN	O	I-cell_type
but	NN	O	I-cell_type
not	NN	O	I-cell_type
in	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
or	NN	O	I-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
confirmed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
intensity	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
staining	NN	O	O
was	NN	O	O
greater	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
in	NN	O	O
synovial	NN	O	B-cell_type
fluid	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
than	NN	O	O
in	NN	O	O
those	NN	O	O
from	NN	O	O
normal	NN	O	B-cell_type
or	NN	O	I-cell_type
rheumatoid	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
the	NN	O	O
enhanced	NN	O	O
nuclear	NN	O	O
staining	NN	O	O
for	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
in	NN	O	O
the	NN	O	O
synovial	NN	O	O
lining	NN	O	O
,	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
sublining	NN	O	O
,	NN	O	O
suggesting	NN	O	O
activation	NN	O	O
in	NN	O	O
the	NN	O	O
lining	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
positive	NN	O	O
correlation	NN	O	O
of	NN	O	O
lining	NN	O	O
layer	NN	O	O
depth	NN	O	O
with	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
lining	NN	O	O
positive	NN	O	O
for	NN	O	O
nuclear	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
suggest	NN	O	O
a	NN	O	O
potential	NN	O	O
role	NN	O	O
for	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
in	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
production	NN	O	O
or	NN	O	O
activity	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
to	NN	O	O
inhibit	NN	O	O
inflammatory	NN	O	O
mediator	NN	O	O
expression	NN	O	O
by	NN	O	O
synovial	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
fibroblasts	NN	O	B-cell_type
,	NN	O	O
offers	NN	O	O
a	NN	O	O
novel	NN	O	O
approach	NN	O	O
to	NN	O	O
therapeutic	NN	O	O
intervention	NN	O	O
.	NN	O	O

-DOCSTART-	O

Direct	NN	O	O
interaction	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
PU.1	NN	O	I-protein
and	NN	O	I-protein
GATA-1	NN	O	I-protein
:	NN	O	O
functional	NN	O	O
antagonism	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Malignant	NN	O	O
transformation	NN	O	O
usually	NN	O	O
inhibits	NN	O	O
terminal	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
but	NN	O	O
the	NN	O	O
precise	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
are	NN	O	O
not	NN	O	O
understood	NN	O	O
.	NN	O	O

PU.1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
hematopoietic-specific	NN	O	B-protein
Ets	NN	O	I-protein
family	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
development	NN	O	O
of	NN	O	O
some	NN	O	O
lymphoid	NN	O	B-cell_type
and	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

PU.1	NN	O	B-protein
can	NN	O	O
also	NN	O	O
act	NN	O	O
as	NN	O	O
an	NN	O	O
oncoprotein	NN	O	B-protein
as	NN	O	O
activation	NN	O	O
of	NN	O	O
its	NN	O	O
expression	NN	O	O
in	NN	O	O
erythroid	NN	O	O
precursors	NN	O	O
by	NN	O	O
proviral	NN	O	O
insertion	NN	O	O
or	NN	O	O
transgenesis	NN	O	O
causes	NN	O	O
erythroleukemias	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

Restoration	NN	O	O
of	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	B-cell_line
erythroleukemia	NN	O	I-cell_line
(	NN	O	I-cell_line
MEL	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
requires	NN	O	O
a	NN	O	O
decline	NN	O	O
in	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
PU.1	NN	O	B-protein
can	NN	O	O
block	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
investigate	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
PU.1	NN	O	B-protein
interferes	NN	O	O
with	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
PU.1	NN	O	B-protein
interacts	NN	O	O
directly	NN	O	O
with	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
required	NN	O	O
for	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Interaction	NN	O	O
between	NN	O	O
PU.1	NN	O	B-protein
and	NN	O	O
GATA-1	NN	O	B-protein
requires	NN	O	O
intact	NN	O	O
DNA-binding	NN	O	B-protein
domains	NN	O	I-protein
in	NN	O	O
both	NN	O	O
proteins	NN	O	O
.	NN	O	O

PU.1	NN	O	B-protein
represses	NN	O	O
GATA-1	NN	O	B-protein
-mediated	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Both	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
and	NN	O	O
transactivation	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
PU.1	NN	O	B-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
repression	NN	O	O
and	NN	O	O
both	NN	O	O
domains	NN	O	O
are	NN	O	O
also	NN	O	O
needed	NN	O	O
to	NN	O	O
block	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
in	NN	O	O
MEL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
PU.1	NN	O	B-protein
in	NN	O	O
Xenopus	NN	O	O
embryos	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
block	NN	O	O
erythropoiesis	NN	O	O
during	NN	O	O
normal	NN	O	O
development	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
introduction	NN	O	O
of	NN	O	O
exogenous	NN	O	O
GATA-1	NN	O	B-protein
in	NN	O	O
both	NN	O	O
MEL	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
Xenopus	NN	O	O
embryos	NN	O	O
and	NN	O	O
explants	NN	O	O
relieves	NN	O	O
the	NN	O	O
block	NN	O	O
to	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
imposed	NN	O	O
by	NN	O	O
PU.1	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
stoichiometry	NN	O	O
of	NN	O	O
directly	NN	O	O
interacting	NN	O	O
but	NN	O	O
opposing	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
may	NN	O	O
be	NN	O	O
a	NN	O	O
crucial	NN	O	O
determinant	NN	O	O
governing	NN	O	O
processes	NN	O	O
of	NN	O	O
normal	NN	O	O
differentiation	NN	O	O
and	NN	O	O
malignant	NN	O	O
transformation	NN	O	O
.	NN	O	O

-DOCSTART-	O

IL-12	NN	O	B-protein
induces	NN	O	O
IFN	NN	O	B-protein
regulating	NN	O	I-protein
factor-1	NN	O	I-protein
(	NN	O	I-protein
IRF-1	NN	O	I-protein
)	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	O
NK	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

IL-12	NN	O	B-protein
is	NN	O	O
a	NN	O	O
critical	NN	O	B-protein
immunoregulatory	NN	O	I-protein
cytokine	NN	O	I-protein
that	NN	O	O
promotes	NN	O	O
cell-mediated	NN	O	O
immune	NN	O	O
responses	NN	O	O
and	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
naive	NN	O	O
CD4+	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
however	NN	O	O
,	NN	O	O
relatively	NN	O	O
few	NN	O	O
IL-12	NN	O	B-protein
target	NN	O	O
genes	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

To	NN	O	O
better	NN	O	O
clarify	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
action	NN	O	O
,	NN	O	O
we	NN	O	O
set	NN	O	O
out	NN	O	O
to	NN	O	O
characterize	NN	O	O
genes	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
first	NN	O	O
by	NN	O	O
contrasting	NN	O	O
IL-12	NN	O	B-DNA
-and	NN	O	O
IFN-alpha-inducible	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
identified	NN	O	O
several	NN	O	O
genes	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
namely	NN	O	O
,	NN	O	O
MIP-1alpha	NN	O	B-protein
,	NN	O	O
MIP-1beta	NN	O	B-protein
,	NN	O	O
IL-1RA	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IFN	NN	O	B-protein
regulatory	NN	O	I-protein
factor-1	NN	O	I-protein
(	NN	O	I-protein
IRF-1	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

IRF-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
regulated	NN	O	O
by	NN	O	O
IFNs	NN	O	B-protein
that	NN	O	O
is	NN	O	O
also	NN	O	O
essential	NN	O	O
for	NN	O	O
Th1	NN	O	B-cell_type
responses	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
IL-12	NN	O	B-protein
directly	NN	O	O
up-regulates	NN	O	O
IRF-1	NN	O	B-protein
to	NN	O	O
the	NN	O	O
same	NN	O	O
extent	NN	O	O
as	NN	O	O
IFN-alpha	NN	O	B-protein
in	NN	O	O
normal	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
showed	NN	O	O
that	NN	O	O
IL-12	NN	O	B-protein
had	NN	O	O
a	NN	O	O
direct	NN	O	O
effect	NN	O	O
on	NN	O	O
IRF-1	NN	O	B-protein
,	NN	O	O
an	NN	O	O
effect	NN	O	O
not	NN	O	O
mediated	NN	O	O
indirectly	NN	O	O
by	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
synergistically	NN	O	O
induced	NN	O	O
IRF-1	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
IFN-alpha	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
did	NN	O	O
not	NN	O	O
.	NN	O	O

The	NN	O	O
participation	NN	O	O
of	NN	O	O
STAT4	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
IRF-1	NN	O	B-protein
was	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
two	NN	O	O
ways	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
STAT4	NN	O	B-protein
was	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
IL-12	NN	O	B-protein
-dependent	NN	O	O
transactivation	NN	O	O
of	NN	O	O
an	NN	O	O
IRF-1	NN	O	B-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
second	NN	O	O
,	NN	O	O
STAT4	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
shown	NN	O	O
using	NN	O	O
EMSA	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
no	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
IRF-1	NN	O	B-protein
was	NN	O	O
found	NN	O	O
in	NN	O	O
IL-4-stimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
IL-4	NN	O	B-protein
did	NN	O	O
not	NN	O	O
block	NN	O	O
IL-12	NN	O	B-protein
-dependent	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
IRF-1	NN	O	B-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
IRF-1	NN	O	B-protein
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
contributor	NN	O	O
to	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
speculate	NN	O	O
that	NN	O	O
the	NN	O	O
defective	NN	O	O
IL-12	NN	O	B-protein
responses	NN	O	O
seen	NN	O	O
in	NN	O	O
IRF	NN	O	B-protein
-1-/-	NN	O	O
mice	NN	O	O
might	NN	O	O
be	NN	O	O
attributable	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
to	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Targeted	NN	O	O
remodeling	NN	O	O
of	NN	O	O
human	NN	O	O
beta-globin	NN	O	O
promoter	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
produces	NN	O	O
increased	NN	O	O
expression	NN	O	O
and	NN	O	O
decreased	NN	O	O
silencing	NN	O	O
.	NN	O	O

The	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
gene	NN	O	I-DNA
locus	NN	O	I-DNA
assumes	NN	O	O
a	NN	O	O
transcriptionally-active	NN	O	O
conformation	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

One	NN	O	O
feature	NN	O	O
of	NN	O	O
this	NN	O	O
chromatin	NN	O	O
reorganization	NN	O	O
is	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
DNase	NN	O	B-DNA
1	NN	O	I-DNA
hypersensitive	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
regions	NN	O	O
of	NN	O	O
active	NN	O	B-DNA
globin	NN	O	I-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
reorganization	NN	O	O
requires	NN	O	O
the	NN	O	O
globin	NN	O	B-DNA
locus	NN	O	I-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
normal	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
beta-like	NN	O	B-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
it	NN	O	O
is	NN	O	O
possible	NN	O	O
to	NN	O	O
artificially	NN	O	O
enhance	NN	O	O
the	NN	O	O
opening	NN	O	O
of	NN	O	O
the	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
of	NN	O	O
a	NN	O	O
minimal	NN	O	B-DNA
beta-globin	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
placed	NN	O	O
a	NN	O	O
101bp	NN	O	O
,	NN	O	O
erythroid-specific	NN	O	B-DNA
DNase	NN	O	I-DNA
1	NN	O	I-DNA
hypersensitive	NN	O	I-DNA
site-forming	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
HSFE	NN	O	B-DNA
)	NN	O	O
immediately	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
element	NN	O	O
includes	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
NF-E2	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
Sp-1	NN	O	B-protein
.	NN	O	O

Constructs	NN	O	O
were	NN	O	O
stably	NN	O	O
transfected	NN	O	O
into	NN	O	O
murine	NN	O	B-cell_type
erythroleukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
promoter	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
and	NN	O	O
gene	NN	O	O
expression	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

The	NN	O	O
HSFE	NN	O	B-DNA
induced	NN	O	O
an	NN	O	O
area	NN	O	O
of	NN	O	O
enhanced	NN	O	O
DNase	NN	O	B-protein
1	NN	O	I-protein
hypersensitivity	NN	O	O
extending	NN	O	O
from	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
start	NN	O	O
site	NN	O	O
to	NN	O	O
-300bp	NN	O	O
of	NN	O	O
the	NN	O	O
artificial	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
significantly	NN	O	O
increased	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
beta-globin	NN	O	B-DNA
promoters	NN	O	I-DNA
in	NN	O	O
an	NN	O	O
open	NN	O	O
chromatin	NN	O	O
configuration	NN	O	O
.	NN	O	O

This	NN	O	O
remodeling	NN	O	O
of	NN	O	O
promoter	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
resulted	NN	O	O
in	NN	O	O
3-fold	NN	O	O
increases	NN	O	O
in	NN	O	O
beta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
and	NN	O	O
induction	NN	O	O
,	NN	O	O
and	NN	O	O
inhibited	NN	O	O
long-term	NN	O	O
beta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
silencing	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
a	NN	O	O
relatively	NN	O	O
small	NN	O	O
cis-acting	NN	O	B-DNA
element	NN	O	I-DNA
is	NN	O	O
able	NN	O	O
to	NN	O	O
enhance	NN	O	O
remodeling	NN	O	O
of	NN	O	O
promoter	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
resulting	NN	O	O
in	NN	O	O
increased	NN	O	O
beta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
low	NN	O	O
shear	NN	O	O
flow-induced	NN	O	O
HAEC	NN	O	B-cell_line
VCAM-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
reported	NN	O	O
that	NN	O	O
prolonged	NN	O	O
exposure	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
aortic	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HAEC	NN	O	B-cell_line
)	NN	O	O
to	NN	O	O
low	NN	O	O
shear	NN	O	O
stress	NN	O	O
flow	NN	O	O
patterns	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
sustained	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
activated	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulator	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
investigate	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
low	NN	O	O
shear-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
resulting	NN	O	O
in	NN	O	O
augmented	NN	O	O
endothelial	NN	O	O
cell-monocyte	NN	O	O
(	NN	O	O
EC-Mn	NN	O	O
)	NN	O	O
adhesion	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
activation	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
intracellular	NN	O	O
oxidant	NN	O	O
activity	NN	O	O
.	NN	O	O

Before	NN	O	O
exposure	NN	O	O
to	NN	O	O
low	NN	O	O
shear	NN	O	O
(	NN	O	O
2	NN	O	O
dyn/cm2	NN	O	O
)	NN	O	O
for	NN	O	O
6	NN	O	O
h	NN	O	O
,	NN	O	O
HAEC	NN	O	B-cell_line
were	NN	O	O
preincubated	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
the	NN	O	O
antioxidants	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
or	NN	O	O
N-acetyl-L-cysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
.	NN	O	O

PDTC	NN	O	O
strongly	NN	O	O
inhibited	NN	O	O
low	NN	O	O
shear-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
expression	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
EC-Mn	NN	O	O
adhesion	NN	O	O
.	NN	O	O

Paradoxically	NN	O	O
,	NN	O	O
NAC	NN	O	O
exerted	NN	O	O
a	NN	O	O
positive	NN	O	O
effect	NN	O	O
on	NN	O	O
low	NN	O	O
shear-induced	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
EC-Mn	NN	O	O
adhesion	NN	O	O
and	NN	O	O
only	NN	O	O
slightly	NN	O	O
downregulated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
cytokine-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
are	NN	O	O
blocked	NN	O	O
by	NN	O	O
both	NN	O	O
PDTC	NN	O	O
and	NN	O	O
NAC	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
low	NN	O	O
shear-induced	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
that	NN	O	O
pathways	NN	O	O
mediating	NN	O	O
low	NN	O	O
shear-	NN	O	O
and	NN	O	O
cytokine-induced	NN	O	O
EC-Mn	NN	O	O
adhesion	NN	O	O
may	NN	O	O
be	NN	O	O
differentially	NN	O	O
regulated	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor-90	NN	O	I-protein
of	NN	O	O
activated	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
:	NN	O	O
A	NN	O	O
double-stranded	NN	O	B-RNA
RNA	NN	O	I-RNA
-binding	NN	O	O
protein	NN	O	O
and	NN	O	O
substrate	NN	O	O
for	NN	O	O
the	NN	O	O
double-stranded	NN	O	B-protein
RNA-dependent	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
PKR	NN	O	B-protein
.	NN	O	O

NFAT	NN	O	O
transcription	NN	O	O
factors	NN	O	O
play	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
initiating	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
through	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
immediate-early	NN	O	O
T-cell	NN	O	O
specific	NN	O	O
genes	NN	O	O
including	NN	O	O
interleukin-2	NN	O	O
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
.	NN	O	O

NFAT	NN	O	O
transcription	NN	O	O
factors	NN	O	O
bind	NN	O	O
to	NN	O	O
a	NN	O	O
sequence	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
known	NN	O	O
as	NN	O	O
the	NN	O	O
antigen	NN	O	B-DNA
receptor	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
2	NN	O	I-DNA
(	NN	O	O
ARRE-2	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Multiple	NN	O	O
proteins	NN	O	O
exhibiting	NN	O	O
ARRE-2	NN	O	B-DNA
binding	NN	O	O
activity	NN	O	O
have	NN	O	O
been	NN	O	O
isolated	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
heterodimer	NN	O	B-protein
from	NN	O	O
stimulated	NN	O	O
T-cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
consisting	NN	O	O
of	NN	O	O
Mr	NN	O	O
=	NN	O	O
90	NN	O	O
000	NN	O	O
(	NN	O	O
NF90	NN	O	B-protein
)	NN	O	O
and	NN	O	O
Mr	NN	O	O
=	NN	O	O
45	NN	O	O
000	NN	O	O
(	NN	O	O
NF45	NN	O	B-protein
)	NN	O	O
subunits	NN	O	O
.	NN	O	O

The	NN	O	O
subunits	NN	O	O
of	NN	O	O
this	NN	O	O
heterodimer	NN	O	B-protein
have	NN	O	O
been	NN	O	O
cloned	NN	O	O
,	NN	O	O
and	NN	O	O
NF90	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
encode	NN	O	O
a	NN	O	O
protein	NN	O	O
containing	NN	O	O
two	NN	O	O
domains	NN	O	O
that	NN	O	O
are	NN	O	O
predicted	NN	O	O
to	NN	O	O
form	NN	O	O
motifs	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
double-stranded	NN	O	B-RNA
RNA	NN	O	I-RNA
.	NN	O	O

Using	NN	O	O
in	NN	O	O
vitro	NN	O	O
translated	NN	O	O
polypeptides	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
NF90	NN	O	B-protein
specifically	NN	O	O
binds	NN	O	O
to	NN	O	O
double-stranded	NN	O	B-RNA
RNA	NN	O	I-RNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
NF90	NN	O	B-protein
was	NN	O	O
phosphorylated	NN	O	O
in	NN	O	O
a	NN	O	O
double-stranded	NN	O	B-RNA
RNA	NN	O	I-RNA
-dependent	NN	O	O
manner	NN	O	O
likely	NN	O	O
by	NN	O	O
the	NN	O	O
interferon-induced	NN	O	O
,	NN	O	O
double-stranded	NN	O	B-protein
RNA-dependent	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
PKR	NN	O	B-protein
.	NN	O	O

The	NN	O	O
NF90	NN	O	B-protein
protein	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
not	NN	O	O
only	NN	O	O
in	NN	O	O
T-cells	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
nonimmune	NN	O	B-cell_line
HeLa	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
protein	NN	O	O
was	NN	O	O
almost	NN	O	O
exclusively	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
ribosome	NN	O	O
salt	NN	O	O
wash	NN	O	O
fraction	NN	O	O
of	NN	O	O
cell	NN	O	O
lysates	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappaB	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
C5a-induced	NN	O	B-DNA
interleukin-8	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

C5a	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
peptide	NN	O	O
chemoattractant	NN	O	O
,	NN	O	O
stimulates	NN	O	O
interleukin-8	NN	O	B-protein
(	NN	O	O
IL-8	NN	O	B-protein
)	NN	O	O
secretion	NN	O	O
from	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

Experiments	NN	O	O
were	NN	O	O
conducted	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
C5a-induced	NN	O	O
IL-8	NN	O	B-protein
production	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
14-fold	NN	O	O
greater	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
unstimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
2	NN	O	O
hours	NN	O	O
.	NN	O	O

IL-8	NN	O	B-protein
secretion	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
accumulation	NN	O	O
of	NN	O	O
IL-8	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
and	NN	O	O
by	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
within	NN	O	O
30	NN	O	O
minutes	NN	O	O
.	NN	O	O

AP-1	NN	O	B-protein
but	NN	O	O
not	NN	O	O
NF-IL-6	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
also	NN	O	O
detected	NN	O	O
in	NN	O	O
C5a-stimulated	NN	O	B-cell_line
PBMC	NN	O	I-cell_line
;	NN	O	O
however	NN	O	O
,	NN	O	O
its	NN	O	O
delayed	NN	O	O
expression	NN	O	O
(	NN	O	O
maximal	NN	O	O
at	NN	O	O
4	NN	O	O
hours	NN	O	O
)	NN	O	O
suggested	NN	O	O
a	NN	O	O
less	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
rapid	NN	O	O
production	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
.	NN	O	O

The	NN	O	O
correlation	NN	O	O
between	NN	O	O
C5a-induced	NN	O	O
kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
IL-8	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
the	NN	O	O
RAW264.7	NN	O	B-cell_line
macrophage	NN	O	I-cell_line
cells	NN	O	I-cell_line
using	NN	O	O
reporter	NN	O	O
genes	NN	O	O
directed	NN	O	O
by	NN	O	O
the	NN	O	O
kappaB	NN	O	B-DNA
sequence	NN	O	I-DNA
from	NN	O	O
IkappaBalpha	NN	O	B-protein
and	NN	O	O
IL-8	NN	O	B-DNA
promoter	NN	O	I-DNA
regions	NN	O	I-DNA
.	NN	O	O

C5a-induced	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
abolished	NN	O	O
by	NN	O	O
introducing	NN	O	O
mutations	NN	O	O
into	NN	O	O
the	NN	O	O
kappaB	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
by	NN	O	O
coexpression	NN	O	O
of	NN	O	O
a	NN	O	O
dominant	NN	O	B-DNA
negative	NN	O	I-DNA
IkappaBalpha	NN	O	I-DNA
construct	NN	O	I-DNA
resistant	NN	O	O
to	NN	O	O
agonist-induced	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

Pertussis	NN	O	O
toxin	NN	O	O
,	NN	O	O
which	NN	O	O
ADP-ribosylates	NN	O	O
the	NN	O	O
Gi	NN	O	B-protein
proteins	NN	O	I-protein
known	NN	O	O
to	NN	O	O
couple	NN	O	O
to	NN	O	O
the	NN	O	O
C5a	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
produced	NN	O	O
minimal	NN	O	O
inhibition	NN	O	O
of	NN	O	O
C5a-induced	NN	O	O
IL-8	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
had	NN	O	O
little	NN	O	O
effect	NN	O	O
on	NN	O	O
C5a-induced	NN	O	O
calcium	NN	O	O
mobilization	NN	O	O
in	NN	O	O
RAW264.7	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
C5a-induced	NN	O	O
IL-8	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
response	NN	O	O
is	NN	O	O
mediated	NN	O	O
primarily	NN	O	O
through	NN	O	O
a	NN	O	O
pertussis	NN	O	O
toxin-insensitive	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Increased	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
beta	NN	O	I-protein
in	NN	O	O
airway	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
glucocorticoid-insensitive	NN	O	O
asthma	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
-insensitive	NN	O	O
asthma	NN	O	O
is	NN	O	O
a	NN	O	O
challenging	NN	O	O
clinical	NN	O	O
problem	NN	O	O
that	NN	O	O
can	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
life-threatening	NN	O	O
disease	NN	O	O
progression	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
GC	NN	O	O
insensitivity	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

Alternative	NN	O	O
splicing	NN	O	O
of	NN	O	O
the	NN	O	O
GC	NN	O	B-RNA
receptor	NN	O	I-RNA
(	NN	O	I-RNA
GCR	NN	O	I-RNA
)	NN	O	I-RNA
pre-mRNA	NN	O	I-RNA
generates	NN	O	O
a	NN	O	O
second	NN	O	O
GCR	NN	O	B-protein
,	NN	O	O
termed	NN	O	O
GCRbeta	NN	O	B-protein
,	NN	O	O
which	NN	O	O
does	NN	O	O
not	NN	O	O
bind	NN	O	O
GC	NN	O	O
but	NN	O	O
antagonizes	NN	O	O
the	NN	O	O
transactivating	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
classic	NN	O	O
GCR	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
GCRbeta	NN	O	B-protein
could	NN	O	O
account	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	O
insensitivity	NN	O	O
.	NN	O	O

Bronchoalveolar	NN	O	B-cell_type
lavage	NN	O	I-cell_type
(	NN	O	I-cell_type
BAL	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
GCRbeta	NN	O	B-protein
immunoreactivity	NN	O	O
using	NN	O	O
a	NN	O	O
GCRbeta-specific	NN	O	B-protein
antibody	NN	O	I-protein
by	NN	O	O
immunohistochemical	NN	O	O
staining	NN	O	O
.	NN	O	O

Cell	NN	O	O
localization	NN	O	O
of	NN	O	O
GCRbeta	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
performed	NN	O	O
using	NN	O	O
a	NN	O	O
double	NN	O	O
immunostaining	NN	O	O
technique	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
GC-insensitive	NN	O	O
asthma	NN	O	O
expressed	NN	O	O
a	NN	O	O
significantly	NN	O	O
higher	NN	O	O
number	NN	O	O
of	NN	O	O
GCRbeta-immunoreactive	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
their	NN	O	O
BAL	NN	O	O
and	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
than	NN	O	O
GC-sensitive	NN	O	O
asthmatics	NN	O	O
or	NN	O	O
normal	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
GCRbeta	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
GC-insensitive	NN	O	O
asthma	NN	O	O
was	NN	O	O
particularly	NN	O	O
high	NN	O	O
in	NN	O	O
airway	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
asthma	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
examined	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
GCRbeta	NN	O	B-protein
in	NN	O	O
specimens	NN	O	O
from	NN	O	O
the	NN	O	O
airways	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
bronchitis	NN	O	O
.	NN	O	O

In	NN	O	O
chronic	NN	O	O
bronchitis	NN	O	O
,	NN	O	O
few	NN	O	O
cells	NN	O	O
were	NN	O	O
GCRbeta-positive	NN	O	B-cell_type
and	NN	O	O
their	NN	O	O
numbers	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
significantly	NN	O	O
from	NN	O	O
normal	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
GC-insensitive	NN	O	O
asthma	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
GCRbeta	NN	O	B-protein
in	NN	O	O
airway	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
-STAT5a	NN	O	B-protein
pathway	NN	O	O
after	NN	O	O
CD40	NN	O	B-protein
triggering	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CD40	NN	O	B-protein
/CD40	NN	O	B-protein
ligand	NN	O	O
interactions	NN	O	O
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
responses	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
triggered	NN	O	O
by	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
receptor	NN	O	I-protein
have	NN	O	O
been	NN	O	O
widely	NN	O	O
studied	NN	O	O
in	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
signaling	NN	O	O
following	NN	O	O
CD40	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
resting	NN	O	B-cell_line
tonsillar	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
pathway	NN	O	O
in	NN	O	O
highly	NN	O	O
purified	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
resting	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

After	NN	O	O
CD40	NN	O	B-protein
triggering	NN	O	O
,	NN	O	O
a	NN	O	O
similar	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
(	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
complex	NN	O	I-protein
occurred	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
preparations	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
complexes	NN	O	I-protein
were	NN	O	O
different	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
B	NN	O	O
cells	NN	O	O
,	NN	O	O
because	NN	O	O
p50	NN	O	B-protein
is	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
complex	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
triggering	NN	O	O
in	NN	O	O
both	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
p65	NN	O	B-protein
was	NN	O	O
only	NN	O	O
induced	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O
In	NN	O	O
contrast	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
was	NN	O	O
associated	NN	O	O
with	NN	O	O
CD40	NN	O	B-protein
molecules	NN	O	O
in	NN	O	O
both	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
resting	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
phosphorylation	NN	O	O
induction	NN	O	O
was	NN	O	O
observed	NN	O	O
only	NN	O	O
in	NN	O	O
CD40-activated	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
,	NN	O	O
with	NN	O	O
subsequent	NN	O	O
induction	NN	O	O
of	NN	O	O
STAT5a	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
signals	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
differ	NN	O	O
following	NN	O	O
CD40	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
normal	NN	O	O
CD40	NN	O	B-protein
-induced	NN	O	O
monocyte	NN	O	O
activation	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
CD40	NN	O	B-protein
ligand+	NN	O	O
hyper	NN	O	O
IgM	NN	O	B-protein
syndrome	NN	O	O
in	NN	O	O
whom	NN	O	O
a	NN	O	O
defect	NN	O	O
in	NN	O	O
CD40	NN	O	B-protein
-induced	NN	O	O
B	NN	O	O
cell	NN	O	O
activation	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
.	NN	O	O

-DOCSTART-	O

Genetic	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
additional	NN	O	O
factor	NN	O	O
required	NN	O	O
for	NN	O	O
erythropoietin	NN	O	B-protein
-induced	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

Erythropoietin	NN	O	B-protein
(	NN	O	O
EPO	NN	O	B-protein
)	NN	O	O
and	NN	O	O
its	NN	O	O
receptor	NN	O	O
(	NN	O	O
EPOR	NN	O	B-protein
)	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
mature	NN	O	O
erythrocytes	NN	O	O
.	NN	O	O

After	NN	O	O
binding	NN	O	O
of	NN	O	O
ligand	NN	O	O
,	NN	O	O
the	NN	O	O
EPOR	NN	O	B-protein
activates	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
that	NN	O	O
ultimately	NN	O	O
control	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
,	NN	O	O
survival	NN	O	O
,	NN	O	O
and	NN	O	O
specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Although	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
appear	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
principal	NN	O	O
EPO-responsive	NN	O	B-cell_type
cell	NN	O	I-cell_type
type	NN	O	I-cell_type
in	NN	O	O
vivo	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
restricted	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
EPOR	NN	O	B-protein
,	NN	O	O
many	NN	O	O
growth	NN	O	B-cell_line
factor-dependent	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
expressing	NN	O	O
the	NN	O	O
EPOR	NN	O	B-protein
can	NN	O	O
respond	NN	O	O
to	NN	O	O
EPO	NN	O	B-protein
by	NN	O	O
activating	NN	O	O
many	NN	O	O
or	NN	O	O
all	NN	O	O
of	NN	O	O
these	NN	O	O
pathways	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
cellular	NN	O	O
context	NN	O	O
(	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-cell_line
[	NN	O	I-cell_line
IL-2	NN	O	I-cell_line
]	NN	O	I-cell_line
-dependent	NN	O	I-cell_line
HT-2	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
EPO	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
EPOR	NN	O	B-protein
fails	NN	O	O
to	NN	O	O
support	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
,	NN	O	O
STAT-5	NN	O	B-protein
induction	NN	O	O
,	NN	O	O
or	NN	O	O
MAPK	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
despite	NN	O	O
efficient	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
EPOR	NN	O	B-protein
and	NN	O	O
JAK2	NN	O	B-protein
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
after	NN	O	O
withdrawal	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
when	NN	O	O
we	NN	O	O
fused	NN	O	O
HT-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
the	NN	O	O
EPOR	NN	O	B-protein
with	NN	O	O
Ba/F3	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
a	NN	O	O
complementation	NN	O	O
assay	NN	O	O
,	NN	O	O
the	NN	O	O
resulting	NN	O	O
hybridomas	NN	O	B-cell_line
proliferated	NN	O	O
and	NN	O	O
potently	NN	O	O
activated	NN	O	O
STAT-5	NN	O	B-protein
and	NN	O	O
MAPK	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
EPO	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
an	NN	O	O
unidentified	NN	O	B-protein
cellular	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
needed	NN	O	O
to	NN	O	O
mediate	NN	O	O
signaling	NN	O	O
by	NN	O	O
the	NN	O	O
EPOR	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
Ba/F3	NN	O	B-cell_line
cells	NN	O	I-cell_line
apparently	NN	O	O
express	NN	O	O
this	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
and	NN	O	O
somatic	NN	O	O
fusions	NN	O	O
can	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
confer	NN	O	O
EPO	NN	O	B-protein
-responsiveness	NN	O	O
to	NN	O	O
HT-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
lack	NN	O	O
this	NN	O	O
factor	NN	O	O
.	NN	O	O

-DOCSTART-	O

Novel	NN	O	O
therapies	NN	O	O
for	NN	O	O
inflammatory	NN	O	O
bowel	NN	O	O
disease	NN	O	O
.	NN	O	O

Looking	NN	O	O
back	NN	O	O
at	NN	O	O
successes	NN	O	O
and	NN	O	O
failures	NN	O	O
in	NN	O	O
newer	NN	O	O
approaches	NN	O	O
to	NN	O	O
treating	NN	O	O
IBD	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
tempting	NN	O	O
--	NN	O	O
although	NN	O	O
still	NN	O	O
difficult	NN	O	O
--	NN	O	O
to	NN	O	O
draw	NN	O	O
conclusions	NN	O	O
about	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

When	NN	O	O
a	NN	O	O
therapy	NN	O	O
proves	NN	O	O
effective	NN	O	O
,	NN	O	O
do	NN	O	O
clinicians	NN	O	O
truly	NN	O	O
know	NN	O	O
how	NN	O	O
it	NN	O	O
works	NN	O	O
?	NN	O	O
Even	NN	O	O
with	NN	O	O
a	NN	O	O
therapy	NN	O	O
as	NN	O	O
specific	NN	O	O
as	NN	O	O
anti-TNF	NN	O	B-protein
antibody	NN	O	I-protein
,	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
clear	NN	O	O
if	NN	O	O
the	NN	O	O
benefit	NN	O	O
is	NN	O	O
attributable	NN	O	O
to	NN	O	O
simple	NN	O	O
binding	NN	O	O
and	NN	O	O
clearance	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
or	NN	O	O
to	NN	O	O
binding	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
and	NN	O	O
subsequent	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
activated	NN	O	B-cell_type
macrophage	NN	O	I-cell_type
.	NN	O	O

When	NN	O	O
a	NN	O	O
drug	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
less	NN	O	O
effective	NN	O	O
than	NN	O	O
preclinical	NN	O	O
models	NN	O	O
suggest	NN	O	O
,	NN	O	O
can	NN	O	O
failures	NN	O	O
in	NN	O	O
effectiveness	NN	O	O
from	NN	O	O
delivery	NN	O	O
or	NN	O	O
dosing	NN	O	O
be	NN	O	O
differentiated	NN	O	O
?	NN	O	O
The	NN	O	O
disappointing	NN	O	O
results	NN	O	O
of	NN	O	O
clinical	NN	O	O
trials	NN	O	O
with	NN	O	O
IL-10	NN	O	B-protein
--	NN	O	O
so	NN	O	O
at	NN	O	O
odds	NN	O	O
with	NN	O	O
the	NN	O	O
prediction	NN	O	O
of	NN	O	O
benefit	NN	O	O
from	NN	O	O
animal	NN	O	O
models	NN	O	O
--	NN	O	O
bring	NN	O	O
into	NN	O	O
question	NN	O	O
the	NN	O	O
validity	NN	O	O
of	NN	O	O
those	NN	O	O
models	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
soundness	NN	O	O
of	NN	O	O
design	NN	O	O
of	NN	O	O
the	NN	O	O
clinical	NN	O	O
trials	NN	O	O
on	NN	O	O
which	NN	O	O
efficacy	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
is	NN	O	O
judged	NN	O	O
.	NN	O	O

The	NN	O	O
variability	NN	O	O
of	NN	O	O
response	NN	O	O
even	NN	O	O
to	NN	O	O
the	NN	O	O
most	NN	O	O
narrowly	NN	O	O
targeted	NN	O	O
agents	NN	O	O
suggests	NN	O	O
that	NN	O	O
these	NN	O	O
diseases	NN	O	O
are	NN	O	O
far	NN	O	O
more	NN	O	O
heterogeneous	NN	O	O
in	NN	O	O
humans	NN	O	O
than	NN	O	O
in	NN	O	O
their	NN	O	O
murine	NN	O	O
counterparts	NN	O	O
.	NN	O	O

Clinicians	NN	O	O
are	NN	O	O
only	NN	O	O
just	NN	O	O
beginning	NN	O	O
to	NN	O	O
recognize	NN	O	O
subclinical	NN	O	O
markers	NN	O	O
of	NN	O	O
response	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
may	NN	O	O
soon	NN	O	O
be	NN	O	O
possible	NN	O	O
to	NN	O	O
predict	NN	O	O
response	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
genetic	NN	O	O
composition	NN	O	O
.	NN	O	O

For	NN	O	O
the	NN	O	O
moment	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
field	NN	O	O
of	NN	O	O
pharmacogenetics	NN	O	O
is	NN	O	O
embryonic	NN	O	O
.	NN	O	O

Challenges	NN	O	O
in	NN	O	O
developing	NN	O	O
new	NN	O	O
therapeutic	NN	O	O
strategies	NN	O	O
include	NN	O	O
not	NN	O	O
only	NN	O	O
identifying	NN	O	O
novel	NN	O	O
agents	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
improving	NN	O	O
the	NN	O	O
definitions	NN	O	O
of	NN	O	O
clinical	NN	O	O
endpoints	NN	O	O
and	NN	O	O
defining	NN	O	O
efficacy	NN	O	O
at	NN	O	O
the	NN	O	O
biologic	NN	O	O
level	NN	O	O
.	NN	O	O

Only	NN	O	O
through	NN	O	O
considered	NN	O	O
evaluation	NN	O	O
of	NN	O	O
clinical	NN	O	O
evidence	NN	O	O
may	NN	O	O
clinicians	NN	O	O
determine	NN	O	O
which	NN	O	O
therapies	NN	O	O
should	NN	O	O
remain	NN	O	O
novelties	NN	O	O
and	NN	O	O
which	NN	O	O
should	NN	O	O
become	NN	O	O
an	NN	O	O
accepted	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
armamentarium	NN	O	O
.	NN	O	O

-DOCSTART-	O

Potent	NN	O	O
and	NN	O	O
stable	NN	O	O
attenuation	NN	O	O
of	NN	O	O
live-HIV-1	NN	O	O
by	NN	O	O
gain	NN	O	O
of	NN	O	O
a	NN	O	O
proteolysis-resistant	NN	O	B-protein
inhibitor	NN	O	I-protein
of	NN	O	O
NF-kappaB	NN	O	B-protein
(	NN	O	O
IkappaB-alphaS32/36A	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
implications	NN	O	O
for	NN	O	O
vaccine	NN	O	O
development	NN	O	O
.	NN	O	O

Live-attenuated	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
viruses	NN	O	O
(	NN	O	O
HIVs	NN	O	O
)	NN	O	O
are	NN	O	O
candidates	NN	O	O
for	NN	O	O
Acquired	NN	O	O
Immunodeficiency	NN	O	O
Syndrome	NN	O	O
(	NN	O	O
AIDS	NN	O	O
)	NN	O	O
vaccine	NN	O	O
.	NN	O	O

Based	NN	O	O
on	NN	O	O
the	NN	O	O
simian	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
SIV	NN	O	O
)	NN	O	O
model	NN	O	O
for	NN	O	O
AIDS	NN	O	O
,	NN	O	O
loss-of-function	NN	O	O
(	NN	O	O
e.g.	NN	O	O
deletion	NN	O	O
of	NN	O	O
accessory	NN	O	O
genes	NN	O	O
such	NN	O	O
as	NN	O	O
nef	NN	O	B-DNA
)	NN	O	O
has	NN	O	O
been	NN	O	O
forwarded	NN	O	O
as	NN	O	O
a	NN	O	O
primary	NN	O	O
approach	NN	O	O
for	NN	O	O
creating	NN	O	O
enfeebled	NN	O	O
,	NN	O	O
but	NN	O	O
replication-competent	NN	O	O
,	NN	O	O
HIV-1/SIV	NN	O	O
.	NN	O	O

Regrettably	NN	O	O
,	NN	O	O
recent	NN	O	O
evidence	NN	O	O
suggests	NN	O	O
that	NN	O	O
loss-of-function	NN	O	O
alone	NN	O	O
is	NN	O	O
not	NN	O	O
always	NN	O	O
sufficient	NN	O	O
to	NN	O	O
prevent	NN	O	O
the	NN	O	O
emergence	NN	O	O
of	NN	O	O
virulent	NN	O	O
mutants	NN	O	O
.	NN	O	O

New	NN	O	O
strategies	NN	O	O
that	NN	O	O
attenuate	NN	O	O
via	NN	O	O
mechanisms	NN	O	O
distinct	NN	O	O
from	NN	O	O
loss-of-function	NN	O	O
are	NN	O	O
needed	NN	O	O
for	NN	O	O
enhancing	NN	O	O
the	NN	O	O
safety	NN	O	O
phenotype	NN	O	O
of	NN	O	O
viral	NN	O	B-DNA
genome	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
gain-of-function	NN	O	O
to	NN	O	O
be	NN	O	O
used	NN	O	O
simultaneously	NN	O	O
with	NN	O	O
loss-of-function	NN	O	O
as	NN	O	O
a	NN	O	O
novel	NN	O	O
approach	NN	O	O
for	NN	O	O
attenuating	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
constructed	NN	O	O
an	NN	O	O
HIV-1	NN	O	B-DNA
genome	NN	O	I-DNA
carrying	NN	O	O
the	NN	O	O
cDNA	NN	O	B-DNA
of	NN	O	O
a	NN	O	O
proteolysis-resistant	NN	O	B-protein
nuclear	NN	O	I-protein
factor-kappaB	NN	O	I-protein
inhibitor	NN	O	I-protein
(	NN	O	O
IkappaB-alphaS32/36A	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
nef	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

HIV-1	NN	O	O
expressing	NN	O	O
IkappaB-alphaS32/36A	NN	O	B-protein
down-regulates	NN	O	O
viral	NN	O	O
expression	NN	O	O
and	NN	O	O
is	NN	O	O
highly	NN	O	O
attenuated	NN	O	O
in	NN	O	O
both	NN	O	O
Jurkat	NN	O	B-cell_line
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
provide	NN	O	O
formal	NN	O	O
proof	NN	O	O
that	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
and	NN	O	O
attenuating	NN	O	O
characteristics	NN	O	O
of	NN	O	O
IkappaB-alphaS32/36A	NN	O	B-protein
permit	NN	O	O
its	NN	O	O
stable	NN	O	O
maintenance	NN	O	O
in	NN	O	O
a	NN	O	O
live	NN	O	O
,	NN	O	O
replicating	NN	O	O
HIV-1	NN	O	O
despite	NN	O	O
180	NN	O	O
days	NN	O	O
of	NN	O	O
forced	NN	O	O
ex	NN	O	O
vivo	NN	O	O
passaging	NN	O	O
in	NN	O	O
tissue	NN	O	O
culture	NN	O	O
.	NN	O	O

As	NN	O	O
compared	NN	O	O
with	NN	O	O
other	NN	O	O
open-reading	NN	O	B-DNA
frames	NN	O	I-DNA
embedded	NN	O	O
into	NN	O	O
HIV/SIV	NN	O	B-DNA
genome	NN	O	I-DNA
,	NN	O	O
this	NN	O	O
degree	NN	O	O
of	NN	O	O
stability	NN	O	O
is	NN	O	O
unprecedented	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
IkappaB-alphaS32/36A	NN	O	B-protein
offers	NN	O	O
proof-of-principle	NN	O	O
that	NN	O	O
artifactually	NN	O	O
gained	NN	O	O
functions	NN	O	O
,	NN	O	O
when	NN	O	O
used	NN	O	O
to	NN	O	O
attenuate	NN	O	O
the	NN	O	O
replication	NN	O	O
of	NN	O	O
live	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
can	NN	O	O
be	NN	O	O
stable	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
illustrate	NN	O	O
gain-of-function	NN	O	O
as	NN	O	O
a	NN	O	O
feasible	NN	O	O
strategy	NN	O	O
for	NN	O	O
developing	NN	O	O
safer	NN	O	O
live-attenuated	NN	O	O
HIVs	NN	O	O
to	NN	O	O
be	NN	O	O
tested	NN	O	O
as	NN	O	O
candidates	NN	O	O
for	NN	O	O
AIDS	NN	O	O
vaccine	NN	O	O
.	NN	O	O

-DOCSTART-	O

Multiple	NN	O	O
NF-ATc	NN	O	B-protein
isoforms	NN	O	I-protein
with	NN	O	O
individual	NN	O	O
transcriptional	NN	O	O
properties	NN	O	O
are	NN	O	O
synthesized	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-ATc	NN	O	B-protein
that	NN	O	O
controls	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
embryonic	NN	O	B-cell_type
cardiac	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
three	NN	O	O
prominent	NN	O	O
isoforms	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
alternative	NN	O	O
splice/polyadenylation	NN	O	O
events	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
predominant	NN	O	O
synthesis	NN	O	O
of	NN	O	O
two	NN	O	O
long	NN	O	O
isoforms	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
a	NN	O	O
shorter	NN	O	O
NF-ATc	NN	O	B-protein
isoform	NN	O	I-protein
in	NN	O	O
effector	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Whereas	NN	O	O
the	NN	O	O
previously	NN	O	O
described	NN	O	O
isoform	NN	O	B-protein
NF-ATc/A	NN	O	I-protein
contains	NN	O	O
a	NN	O	O
relatively	NN	O	O
short	NN	O	O
C	NN	O	B-protein
terminus	NN	O	I-protein
,	NN	O	O
the	NN	O	O
longer	NN	O	O
isoforms	NN	O	O
,	NN	O	O
B	NN	O	B-protein
and	NN	O	O
C	NN	O	B-protein
,	NN	O	O
span	NN	O	O
extra	NN	O	O
C-terminal	NN	O	O
peptides	NN	O	O
of	NN	O	O
128	NN	O	O
and	NN	O	O
246	NN	O	O
aa	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
strong	NN	O	B-protein
N-terminal	NN	O	I-protein
trans-activation	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
TAD-A	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
common	NN	O	O
to	NN	O	O
all	NN	O	O
three	NN	O	O
NF-ATc	NN	O	B-protein
isoforms	NN	O	I-protein
,	NN	O	O
NF-ATc/C	NN	O	B-protein
contains	NN	O	O
a	NN	O	O
second	NN	O	O
trans-activation	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
TAD-B	NN	O	B-protein
,	NN	O	O
in	NN	O	O
its	NN	O	O
C-terminal	NN	O	O
peptide	NN	O	O
.	NN	O	O

Various	NN	O	O
stimuli	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
induce	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
TAD-A	NN	O	B-protein
also	NN	O	O
enhance	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
TAD-B	NN	O	B-protein
,	NN	O	O
but	NN	O	O
,	NN	O	O
unlike	NN	O	O
TAD-A	NN	O	B-protein
,	NN	O	O
TAD-B	NN	O	B-protein
remains	NN	O	O
unphosphorylated	NN	O	O
by	NN	O	O
protein	NN	O	O
from	NN	O	O
12-O-tetradecanoyl	NN	O	B-cell_line
12-phorbol	NN	O	I-cell_line
13-acetate-stimulated	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
shorter	NN	O	O
C-terminal	NN	O	O
peptide	NN	O	O
of	NN	O	O
isoform	NN	O	O
NF-ATc/B	NN	O	B-protein
exerts	NN	O	O
a	NN	O	O
suppressive	NN	O	O
transcriptional	NN	O	O
effect	NN	O	O
.	NN	O	O

These	NN	O	O
properties	NN	O	O
of	NN	O	O
NF-ATc/B	NN	O	B-protein
and	NN	O	I-protein
-C	NN	O	I-protein
might	NN	O	O
be	NN	O	O
of	NN	O	O
importance	NN	O	O
for	NN	O	O
gene	NN	O	O
regulation	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
which	NN	O	O
NF-ATc/B	NN	O	B-protein
and	NN	O	I-protein
-C	NN	O	I-protein
are	NN	O	O
predominantly	NN	O	O
synthesized	NN	O	O
.	NN	O	O

-DOCSTART-	O

Selection	NN	O	O
and	NN	O	O
long-term	NN	O	O
persistence	NN	O	O
of	NN	O	O
reactive	NN	O	O
CTL	NN	O	B-cell_line
clones	NN	O	I-cell_line
during	NN	O	O
an	NN	O	O
EBV	NN	O	O
chronic	NN	O	O
response	NN	O	O
are	NN	O	O
determined	NN	O	O
by	NN	O	O
avidity	NN	O	O
,	NN	O	O
CD8	NN	O	B-protein
variable	NN	O	O
contribution	NN	O	O
compensating	NN	O	O
for	NN	O	O
differences	NN	O	O
in	NN	O	O
TCR	NN	O	B-protein
affinities	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
diversity	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
repertoire	NN	O	O
after	NN	O	O
primary	NN	O	O
immunization	NN	O	O
is	NN	O	O
conserved	NN	O	O
in	NN	O	O
memory	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
a	NN	O	O
progressive	NN	O	O
narrowing	NN	O	O
of	NN	O	O
this	NN	O	O
repertoire	NN	O	O
may	NN	O	O
take	NN	O	O
place	NN	O	O
during	NN	O	O
recall	NN	O	O
infections	NN	O	O
.	NN	O	O

It	NN	O	O
now	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
investigated	NN	O	O
which	NN	O	O
parameters	NN	O	O
determine	NN	O	O
the	NN	O	O
repertoire	NN	O	O
of	NN	O	O
the	NN	O	O
memory	NN	O	O
response	NN	O	O
and	NN	O	O
possibly	NN	O	O
restrict	NN	O	O
its	NN	O	O
diversity	NN	O	O
after	NN	O	O
subsequent	NN	O	O
antigenic	NN	O	O
challenges	NN	O	O
.	NN	O	O

To	NN	O	O
address	NN	O	O
this	NN	O	O
question	NN	O	O
,	NN	O	O
we	NN	O	O
took	NN	O	O
advantage	NN	O	O
of	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
CD8+	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
joint	NN	O	O
of	NN	O	O
a	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
patient	NN	O	O
and	NN	O	O
selected	NN	O	O
for	NN	O	O
their	NN	O	O
reactivity	NN	O	O
against	NN	O	O
a	NN	O	O
single	NN	O	B-protein
MHC/peptide	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
both	NN	O	O
TCR	NN	O	B-protein
chains	NN	O	I-protein
documented	NN	O	O
a	NN	O	O
great	NN	O	O
diversity	NN	O	O
among	NN	O	O
those	NN	O	O
clones	NN	O	O
and	NN	O	O
the	NN	O	O
persistence	NN	O	O
of	NN	O	O
clonotypes	NN	O	O
over	NN	O	O
a	NN	O	O
2-yr	NN	O	O
period	NN	O	O
.	NN	O	O

Strikingly	NN	O	O
,	NN	O	O
despite	NN	O	O
the	NN	O	O
observed	NN	O	O
repertoire	NN	O	O
heterogeneity	NN	O	O
,	NN	O	O
all	NN	O	O
clones	NN	O	O
displayed	NN	O	O
a	NN	O	O
narrow	NN	O	O
range	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
/peptide	NN	O	O
density	NN	O	O
requirements	NN	O	O
in	NN	O	O
cytotoxicity	NN	O	O
assays	NN	O	O
(	NN	O	O
ED50	NN	O	O
between	NN	O	O
9	NN	O	O
and	NN	O	O
36	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

TCR	NN	O	B-protein
affinities	NN	O	O
were	NN	O	O
then	NN	O	O
indirectly	NN	O	O
estimated	NN	O	O
by	NN	O	O
blocking	NN	O	O
CD8	NN	O	B-protein
interaction	NN	O	O
with	NN	O	O
an	NN	O	O
anti-CD8	NN	O	B-protein
mAb	NN	O	I-protein
.	NN	O	O

We	NN	O	O
found	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
affinities	NN	O	O
among	NN	O	O
the	NN	O	O
different	NN	O	O
clonotypes	NN	O	O
that	NN	O	O
segregated	NN	O	O
with	NN	O	O
Vbeta	NN	O	O
usage	NN	O	O
.	NN	O	O

We	NN	O	O
thus	NN	O	O
propose	NN	O	O
that	NN	O	O
during	NN	O	O
an	NN	O	O
in	NN	O	O
vivo	NN	O	O
chronic	NN	O	O
response	NN	O	O
,	NN	O	O
a	NN	O	O
narrow	NN	O	O
range	NN	O	O
of	NN	O	O
avidity	NN	O	O
of	NN	O	O
the	NN	O	O
TCR-CD8	NN	O	B-protein
complex	NN	O	I-protein
conditions	NN	O	O
long-term	NN	O	O
clonotype	NN	O	O
persistence	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
CD8	NN	O	B-protein
contribution	NN	O	O
is	NN	O	O
adjusted	NN	O	O
to	NN	O	O
keep	NN	O	O
clonotypes	NN	O	O
with	NN	O	O
variable	NN	O	O
TCR	NN	O	B-protein
affinities	NN	O	O
within	NN	O	O
this	NN	O	O
avidity	NN	O	O
window	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	O
localization	NN	O	O
and	NN	O	O
formation	NN	O	O
of	NN	O	O
beta-catenin-lymphoid	NN	O	B-protein
enhancer	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
complexes	NN	O	I-protein
are	NN	O	O
not	NN	O	O
sufficient	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
response	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
Wnt	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
,	NN	O	O
beta-catenin	NN	O	B-protein
accumulates	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
where	NN	O	O
it	NN	O	O
cooperates	NN	O	O
with	NN	O	O
LEF/TCF	NN	O	B-protein
(	NN	O	O
for	NN	O	O
lymphoid	NN	O	B-protein
enhancer	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	I-protein
T-cell	NN	O	I-protein
factor	NN	O	I-protein
)	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
activate	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
beta-catenin	NN	O	B-protein
undergoes	NN	O	O
this	NN	O	O
shift	NN	O	O
in	NN	O	O
location	NN	O	O
and	NN	O	O
participates	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
that	NN	O	O
beta-catenin	NN	O	B-protein
can	NN	O	O
be	NN	O	O
imported	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
independently	NN	O	O
of	NN	O	O
LEF/TCF	NN	O	B-protein
binding	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
may	NN	O	O
also	NN	O	O
be	NN	O	O
exported	NN	O	O
from	NN	O	O
nuclei	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
introduced	NN	O	O
a	NN	O	O
small	NN	O	O
deletion	NN	O	O
within	NN	O	O
beta-catenin	NN	O	B-protein
(	NN	O	O
Delta19	NN	O	B-DNA
)	NN	O	O
that	NN	O	O
disrupts	NN	O	O
binding	NN	O	O
to	NN	O	O
LEF-1	NN	O	B-protein
,	NN	O	O
E-cadherin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
APC	NN	O	B-protein
but	NN	O	O
not	NN	O	O
axin	NN	O	B-protein
.	NN	O	O

This	NN	O	O
Delta19	NN	O	B-protein
beta-catenin	NN	O	I-protein
mutant	NN	O	I-protein
localizes	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
because	NN	O	O
it	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
efficiently	NN	O	O
sequestered	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

The	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
Delta19	NN	O	B-DNA
definitively	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
beta-catenin	NN	O	B-protein
localizes	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
are	NN	O	O
completely	NN	O	O
independent	NN	O	O
of	NN	O	O
LEF/TCF	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

beta-Catenin	NN	O	B-protein
and	NN	O	O
LEF-1	NN	O	B-protein
complexes	NN	O	I-protein
can	NN	O	O
activate	NN	O	O
reporter	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
a	NN	O	O
transformed	NN	O	B-cell_line
T-lymphocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
Jurkat	NN	O	B-cell_line
)	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
even	NN	O	O
though	NN	O	O
both	NN	O	O
factors	NN	O	O
are	NN	O	O
nuclear	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
localization	NN	O	O
of	NN	O	O
both	NN	O	O
factors	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Excess	NN	O	O
beta-catenin	NN	O	B-protein
can	NN	O	O
squelch	NN	O	O
reporter	NN	O	O
gene	NN	O	O
activation	NN	O	O
by	NN	O	O
LEF-1-beta-catenin	NN	O	B-protein
complexes	NN	O	I-protein
but	NN	O	O
not	NN	O	O
activation	NN	O	O
by	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
VP16	NN	O	B-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
third	NN	O	O
component	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
gene	NN	O	O
activation	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
third	NN	O	O
component	NN	O	O
may	NN	O	O
vary	NN	O	O
with	NN	O	O
cell	NN	O	O
type	NN	O	O
.	NN	O	O

-DOCSTART-	O

Control	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
by	NN	O	O
the	NN	O	O
Ikaros	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
.	NN	O	O

Lymphoid	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
relies	NN	O	O
on	NN	O	O
precisely	NN	O	O
orchestrated	NN	O	O
gene	NN	O	O
activation	NN	O	O
and	NN	O	O
repression	NN	O	O
events	NN	O	O
.	NN	O	O

Gene	NN	O	O
targeting	NN	O	O
studies	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
crucial	NN	O	O
roles	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
Ikaros	NN	O	B-protein
and	NN	O	O
Aiolos	NN	O	B-protein
in	NN	O	O
regulating	NN	O	O
multiple	NN	O	O
stages	NN	O	O
of	NN	O	O
B	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

Recent	NN	O	O
experiments	NN	O	O
suggest	NN	O	O
that	NN	O	O
Ikaros	NN	O	B-protein
and	NN	O	O
Aiolos	NN	O	B-protein
set	NN	O	O
B	NN	O	O
cell	NN	O	O
antigen-receptor	NN	O	O
(	NN	O	O
BCR	NN	O	O
)	NN	O	O
-	NN	O	O
and	NN	O	O
TCR-mediated	NN	O	O
signaling	NN	O	O
thresholds	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
molecules	NN	O	O
exist	NN	O	O
within	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
nuclear	NN	O	B-protein
complexes	NN	O	I-protein
that	NN	O	O
contain	NN	O	O
nucleosome	NN	O	O
remodeling	NN	O	O
and	NN	O	O
histone	NN	O	B-protein
deacetylase	NN	O	I-protein
activities	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
targeting	NN	O	O
of	NN	O	O
retroviral	NN	O	O
vectors	NN	O	O
to	NN	O	O
the	NN	O	O
erythroblastic	NN	O	B-cell_type
progeny	NN	O	I-cell_type
of	NN	O	O
transduced	NN	O	B-cell_line
hematopoietic	NN	O	I-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Targeted	NN	O	O
expression	NN	O	O
to	NN	O	O
specific	NN	O	O
tissues	NN	O	O
or	NN	O	O
cell	NN	O	O
lineages	NN	O	O
is	NN	O	O
a	NN	O	O
necessary	NN	O	O
feature	NN	O	O
of	NN	O	O
a	NN	O	O
gene	NN	O	O
therapy	NN	O	O
vector	NN	O	O
for	NN	O	O
many	NN	O	O
clinical	NN	O	O
applications	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
correction	NN	O	O
of	NN	O	O
hemoglobinopathies	NN	O	O
or	NN	O	O
thalassemias	NN	O	O
by	NN	O	O
transplantation	NN	O	O
of	NN	O	O
genetically	NN	O	B-cell_type
modified	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
developed	NN	O	O
retroviral	NN	O	O
vectors	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
constitutive	NN	O	B-DNA
viral	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
U3	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
LTR	NN	O	I-DNA
is	NN	O	O
replaced	NN	O	O
by	NN	O	O
an	NN	O	O
autoregulatory	NN	O	B-DNA
enhancer	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
erythroid-specific	NN	O	B-DNA
GATA-1	NN	O	I-DNA
transcription	NN	O	I-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
replaced	NN	O	B-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
propagated	NN	O	O
to	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
LTR	NN	O	I-DNA
upon	NN	O	O
integration	NN	O	O
into	NN	O	O
the	NN	O	O
target	NN	O	B-DNA
cell	NN	O	I-DNA
genome	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
modified	NN	O	O
vectors	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
transduce	NN	O	O
human	NN	O	B-cell_line
hematopoietic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
cord	NN	O	B-cell_type
blood-derived	NN	O	I-cell_type
CD34	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
stem/progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
murine	NN	O	B-cell_line
bone	NN	O	I-cell_line
marrow	NN	O	I-cell_line
repopulating	NN	O	I-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
appropriate	NN	O	O
reporter	NN	O	B-DNA
genes	NN	O	I-DNA
(	NN	O	O
triangle	NN	O	B-DNA
upLNGFR	NN	O	I-DNA
,	NN	O	O
EGFP	NN	O	B-DNA
)	NN	O	O
was	NN	O	O
analyzed	NN	O	O
in	NN	O	O
the	NN	O	O
differentiated	NN	O	O
progeny	NN	O	O
of	NN	O	O
transduced	NN	O	B-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
in	NN	O	O
liquid	NN	O	O
culture	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
clonogenic	NN	O	O
assay	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
after	NN	O	O
bone	NN	O	O
marrow	NN	O	O
transplantation	NN	O	O
in	NN	O	O
lethally	NN	O	O
irradiated	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
GATA-1	NN	O	B-DNA
autoregulatory	NN	O	I-DNA
enhancer	NN	O	I-DNA
effectively	NN	O	O
restricts	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
LTR-driven	NN	O	B-DNA
proviral	NN	O	I-DNA
transcription	NN	O	I-DNA
unit	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
erythroblastic	NN	O	B-cell_type
progeny	NN	O	I-cell_type
of	NN	O	O
both	NN	O	O
human	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
and	NN	O	O
mouse-repopulating	NN	O	B-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Packaging	NN	O	O
of	NN	O	O
viral	NN	O	O
particles	NN	O	O
,	NN	O	O
integration	NN	O	O
into	NN	O	O
the	NN	O	O
target	NN	O	O
genome	NN	O	O
,	NN	O	O
and	NN	O	O
stability	NN	O	O
of	NN	O	O
the	NN	O	O
integrated	NN	O	O
provirus	NN	O	O
are	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
modification	NN	O	O
.	NN	O	O

Enhancer	NN	O	O
replacement	NN	O	O
is	NN	O	O
therefore	NN	O	O
an	NN	O	O
effective	NN	O	O
strategy	NN	O	O
to	NN	O	O
target	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
retroviral	NN	O	B-DNA
transgene	NN	O	I-DNA
to	NN	O	O
a	NN	O	O
specific	NN	O	O
progeny	NN	O	O
of	NN	O	O
transduced	NN	O	B-cell_line
hematopoietic	NN	O	I-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
cell	NN	O	O
death	NN	O	O
by	NN	O	O
apoptosis-linked	NN	O	B-DNA
gene	NN	O	I-DNA
4	NN	O	I-DNA
.	NN	O	O

Programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
is	NN	O	O
a	NN	O	O
process	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
normal	NN	O	O
development	NN	O	O
of	NN	O	O
an	NN	O	O
organism	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
best	NN	O	O
understood	NN	O	O
apoptotic	NN	O	O
pathways	NN	O	O
occurs	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
Fas	NN	O	B-protein
/Fas	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
FasL	NN	O	B-protein
)	NN	O	O
interaction	NN	O	O
.	NN	O	O

During	NN	O	O
studies	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
T	NN	O	O
cell-receptor	NN	O	O
engagement	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
ALG-4F	NN	O	B-protein
,	NN	O	O
a	NN	O	O
truncated	NN	O	B-protein
transcript	NN	O	I-protein
that	NN	O	O
prevents	NN	O	O
T	NN	O	O
cell-receptor-induced	NN	O	O
FasL	NN	O	B-protein
upregulation	NN	O	O
and	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
full-length	NN	O	B-protein
ALG-4	NN	O	I-protein
induced	NN	O	O
transcription	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
and	NN	O	O
,	NN	O	O
consequently	NN	O	O
,	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
ALG-4	NN	O	B-protein
is	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Fas/	NN	O	O
FasL	NN	O	B-protein
interaction	NN	O	O
initiates	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
many	NN	O	O
other	NN	O	O
systems	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
dysregulation	NN	O	O
is	NN	O	O
a	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
several	NN	O	O
pathologic	NN	O	O
conditions	NN	O	O
arise	NN	O	O
.	NN	O	O

Understanding	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
regulation	NN	O	O
could	NN	O	O
be	NN	O	O
very	NN	O	O
useful	NN	O	O
in	NN	O	O
elucidating	NN	O	O
how	NN	O	O
these	NN	O	O
diseases	NN	O	O
develop	NN	O	O
and	NN	O	O
in	NN	O	O
identifying	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
targets	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
oncoprotein	NN	O	I-protein
latent	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
engages	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-associated	NN	O	I-protein
proteins	NN	O	I-protein
TRADD	NN	O	B-protein
and	NN	O	O
receptor-interacting	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
RIP	NN	O	B-protein
)	NN	O	O
but	NN	O	O
does	NN	O	O
not	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
or	NN	O	O
require	NN	O	O
RIP	NN	O	B-protein
for	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

A	NN	O	O
site	NN	O	O
in	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
(	NN	O	I-protein
EBV	NN	O	I-protein
)	NN	O	I-protein
transforming	NN	O	I-protein
protein	NN	O	I-protein
LMP1	NN	O	B-protein
that	NN	O	O
constitutively	NN	O	O
associates	NN	O	O
with	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
TNFR1	NN	O	I-protein
)	NN	O	I-protein
-associated	NN	O	I-protein
death	NN	O	I-protein
domain	NN	O	I-protein
protein	NN	O	I-protein
TRADD	NN	O	B-protein
to	NN	O	O
mediate	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
activation	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
long-term	NN	O	O
lymphoblastoid	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
find	NN	O	O
that	NN	O	O
LMP1	NN	O	B-protein
signaling	NN	O	O
through	NN	O	O
TRADD	NN	O	B-protein
differs	NN	O	O
from	NN	O	O
TNFR1	NN	O	B-protein
signaling	NN	O	O
through	NN	O	O
TRADD	NN	O	B-protein
.	NN	O	O

LMP1	NN	O	B-protein
needs	NN	O	O
only	NN	O	O
11	NN	O	O
amino	NN	O	O
acids	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappaB	NN	O	B-protein
or	NN	O	O
synergize	NN	O	O
with	NN	O	O
TRADD	NN	O	B-protein
in	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
while	NN	O	O
TNFR1	NN	O	B-protein
requires	NN	O	O
approximately	NN	O	O
70	NN	O	O
residues	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
LMP1	NN	O	B-protein
does	NN	O	O
not	NN	O	O
require	NN	O	O
TRADD	NN	O	B-protein
residues	NN	O	O
294	NN	O	O
to	NN	O	O
312	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
while	NN	O	O
TNFR1	NN	O	B-protein
requires	NN	O	O
TRADD	NN	O	B-protein
residues	NN	O	O
296	NN	O	O
to	NN	O	O
302	NN	O	O
.	NN	O	O

LMP1	NN	O	B-protein
is	NN	O	O
partially	NN	O	O
blocked	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
a	NN	O	O
TRADD	NN	O	B-protein
mutant	NN	O	O
consisting	NN	O	O
of	NN	O	O
residues	NN	O	B-protein
122	NN	O	I-protein
to	NN	O	I-protein
293	NN	O	I-protein
.	NN	O	O

Unlike	NN	O	O
TNFR1	NN	O	B-protein
,	NN	O	O
LMP1	NN	O	B-protein
can	NN	O	O
interact	NN	O	O
directly	NN	O	O
with	NN	O	O
receptor-interacting	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
RIP	NN	O	B-protein
)	NN	O	O
and	NN	O	O
stably	NN	O	O
associates	NN	O	O
with	NN	O	O
RIP	NN	O	B-protein
in	NN	O	O
EBV-transformed	NN	O	B-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
LMP1	NN	O	B-protein
does	NN	O	O
not	NN	O	O
require	NN	O	O
RIP	NN	O	B-protein
for	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Despite	NN	O	O
constitutive	NN	O	O
association	NN	O	O
with	NN	O	O
TRADD	NN	O	B-protein
or	NN	O	O
RIP	NN	O	B-protein
,	NN	O	O
LMP1	NN	O	B-protein
does	NN	O	O
not	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
EBV-negative	NN	O	B-cell_line
Burkitt	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
or	NN	O	I-cell_line
human	NN	O	I-cell_line
embryonic	NN	O	I-cell_line
kidney	NN	O	I-cell_line
293	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
add	NN	O	O
a	NN	O	O
different	NN	O	O
perspective	NN	O	O
to	NN	O	O
the	NN	O	O
molecular	NN	O	O
interactions	NN	O	O
through	NN	O	O
which	NN	O	O
LMP1	NN	O	B-protein
,	NN	O	O
TRADD	NN	O	B-protein
,	NN	O	O
and	NN	O	O
RIP	NN	O	B-protein
participate	NN	O	O
in	NN	O	O
B-lymphocyte	NN	O	O
activation	NN	O	O
and	NN	O	O
growth	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
human	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
induces	NN	O	O
caspase-dependent	NN	O	O
apoptosis	NN	O	O
without	NN	O	O
detectable	NN	O	O
activation	NN	O	O
of	NN	O	O
caspase-1	NN	O	B-protein
and	NN	O	I-protein
-3	NN	O	I-protein
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
various	NN	O	B-DNA
genes	NN	O	I-DNA
that	NN	O	O
act	NN	O	O
as	NN	O	O
extrinsic	NN	O	O
and	NN	O	O
intrinsic	NN	O	O
survival	NN	O	O
factors	NN	O	O
for	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
show	NN	O	O
that	NN	O	O
suppression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
with	NN	O	O
cell-permeable	NN	O	O
SN50	NN	O	O
peptide	NN	O	O
,	NN	O	O
which	NN	O	O
masks	NN	O	O
the	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
sequence	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
dimers	NN	O	I-protein
and	NN	O	O
prevents	NN	O	O
their	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
,	NN	O	O
induces	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
normal	NN	O	I-cell_type
human	NN	O	I-cell_type
PBL	NN	O	I-cell_type
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
externalization	NN	O	O
of	NN	O	O
phosphatidylserine	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
DNA	NN	O	O
breaks	NN	O	O
,	NN	O	O
and	NN	O	O
morphological	NN	O	O
changes	NN	O	O
consistent	NN	O	O
with	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

DNA	NN	O	O
fragmentation	NN	O	O
was	NN	O	O
efficiently	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
caspase	NN	O	B-protein
inhibitor	NN	O	I-protein
Z-VAD-fmk	NN	O	B-protein
and	NN	O	O
partially	NN	O	O
blocked	NN	O	O
by	NN	O	O
Ac-DEVD-fmk	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
SN50-mediated	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
caspase	NN	O	B-protein
-dependent	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
suppression	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
that	NN	O	O
induced	NN	O	O
by	NN	O	O
TPEN	NN	O	O
(	NN	O	O
N	NN	O	O
,	NN	O	O
N	NN	O	O
,	NN	O	O
N	NN	O	O
'	NN	O	O
,	NN	O	O
N'-tetrakis	NN	O	O
[	NN	O	O
2-pyridylmethyl	NN	O	O
]	NN	O	O
ethylenediamine	NN	O	O
)	NN	O	O
or	NN	O	O
soluble	NN	O	B-protein
Fas	NN	O	I-protein
ligand	NN	O	I-protein
(	NN	O	O
CD95	NN	O	B-protein
)	NN	O	O
,	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
active	NN	O	B-protein
death	NN	O	I-protein
effector	NN	O	I-protein
proteases	NN	O	I-protein
caspase-1	NN	O	B-protein
-like	NN	O	O
(	NN	O	O
IL-1	NN	O	B-protein
converting	NN	O	I-protein
enzyme	NN	O	I-protein
)	NN	O	O
,	NN	O	O
caspase-3	NN	O	B-protein
-like	NN	O	O
(	NN	O	O
CPP32/Yama/apopain	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
caspase-6	NN	O	B-protein
-like	NN	O	O
and	NN	O	O
without	NN	O	O
cleavage	NN	O	O
of	NN	O	O
caspase-3	NN	O	B-protein
substrates	NN	O	I-protein
poly	NN	O	B-protein
(	NN	O	I-protein
ADP-ribose	NN	O	I-protein
)	NN	O	I-protein
polymerase	NN	O	I-protein
and	NN	O	O
DNA	NN	O	B-protein
fragmentation	NN	O	I-protein
factor-45	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
either	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
activation	NN	O	O
is	NN	O	O
required	NN	O	O
or	NN	O	O
that	NN	O	O
different	NN	O	O
caspases	NN	O	B-protein
are	NN	O	O
involved	NN	O	O
.	NN	O	O

Preactivation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
resulting	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
nuclear	NN	O	O
translocation	NN	O	O
protected	NN	O	O
cells	NN	O	O
from	NN	O	O
SN50-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
survival	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
PBL	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
mediators	NN	O	O
:	NN	O	O
comparative	NN	O	O
study	NN	O	O
with	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
neutrophils	NN	O	I-cell_type
or	NN	O	O
transformed	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
putative	NN	O	O
roles	NN	O	O
in	NN	O	O
functionality	NN	O	O
and	NN	O	O
cell	NN	O	O
biology	NN	O	O
.	NN	O	O

Granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-3	NN	O	I-protein
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
92-kDa	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
identified	NN	O	O
this	NN	O	O
92-kDa	NN	O	B-protein
protein	NN	O	I-protein
as	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
as	NN	O	O
STATs1	NN	O	B-protein
,	NN	O	I-protein
3	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
6	NN	O	I-protein
nor	NN	O	O
c-fes	NN	O	B-protein
and	NN	O	O
vav	NN	O	B-protein
protooncogene	NN	O	I-protein
products	NN	O	I-protein
,	NN	O	O
and	NN	O	O
demonstrated	NN	O	O
its	NN	O	O
translocation	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
enhancement	NN	O	O
of	NN	O	O
specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
capacity	NN	O	O
,	NN	O	O
and	NN	O	O
potentiation	NN	O	O
of	NN	O	O
trancriptional	NN	O	O
activity	NN	O	O
by	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

N-formyl-methionyl-leucyl-phenylalanine	NN	O	O
(	NN	O	O
FMLP	NN	O	O
)	NN	O	O
and	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
42-	NN	O	B-protein
and	NN	O	I-protein
44-kDa	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
which	NN	O	O
were	NN	O	O
identified	NN	O	O
as	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
ERK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
marked	NN	O	O
contrast	NN	O	O
to	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
MO7e	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
GM-CSF	NN	O	B-protein
did	NN	O	O
not	NN	O	O
induce	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
ERK	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Among	NN	O	O
upstream	NN	O	B-protein
signaling	NN	O	I-protein
molecules	NN	O	I-protein
of	NN	O	O
ERK	NN	O	B-protein
,	NN	O	O
Shc	NN	O	B-protein
was	NN	O	O
constitutively	NN	O	O
associated	NN	O	O
with	NN	O	O
Grb2	NN	O	O
and	NN	O	O
was	NN	O	O
not	NN	O	O
tyrosine-phosphorylated	NN	O	O
by	NN	O	O
GM-CSF	NN	O	B-protein
and	NN	O	O
FMLP	NN	O	O
,	NN	O	O
and	NN	O	O
Sos1	NN	O	B-protein
and	NN	O	O
c-Raf-1	NN	O	B-protein
were	NN	O	O
not	NN	O	O
phosphorylated	NN	O	O
by	NN	O	O
GM-CSF	NN	O	B-protein
,	NN	O	O
IL-3	NN	O	O
,	NN	O	O
TNF	NN	O	O
,	NN	O	O
and	NN	O	O
FMLP	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
whereas	NN	O	O
all	NN	O	O
these	NN	O	O
signaling	NN	O	O
molecules	NN	O	O
were	NN	O	O
affected	NN	O	O
and/or	NN	O	O
utilized	NN	O	O
by	NN	O	O
GM-CSF	NN	O	B-protein
in	NN	O	O
MO7e	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
p38	NN	O	B-protein
was	NN	O	O
constitutively	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
agonist-dependent	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
was	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Superoxide	NN	O	O
release	NN	O	O
stimulated	NN	O	O
by	NN	O	O
FMLP	NN	O	O
was	NN	O	O
inhibited	NN	O	O
partially	NN	O	O
by	NN	O	O
PD98059	NN	O	O
or	NN	O	O
SB203580	NN	O	O
,	NN	O	O
a	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
ERK	NN	O	O
or	NN	O	O
p38	NN	O	O
pathway	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
almost	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
both	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
whereas	NN	O	O
PMA-induced	NN	O	O
superoxide	NN	O	O
release	NN	O	O
was	NN	O	O
resistant	NN	O	O
to	NN	O	O
these	NN	O	O
two	NN	O	O
inhibitors	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

PD98059	NN	O	O
inhibited	NN	O	O
GM-CSF	NN	O	B-protein
-dependent	NN	O	O
proliferation	NN	O	O
of	NN	O	O
MO7e	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Present	NN	O	O
results	NN	O	O
indicate	NN	O	O
trancriptional	NN	O	O
roles	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
and	NN	O	O
functional	NN	O	O
roles	NN	O	O
of	NN	O	O
ERK	NN	O	B-protein
and/or	NN	O	O
p38	NN	O	B-protein
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
physiological	NN	O	O
receptor-mediated	NN	O	O
agonists	NN	O	O
GM-CSF	NN	O	B-protein
and	NN	O	O
FMLP	NN	O	O
.	NN	O	O

Possible	NN	O	O
roles	NN	O	O
of	NN	O	O
ERK	NN	O	B-protein
in	NN	O	O
proliferation	NN	O	O
of	NN	O	O
transformed	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
also	NN	O	O
suggested	NN	O	O
.	NN	O	O

-DOCSTART-	O

Aiolos	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
controls	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
regulating	NN	O	O
Bcl-2	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
its	NN	O	O
cellular	NN	O	O
localization	NN	O	O
.	NN	O	O

We	NN	O	O
searched	NN	O	O
for	NN	O	O
proteins	NN	O	O
that	NN	O	O
interact	NN	O	O
with	NN	O	O
Ras	NN	O	B-protein
in	NN	O	O
interleukin	NN	O	B-cell_line
(	NN	O	I-cell_line
IL	NN	O	I-cell_line
)	NN	O	I-cell_line
-2-stimulated	NN	O	I-cell_line
or	NN	O	I-cell_line
IL-2-deprived	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Aiolos	NN	O	B-protein
interacts	NN	O	O
with	NN	O	O
Ras	NN	O	B-protein
.	NN	O	O

The	NN	O	O
Ras	NN	O	B-protein
-Aiolos	NN	O	B-protein
interaction	NN	O	O
was	NN	O	O
confirmed	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
by	NN	O	O
co-immunoprecipitation	NN	O	O
.	NN	O	O

Indirect	NN	O	O
immunofluorescence	NN	O	O
shows	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
controls	NN	O	O
the	NN	O	O
cellular	NN	O	O
distribution	NN	O	O
of	NN	O	O
Aiolos	NN	O	B-protein
and	NN	O	O
induces	NN	O	O
its	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
required	NN	O	O
for	NN	O	O
dissociation	NN	O	O
from	NN	O	O
Ras	NN	O	B-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
identified	NN	O	O
functional	NN	O	O
Aiolos-binding	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
Bcl-2	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
are	NN	O	O
able	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
Aiolos-binding	NN	O	B-DNA
sites	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
Bcl-2	NN	O	B-DNA
promoter	NN	O	I-DNA
inhibits	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
luciferase	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
direct	NN	O	O
control	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
Aiolos	NN	O	B-protein
.	NN	O	O

Co-transfection	NN	O	O
experiments	NN	O	O
confirm	NN	O	O
that	NN	O	O
Aiolos	NN	O	B-protein
induces	NN	O	O
Bcl-2	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
prevents	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
IL-2-deprived	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
propose	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
expression	NN	O	O
via	NN	O	O
Aiolos	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Binding	NN	O	O
of	NN	O	O
YY1	NN	O	B-protein
and	NN	O	O
Oct1	NN	O	B-protein
to	NN	O	O
a	NN	O	O
novel	NN	O	O
element	NN	O	O
that	NN	O	O
downregulates	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
in	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
IL-5	NN	O	B-protein
controls	NN	O	O
development	NN	O	O
of	NN	O	O
eosinophilia	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
allergic	NN	O	O
diseases	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
atopic	NN	O	O
and	NN	O	O
nonatopic	NN	O	O
asthma	NN	O	O
,	NN	O	O
elevated	NN	O	O
IL-5	NN	O	B-protein
has	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
and	NN	O	O
the	NN	O	O
airways	NN	O	O
.	NN	O	O

IL-5	NN	O	B-protein
is	NN	O	O
produced	NN	O	O
mainly	NN	O	O
by	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
its	NN	O	O
expression	NN	O	O
is	NN	O	O
regulated	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
This	NN	O	O
study	NN	O	O
focuses	NN	O	O
on	NN	O	O
the	NN	O	O
functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-5	NN	O	I-DNA
(	NN	O	I-DNA
hIL-5	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
cis	NN	O	O
-regulatory	NN	O	O
elements	NN	O	O
and	NN	O	O
transcription	NN	O	O
factors	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Methods	NN	O	O
used	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
include	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
footprint	NN	O	O
assays	NN	O	O
,	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
and	NN	O	O
functional	NN	O	O
analysis	NN	O	O
by	NN	O	O
mammalian	NN	O	O
cell	NN	O	O
transfection	NN	O	O
involving	NN	O	O
deletion	NN	O	O
analysis	NN	O	O
and	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
We	NN	O	O
identified	NN	O	O
5	NN	O	O
protein	NN	O	B-DNA
binding	NN	O	I-DNA
regions	NN	O	I-DNA
(	NN	O	O
BRs	NN	O	B-DNA
)	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
hIL-5	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Functional	NN	O	O
analysis	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
BRs	NN	O	B-DNA
are	NN	O	O
involved	NN	O	O
in	NN	O	O
control	NN	O	O
of	NN	O	O
hIL-5	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

Two	NN	O	O
of	NN	O	O
these	NN	O	O
regions	NN	O	O
,	NN	O	O
BR3	NN	O	B-DNA
and	NN	O	O
BR4	NN	O	B-DNA
located	NN	O	O
at	NN	O	O
positions	NN	O	O
-102	NN	O	O
to	NN	O	O
-73	NN	O	O
,	NN	O	O
have	NN	O	O
not	NN	O	O
previously	NN	O	O
been	NN	O	O
described	NN	O	O
as	NN	O	O
regulators	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
BR3	NN	O	B-DNA
sequence	NN	O	O
contains	NN	O	O
a	NN	O	O
novel	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
located	NN	O	O
at	NN	O	O
positions	NN	O	O
-90	NN	O	O
to	NN	O	O
-79	NN	O	O
of	NN	O	O
the	NN	O	O
hIL-5	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
binds	NN	O	O
Oct1	NN	O	B-protein
,	NN	O	O
octamer-like	NN	O	B-protein
,	NN	O	O
and	NN	O	O
YY1	NN	O	B-protein
nuclear	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Substitution	NN	O	O
mutations	NN	O	O
,	NN	O	O
which	NN	O	O
abolished	NN	O	O
binding	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
to	NN	O	O
the	NN	O	O
BR3	NN	O	B-DNA
sequence	NN	O	I-DNA
,	NN	O	O
significantly	NN	O	O
increased	NN	O	O
hIL-5	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
We	NN	O	O
suggest	NN	O	O
that	NN	O	O
Oct1	NN	O	B-protein
,	NN	O	O
YY1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
octamer-like	NN	O	B-protein
factors	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
-90/-79	NN	O	B-DNA
sequence	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
IL-5	NN	O	I-DNA
promoter	NN	O	I-DNA
are	NN	O	O
involved	NN	O	O
in	NN	O	O
suppression	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

New	NN	O	O
immunosuppressive	NN	O	O
drug	NN	O	O
PNU156804	NN	O	O
blocks	NN	O	O
IL-2	NN	O	B-protein
-dependent	NN	O	O
proliferation	NN	O	O
and	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
and	NN	O	O
AP-1	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
had	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
drug	NN	O	O
undecylprodigiosin	NN	O	O
(	NN	O	O
UP	NN	O	O
)	NN	O	O
blocks	NN	O	O
human	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
proliferation	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
investigated	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
analogue	NN	O	O
of	NN	O	O
UP	NN	O	O
,	NN	O	O
PNU156804	NN	O	O
,	NN	O	O
which	NN	O	O
shows	NN	O	O
a	NN	O	O
more	NN	O	O
favorable	NN	O	O
activity	NN	O	O
profile	NN	O	O
than	NN	O	O
UP	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
biological	NN	O	O
effect	NN	O	O
of	NN	O	O
PNU156804	NN	O	O
in	NN	O	O
vitro	NN	O	O
is	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
UP	NN	O	O
:	NN	O	O
PNU156804	NN	O	O
blocks	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
in	NN	O	O
mid-late	NN	O	O
G1	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
cell	NN	O	O
cycle	NN	O	O
analysis	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
cyclins	NN	O	B-protein
,	NN	O	O
and	NN	O	O
cyclin-dependent	NN	O	B-protein
kinases	NN	O	I-protein
and	NN	O	O
retinoblastoma	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
PNU156804	NN	O	O
does	NN	O	O
not	NN	O	O
block	NN	O	O
significantly	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
either	NN	O	O
IL-2	NN	O	B-protein
or	NN	O	O
IL-2R	NN	O	B-protein
alpha-	NN	O	I-protein
and	NN	O	I-protein
gamma-chains	NN	O	I-protein
but	NN	O	O
inhibits	NN	O	O
IL-2	NN	O	B-protein
-dependent	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
several	NN	O	O
molecular	NN	O	O
pathways	NN	O	O
that	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
PNU156804	NN	O	O
does	NN	O	O
not	NN	O	O
inhibit	NN	O	O
c-myc	NN	O	O
and	NN	O	O
bcl-2	NN	O	O
mRNA	NN	O	O
induction	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
PNU156804	NN	O	O
efficiently	NN	O	O
inhibits	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	O
factors	NN	O	O
.	NN	O	O

PNU156804	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-beta	NN	O	I-protein
.	NN	O	O

PNU156804	NN	O	O
action	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
some	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
;	NN	O	O
it	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
PMA	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
blocks	NN	O	O
that	NN	O	O
induced	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
cross-linking	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
prodigiosin	NN	O	O
family	NN	O	O
of	NN	O	O
immunosuppressants	NN	O	O
is	NN	O	O
a	NN	O	O
new	NN	O	O
family	NN	O	O
of	NN	O	O
molecules	NN	O	O
that	NN	O	O
show	NN	O	O
a	NN	O	O
novel	NN	O	O
target	NN	O	O
specificity	NN	O	O
clearly	NN	O	O
distinct	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
other	NN	O	O
immunosuppressive	NN	O	O
drugs	NN	O	O
such	NN	O	O
as	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
FK506	NN	O	O
,	NN	O	O
and	NN	O	O
rapamycin	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
Tax	NN	O	I-protein
protein	NN	O	I-protein
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
STAT1	NN	O	B-DNA
and	NN	O	I-DNA
STAT5	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
T-cells	NN	O	B-cell_type
.	NN	O	O

Human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
HTLV-1	NN	O	I-protein
)	NN	O	I-protein
Tax	NN	O	I-protein
transforms	NN	O	O
normal	NN	O	O
T-cells	NN	O	B-cell_type
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

STAT	NN	O	B-protein
is	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
play	NN	O	O
a	NN	O	O
pivotal	NN	O	O
role	NN	O	O
in	NN	O	O
cytokine-induced	NN	O	O
functions	NN	O	O
of	NN	O	O
a	NN	O	O
various	NN	O	O
type	NN	O	O
of	NN	O	O
cells	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
STATs	NN	O	B-protein
in	NN	O	O
the	NN	O	O
transformation	NN	O	O
of	NN	O	O
T-cells	NN	O	B-cell_type
by	NN	O	O
HTLV-1	NN	O	O
.	NN	O	O

HTLV-1-transformed	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
expressed	NN	O	O
higher	NN	O	O
amounts	NN	O	O
of	NN	O	O
STAT1	NN	O	B-RNA
,	NN	O	I-RNA
STAT3	NN	O	I-RNA
and	NN	O	I-RNA
STAT5	NN	O	I-RNA
RNA	NN	O	I-RNA
and	NN	O	O
proteins	NN	O	O
than	NN	O	O
virus-negative	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
was	NN	O	O
induced	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

IL-2	NN	O	B-protein
induced	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
of	NN	O	O
a	NN	O	O
HTLV-1-transformed	NN	O	O
cell	NN	O	O
line	NN	O	O
and	NN	O	O
then	NN	O	O
stimulated	NN	O	O
its	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
did	NN	O	O
neither	NN	O	O
in	NN	O	O
a	NN	O	O
cell	NN	O	O
line	NN	O	O
lacking	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
STAT1	NN	O	B-RNA
,	NN	O	I-RNA
STAT3	NN	O	I-RNA
and	NN	O	I-RNA
STAT5	NN	O	I-RNA
mRNAs	NN	O	I-RNA
were	NN	O	O
also	NN	O	O
induced	NN	O	O
by	NN	O	O
a	NN	O	O
T-cell	NN	O	B-protein
mitogen	NN	O	I-protein
in	NN	O	O
normal	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
by	NN	O	O
Tax	NN	O	B-protein
enhances	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
functions	NN	O	O
of	NN	O	O
virus-infected	NN	O	O
T-cells	NN	O	B-cell_type
,	NN	O	O
hence	NN	O	O
the	NN	O	O
induction	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
-dependent	NN	O	O
transformation	NN	O	O
steps	NN	O	O
of	NN	O	O
T-cells	NN	O	B-cell_type
by	NN	O	O
HTLV-1	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
IL-5	NN	O	B-protein
gene	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
helper	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
IL-5	NN	O	B-protein
synthesis	NN	O	O
is	NN	O	O
regulated	NN	O	O
independently	NN	O	O
from	NN	O	O
IL-2	NN	O	B-protein
or	NN	O	O
IL-4	NN	O	B-protein
synthesis	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
IL-5	NN	O	B-protein
is	NN	O	O
fundamentally	NN	O	O
involved	NN	O	O
in	NN	O	O
eosinophilic	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Control	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
production	NN	O	O
may	NN	O	O
be	NN	O	O
effective	NN	O	O
for	NN	O	O
the	NN	O	O
management	NN	O	O
of	NN	O	O
allergic	NN	O	O
diseases	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
We	NN	O	O
aimed	NN	O	O
to	NN	O	O
find	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
regulate	NN	O	O
the	NN	O	O
IL-5	NN	O	B-protein
gene	NN	O	O
to	NN	O	O
selectively	NN	O	O
control	NN	O	O
IL-5	NN	O	B-protein
synthesis	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Allergen-specific	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
and	NN	O	O
T-cell	NN	O	B-cell_line
hybridomas	NN	O	I-cell_line
were	NN	O	O
established	NN	O	O
from	NN	O	O
the	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
asthma	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
investigated	NN	O	O
with	NN	O	O
transient	NN	O	O
transfection	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analysis	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
A	NN	O	O
human	NN	O	O
IL-5	NN	O	B-DNA
promoter/enhancer-luciferase	NN	O	I-DNA
gene	NN	O	I-DNA
construct	NN	O	I-DNA
,	NN	O	O
pIL-5	NN	O	B-DNA
(	NN	O	I-DNA
-511	NN	O	I-DNA
)	NN	O	I-DNA
Luc	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
transcribed	NN	O	O
on	NN	O	O
activation	NN	O	O
of	NN	O	O
IL-5-producing	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-5-nonproducing	NN	O	B-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

pIL-5	NN	O	B-DNA
(	NN	O	I-DNA
-511	NN	O	I-DNA
)	NN	O	I-DNA
Luc	NN	O	I-DNA
was	NN	O	O
transcribed	NN	O	O
by	NN	O	O
T-cell	NN	O	B-cell_line
hybridomas	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
fusion	NN	O	O
between	NN	O	O
IL-5-producing	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
and	NN	O	O
an	NN	O	O
IL-5	NN	O	B-cell_line
gene-nonexpressing	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
hybridomas	NN	O	B-cell_line
derived	NN	O	O
from	NN	O	O
IL-5-nonproducing	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

IL-5	NN	O	B-protein
synthesis	NN	O	O
was	NN	O	O
not	NN	O	O
only	NN	O	O
induced	NN	O	O
by	NN	O	O
T-cell	NN	O	O
receptor	NN	O	O
stimulation	NN	O	O
but	NN	O	O
also	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
was	NN	O	O
induced	NN	O	O
by	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
TcR	NN	O	B-protein
)	NN	O	O
stimulation	NN	O	O
,	NN	O	O
although	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
significant	NN	O	O
upregulation	NN	O	O
of	NN	O	O
binding	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
IL-5	NN	O	B-protein
synthesis	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
helper	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
regulated	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

A	NN	O	O
unique	NN	O	O
transcriptional	NN	O	O
mechanism	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
regulating	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
or	NN	O	I-DNA
IL-4	NN	O	I-DNA
genes	NN	O	I-DNA
seems	NN	O	O
to	NN	O	O
control	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Selective	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
may	NN	O	O
be	NN	O	O
useful	NN	O	O
for	NN	O	O
treating	NN	O	O
eosinophlic	NN	O	O
inflammation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Defining	NN	O	O
therapeutic	NN	O	O
targets	NN	O	O
by	NN	O	O
using	NN	O	O
adenovirus	NN	O	O
:	NN	O	O
blocking	NN	O	O
NF-kappaB	NN	O	B-protein
inhibits	NN	O	O
both	NN	O	O
inflammatory	NN	O	O
and	NN	O	O
destructive	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
rheumatoid	NN	O	O
synovium	NN	O	O
but	NN	O	O
spares	NN	O	O
anti-inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
has	NN	O	O
long	NN	O	O
been	NN	O	O
a	NN	O	O
subject	NN	O	O
of	NN	O	O
controversy	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
an	NN	O	O
adenoviral	NN	O	O
technique	NN	O	O
of	NN	O	O
blocking	NN	O	O
NF-kappaB	NN	O	B-protein
through	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	B-protein
subunit	NN	O	I-protein
IkappaBalpha	NN	O	B-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
the	NN	O	O
advantage	NN	O	O
that	NN	O	O
it	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
in	NN	O	O
the	NN	O	O
diseased	NN	O	O
tissue	NN	O	O
itself	NN	O	O
,	NN	O	O
with	NN	O	O
>	NN	O	O
90	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
synovial	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
fibroblasts	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
infected	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
spontaneous	NN	O	O
production	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
other	NN	O	O
pro-inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
is	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
in	NN	O	O
rheumatoid	NN	O	O
synovial	NN	O	O
tissue	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
main	NN	O	O
anti-inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
,	NN	O	O
like	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	I-protein
-11	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	I-protein
antagonist	NN	O	I-protein
.	NN	O	O

Of	NN	O	O
even	NN	O	O
more	NN	O	O
interest	NN	O	O
,	NN	O	O
IkappaBalpha	NN	O	B-protein
overexpression	NN	O	O
inhibited	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
matrix	NN	O	B-protein
metalloproteinases	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	I-protein
3	NN	O	I-protein
while	NN	O	O
not	NN	O	O
affecting	NN	O	O
their	NN	O	O
tissue	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

Blocking	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
rheumatoid	NN	O	O
joint	NN	O	O
thus	NN	O	O
has	NN	O	O
a	NN	O	O
very	NN	O	O
beneficial	NN	O	O
profile	NN	O	O
,	NN	O	O
reducing	NN	O	O
both	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
and	NN	O	O
the	NN	O	O
tissue	NN	O	O
destruction	NN	O	O
.	NN	O	O

The	NN	O	O
adenoviral	NN	O	O
technique	NN	O	O
described	NN	O	O
here	NN	O	O
has	NN	O	O
widespread	NN	O	O
applicability	NN	O	O
,	NN	O	O
allowing	NN	O	O
rapid	NN	O	O
testing	NN	O	O
of	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
blocking	NN	O	O
a	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
target	NN	O	O
in	NN	O	O
either	NN	O	O
cultures	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
in	NN	O	O
the	NN	O	O
diseased	NN	O	O
tissue	NN	O	O
itself	NN	O	O
.	NN	O	O

-DOCSTART-	O

Resistance	NN	O	O
to	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
vitro	NN	O	O
correlates	NN	O	O
with	NN	O	O
high	NN	O	O
metastatic	NN	O	O
capacity	NN	O	O
of	NN	O	O
cells	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

TNF	NN	O	B-protein
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
secreted	NN	O	O
by	NN	O	O
the	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
those	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
lacking	NN	O	O
capability	NN	O	O
to	NN	O	O
form	NN	O	O
metastatic	NN	O	O
tumors	NN	O	O
in	NN	O	O
vivo	NN	O	O
are	NN	O	O
susceptible	NN	O	O
to	NN	O	O
TNF	NN	O	B-protein
induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
with	NN	O	O
high	NN	O	O
metastatic	NN	O	O
potential	NN	O	O
are	NN	O	O
resistant	NN	O	O
to	NN	O	O
TNF	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
same	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
resistant	NN	O	O
to	NN	O	O
cytolytic	NN	O	O
action	NN	O	O
of	NN	O	O
other	NN	O	O
cytotoxic	NN	O	B-protein
proteins	NN	O	I-protein
secreted	NN	O	O
by	NN	O	O
LAK	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
data	NN	O	O
showed	NN	O	O
that	NN	O	O
TNF	NN	O	B-protein
resistance	NN	O	O
in	NN	O	O
vitro	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
increased	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

This	NN	O	O
finding	NN	O	O
may	NN	O	O
provide	NN	O	O
a	NN	O	O
tool	NN	O	O
to	NN	O	O
improve	NN	O	O
current	NN	O	O
protocols	NN	O	O
of	NN	O	O
immunotherapy	NN	O	O
and	NN	O	O
insights	NN	O	O
to	NN	O	O
how	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
or	NN	O	O
are	NN	O	O
not	NN	O	O
killed	NN	O	O
by	NN	O	O
LAK	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Clonality	NN	O	O
analysis	NN	O	O
of	NN	O	O
granulocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
healthy	NN	O	O
females	NN	O	O
by	NN	O	O
the	NN	O	O
PCR-based	NN	O	O
HUMARA	NN	O	B-DNA
method	NN	O	O
.	NN	O	O

Clonality	NN	O	O
analysis	NN	O	O
utilizing	NN	O	O
X-chromosome	NN	O	B-DNA
inactivation	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
in	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
various	NN	O	O
diseases	NN	O	O
,	NN	O	O
including	NN	O	O
hematological	NN	O	O
malignancies	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
androgen	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
HUMARA	NN	O	B-DNA
)	NN	O	O
assay	NN	O	O
is	NN	O	O
the	NN	O	O
newest	NN	O	O
of	NN	O	O
such	NN	O	O
methods	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
female	NN	O	O
population	NN	O	O
can	NN	O	O
be	NN	O	O
assessed	NN	O	O
by	NN	O	O
this	NN	O	O
relatively	NN	O	O
simple	NN	O	O
procedure	NN	O	O
.	NN	O	O

One	NN	O	O
problem	NN	O	O
in	NN	O	O
using	NN	O	O
these	NN	O	O
clonality	NN	O	O
analysis	NN	O	O
methods	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
is	NN	O	O
that	NN	O	O
there	NN	O	O
may	NN	O	O
be	NN	O	O
significant	NN	O	O
variation	NN	O	O
in	NN	O	O
Lyonization	NN	O	O
in	NN	O	O
blood	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
normal	NN	O	O
individuals	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
diversity	NN	O	O
in	NN	O	O
X-chromosome	NN	O	B-DNA
methylation	NN	O	O
patterns	NN	O	O
,	NN	O	O
which	NN	O	O
reflect	NN	O	O
Lyonization	NN	O	O
,	NN	O	O
assessed	NN	O	O
by	NN	O	O
the	NN	O	O
HUMARA	NN	O	B-DNA
assay	NN	O	O
in	NN	O	O
the	NN	O	O
supposedly	NN	O	O
normal	NN	O	O
population	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
granulocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
97	NN	O	O
relatively	NN	O	O
young	NN	O	O
(	NN	O	O
18-	NN	O	O
to	NN	O	O
35-year-old	NN	O	O
)	NN	O	O
healthy	NN	O	O
female	NN	O	O
volunteers	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
methylation	NN	O	O
patterns	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
HUMARA	NN	O	B-DNA
alleles	NN	O	I-DNA
were	NN	O	O
distributed	NN	O	O
even	NN	O	O
more	NN	O	O
widely	NN	O	O
,	NN	O	O
both	NN	O	O
in	NN	O	O
granuloctyes	NN	O	B-cell_type
and	NN	O	O
in	NN	O	O
T	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
than	NN	O	O
previously	NN	O	O
reported	NN	O	O
with	NN	O	O
other	NN	O	O
methods	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
deviation	NN	O	O
of	NN	O	O
methylation	NN	O	O
in	NN	O	O
granulocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
well	NN	O	O
correlated	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
appropriate	NN	O	O
controls	NN	O	O
from	NN	O	O
the	NN	O	O
same	NN	O	O
individuals	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
T	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
stem	NN	O	O
cell	NN	O	O
disorders	NN	O	O
,	NN	O	O
should	NN	O	O
always	NN	O	O
be	NN	O	O
employed	NN	O	O
to	NN	O	O
conclusively	NN	O	O
determine	NN	O	O
whether	NN	O	O
certain	NN	O	O
cells	NN	O	O
of	NN	O	O
hematopoietic	NN	O	O
origin	NN	O	O
are	NN	O	O
clonal	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
B-cell	NN	O	O
response	NN	O	O
in	NN	O	O
intrahepatic	NN	O	O
lymphoid	NN	O	O
follicles	NN	O	O
in	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
C	NN	O	O
.	NN	O	O

Intrahepatic	NN	O	O
lymphoid	NN	O	O
follicle	NN	O	O
(	NN	O	O
ILF	NN	O	O
)	NN	O	O
formation	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
characteristic	NN	O	O
and	NN	O	O
commonly	NN	O	O
observed	NN	O	O
histological	NN	O	O
features	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
C	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
regarding	NN	O	O
whether	NN	O	O
follicles	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
belong	NN	O	O
to	NN	O	O
functional	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
,	NN	O	O
where	NN	O	O
B	NN	O	O
cells	NN	O	O
are	NN	O	O
activated	NN	O	O
,	NN	O	O
differentiated	NN	O	O
,	NN	O	O
and	NN	O	O
proliferated	NN	O	O
,	NN	O	O
or	NN	O	O
if	NN	O	O
the	NN	O	O
lymphocytes	NN	O	O
are	NN	O	O
merely	NN	O	O
infiltrated	NN	O	O
after	NN	O	O
recruitment	NN	O	O
from	NN	O	O
the	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
.	NN	O	O

To	NN	O	O
ascertain	NN	O	O
this	NN	O	O
possibility	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
markers	NN	O	O
for	NN	O	O
B-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
proliferation	NN	O	O
in	NN	O	O
ILFs	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
C	NN	O	O
using	NN	O	O
surgically	NN	O	O
resected	NN	O	O
specimens	NN	O	O
,	NN	O	O
and	NN	O	O
compared	NN	O	O
them	NN	O	O
with	NN	O	O
specimens	NN	O	O
of	NN	O	O
perihepatic	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
by	NN	O	O
an	NN	O	O
immunohistochemical	NN	O	O
technique	NN	O	O
.	NN	O	O

Germinal	NN	O	O
center	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
formation	NN	O	O
in	NN	O	O
the	NN	O	O
ILFs	NN	O	O
was	NN	O	O
frequently	NN	O	O
found	NN	O	O
in	NN	O	O
HCV-positive	NN	O	O
cases	NN	O	O
.	NN	O	O

The	NN	O	O
distribution	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	O
M	NN	O	O
(	NN	O	O
IgM	NN	O	O
)	NN	O	O
-	NN	O	O
,	NN	O	O
IgD-	NN	O	O
,	NN	O	O
and	NN	O	O
IgG-positive	NN	O	O
cells	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
patterns	NN	O	O
of	NN	O	O
Ki-67	NN	O	O
,	NN	O	O
CD23	NN	O	O
,	NN	O	O
or	NN	O	O
bcl-2	NN	O	O
and	NN	O	O
bcl-6	NN	O	O
gene	NN	O	O
products	NN	O	O
in	NN	O	O
the	NN	O	O
follicles	NN	O	O
with	NN	O	O
GC	NN	O	O
formation	NN	O	O
in	NN	O	O
the	NN	O	O
liver	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
C	NN	O	O
were	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
activated	NN	O	O
,	NN	O	O
proliferated	NN	O	O
,	NN	O	O
and	NN	O	O
differentiated	NN	O	O
in	NN	O	O
the	NN	O	O
ILFs	NN	O	O
with	NN	O	O
GC	NN	O	O
formation	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
C	NN	O	O
.	NN	O	O

Oligoclonal	NN	O	O
expansion	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
livers	NN	O	O
with	NN	O	O
ILFs	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
an	NN	O	O
analysis	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
(	NN	O	I-DNA
IgH	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
rearrangement	NN	O	O
determined	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
ILFs	NN	O	O
with	NN	O	O
GC	NN	O	O
formation	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
frequently	NN	O	O
found	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
C	NN	O	O
,	NN	O	O
may	NN	O	O
functionally	NN	O	O
be	NN	O	O
the	NN	O	O
same	NN	O	O
as	NN	O	O
those	NN	O	O
found	NN	O	O
in	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
B-cell	NN	O	O
expansion	NN	O	O
and	NN	O	O
maturation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Downregulation	NN	O	O
of	NN	O	O
Wilms	NN	O	B-DNA
'	NN	O	I-DNA
tumor	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
WT1	NN	O	B-DNA
)	NN	O	O
is	NN	O	O
not	NN	O	O
a	NN	O	O
prerequisite	NN	O	O
for	NN	O	O
erythroid	NN	O	O
or	NN	O	O
megakaryocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
K562	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
Wilms	NN	O	B-DNA
'	NN	O	I-DNA
tumor	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
WT1	NN	O	B-DNA
)	NN	O	O
encodes	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
type	NN	O	I-protein
.	NN	O	O

A	NN	O	O
high	NN	O	O
expression	NN	O	O
of	NN	O	O
WT1	NN	O	B-DNA
has	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
a	NN	O	O
range	NN	O	O
of	NN	O	O
acute	NN	O	O
leukemias	NN	O	O
,	NN	O	O
and	NN	O	O
WT1	NN	O	B-DNA
is	NN	O	O
downregulated	NN	O	O
during	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
some	NN	O	O
leukemic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
WT1	NN	O	B-DNA
in	NN	O	O
some	NN	O	O
myeloid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
confers	NN	O	O
resistance	NN	O	O
to	NN	O	O
differentiation	NN	O	O
induction	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
high	NN	O	O
WT1	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
incompatible	NN	O	O
with	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
four	NN	O	O
different	NN	O	O
isoforms	NN	O	O
of	NN	O	O
WT1	NN	O	B-DNA
was	NN	O	O
constitutively	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
K562	NN	O	I-cell_line
.	NN	O	O

K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
express	NN	O	O
endogenous	NN	O	O
WT1	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
downregulated	NN	O	O
as	NN	O	O
a	NN	O	O
response	NN	O	O
to	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
along	NN	O	O
the	NN	O	O
erythroid	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
pathways	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
forced	NN	O	O
exogenous	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
four	NN	O	O
different	NN	O	O
isoforms	NN	O	O
of	NN	O	O
WT1	NN	O	B-DNA
in	NN	O	O
K562	NN	O	B-cell_line
does	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
differentiation	NN	O	O
response	NN	O	O
,	NN	O	O
as	NN	O	O
judged	NN	O	O
by	NN	O	O
accumulation	NN	O	O
of	NN	O	O
hemoglobin	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
hemin	NN	O	B-protein
or	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
megakaryocytic	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
markers	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
downregulation	NN	O	O
of	NN	O	O
WT1	NN	O	B-DNA
during	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
not	NN	O	O
a	NN	O	O
prerequisite	NN	O	O
for	NN	O	O
erythroid	NN	O	O
or	NN	O	O
megakaryocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Association	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-4	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
with	NN	O	O
p47phox	NN	O	O
,	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
the	NN	O	O
phagocyte	NN	O	B-protein
NADPH	NN	O	I-protein
oxidase	NN	O	I-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-4	NN	O	I-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

IL-4	NN	O	B-protein
conducts	NN	O	O
its	NN	O	O
biological	NN	O	O
activities	NN	O	O
through	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-4R	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

IL-4R	NN	O	B-protein
are	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
composed	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
and	NN	O	O
either	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-protein
gamma	NN	O	I-protein
chain	NN	O	I-protein
or	NN	O	O
the	NN	O	O
IL-13R	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
membrane-proximal	NN	O	B-protein
portion	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
IL-4R	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
hIL-4R	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
proliferation	NN	O	O
,	NN	O	O
generation	NN	O	O
of	NN	O	O
germline	NN	O	O
epsilon	NN	O	O
transcript	NN	O	O
,	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
,	NN	O	O
based	NN	O	O
on	NN	O	O
analyses	NN	O	O
of	NN	O	O
truncated	NN	O	B-protein
hIL-4R	NN	O	I-protein
alphas	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
p47phox	NN	O	O
,	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
the	NN	O	O
phagocyte	NN	O	B-protein
NADPH	NN	O	I-protein
oxidase	NN	O	I-protein
,	NN	O	O
binds	NN	O	O
to	NN	O	O
this	NN	O	O
portion	NN	O	O
by	NN	O	O
the	NN	O	O
two-hybrid	NN	O	O
system	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
p47phox	NN	O	O
with	NN	O	O
the	NN	O	O
hIL-4R	NN	O	B-protein
alpha	NN	O	I-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
derived	NN	O	O
from	NN	O	O
a	NN	O	O
normal	NN	O	O
donor	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
p47phox	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
,	NN	O	O
CD23	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
were	NN	O	O
not	NN	O	O
affected	NN	O	O
in	NN	O	O
p47phox-deficient	NN	O	O
patients	NN	O	O
,	NN	O	O
which	NN	O	O
raises	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
p47phox	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
other	NN	O	O
signaling	NN	O	O
activities	NN	O	O
as	NN	O	O
well	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
diesel	NN	O	O
organic	NN	O	O
extracts	NN	O	O
on	NN	O	O
chemokine	NN	O	B-protein
production	NN	O	O
by	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Polyaromatic	NN	O	O
hydrocarbons	NN	O	O
(	NN	O	O
PAHs	NN	O	O
)	NN	O	O
associated	NN	O	O
with	NN	O	O
diesel	NN	O	O
exhaust	NN	O	O
particles	NN	O	O
(	NN	O	O
DEPs	NN	O	O
)	NN	O	O
are	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
atmospheric	NN	O	O
urban	NN	O	O
pollution	NN	O	O
.	NN	O	O

Such	NN	O	O
compounds	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
favor	NN	O	O
IgE	NN	O	B-protein
production	NN	O	O
,	NN	O	O
bronchial	NN	O	O
hyperresponsiveness	NN	O	O
,	NN	O	O
and	NN	O	O
airway	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Chemokines	NN	O	O
are	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
chemotactic	NN	O	B-protein
cytokines	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
We	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
DEP-PAHs	NN	O	O
on	NN	O	O
the	NN	O	O
release	NN	O	O
and	NN	O	O
mRNA	NN	O	B-RNA
expression	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
MCP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
RANTES	NN	O	B-protein
by	NN	O	O
PBMCs	NN	O	B-cell_type
obtained	NN	O	O
from	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Protein	NN	O	O
production	NN	O	O
in	NN	O	O
supernatants	NN	O	O
was	NN	O	O
assessed	NN	O	O
by	NN	O	O
ELISA	NN	O	O
,	NN	O	O
and	NN	O	O
mRNA	NN	O	B-RNA
expression	NN	O	O
was	NN	O	O
evaluated	NN	O	O
by	NN	O	O
semiquantitative	NN	O	O
RT-PCR	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Secretion	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
and	NN	O	O
RANTES	NN	O	B-protein
increased	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
with	NN	O	O
increasing	NN	O	O
concentrations	NN	O	O
of	NN	O	O
DEP-PAHs	NN	O	O
(	NN	O	O
range	NN	O	O
,	NN	O	O
0.5	NN	O	O
ng	NN	O	O
to	NN	O	O
50	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
contrary	NN	O	O
,	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
was	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
,	NN	O	O
also	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Messenger	NN	O	B-RNA
RNA	NN	O	I-RNA
production	NN	O	O
coding	NN	O	O
for	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
RANTES	NN	O	B-protein
,	NN	O	O
and	NN	O	O
MCP-1	NN	O	B-protein
showed	NN	O	O
parallel	NN	O	O
variations	NN	O	O
to	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
the	NN	O	O
correspondent	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Effects	NN	O	O
of	NN	O	O
DEP-PAHs	NN	O	O
became	NN	O	O
significant	NN	O	O
at	NN	O	O
7	NN	O	O
hours	NN	O	O
and	NN	O	O
up	NN	O	O
to	NN	O	O
48	NN	O	O
hours	NN	O	O
time	NN	O	O
culture	NN	O	O
for	NN	O	O
MCP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
up	NN	O	O
to	NN	O	O
24	NN	O	O
hours	NN	O	O
time	NN	O	O
culture	NN	O	O
for	NN	O	O
IL-8	NN	O	B-protein
and	NN	O	O
RANTES	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
supernatants	NN	O	O
from	NN	O	O
DEP-PAH-activated	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
controls	NN	O	O
,	NN	O	O
exhibited	NN	O	O
a	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
chemotactic	NN	O	O
activity	NN	O	O
for	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
eosinophils	NN	O	B-cell_type
,	NN	O	O
which	NN	O	O
was	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
anti-IL-8	NN	O	B-protein
and	NN	O	O
anti-RANTES	NN	O	B-protein
neutralizing	NN	O	O
antibodies	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
chemokine	NN	O	B-protein
pathways	NN	O	O
are	NN	O	O
modulated	NN	O	O
by	NN	O	O
DEP-PAHs	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
,	NN	O	O
reinforcing	NN	O	O
the	NN	O	O
idea	NN	O	O
that	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
reactions	NN	O	O
might	NN	O	O
be	NN	O	O
affected	NN	O	O
by	NN	O	O
diesel	NN	O	O
exhaust	NN	O	O
emission	NN	O	O
.	NN	O	O

-DOCSTART-	O

20-Epi	NN	O	O
analogues	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
are	NN	O	O
highly	NN	O	O
potent	NN	O	O
inducers	NN	O	O
of	NN	O	O
DRIP	NN	O	B-protein
coactivator	NN	O	I-protein
complex	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D3	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

1	NN	O	O
,	NN	O	O
25-Dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
)	NN	O	O
plays	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
bone	NN	O	O
growth	NN	O	O
,	NN	O	O
mineralization	NN	O	O
,	NN	O	O
and	NN	O	O
intestinal	NN	O	O
calcium	NN	O	O
and	NN	O	O
phosphate	NN	O	O
absorption	NN	O	O
;	NN	O	O
it	NN	O	O
also	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
general	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Several	NN	O	O
new	NN	O	O
,	NN	O	O
clinically	NN	O	O
relevant	NN	O	O
compounds	NN	O	O
dissociate	NN	O	O
antiproliferative	NN	O	O
and	NN	O	O
calcemic	NN	O	O
activities	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
this	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
clearly	NN	O	O
elucidated	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
tested	NN	O	O
whether	NN	O	O
the	NN	O	O
potency	NN	O	O
of	NN	O	O
one	NN	O	O
class	NN	O	O
of	NN	O	O
compounds	NN	O	O
,	NN	O	O
20-epi	NN	O	O
analogues	NN	O	O
,	NN	O	O
to	NN	O	O
induce	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
is	NN	O	O
because	NN	O	O
of	NN	O	O
direct	NN	O	O
molecular	NN	O	O
effects	NN	O	O
on	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
two	NN	O	O
20-epi	NN	O	O
analogues	NN	O	O
,	NN	O	O
MC1627	NN	O	O
and	NN	O	O
MC1288	NN	O	O
,	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
and	NN	O	O
transcription	NN	O	O
of	NN	O	O
p21	NN	O	B-DNA
(	NN	O	O
Waf1	NN	O	B-DNA
,	NN	O	O
Cip1	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
a	NN	O	O
key	NN	O	O
VDR	NN	O	B-DNA
target	NN	O	I-DNA
gene	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
,	NN	O	O
at	NN	O	O
a	NN	O	O
concentration	NN	O	O
100-fold	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
.	NN	O	O

We	NN	O	O
compared	NN	O	O
this	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
analogue	NN	O	O
effects	NN	O	O
on	NN	O	O
VDR	NN	O	B-protein
interacting	NN	O	O
proteins	NN	O	O
:	NN	O	O
RXR	NN	O	B-protein
,	NN	O	O
GRIP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
DRIP205	NN	O	B-protein
,	NN	O	O
a	NN	O	O
subunit	NN	O	O
of	NN	O	O
the	NN	O	O
DRIP	NN	O	B-protein
coactivator	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Compared	NN	O	O
with	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
with	NN	O	O
RXR	NN	O	B-protein
or	NN	O	O
GRIP-1	NN	O	B-protein
,	NN	O	O
the	NN	O	O
differentiation	NN	O	O
dose-response	NN	O	O
most	NN	O	O
closely	NN	O	O
correlated	NN	O	O
to	NN	O	O
the	NN	O	O
ligand-dependent	NN	O	O
recruitment	NN	O	O
of	NN	O	O
the	NN	O	O
DRIP	NN	O	B-protein
coactivator	NN	O	I-protein
complex	NN	O	I-protein
to	NN	O	O
VDR	NN	O	B-protein
and	NN	O	O
to	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
in	NN	O	O
a	NN	O	O
cell-free	NN	O	O
system	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
compelling	NN	O	O
links	NN	O	O
between	NN	O	O
the	NN	O	O
efficiency	NN	O	O
of	NN	O	O
the	NN	O	O
20-epi	NN	O	O
analogue	NN	O	O
in	NN	O	O
inducing	NN	O	O
VDR	NN	O	B-protein
/DRIP	NN	O	O
interactions	NN	O	O
,	NN	O	O
transactivation	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
enhanced	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

SLP-76	NN	O	B-protein
and	NN	O	O
Vav	NN	O	B-protein
function	NN	O	O
in	NN	O	O
separate	NN	O	O
,	NN	O	O
but	NN	O	O
overlapping	NN	O	O
pathways	NN	O	O
to	NN	O	O
augment	NN	O	O
interleukin-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

SLP-76	NN	O	B-protein
and	NN	O	O
Vav	NN	O	B-protein
,	NN	O	O
two	NN	O	O
hematopoietic	NN	O	B-protein
cell	NN	O	I-protein
specific	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
are	NN	O	O
critical	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
and	NN	O	O
activation	NN	O	O
.	NN	O	O

Following	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
stimulation	NN	O	O
,	NN	O	O
SLP-76	NN	O	B-protein
and	NN	O	O
Vav	NN	O	B-protein
both	NN	O	O
undergo	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
associate	NN	O	O
with	NN	O	O
each	NN	O	O
other	NN	O	O
via	NN	O	O
the	NN	O	O
SH2	NN	O	O
domain	NN	O	O
of	NN	O	O
Vav	NN	O	B-protein
and	NN	O	O
phosphorylated	NN	O	O
tyrosines	NN	O	O
of	NN	O	O
SLP-76	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
SLP-76	NN	O	B-protein
and	NN	O	O
Vav	NN	O	B-protein
have	NN	O	O
a	NN	O	O
synergistic	NN	O	O
effect	NN	O	O
on	NN	O	O
interleukin	NN	O	B-DNA
(	NN	O	I-DNA
IL	NN	O	I-DNA
)	NN	O	I-DNA
-2	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
two	NN	O	O
tyrosines	NN	O	O
,	NN	O	O
Tyr-113	NN	O	O
and	NN	O	O
Tyr-128	NN	O	O
,	NN	O	O
of	NN	O	O
SLP-76	NN	O	B-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
its	NN	O	O
binding	NN	O	O
to	NN	O	O
Vav	NN	O	B-protein
,	NN	O	O
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
intact	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
we	NN	O	O
find	NN	O	O
also	NN	O	O
that	NN	O	O
the	NN	O	O
interaction	NN	O	O
between	NN	O	O
SLP-76	NN	O	B-protein
and	NN	O	O
Vav	NN	O	B-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
their	NN	O	O
cooperation	NN	O	O
in	NN	O	O
augmenting	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
,	NN	O	O
as	NN	O	O
the	NN	O	O
two	NN	O	O
molecules	NN	O	O
appear	NN	O	O
to	NN	O	O
function	NN	O	O
in	NN	O	O
different	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
upstream	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
SLP-76	NN	O	B-protein
in	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
potentiates	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
both	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Vav	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
enhanced	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
activity	NN	O	O
without	NN	O	O
affecting	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Vav	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
SLP-76	NN	O	B-protein
,	NN	O	O
augments	NN	O	O
CD28-induced	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
synergy	NN	O	O
between	NN	O	O
SLP-76	NN	O	B-protein
and	NN	O	O
Vav	NN	O	B-protein
in	NN	O	O
regulating	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
reflects	NN	O	O
the	NN	O	O
cooperation	NN	O	O
between	NN	O	O
different	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

Rel/NF-kappaB	NN	O	B-protein
can	NN	O	O
trigger	NN	O	O
the	NN	O	O
Notch	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
by	NN	O	O
inducing	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Jagged1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
ligand	NN	O	O
for	NN	O	O
Notch	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Jagged1	NN	O	B-protein
belongs	NN	O	O
to	NN	O	O
the	NN	O	O
DSL	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
ligands	NN	O	I-protein
for	NN	O	O
Notch	NN	O	B-protein
receptors	NN	O	I-protein
that	NN	O	O
control	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
various	NN	O	O
cell	NN	O	B-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
its	NN	O	O
expression	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
Jagged1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
Rel/NF-kappaB-responsive	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	O
RelA	NN	O	B-protein
induced	NN	O	O
jagged1	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
mutant	NN	O	O
defective	NN	O	O
for	NN	O	O
transactivation	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
jagged1	NN	O	B-protein
transcripts	NN	O	I-protein
were	NN	O	O
also	NN	O	O
upregulated	NN	O	O
by	NN	O	O
endogenous	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
this	NN	O	O
effect	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
dominant	NN	O	O
mutant	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
,	NN	O	O
a	NN	O	O
physiological	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
Jagged1	NN	O	B-protein
in	NN	O	O
c-Rel-expressing	NN	O	B-cell_line
cell	NN	O	I-cell_line
monolayers	NN	O	I-cell_line
led	NN	O	O
to	NN	O	O
a	NN	O	O
functional	NN	O	O
interaction	NN	O	O
with	NN	O	O
lymphocytes	NN	O	B-cell_type
expressing	NN	O	O
the	NN	O	O
Notch1/TAN-1	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

This	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
signaling	NN	O	O
downstream	NN	O	O
of	NN	O	O
Notch	NN	O	O
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
HES-1	NN	O	B-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
co-cultivated	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
of	NN	O	O
CD23	NN	O	B-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
co-cultivated	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
its	NN	O	O
Rel/NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
induction	NN	O	O
,	NN	O	O
Jagged1	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
highly	NN	O	O
expressed	NN	O	O
in	NN	O	O
splenic	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
where	NN	O	O
c-Rel	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
constitutively	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
c-Rel	NN	O	B-protein
can	NN	O	O
trigger	NN	O	O
the	NN	O	O
Notch	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
in	NN	O	O
neighboring	NN	O	O
cells	NN	O	O
by	NN	O	O
inducing	NN	O	O
jagged1	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
Jagged1	NN	O	B-protein
in	NN	O	O
B-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
differentiation	NN	O	O
or	NN	O	O
function	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
also	NN	O	O
highlight	NN	O	O
the	NN	O	O
potential	NN	O	O
for	NN	O	O
an	NN	O	O
interplay	NN	O	O
between	NN	O	O
the	NN	O	O
Notch	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

-DOCSTART-	O

Bacterial	NN	O	O
peptidoglycan	NN	O	O
induces	NN	O	O
CD14	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
CREB/ATF	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Peptidoglycan	NN	O	O
(	NN	O	O
PGN	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
major	NN	O	O
cell	NN	O	O
wall	NN	O	O
component	NN	O	O
of	NN	O	O
Gram-positive	NN	O	O
bacteria	NN	O	O
,	NN	O	O
induces	NN	O	O
secretion	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
macrophages	NN	O	B-cell_type
through	NN	O	O
CD14	NN	O	B-protein
,	NN	O	O
the	NN	O	O
pattern	NN	O	O
recognition	NN	O	O
receptor	NN	O	O
that	NN	O	O
binds	NN	O	O
lipopolysaccharide	NN	O	O
and	NN	O	O
other	NN	O	O
microbial	NN	O	O
products	NN	O	O
.	NN	O	O

To	NN	O	O
begin	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
regulate	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
wanted	NN	O	O
to	NN	O	O
determine	NN	O	O
which	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
activated	NN	O	O
by	NN	O	O
PGN	NN	O	O
in	NN	O	O
mouse	NN	O	B-cell_line
RAW264.7	NN	O	I-cell_line
and	NN	O	O
human	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
macrophage	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
:	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
PGN	NN	O	O
induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
ATF-1	NN	O	B-protein
and	NN	O	O
CREB	NN	O	B-protein
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
ATF-1	NN	O	B-protein
and	NN	O	O
CREB	NN	O	B-protein
bound	NN	O	O
DNA	NN	O	O
as	NN	O	O
a	NN	O	O
dimer	NN	O	O
and	NN	O	O
induced	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
CRE	NN	O	B-DNA
reporter	NN	O	I-DNA
plasmid	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
dominant	NN	O	O
negative	NN	O	O
CREB	NN	O	B-protein
and	NN	O	O
ATF-1	NN	O	B-protein
;	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
PGN	NN	O	O
induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
of	NN	O	O
JunB	NN	O	B-protein
and	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
reporter	NN	O	I-DNA
plasmid	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
dominant	NN	O	O
negative	NN	O	O
c-Fos	NN	O	B-protein
;	NN	O	O
and	NN	O	O
(	NN	O	O
iv	NN	O	O
)	NN	O	O
PGN-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
CREB/ATF	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
was	NN	O	O
mediated	NN	O	O
through	NN	O	O
CD14	NN	O	B-protein
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
study	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
activation	NN	O	O
of	NN	O	O
CREB/ATF	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
by	NN	O	O
PGN	NN	O	O
or	NN	O	O
by	NN	O	O
any	NN	O	O
other	NN	O	O
component	NN	O	O
of	NN	O	O
Gram-positive	NN	O	O
bacteria	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
are	NN	O	O
down-regulated	NN	O	O
in	NN	O	O
inflamed	NN	O	O
colonic	NN	O	O
mucosa	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
inflammatory	NN	O	O
bowel	NN	O	O
disease	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Growing	NN	O	O
evidence	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
and	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
system	NN	O	O
are	NN	O	O
linked	NN	O	O
by	NN	O	O
several	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
for	NN	O	O
example	NN	O	O
intracellular	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
hGR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
are	NN	O	O
the	NN	O	O
standard	NN	O	O
treatment	NN	O	O
of	NN	O	O
acute	NN	O	O
attacks	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
bowel	NN	O	O
disease	NN	O	O
(	NN	O	O
IBD	NN	O	O
)	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
to	NN	O	O
hGR	NN	O	B-protein
down-regulates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-DNA
genes	NN	O	I-DNA
that	NN	O	O
can	NN	O	O
propagate	NN	O	O
IBD	NN	O	O
.	NN	O	O

PATIENTS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
IBD	NN	O	O
patients	NN	O	O
were	NN	O	O
either	NN	O	O
treated	NN	O	O
with	NN	O	O
5-60	NN	O	O
mg	NN	O	O
of	NN	O	O
prednisolone	NN	O	O
for	NN	O	O
more	NN	O	O
than	NN	O	O
1	NN	O	O
week	NN	O	O
or	NN	O	O
were	NN	O	O
without	NN	O	O
glucocorticoid	NN	O	O
treatment	NN	O	O
for	NN	O	O
more	NN	O	O
than	NN	O	O
4	NN	O	O
weeks	NN	O	O
.	NN	O	O

hGR	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
determined	NN	O	O
from	NN	O	O
isolated	NN	O	O
cytosol	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
or	NN	O	O
mucosal	NN	O	O
biopsies	NN	O	O
using	NN	O	O
a	NN	O	O
radioassay	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-dexamethasone	NN	O	O
.	NN	O	O

Interleukin	NN	O	O
(	NN	O	O
IL	NN	O	O
)	NN	O	O
6	NN	O	O
levels	NN	O	O
were	NN	O	O
determined	NN	O	O
by	NN	O	O
enzyme-linked	NN	O	O
immunosorbent	NN	O	O
assay	NN	O	O
(	NN	O	O
ELISA	NN	O	O
)	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
The	NN	O	O
systemic	NN	O	O
(	NN	O	O
PBMC	NN	O	O
)	NN	O	O
hGR	NN	O	B-protein
levels	NN	O	O
of	NN	O	O
corticosteroid-treated	NN	O	O
IBD	NN	O	O
patients	NN	O	O
were	NN	O	O
significantly	NN	O	O
lower	NN	O	O
than	NN	O	O
those	NN	O	O
of	NN	O	O
control	NN	O	O
subjects	NN	O	O
(	NN	O	O
59.6	NN	O	O
+/-	NN	O	O
57.1	NN	O	O
dpm	NN	O	O
mg-1	NN	O	O
cytosol	NN	O	B-protein
protein	NN	O	I-protein
vs	NN	O	O
.	NN	O	O

227.0	NN	O	O
+/-	NN	O	O
90.8	NN	O	O
dpm	NN	O	O
mg-1	NN	O	O
cytosol	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
P	NN	O	O
=	NN	O	O
0.007	NN	O	O
)	NN	O	O
and	NN	O	O
IBD	NN	O	O
patients	NN	O	O
not	NN	O	O
receiving	NN	O	O
glucocorticoid	NN	O	O
treatment	NN	O	O
(	NN	O	O
179.7	NN	O	O
+/-	NN	O	O
171.3	NN	O	O
dpm	NN	O	O
mg-1	NN	O	O
cytosol	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
P	NN	O	O
=	NN	O	O
0.002	NN	O	O
)	NN	O	O
.	NN	O	O

Systemic	NN	O	O
hGR	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
untreated	NN	O	O
IBD	NN	O	O
patients	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
significantly	NN	O	O
from	NN	O	O
those	NN	O	O
in	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

In	NN	O	O
patients	NN	O	O
with	NN	O	O
connective	NN	O	O
tissue	NN	O	O
diseases	NN	O	O
,	NN	O	O
systemic	NN	O	O
hGR	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
also	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
decreased	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
treatment	NN	O	O
.	NN	O	O

Systemic	NN	O	O
hGR	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Crohn	NN	O	O
's	NN	O	O
disease	NN	O	O
(	NN	O	O
CD	NN	O	O
)	NN	O	O
treated	NN	O	O
with	NN	O	O
steroids	NN	O	O
(	NN	O	O
66.6	NN	O	O
+/-	NN	O	O
61.0	NN	O	O
dpm	NN	O	O
mg-1	NN	O	O
cytosol	NN	O	B-protein
protein	NN	O	I-protein
)	NN	O	O
were	NN	O	O
not	NN	O	O
different	NN	O	O
from	NN	O	O
those	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
ulcerative	NN	O	O
colitis	NN	O	O
(	NN	O	O
UC	NN	O	O
)	NN	O	O
(	NN	O	O
56.1	NN	O	O
+/-	NN	O	O
51.6	NN	O	O
dpm	NN	O	O
mg-1	NN	O	O
cytosol	NN	O	B-protein
protein	NN	O	I-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
mucosal	NN	O	O
hGR	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
in	NN	O	O
both	NN	O	O
steroid-treated	NN	O	O
(	NN	O	O
18.0	NN	O	O
+/-	NN	O	O
15.5	NN	O	O
)	NN	O	O
and	NN	O	O
not	NN	O	O
steroid-treated	NN	O	O
(	NN	O	O
37.8	NN	O	O
+/-	NN	O	O
30.5	NN	O	O
)	NN	O	O
patients	NN	O	O
compared	NN	O	O
with	NN	O	O
control	NN	O	O
subjects	NN	O	O
(	NN	O	O
125.6	NN	O	O
+/-	NN	O	O
97.1	NN	O	O
;	NN	O	O
P	NN	O	O
=	NN	O	O
0.00009	NN	O	O
and	NN	O	O
P	NN	O	O
=	NN	O	O
0.0008	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

IL-6	NN	O	O
levels	NN	O	O
in	NN	O	O
all	NN	O	O
IBD	NN	O	O
groups	NN	O	O
with	NN	O	O
and	NN	O	O
without	NN	O	O
steroids	NN	O	O
were	NN	O	O
significantly	NN	O	O
different	NN	O	O
from	NN	O	O
those	NN	O	O
in	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
In	NN	O	O
IBD	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
difference	NN	O	O
in	NN	O	O
systemic	NN	O	O
hGR	NN	O	B-protein
levels	NN	O	O
between	NN	O	O
not	NN	O	O
steroid-treated	NN	O	O
patients	NN	O	O
and	NN	O	O
control	NN	O	O
subjects	NN	O	O
,	NN	O	O
in	NN	O	O
spite	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
activity	NN	O	O
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Mucosal	NN	O	O
hGR	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
decreased	NN	O	O
independently	NN	O	O
of	NN	O	O
treatment	NN	O	O
,	NN	O	O
probably	NN	O	O
leading	NN	O	O
to	NN	O	O
a	NN	O	O
decreased	NN	O	O
protection	NN	O	O
against	NN	O	O
NF-kappaB	NN	O	B-protein
action	NN	O	O
in	NN	O	O
the	NN	O	O
intestinal	NN	O	O
mucosa	NN	O	O
.	NN	O	O

-DOCSTART-	O

ICSAT	NN	O	B-protein
overexpression	NN	O	O
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
cause	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
or	NN	O	O
multiple	NN	O	O
myeloma	NN	O	O
.	NN	O	O

ICSAT	NN	O	B-protein
(	NN	O	O
Interferon	NN	O	B-protein
Consensus	NN	O	I-protein
Sequence	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
for	NN	O	I-protein
Activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
lymphocyte-specific	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
interferon	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
IRF	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	O
factors	NN	O	O
,	NN	O	O
originally	NN	O	O
identified	NN	O	O
through	NN	O	O
Southwestern	NN	O	O
screening	NN	O	O
of	NN	O	O
the	NN	O	O
ATL	NN	O	B-DNA
(	NN	O	I-DNA
Adult	NN	O	I-DNA
T-cell	NN	O	I-DNA
leukemia	NN	O	I-DNA
)	NN	O	I-DNA
-16T	NN	O	I-DNA
expression	NN	O	I-DNA
library	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
created	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
overexpressing	NN	O	O
ICSAT	NN	O	B-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Although	NN	O	O
spontaneous	NN	O	O
tumorigenesis	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
with	NN	O	O
Concanavalin	NN	O	O
A	NN	O	O
stimulation	NN	O	O
was	NN	O	O
significantly	NN	O	O
increased	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
overexpressing	NN	O	O
ICSAT	NN	O	B-protein
.	NN	O	O

ICSAT	NN	O	B-protein
overexpression	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
seems	NN	O	O
insufficient	NN	O	O
for	NN	O	O
the	NN	O	O
leukemogenesis	NN	O	O
of	NN	O	O
ATL	NN	O	O
or	NN	O	O
multiple	NN	O	O
myeloma	NN	O	O
(	NN	O	O
MM	NN	O	O
)	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
may	NN	O	O
regulate	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
its	NN	O	O
overexpression	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
leukemogenesis	NN	O	O
via	NN	O	O
controlling	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Altered	NN	O	O
memory	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
early	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
.	NN	O	O

The	NN	O	O
chronic	NN	O	O
immune	NN	O	O
response	NN	O	O
in	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
might	NN	O	O
be	NN	O	O
driven	NN	O	O
by	NN	O	O
activated	NN	O	O
Th1	NN	O	O
cells	NN	O	O
without	NN	O	O
sufficient	NN	O	O
Th2	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
to	NN	O	O
down-modulate	NN	O	O
inflammation	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
whether	NN	O	O
disordered	NN	O	O
memory	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
typical	NN	O	O
Th1-dominated	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
in	NN	O	O
RA	NN	O	O
we	NN	O	O
investigated	NN	O	O
differentiation	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
memory	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
early	NN	O	O
(	NN	O	O
6	NN	O	O
wk	NN	O	O
to	NN	O	O
12	NN	O	O
mo	NN	O	O
)	NN	O	O
untreated	NN	O	O
RA	NN	O	O
and	NN	O	O
in	NN	O	O
age-	NN	O	O
and	NN	O	O
sex-matched	NN	O	O
healthy	NN	O	O
controls	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

No	NN	O	O
difference	NN	O	O
in	NN	O	O
cytokine	NN	O	B-protein
secretion	NN	O	O
profiles	NN	O	O
of	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
memory	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
detected	NN	O	O
between	NN	O	O
patients	NN	O	O
and	NN	O	O
controls	NN	O	O
.	NN	O	O

A	NN	O	O
cell	NN	O	B-cell_line
culture	NN	O	I-cell_line
system	NN	O	I-cell_line
was	NN	O	O
then	NN	O	O
employed	NN	O	O
that	NN	O	O
permitted	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
Th	NN	O	B-protein
effectors	NN	O	I-protein
from	NN	O	O
resting	NN	O	B-cell_type
memory	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
short	NN	O	O
term	NN	O	O
priming	NN	O	O
.	NN	O	O

Marked	NN	O	O
differences	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
priming	NN	O	O
.	NN	O	O

Th2	NN	O	B-cell_line
cells	NN	O	I-cell_line
could	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
all	NN	O	O
healthy	NN	O	O
controls	NN	O	O
by	NN	O	O
priming	NN	O	O
with	NN	O	O
anti-CD28	NN	O	B-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
ligation	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
priming	NN	O	O
under	NN	O	O
those	NN	O	O
conditions	NN	O	O
resulted	NN	O	O
in	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
in	NN	O	O
only	NN	O	O
9	NN	O	O
of	NN	O	O
24	NN	O	O
RA	NN	O	O
patients	NN	O	O
.	NN	O	O

Exogenous	NN	O	O
IL-4	NN	O	B-protein
could	NN	O	O
overcome	NN	O	O
the	NN	O	O
apparent	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
defect	NN	O	O
in	NN	O	O
seven	NN	O	O
patients	NN	O	O
but	NN	O	O
was	NN	O	O
without	NN	O	O
effect	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
eight	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
patients	NN	O	O
a	NN	O	O
marked	NN	O	O
decrease	NN	O	O
in	NN	O	O
IL-2-producing	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
in	NN	O	O
well-differentiated	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
produced	NN	O	O
IFN-gamma	NN	O	B-protein
but	NN	O	O
not	NN	O	O
IL-2	NN	O	B-protein
were	NN	O	O
evident	NN	O	O
after	NN	O	O
priming	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
and	NN	O	O
anti-CD28	NN	O	B-protein
.	NN	O	O

The	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
CD4+	NN	O	B-cell_type
memory	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
early	NN	O	O
untreated	NN	O	O
RA	NN	O	O
manifest	NN	O	O
an	NN	O	O
intrinsic	NN	O	O
abnormality	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
differentiate	NN	O	O
into	NN	O	O
specific	NN	O	O
cytokine-producing	NN	O	B-cell_type
effector	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
might	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
characteristic	NN	O	O
Th1-dominated	NN	O	O
chronic	NN	O	O
(	NN	O	O
auto	NN	O	O
)	NN	O	O
immune	NN	O	O
inflammation	NN	O	O
in	NN	O	O
RA	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
intracellular	NN	O	O
parasite	NN	O	O
Theileria	NN	O	O
parva	NN	O	O
protects	NN	O	O
infected	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Parasites	NN	O	O
have	NN	O	O
evolved	NN	O	O
a	NN	O	O
plethora	NN	O	O
of	NN	O	O
strategies	NN	O	O
to	NN	O	O
ensure	NN	O	O
their	NN	O	O
survival	NN	O	O
.	NN	O	O

The	NN	O	O
intracellular	NN	O	O
parasite	NN	O	O
Theileria	NN	O	O
parva	NN	O	O
secures	NN	O	O
its	NN	O	O
propagation	NN	O	O
and	NN	O	O
spreads	NN	O	O
through	NN	O	O
the	NN	O	O
infected	NN	O	O
animal	NN	O	O
by	NN	O	O
infecting	NN	O	O
and	NN	O	O
transforming	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
inducing	NN	O	O
their	NN	O	O
continuous	NN	O	O
proliferation	NN	O	O
and	NN	O	O
rendering	NN	O	O
them	NN	O	O
metastatic	NN	O	O
.	NN	O	O

In	NN	O	O
previous	NN	O	O
work	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
parasite	NN	O	O
induces	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
,	NN	O	O
by	NN	O	O
inducing	NN	O	O
the	NN	O	O
constitutive	NN	O	O
degradation	NN	O	O
of	NN	O	O
its	NN	O	O
cytoplasmic	NN	O	B-protein
inhibitors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
biological	NN	O	O
significance	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
T.	NN	O	B-cell_type
parva-infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
however	NN	O	O
,	NN	O	O
has	NN	O	O
not	NN	O	O
yet	NN	O	O
been	NN	O	O
defined	NN	O	O
.	NN	O	O

Cells	NN	O	O
that	NN	O	O
have	NN	O	O
been	NN	O	O
transformed	NN	O	O
by	NN	O	O
viruses	NN	O	O
or	NN	O	O
oncogenes	NN	O	B-DNA
can	NN	O	O
persist	NN	O	O
only	NN	O	O
if	NN	O	O
they	NN	O	O
manage	NN	O	O
to	NN	O	O
avoid	NN	O	O
destruction	NN	O	O
by	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
are	NN	O	O
activated	NN	O	O
on	NN	O	O
transformation	NN	O	O
and	NN	O	O
that	NN	O	O
contribute	NN	O	O
to	NN	O	O
maintain	NN	O	O
cellular	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
parasite-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
T.	NN	O	B-cell_type
parva-transformed	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
conveying	NN	O	O
protection	NN	O	O
against	NN	O	O
an	NN	O	O
apoptotic	NN	O	O
signal	NN	O	O
that	NN	O	O
accompanies	NN	O	O
parasite-mediated	NN	O	O
transformation	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
mutant	NN	O	I-protein
forms	NN	O	O
of	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
pathway	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
IkappaBalpha	NN	O	B-protein
or	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
prompt	NN	O	O
rapid	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
T.	NN	O	B-cell_type
parva-transformed	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
offer	NN	O	O
important	NN	O	O
insights	NN	O	O
into	NN	O	O
parasite	NN	O	O
survival	NN	O	O
strategies	NN	O	O
and	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
parasite-induced	NN	O	O
constitutive	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
an	NN	O	O
essential	NN	O	O
step	NN	O	O
in	NN	O	O
maintaining	NN	O	O
the	NN	O	O
transformed	NN	O	O
phenotype	NN	O	O
of	NN	O	O
the	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
polymorphism	NN	O	O
that	NN	O	O
affects	NN	O	O
OCT-1	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
is	NN	O	O
associated	NN	O	O
with	NN	O	O
severe	NN	O	O
malaria	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Genetic	NN	O	O
variation	NN	O	O
in	NN	O	O
cytokine	NN	O	B-DNA
promoter	NN	O	I-DNA
regions	NN	O	I-DNA
is	NN	O	O
postulated	NN	O	O
to	NN	O	O
influence	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
infection	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
such	NN	O	O
polymorphisms	NN	O	O
might	NN	O	O
affect	NN	O	O
gene	NN	O	O
regulation	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

Through	NN	O	O
systematic	NN	O	O
DNA	NN	O	O
footprinting	NN	O	O
of	NN	O	O
the	NN	O	O
TNF	NN	O	O
(	NN	O	O
encoding	NN	O	O
tumour	NN	O	O
necrosis	NN	O	O
factor	NN	O	O
,	NN	O	O
TNF	NN	O	O
)	NN	O	O
promoter	NN	O	O
region	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
single	NN	O	O
nucleotide	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SNP	NN	O	O
)	NN	O	O
that	NN	O	O
causes	NN	O	O
the	NN	O	O
helix-turn-helix	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
OCT-1	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
a	NN	O	O
novel	NN	O	O
region	NN	O	O
of	NN	O	O
complex	NN	O	O
protein-DNA	NN	O	O
interactions	NN	O	O
and	NN	O	O
alters	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
OCT-1-binding	NN	O	O
genotype	NN	O	O
,	NN	O	O
found	NN	O	O
in	NN	O	O
approximately	NN	O	O
5	NN	O	O
%	NN	O	O
of	NN	O	O
Africans	NN	O	O
,	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
fourfold	NN	O	O
increased	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
cerebral	NN	O	O
malaria	NN	O	O
in	NN	O	O
large	NN	O	O
case-control	NN	O	O
studies	NN	O	O
of	NN	O	O
West	NN	O	O
African	NN	O	O
and	NN	O	O
East	NN	O	O
African	NN	O	O
populations	NN	O	O
,	NN	O	O
after	NN	O	O
correction	NN	O	O
for	NN	O	O
other	NN	O	O
known	NN	O	O
TNF	NN	O	O
polymorphisms	NN	O	O
and	NN	O	O
linked	NN	O	O
HLA	NN	O	B-DNA
alleles	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

UV-induced	NN	O	O
CYP1A1	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	O
cells	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
tryptophan	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
cytochrome	NN	O	B-protein
P-4501A1	NN	O	I-protein
(	NN	O	O
CYP1A1	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
by	NN	O	O
UV	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
earlier	NN	O	O
in	NN	O	O
animal	NN	O	O
studies	NN	O	O
via	NN	O	O
a	NN	O	O
mechanism	NN	O	O
that	NN	O	O
has	NN	O	O
not	NN	O	O
yet	NN	O	O
been	NN	O	O
resolved	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
data	NN	O	O
have	NN	O	O
indicated	NN	O	O
that	NN	O	O
formylated	NN	O	O
indolocarbazoles	NN	O	O
which	NN	O	O
are	NN	O	O
formed	NN	O	O
by	NN	O	O
UV	NN	O	O
irradiation	NN	O	O
of	NN	O	O
tryptophan	NN	O	O
solutions	NN	O	O
are	NN	O	O
very	NN	O	O
potent	NN	O	O
Ah-receptor	NN	O	O
agonists	NN	O	O
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
UV	NN	O	O
light	NN	O	O
on	NN	O	O
cytochrome	NN	O	B-protein
P4501A1	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
CYP1A1	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
UV	NN	O	O
irradiation	NN	O	O
of	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
keratinocytes	NN	O	I-cell_line
(	NN	O	O
HaCaT	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
mouse	NN	O	B-cell_line
Hepa-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
cells	NN	O	O
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
UV	NN	O	O
light	NN	O	O
delivered	NN	O	O
by	NN	O	O
a	NN	O	O
bank	NN	O	O
of	NN	O	O
6	NN	O	O
Philips	NN	O	O
TL20/12RS	NN	O	O
sun	NN	O	O
lamps	NN	O	O
emitting	NN	O	O
primarily	NN	O	O
in	NN	O	O
the	NN	O	O
UVB	NN	O	O
range	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
and	NN	O	O
presence	NN	O	O
of	NN	O	O
tryptophan	NN	O	O
.	NN	O	O

A	NN	O	O
semiquantitative	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
-linked	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
was	NN	O	O
used	NN	O	O
for	NN	O	O
analysis	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
treated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
CYP1A1	NN	O	B-RNA
mRNA	NN	O	I-RNA
level	NN	O	O
induced	NN	O	O
by	NN	O	O
UV	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
tryptophan	NN	O	O
was	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
induced	NN	O	O
by	NN	O	O
UV	NN	O	O
alone	NN	O	O
in	NN	O	O
both	NN	O	O
HaCaT	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
lymphocytes	NN	O	B-cell_type
after	NN	O	O
3	NN	O	O
h	NN	O	O
of	NN	O	O
incubation	NN	O	O
post-UV	NN	O	O
irradiation	NN	O	O
.	NN	O	O

To	NN	O	O
find	NN	O	O
out	NN	O	O
if	NN	O	O
the	NN	O	O
induction	NN	O	O
by	NN	O	O
UV	NN	O	O
light	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
an	NN	O	O
Ah	NN	O	O
receptor	NN	O	O
ligand	NN	O	O
,	NN	O	O
Hepa-1	NN	O	B-cell_line
wild-type	NN	O	I-cell_line
and	NN	O	I-cell_line
Ah	NN	O	I-cell_line
receptor	NN	O	I-cell_line
deficient	NN	O	I-cell_line
c12	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
applied	NN	O	O
.	NN	O	O

Wild-type	NN	O	B-cell_line
(	NN	O	I-cell_line
wt	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
inducible	NN	O	O
either	NN	O	O
by	NN	O	O
the	NN	O	O
tryptophan	NN	O	O
photoproduct	NN	O	O
6-formylindolo	NN	O	O
[	NN	O	O
3	NN	O	O
,	NN	O	O
2-b	NN	O	O
]	NN	O	O
carbazole	NN	O	O
(	NN	O	O
FICZ	NN	O	O
)	NN	O	O
or	NN	O	O
by	NN	O	O
UV-irradiation	NN	O	O
but	NN	O	O
very	NN	O	O
low	NN	O	O
or	NN	O	O
undetectable	NN	O	O
levels	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
c12	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
shows	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
FICZ	NN	O	O
and	NN	O	O
UV	NN	O	O
is	NN	O	O
Ah	NN	O	B-protein
receptor	NN	O	I-protein
dependent	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
UV-induced	NN	O	O
CYP1A1	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
an	NN	O	O
Ah	NN	O	B-protein
receptor	NN	O	I-protein
ligand	NN	O	O
formed	NN	O	O
from	NN	O	O
tryptophan	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
photoproducts	NN	O	O
of	NN	O	O
tryptophan	NN	O	O
are	NN	O	O
suggested	NN	O	O
to	NN	O	O
be	NN	O	O
mediators	NN	O	O
of	NN	O	O
light	NN	O	O
via	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
Ah	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
as	NN	O	O
such	NN	O	O
also	NN	O	O
could	NN	O	O
have	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
light-regulated	NN	O	O
biological	NN	O	O
rhythms	NN	O	O
.	NN	O	O

-DOCSTART-	O

Repression	NN	O	O
by	NN	O	O
Ikaros	NN	O	B-protein
and	NN	O	O
Aiolos	NN	O	B-protein
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
histone	NN	O	B-protein
deacetylase	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
lymphoid	NN	O	B-protein
lineage-determining	NN	O	I-protein
factors	NN	O	I-protein
Ikaros	NN	O	B-protein
and	NN	O	O
Aiolos	NN	O	B-protein
can	NN	O	O
function	NN	O	O
as	NN	O	O
strong	NN	O	B-protein
transcriptional	NN	O	I-protein
repressors	NN	O	I-protein
.	NN	O	O

This	NN	O	O
function	NN	O	O
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
two	NN	O	O
repression	NN	O	B-protein
domains	NN	O	I-protein
and	NN	O	O
is	NN	O	O
dependent	NN	O	O
upon	NN	O	O
the	NN	O	O
promoter	NN	O	O
context	NN	O	O
and	NN	O	O
cell	NN	O	O
type	NN	O	O
.	NN	O	O

Repression	NN	O	O
by	NN	O	O
Ikaros	NN	O	B-protein
proteins	NN	O	I-protein
correlates	NN	O	O
with	NN	O	O
hypo-acetylation	NN	O	O
of	NN	O	O
core	NN	O	B-protein
histones	NN	O	I-protein
at	NN	O	O
promoter	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
relieved	NN	O	O
by	NN	O	O
histone	NN	O	B-protein
deacetylase	NN	O	I-protein
inhibitors	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
Ikaros	NN	O	B-protein
and	NN	O	O
its	NN	O	O
repression	NN	O	B-protein
domains	NN	O	I-protein
can	NN	O	O
interact	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
the	NN	O	O
mSin3	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
co-repressors	NN	O	O
which	NN	O	O
bind	NN	O	O
to	NN	O	O
histone	NN	O	B-protein
deacetylases	NN	O	I-protein
.	NN	O	O

Based	NN	O	O
on	NN	O	O
these	NN	O	O
and	NN	O	O
our	NN	O	O
recent	NN	O	O
findings	NN	O	O
of	NN	O	O
associations	NN	O	O
between	NN	O	O
Ikaros	NN	O	B-protein
and	NN	O	O
Mi-2-HDAC	NN	O	B-protein
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
Ikaros	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
modulate	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
by	NN	O	O
recruiting	NN	O	O
distinct	NN	O	O
histone	NN	O	B-protein
deacetylase	NN	O	I-protein
complexes	NN	O	I-protein
to	NN	O	O
specific	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
c-Akt	NN	O	B-protein
)	NN	O	O
,	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
STAT5	NN	O	B-protein
are	NN	O	O
activated	NN	O	O
by	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
EPO	NN	O	B-protein
)	NN	O	O
in	NN	O	O
HCD57	NN	O	B-cell_line
erythroid	NN	O	I-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
are	NN	O	O
constitutively	NN	O	O
active	NN	O	O
in	NN	O	O
an	NN	O	O
EPO-independent	NN	O	B-cell_line
,	NN	O	I-cell_line
apoptosis-resistant	NN	O	I-cell_line
subclone	NN	O	I-cell_line
(	NN	O	O
HCD57-SREI	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
EPO	NN	O	B-protein
)	NN	O	O
and	NN	O	O
stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
SCF	NN	O	B-protein
)	NN	O	O
activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
PKB/Akt	NN	O	B-protein
)	NN	O	O
in	NN	O	O
EPO-dependent	NN	O	B-cell_line
HCD57	NN	O	I-cell_line
erythroid	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
better	NN	O	O
understand	NN	O	O
signals	NN	O	O
controlling	NN	O	O
proliferation	NN	O	O
and	NN	O	O
viability	NN	O	O
,	NN	O	O
erythroid	NN	O	O
cells	NN	O	O
that	NN	O	O
resist	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
EPO	NN	O	B-protein
were	NN	O	O
subcloned	NN	O	O
and	NN	O	O
characterized	NN	O	O
(	NN	O	O
HCD57-SREI	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
activations	NN	O	O
of	NN	O	O
PKB/Akt	NN	O	B-protein
,	NN	O	O
STAT5a	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT5b	NN	O	B-protein
were	NN	O	O
noted	NN	O	O
in	NN	O	O
these	NN	O	O
EPO-independent	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

PI3-kinase	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
an	NN	O	O
upstream	NN	O	O
activator	NN	O	O
of	NN	O	O
PKB/Akt	NN	O	B-protein
because	NN	O	O
the	NN	O	O
PI3-kinase	NN	O	B-protein
inhibitor	NN	O	O
LY294002	NN	O	O
blocked	NN	O	O
both	NN	O	O
constitutive	NN	O	O
PKB/Akt	NN	O	B-protein
and	NN	O	O
factor-dependent	NN	O	O
PKB/Akt	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

The	NN	O	O
LY294002	NN	O	O
study	NN	O	O
showed	NN	O	O
that	NN	O	O
proliferation	NN	O	O
and	NN	O	O
viability	NN	O	O
of	NN	O	O
both	NN	O	O
HCD57-SREI	NN	O	B-cell_line
and	NN	O	O
HCD57	NN	O	B-cell_line
cells	NN	O	I-cell_line
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
PKB/Akt	NN	O	B-protein
;	NN	O	O
however	NN	O	O
,	NN	O	O
PKB/Akt	NN	O	B-protein
activity	NN	O	O
alone	NN	O	O
did	NN	O	O
not	NN	O	O
protect	NN	O	O
these	NN	O	O
cells	NN	O	O
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
HCD57	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
SCF	NN	O	B-protein
also	NN	O	O
activated	NN	O	O
PKB/Akt	NN	O	B-protein
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
protect	NN	O	O
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

This	NN	O	O
result	NN	O	O
suggested	NN	O	O
that	NN	O	O
PKB/PI3-kinase	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
necessary	NN	O	O
but	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
promote	NN	O	O
viability	NN	O	O
and/or	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
STAT5	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
activated	NN	O	O
through	NN	O	O
an	NN	O	O
unknown	NN	O	O
pathway	NN	O	O
not	NN	O	O
including	NN	O	O
JAK2	NN	O	B-protein
or	NN	O	O
EPOR	NN	O	B-protein
,	NN	O	O
may	NN	O	O
act	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
the	NN	O	O
constitutive	NN	O	O
PI3-kinase	NN	O	B-protein
/PKB/Akt	NN	O	B-protein
pathway	NN	O	O
to	NN	O	O
protect	NN	O	O
the	NN	O	O
EPO-independent	NN	O	B-cell_line
HCD57-SREI	NN	O	I-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
promote	NN	O	O
limited	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Disruption	NN	O	O
of	NN	O	O
alpha	NN	O	O
beta	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
gamma	NN	O	O
delta	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
helix-loop-helix	NN	O	B-protein
protein	NN	O	I-protein
Id3	NN	O	B-protein
in	NN	O	O
committed	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

Enforced	NN	O	O
expression	NN	O	O
of	NN	O	O
Id3	NN	O	B-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
the	NN	O	O
capacity	NN	O	O
to	NN	O	O
inhibit	NN	O	O
many	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
(	NN	O	I-protein
bHLH	NN	O	I-protein
)	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
CD34	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
have	NN	O	O
not	NN	O	O
undergone	NN	O	O
T	NN	O	B-DNA
cell	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
TCR	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
rearrangements	NN	O	I-DNA
inhibits	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
transduced	NN	O	B-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
TCRalpha	NN	O	B-cell_line
beta	NN	O	I-cell_line
and	NN	O	I-cell_line
gamma	NN	O	I-cell_line
delta	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
a	NN	O	O
fetal	NN	O	B-cell_line
thymic	NN	O	I-cell_line
organ	NN	O	I-cell_line
culture	NN	O	I-cell_line
(	NN	O	O
FTOC	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
document	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Id3	NN	O	B-protein
,	NN	O	O
in	NN	O	O
progenitors	NN	O	B-cell_type
that	NN	O	O
have	NN	O	O
initiated	NN	O	O
TCR	NN	O	B-DNA
gene	NN	O	I-DNA
rearrangements	NN	O	I-DNA
(	NN	O	O
pre-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
,	NN	O	O
inhibits	NN	O	O
development	NN	O	O
into	NN	O	O
TCRalpha	NN	O	B-cell_line
beta	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
into	NN	O	O
TCRgamma	NN	O	B-cell_line
delta	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
Id3	NN	O	B-protein
impedes	NN	O	O
expression	NN	O	O
of	NN	O	O
recombination	NN	O	O
activating	NN	O	O
genes	NN	O	O
and	NN	O	O
downregulates	NN	O	O
pre-Talpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
possible	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
Id3	NN	O	B-protein
overexpression	NN	O	O
can	NN	O	O
differentially	NN	O	O
affect	NN	O	O
development	NN	O	O
of	NN	O	O
pre-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
TCRalpha	NN	O	B-cell_line
beta	NN	O	I-cell_line
and	NN	O	I-cell_line
gamma	NN	O	I-cell_line
delta	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
also	NN	O	O
observed	NN	O	O
that	NN	O	O
cell	NN	O	B-cell_type
surface	NN	O	I-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
CD3	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
rearranged	NN	O	B-DNA
TCR	NN	O	I-DNA
genes	NN	O	I-DNA
developed	NN	O	O
from	NN	O	O
Id3-transduced	NN	O	B-cell_line
but	NN	O	O
not	NN	O	O
from	NN	O	O
control-transduced	NN	O	B-cell_line
pre-T	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
an	NN	O	O
FTOC	NN	O	B-cell_line
.	NN	O	O

These	NN	O	O
cells	NN	O	O
had	NN	O	O
properties	NN	O	O
of	NN	O	O
both	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	O
NK	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
pre-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
bHLH	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
required	NN	O	O
to	NN	O	O
control	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
after	NN	O	O
the	NN	O	O
T/NK	NN	O	O
developmental	NN	O	O
checkpoint	NN	O	O
.	NN	O	O

-DOCSTART-	O

HIV-1	NN	O	O
reactivation	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
infected	NN	O	O
adults	NN	O	O
upon	NN	O	O
in	NN	O	O
vitro	NN	O	O
CD4	NN	O	B-protein
cross-linking	NN	O	O
by	NN	O	O
ligands	NN	O	O
of	NN	O	O
the	NN	O	O
CDR2-loop	NN	O	B-protein
in	NN	O	O
extracellular	NN	O	B-protein
domain	NN	O	I-protein
1	NN	O	I-protein
.	NN	O	O

HIV-1	NN	O	O
infects	NN	O	O
resting	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
but	NN	O	O
remains	NN	O	O
inactive	NN	O	O
state	NN	O	O
until	NN	O	O
subsequent	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
CD4	NN	O	I-protein
by	NN	O	O
gp120-anti-gp120	NN	O	B-protein
immune	NN	O	I-protein
complexes	NN	O	I-protein
or	NN	O	O
heat-inactivated	NN	O	O
HIV-1	NN	O	O
(	NN	O	O
iHIV-1	NN	O	O
)	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
trigger	NN	O	O
activation	NN	O	O
signals	NN	O	O
leading	NN	O	O
to	NN	O	O
virus	NN	O	O
reactivation	NN	O	O
(	NN	O	O
9	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
stimulation	NN	O	O
of	NN	O	O
virus	NN	O	O
production	NN	O	O
by	NN	O	O
iHIV-1	NN	O	O
were	NN	O	O
strictly	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
concentrations	NN	O	O
of	NN	O	O
viral	NN	O	B-protein
proteins	NN	O	I-protein
used	NN	O	O
as	NN	O	O
exogenous	NN	O	O
stimuli	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
further	NN	O	O
investigated	NN	O	O
the	NN	O	O
physiologic	NN	O	O
relevance	NN	O	O
of	NN	O	O
these	NN	O	O
observations	NN	O	O
.	NN	O	O

When	NN	O	O
submitted	NN	O	O
to	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
CD4	NN	O	B-protein
cross-linking	NN	O	O
by	NN	O	O
iHIV-1	NN	O	O
,	NN	O	O
PBMCs	NN	O	B-cell_type
from	NN	O	O
HIV-1-infected	NN	O	O
patients	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
produce	NN	O	O
virus	NN	O	O
.	NN	O	O

This	NN	O	O
viral	NN	O	O
reactivation	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
increased	NN	O	O
NF-kappaB	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
in	NN	O	O
patients	NN	O	O
'	NN	O	O
PBMCs	NN	O	B-cell_type
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
virus	NN	O	O
reactivation	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
PBMCs	NN	O	I-cell_type
infected	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
specifically	NN	O	O
induced	NN	O	O
by	NN	O	O
ligands	NN	O	O
of	NN	O	O
the	NN	O	O
CDR2-loop	NN	O	B-protein
in	NN	O	O
domain	NN	O	B-protein
1	NN	O	I-protein
(	NN	O	O
D1	NN	O	B-protein
)	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
(	NN	O	O
virus	NN	O	B-protein
envelope	NN	O	I-protein
and	NN	O	I-protein
anti-CD4	NN	O	I-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
virus	NN	O	O
reactivation	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
following	NN	O	O
CD4	NN	O	B-protein
oligomerization	NN	O	O
by	NN	O	O
antibodies	NN	O	B-protein
that	NN	O	O
bind	NN	O	O
other	NN	O	O
epitopes	NN	O	O
in	NN	O	O
D1	NN	O	B-protein
,	NN	O	O
including	NN	O	O
the	NN	O	O
D1/CDR3-loop	NN	O	B-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
soluble	NN	O	B-protein
CD4	NN	O	I-protein
(	NN	O	O
sCD4	NN	O	B-protein
)	NN	O	O
prevented	NN	O	O
virus	NN	O	O
reactivation	NN	O	O
by	NN	O	O
D1/CDR2-loop	NN	O	B-protein
ligands	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
signaling	NN	O	O
events	NN	O	O
initiated	NN	O	O
in	NN	O	O
PBMCs	NN	O	B-cell_type
by	NN	O	O
oligomerization	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
at	NN	O	O
the	NN	O	O
D1/CDR2-loop	NN	O	B-protein
can	NN	O	O
trigger	NN	O	O
HIV-1	NN	O	O
upregulation	NN	O	O
in	NN	O	O
infected	NN	O	O
individuals	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signaling	NN	O	O
through	NN	O	O
the	NN	O	O
lymphotoxin-beta	NN	O	B-protein
receptor	NN	O	I-protein
stimulates	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
alone	NN	O	O
and	NN	O	O
in	NN	O	O
cooperation	NN	O	O
with	NN	O	O
soluble	NN	O	O
or	NN	O	O
membrane-bound	NN	O	B-protein
TNF-alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
level	NN	O	O
of	NN	O	O
ongoing	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
within	NN	O	O
an	NN	O	O
individual	NN	O	O
is	NN	O	O
critical	NN	O	O
to	NN	O	O
HIV-1	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

Among	NN	O	O
host	NN	O	O
immune	NN	O	O
factors	NN	O	O
,	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
TNF-alpha	NN	O	B-protein
has	NN	O	O
previously	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
increase	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
various	NN	O	O
monocyte	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
model	NN	O	O
systems	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
signaling	NN	O	O
through	NN	O	O
the	NN	O	O
TNF	NN	O	B-protein
receptor	NN	O	O
family	NN	O	O
member	NN	O	O
,	NN	O	O
the	NN	O	O
lymphotoxin-beta	NN	O	B-protein
(	NN	O	I-protein
LT-beta	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
LT-betaR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
also	NN	O	O
regulates	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
is	NN	O	O
cooperatively	NN	O	O
stimulated	NN	O	O
when	NN	O	O
the	NN	O	O
distinct	NN	O	O
LT-betaR	NN	O	B-protein
and	NN	O	O
TNF	NN	O	B-protein
receptor	NN	O	I-protein
systems	NN	O	I-protein
are	NN	O	O
simultaneously	NN	O	O
engaged	NN	O	O
by	NN	O	O
their	NN	O	O
specific	NN	O	O
ligands	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
physiological	NN	O	O
coculture	NN	O	O
cellular	NN	O	O
assay	NN	O	O
system	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
membrane-bound	NN	O	B-protein
TNF-alpha	NN	O	I-protein
and	NN	O	O
LT-alpha1beta2	NN	O	B-protein
act	NN	O	O
virtually	NN	O	O
identically	NN	O	O
to	NN	O	O
their	NN	O	O
soluble	NN	O	O
forms	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
cosignaling	NN	O	O
via	NN	O	O
the	NN	O	O
LT-beta	NN	O	B-protein
and	NN	O	I-protein
TNF-alpha	NN	O	I-protein
receptors	NN	O	I-protein
is	NN	O	O
probably	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
and	NN	O	O
the	NN	O	O
subsequent	NN	O	O
determination	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
viral	NN	O	O
burden	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Intriguingly	NN	O	O
,	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
LT-alpha1beta2	NN	O	B-protein
is	NN	O	O
up-regulated	NN	O	O
on	NN	O	O
a	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
acutely	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
positive	NN	O	O
feedback	NN	O	O
loop	NN	O	O
between	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
,	NN	O	O
LT-alpha1beta2	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

Given	NN	O	O
the	NN	O	O
critical	NN	O	O
role	NN	O	O
that	NN	O	O
LT-alpha1beta2	NN	O	B-protein
plays	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
architecture	NN	O	O
,	NN	O	O
we	NN	O	O
speculate	NN	O	O
that	NN	O	O
LT-alpha1beta2	NN	O	B-protein
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
HIV-associated	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
the	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
.	NN	O	O

-DOCSTART-	O

Monoclonal	NN	O	B-protein
anti-endothelial	NN	O	I-protein
cell	NN	O	I-protein
antibodies	NN	O	I-protein
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
Takayasu	NN	O	O
arteritis	NN	O	O
activate	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
from	NN	O	O
large	NN	O	O
vessels	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
To	NN	O	O
create	NN	O	O
monoclonal	NN	O	B-protein
anti-endothelial	NN	O	I-protein
cell	NN	O	I-protein
antibodies	NN	O	I-protein
(	NN	O	O
mAECA	NN	O	B-protein
)	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
Takayasu	NN	O	O
arteritis	NN	O	O
to	NN	O	O
evaluate	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
activate	NN	O	O
human	NN	O	B-cell_line
umbilical	NN	O	I-cell_line
vein	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
HUVEC	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
EC	NN	O	O
activation	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
A	NN	O	O
panel	NN	O	O
of	NN	O	O
mAECA	NN	O	B-protein
was	NN	O	O
generated	NN	O	O
from	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
Takayasu	NN	O	O
arteritis	NN	O	O
,	NN	O	O
using	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
transformation	NN	O	O
.	NN	O	O

Activity	NN	O	O
against	NN	O	O
macrovascular	NN	O	B-cell_line
EC	NN	O	I-cell_line
(	NN	O	O
HUVEC	NN	O	B-cell_line
)	NN	O	O
and	NN	O	O
microvascular	NN	O	B-cell_line
EC	NN	O	I-cell_line
(	NN	O	O
human	NN	O	B-cell_line
bone	NN	O	I-cell_line
marrow	NN	O	I-cell_line
EC	NN	O	I-cell_line
immortalized	NN	O	I-cell_line
by	NN	O	I-cell_line
SV40	NN	O	I-cell_line
)	NN	O	O
antigens	NN	O	O
was	NN	O	O
detected	NN	O	O
by	NN	O	O
enzyme-linked	NN	O	O
immunosorbent	NN	O	O
assay	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
studies	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
select	NN	O	O
the	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
(	NN	O	O
mAECA	NN	O	B-protein
)	NN	O	O
which	NN	O	O
share	NN	O	O
the	NN	O	O
same	NN	O	O
EC	NN	O	O
epitope	NN	O	O
binding	NN	O	O
specificity	NN	O	O
as	NN	O	O
the	NN	O	O
total	NN	O	B-protein
IgG-AECA	NN	O	I-protein
from	NN	O	O
the	NN	O	O
Takayasu	NN	O	O
arteritis	NN	O	O
patient	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
mAECA	NN	O	B-protein
to	NN	O	O
human	NN	O	B-cell_type
aortic	NN	O	I-cell_type
EC	NN	O	I-cell_type
was	NN	O	O
studied	NN	O	O
by	NN	O	O
immunohistochemistry	NN	O	O
.	NN	O	O

The	NN	O	O
secretion	NN	O	O
levels	NN	O	O
of	NN	O	O
interleukin-6	NN	O	B-protein
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
and	NN	O	O
von	NN	O	O
Willebrand	NN	O	O
factor	NN	O	O
(	NN	O	O
vWF	NN	O	O
)	NN	O	O
were	NN	O	O
determined	NN	O	O
,	NN	O	O
to	NN	O	O
serve	NN	O	O
as	NN	O	O
markers	NN	O	O
for	NN	O	O
EC	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
activated	NN	O	B-cell_line
EC	NN	O	I-cell_line
were	NN	O	O
examined	NN	O	O
for	NN	O	O
the	NN	O	O
adherence	NN	O	O
of	NN	O	O
a	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
U937	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
intercellular	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
1	NN	O	O
,	NN	O	O
and	NN	O	O
E-selectin	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
the	NN	O	O
mAECA-treated	NN	O	B-cell_line
EC	NN	O	I-cell_line
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
translocation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
specific	NN	O	O
NF-kappaB	NN	O	B-protein
oligoprobe	NN	O	O
in	NN	O	O
an	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Six	NN	O	O
mAECA	NN	O	B-protein
were	NN	O	O
selected	NN	O	O
,	NN	O	O
the	NN	O	O
mixture	NN	O	O
of	NN	O	O
which	NN	O	O
produced	NN	O	O
100	NN	O	O
%	NN	O	O
inhibition	NN	O	O
of	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
original	NN	O	O
IgG	NN	O	B-protein
(	NN	O	O
from	NN	O	O
the	NN	O	O
patient	NN	O	O
with	NN	O	O
Takayasu	NN	O	O
arteritis	NN	O	O
)	NN	O	O
to	NN	O	O
HUVEC	NN	O	B-cell_line
.	NN	O	O

All	NN	O	O
mAECA	NN	O	B-protein
possessed	NN	O	O
high	NN	O	O
activity	NN	O	O
against	NN	O	O
macrovascular	NN	O	B-cell_line
EC	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
none	NN	O	O
had	NN	O	O
significant	NN	O	O
antimicrovascular	NN	O	O
EC	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
mAECA	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
normal	NN	O	B-protein
human	NN	O	I-protein
IgG	NN	O	I-protein
,	NN	O	O
had	NN	O	O
anti-human	NN	O	O
aortic	NN	O	O
EC	NN	O	O
activity	NN	O	O
.	NN	O	O

Four	NN	O	O
of	NN	O	O
the	NN	O	O
6	NN	O	O
mAECA	NN	O	B-protein
activated	NN	O	B-cell_line
EC	NN	O	I-cell_line
,	NN	O	O
manifested	NN	O	O
by	NN	O	O
increased	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
vWF	NN	O	O
secretion	NN	O	O
.	NN	O	O

The	NN	O	O
4	NN	O	O
mAECA	NN	O	B-protein
induced	NN	O	O
EC	NN	O	O
expression	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
and	NN	O	O
increased	NN	O	O
adhesion	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
EC	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
these	NN	O	O
mAECA	NN	O	B-protein
stimulated	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
AECA	NN	O	B-protein
may	NN	O	O
directly	NN	O	O
stimulate	NN	O	O
EC	NN	O	B-cell_type
in	NN	O	O
Takayasu	NN	O	O
arteritis	NN	O	O
through	NN	O	O
elevation	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
expression	NN	O	O
associated	NN	O	O
with	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
adhesion	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
may	NN	O	O
therefore	NN	O	O
play	NN	O	O
a	NN	O	O
pathogenic	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
vasculopathy	NN	O	O
in	NN	O	O
Takayasu	NN	O	O
arteritis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cutting	NN	O	O
edge	NN	O	O
:	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
in	NN	O	O
eosinophils	NN	O	B-cell_type
:	NN	O	O
regulation	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-5	NN	O	B-protein
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
NF-AT1	NN	O	B-protein
and	NN	O	O
NF-AT4	NN	O	B-protein
are	NN	O	O
expressed	NN	O	O
cytoplasmically	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
NF-AT2	NN	O	B-protein
and	NN	O	O
NF-AT3	NN	O	B-protein
have	NN	O	O
not	NN	O	O
been	NN	O	O
seen	NN	O	O
.	NN	O	O

Likewise	NN	O	O
,	NN	O	O
NF-AT1	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
NF-AT4	NN	O	B-RNA
mRNA	NN	O	I-RNA
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
their	NN	O	O
levels	NN	O	O
can	NN	O	O
be	NN	O	O
significantly	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
the	NN	O	O
Th2-associated	NN	O	B-protein
cytokines	NN	O	I-protein
IL-4	NN	O	B-protein
and	NN	O	O
IL-5	NN	O	B-protein
.	NN	O	O

There	NN	O	O
is	NN	O	O
no	NN	O	O
detectable	NN	O	O
NF-AT	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
NF-ATs	NN	O	B-protein
appear	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
IL-4-	NN	O	B-cell_line
,	NN	O	I-cell_line
IL-5-	NN	O	I-cell_line
,	NN	O	I-cell_line
or	NN	O	I-cell_line
ionomycin-stimulated	NN	O	I-cell_line
eosinophils	NN	O	I-cell_line
.	NN	O	O

Only	NN	O	O
NF-AT1	NN	O	B-protein
and	NN	O	O
NF-AT4	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
NF-AT2	NN	O	B-protein
and	NN	O	O
NF-AT3	NN	O	B-protein
,	NN	O	O
have	NN	O	O
translocated	NN	O	O
into	NN	O	O
the	NN	O	O
nuclei	NN	O	O
in	NN	O	O
IL-4-	NN	O	B-cell_line
or	NN	O	I-cell_line
IL-5-stimulated	NN	O	I-cell_line
eosinophils	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
findings	NN	O	O
delineate	NN	O	O
a	NN	O	O
novel	NN	O	O
pathway	NN	O	O
in	NN	O	O
the	NN	O	O
cytokine	NN	O	O
network	NN	O	O
in	NN	O	O
which	NN	O	O
Th2	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
``	NN	O	O
control	NN	O	O
''	NN	O	O
eosinophils	NN	O	B-cell_type
via	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-5	NN	O	B-protein
,	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
in	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
findings	NN	O	O
also	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
later	NN	O	O
feedback	NN	O	O
``	NN	O	O
talking	NN	O	O
''	NN	O	O
may	NN	O	O
exist	NN	O	O
between	NN	O	O
eosinophils	NN	O	B-cell_type
and	NN	O	O
Th2	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

PPARalpha	NN	O	O
activators	NN	O	O
inhibit	NN	O	O
cytokine-induced	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Adhesion	NN	O	O
molecule	NN	O	O
expression	NN	O	O
on	NN	O	O
the	NN	O	O
endothelial	NN	O	O
cell	NN	O	O
(	NN	O	O
EC	NN	O	O
)	NN	O	O
surface	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
leukocyte	NN	O	O
recruitment	NN	O	O
to	NN	O	O
atherosclerotic	NN	O	O
lesions	NN	O	O
.	NN	O	O

Better	NN	O	O
understanding	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
in	NN	O	O
ECs	NN	O	B-cell_type
may	NN	O	O
provide	NN	O	O
important	NN	O	O
insight	NN	O	O
into	NN	O	O
plaque	NN	O	O
formation	NN	O	O
.	NN	O	O

Peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor-alpha	NN	O	I-protein
(	NN	O	O
PPARalpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
regulates	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
certain	NN	O	O
fatty	NN	O	O
acids	NN	O	O
and	NN	O	O
fibric	NN	O	O
acid	NN	O	O
derivatives	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
investigated	NN	O	O
PPARalpha	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
ECs	NN	O	I-cell_type
and	NN	O	O
their	NN	O	O
regulation	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

METHODS	NN	O	O
AND	NN	O	O
RESULTS	NN	O	O
:	NN	O	O
Immunohistochemistry	NN	O	O
revealed	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
carotid	NN	O	I-cell_type
artery	NN	O	I-cell_type
ECs	NN	O	I-cell_type
express	NN	O	O
PPARalpha	NN	O	B-protein
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
cultured	NN	O	B-cell_type
human	NN	O	I-cell_type
ECs	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
PPARalpha	NN	O	B-protein
activators	NN	O	O
fenofibrate	NN	O	O
or	NN	O	O
WY14643	NN	O	O
inhibited	NN	O	O
TNF-alpha-induced	NN	O	B-protein
VCAM-1	NN	O	I-protein
in	NN	O	O
a	NN	O	O
time-	NN	O	O
and	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
an	NN	O	O
effect	NN	O	O
not	NN	O	O
seen	NN	O	O
with	NN	O	O
PPARgamma	NN	O	B-protein
activators	NN	O	O
.	NN	O	O

Both	NN	O	O
PPARalpha	NN	O	B-protein
activators	NN	O	O
decreased	NN	O	O
cytokine-induced	NN	O	O
VCAM-1	NN	O	B-protein
mRNA	NN	O	O
expression	NN	O	O
without	NN	O	O
altering	NN	O	O
its	NN	O	O
mRNA	NN	O	O
half-life	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
deletional	NN	O	O
VCAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
suggest	NN	O	O
that	NN	O	O
fenofibrate	NN	O	O
inhibits	NN	O	O
VCAM-1	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
PPARalpha	NN	O	B-protein
activators	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
adhesion	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
cultured	NN	O	B-cell_type
human	NN	O	I-cell_type
ECs	NN	O	I-cell_type
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Human	NN	O	B-cell_type
ECs	NN	O	I-cell_type
express	NN	O	O
PPARalpha	NN	O	B-protein
,	NN	O	O
a	NN	O	O
potentially	NN	O	O
important	NN	O	O
regulator	NN	O	O
of	NN	O	O
atherogenesis	NN	O	O
through	NN	O	O
its	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Such	NN	O	O
findings	NN	O	O
also	NN	O	O
have	NN	O	O
implications	NN	O	O
regarding	NN	O	O
the	NN	O	O
clinical	NN	O	O
use	NN	O	O
of	NN	O	O
lipid-lowering	NN	O	O
agents	NN	O	O
,	NN	O	O
like	NN	O	O
fibric	NN	O	O
acids	NN	O	O
,	NN	O	O
which	NN	O	O
can	NN	O	O
activate	NN	O	O
PPARalpha	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

CBP/p300	NN	O	B-protein
integrates	NN	O	O
Raf	NN	O	B-protein
/	NN	O	O
Rac	NN	O	B-protein
-signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-ATc	NN	O	B-protein
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

NF-ATc	NN	O	B-protein
,	NN	O	O
an	NN	O	O
inducibly	NN	O	O
expressed	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
controls	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
cardiomyocytes	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
co-activators	NN	O	I-protein
CBP/p300	NN	O	B-protein
bind	NN	O	O
to	NN	O	O
and	NN	O	O
control	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	B-protein
N-terminal	NN	O	I-protein
transactivation	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
NF-ATc	NN	O	B-protein
,	NN	O	O
TAD-A	NN	O	B-protein
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
the	NN	O	O
N	NN	O	B-protein
terminal	NN	O	I-protein
transactivation	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
TAD-A	NN	O	B-protein
is	NN	O	O
inducibly	NN	O	O
phosphorylated	NN	O	O
,	NN	O	O
but	NN	O	O
this	NN	O	O
phosphorylation	NN	O	O
is	NN	O	O
dispensable	NN	O	O
for	NN	O	O
the	NN	O	O
interaction	NN	O	O
with	NN	O	O
CBP/p300	NN	O	B-protein
.	NN	O	O

Constitutive	NN	O	O
active	NN	O	O
versions	NN	O	O
of	NN	O	O
c-Raf	NN	O	B-protein
and	NN	O	O
Rac	NN	O	B-protein
synergistically	NN	O	O
enhance	NN	O	O
the	NN	O	O
CBP/p300	NN	O	B-protein
-mediated	NN	O	O
increase	NN	O	O
of	NN	O	O
TAD-A	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
important	NN	O	O
role	NN	O	O
CBP/p300	NN	O	B-protein
plays	NN	O	O
in	NN	O	O
the	NN	O	O
integration	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
.	NN	O	O

Since	NN	O	O
a	NN	O	O
mutation	NN	O	O
of	NN	O	O
CBP	NN	O	B-protein
abolishing	NN	O	O
HAT	NN	O	O
activity	NN	O	O
is	NN	O	O
almost	NN	O	O
as	NN	O	O
active	NN	O	O
as	NN	O	O
wild-type	NN	O	B-protein
CBP	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
functions	NN	O	O
of	NN	O	O
CBP/p300	NN	O	B-protein
other	NN	O	O
than	NN	O	O
histone	NN	O	O
acetylation	NN	O	O
appear	NN	O	O
to	NN	O	O
control	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Immune	NN	O	O
functions	NN	O	O
,	NN	O	O
clinical	NN	O	O
parameters	NN	O	O
and	NN	O	O
hormone	NN	O	O
receptor	NN	O	O
status	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
carried	NN	O	O
out	NN	O	O
a	NN	O	O
detailed	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	O
immune	NN	O	O
functions	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

A	NN	O	O
possible	NN	O	O
correlation	NN	O	O
between	NN	O	O
immune	NN	O	O
and	NN	O	O
clinical	NN	O	O
parameters	NN	O	O
was	NN	O	O
analysed	NN	O	O
in	NN	O	O
50	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
.	NN	O	O

Immune	NN	O	O
parameters	NN	O	O
,	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cell	NN	O	I-cell_type
and	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
functions	NN	O	O
and	NN	O	O
the	NN	O	O
numbers	NN	O	O
of	NN	O	O
circulating	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
analysed	NN	O	O
against	NN	O	O
the	NN	O	O
clinical	NN	O	O
parameters	NN	O	O
comprising	NN	O	O
the	NN	O	O
tumour	NN	O	O
burden	NN	O	O
,	NN	O	O
the	NN	O	O
stage	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
hormone	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
the	NN	O	O
tumour	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
analyse	NN	O	O
the	NN	O	O
immune	NN	O	O
function	NN	O	O
data	NN	O	O
effectively	NN	O	O
,	NN	O	O
low	NN	O	O
responders	NN	O	O
were	NN	O	O
identified	NN	O	O
with	NN	O	O
stringent	NN	O	O
cut-off	NN	O	O
values	NN	O	O
.	NN	O	O

Considerably	NN	O	O
higher	NN	O	O
proportions	NN	O	O
of	NN	O	O
low	NN	O	O
responders	NN	O	O
were	NN	O	O
found	NN	O	O
among	NN	O	O
the	NN	O	O
patient	NN	O	O
population	NN	O	O
.	NN	O	O

Elevated	NN	O	O
numbers	NN	O	O
of	NN	O	O
circulating	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
CD3	NN	O	B-protein
-directed	NN	O	O
cytolysis	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
oestrogen	NN	O	B-protein
receptors	NN	O	I-protein
independently	NN	O	O
of	NN	O	O
the	NN	O	O
clinical/histological	NN	O	O
parameters	NN	O	O
.	NN	O	O

-DOCSTART-	O

Latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
interacts	NN	O	O
with	NN	O	O
JAK3	NN	O	B-protein
and	NN	O	O
activates	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
LMP1	NN	O	B-protein
)	NN	O	O
acts	NN	O	O
like	NN	O	O
a	NN	O	O
permanently	NN	O	B-protein
activated	NN	O	I-protein
receptor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
-receptor	NN	O	I-protein
superfamily	NN	O	I-protein
and	NN	O	O
is	NN	O	O
absolutely	NN	O	O
required	NN	O	O
for	NN	O	O
B	NN	O	O
cell	NN	O	O
immortalization	NN	O	O
by	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
.	NN	O	O

Molecular	NN	O	O
and	NN	O	O
biochemical	NN	O	O
approaches	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
LMP1	NN	O	B-protein
usurps	NN	O	O
cellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
via	NN	O	O
two	NN	O	O
C-terminal	NN	O	B-protein
activating	NN	O	I-protein
regions	NN	O	I-protein
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
that	NN	O	O
a	NN	O	O
third	NN	O	O
region	NN	O	O
encompassing	NN	O	O
a	NN	O	O
proline	NN	O	B-protein
rich	NN	O	I-protein
sequence	NN	O	I-protein
within	NN	O	O
the	NN	O	O
33	NN	O	O
bp	NN	O	O
repetitive	NN	O	O
stretch	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
's	NN	O	O
C-terminus	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
JAK3	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
interaction	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
and	NN	O	O
JAK3	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
the	NN	O	O
enhanced	NN	O	O
tyrosine	NN	O	O
auto/transphosphorylation	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
within	NN	O	O
minutes	NN	O	O
after	NN	O	O
crosslinking	NN	O	O
of	NN	O	O
a	NN	O	O
conditional	NN	O	O
NGF-R	NN	O	B-protein
:	NN	O	I-protein
LMP1	NN	O	I-protein
chimera	NN	O	I-protein
and	NN	O	O
is	NN	O	O
a	NN	O	O
prerequisite	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
reveal	NN	O	O
a	NN	O	O
novel	NN	O	O
activating	NN	O	O
region	NN	O	O
in	NN	O	O
the	NN	O	O
LMP1	NN	O	B-protein
C-terminus	NN	O	B-protein
and	NN	O	O
identify	NN	O	O
the	NN	O	O
JAK	NN	O	B-protein
/STAT	NN	O	B-protein
pathway	NN	O	O
as	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
this	NN	O	O
viral	NN	O	B-protein
integral	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Monocyte	NN	O	O
arrest	NN	O	O
and	NN	O	O
transmigration	NN	O	O
on	NN	O	O
inflamed	NN	O	O
endothelium	NN	O	O
in	NN	O	O
shear	NN	O	O
flow	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
adenovirus-mediated	NN	O	O
gene	NN	O	O
transfer	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
.	NN	O	O

Mobilization	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
activates	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
encoding	NN	O	O
endothelial	NN	O	B-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
and	NN	O	O
chemokines	NN	O	O
that	NN	O	O
contribute	NN	O	O
to	NN	O	O
monocyte	NN	O	O
infiltration	NN	O	O
critical	NN	O	O
in	NN	O	O
atherogenesis	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
has	NN	O	O
been	NN	O	O
achieved	NN	O	O
by	NN	O	O
pharmacological	NN	O	O
and	NN	O	O
genetic	NN	O	O
approaches	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
monocyte	NN	O	O
interactions	NN	O	O
with	NN	O	O
activated	NN	O	O
endothelium	NN	O	O
in	NN	O	O
shear	NN	O	O
flow	NN	O	O
following	NN	O	O
gene	NN	O	O
transfer	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
inhibitor	NN	O	O
IkappaB-alpha	NN	O	B-protein
have	NN	O	O
not	NN	O	O
been	NN	O	O
studied	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
a	NN	O	O
recombinant	NN	O	O
adenovirus	NN	O	O
prevented	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
-induced	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
and	NN	O	O
suppressed	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
E-selectin	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
surface	NN	O	O
protein	NN	O	O
expression	NN	O	O
and	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
transcripts	NN	O	O
for	NN	O	O
the	NN	O	O
chemokines	NN	O	B-protein
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
MCP-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
growth-related	NN	O	B-protein
activity-alpha	NN	O	I-protein
(	NN	O	O
GRO-alpha	NN	O	B-protein
)	NN	O	O
by	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

This	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
reduction	NN	O	O
in	NN	O	O
endothelial	NN	O	O
MCP-1	NN	O	B-protein
secretion	NN	O	O
and	NN	O	O
GRO-alpha	NN	O	B-protein
immobilization	NN	O	O
.	NN	O	O

Adhesion	NN	O	O
assays	NN	O	O
under	NN	O	O
physiological	NN	O	O
shear	NN	O	O
flow	NN	O	O
conditions	NN	O	O
showed	NN	O	O
that	NN	O	O
firm	NN	O	O
arrest	NN	O	O
,	NN	O	O
spreading	NN	O	O
,	NN	O	O
and	NN	O	O
transmigration	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
on	NN	O	O
TNF-alpha	NN	O	B-protein
-activated	NN	O	O
endothelium	NN	O	O
was	NN	O	O
markedly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
IkappaB-alpha	NN	O	B-protein
overexpression	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
with	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
and	NN	O	O
peptide	NN	O	O
antagonists	NN	O	O
inferred	NN	O	O
that	NN	O	O
this	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
reduced	NN	O	O
expression	NN	O	O
of	NN	O	O
Ig	NN	O	O
integrin	NN	O	O
ligand	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
of	NN	O	O
chemokines	NN	O	B-protein
specifically	NN	O	O
involved	NN	O	O
in	NN	O	O
these	NN	O	O
events	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
rolling	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
was	NN	O	O
increased	NN	O	O
by	NN	O	O
IkappaB-alpha	NN	O	B-protein
transfer	NN	O	O
and	NN	O	O
was	NN	O	O
partly	NN	O	O
mediated	NN	O	O
by	NN	O	O
P-selectin	NN	O	B-protein
;	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
unaffected	NN	O	O
by	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
provide	NN	O	O
novel	NN	O	O
evidence	NN	O	O
that	NN	O	O
selective	NN	O	O
modulation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
by	NN	O	O
adenoviral	NN	O	O
transfer	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
impairs	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
multiple	NN	O	O
endothelial	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
required	NN	O	O
for	NN	O	O
subsequent	NN	O	O
monocyte	NN	O	O
arrest	NN	O	O
and	NN	O	O
emigration	NN	O	O
in	NN	O	O
shear	NN	O	O
flow	NN	O	O
and	NN	O	O
thus	NN	O	O
for	NN	O	O
monocyte	NN	O	O
infiltration	NN	O	O
in	NN	O	O
atherosclerotic	NN	O	O
plaques	NN	O	O
.	NN	O	O

-DOCSTART-	O

Recognition	NN	O	O
of	NN	O	O
NFATp/AP-1	NN	O	B-DNA
composite	NN	O	I-DNA
elements	NN	O	I-DNA
within	NN	O	O
genes	NN	O	O
induced	NN	O	O
upon	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Composite	NN	O	B-DNA
elements	NN	O	I-DNA
are	NN	O	O
regulatory	NN	O	B-DNA
modules	NN	O	I-DNA
of	NN	O	O
promoters	NN	O	B-DNA
or	NN	O	O
enhancers	NN	O	B-DNA
that	NN	O	O
consist	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
of	NN	O	O
two	NN	O	O
different	NN	O	O
but	NN	O	O
synergizing	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

A	NN	O	O
well-studied	NN	O	O
example	NN	O	O
is	NN	O	O
nuclear	NN	O	B-DNA
factors	NN	O	I-DNA
of	NN	O	I-DNA
activated	NN	O	I-DNA
T-cell	NN	O	I-DNA
(	NN	O	I-DNA
NFAT	NN	O	I-DNA
)	NN	O	I-DNA
sites	NN	O	I-DNA
which	NN	O	O
are	NN	O	O
composite	NN	O	O
elements	NN	O	O
of	NN	O	O
a	NN	O	O
NFATp/c	NN	O	B-DNA
and	NN	O	I-DNA
an	NN	O	I-DNA
activating	NN	O	I-DNA
protein	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	I-DNA
AP-1	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
computational	NN	O	O
approach	NN	O	O
to	NN	O	O
identify	NN	O	O
potential	NN	O	O
NFAT	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
which	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
comprises	NN	O	O
an	NN	O	O
improved	NN	O	O
method	NN	O	O
to	NN	O	O
scan	NN	O	O
for	NN	O	O
individual	NN	O	O
NFAT	NN	O	B-DNA
composite	NN	O	I-DNA
elements	NN	O	I-DNA
;	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
considers	NN	O	O
positional	NN	O	O
effects	NN	O	O
relative	NN	O	O
to	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
sites	NN	O	I-DNA
;	NN	O	O
and	NN	O	O
(	NN	O	O
c	NN	O	O
)	NN	O	O
involves	NN	O	O
cluster	NN	O	O
analysis	NN	O	O
of	NN	O	O
potential	NN	O	O
NFAT	NN	O	B-DNA
composite	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

All	NN	O	O
three	NN	O	O
steps	NN	O	O
progressively	NN	O	O
helpX	NN	O	O
?	NN	O	O
ed	NN	O	O
to	NN	O	O
discriminate	NN	O	O
T-cell-specific	NN	O	B-DNA
promoter	NN	O	I-DNA
sequences	NN	O	I-DNA
against	NN	O	O
other	NN	O	O
functional	NN	O	B-DNA
regions	NN	O	I-DNA
(	NN	O	O
coding	NN	O	B-DNA
and	NN	O	I-DNA
intronic	NN	O	I-DNA
sequences	NN	O	I-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
genes	NN	O	O
,	NN	O	O
against	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
muscle-specific	NN	O	B-DNA
genes	NN	O	I-DNA
or	NN	O	O
against	NN	O	O
random	NN	O	B-DNA
sequences	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
this	NN	O	O
approach	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
potential	NN	O	O
NFAT	NN	O	B-DNA
composite	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
their	NN	O	O
receptors	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
genes	NN	O	O
for	NN	O	O
AP-1	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
,	NN	O	O
Ca2+-binding	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
some	NN	O	O
other	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
regulatory	NN	O	O
network	NN	O	O
operating	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
and	NN	O	O
other	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
method	NN	O	O
developed	NN	O	O
can	NN	O	O
be	NN	O	O
adapted	NN	O	O
to	NN	O	O
characterize	NN	O	O
and	NN	O	O
identify	NN	O	O
other	NN	O	O
composite	NN	O	B-DNA
elements	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
.	NN	O	O

The	NN	O	O
program	NN	O	O
for	NN	O	O
recognition	NN	O	O
NFAT	NN	O	B-DNA
composite	NN	O	I-DNA
elements	NN	O	I-DNA
is	NN	O	O
available	NN	O	O
through	NN	O	O
the	NN	O	O
World	NN	O	O
Wide	NN	O	O
Web	NN	O	O
(	NN	O	O
http	NN	O	O
:	NN	O	O
//compel.bionet.nsc.ru/FunSite/CompelScan.html	NN	O	O
and	NN	O	O
http	NN	O	O
:	NN	O	O
//transfac.gbf.de/dbsearch/funsitep/s_comp.html	NN	O	O
)	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
oestrogen	NN	O	B-protein
receptor	NN	O	I-protein
codon	NN	O	B-DNA
10	NN	O	I-DNA
polymorphism	NN	O	O
detected	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
is	NN	O	O
also	NN	O	O
present	NN	O	O
in	NN	O	O
non-malignant	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
oestrogens	NN	O	O
on	NN	O	O
oestrogen-receptive	NN	O	O
organs	NN	O	O
and	NN	O	O
cells	NN	O	O
is	NN	O	O
mediated	NN	O	O
via	NN	O	O
intracellular	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
ERalpha	NN	O	B-protein
and	NN	O	O
ERbeta	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Oestrogen	NN	O	B-protein
receptor	NN	O	I-protein
gene	NN	O	O
polymorphisms	NN	O	O
in	NN	O	O
the	NN	O	O
region	NN	O	O
encoding	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
portion	NN	O	I-protein
of	NN	O	O
the	NN	O	O
protein	NN	O	O
are	NN	O	O
reportedly	NN	O	O
associated	NN	O	O
with	NN	O	O
pathological	NN	O	O
conditions	NN	O	O
including	NN	O	O
breast	NN	O	O
cancer	NN	O	O
,	NN	O	O
hypertension	NN	O	O
,	NN	O	O
spontaneous	NN	O	O
abortion	NN	O	O
and	NN	O	O
coronary	NN	O	O
heart	NN	O	O
disease	NN	O	O
.	NN	O	O

A	NN	O	O
silent	NN	O	O
mutation	NN	O	O
in	NN	O	O
codon	NN	O	B-DNA
10	NN	O	I-DNA
of	NN	O	O
exon	NN	O	B-DNA
1	NN	O	I-DNA
,	NN	O	O
detected	NN	O	O
in	NN	O	O
ER-negative	NN	O	B-cell_line
and	NN	O	I-cell_line
ER-positive	NN	O	I-cell_line
human	NN	O	I-cell_line
breast	NN	O	I-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
breast	NN	O	O
tumors	NN	O	O
and	NN	O	O
blood	NN	O	B-DNA
DNA	NN	O	I-DNA
from	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
recognized	NN	O	O
as	NN	O	O
a	NN	O	O
polymorphic	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
examined	NN	O	O
,	NN	O	O
by	NN	O	O
denaturing	NN	O	O
gradient-gel	NN	O	O
electrophoresis	NN	O	O
and	NN	O	O
DNA	NN	O	B-DNA
sequence	NN	O	I-DNA
analysis	NN	O	O
,	NN	O	O
the	NN	O	O
possible	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
codon	NN	O	B-DNA
10	NN	O	I-DNA
polymorphic	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
normal	NN	O	O
oestrogen	NN	O	O
target	NN	O	O
organs	NN	O	O
and	NN	O	O
cells	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
uterus	NN	O	O
(	NN	O	O
myometrium	NN	O	O
and	NN	O	O
endometrium	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
placenta	NN	O	O
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
a	NN	O	O
benign	NN	O	O
uterus	NN	O	O
tumour	NN	O	O
(	NN	O	O
leiomyoma	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
detected	NN	O	O
ER	NN	O	B-DNA
codon	NN	O	I-DNA
10	NN	O	I-DNA
polymorphism	NN	O	O
in	NN	O	O
these	NN	O	O
samples	NN	O	O
and	NN	O	O
have	NN	O	O
compared	NN	O	O
them	NN	O	O
to	NN	O	O
those	NN	O	O
observed	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
samples	NN	O	O
.	NN	O	O

All	NN	O	O
tissues	NN	O	O
and	NN	O	O
cells	NN	O	O
studied	NN	O	O
were	NN	O	O
homozygous	NN	O	O
for	NN	O	O
the	NN	O	O
wild-type	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
homozygous	NN	O	O
for	NN	O	O
the	NN	O	O
codon-10-variant	NN	O	B-DNA
type	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
codon-10-variant	NN	O	B-DNA
type	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
a	NN	O	O
characteristic	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

Out	NN	O	O
current	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
further	NN	O	O
investigations	NN	O	O
are	NN	O	O
warranted	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
possible	NN	O	O
linkage	NN	O	O
of	NN	O	O
ER	NN	O	B-DNA
codon	NN	O	I-DNA
10	NN	O	I-DNA
polymorphism	NN	O	O
to	NN	O	O
physiological	NN	O	O
and	NN	O	O
pathological	NN	O	O
conditions	NN	O	O
.	NN	O	O

-DOCSTART-	O

USF/c-Myc	NN	O	B-protein
enhances	NN	O	O
,	NN	O	O
while	NN	O	O
Yin-Yang	NN	O	B-protein
1	NN	O	I-protein
suppresses	NN	O	O
,	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
of	NN	O	O
CXCR4	NN	O	B-protein
,	NN	O	O
a	NN	O	O
coreceptor	NN	O	B-protein
for	NN	O	O
HIV-1	NN	O	O
entry	NN	O	O
.	NN	O	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
USF1	NN	O	B-protein
and	NN	O	O
USF2	NN	O	B-protein
up-regulate	NN	O	O
gene	NN	O	O
expression	NN	O	O
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeats	NN	O	I-DNA
)	NN	O	O
via	NN	O	O
interaction	NN	O	O
with	NN	O	O
an	NN	O	O
E	NN	O	B-DNA
box	NN	O	I-DNA
on	NN	O	O
their	NN	O	O
target	NN	O	O
promoters	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
also	NN	O	O
a	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
c-Myc	NN	O	B-protein
.	NN	O	O

The	NN	O	O
c-Myc	NN	O	B-protein
oncoprotein	NN	O	I-protein
is	NN	O	O
important	NN	O	O
in	NN	O	O
control	NN	O	O
of	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
,	NN	O	O
while	NN	O	O
Yin-Yang	NN	O	B-protein
1	NN	O	I-protein
(	NN	O	O
YY1	NN	O	B-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
control	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
cellular	NN	O	B-DNA
and	NN	O	I-DNA
viral	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
two	NN	O	O
proteins	NN	O	O
physically	NN	O	O
interact	NN	O	O
with	NN	O	O
each	NN	O	O
other	NN	O	O
and	NN	O	O
mutually	NN	O	O
inhibit	NN	O	O
their	NN	O	O
respective	NN	O	O
biological	NN	O	O
functions	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
USF/c-Myc	NN	O	B-protein
up-regulates	NN	O	O
,	NN	O	O
while	NN	O	O
YY1	NN	O	B-protein
down-regulates	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
of	NN	O	O
CXCR4	NN	O	B-protein
,	NN	O	O
a	NN	O	O
coreceptor	NN	O	O
for	NN	O	O
T	NN	O	O
cell-tropic	NN	O	O
HIV-1	NN	O	O
entry	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
an	NN	O	O
E	NN	O	B-DNA
box	NN	O	I-DNA
around	NN	O	O
-260	NN	O	B-DNA
and	NN	O	O
a	NN	O	O
YY1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
around	NN	O	O
-300	NN	O	B-DNA
relative	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
E	NN	O	B-DNA
box	NN	O	I-DNA
abolished	NN	O	O
USF/c-Myc	NN	O	B-protein
-mediated	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
CXCR4	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
YY1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
associated	NN	O	O
with	NN	O	O
unresponsiveness	NN	O	O
to	NN	O	O
YY1	NN	O	B-protein
-mediated	NN	O	O
inhibition	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
USF/c-Myc	NN	O	B-protein
and	NN	O	O
YY1	NN	O	B-protein
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
HIV-1-replicative	NN	O	O
cycle	NN	O	O
,	NN	O	O
by	NN	O	O
modulating	NN	O	O
both	NN	O	O
the	NN	O	O
viral	NN	O	O
fusion/entry	NN	O	O
process	NN	O	O
and	NN	O	O
viral	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
supernatant	NN	O	O
fluid	NN	O	O
from	NN	O	O
human	NN	O	O
renal	NN	O	O
cell	NN	O	O
carcinomas	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
renal	NN	O	O
cell	NN	O	O
carcinoma	NN	O	O
(	NN	O	O
RCC	NN	O	O
)	NN	O	O
show	NN	O	O
reduced	NN	O	O
immune	NN	O	O
function	NN	O	O
and	NN	O	O
impaired	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

We	NN	O	O
determined	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
suppression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
RCC	NN	O	O
patient	NN	O	O
and	NN	O	O
determined	NN	O	O
whether	NN	O	O
supernatant	NN	O	O
fluid	NN	O	O
from	NN	O	O
RCC	NN	O	O
explants	NN	O	O
(	NN	O	O
RCC-S	NN	O	O
)	NN	O	O
induced	NN	O	O
the	NN	O	O
same	NN	O	O
phenotype	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
suppression	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
is	NN	O	O
observed	NN	O	O
in	NN	O	O
patient	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
pattern	NN	O	O
of	NN	O	O
kappaB	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
RCC	NN	O	O
patient	NN	O	O
was	NN	O	O
altered	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
seen	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
normal	NN	O	O
volunteers	NN	O	O
.	NN	O	O

In	NN	O	O
some	NN	O	O
patients	NN	O	O
,	NN	O	O
no	NN	O	O
activation	NN	O	O
of	NN	O	O
RelA/NFkappaB1	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
was	NN	O	O
detectable	NN	O	O
,	NN	O	O
while	NN	O	O
in	NN	O	O
others	NN	O	O
kappaB	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
was	NN	O	O
modestly	NN	O	O
induced	NN	O	O
but	NN	O	O
the	NN	O	O
duration	NN	O	O
was	NN	O	O
reduced	NN	O	O
.	NN	O	O

IkappaBalpha	NN	O	B-protein
was	NN	O	O
degraded	NN	O	O
normally	NN	O	O
following	NN	O	O
stimulation	NN	O	O
in	NN	O	O
both	NN	O	O
normal	NN	O	O
controls	NN	O	O
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
RCC	NN	O	O
patients	NN	O	O
.	NN	O	O

RCC-S	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
levels	NN	O	O
of	NN	O	O
RelA	NN	O	B-protein
and	NN	O	O
NF-kappaB1	NN	O	B-protein
but	NN	O	O
did	NN	O	O
suppress	NN	O	O
their	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
and	NN	O	O
inhibited	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
RelA/NF-kappaB1	NN	O	B-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
RCC-S	NN	O	O
can	NN	O	O
induce	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
the	NN	O	O
same	NN	O	O
phenotype	NN	O	O
of	NN	O	O
impaired	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
that	NN	O	O
is	NN	O	O
detected	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
RCC	NN	O	O
patient	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
appears	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
suppression	NN	O	O
by	NN	O	O
RCC-S	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
immunosuppression	NN	O	O
of	NN	O	O
host	NN	O	O
immunity	NN	O	O
.	NN	O	O

-DOCSTART-	O

STAT1	NN	O	B-protein
activation	NN	O	O
during	NN	O	O
monocyte	NN	O	O
to	NN	O	O
macrophage	NN	O	O
maturation	NN	O	O
:	NN	O	O
role	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

Human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
healthy	NN	O	O
donors	NN	O	O
show	NN	O	O
a	NN	O	O
time-dependent	NN	O	O
differentiation	NN	O	O
into	NN	O	O
macrophages	NN	O	B-cell_type
upon	NN	O	O
in	NN	O	O
vitro	NN	O	O
cultivation	NN	O	O
,	NN	O	O
closely	NN	O	O
mimicking	NN	O	O
their	NN	O	O
in	NN	O	O
vivo	NN	O	O
migration	NN	O	O
and	NN	O	O
maturation	NN	O	O
into	NN	O	O
extravascular	NN	O	O
tissues	NN	O	O
.	NN	O	O

The	NN	O	O
mediator	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
this	NN	O	O
maturation	NN	O	O
process	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
yet	NN	O	O
defined	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
and	NN	O	O
reported	NN	O	O
the	NN	O	O
specific	NN	O	O
,	NN	O	O
time-dependent	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
protein	NN	O	O
starting	NN	O	O
at	NN	O	O
day	NN	O	O
0/1	NN	O	O
of	NN	O	O
cultivation	NN	O	O
and	NN	O	O
maximally	NN	O	O
expressed	NN	O	O
at	NN	O	O
day	NN	O	O
5	NN	O	O
.	NN	O	O

STAT1	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
evident	NN	O	O
on	NN	O	O
the	NN	O	O
STAT	NN	O	B-DNA
binding	NN	O	I-DNA
sequences	NN	O	I-DNA
(	NN	O	O
SBE	NN	O	B-DNA
)	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
genes	NN	O	O
which	NN	O	O
are	NN	O	O
up-regulated	NN	O	O
during	NN	O	O
monocyte	NN	O	O
to	NN	O	O
macrophage	NN	O	O
maturation	NN	O	O
such	NN	O	O
as	NN	O	O
FcgammaRI	NN	O	B-DNA
and	NN	O	O
ICAM-1	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
factor	NN	O	I-DNA
IFN	NN	O	I-DNA
regulatory	NN	O	I-DNA
factor-1	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
to	NN	O	O
fibronectin	NN	O	O
or	NN	O	O
laminin	NN	O	O
was	NN	O	O
studied	NN	O	O
to	NN	O	O
investigate	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
STAT1	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Compared	NN	O	O
with	NN	O	O
monocytes	NN	O	B-cell_type
adherent	NN	O	O
on	NN	O	O
plastic	NN	O	O
surfaces	NN	O	O
,	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
cells	NN	O	O
allowed	NN	O	O
to	NN	O	O
adhere	NN	O	O
either	NN	O	O
to	NN	O	O
fibronectin-	NN	O	O
or	NN	O	O
laminin-coated	NN	O	O
flasks	NN	O	O
exhibited	NN	O	O
an	NN	O	O
increased	NN	O	O
STAT1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
both	NN	O	O
in	NN	O	O
control	NN	O	O
and	NN	O	O
in	NN	O	O
IFN-gamma-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
events	NN	O	O
leading	NN	O	O
to	NN	O	O
enhanced	NN	O	O
STAT1	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
responsiveness	NN	O	O
concerned	NN	O	O
both	NN	O	O
Y701	NN	O	B-protein
and	NN	O	O
S727	NN	O	B-protein
STAT1	NN	O	I-protein
phosphorylation	NN	O	O
.	NN	O	O

Exogenous	NN	O	O
addition	NN	O	O
of	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
,	NN	O	O
which	NN	O	O
exerts	NN	O	O
an	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
some	NN	O	O
monocytic	NN	O	B-protein
differentiation	NN	O	I-protein
markers	NN	O	I-protein
,	NN	O	O
inhibited	NN	O	O
macrophage	NN	O	O
maturation	NN	O	O
,	NN	O	O
integrin	NN	O	O
expression	NN	O	O
and	NN	O	O
STAT1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
these	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
STAT1	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
pivotal	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
differentiation/maturation	NN	O	O
process	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
as	NN	O	O
an	NN	O	O
early	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
initially	NN	O	O
activated	NN	O	O
by	NN	O	O
adherence	NN	O	O
and	NN	O	O
then	NN	O	O
able	NN	O	O
to	NN	O	O
modulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
functional	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
ICAM-1	NN	O	B-protein
and	NN	O	O
FcgammaRI	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
and	NN	O	O
role	NN	O	O
of	NN	O	O
PML	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
normal	NN	O	O
adult	NN	O	O
hematopoiesis	NN	O	O
:	NN	O	O
functional	NN	O	O
interaction	NN	O	O
between	NN	O	O
PML	NN	O	B-protein
and	NN	O	I-protein
Rb	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
erythropoiesis	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
PML	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
purified	NN	O	O
early	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HPCs	NN	O	B-cell_type
)	NN	O	O
induced	NN	O	O
to	NN	O	O
unilineage	NN	O	O
erythroid	NN	O	O
or	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

PML	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	B-protein
,	NN	O	O
while	NN	O	O
barely	NN	O	O
detectable	NN	O	O
in	NN	O	O
quiescent	NN	O	O
HPCs	NN	O	B-cell_type
,	NN	O	O
are	NN	O	O
consistently	NN	O	O
induced	NN	O	O
by	NN	O	O
growth	NN	O	B-protein
factor	NN	O	I-protein
stimulation	NN	O	O
through	NN	O	O
the	NN	O	O
erythroid	NN	O	O
or	NN	O	O
granulocytic	NN	O	O
lineage	NN	O	O
.	NN	O	O

Thereafter	NN	O	O
,	NN	O	O
PML	NN	O	B-protein
is	NN	O	O
downmodulated	NN	O	O
in	NN	O	O
late	NN	O	O
granulocytic	NN	O	O
maturation	NN	O	O
,	NN	O	O
whereas	NN	O	O
it	NN	O	O
is	NN	O	O
sustainably	NN	O	O
expressed	NN	O	O
through	NN	O	O
the	NN	O	O
erythroid	NN	O	O
pathway	NN	O	O
.	NN	O	O

In	NN	O	O
functional	NN	O	O
studies	NN	O	O
,	NN	O	O
PML	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
antisense	NN	O	O
oligomers	NN	O	O
targeting	NN	O	O
PML	NN	O	B-RNA
mRNA	NN	O	I-RNA
(	NN	O	O
alpha-PML	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
early	NN	O	O
treatment	NN	O	O
(	NN	O	O
day	NN	O	O
0	NN	O	O
HPCs	NN	O	B-cell_type
)	NN	O	O
with	NN	O	O
alpha-PML	NN	O	B-protein
reduced	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
both	NN	O	O
erythroid	NN	O	B-cell_line
and	NN	O	I-cell_line
granulocytic	NN	O	I-cell_line
colonies	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
late	NN	O	O
treatment	NN	O	O
(	NN	O	O
day	NN	O	O
5	NN	O	O
culture	NN	O	O
)	NN	O	O
reduced	NN	O	O
erythroid	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
granulocytic	NN	O	O
,	NN	O	O
clonogenesis	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
PML	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
early	NN	O	O
hematopoiesis	NN	O	O
and	NN	O	O
erythroid	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
granulocytic	NN	O	O
maturation	NN	O	O
.	NN	O	O

The	NN	O	O
pattern	NN	O	O
of	NN	O	O
PML	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
normal	NN	O	O
hematopoiesis	NN	O	O
mimics	NN	O	O
that	NN	O	O
of	NN	O	O
retinoblastoma	NN	O	B-cell_type
pRb	NN	O	B-protein
105	NN	O	I-protein
.	NN	O	O

Combined	NN	O	O
treatment	NN	O	O
of	NN	O	O
HPCs	NN	O	B-cell_type
with	NN	O	O
alpha-PML	NN	O	B-protein
and	NN	O	O
alpha-Rb	NN	O	O
oligomers	NN	O	O
inhibited	NN	O	O
both	NN	O	O
PML	NN	O	O
and	NN	O	O
Rb	NN	O	O
protein	NN	O	O
expression	NN	O	O
and	NN	O	O
completely	NN	O	O
blocked	NN	O	O
erythroid	NN	O	O
colony	NN	O	O
development	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
PML	NN	O	B-protein
and	NN	O	O
pRb	NN	O	B-protein
105	NN	O	I-protein
were	NN	O	O
co-immunoprecipitated	NN	O	O
in	NN	O	O
cellular	NN	O	O
lysates	NN	O	O
derived	NN	O	O
from	NN	O	O
erythroid	NN	O	B-cell_type
precursors	NN	O	I-cell_type
indicating	NN	O	O
that	NN	O	O
this	NN	O	O
functional	NN	O	O
interaction	NN	O	O
may	NN	O	O
have	NN	O	O
a	NN	O	O
biochemical	NN	O	O
basis	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
key	NN	O	O
functional	NN	O	O
role	NN	O	O
of	NN	O	O
PML	NN	O	B-protein
in	NN	O	O
early	NN	O	O
hematopoiesis	NN	O	O
and	NN	O	O
late	NN	O	O
erythropoiesis	NN	O	O
:	NN	O	O
the	NN	O	O
latter	NN	O	O
phenomenon	NN	O	O
may	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
molecular	NN	O	O
and	NN	O	O
functional	NN	O	O
interaction	NN	O	O
of	NN	O	O
PML	NN	O	B-protein
with	NN	O	O
pRb	NN	O	B-protein
105	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Binding	NN	O	O
characteristics	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
multiple	NN	O	O
sclerosis	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
exact	NN	O	O
etiology	NN	O	O
of	NN	O	O
multiple	NN	O	O
sclerosis	NN	O	O
(	NN	O	O
MS	NN	O	O
)	NN	O	O
remains	NN	O	O
unresolved	NN	O	O
,	NN	O	O
immune	NN	O	O
reactions	NN	O	O
are	NN	O	O
believed	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
central	NN	O	O
pathogenic	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

Endogenous	NN	O	O
and	NN	O	O
therapeutic	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
affect	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
,	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
axis	NN	O	O
,	NN	O	O
providing	NN	O	O
evidence	NN	O	O
of	NN	O	O
an	NN	O	O
immune-endocrine	NN	O	O
interplay	NN	O	O
.	NN	O	O

Function	NN	O	O
tests	NN	O	O
in	NN	O	O
MS	NN	O	O
have	NN	O	O
revealed	NN	O	O
dysregulation	NN	O	O
of	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
system	NN	O	O
in	NN	O	O
a	NN	O	O
substantial	NN	O	O
proportion	NN	O	O
of	NN	O	O
patients	NN	O	O
.	NN	O	O

We	NN	O	O
characterized	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
binding	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
39	NN	O	O
MS	NN	O	O
patients	NN	O	O
and	NN	O	O
14	NN	O	O
age-	NN	O	O
and	NN	O	O
sex-matched	NN	O	O
controls	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
and	NN	O	O
binding	NN	O	O
capacity	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
whole-cell	NN	O	O
binding	NN	O	O
assay	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
as	NN	O	O
the	NN	O	O
ligand	NN	O	O
.	NN	O	O

GR	NN	O	B-protein
binding	NN	O	O
parameters	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
significantly	NN	O	O
between	NN	O	O
patients	NN	O	O
(	NN	O	O
Kd	NN	O	O
8.98	NN	O	O
+/-	NN	O	O
1.07	NN	O	O
nM	NN	O	O
,	NN	O	O
Bmax	NN	O	O
183	NN	O	O
+/-	NN	O	O
29.8	NN	O	O
fmol/mg	NN	O	O
)	NN	O	O
and	NN	O	O
controls	NN	O	O
(	NN	O	O
Kd	NN	O	O
9.36	NN	O	O
+/-	NN	O	O
1.17	NN	O	O
nM	NN	O	O
,	NN	O	O
Bmax	NN	O	O
158	NN	O	O
+/-	NN	O	O
16	NN	O	O
fmol/mg	NN	O	O
)	NN	O	O
.	NN	O	O

No	NN	O	O
effect	NN	O	O
of	NN	O	O
age	NN	O	O
,	NN	O	O
sex	NN	O	O
,	NN	O	O
course	NN	O	O
,	NN	O	O
duration	NN	O	O
or	NN	O	O
severity	NN	O	O
of	NN	O	O
disease	NN	O	O
,	NN	O	O
or	NN	O	O
prior	NN	O	O
steroid	NN	O	O
treatments	NN	O	O
was	NN	O	O
detected	NN	O	O
.	NN	O	O

GR	NN	O	B-protein
binding	NN	O	O
parameters	NN	O	O
were	NN	O	O
analyzed	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
the	NN	O	O
combined	NN	O	O
dexamethasone	NN	O	O
-CRH	NN	O	B-protein
test	NN	O	O
,	NN	O	O
which	NN	O	O
reflects	NN	O	O
corticosteroid	NN	O	O
receptor	NN	O	O
function	NN	O	O
at	NN	O	O
the	NN	O	O
hypothalamus	NN	O	O
,	NN	O	O
in	NN	O	O
30	NN	O	O
patients	NN	O	O
and	NN	O	O
9	NN	O	O
controls	NN	O	O
.	NN	O	O

While	NN	O	O
controls	NN	O	O
showed	NN	O	O
a	NN	O	O
moderate	NN	O	O
correlation	NN	O	O
between	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
lymphocytes	NN	O	O
and	NN	O	O
regulatory	NN	O	O
function	NN	O	O
at	NN	O	O
the	NN	O	O
hypothalamic	NN	O	O
level	NN	O	O
,	NN	O	O
the	NN	O	O
patients	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
physiological	NN	O	O
relationship	NN	O	O
between	NN	O	O
binding	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
disturbed	NN	O	O
in	NN	O	O
MS	NN	O	O
.	NN	O	O

-DOCSTART-	O

Phorbol	NN	O	O
ester-induced	NN	O	O
mononuclear	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MEK	NN	O	B-protein
)	NN	O	O
inhibitor	NN	O	O
PD98059	NN	O	O
.	NN	O	O

The	NN	O	O
purpose	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
evaluate	NN	O	O
whether	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
contributes	NN	O	O
to	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
-induced	NN	O	O
mononuclear	NN	O	O
differentiation	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
myeloblastic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
ML-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Upon	NN	O	O
TPA	NN	O	O
treatment	NN	O	O
,	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
ERK1	NN	O	B-protein
and	NN	O	O
ERK2	NN	O	B-protein
rapidly	NN	O	O
increased	NN	O	O
,	NN	O	O
with	NN	O	O
maximal	NN	O	O
induction	NN	O	O
between	NN	O	O
1	NN	O	O
and	NN	O	O
3	NN	O	O
h	NN	O	O
,	NN	O	O
while	NN	O	O
ERK2	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
remained	NN	O	O
constant	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
JNK1	NN	O	B-protein
was	NN	O	O
also	NN	O	O
significantly	NN	O	O
induced	NN	O	O
,	NN	O	O
with	NN	O	O
JNK1	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
increasing	NN	O	O
moderately	NN	O	O
during	NN	O	O
exposure	NN	O	O
to	NN	O	O
TPA	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
PD98059	NN	O	O
,	NN	O	O
a	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MEK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
inhibited	NN	O	O
TPA-induced	NN	O	O
ERK2	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
PD98059	NN	O	O
completely	NN	O	O
blocked	NN	O	O
the	NN	O	O
TPA-induced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
ML-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
features	NN	O	O
associated	NN	O	O
with	NN	O	O
mononuclear	NN	O	O
differentiation	NN	O	O
including	NN	O	O
changes	NN	O	O
in	NN	O	O
morphology	NN	O	O
,	NN	O	O
nonspecific	NN	O	O
esterase	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
phagocytic	NN	O	O
ability	NN	O	O
,	NN	O	O
NADPH	NN	O	B-protein
oxidase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
mitochondrial	NN	O	O
respiration	NN	O	O
,	NN	O	O
and	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
inducibility	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
MEK	NN	O	B-protein
/ERK	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
TPA-induced	NN	O	O
mononuclear	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
and	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Patients	NN	O	O
with	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
do	NN	O	O
not	NN	O	O
display	NN	O	O
cushingoid	NN	O	O
features	NN	O	O
in	NN	O	O
spite	NN	O	O
of	NN	O	O
elevated	NN	O	O
cortisol	NN	O	O
plasma	NN	O	O
levels	NN	O	O
.	NN	O	O

Whether	NN	O	O
a	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
or	NN	O	O
a	NN	O	O
reduced	NN	O	O
availability	NN	O	O
of	NN	O	O
the	NN	O	O
metabolic	NN	O	O
substrates	NN	O	O
necessary	NN	O	O
to	NN	O	O
develop	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
this	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
established	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Twenty-two	NN	O	O
patients	NN	O	O
with	NN	O	O
severe	NN	O	O
restrictive	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
,	NN	O	O
10	NN	O	O
patients	NN	O	O
with	NN	O	O
active	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
,	NN	O	O
and	NN	O	O
24	NN	O	O
healthy	NN	O	O
volunteers	NN	O	O
without	NN	O	O
psychiatric	NN	O	O
disorders	NN	O	O
or	NN	O	O
mood	NN	O	O
alterations	NN	O	O
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	O
receptor	NN	O	O
characteristics	NN	O	O
were	NN	O	O
examined	NN	O	O
on	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
by	NN	O	O
measuring	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
and	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
on	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
,	NN	O	O
which	NN	O	O
represents	NN	O	O
an	NN	O	O
index	NN	O	O
of	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
The	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
MNL	NN	O	B-cell_type
)	NN	O	O
was	NN	O	O
comparable	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
,	NN	O	O
patients	NN	O	O
with	NN	O	O
active	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
,	NN	O	O
and	NN	O	O
normal	NN	O	O
subjects	NN	O	O
(	NN	O	O
binding	NN	O	O
capacity	NN	O	O
3.3	NN	O	O
+/-	NN	O	O
0.23	NN	O	O
vs.	NN	O	O
3.7	NN	O	O
+/-	NN	O	O
0.30	NN	O	O
and	NN	O	O
3.5	NN	O	O
+/-	NN	O	O
0.20	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
)	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
affinity	NN	O	O
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
in	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
Cushing	NN	O	O
's	NN	O	O
patients	NN	O	O
compared	NN	O	O
to	NN	O	O
control	NN	O	O
subjects	NN	O	O
(	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
4.0	NN	O	O
+/-	NN	O	O
0.31	NN	O	O
and	NN	O	O
4.1	NN	O	O
+/-	NN	O	O
0.34	NN	O	O
vs.	NN	O	O
2.9	NN	O	O
+/-	NN	O	O
0.29	NN	O	O
nmol/L	NN	O	O
,	NN	O	O
p	NN	O	O
<	NN	O	O
.001	NN	O	O
)	NN	O	O
and	NN	O	O
inversely	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
urinary	NN	O	O
free	NN	O	O
cortisol	NN	O	O
in	NN	O	O
both	NN	O	O
groups	NN	O	O
of	NN	O	O
patients	NN	O	O
.	NN	O	O

Basal	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
in	NN	O	O
MNL	NN	O	B-cell_type
was	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
in	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
Cushing	NN	O	O
's	NN	O	O
patients	NN	O	O
compared	NN	O	O
to	NN	O	O
control	NN	O	O
subjects	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
.001	NN	O	O
)	NN	O	O
and	NN	O	O
was	NN	O	O
diminished	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
to	NN	O	O
an	NN	O	O
extent	NN	O	O
similar	NN	O	O
to	NN	O	O
control	NN	O	O
subjects	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
,	NN	O	O
but	NN	O	O
significantly	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
.001	NN	O	O
)	NN	O	O
less	NN	O	O
in	NN	O	O
those	NN	O	O
with	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

In	NN	O	O
patients	NN	O	O
with	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
,	NN	O	O
the	NN	O	O
incorporation	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
into	NN	O	O
the	NN	O	O
MNL	NN	O	B-cell_type
was	NN	O	O
inversely	NN	O	O
correlated	NN	O	O
with	NN	O	O
urinary	NN	O	O
free	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
cushingoid	NN	O	O
features	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
is	NN	O	O
not	NN	O	O
ascribable	NN	O	O
to	NN	O	O
a	NN	O	O
reduced	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
but	NN	O	O
is	NN	O	O
more	NN	O	O
likely	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
paucity	NN	O	O
of	NN	O	O
metabolic	NN	O	O
substrates	NN	O	O
.	NN	O	O

-DOCSTART-	O

Different	NN	O	O
sequence	NN	O	O
requirements	NN	O	O
for	NN	O	O
expression	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
within	NN	O	O
a	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
GATA-1	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
lineage-restricted	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
differentiation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
localized	NN	O	O
a	NN	O	O
317	NN	O	O
base	NN	O	O
pair	NN	O	O
cis-acting	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
HS	NN	O	B-DNA
I	NN	O	I-DNA
,	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
hematopoietic-specific	NN	O	B-DNA
DNase	NN	O	I-DNA
I	NN	O	I-DNA
hypersensitive	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
lies	NN	O	O
approx	NN	O	O
.	NN	O	O

3.7	NN	O	O
kilobases	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
hematopoietic-specific	NN	O	I-DNA
GATA-1	NN	O	I-DNA
IE	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

HS	NN	O	B-DNA
I	NN	O	I-DNA
directs	NN	O	O
high-level	NN	O	O
expression	NN	O	O
of	NN	O	O
reporter	NN	O	O
GATA-1/lacZ	NN	O	B-DNA
genes	NN	O	I-DNA
to	NN	O	O
primitive	NN	O	B-cell_type
and	NN	O	I-cell_type
definitive	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
megakaryocytes	NN	O	B-cell_type
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Comparative	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
HS	NN	O	B-DNA
I	NN	O	I-DNA
between	NN	O	O
human	NN	O	O
and	NN	O	O
mouse	NN	O	O
shows	NN	O	O
approx	NN	O	O
.	NN	O	O

63	NN	O	O
%	NN	O	O
nucleotide	NN	O	O
identity	NN	O	O
with	NN	O	O
a	NN	O	O
more	NN	O	O
conserved	NN	O	O
core	NN	O	O
of	NN	O	O
169	NN	O	B-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
(	NN	O	O
86	NN	O	O
%	NN	O	O
identity	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
core	NN	O	O
contains	NN	O	O
a	NN	O	O
GATA	NN	O	B-DNA
site	NN	O	I-DNA
separated	NN	O	O
by	NN	O	O
10	NN	O	O
base	NN	O	O
pairs	NN	O	O
from	NN	O	O
an	NN	O	O
E-box	NN	O	B-DNA
motif	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
composite	NN	O	B-DNA
motif	NN	O	I-DNA
binds	NN	O	O
a	NN	O	O
multi-protein	NN	O	B-protein
hematopoietic-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complex	NN	O	I-protein
which	NN	O	O
includes	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
SCL/tal-1	NN	O	B-protein
,	NN	O	I-protein
E2A	NN	O	B-protein
,	NN	O	O
Lmo2	NN	O	B-protein
and	NN	O	O
Ldb-1	NN	O	B-protein
.	NN	O	O

Point	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
GATA	NN	O	B-DNA
site	NN	O	I-DNA
abolishes	NN	O	O
HS	NN	O	B-DNA
I	NN	O	I-DNA
function	NN	O	O
,	NN	O	O
whereas	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
E-box	NN	O	B-DNA
motif	NN	O	I-DNA
still	NN	O	O
allows	NN	O	O
reporter	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
both	NN	O	O
lineages	NN	O	O
.	NN	O	O

Strict	NN	O	O
dependence	NN	O	O
of	NN	O	O
HS	NN	O	B-DNA
I	NN	O	I-DNA
activity	NN	O	O
on	NN	O	O
a	NN	O	O
GATA	NN	O	B-DNA
site	NN	O	I-DNA
implies	NN	O	O
that	NN	O	O
assembly	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	O
complex	NN	O	O
containing	NN	O	O
a	NN	O	O
GATA-factor	NN	O	B-protein
,	NN	O	O
presumably	NN	O	O
GATA-1	NN	O	B-protein
or	NN	O	O
GATA-2	NN	O	B-protein
,	NN	O	O
is	NN	O	O
critical	NN	O	O
to	NN	O	O
activating	NN	O	O
or	NN	O	O
maintaining	NN	O	O
its	NN	O	O
function	NN	O	O
.	NN	O	O

Further	NN	O	O
dissection	NN	O	O
of	NN	O	O
the	NN	O	O
317	NN	O	B-DNA
base	NN	O	I-DNA
pair	NN	O	I-DNA
region	NN	O	I-DNA
demonstrates	NN	O	O
that	NN	O	O
,	NN	O	O
whereas	NN	O	O
all	NN	O	O
317	NN	O	B-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
expression	NN	O	O
in	NN	O	O
megakaryocytes	NN	O	B-cell_type
,	NN	O	O
only	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
62	NN	O	I-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
are	NN	O	O
needed	NN	O	O
for	NN	O	O
erythroid-specific	NN	O	O
reporter	NN	O	O
expression	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
differential	NN	O	O
lineage	NN	O	O
requirements	NN	O	O
for	NN	O	O
expression	NN	O	O
within	NN	O	O
the	NN	O	O
HS	NN	O	B-DNA
I	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Non-steroidal	NN	O	O
anti-inflammatory	NN	O	O
drugs	NN	O	O
inhibit	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
induce	NN	O	O
HSP70	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
non-steroidal	NN	O	O
anti-inflammatory	NN	O	O
drugs	NN	O	O
(	NN	O	O
NSAIDs	NN	O	O
)	NN	O	O
activate	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
HSF1	NN	O	B-protein
)	NN	O	O
from	NN	O	O
a	NN	O	O
latent	NN	O	B-protein
cytoplasmic	NN	O	I-protein
form	NN	O	I-protein
to	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
,	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
state	NN	O	I-protein
.	NN	O	O

As	NN	O	O
HSF1	NN	O	B-protein
can	NN	O	O
function	NN	O	O
as	NN	O	O
both	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
heat	NN	O	B-DNA
shock	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
repressor	NN	O	O
of	NN	O	O
non-heat	NN	O	B-DNA
shock	NN	O	I-DNA
genes	NN	O	I-DNA
such	NN	O	O
as	NN	O	O
IL1B	NN	O	O
and	NN	O	O
c-	NN	O	O
fos	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
HSF1	NN	O	B-protein
in	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
NSAIDs	NN	O	O
on	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
two	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NSAIDs	NN	O	O
,	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
and	NN	O	O
sulindac	NN	O	O
repress	NN	O	O
the	NN	O	O
IL1B	NN	O	B-DNA
promoter	NN	O	I-DNA
to	NN	O	O
similar	NN	O	O
degree	NN	O	O
to	NN	O	O
heat	NN	O	O
shock	NN	O	O
or	NN	O	O
HSF1	NN	O	B-protein
overexpression	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
and	NN	O	O
additional	NN	O	O
NSAIDs	NN	O	O
used	NN	O	O
at	NN	O	O
concentrations	NN	O	O
that	NN	O	O
activate	NN	O	O
HSF1	NN	O	B-protein
also	NN	O	O
inhibited	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
other	NN	O	O
monocytic	NN	O	B-DNA
genes	NN	O	I-DNA
(	NN	O	O
TNF-alpha	NN	O	B-DNA
,	NN	O	O
IL-1beta	NN	O	B-DNA
,	NN	O	O
IL-6	NN	O	B-DNA
,	NN	O	O
IL-8	NN	O	B-DNA
,	NN	O	O
IL-10	NN	O	B-DNA
,	NN	O	O
ICAM-1	NN	O	B-DNA
)	NN	O	O
activated	NN	O	O
by	NN	O	O
exposure	NN	O	O
to	NN	O	O
a	NN	O	O
pro-inflammatory	NN	O	O
stimulus	NN	O	O
(	NN	O	O
lipopolysaccharide	NN	O	O
,	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

At	NN	O	O
least	NN	O	O
in	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
the	NN	O	O
IL1B	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
repression	NN	O	O
did	NN	O	O
not	NN	O	O
seem	NN	O	O
to	NN	O	O
involve	NN	O	O
another	NN	O	O
factor	NN	O	O
whose	NN	O	O
activity	NN	O	O
is	NN	O	O
affected	NN	O	O
by	NN	O	O
the	NN	O	O
NSAIDs	NN	O	O
,	NN	O	O
NFkappaB	NN	O	B-protein
as	NN	O	O
the	NN	O	O
IL1B	NN	O	B-DNA
promoter	NN	O	I-DNA
fragment	NN	O	I-DNA
used	NN	O	O
in	NN	O	O
our	NN	O	O
studies	NN	O	O
is	NN	O	O
not	NN	O	O
NFkappaB	NN	O	B-protein
responsive	NN	O	O
and	NN	O	O
binds	NN	O	O
specifically	NN	O	O
to	NN	O	O
HSF1	NN	O	B-protein
.	NN	O	O

Exposure	NN	O	O
to	NN	O	O
NSAIDs	NN	O	O
had	NN	O	O
a	NN	O	O
complex	NN	O	O
effect	NN	O	O
on	NN	O	O
HSP	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
while	NN	O	O
sulindac	NN	O	O
activated	NN	O	O
the	NN	O	O
stress	NN	O	O
responsive	NN	O	O
HSP70B	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
only	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
the	NN	O	O
NSAIDs	NN	O	O
induced	NN	O	O
HSP70	NN	O	B-RNA
mRNA	NN	O	I-RNA
species	NN	O	I-RNA
.	NN	O	O

These	NN	O	O
findings	NN	O	O
reflect	NN	O	O
the	NN	O	O
properties	NN	O	O
of	NN	O	O
HSF1	NN	O	B-protein
which	NN	O	O
can	NN	O	O
be	NN	O	O
activated	NN	O	O
to	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
forms	NN	O	I-protein
only	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
activates	NN	O	O
heat	NN	O	B-DNA
shock	NN	O	I-DNA
promoters	NN	O	I-DNA
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
individual	NN	O	O
NSAID	NN	O	O
family	NN	O	O
members	NN	O	O
may	NN	O	O
differentially	NN	O	O
induce	NN	O	O
one	NN	O	O
or	NN	O	O
other	NN	O	O
of	NN	O	O
these	NN	O	O
forms	NN	O	O
.	NN	O	O

Overall	NN	O	O
therefore	NN	O	O
,	NN	O	O
exposure	NN	O	O
to	NN	O	O
NSAIDs	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
profound	NN	O	O
switch	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
with	NN	O	O
suppression	NN	O	O
of	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
macrophage	NN	O	O
activation	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
stress	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
HSF1	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
regulatory	NN	O	O
role	NN	O	O
in	NN	O	O
these	NN	O	O
effects	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Immortalization	NN	O	O
of	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
Tax	NN	O	B-protein
mutants	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
functional	NN	O	B-cell_line
molecular	NN	O	I-cell_line
clone	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
HTLV-1	NN	O	I-protein
)	NN	O	I-protein
transcriptional	NN	O	I-protein
trans-activator	NN	O	I-protein
Tax	NN	O	B-protein
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
have	NN	O	O
transforming	NN	O	O
activity	NN	O	O
in	NN	O	O
multiple	NN	O	B-cell_line
cell	NN	O	I-cell_line
culture	NN	O	I-cell_line
and	NN	O	O
transgenic-mouse	NN	O	O
models	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
activating	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
through	NN	O	O
the	NN	O	O
cyclic	NN	O	B-protein
AMP	NN	O	I-protein
response	NN	O	I-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein/activating	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
CREB/ATF	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
Tax	NN	O	B-protein
activates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
multiple	NN	O	B-DNA
cellular	NN	O	I-DNA
promoters	NN	O	I-DNA
through	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
pathway	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
Tax	NN	O	B-protein
mutants	NN	O	I-protein
M22	NN	O	B-protein
and	NN	O	O
M47	NN	O	B-protein
have	NN	O	O
previously	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
selectively	NN	O	O
abrogate	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
through	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	O
or	NN	O	O
CREB/ATF	NN	O	O
pathway	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

These	NN	O	O
mutations	NN	O	O
were	NN	O	O
introduced	NN	O	O
in	NN	O	O
the	NN	O	O
tax	NN	O	B-DNA
gene	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
ACH	NN	O	B-DNA
functional	NN	O	I-DNA
molecular	NN	O	I-DNA
clone	NN	O	I-DNA
of	NN	O	O
HTLV-1	NN	O	O
,	NN	O	O
and	NN	O	O
virus	NN	O	O
produced	NN	O	O
from	NN	O	O
the	NN	O	O
mutant	NN	O	B-DNA
ACH	NN	O	I-DNA
clones	NN	O	I-DNA
was	NN	O	O
examined	NN	O	O
for	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
replicate	NN	O	O
and	NN	O	O
immortalize	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

While	NN	O	O
virus	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
clone	NN	O	O
containing	NN	O	O
the	NN	O	O
M47	NN	O	B-protein
mutation	NN	O	O
retained	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
immortalize	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
M22	NN	O	B-protein
mutant	NN	O	O
lost	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
immortalize	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
CREB/ATF	NN	O	B-protein
pathway	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
is	NN	O	O
dispensable	NN	O	O
for	NN	O	O
the	NN	O	O
immortalization	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
HTLV-1	NN	O	O
,	NN	O	O
whereas	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
pathway	NN	O	O
may	NN	O	O
be	NN	O	O
critical	NN	O	O
.	NN	O	O

-DOCSTART-	O

Fibroblast	NN	O	B-protein
growth	NN	O	I-protein
factor-1	NN	O	I-protein
(	NN	O	O
FGF-1	NN	O	B-protein
)	NN	O	O
enhances	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
FGF	NN	O	B-cell_line
receptor-bearing	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Fibroblast	NN	O	B-protein
growth	NN	O	I-protein
factors	NN	O	I-protein
(	NN	O	O
FGFs	NN	O	B-protein
)	NN	O	O
are	NN	O	O
heparin-binding	NN	O	B-protein
proteins	NN	O	I-protein
crucial	NN	O	O
to	NN	O	O
embryogenesis	NN	O	O
,	NN	O	O
angiogenesis	NN	O	O
,	NN	O	O
and	NN	O	O
wound	NN	O	O
healing	NN	O	O
.	NN	O	O

FGF-1	NN	O	B-protein
is	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
synovium	NN	O	O
in	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
and	NN	O	O
in	NN	O	O
rejecting	NN	O	O
allografts	NN	O	O
,	NN	O	O
sites	NN	O	O
of	NN	O	O
chronic	NN	O	O
immune-mediated	NN	O	O
inflammation	NN	O	O
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
of	NN	O	O
FGF-1	NN	O	B-protein
-responsive	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
increased	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
these	NN	O	O
disorders	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
high	NN	O	O
percentage	NN	O	O
of	NN	O	O
infiltrating	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
synovium	NN	O	O
express	NN	O	O
receptors	NN	O	O
for	NN	O	O
FGF-1	NN	O	B-protein
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
FGF-1	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
studies	NN	O	O
were	NN	O	O
initiated	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
express	NN	O	O
the	NN	O	O
signaling	NN	O	O
isoform	NN	O	O
of	NN	O	O
FGF	NN	O	B-protein
receptor-1	NN	O	I-protein
.	NN	O	O

These	NN	O	O
experiments	NN	O	O
show	NN	O	O
that	NN	O	O
FGF-1	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
provides	NN	O	O
a	NN	O	O
second	NN	O	O
signal	NN	O	O
that	NN	O	O
augments	NN	O	O
TCR-mediated	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Analogous	NN	O	O
to	NN	O	O
costimulation	NN	O	O
via	NN	O	O
CD28	NN	O	B-protein
,	NN	O	O
this	NN	O	O
activity	NN	O	O
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
Rel/kappaB	NN	O	B-protein
,	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
known	NN	O	O
to	NN	O	O
regulate	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
other	NN	O	O
activation-inducible	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

FGF-1	NN	O	B-protein
alone	NN	O	O
induces	NN	O	O
modest	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
kappaB-binding	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
and	NN	O	O
this	NN	O	O
translocation	NN	O	O
is	NN	O	O
enhanced	NN	O	O
by	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
anti-CD3	NN	O	B-protein
and	NN	O	O
FGF-1	NN	O	B-protein
.	NN	O	O

This	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	I-protein
complex	NN	O	I-protein
is	NN	O	O
composed	NN	O	O
of	NN	O	O
transcriptionally	NN	O	B-protein
active	NN	O	I-protein
p65	NN	O	I-protein
(	NN	O	I-protein
RelA	NN	O	I-protein
)	NN	O	I-protein
/p50	NN	O	I-protein
heterodimers	NN	O	I-protein
and	NN	O	O
results	NN	O	O
primarily	NN	O	O
from	NN	O	O
the	NN	O	O
targeted	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
that	NN	O	O
sequesters	NN	O	O
Rel/kappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
show	NN	O	O
a	NN	O	O
connection	NN	O	O
between	NN	O	O
FGF-1	NN	O	B-protein
signaling	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
outside	NN	O	O
of	NN	O	O
embryonic	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
signaling	NN	O	O
events	NN	O	O
that	NN	O	O
link	NN	O	O
FGF	NN	O	B-protein
receptor-1	NN	O	I-protein
engagement	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
are	NN	O	O
probably	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
costimulation	NN	O	O
pathway	NN	O	O
,	NN	O	O
since	NN	O	O
FGF-1	NN	O	B-protein
-induced	NN	O	O
Rel/kappaB	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
do	NN	O	O
not	NN	O	O
contain	NN	O	O
significant	NN	O	O
levels	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	O
are	NN	O	O
not	NN	O	O
identical	NN	O	O
with	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
response	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Escape	NN	O	O
of	NN	O	O
human	NN	O	O
cytomegalovirus	NN	O	O
from	NN	O	O
HLA-DR	NN	O	B-protein
-restricted	NN	O	O
CD4	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
T-cell	NN	O	O
response	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
repression	NN	O	O
of	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
-induced	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Human	NN	O	O
cytomegalovirus	NN	O	O
(	NN	O	O
HCMV	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
betaherpesvirus	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
pathogen	NN	O	O
which	NN	O	O
escapes	NN	O	O
immune	NN	O	O
recognition	NN	O	O
through	NN	O	O
various	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
HCMV	NN	O	O
down	NN	O	O
regulates	NN	O	O
gamma	NN	O	O
interferon	NN	O	O
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
-induced	NN	O	O
HLA-DR	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
U373	NN	O	B-cell_line
MG	NN	O	I-cell_line
astrocytoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
due	NN	O	O
to	NN	O	O
a	NN	O	O
defect	NN	O	O
downstream	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
phosphorylation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

Repression	NN	O	O
of	NN	O	O
class	NN	O	O
II	NN	O	O
transactivator	NN	O	O
(	NN	O	O
CIITA	NN	O	B-DNA
)	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
is	NN	O	O
detected	NN	O	O
within	NN	O	O
the	NN	O	O
first	NN	O	O
hours	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
-HCMV	NN	O	O
coincubation	NN	O	O
and	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
synthesis	NN	O	O
.	NN	O	O

This	NN	O	O
defect	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
presentation	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-protein
immediate-early	NN	O	I-protein
protein	NN	O	I-protein
IE1	NN	O	B-protein
to	NN	O	O
specific	NN	O	O
CD4	NN	O	B-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
when	NN	O	O
U373	NN	O	B-cell_line
MG	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
used	NN	O	O
as	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
are	NN	O	O
treated	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
plus	NN	O	O
HCMV	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
presentation	NN	O	O
of	NN	O	O
endogenously	NN	O	O
synthesized	NN	O	O
IE1	NN	O	B-protein
can	NN	O	O
be	NN	O	O
restored	NN	O	O
when	NN	O	O
U373	NN	O	B-cell_line
MG	NN	O	I-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
transfected	NN	O	O
with	NN	O	O
CIITA	NN	O	B-DNA
prior	NN	O	O
to	NN	O	O
infection	NN	O	O
with	NN	O	O
HCMV	NN	O	O
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
the	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
defect	NN	O	O
induced	NN	O	O
by	NN	O	O
HCMV	NN	O	O
resides	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-gamma-responsive	NN	O	B-DNA
promoter	NN	O	I-DNA
of	NN	O	O
CIITA	NN	O	B-DNA
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
demonstration	NN	O	O
of	NN	O	O
a	NN	O	O
viral	NN	O	O
inhibition	NN	O	O
of	NN	O	O
CIITA	NN	O	B-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

3-deazaadenosine	NN	O	O
,	NN	O	O
a	NN	O	O
S-adenosylhomocysteine	NN	O	B-protein
hydrolase	NN	O	I-protein
inhibitor	NN	O	O
,	NN	O	O
has	NN	O	O
dual	NN	O	O
effects	NN	O	O
on	NN	O	O
NF-kappaB	NN	O	B-protein
regulation	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
and	NN	O	O
promotion	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
degradation	NN	O	O
.	NN	O	O

Previously	NN	O	O
we	NN	O	O
reported	NN	O	O
that	NN	O	O
3-deazaadenosine	NN	O	O
(	NN	O	O
DZA	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitor	NN	O	O
and	NN	O	O
substrate	NN	O	O
for	NN	O	O
S-adenosylhomocysteine	NN	O	B-protein
hydrolase	NN	O	I-protein
inhibits	NN	O	O
bacterial	NN	O	O
lipopolysaccharide-induced	NN	O	O
transcription	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
and	NN	O	O
interleukin-1beta	NN	O	B-protein
in	NN	O	O
mouse	NN	O	B-cell_line
macrophage	NN	O	I-cell_line
RAW	NN	O	I-cell_line
264.7	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
DZA	NN	O	O
on	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
regulation	NN	O	O
.	NN	O	O

DZA	NN	O	O
inhibits	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
through	NN	O	O
the	NN	O	O
hindrance	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
(	NN	O	O
Rel-A	NN	O	B-protein
)	NN	O	O
phosphorylation	NN	O	O
without	NN	O	O
reduction	NN	O	O
of	NN	O	O
its	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
DZA	NN	O	O
on	NN	O	O
NF-kappaB	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
is	NN	O	O
potentiated	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
homocysteine	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
DZA	NN	O	O
promotes	NN	O	O
the	NN	O	O
proteolytic	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IkappaBbeta	NN	O	B-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
an	NN	O	O
increase	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
its	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
RAW	NN	O	B-cell_line
264.7	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
reduction	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
by	NN	O	O
DZA	NN	O	O
is	NN	O	O
neither	NN	O	O
involved	NN	O	O
in	NN	O	O
IkappaB	NN	O	B-protein
kinase	NN	O	I-protein
complex	NN	O	I-protein
activation	NN	O	O
nor	NN	O	O
modulated	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
homocysteine	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
strongly	NN	O	O
suggests	NN	O	O
that	NN	O	O
DZA	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
potent	NN	O	O
drug	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
diseases	NN	O	O
in	NN	O	O
which	NN	O	O
NF-kappaB	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
central	NN	O	O
pathogenic	NN	O	O
role	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
useful	NN	O	O
tool	NN	O	O
for	NN	O	O
studying	NN	O	O
the	NN	O	O
regulation	NN	O	O
and	NN	O	O
physiological	NN	O	O
functions	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
gamma/delta	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
receptor	NN	O	I-cell_type
positive	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
pregnancy	NN	O	O
.	NN	O	O

PROBLEM	NN	O	O
:	NN	O	O
Due	NN	O	O
to	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
classical	NN	O	B-protein
HLA	NN	O	I-protein
antigens	NN	O	I-protein
on	NN	O	O
the	NN	O	O
trophoblast	NN	O	O
,	NN	O	O
fetal	NN	O	B-protein
antigens	NN	O	I-protein
are	NN	O	O
possibly	NN	O	O
presented	NN	O	O
in	NN	O	O
a	NN	O	O
non	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	O
MHC	NN	O	B-protein
)	NN	O	O
restricted	NN	O	O
way	NN	O	O
.	NN	O	O

Decidual	NN	O	B-cell_type
gammadelta	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
significantly	NN	O	O
increase	NN	O	O
in	NN	O	O
number	NN	O	O
during	NN	O	O
pregnancy	NN	O	O
,	NN	O	O
might	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
recognition	NN	O	O
of	NN	O	O
fetal	NN	O	B-protein
antigens	NN	O	I-protein
and	NN	O	O
also	NN	O	O
in	NN	O	O
determining	NN	O	O
the	NN	O	O
quality	NN	O	O
of	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
these	NN	O	O
antigens	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
study	NN	O	O
was	NN	O	O
aimed	NN	O	O
at	NN	O	O
investigating	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
cell	NN	O	O
population	NN	O	O
in	NN	O	O
progesterone-dependent	NN	O	O
immunomodulation	NN	O	O
.	NN	O	O

METHOD	NN	O	O
OF	NN	O	O
STUDY	NN	O	O
:	NN	O	O
Peripheral	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
healthy	NN	O	O
pregnant	NN	O	O
women	NN	O	O
and	NN	O	O
from	NN	O	O
habitual	NN	O	O
aborters	NN	O	O
were	NN	O	O
tested	NN	O	O
by	NN	O	O
immunocytochemistry	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
gamma/delta	NN	O	O
T	NN	O	O
cell	NN	O	O
receptor	NN	O	O
(	NN	O	O
TCR	NN	O	O
)	NN	O	O
and	NN	O	O
progesterone	NN	O	O
receptor	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
treatment	NN	O	O
with	NN	O	O
a	NN	O	O
pan	NN	O	O
anti	NN	O	B-protein
gamma/delta	NN	O	I-protein
antibody	NN	O	I-protein
,	NN	O	O
lymphocytes	NN	O	B-cell_type
were	NN	O	O
incubated	NN	O	O
for	NN	O	O
3	NN	O	O
hr	NN	O	O
with	NN	O	O
the	NN	O	O
antibody	NN	O	O
,	NN	O	O
and	NN	O	O
then	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-10	NN	O	I-protein
,	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
progesterone-induced	NN	O	B-protein
blocking	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
PIBF	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
(	NN	O	O
by	NN	O	O
immuno-cytochemistry	NN	O	O
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
were	NN	O	O
determined	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
In	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
healthy	NN	O	O
pregnant	NN	O	O
women	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
gamma/delta	NN	O	B-cell_type
TCR+	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
than	NN	O	O
in	NN	O	O
that	NN	O	O
of	NN	O	O
recurrent	NN	O	O
aborters	NN	O	O
or	NN	O	O
of	NN	O	O
non-pregnant	NN	O	O
individuals	NN	O	O
.	NN	O	O

Ninety-seven	NN	O	O
percent	NN	O	O
of	NN	O	O
gamma/delta	NN	O	B-cell_type
TCR+	NN	O	I-cell_type
pregnancy	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
expressed	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
a	NN	O	O
specific	NN	O	O
antibody	NN	O	O
to	NN	O	O
the	NN	O	O
gamma/delta	NN	O	O
TCR	NN	O	O
inhibited	NN	O	O
PIBF	NN	O	O
-as	NN	O	O
well	NN	O	O
as	NN	O	O
IL-10	NN	O	O
production	NN	O	O
,	NN	O	O
whereas	NN	O	O
it	NN	O	O
increased	NN	O	O
NK	NN	O	O
activity	NN	O	O
and	NN	O	O
IL-12	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
These	NN	O	O
data	NN	O	O
suggest	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
gamma/delta	NN	O	B-cell_type
TCR-bearing	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
progesterone-dependent	NN	O	O
immunomodulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Monocyte	NN	O	O
adhesion	NN	O	O
and	NN	O	O
spreading	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
Rho	NN	O	B-protein
-regulated	NN	O	O
receptor	NN	O	O
clustering	NN	O	O
.	NN	O	O

The	NN	O	O
GTPase	NN	O	B-protein
Rho	NN	O	B-protein
is	NN	O	O
known	NN	O	O
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
assembly	NN	O	O
of	NN	O	O
integrin-containing	NN	O	O
focal	NN	O	O
adhesions	NN	O	O
and	NN	O	O
actin	NN	O	O
stress	NN	O	O
fibers	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Rho	NN	O	B-protein
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte-binding	NN	O	B-protein
receptors	NN	O	I-protein
E-selectin	NN	O	B-protein
,	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
VCAM-1	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
Rho	NN	O	B-protein
activity	NN	O	O
with	NN	O	O
C3	NN	O	B-protein
transferase	NN	O	I-protein
or	NN	O	O
N19RhoA	NN	O	B-protein
,	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
RhoA	NN	O	B-protein
mutant	NN	O	I-protein
,	NN	O	O
reduced	NN	O	O
the	NN	O	O
adhesion	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
to	NN	O	O
activated	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
inhibited	NN	O	O
their	NN	O	O
spreading	NN	O	O
.	NN	O	O

Similar	NN	O	O
effects	NN	O	O
were	NN	O	O
observed	NN	O	O
after	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
cytochalasin	NN	O	O
D	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
Rac	NN	O	I-protein
and	NN	O	I-protein
Cdc42	NN	O	I-protein
proteins	NN	O	I-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
or	NN	O	O
spreading	NN	O	O
.	NN	O	O

C3	NN	O	O
transferase	NN	O	O
and	NN	O	O
cytochalasin	NN	O	O
D	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
expression	NN	O	O
levels	NN	O	O
of	NN	O	O
monocyte-binding	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
did	NN	O	O
inhibit	NN	O	O
clustering	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
VCAM-1	NN	O	B-protein
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
induced	NN	O	O
by	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
or	NN	O	O
cross-linking	NN	O	B-protein
antibodies	NN	O	I-protein
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
N19RhoA	NN	O	B-protein
inhibited	NN	O	O
receptor	NN	O	O
clustering	NN	O	O
.	NN	O	O

Monocyte	NN	O	O
adhesion	NN	O	O
and	NN	O	O
receptor	NN	O	O
cross-linking	NN	O	O
induced	NN	O	O
stress	NN	O	O
fiber	NN	O	O
assembly	NN	O	O
,	NN	O	O
and	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
myosin	NN	O	B-protein
light	NN	O	I-protein
chain	NN	O	I-protein
kinase	NN	O	I-protein
prevented	NN	O	O
this	NN	O	O
response	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
receptor	NN	O	O
clustering	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
receptor	NN	O	O
clusters	NN	O	O
colocalized	NN	O	O
with	NN	O	O
ezrin/moesin/radixin	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
Rho	NN	O	B-protein
is	NN	O	O
required	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
the	NN	O	O
assembly	NN	O	O
of	NN	O	O
stable	NN	O	O
adhesions	NN	O	O
with	NN	O	O
monocytes	NN	O	B-cell_type
via	NN	O	O
the	NN	O	O
clustering	NN	O	O
of	NN	O	O
monocyte-binding	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
their	NN	O	O
association	NN	O	O
with	NN	O	O
the	NN	O	O
actin	NN	O	O
cytoskeleton	NN	O	O
,	NN	O	O
independent	NN	O	O
of	NN	O	O
stress	NN	O	O
fiber	NN	O	O
formation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Selection	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
repertoire	NN	O	O
.	NN	O	O

Advances	NN	O	O
in	NN	O	O
gene	NN	O	O
technology	NN	O	O
have	NN	O	O
allowed	NN	O	O
the	NN	O	O
manipulation	NN	O	O
of	NN	O	O
molecular	NN	O	O
interactions	NN	O	O
that	NN	O	O
shape	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
repertoire	NN	O	O
.	NN	O	O

Although	NN	O	O
recognized	NN	O	O
as	NN	O	O
fundamental	NN	O	O
aspects	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
development	NN	O	O
,	NN	O	O
only	NN	O	O
recently	NN	O	O
have	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
governing	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	O
selection	NN	O	O
been	NN	O	O
examined	NN	O	O
at	NN	O	O
a	NN	O	O
molecular	NN	O	O
level	NN	O	O
.	NN	O	O

Positive	NN	O	O
selection	NN	O	O
refers	NN	O	O
to	NN	O	O
the	NN	O	O
active	NN	O	O
process	NN	O	O
of	NN	O	O
rescuing	NN	O	O
MHC-restricted	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
from	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Negative	NN	O	O
selection	NN	O	O
refers	NN	O	O
to	NN	O	O
the	NN	O	O
deletion	NN	O	O
or	NN	O	O
inactivation	NN	O	O
of	NN	O	O
potentially	NN	O	O
autoreactive	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
review	NN	O	O
focuses	NN	O	O
on	NN	O	O
interactions	NN	O	O
during	NN	O	O
thymocyte	NN	O	B-cell_type
maturation	NN	O	O
that	NN	O	O
define	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
repertoire	NN	O	O
,	NN	O	O
with	NN	O	O
an	NN	O	O
emphasis	NN	O	O
placed	NN	O	O
on	NN	O	O
current	NN	O	O
literature	NN	O	O
within	NN	O	O
this	NN	O	O
field	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD28	NN	O	B-protein
costimulation	NN	O	O
augments	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
of	NN	O	O
activated	NN	O	O
lamina	NN	O	O
propria	NN	O	O
T	NN	O	O
cells	NN	O	O
by	NN	O	O
increasing	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
without	NN	O	O
enhancing	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
transactivation	NN	O	O
.	NN	O	O

The	NN	O	O
pathways	NN	O	O
leading	NN	O	O
to	NN	O	O
activation	NN	O	O
in	NN	O	O
lamina	NN	O	B-cell_type
propria	NN	O	I-cell_type
(	NN	O	I-cell_type
LP	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
different	NN	O	O
from	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

LP	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
exhibit	NN	O	O
enhanced	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
when	NN	O	O
activated	NN	O	O
through	NN	O	O
the	NN	O	O
CD2	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

Coligation	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
synergistic	NN	O	O
enhancement	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
characterized	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
augmentation	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
an	NN	O	O
IL-2	NN	O	B-protein
promoter	NN	O	O
CD28	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CD28RE	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
along	NN	O	O
with	NN	O	O
enhanced	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
characterized	NN	O	O
molecular	NN	O	O
events	NN	O	O
involved	NN	O	O
in	NN	O	O
CD28	NN	O	B-protein
costimulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
in	NN	O	O
LP	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	I-cell_type
LPMC	NN	O	I-cell_type
)	NN	O	I-cell_type
.	NN	O	O

LPMC	NN	O	B-cell_type
exhibited	NN	O	O
increased	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
CD28	NN	O	B-protein
costimulation	NN	O	O
,	NN	O	O
compared	NN	O	O
with	NN	O	O
cells	NN	O	O
activated	NN	O	O
through	NN	O	O
CD2	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

IL-2	NN	O	B-protein
secretion	NN	O	O
was	NN	O	O
paralleled	NN	O	O
by	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
resulting	NN	O	O
from	NN	O	O
enhanced	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
stability	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
,	NN	O	O
EMSA	NN	O	O
revealed	NN	O	O
that	NN	O	O
CD28	NN	O	B-protein
coligation	NN	O	O
of	NN	O	O
CD2-activated	NN	O	B-cell_type
LPMC	NN	O	I-cell_type
does	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
increased	NN	O	O
binding	NN	O	O
of	NN	O	O
trans-factors	NN	O	O
to	NN	O	O
the	NN	O	O
CD28RE	NN	O	B-DNA
,	NN	O	O
nor	NN	O	O
did	NN	O	O
Western	NN	O	O
blots	NN	O	O
detect	NN	O	O
changes	NN	O	O
in	NN	O	O
I-kappaBalpha	NN	O	B-protein
or	NN	O	O
I-kappaBbeta	NN	O	B-protein
levels	NN	O	O
following	NN	O	O
CD28	NN	O	B-protein
coligation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
CD28	NN	O	B-protein
coligation	NN	O	O
fails	NN	O	O
to	NN	O	O
enhance	NN	O	O
IL-2	NN	O	B-DNA
promoter-reporter	NN	O	I-DNA
or	NN	O	O
RE/AP	NN	O	B-protein
construct	NN	O	O
expression	NN	O	O
in	NN	O	O
CD2-activated	NN	O	B-cell_type
LPMC	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
results	NN	O	O
reported	NN	O	O
herein	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
CD28	NN	O	B-protein
cosignaling	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
in	NN	O	O
LP	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
unique	NN	O	O
to	NN	O	O
that	NN	O	O
compartment	NN	O	O
and	NN	O	O
differ	NN	O	O
from	NN	O	O
those	NN	O	O
seen	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
a	NN	O	O
biological	NN	O	O
significance	NN	O	O
for	NN	O	O
different	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
initiation	NN	O	O
and	NN	O	O
maintenance	NN	O	O
of	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
repertoire	NN	O	I-protein
found	NN	O	O
in	NN	O	O
the	NN	O	O
mucosa	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
Th1	NN	O	B-protein
and	NN	O	I-protein
Th2	NN	O	I-protein
type	NN	O	I-protein
cytokines	NN	O	I-protein
responding	NN	O	O
to	NN	O	O
HBsAg	NN	O	B-protein
and	NN	O	O
HBxAg	NN	O	B-protein
in	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
cytokine	NN	O	B-protein
pattern	NN	O	I-protein
on	NN	O	O
viral	NN	O	O
antigen	NN	O	O
recognition	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
exert	NN	O	O
a	NN	O	O
profound	NN	O	O
influence	NN	O	O
on	NN	O	O
the	NN	O	O
resolution	NN	O	O
of	NN	O	O
viral	NN	O	O
infections	NN	O	O
and	NN	O	O
viral	NN	O	O
clearance	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
was	NN	O	O
initiated	NN	O	O
to	NN	O	O
investigate	NN	O	O
whether	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
imbalance	NN	O	O
oriented	NN	O	O
toward	NN	O	O
Th2	NN	O	O
type	NN	O	O
response	NN	O	O
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
.	NN	O	O

Cytokine	NN	O	B-protein
profiles	NN	O	I-protein
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
associated	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
were	NN	O	O
analysed	NN	O	O
by	NN	O	O
RT-PCR	NN	O	O
.	NN	O	O

Upon	NN	O	O
HBsAg	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-10	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
41	NN	O	O
%	NN	O	O
,	NN	O	O
8	NN	O	O
%	NN	O	O
,	NN	O	O
41	NN	O	O
%	NN	O	O
,	NN	O	O
and	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Among	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
was	NN	O	O
associated	NN	O	O
with	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
serum	NN	O	B-protein
AST/ALT	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
could	NN	O	O
not	NN	O	O
prove	NN	O	O
that	NN	O	O
Th2	NN	O	B-protein
type	NN	O	I-protein
cytokines	NN	O	I-protein
had	NN	O	O
a	NN	O	O
protective	NN	O	O
effect	NN	O	O
on	NN	O	O
hepatocytes	NN	O	B-cell_type
.	NN	O	O

Upon	NN	O	O
HBxAg	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
recognizable	NN	O	O
association	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
patterns	NN	O	I-protein
with	NN	O	O
AST/ALT	NN	O	O
levels	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
production	NN	O	O
of	NN	O	O
a	NN	O	O
Th1	NN	O	B-protein
cytokine	NN	O	I-protein
,	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
by	NN	O	O
HBsAg	NN	O	B-protein
-reactive	NN	O	O
cells	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
hepatocyte	NN	O	O
damage	NN	O	O
in	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
,	NN	O	O
while	NN	O	O
no	NN	O	O
counteracting	NN	O	O
effect	NN	O	O
of	NN	O	O
Th2	NN	O	B-protein
cytokines	NN	O	I-protein
produced	NN	O	O
by	NN	O	O
those	NN	O	O
cells	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Apoptosis	NN	O	O
and	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
in	NN	O	O
malignant	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
arsenic	NN	O	O
trioxide	NN	O	O
at	NN	O	O
clinically	NN	O	O
achievable	NN	O	O
concentrations	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Arsenic	NN	O	O
trioxide	NN	O	O
(	NN	O	O
As2O3	NN	O	O
)	NN	O	O
can	NN	O	O
induce	NN	O	O
clinical	NN	O	O
remission	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
via	NN	O	O
induction	NN	O	O
of	NN	O	O
differentiation	NN	O	O
and	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
(	NN	O	O
apoptosis	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
As2O3	NN	O	O
on	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
malignant	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
growth-inhibitory	NN	O	O
and	NN	O	O
apoptotic	NN	O	O
effects	NN	O	O
of	NN	O	O
As2O3	NN	O	O
can	NN	O	O
be	NN	O	O
observed	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
at	NN	O	O
clinically	NN	O	O
achievable	NN	O	O
concentrations	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Eight	NN	O	O
malignant	NN	O	B-cell_line
lymphocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
primary	NN	O	B-cell_line
cultures	NN	O	I-cell_line
of	NN	O	O
lymphocytic	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
and	NN	O	I-cell_type
lymphoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
treated	NN	O	O
with	NN	O	O
As2O3	NN	O	O
,	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
dithiothreitol	NN	O	O
(	NN	O	O
DTT	NN	O	O
)	NN	O	O
or	NN	O	O
buthionine	NN	O	O
sulfoximine	NN	O	O
(	NN	O	O
BSO	NN	O	O
)	NN	O	O
(	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
glutathione	NN	O	O
synthesis	NN	O	O
)	NN	O	O
.	NN	O	O

Apoptosis	NN	O	O
was	NN	O	O
assessed	NN	O	O
by	NN	O	O
cell	NN	O	O
morphology	NN	O	O
,	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
,	NN	O	O
annexin	NN	O	O
V	NN	O	O
protein	NN	O	O
level	NN	O	O
,	NN	O	O
and	NN	O	O
terminal	NN	O	B-protein
deoxynucleotidyl	NN	O	I-protein
transferase	NN	O	I-protein
labeling	NN	O	O
of	NN	O	O
DNA	NN	O	B-DNA
fragments	NN	O	I-DNA
.	NN	O	O

Cellular	NN	O	O
proliferation	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
5-bromo-2'-deoxyuridine	NN	O	O
incorporation	NN	O	O
into	NN	O	O
DNA	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
and	NN	O	O
by	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
mitotic	NN	O	O
arrest	NN	O	O
assay	NN	O	O
.	NN	O	O

Mitochondrial	NN	O	O
transmembrane	NN	O	O
potential	NN	O	O
(	NN	O	O
delta	NN	O	O
psi	NN	O	O
(	NN	O	O
m	NN	O	O
)	NN	O	O
)	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
rhodamine	NN	O	O
123	NN	O	O
staining	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

Protein	NN	O	O
expression	NN	O	O
was	NN	O	O
assessed	NN	O	O
by	NN	O	O
western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
or	NN	O	O
immunofluorescence	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Therapeutic	NN	O	O
concentrations	NN	O	O
of	NN	O	O
As2O3	NN	O	O
(	NN	O	O
1-2	NN	O	O
microM	NN	O	O
)	NN	O	O
had	NN	O	O
dual	NN	O	O
effects	NN	O	O
on	NN	O	O
malignant	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
1	NN	O	O
)	NN	O	O
inhibition	NN	O	O
of	NN	O	O
growth	NN	O	O
through	NN	O	O
adenosine	NN	O	O
triphosphate	NN	O	O
(	NN	O	O
ATP	NN	O	O
)	NN	O	O
depletion	NN	O	O
and	NN	O	O
prolongation	NN	O	O
of	NN	O	O
cell	NN	O	O
cycle	NN	O	O
time	NN	O	O
and	NN	O	O
2	NN	O	O
)	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

As2O3-induced	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
preceded	NN	O	O
by	NN	O	O
delta	NN	O	O
psi	NN	O	O
(	NN	O	O
m	NN	O	O
)	NN	O	O
collapse	NN	O	O
.	NN	O	O

DTT	NN	O	O
antagonized	NN	O	O
and	NN	O	O
BSO	NN	O	O
enhanced	NN	O	O
As2O3-induced	NN	O	O
ATP	NN	O	O
depletion	NN	O	O
,	NN	O	O
delta	NN	O	O
psi	NN	O	O
(	NN	O	O
m	NN	O	O
)	NN	O	O
collapse	NN	O	O
,	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Caspase-3	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
usually	NN	O	O
resulting	NN	O	O
from	NN	O	O
delta	NN	O	O
psi	NN	O	O
(	NN	O	O
m	NN	O	O
)	NN	O	O
collapse	NN	O	O
,	NN	O	O
was	NN	O	O
not	NN	O	O
always	NN	O	O
associated	NN	O	O
with	NN	O	O
As2O3-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

As2O3	NN	O	O
induced	NN	O	O
PML	NN	O	O
(	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
)	NN	O	O
protein	NN	O	O
degradation	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
modulate	NN	O	O
expression	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
cycle-related	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
including	NN	O	O
c-myc	NN	O	B-protein
,	NN	O	O
retinoblastoma	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
cyclin-dependent	NN	O	B-protein
kinase	NN	O	I-protein
4	NN	O	I-protein
,	NN	O	O
cyclin	NN	O	B-protein
D1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
p53	NN	O	B-protein
,	NN	O	O
or	NN	O	O
expression	NN	O	O
of	NN	O	O
differentiation-related	NN	O	B-protein
antigens	NN	O	I-protein
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Substantial	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
without	NN	O	O
evidence	NN	O	O
of	NN	O	O
differentiation	NN	O	O
were	NN	O	O
induced	NN	O	O
in	NN	O	O
most	NN	O	O
malignant	NN	O	B-cell_type
lymphocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
1-2	NN	O	O
microM	NN	O	O
As2O3	NN	O	O
.	NN	O	O

As2O3	NN	O	O
may	NN	O	O
prove	NN	O	O
useful	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
malignant	NN	O	O
lymphoproliferative	NN	O	O
disorders	NN	O	O
.	NN	O	O

-DOCSTART-	O

Control	NN	O	O
of	NN	O	O
cell	NN	O	O
cycle	NN	O	O
entry	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
infected	NN	O	O
by	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
.	NN	O	O

Infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
and	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

To	NN	O	O
interpret	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
EBV	NN	O	O
activates	NN	O	O
the	NN	O	O
cell	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
assayed	NN	O	O
many	NN	O	O
proteins	NN	O	O
involved	NN	O	O
in	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
G0	NN	O	O
and	NN	O	O
G1	NN	O	O
phases	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
EBV	NN	O	O
infection	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
changes	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
early	NN	O	O
induction	NN	O	O
of	NN	O	O
cyclin	NN	O	B-protein
D2	NN	O	I-protein
,	NN	O	O
are	NN	O	O
dependent	NN	O	O
on	NN	O	O
expression	NN	O	O
of	NN	O	O
EBV	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
an	NN	O	O
alteration	NN	O	O
in	NN	O	O
the	NN	O	O
E2F-4	NN	O	B-protein
profile	NN	O	O
was	NN	O	O
partly	NN	O	O
independent	NN	O	O
of	NN	O	O
viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
presumably	NN	O	O
occurring	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
signal	NN	O	O
transduction	NN	O	O
activated	NN	O	O
when	NN	O	O
the	NN	O	O
virus	NN	O	O
binds	NN	O	O
to	NN	O	O
its	NN	O	O
receptor	NN	O	O
,	NN	O	O
CD21	NN	O	B-protein
.	NN	O	O

By	NN	O	O
comparing	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
controlling	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
including	NN	O	O
those	NN	O	O
encoding	NN	O	O
several	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
BCL-2	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
proteins	NN	O	O
,	NN	O	O
the	NN	O	O
known	NN	O	O
relative	NN	O	O
resistance	NN	O	O
of	NN	O	O
EBV-immortalized	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
to	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
low	NN	O	O
serum	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
correlate	NN	O	O
with	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
BCL-2	NN	O	B-protein
and	NN	O	O
A20	NN	O	B-protein
.	NN	O	O

A20	NN	O	O
can	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
LMP-1	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Quantitative	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
direct	NN	O	O
temporal	NN	O	O
relationship	NN	O	O
between	NN	O	O
LMP-1	NN	O	B-protein
protein	NN	O	I-protein
levels	NN	O	O
and	NN	O	O
active	NN	O	O
NF-kappaB	NN	O	B-protein
during	NN	O	O
the	NN	O	O
time	NN	O	O
course	NN	O	O
of	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

CTLA-4	NN	O	B-protein
-Mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
early	NN	O	O
events	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

CTLA-4	NN	O	B-protein
engagement	NN	O	O
by	NN	O	O
mAbs	NN	O	B-protein
inhibits	NN	O	O
,	NN	O	O
while	NN	O	O
CD28	NN	O	B-protein
enhances	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
and	NN	O	O
proliferation	NN	O	O
upon	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
CTLA-4	NN	O	B-protein
-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
of	NN	O	O
naive	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
Ab	NN	O	O
cross-linking	NN	O	O
.	NN	O	O

CTLA-4	NN	O	B-protein
ligation	NN	O	O
inhibited	NN	O	O
CD3/	NN	O	O
CD28	NN	O	B-protein
-induced	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
,	NN	O	O
which	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
in	NN	O	O
part	NN	O	O
through	NN	O	O
decreasing	NN	O	O
NF-AT	NN	O	B-protein
accumulation	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
CTLA-4	NN	O	B-protein
ligation	NN	O	O
did	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
affect	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
-mediated	NN	O	O
stabilization	NN	O	O
of	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Further	NN	O	O
,	NN	O	O
CTLA-4	NN	O	B-protein
engagement	NN	O	O
inhibited	NN	O	O
progression	NN	O	O
through	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
cyclin	NN	O	B-protein
D3	NN	O	I-protein
,	NN	O	O
cyclin-dependent	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	B-protein
cdk	NN	O	I-protein
)	NN	O	I-protein
4	NN	O	I-protein
,	NN	O	O
and	NN	O	O
cdk6	NN	O	B-protein
when	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
anti-CD3/CD28	NN	O	B-protein
and	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
CTLA-4	NN	O	B-protein
signaling	NN	O	O
inhibits	NN	O	O
events	NN	O	O
early	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
both	NN	O	O
at	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
and	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
-independent	NN	O	O
events	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
and	NN	O	O
does	NN	O	O
not	NN	O	O
simply	NN	O	O
oppose	NN	O	O
CD28	NN	O	B-protein
-mediated	NN	O	O
costimulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
cooperates	NN	O	O
with	NN	O	O
the	NN	O	O
erythropoietin	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
c-Kit	NN	O	B-protein
to	NN	O	O
enhance	NN	O	O
and	NN	O	O
sustain	NN	O	O
proliferation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Although	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
Epo	NN	O	B-protein
)	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
mature	NN	O	B-cell_type
red	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
cooperation	NN	O	O
with	NN	O	O
other	NN	O	O
factors	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
a	NN	O	O
proper	NN	O	O
balance	NN	O	O
between	NN	O	O
progenitor	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
avian	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
,	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
cooperate	NN	O	O
with	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
receptors	NN	O	I-protein
to	NN	O	O
induce	NN	O	O
renewal	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
corticosteroids	NN	O	O
in	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
expansion	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
liquid	NN	O	O
cultures	NN	O	O
and	NN	O	O
colony	NN	O	O
assays	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
(	NN	O	O
Dex	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
synthetic	NN	O	O
glucocorticoid	NN	O	O
hormone	NN	O	O
,	NN	O	O
cooperated	NN	O	O
with	NN	O	O
Epo	NN	O	B-protein
and	NN	O	O
stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
to	NN	O	O
induce	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
to	NN	O	O
undergo	NN	O	O
15	NN	O	O
to	NN	O	O
22	NN	O	O
cell	NN	O	O
divisions	NN	O	O
,	NN	O	O
corresponding	NN	O	O
to	NN	O	O
a	NN	O	O
10	NN	O	O
(	NN	O	O
5	NN	O	O
)	NN	O	O
-	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
-fold	NN	O	O
amplification	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Dex	NN	O	O
acted	NN	O	O
directly	NN	O	O
on	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
and	NN	O	O
maintained	NN	O	O
the	NN	O	O
colony-forming	NN	O	O
capacity	NN	O	O
of	NN	O	O
the	NN	O	O
progenitor	NN	O	B-cell_type
cells	NN	O	I-cell_type
expanded	NN	O	O
in	NN	O	O
liquid	NN	O	B-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
hormone	NN	O	O
delayed	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
into	NN	O	O
erythrocytes	NN	O	B-cell_type
,	NN	O	O
which	NN	O	O
was	NN	O	O
assayed	NN	O	O
by	NN	O	O
morphology	NN	O	O
,	NN	O	O
hemoglobin	NN	O	B-protein
accumulation	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
characteristic	NN	O	O
for	NN	O	O
immature	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Sustained	NN	O	O
proliferation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
could	NN	O	O
be	NN	O	O
induced	NN	O	O
equally	NN	O	O
well	NN	O	O
from	NN	O	O
purified	NN	O	B-cell_line
erythroid	NN	O	I-cell_line
burst-forming	NN	O	I-cell_line
units	NN	O	I-cell_line
(	NN	O	O
BFU-E	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
from	NN	O	O
CD34	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
blast	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
from	NN	O	O
bone	NN	O	O
marrow	NN	O	O
depleted	NN	O	O
from	NN	O	O
CD34	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

NF-kappaB	NN	O	B-protein
functions	NN	O	O
as	NN	O	O
both	NN	O	O
a	NN	O	O
proapoptotic	NN	O	B-protein
and	NN	O	I-protein
antiapoptotic	NN	O	I-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
within	NN	O	O
a	NN	O	O
single	NN	O	O
cell	NN	O	O
type	NN	O	O
.	NN	O	O

Recently	NN	O	O
NF-kappaB	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
have	NN	O	O
both	NN	O	O
proapoptotic	NN	O	O
and	NN	O	O
antiapoptotic	NN	O	O
functions	NN	O	O
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
,	NN	O	O
both	NN	O	O
T	NN	O	O
cell	NN	O	O
activators	NN	O	O
and	NN	O	O
glucocorticoids	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
blockade	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
IkappaBalpha	NN	O	I-protein
,	NN	O	O
has	NN	O	O
opposite	NN	O	O
effects	NN	O	O
on	NN	O	O
these	NN	O	O
two	NN	O	O
apoptotic	NN	O	O
signals	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
PMA	NN	O	O
plus	NN	O	O
ionomycin	NN	O	O
(	NN	O	O
P/I	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
Ligand	NN	O	I-protein
(	NN	O	O
FasL	NN	O	B-protein
)	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
inhibits	NN	O	O
the	NN	O	O
P/I	NN	O	O
mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
FasL	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
decreases	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
these	NN	O	O
cultures	NN	O	O
,	NN	O	O
thus	NN	O	O
establishing	NN	O	O
NF-kappaB	NN	O	B-protein
as	NN	O	O
a	NN	O	O
proapoptotic	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
this	NN	O	O
context	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
confers	NN	O	O
a	NN	O	O
tenfold	NN	O	O
increase	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	O
mediated	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
establishing	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
also	NN	O	O
functions	NN	O	O
as	NN	O	O
an	NN	O	O
antiapoptotic	NN	O	O
factor	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
is	NN	O	O
a	NN	O	O
context-dependent	NN	O	O
apoptosis	NN	O	O
regulator	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggests	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
may	NN	O	O
function	NN	O	O
as	NN	O	O
an	NN	O	O
antiapoptotic	NN	O	O
factor	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
while	NN	O	O
functioning	NN	O	O
as	NN	O	O
a	NN	O	O
proapoptotic	NN	O	O
factor	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

SOCS-3	NN	O	B-protein
is	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
interleukin-2	NN	O	B-protein
and	NN	O	O
suppresses	NN	O	O
STAT5	NN	O	B-protein
phosphorylation	NN	O	O
and	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Members	NN	O	O
of	NN	O	O
the	NN	O	O
recently	NN	O	O
discovered	NN	O	O
SOCS/CIS/SSI	NN	O	B-protein
family	NN	O	I-protein
have	NN	O	O
been	NN	O	O
proposed	NN	O	O
as	NN	O	O
regulators	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
signaling	NN	O	O
,	NN	O	O
and	NN	O	O
while	NN	O	O
targets	NN	O	O
and	NN	O	O
mechanisms	NN	O	O
have	NN	O	O
been	NN	O	O
suggested	NN	O	O
for	NN	O	O
some	NN	O	O
family	NN	O	O
members	NN	O	O
,	NN	O	O
the	NN	O	O
precise	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
defined	NN	O	O
.	NN	O	O

To	NN	O	O
date	NN	O	O
no	NN	O	O
SOCS	NN	O	B-protein
proteins	NN	O	I-protein
have	NN	O	O
been	NN	O	O
specifically	NN	O	O
implicated	NN	O	O
in	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
SOCS-3	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
both	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

SOCS-3	NN	O	B-protein
protein	NN	O	O
was	NN	O	O
detectable	NN	O	O
as	NN	O	O
early	NN	O	O
as	NN	O	O
30	NN	O	O
min	NN	O	O
following	NN	O	O
IL-2	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
while	NN	O	O
CIS	NN	O	B-protein
was	NN	O	O
seen	NN	O	O
only	NN	O	O
at	NN	O	O
low	NN	O	O
levels	NN	O	O
after	NN	O	O
2	NN	O	O
h	NN	O	O
.	NN	O	O

Unlike	NN	O	O
CIS	NN	O	B-protein
,	NN	O	O
SOCS-3	NN	O	B-protein
was	NN	O	O
rapidly	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
SOCS-3	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
upon	NN	O	O
coexpression	NN	O	O
with	NN	O	O
Jak1	NN	O	B-protein
and	NN	O	O
Jak2	NN	O	B-protein
but	NN	O	O
only	NN	O	O
weakly	NN	O	O
with	NN	O	O
Jak3	NN	O	B-protein
.	NN	O	O

In	NN	O	O
these	NN	O	O
experiments	NN	O	O
,	NN	O	O
SOCS-3	NN	O	B-protein
associated	NN	O	O
with	NN	O	O
Jak1	NN	O	B-protein
and	NN	O	O
inhibited	NN	O	O
Jak1	NN	O	B-protein
phosphorylation	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
inhibition	NN	O	O
was	NN	O	O
markedly	NN	O	O
enhanced	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
beta	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
IL-2Rbeta	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
following	NN	O	O
IL-2	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
SOCS-3	NN	O	B-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
and	NN	O	O
in	NN	O	O
particular	NN	O	O
tyrosine	NN	O	B-protein
phosphorylated	NN	O	I-protein
Jak1	NN	O	I-protein
and	NN	O	O
IL-2Rbeta	NN	O	B-protein
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
expressing	NN	O	O
SOCS-3	NN	O	B-protein
but	NN	O	O
not	NN	O	O
CIS	NN	O	B-protein
,	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT5b	NN	O	B-protein
was	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
,	NN	O	O
while	NN	O	O
there	NN	O	O
was	NN	O	O
only	NN	O	O
a	NN	O	O
weak	NN	O	O
effect	NN	O	O
on	NN	O	O
IL-3-mediated	NN	O	B-protein
STAT5b	NN	O	I-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
proliferation	NN	O	O
induced	NN	O	O
by	NN	O	O
both	NN	O	O
IL-2	NN	O	B-protein
-and	NN	O	O
IL-3	NN	O	B-protein
was	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
SOCS-3	NN	O	B-protein
.	NN	O	O

The	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
when	NN	O	O
SOCS-3	NN	O	B-protein
is	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
it	NN	O	O
acts	NN	O	O
to	NN	O	O
inhibit	NN	O	O
IL-2	NN	O	B-protein
responses	NN	O	O
in	NN	O	O
a	NN	O	O
classical	NN	O	O
negative	NN	O	O
feedback	NN	O	O
loop	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
are	NN	O	O
markedly	NN	O	O
deficient	NN	O	O
in	NN	O	O
REF-1	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Although	NN	O	O
many	NN	O	O
functions	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
are	NN	O	O
altered	NN	O	O
compared	NN	O	O
with	NN	O	O
their	NN	O	O
precursor	NN	O	B-cell_type
cell	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
blood	NN	O	B-cell_type
monocyte	NN	O	I-cell_type
(	NN	O	O
monocyte	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
the	NN	O	O
reason	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
for	NN	O	O
these	NN	O	O
functional	NN	O	O
changes	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
determined	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
reported	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
do	NN	O	O
not	NN	O	O
express	NN	O	O
AP-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
(	NN	O	O
Monick	NN	O	O
,	NN	O	O
M.	NN	O	O
M.	NN	O	O
,	NN	O	O
Carter	NN	O	O
,	NN	O	O
A.	NN	O	O
B.	NN	O	O
,	NN	O	O
Gudmundsson	NN	O	O
,	NN	O	O
G.	NN	O	O
,	NN	O	O
Geist	NN	O	O
,	NN	O	O
L.	NN	O	O
J.	NN	O	O
,	NN	O	O
and	NN	O	O
Hunninghake	NN	O	O
,	NN	O	O
G.	NN	O	O
W.	NN	O	O
(	NN	O	O
1998	NN	O	O
)	NN	O	O
Am.	NN	O	O
J.	NN	O	O
Physiol.	NN	O	O
275	NN	O	O
,	NN	O	O
L389-L397	NN	O	O
)	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
why	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
do	NN	O	O
not	NN	O	O
express	NN	O	O
AP-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
we	NN	O	O
first	NN	O	O
showed	NN	O	O
that	NN	O	O
there	NN	O	O
was	NN	O	O
not	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
FOS	NN	O	B-protein
and	NN	O	I-protein
JUN	NN	O	I-protein
proteins	NN	O	I-protein
that	NN	O	O
make	NN	O	O
up	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
complex	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
amounts	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
REF-1	NN	O	B-protein
(	NN	O	O
which	NN	O	O
regulates	NN	O	O
AP-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
altering	NN	O	O
the	NN	O	O
redox	NN	O	O
status	NN	O	O
of	NN	O	O
FOS	NN	O	B-protein
and	NN	O	O
JUN	NN	O	B-protein
proteins	NN	O	I-protein
)	NN	O	O
,	NN	O	O
in	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
compared	NN	O	O
with	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
differentiation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
to	NN	O	O
a	NN	O	O
macrophage-like	NN	O	B-cell_type
cell	NN	O	I-cell_type
resulted	NN	O	O
in	NN	O	O
decreased	NN	O	O
amounts	NN	O	O
of	NN	O	O
REF-1	NN	O	B-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
REF-1	NN	O	B-protein
from	NN	O	O
activated	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
to	NN	O	O
alveolar	NN	O	O
macrophage	NN	O	O
nuclear	NN	O	O
proteins	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
marked	NN	O	O
increase	NN	O	O
in	NN	O	O
AP-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
differentiation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
into	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
loss	NN	O	O
of	NN	O	O
REF-1	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
may	NN	O	O
explain	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
differences	NN	O	O
observed	NN	O	O
for	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
compared	NN	O	O
with	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Development	NN	O	O
and	NN	O	O
maturation	NN	O	O
of	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
.	NN	O	O

The	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
are	NN	O	O
located	NN	O	O
at	NN	O	O
strategic	NN	O	O
sites	NN	O	O
where	NN	O	O
foreign	NN	O	B-protein
antigens	NN	O	I-protein
can	NN	O	O
be	NN	O	O
efficiently	NN	O	O
brought	NN	O	O
together	NN	O	O
with	NN	O	O
immune	NN	O	B-cell_type
system	NN	O	I-cell_type
regulatory	NN	O	I-cell_type
and	NN	O	I-cell_type
effector	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
organized	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
enhance	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
antigen	NN	O	O
recognition	NN	O	O
and	NN	O	O
to	NN	O	O
support	NN	O	O
proper	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
activation	NN	O	O
and	NN	O	O
maturation	NN	O	O
of	NN	O	O
the	NN	O	O
antigen-responsive	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
a	NN	O	O
substantial	NN	O	O
amount	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
cellular	NN	O	B-DNA
elements	NN	O	I-DNA
that	NN	O	O
compose	NN	O	O
the	NN	O	O
lymphoid	NN	O	O
and	NN	O	O
nonlymphoid	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
,	NN	O	O
information	NN	O	O
concerning	NN	O	O
the	NN	O	O
signals	NN	O	O
that	NN	O	O
control	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
tissues	NN	O	O
and	NN	O	O
that	NN	O	O
maintain	NN	O	O
the	NN	O	O
organized	NN	O	O
tissue	NN	O	O
microenvironment	NN	O	O
remain	NN	O	O
undefined	NN	O	O
.	NN	O	O

Studies	NN	O	O
over	NN	O	O
the	NN	O	O
past	NN	O	O
few	NN	O	O
years	NN	O	O
have	NN	O	O
identified	NN	O	O
lymphotoxin	NN	O	O
as	NN	O	O
a	NN	O	O
critical	NN	O	O
signaling	NN	O	O
molecule	NN	O	O
not	NN	O	O
only	NN	O	O
for	NN	O	O
the	NN	O	O
organogenesis	NN	O	O
of	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
but	NN	O	O
for	NN	O	O
the	NN	O	O
maintenance	NN	O	O
of	NN	O	O
aspects	NN	O	O
of	NN	O	O
their	NN	O	O
microarchitecture	NN	O	O
as	NN	O	O
well	NN	O	O
.	NN	O	O

Additional	NN	O	O
signaling	NN	O	O
molecules	NN	O	O
that	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
normal	NN	O	O
lymphoid	NN	O	O
tissue	NN	O	O
structure	NN	O	O
are	NN	O	O
being	NN	O	O
identified	NN	O	O
at	NN	O	O
an	NN	O	O
accelerating	NN	O	O
pace	NN	O	O
.	NN	O	O

Analyses	NN	O	O
of	NN	O	O
mouse	NN	O	O
strains	NN	O	O
with	NN	O	O
congenital	NN	O	O
defects	NN	O	O
in	NN	O	O
different	NN	O	O
aspects	NN	O	O
of	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
tissue	NN	O	O
development	NN	O	O
are	NN	O	O
beginning	NN	O	O
to	NN	O	O
clarify	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
tissues	NN	O	O
in	NN	O	O
immune	NN	O	O
responses	NN	O	O
and	NN	O	O
host	NN	O	O
defense	NN	O	O
.	NN	O	O

This	NN	O	O
review	NN	O	O
focuses	NN	O	O
on	NN	O	O
studies	NN	O	O
defining	NN	O	O
recently	NN	O	O
identified	NN	O	O
crucial	NN	O	O
signals	NN	O	O
for	NN	O	O
the	NN	O	O
biogenesis	NN	O	O
of	NN	O	O
secondary	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
and	NN	O	O
for	NN	O	O
the	NN	O	O
maintenance	NN	O	O
of	NN	O	O
their	NN	O	O
proper	NN	O	O
microarchitecture	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
discusses	NN	O	O
new	NN	O	O
insights	NN	O	O
into	NN	O	O
how	NN	O	O
the	NN	O	O
structure	NN	O	O
of	NN	O	O
these	NN	O	O
tissues	NN	O	O
supports	NN	O	O
effective	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Modulation	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
expression	NN	O	O
:	NN	O	O
distinct	NN	O	O
regulatory	NN	O	O
pathways	NN	O	O
during	NN	O	O
activation	NN	O	O
and	NN	O	O
replicative	NN	O	O
senescence	NN	O	O
.	NN	O	O

The	NN	O	O
costimulatory	NN	O	B-protein
molecule	NN	O	I-protein
CD28	NN	O	B-protein
has	NN	O	O
a	NN	O	O
restricted	NN	O	O
tissue	NN	O	O
distribution	NN	O	O
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
some	NN	O	O
plasmacytoma	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
CD28	NN	O	B-protein
is	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
,	NN	O	O
its	NN	O	O
expression	NN	O	O
is	NN	O	O
transiently	NN	O	O
down-regulated	NN	O	O
following	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
declines	NN	O	O
progressively	NN	O	O
with	NN	O	O
in	NN	O	O
vitro	NN	O	O
senescence	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
,	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
,	NN	O	O
less	NN	O	O
frequently	NN	O	O
,	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
completely	NN	O	O
lose	NN	O	O
CD28	NN	O	B-protein
surface	NN	O	O
expression	NN	O	O
during	NN	O	O
chronic	NN	O	O
infections	NN	O	O
and	NN	O	O
with	NN	O	O
aging	NN	O	O
.	NN	O	O

This	NN	O	O
correlates	NN	O	O
with	NN	O	O
changes	NN	O	O
of	NN	O	O
nuclear	NN	O	O
protein-binding	NN	O	O
activities	NN	O	O
to	NN	O	O
two	NN	O	O
motifs	NN	O	O
,	NN	O	O
site	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
beta	NN	O	I-protein
,	NN	O	O
within	NN	O	O
the	NN	O	O
CD28	NN	O	B-DNA
minimal	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
alpha-	NN	O	O
and	NN	O	O
beta-bound	NN	O	O
complexes	NN	O	O
are	NN	O	O
found	NN	O	O
only	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
,	NN	O	O
in	NN	O	O
CD28+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
in	NN	O	O
some	NN	O	O
transformed	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
complexes	NN	O	O
are	NN	O	O
coordinately	NN	O	O
expressed	NN	O	O
except	NN	O	O
during	NN	O	O
replicative	NN	O	O
senescence	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
down-modulation	NN	O	O
of	NN	O	O
site	NN	O	B-protein
beta	NN	O	I-protein
-but	NN	O	O
not	NN	O	O
site	NN	O	B-protein
alpha	NN	O	I-protein
-binding	NN	O	O
activities	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
induces	NN	O	O
a	NN	O	O
parallel	NN	O	O
decline	NN	O	O
in	NN	O	O
both	NN	O	O
site	NN	O	O
alpha-	NN	O	O
and	NN	O	O
beta-binding	NN	O	O
activities	NN	O	O
.	NN	O	O

CD4+	NN	O	O
and	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
differ	NN	O	O
in	NN	O	O
their	NN	O	O
beta-binding	NN	O	O
profiles	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
explain	NN	O	O
the	NN	O	O
more	NN	O	O
pronounced	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
in	NN	O	O
senescent	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
expanded	NN	O	O
CD4+CD28null	NN	O	B-cell_line
and	NN	O	I-cell_line
CD8+CD28null	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
uniformly	NN	O	O
lack	NN	O	O
alpha-	NN	O	B-protein
and	NN	O	I-protein
beta-	NN	O	I-protein
bound	NN	O	I-protein
complexes	NN	O	I-protein
,	NN	O	O
resembling	NN	O	O
the	NN	O	O
pattern	NN	O	O
seen	NN	O	O
in	NN	O	O
chronically	NN	O	B-cell_type
activated	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
not	NN	O	O
of	NN	O	O
senescent	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
role	NN	O	O
for	NN	O	O
RanBP1	NN	O	B-protein
in	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
CRM1	NN	O	B-protein
from	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
pore	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
a	NN	O	O
terminal	NN	O	O
step	NN	O	O
of	NN	O	O
nuclear	NN	O	O
export	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
developed	NN	O	O
an	NN	O	O
assay	NN	O	O
in	NN	O	O
which	NN	O	O
nuclear	NN	O	O
export	NN	O	O
of	NN	O	O
the	NN	O	O
shuttling	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NFAT	NN	O	I-protein
(	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
)	NN	O	O
can	NN	O	O
be	NN	O	O
reconstituted	NN	O	O
in	NN	O	O
permeabilized	NN	O	O
cells	NN	O	O
with	NN	O	O
the	NN	O	O
GTPase	NN	O	B-protein
Ran	NN	O	B-protein
and	NN	O	O
the	NN	O	O
nuclear	NN	O	O
export	NN	O	O
receptor	NN	O	O
CRM1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
used	NN	O	O
this	NN	O	O
assay	NN	O	O
to	NN	O	O
identify	NN	O	O
another	NN	O	O
export	NN	O	O
factor	NN	O	O
.	NN	O	O

After	NN	O	O
preincubation	NN	O	O
of	NN	O	O
permeabilized	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
a	NN	O	O
Ran	NN	O	B-protein
mutant	NN	O	I-protein
that	NN	O	O
can	NN	O	O
not	NN	O	O
hydrolyze	NN	O	O
GTP	NN	O	O
(	NN	O	O
RanQ69L	NN	O	B-protein
)	NN	O	O
,	NN	O	O
cytosol	NN	O	O
supports	NN	O	O
NFAT	NN	O	B-protein
export	NN	O	O
,	NN	O	O
but	NN	O	O
CRM1	NN	O	B-protein
and	NN	O	O
Ran	NN	O	B-protein
alone	NN	O	O
do	NN	O	O
not	NN	O	O
.	NN	O	O

The	NN	O	O
RanQ69L	NN	O	B-protein
preincubation	NN	O	O
leads	NN	O	O
to	NN	O	O
accumulation	NN	O	O
of	NN	O	O
CRM1	NN	O	B-protein
at	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
periphery	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
pore	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	O
NPC	NN	O	B-protein
)	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
the	NN	O	O
p62	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
Can/Nup214	NN	O	O
.	NN	O	O

Ran	NN	O	B-protein
GTP-dependent	NN	O	O
association	NN	O	O
of	NN	O	O
CRM1	NN	O	B-protein
with	NN	O	O
these	NN	O	O
nucleoporins	NN	O	B-protein
was	NN	O	O
reconstituted	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

By	NN	O	O
biochemical	NN	O	O
fractionation	NN	O	O
and	NN	O	O
reconstitution	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
RanBP1	NN	O	B-protein
restores	NN	O	O
nuclear	NN	O	O
export	NN	O	O
after	NN	O	O
the	NN	O	O
RanQ69L	NN	O	B-protein
preincubation	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
stimulates	NN	O	O
nuclear	NN	O	O
export	NN	O	O
in	NN	O	O
cells	NN	O	O
that	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
preincubated	NN	O	O
with	NN	O	O
RanQ69L	NN	O	B-protein
.	NN	O	O

RanBP1	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
Ran-binding	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
nucleoporin	NN	O	I-protein
RanBP2	NN	O	B-protein
promote	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
CRM1	NN	O	B-protein
from	NN	O	O
the	NN	O	O
NPC	NN	O	B-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
Ran	NN	O	B-protein
GTP	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
targeting	NN	O	O
of	NN	O	O
export	NN	O	O
complexes	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
side	NN	O	O
of	NN	O	O
the	NN	O	O
NPC	NN	O	B-protein
and	NN	O	O
that	NN	O	O
RanBP1	NN	O	B-protein
and	NN	O	O
probably	NN	O	O
RanBP2	NN	O	B-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
dissociation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
export	NN	O	I-protein
complexes	NN	O	I-protein
from	NN	O	O
the	NN	O	O
NPC	NN	O	B-protein
in	NN	O	O
a	NN	O	O
terminal	NN	O	O
step	NN	O	O
of	NN	O	O
transport	NN	O	O
.	NN	O	O

-DOCSTART-	O

c-Myc	NN	O	B-protein
and	NN	O	O
E1A	NN	O	B-protein
induced	NN	O	O
cellular	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
activated	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
involves	NN	O	O
cytotoxic	NN	O	O
granules	NN	O	O
as	NN	O	O
death	NN	O	O
effectors	NN	O	O
.	NN	O	O

The	NN	O	O
contact	NN	O	O
of	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
foreign	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
with	NN	O	O
certain	NN	O	O
virus-infected	NN	O	B-cell_type
or	NN	O	I-cell_type
tumor	NN	O	I-cell_type
cells	NN	O	I-cell_type
triggers	NN	O	O
the	NN	O	O
cytolytic	NN	O	O
machinery	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
triggering	NN	O	O
leads	NN	O	O
to	NN	O	O
exocytosis	NN	O	O
of	NN	O	O
the	NN	O	O
cytotoxic	NN	O	B-cell_type
NK	NN	O	I-cell_type
cell	NN	O	I-cell_type
granules	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
oncoproteins	NN	O	B-protein
c-Myc	NN	O	B-protein
and	NN	O	O
E1A	NN	O	B-protein
render	NN	O	O
cells	NN	O	O
vulnerable	NN	O	O
to	NN	O	O
NK	NN	O	O
cell	NN	O	O
mediated	NN	O	O
cytolysis	NN	O	O
yet	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
sensitization	NN	O	O
are	NN	O	O
not	NN	O	O
well	NN	O	O
understood	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
model	NN	O	O
where	NN	O	O
foreign	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
rat	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
)	NN	O	O
were	NN	O	O
cocultured	NN	O	O
with	NN	O	O
human	NN	O	B-cell_type
IL-2	NN	O	I-cell_type
activated	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
capable	NN	O	O
of	NN	O	O
efficiently	NN	O	O
killing	NN	O	O
their	NN	O	O
targets	NN	O	O
only	NN	O	O
if	NN	O	O
the	NN	O	O
cells	NN	O	O
overexpressed	NN	O	O
the	NN	O	O
oncogene	NN	O	O
c-Myc	NN	O	B-protein
or	NN	O	O
E1A	NN	O	B-protein
.	NN	O	O

Both	NN	O	O
the	NN	O	O
parental	NN	O	O
and	NN	O	O
the	NN	O	O
oncogene	NN	O	B-cell_type
expressing	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
similarly	NN	O	O
triggered	NN	O	O
phosphoinositide	NN	O	O
hydrolysis	NN	O	O
in	NN	O	O
the	NN	O	O
bound	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
cytolytically	NN	O	O
activated	NN	O	O
in	NN	O	O
contact	NN	O	O
with	NN	O	O
both	NN	O	O
resistant	NN	O	O
parental	NN	O	O
and	NN	O	O
oncogene	NN	O	B-cell_type
expressing	NN	O	I-cell_type
sensitive	NN	O	I-cell_type
target	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
cell	NN	O	O
death	NN	O	O
was	NN	O	O
independent	NN	O	O
of	NN	O	O
wild-type	NN	O	B-protein
p53	NN	O	I-protein
and	NN	O	O
was	NN	O	O
not	NN	O	O
inhibited	NN	O	O
by	NN	O	O
an	NN	O	O
anti-apoptotic	NN	O	B-protein
protein	NN	O	I-protein
EIB19K	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
provided	NN	O	O
evidence	NN	O	O
that	NN	O	O
c-Myc	NN	O	B-protein
and	NN	O	O
E1A	NN	O	B-protein
activated	NN	O	O
the	NN	O	O
NK	NN	O	O
cell	NN	O	O
induced	NN	O	O
cytolysis	NN	O	O
at	NN	O	O
a	NN	O	O
post-triggering	NN	O	O
stage	NN	O	O
of	NN	O	O
NK	NN	O	O
cell-target	NN	O	O
cell	NN	O	O
interaction	NN	O	O
.	NN	O	O

In	NN	O	O
consistence	NN	O	O
,	NN	O	O
the	NN	O	O
c-Myc	NN	O	B-protein
and	NN	O	O
E1A	NN	O	B-protein
overexpressing	NN	O	O
fibroblasts	NN	O	B-cell_type
were	NN	O	O
more	NN	O	O
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
cytolytic	NN	O	O
effects	NN	O	O
of	NN	O	O
isolated	NN	O	O
NK	NN	O	O
cell-derived	NN	O	O
granules	NN	O	O
than	NN	O	O
parental	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
oncogenes	NN	O	B-DNA
activate	NN	O	O
the	NN	O	O
cytotoxicity	NN	O	O
of	NN	O	O
NK	NN	O	O
cell	NN	O	O
granules	NN	O	O
.	NN	O	O

This	NN	O	O
mechanism	NN	O	O
can	NN	O	O
have	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
directing	NN	O	O
the	NN	O	O
cytolytic	NN	O	O
action	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
towards	NN	O	O
the	NN	O	O
virus-infected	NN	O	B-cell_type
and	NN	O	I-cell_type
cancer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Unicellular-unilineage	NN	O	B-cell_line
erythropoietic	NN	O	I-cell_line
cultures	NN	O	I-cell_line
:	NN	O	O
molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
regulatory	NN	O	O
gene	NN	O	O
expression	NN	O	O
at	NN	O	O
sibling	NN	O	O
cell	NN	O	O
level	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
studies	NN	O	O
on	NN	O	O
hematopoietic	NN	O	O
control	NN	O	O
mechanisms	NN	O	O
have	NN	O	O
been	NN	O	O
hampered	NN	O	O
by	NN	O	O
the	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
the	NN	O	O
analyzed	NN	O	O
cell	NN	O	O
populations	NN	O	O
,	NN	O	O
ie	NN	O	O
,	NN	O	O
lack	NN	O	O
of	NN	O	O
lineage	NN	O	O
specificity	NN	O	O
and	NN	O	O
developmental	NN	O	O
stage	NN	O	O
homogeneity	NN	O	O
of	NN	O	O
progenitor/precursor	NN	O	B-cell_type
cells	NN	O	I-cell_type
growing	NN	O	O
in	NN	O	O
culture	NN	O	O
.	NN	O	O

We	NN	O	O
developed	NN	O	O
unicellular	NN	O	O
culture	NN	O	O
systems	NN	O	O
for	NN	O	O
unilineage	NN	O	O
differentiation	NN	O	O
of	NN	O	O
purified	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
followed	NN	O	O
by	NN	O	O
daughter	NN	O	O
cell	NN	O	O
analysis	NN	O	O
at	NN	O	O
cellular	NN	O	O
and	NN	O	O
molecular	NN	O	O
level	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
culture	NN	O	O
system	NN	O	O
reported	NN	O	O
here	NN	O	O
,	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
growth	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
GF	NN	O	B-protein
)	NN	O	O
stimulus	NN	O	O
induces	NN	O	O
cord	NN	O	B-cell_type
blood	NN	O	I-cell_type
(	NN	O	I-cell_type
CB	NN	O	I-cell_type
)	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
proliferate	NN	O	O
and	NN	O	O
differentiate/mature	NN	O	O
exclusively	NN	O	O
along	NN	O	O
the	NN	O	O
erythroid	NN	O	B-cell_type
lineage	NN	O	I-cell_type
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
this	NN	O	O
erythropoietic	NN	O	O
wave	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
less	NN	O	O
than	NN	O	O
4	NN	O	O
%	NN	O	O
apoptotic	NN	O	O
cells	NN	O	O
;	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
asymmetric	NN	O	O
divisions	NN	O	O
are	NN	O	O
virtually	NN	O	O
absent	NN	O	O
,	NN	O	O
ie	NN	O	O
,	NN	O	O
nonresponsive	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
with	NN	O	O
no	NN	O	O
erythropoietic	NN	O	O
potential	NN	O	O
are	NN	O	O
forced	NN	O	O
into	NN	O	O
apoptosis	NN	O	O
;	NN	O	O
(	NN	O	O
4	NN	O	O
)	NN	O	O
the	NN	O	O
system	NN	O	O
is	NN	O	O
cell	NN	O	O
division	NN	O	O
controlled	NN	O	O
(	NN	O	O
cdc	NN	O	O
)	NN	O	O
,	NN	O	O
ie	NN	O	O
,	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
divisions	NN	O	O
performed	NN	O	O
by	NN	O	O
each	NN	O	O
cell	NN	O	O
is	NN	O	O
monitored	NN	O	O
.	NN	O	O

Single-cell	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
analysis	NN	O	O
was	NN	O	O
applied	NN	O	O
to	NN	O	O
this	NN	O	O
culture	NN	O	O
system	NN	O	O
to	NN	O	O
investigate	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
diverse	NN	O	O
receptors	NN	O	B-protein
,	NN	O	O
markers	NN	O	B-protein
of	NN	O	I-protein
differentiation	NN	O	I-protein
,	NN	O	O
and	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
EKLF	NN	O	B-protein
,	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
GATA-2	NN	O	B-protein
,	NN	O	O
p45	NN	O	B-protein
NF-E2	NN	O	I-protein
,	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
SCL/Tal1	NN	O	B-protein
)	NN	O	O
at	NN	O	O
discrete	NN	O	O
stages	NN	O	O
of	NN	O	O
erythropoietic	NN	O	O
development	NN	O	O
.	NN	O	O

Freshly	NN	O	O
isolated	NN	O	O
CD34	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
expressed	NN	O	O
CD34	NN	O	B-protein
,	NN	O	O
c-kit	NN	O	B-protein
,	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GATA-2	NN	O	B-protein
but	NN	O	O
did	NN	O	O
not	NN	O	O
express	NN	O	O
CD36	NN	O	B-protein
,	NN	O	O
erythropoietin	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
EpoR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
SCL/Tal1	NN	O	B-protein
,	NN	O	O
EKLF	NN	O	B-protein
,	NN	O	O
NF-E2	NN	O	O
,	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
or	NN	O	O
glyocophorin	NN	O	B-protein
A	NN	O	I-protein
(	NN	O	O
GPA	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
early	NN	O	O
to	NN	O	O
intermediate	NN	O	O
stages	NN	O	O
of	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
we	NN	O	O
monitored	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
CD36	NN	O	B-protein
,	NN	O	O
Tal1	NN	O	B-protein
,	NN	O	O
EKLF	NN	O	B-protein
,	NN	O	O
NF-E2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GATA-1	NN	O	B-protein
that	NN	O	O
preceeded	NN	O	O
expression	NN	O	O
of	NN	O	O
EpoR	NN	O	B-protein
.	NN	O	O

In	NN	O	O
late	NN	O	O
stages	NN	O	O
of	NN	O	O
erythroid	NN	O	O
maturation	NN	O	O
,	NN	O	O
GPA	NN	O	B-protein
was	NN	O	O
upregulated	NN	O	O
,	NN	O	O
whereas	NN	O	O
CD34	NN	O	B-protein
,	NN	O	O
c-kit	NN	O	B-protein
,	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GATA-2	NN	O	B-protein
were	NN	O	O
barely	NN	O	O
or	NN	O	O
not	NN	O	O
detected	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
competitive	NN	O	O
single-cell	NN	O	O
RT-PCR	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
assay	NN	O	O
CD34	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
sibling	NN	O	B-cell_line
CD34	NN	O	I-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
CD38	NN	O	I-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
differentiating	NN	O	O
in	NN	O	O
unilineage	NN	O	B-cell_line
erythroid	NN	O	I-cell_line
cultures	NN	O	I-cell_line
:	NN	O	O
this	NN	O	O
analysis	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
semiquantitate	NN	O	O
the	NN	O	O
gradual	NN	O	O
downmodulation	NN	O	O
of	NN	O	O
CD34	NN	O	B-RNA
mRNA	NN	O	I-RNA
from	NN	O	O
progenitor	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
their	NN	O	O
differentiating	NN	O	B-cell_line
erythroid	NN	O	I-cell_line
progeny	NN	O	I-cell_line
.	NN	O	O

It	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
this	NN	O	O
novel	NN	O	O
culture	NN	O	O
system	NN	O	O
,	NN	O	O
coupled	NN	O	O
with	NN	O	O
single-cell	NN	O	O
RT-PCR	NN	O	O
analysis	NN	O	O
,	NN	O	O
may	NN	O	O
eliminate	NN	O	O
the	NN	O	O
ambiguities	NN	O	O
intrinsic	NN	O	O
to	NN	O	O
molecular	NN	O	O
studies	NN	O	O
on	NN	O	O
heterogeneous	NN	O	O
populations	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitors/precursors	NN	O	I-cell_type
growing	NN	O	O
in	NN	O	O
culture	NN	O	O
,	NN	O	O
particularly	NN	O	O
in	NN	O	O
the	NN	O	O
initial	NN	O	O
stages	NN	O	O
of	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differences	NN	O	O
in	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-protein
associated	NN	O	I-protein
JAK/STAT	NN	O	I-protein
proteins	NN	O	I-protein
between	NN	O	O
HTLV-I	NN	O	B-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
,	NN	O	I-cell_line
IL-2-independent	NN	O	I-cell_line
and	NN	O	I-cell_line
IL-2-dependent	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
uncultured	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
lymphoma/leukemia	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
activation	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-protein
-associated	NN	O	O
(	NN	O	O
Jak/STAT	NN	O	B-protein
)	NN	O	O
pathway	NN	O	O
in	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-cell_line
infected	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
examined	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak3	NN	O	B-protein
,	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT5	NN	O	B-protein
in	NN	O	O
several	NN	O	O
HTLV-I	NN	O	B-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
uncultured	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
lymphoma/leukemia	NN	O	O
(	NN	O	O
ATLL	NN	O	O
)	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
basal	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak3	NN	O	B-protein
and	NN	O	O
,	NN	O	O
usually	NN	O	O
,	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
all	NN	O	O
four	NN	O	O
IL-2-independent	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
tested	NN	O	O
,	NN	O	O
but	NN	O	O
in	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
IL-2-dependent	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
detectable	NN	O	O
basal	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak3	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
in	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
ATLL	NN	O	O
patients	NN	O	O
(	NN	O	O
0/8	NN	O	O
and	NN	O	O
0/3	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
stimulation	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
Jak3	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
phosphorylation	NN	O	O
in	NN	O	O
both	NN	O	O
leukemic	NN	O	B-cell_type
ATLL	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
IL-2-dependent	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
SHP-1	NN	O	B-protein
phosphatase	NN	O	I-protein
which	NN	O	O
is	NN	O	O
a	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
signaling	NN	O	O
,	NN	O	O
was	NN	O	O
lost	NN	O	O
in	NN	O	O
most	NN	O	O
IL-2	NN	O	B-cell_line
independent	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
3/4	NN	O	O
)	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
leukemic	NN	O	O
ATLL	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
0/3	NN	O	O
)	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
313	NN	O	O
)	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
control	NN	O	O
,	NN	O	O
HTLV-I	NN	O	B-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
resistant	NN	O	O
to	NN	O	O
rapamycin	NN	O	O
and	NN	O	O
its	NN	O	O
novel	NN	O	O
analog	NN	O	O
RAD	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
HTLV-I	NN	O	O
infection	NN	O	O
per	NN	O	O
se	NN	O	O
does	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
constitutive	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
Jak3	NN	O	B-protein
,	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT5	NN	O	B-protein
proteins	NN	O	O
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
malignant	NN	O	O
transformation	NN	O	O
in	NN	O	O
at	NN	O	O
least	NN	O	O
some	NN	O	O
cases	NN	O	O
of	NN	O	O
ATLL	NN	O	O
does	NN	O	O
not	NN	O	O
require	NN	O	O
the	NN	O	O
constitutive	NN	O	O
,	NN	O	O
but	NN	O	O
may	NN	O	O
require	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-protein
Jak/STAT	NN	O	B-protein
pathway	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
there	NN	O	O
are	NN	O	O
major	NN	O	O
differences	NN	O	O
in	NN	O	O
T-cell	NN	O	O
immortalization	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
which	NN	O	O
appear	NN	O	O
to	NN	O	O
involve	NN	O	O
SHP-1	NN	O	B-protein
and	NN	O	O
target	NN	O	O
molecules	NN	O	O
for	NN	O	O
rapamycin	NN	O	O
and	NN	O	O
RAD	NN	O	O
.	NN	O	O

-DOCSTART-	O

Clonality	NN	O	O
analysis	NN	O	O
using	NN	O	O
X-chromosome	NN	O	B-DNA
inactivation	NN	O	O
at	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
androgen	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
Humara	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Evaluation	NN	O	O
of	NN	O	O
large	NN	O	O
cohorts	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
myeloproliferative	NN	O	O
diseases	NN	O	O
,	NN	O	O
secondary	NN	O	O
neutrophilia	NN	O	O
,	NN	O	O
and	NN	O	O
reactive	NN	O	O
thrombocytosis	NN	O	O
.	NN	O	O

Chronic	NN	O	O
myeloproliferative	NN	O	O
diseases	NN	O	O
(	NN	O	O
MPDs	NN	O	O
)	NN	O	O
are	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
consistent	NN	O	O
cytogenetic	NN	O	O
or	NN	O	O
molecular	NN	O	O
abnormalities	NN	O	O
.	NN	O	O

Demonstration	NN	O	O
of	NN	O	O
clonal	NN	O	O
cell	NN	O	O
growth	NN	O	O
by	NN	O	O
analysis	NN	O	O
of	NN	O	O
X-chromosome	NN	O	B-DNA
inactivation	NN	O	O
(	NN	O	O
XCI	NN	O	O
)	NN	O	O
patterns	NN	O	O
in	NN	O	O
females	NN	O	O
provides	NN	O	O
a	NN	O	O
promising	NN	O	O
tool	NN	O	O
for	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
technique	NN	O	O
can	NN	O	O
be	NN	O	O
complicated	NN	O	O
by	NN	O	O
excessive	NN	O	O
lyonization	NN	O	O
of	NN	O	O
normal	NN	O	O
cells	NN	O	O
mimicking	NN	O	O
clonal	NN	O	O
cell	NN	O	O
growth	NN	O	O
:	NN	O	O
We	NN	O	O
analyzed	NN	O	O
XCI	NN	O	O
patterns	NN	O	O
at	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
androgen	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
HUMARA	NN	O	I-DNA
)	NN	O	I-DNA
locus	NN	O	I-DNA
in	NN	O	O
146	NN	O	O
healthy	NN	O	O
females	NN	O	O
,	NN	O	O
65	NN	O	O
women	NN	O	O
with	NN	O	O
secondary	NN	O	O
neutrophilia	NN	O	O
,	NN	O	O
31	NN	O	O
women	NN	O	O
with	NN	O	O
reactive	NN	O	O
thrombocytosis	NN	O	O
,	NN	O	O
and	NN	O	O
86	NN	O	O
women	NN	O	O
with	NN	O	O
chronic	NN	O	O
MPDs	NN	O	O
.	NN	O	O

A	NN	O	O
skewed	NN	O	O
XCI	NN	O	O
pattern	NN	O	O
with	NN	O	O
greater	NN	O	O
than	NN	O	O
75	NN	O	O
%	NN	O	O
amplification	NN	O	O
of	NN	O	O
1	NN	O	O
allele	NN	O	B-DNA
(	NN	O	O
allele	NN	O	B-DNA
ratio	NN	O	O
>	NN	O	O
3	NN	O	O
:	NN	O	O
1	NN	O	O
)	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
22	NN	O	O
(	NN	O	O
9.1	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
242	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
incidence	NN	O	O
of	NN	O	O
skewing	NN	O	O
was	NN	O	O
statistically	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
women	NN	O	O
younger	NN	O	O
than	NN	O	O
30	NN	O	O
years	NN	O	O
(	NN	O	O
2/73	NN	O	O
)	NN	O	O
compared	NN	O	O
with	NN	O	O
women	NN	O	O
older	NN	O	O
than	NN	O	O
60	NN	O	O
years	NN	O	O
(	NN	O	O
10/53	NN	O	O
)	NN	O	O
.	NN	O	O

Of	NN	O	O
86	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
chronic	NN	O	O
MPD	NN	O	O
,	NN	O	O
71	NN	O	O
(	NN	O	O
82	NN	O	O
%	NN	O	O
)	NN	O	O
exhibited	NN	O	O
an	NN	O	O
allele	NN	O	B-DNA
ratio	NN	O	O
greater	NN	O	O
than	NN	O	O
3	NN	O	O
:	NN	O	O
1	NN	O	O
,	NN	O	O
whereas	NN	O	O
only	NN	O	O
10	NN	O	O
(	NN	O	O
12	NN	O	O
%	NN	O	O
)	NN	O	O
of	NN	O	O
86	NN	O	O
age-matched	NN	O	O
control	NN	O	O
subjects	NN	O	O
showed	NN	O	O
a	NN	O	O
skewed	NN	O	O
XCI	NN	O	O
pattern	NN	O	O
.	NN	O	O

Although	NN	O	O
statistical	NN	O	O
evaluation	NN	O	O
of	NN	O	O
the	NN	O	O
data	NN	O	O
showed	NN	O	O
a	NN	O	O
significant	NN	O	O
difference	NN	O	O
between	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
chronic	NN	O	O
MPD	NN	O	O
and	NN	O	O
control	NN	O	O
subjects	NN	O	O
,	NN	O	O
proof	NN	O	O
of	NN	O	O
clonality	NN	O	O
in	NN	O	O
individual	NN	O	O
,	NN	O	O
especially	NN	O	O
elderly	NN	O	O
,	NN	O	O
patients	NN	O	O
is	NN	O	O
difficult	NN	O	O
.	NN	O	O

-DOCSTART-	O

GATA-1	NN	O	B-protein
and	NN	O	O
erythropoietin	NN	O	B-protein
cooperate	NN	O	O
to	NN	O	O
promote	NN	O	O
erythroid	NN	O	O
cell	NN	O	O
survival	NN	O	O
by	NN	O	O
regulating	NN	O	O
bcl-xL	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
normal	NN	O	O
erythropoiesis	NN	O	O
.	NN	O	O

By	NN	O	O
examining	NN	O	O
in	NN	O	O
vitro-differentiated	NN	O	B-cell_line
embryonic	NN	O	I-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
showed	NN	O	O
previously	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
committed	NN	O	B-cell_line
erythroid	NN	O	I-cell_line
precursors	NN	O	I-cell_line
fail	NN	O	O
to	NN	O	O
complete	NN	O	O
maturation	NN	O	O
and	NN	O	O
instead	NN	O	O
undergo	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
GATA-1	NN	O	B-protein
controls	NN	O	O
cell	NN	O	O
survival	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
GATA-1	NN	O	B-protein
strongly	NN	O	O
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
anti-apoptotic	NN	O	B-protein
protein	NN	O	I-protein
bcl-xL	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
related	NN	O	O
proteins	NN	O	O
bcl-2	NN	O	B-protein
and	NN	O	O
mcl-1	NN	O	B-protein
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
bcl-xL	NN	O	B-protein
in	NN	O	O
mediating	NN	O	O
GATA-1	NN	O	B-protein
-induced	NN	O	O
erythroid	NN	O	O
cell	NN	O	O
survival	NN	O	O
,	NN	O	O
in	NN	O	O
vitro-differentiated	NN	O	B-cell_line
bcl-xL-/-	NN	O	I-cell_line
embryonic	NN	O	I-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
fail	NN	O	O
to	NN	O	O
generate	NN	O	O
viable	NN	O	O
mature	NN	O	O
definitive	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
phenotype	NN	O	O
resembling	NN	O	O
that	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-DNA
gene	NN	O	I-DNA
disruption	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
erythropoietin	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
also	NN	O	O
required	NN	O	O
for	NN	O	O
erythroid	NN	O	O
cell	NN	O	O
survival	NN	O	O
,	NN	O	O
cooperates	NN	O	O
with	NN	O	O
GATA-1	NN	O	B-protein
to	NN	O	O
stimulate	NN	O	O
bcl-xL	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
to	NN	O	O
maintain	NN	O	O
erythroid	NN	O	O
cell	NN	O	O
viability	NN	O	O
during	NN	O	O
terminal	NN	O	O
maturation	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
bcl-xL	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
normal	NN	O	O
erythroid	NN	O	O
development	NN	O	O
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
regulatory	NN	O	O
hierarchy	NN	O	O
in	NN	O	O
which	NN	O	O
bcl-xL	NN	O	B-protein
is	NN	O	O
a	NN	O	O
critical	NN	O	O
downstream	NN	O	O
effector	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
erythropoietin	NN	O	B-protein
-mediated	NN	O	O
signals	NN	O	O
.	NN	O	O

-DOCSTART-	O

p70	NN	O	B-protein
(	NN	O	I-protein
s6k	NN	O	I-protein
)	NN	O	I-protein
integrates	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
and	NN	O	O
rapamycin-regulated	NN	O	O
signals	NN	O	O
for	NN	O	O
E2F	NN	O	B-protein
regulation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-protein
cytokine	NN	O	I-protein
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
uses	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
(	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
)	NN	O	O
-induced	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
to	NN	O	O
regulate	NN	O	O
E2F	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
,	NN	O	O
a	NN	O	O
critical	NN	O	O
cell	NN	O	O
cycle	NN	O	O
checkpoint	NN	O	O
.	NN	O	O

PI	NN	O	B-protein
3-kinase	NN	O	I-protein
also	NN	O	O
regulates	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
p70	NN	O	B-protein
(	NN	O	I-protein
s6k	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
the	NN	O	O
40S	NN	O	B-protein
ribosomal	NN	O	I-protein
protein	NN	O	I-protein
S6	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
a	NN	O	O
response	NN	O	O
that	NN	O	O
is	NN	O	O
abrogated	NN	O	O
by	NN	O	O
the	NN	O	O
macrolide	NN	O	O
rapamycin	NN	O	O
.	NN	O	O

This	NN	O	O
immunosuppressive	NN	O	O
drug	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
prevent	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
precise	NN	O	O
point	NN	O	O
at	NN	O	O
which	NN	O	O
rapamycin	NN	O	O
regulates	NN	O	O
T-cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
has	NN	O	O
yet	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
rapamycin	NN	O	O
on	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
p70	NN	O	B-protein
(	NN	O	I-protein
s6k	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
activation	NN	O	O
of	NN	O	O
E2Fs	NN	O	B-protein
have	NN	O	O
not	NN	O	O
been	NN	O	O
characterized	NN	O	O
.	NN	O	O

Our	NN	O	O
present	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
-and	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
-induced	NN	O	O
pathways	NN	O	O
for	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
include	NN	O	O
both	NN	O	O
rapamycin-resistant	NN	O	O
and	NN	O	O
rapamycin-sensitive	NN	O	O
components	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
a	NN	O	O
rapamycin-resistant	NN	O	B-protein
mutant	NN	O	I-protein
of	NN	O	O
p70	NN	O	B-protein
(	NN	O	I-protein
s6k	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
could	NN	O	O
restore	NN	O	O
rapamycin-suppressed	NN	O	O
E2F	NN	O	B-protein
responses	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
rapamycin-controlled	NN	O	O
processes	NN	O	O
involved	NN	O	O
in	NN	O	O
E2F	NN	O	B-protein
regulation	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
p70	NN	O	B-protein
(	NN	O	I-protein
s6k	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
rapamycin-resistant	NN	O	B-protein
p70	NN	O	I-protein
(	NN	O	I-protein
s6k	NN	O	I-protein
)	NN	O	I-protein
could	NN	O	O
not	NN	O	O
rescue	NN	O	O
rapamycin	NN	O	O
inhibition	NN	O	O
of	NN	O	O
T-cell	NN	O	O
cycle	NN	O	O
entry	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
additional	NN	O	O
,	NN	O	O
rapamycin-sensitive	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
T-cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
results	NN	O	O
thus	NN	O	O
show	NN	O	O
that	NN	O	O
p70	NN	O	B-protein
(	NN	O	I-protein
s6k	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
regulate	NN	O	O
E2F	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
and	NN	O	O
provide	NN	O	O
direct	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
for	NN	O	O
a	NN	O	O
link	NN	O	O
between	NN	O	O
IL-2	NN	O	B-protein
receptors	NN	O	O
,	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
p70	NN	O	B-protein
(	NN	O	I-protein
s6k	NN	O	I-protein
)	NN	O	I-protein
that	NN	O	O
regulates	NN	O	O
a	NN	O	O
crucial	NN	O	O
G1	NN	O	O
checkpoint	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Oxidative	NN	O	O
stress	NN	O	O
triggers	NN	O	O
STAT3	NN	O	B-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Oxidizing	NN	O	O
agents	NN	O	O
are	NN	O	O
powerful	NN	O	O
activators	NN	O	O
of	NN	O	O
factors	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
cytokine-encoding	NN	O	B-DNA
genes	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
tissue	NN	O	O
injury	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
show	NN	O	O
evidence	NN	O	O
that	NN	O	O
STAT3	NN	O	B-protein
is	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
whose	NN	O	O
activity	NN	O	O
is	NN	O	O
modulated	NN	O	O
by	NN	O	O
H2O2	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
in	NN	O	O
which	NN	O	O
endogenous	NN	O	O
catalase	NN	O	B-protein
had	NN	O	O
previously	NN	O	O
been	NN	O	O
inhibited	NN	O	O
.	NN	O	O

H2O2-induced	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
to	NN	O	O
form	NN	O	O
sequence-specific	NN	O	B-protein
DNA-bound	NN	O	I-protein
complexes	NN	O	I-protein
was	NN	O	O
evidenced	NN	O	O
by	NN	O	O
immunoblotting	NN	O	O
of	NN	O	O
nuclear	NN	O	O
fractions	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
and	NN	O	O
vanadate	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
strongly	NN	O	O
synergize	NN	O	O
with	NN	O	O
H2O2	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
anti-	NN	O	O
STAT3	NN	O	B-protein
antibodies	NN	O	I-protein
specifically	NN	O	O
precipitated	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
92	NN	O	B-protein
kDa	NN	O	I-protein
that	NN	O	O
becomes	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
upon	NN	O	O
lymphocyte	NN	O	O
treatment	NN	O	O
with	NN	O	O
H2O2	NN	O	O
.	NN	O	O

Phenylarsine	NN	O	O
oxide	NN	O	O
,	NN	O	O
a	NN	O	O
tyrosine	NN	O	O
phosphatase	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
and	NN	O	O
genistein	NN	O	O
,	NN	O	O
a	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
cooperated	NN	O	O
and	NN	O	O
cancelled	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
the	NN	O	O
H2O2-promoted	NN	O	O
STAT3	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

Evidence	NN	O	O
is	NN	O	O
also	NN	O	O
presented	NN	O	O
,	NN	O	O
using	NN	O	O
Fe2+/Cu2+	NN	O	O
ions	NN	O	O
,	NN	O	O
that.OH	NN	O	O
generated	NN	O	O
from	NN	O	O
H2O2	NN	O	O
through	NN	O	O
Fenton	NN	O	O
reactions	NN	O	O
could	NN	O	O
be	NN	O	O
a	NN	O	O
candidate	NN	O	O
oxygen	NN	O	O
reactive	NN	O	O
species	NN	O	O
to	NN	O	O
directly	NN	O	O
activate	NN	O	O
STAT3	NN	O	B-protein
.	NN	O	O

Present	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
H2O2	NN	O	O
and	NN	O	O
vanadate	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
intracellular	NN	O	B-protein
tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
enhanced	NN	O	O
STAT3	NN	O	B-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
hence	NN	O	O
its	NN	O	O
translocation	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
can	NN	O	O
be	NN	O	O
modulated	NN	O	O
by	NN	O	O
oxidizing	NN	O	O
agents	NN	O	O
and	NN	O	O
provide	NN	O	O
a	NN	O	O
framework	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
on	NN	O	O
the	NN	O	O
JAK	NN	O	B-protein
-STAT	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
and	NN	O	O
function	NN	O	O
.	NN	O	O

The	NN	O	O
development	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
are	NN	O	O
regulated	NN	O	O
tightly	NN	O	O
by	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
that	NN	O	O
include	NN	O	O
specific	NN	O	O
cell-surface	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
intracellular	NN	O	B-protein
signaling	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Since	NN	O	O
1988	NN	O	O
,	NN	O	O
several	NN	O	O
families	NN	O	O
of	NN	O	O
functionally	NN	O	O
important	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
have	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

These	NN	O	O
include	NN	O	O
the	NN	O	O
Ikaros	NN	O	B-protein
,	NN	O	O
LKLF	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GATA3	NN	O	B-protein
zinc-finger	NN	O	I-protein
proteins	NN	O	I-protein
;	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
,	NN	O	O
CREB/ATF	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel/NFAT	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
;	NN	O	O
the	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
;	NN	O	O
and	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
LEF1	NN	O	B-protein
,	NN	O	O
TCF1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Sox4	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
,	NN	O	O
we	NN	O	O
summarize	NN	O	O
our	NN	O	O
current	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
and	NN	O	O
function	NN	O	O
with	NN	O	O
particular	NN	O	O
emphasis	NN	O	O
on	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
recent	NN	O	O
gene	NN	O	O
targeting	NN	O	O
and	NN	O	O
transgenic	NN	O	O
experiments	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
increasing	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
pathways	NN	O	O
that	NN	O	O
regulate	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
and	NN	O	O
function	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
have	NN	O	O
suggested	NN	O	O
novel	NN	O	O
targets	NN	O	O
for	NN	O	O
genetic	NN	O	O
and	NN	O	O
pharmacological	NN	O	O
manipulation	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
immunity	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
T-lineage	NN	O	B-cell_type
cells	NN	O	I-cell_type
leads	NN	O	O
to	NN	O	O
a	NN	O	O
dramatic	NN	O	O
decrease	NN	O	O
in	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
and	NN	O	O
to	NN	O	O
increased	NN	O	O
cell	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogenic	NN	O	O
stimuli	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
to	NN	O	O
abnormal	NN	O	O
thymopoiesis	NN	O	O
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complexes	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
and	NN	O	O
activation	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
generated	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
in	NN	O	O
which	NN	O	O
RelA	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
complexes	NN	O	I-protein
were	NN	O	O
selectively	NN	O	O
inhibited	NN	O	O
in	NN	O	O
the	NN	O	O
T-lineage	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
specific	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
trans-dominant	NN	O	O
form	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

Transgene	NN	O	O
expression	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
thymic	NN	O	O
development	NN	O	O
,	NN	O	O
but	NN	O	O
led	NN	O	O
to	NN	O	O
lowered	NN	O	O
numbers	NN	O	O
of	NN	O	O
splenic	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
to	NN	O	O
a	NN	O	O
dramatic	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
ex	NN	O	O
vivo	NN	O	O
proliferative	NN	O	O
response	NN	O	O
of	NN	O	O
splenic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
perturbation	NN	O	O
of	NN	O	O
the	NN	O	O
proliferation	NN	O	O
response	NN	O	O
was	NN	O	O
not	NN	O	O
attributable	NN	O	O
to	NN	O	O
an	NN	O	O
abnormal	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
was	NN	O	O
strongly	NN	O	O
inhibited	NN	O	O
in	NN	O	O
the	NN	O	O
transgenic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
proliferative	NN	O	O
deficiency	NN	O	O
of	NN	O	O
the	NN	O	O
transgenic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
point	NN	O	O
out	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
growth	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
by	NN	O	O
either	NN	O	O
regulating	NN	O	B-protein
proteins	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
signaling	NN	O	O
or	NN	O	O
by	NN	O	O
functionally	NN	O	O
interfering	NN	O	O
with	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Paradoxical	NN	O	O
priming	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
on	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
is	NN	O	O
a	NN	O	O
well-known	NN	O	O
immunosuppressive	NN	O	B-protein
and/or	NN	O	I-protein
anti-inflammatory	NN	O	I-protein
cytokine	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
in	NN	O	O
vitro	NN	O	O
experimental	NN	O	O
studies	NN	O	O
in	NN	O	O
which	NN	O	O
IL-10	NN	O	B-cell_line
primed	NN	O	I-cell_line
leukocytes	NN	O	I-cell_line
and	NN	O	O
led	NN	O	O
to	NN	O	O
an	NN	O	O
enhanced	NN	O	O
production	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
upon	NN	O	O
further	NN	O	O
stimulation	NN	O	O
by	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

Monocytes	NN	O	B-cell_type
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
prepared	NN	O	O
from	NN	O	O
whole	NN	O	O
blood	NN	O	O
maintained	NN	O	O
for	NN	O	O
20	NN	O	O
h	NN	O	O
at	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
recombinant	NN	O	B-protein
human	NN	O	I-protein
IL-10	NN	O	I-protein
had	NN	O	O
an	NN	O	O
enhanced	NN	O	O
capacity	NN	O	O
to	NN	O	O
produce	NN	O	O
TNF	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
LPS-induced	NN	O	O
IL-6	NN	O	O
and	NN	O	O
spontaneous	NN	O	O
IL-1ra	NN	O	O
production	NN	O	O
were	NN	O	O
also	NN	O	O
enhanced	NN	O	O
.	NN	O	O

When	NN	O	O
isolated	NN	O	O
PBMC	NN	O	B-cell_type
were	NN	O	O
first	NN	O	O
cultured	NN	O	O
for	NN	O	O
20	NN	O	O
h	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
on	NN	O	O
Teflon	NN	O	O
to	NN	O	O
prevent	NN	O	O
adherence	NN	O	O
,	NN	O	O
washed	NN	O	O
to	NN	O	O
remove	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
then	NN	O	O
further	NN	O	O
cultured	NN	O	O
in	NN	O	O
plastic	NN	O	O
dishes	NN	O	O
for	NN	O	O
an	NN	O	O
additional	NN	O	O
20	NN	O	O
h	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
LPS	NN	O	O
or	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
an	NN	O	O
enhanced	NN	O	O
release	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
not	NN	O	O
the	NN	O	O
case	NN	O	O
when	NN	O	O
PBMC	NN	O	B-cell_type
were	NN	O	O
pre-cultured	NN	O	O
in	NN	O	O
plastic	NN	O	O
multidishes	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

TNF	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
LPS	NN	O	O
was	NN	O	O
decreased	NN	O	O
when	NN	O	O
the	NN	O	O
pre-treatment	NN	O	O
of	NN	O	O
PBMC	NN	O	B-cell_type
with	NN	O	O
IL-10	NN	O	B-protein
was	NN	O	O
performed	NN	O	O
on	NN	O	O
plastic	NN	O	O
,	NN	O	O
whereas	NN	O	O
this	NN	O	O
was	NN	O	O
not	NN	O	O
the	NN	O	O
case	NN	O	O
when	NN	O	O
cells	NN	O	O
were	NN	O	O
pre-cultured	NN	O	O
with	NN	O	O
IL-10	NN	O	B-protein
on	NN	O	O
Teflon	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
NFkappaB	NN	O	O
translocation	NN	O	O
following	NN	O	O
LPS	NN	O	O
activation	NN	O	O
was	NN	O	O
higher	NN	O	O
after	NN	O	O
IL-10	NN	O	B-protein
pre-treatment	NN	O	O
on	NN	O	O
Teflon	NN	O	O
than	NN	O	O
on	NN	O	O
plastic	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
an	NN	O	O
enhanced	NN	O	O
frequency	NN	O	O
of	NN	O	O
CD16	NN	O	B-protein
and	NN	O	O
CD68	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
cells	NN	O	O
among	NN	O	O
the	NN	O	O
CD14	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
cells	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
independently	NN	O	O
of	NN	O	O
the	NN	O	O
pre-culture	NN	O	O
conditions	NN	O	O
of	NN	O	O
the	NN	O	O
PBMC	NN	O	B-cell_type
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
IL-10	NN	O	B-protein
-induced	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
prevention	NN	O	O
of	NN	O	O
monocyte	NN	O	O
adherence	NN	O	O
by	NN	O	O
red	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
whole	NN	O	O
blood	NN	O	O
assays	NN	O	O
or	NN	O	O
by	NN	O	O
cultures	NN	O	O
of	NN	O	O
PBMC	NN	O	B-cell_type
on	NN	O	O
Teflon	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
adherence	NN	O	O
parameter	NN	O	O
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
IL-10	NN	O	B-protein
-induced	NN	O	O
modulation	NN	O	O
of	NN	O	O
some	NN	O	O
monocyte	NN	O	B-protein
surface	NN	O	I-protein
markers	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Cell	NN	O	O
growth-regulated	NN	O	O
expression	NN	O	O
of	NN	O	O
mammalian	NN	O	B-DNA
MCM5	NN	O	I-DNA
and	NN	O	I-DNA
MCM6	NN	O	I-DNA
genes	NN	O	I-DNA
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
E2F	NN	O	B-protein
.	NN	O	O

Initiation	NN	O	O
of	NN	O	O
DNA	NN	O	O
replication	NN	O	O
requires	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
MCM	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
,	NN	O	O
which	NN	O	O
participate	NN	O	O
in	NN	O	O
ensuring	NN	O	O
that	NN	O	O
DNA	NN	O	O
replication	NN	O	O
occurs	NN	O	O
only	NN	O	O
once	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
all	NN	O	O
mammalian	NN	O	B-DNA
genes	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
MCM	NN	O	B-DNA
family	NN	O	I-DNA
is	NN	O	O
induced	NN	O	O
by	NN	O	O
growth	NN	O	O
stimulation	NN	O	O
,	NN	O	O
unlike	NN	O	O
yeast	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
peak	NN	O	O
at	NN	O	O
G1/S	NN	O	O
boundary	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
of	NN	O	O
isolated	NN	O	B-DNA
human	NN	O	I-DNA
MCM	NN	O	I-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

Human	NN	O	B-DNA
MCM5	NN	O	I-DNA
and	NN	O	I-DNA
MCM6	NN	O	I-DNA
promoters	NN	O	I-DNA
with	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
E2F	NN	O	B-DNA
sites	NN	O	I-DNA
failed	NN	O	O
in	NN	O	O
promoter	NN	O	O
regulation	NN	O	O
following	NN	O	O
serum	NN	O	O
stimulation	NN	O	O
and	NN	O	O
exogenous	NN	O	O
E2F	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	O
E2F-like	NN	O	B-DNA
sequence	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-DNA
MCM6	NN	O	I-DNA
promoter	NN	O	I-DNA
which	NN	O	O
cooperates	NN	O	O
with	NN	O	O
the	NN	O	O
authentic	NN	O	O
E2F	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
E2F	NN	O	B-protein
-dependent	NN	O	O
regulation	NN	O	O
.	NN	O	O

Forced	NN	O	O
expression	NN	O	O
of	NN	O	O
E2F1	NN	O	B-protein
could	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
all	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
MCM	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
rat	NN	O	B-cell_line
embryonal	NN	O	I-cell_line
fibroblast	NN	O	I-cell_line
REF52	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
growth-regulated	NN	O	O
expression	NN	O	O
of	NN	O	O
mammalian	NN	O	B-DNA
MCM5	NN	O	I-DNA
and	NN	O	I-DNA
MCM6	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
presumably	NN	O	O
other	NN	O	O
MCM	NN	O	O
members	NN	O	O
,	NN	O	O
is	NN	O	O
primarily	NN	O	O
regulated	NN	O	O
by	NN	O	O
E2F	NN	O	B-protein
through	NN	O	O
binding	NN	O	O
to	NN	O	O
multiple	NN	O	B-DNA
E2F	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
essential	NN	O	O
role	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
CD34	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cell	NN	O	I-cell_type
survival	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
B-cell	NN	O	O
maturation	NN	O	O
,	NN	O	O
and	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
transcription	NN	O	O
and	NN	O	O
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
alternatively	NN	O	O
mediating	NN	O	O
and	NN	O	O
protecting	NN	O	O
against	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
CD34	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
not	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

We	NN	O	O
provide	NN	O	O
evidence	NN	O	O
here	NN	O	O
that	NN	O	O
virtually	NN	O	O
all	NN	O	O
human	NN	O	B-cell_type
CD34	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
NF-kappaB	NN	O	B-protein
that	NN	O	O
can	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
exposure	NN	O	O
to	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
and	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
eg	NN	O	O
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
interleukin-3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
may	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
human	NN	O	O
CD34	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cell	NN	O	I-cell_type
clonogenic	NN	O	O
function	NN	O	O
and	NN	O	O
survival	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
offer	NN	O	O
insight	NN	O	O
into	NN	O	O
a	NN	O	O
new	NN	O	O
role	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
maintaining	NN	O	O
survival	NN	O	O
and	NN	O	O
function	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
stem	NN	O	I-cell_type
and	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
proposed	NN	O	O
strategies	NN	O	O
involving	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
as	NN	O	O
an	NN	O	O
adjunct	NN	O	O
to	NN	O	O
cancer	NN	O	O
chemotherapy	NN	O	O
should	NN	O	O
be	NN	O	O
approached	NN	O	O
with	NN	O	O
caution	NN	O	O
.	NN	O	O

-DOCSTART-	O

High	NN	O	O
molecular	NN	O	O
weight	NN	O	O
dextran	NN	O	O
sulfate	NN	O	O
increases	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappaB-regulated	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
is	NN	O	O
known	NN	O	O
that	NN	O	O
sulfated	NN	O	O
polysaccharides	NN	O	O
can	NN	O	O
mimic	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
common	NN	O	B-protein
T-cell	NN	O	I-protein
mitogens	NN	O	I-protein
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
mitogenic	NN	O	O
effect	NN	O	O
of	NN	O	O
high	NN	O	O
molecular	NN	O	O
weight	NN	O	O
dextran	NN	O	O
sulfate	NN	O	O
(	NN	O	O
HMDS	NN	O	O
)	NN	O	O
,	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
were	NN	O	O
transfected	NN	O	O
with	NN	O	O
recombinant	NN	O	B-DNA
plasmid	NN	O	I-DNA
containing	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyl	NN	O	I-DNA
transferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	I-DNA
LTR	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
HMDS	NN	O	O
,	NN	O	O
similar	NN	O	O
to	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
increases	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
suggesting	NN	O	O
increased	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
B7.1	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

It	NN	O	O
was	NN	O	O
postulated	NN	O	O
that	NN	O	O
this	NN	O	O
NF-kappaB-regulated	NN	O	B-DNA
promoter	NN	O	I-DNA
might	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
accessory	NN	O	O
cells	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
replication	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
Sp1	NN	O	B-protein
and	NN	O	O
AP-2	NN	O	B-protein
mediate	NN	O	O
induction	NN	O	O
of	NN	O	O
acid	NN	O	B-protein
sphingomyelinase	NN	O	I-protein
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Cells	NN	O	O
from	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	B-cell_line
differentiate	NN	O	O
towards	NN	O	O
a	NN	O	O
macrophage-like	NN	O	O
phenotype	NN	O	O
when	NN	O	O
stimulated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
-13-	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxy-vitamin	NN	O	O
D3	NN	O	O
,	NN	O	O
and	NN	O	O
various	NN	O	O
other	NN	O	O
agents	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
lysosomal	NN	O	B-protein
enzyme	NN	O	I-protein
acid	NN	O	B-protein
sphingomyelinase	NN	O	I-protein
(	NN	O	O
ASM	NN	O	O
;	NN	O	O
E.C.3.1.4.12	NN	O	O
)	NN	O	O
is	NN	O	O
induced	NN	O	O
during	NN	O	O
this	NN	O	O
process	NN	O	O
and	NN	O	O
is	NN	O	O
strongly	NN	O	O
elevated	NN	O	O
in	NN	O	O
differentiated	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
differentiated	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
phagocytes	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
Northern	NN	O	O
blotting	NN	O	O
,	NN	O	O
RNase	NN	O	B-protein
protection	NN	O	O
assay	NN	O	O
,	NN	O	O
and	NN	O	O
nuclear	NN	O	O
run-on	NN	O	O
analyses	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
ASM	NN	O	O
expression	NN	O	O
is	NN	O	O
regulated	NN	O	O
mainly	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
and	NN	O	O
that	NN	O	O
new	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
enhanced	NN	O	O
ASM	NN	O	O
activity	NN	O	O
.	NN	O	O

This	NN	O	O
cell-type	NN	O	O
specific	NN	O	O
induction	NN	O	O
by	NN	O	O
PMA	NN	O	O
treatment	NN	O	O
was	NN	O	O
further	NN	O	O
investigated	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
.	NN	O	O

A	NN	O	O
series	NN	O	O
of	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
deletion	NN	O	I-DNA
derivatives	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
were	NN	O	O
used	NN	O	O
in	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
to	NN	O	O
identify	NN	O	O
promoter	NN	O	B-DNA
elements	NN	O	I-DNA
required	NN	O	O
for	NN	O	O
basal	NN	O	O
and	NN	O	O
inducible	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

A	NN	O	O
PMA	NN	O	B-DNA
responsive	NN	O	I-DNA
element	NN	O	I-DNA
was	NN	O	O
localized	NN	O	O
to	NN	O	O
a	NN	O	O
region	NN	O	O
between	NN	O	O
-319	NN	O	B-DNA
and	NN	O	I-DNA
-219	NN	O	I-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
initiation	NN	O	O
codon	NN	O	O
and	NN	O	O
co-transfections	NN	O	O
with	NN	O	O
transcription	NN	O	B-DNA
factor	NN	O	I-DNA
expression	NN	O	I-DNA
plasmids	NN	O	I-DNA
for	NN	O	O
AP-2	NN	O	B-protein
and	NN	O	O
Sp1	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
augmented	NN	O	O
ASM	NN	O	O
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
abolished	NN	O	O
when	NN	O	O
the	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
these	NN	O	O
two	NN	O	O
factors	NN	O	O
were	NN	O	O
deleted	NN	O	O
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
and	NN	O	O
supershift	NN	O	O
assays	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
this	NN	O	O
region	NN	O	O
is	NN	O	O
specifically	NN	O	O
bound	NN	O	O
by	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
AP-2	NN	O	B-protein
.	NN	O	O

These	NN	O	O
factors	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
from	NN	O	O
both	NN	O	O
induced	NN	O	O
and	NN	O	O
uninduced	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
intensity	NN	O	O
of	NN	O	O
the	NN	O	O
complex	NN	O	O
formed	NN	O	O
appeared	NN	O	O
to	NN	O	O
increase	NN	O	O
when	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
PMA-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
used	NN	O	O
.	NN	O	O

From	NN	O	O
these	NN	O	O
studies	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
a	NN	O	O
concerted	NN	O	O
action	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP-2	NN	O	B-protein
and	NN	O	O
Sp1	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
up	NN	O	O
-regulation	NN	O	O
of	NN	O	O
ASM	NN	O	O
expression	NN	O	O
during	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

LPS-Induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
are	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
dependent	NN	O	O
and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
independent	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
mediator	NN	O	O
of	NN	O	O
septic	NN	O	O
shock	NN	O	O
.	NN	O	O

Endotoxin	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
signal	NN	O	O
transduction	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
leads	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
implicated	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
and	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
(	NN	O	O
PTK	NN	O	B-protein
)	NN	O	O
in	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
inhibition	NN	O	O
of	NN	O	O
either	NN	O	O
PKC	NN	O	B-protein
or	NN	O	O
PTK	NN	O	B-protein
would	NN	O	O
decrease	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

MATERIALS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
Human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
PMA	NN	O	O
(	NN	O	O
50	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
alone	NN	O	O
or	NN	O	O
LPS	NN	O	O
(	NN	O	O
100	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
with	NN	O	O
and	NN	O	O
without	NN	O	O
a	NN	O	O
nonspecific	NN	O	O
serine/threonine	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
inhibitor	NN	O	O
staurosporine	NN	O	O
(	NN	O	O
Stauro	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
specific	NN	O	O
pan-	NN	O	O
PKC	NN	O	B-protein
inhibitor	NN	O	O
bisindolylmaleimide	NN	O	O
(	NN	O	O
Bis	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
PTK	NN	O	B-protein
genistein	NN	O	O
(	NN	O	O
Gen	NN	O	O
)	NN	O	O
.	NN	O	O

TNF-alpha	NN	O	B-protein
release	NN	O	O
in	NN	O	O
culture	NN	O	O
supernatants	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
an	NN	O	O
ELISA	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
was	NN	O	O
evaluated	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
LPS	NN	O	O
increased	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Nonspecific	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
inhibition	NN	O	O
inhibited	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
,	NN	O	O
while	NN	O	O
specific	NN	O	O
PKC	NN	O	B-protein
inhibition	NN	O	O
with	NN	O	O
Bis	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
or	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
.	NN	O	O

PTK	NN	O	B-protein
inhibition	NN	O	O
with	NN	O	O
Gen	NN	O	O
attenuated	NN	O	O
both	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Direct	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
with	NN	O	O
PMA	NN	O	O
induced	NN	O	O
both	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
are	NN	O	O
independent	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
PTK	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
thus	NN	O	O
could	NN	O	O
be	NN	O	O
a	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
target	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
states	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Amelioration	NN	O	O
of	NN	O	O
rat	NN	O	O
cerulein	NN	O	O
pancreatitis	NN	O	O
by	NN	O	O
guamerin-derived	NN	O	O
peptide	NN	O	O
,	NN	O	O
a	NN	O	O
novel	NN	O	O
elastase	NN	O	B-protein
inhibitor	NN	O	O
.	NN	O	O

Increased	NN	O	O
activity	NN	O	O
of	NN	O	O
various	NN	O	O
proteases	NN	O	B-protein
is	NN	O	O
observed	NN	O	O
in	NN	O	O
both	NN	O	O
human	NN	O	O
and	NN	O	O
experimental	NN	O	O
pancreatitis	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
information	NN	O	O
on	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
specific	NN	O	O
protease	NN	O	O
inhibitors	NN	O	O
on	NN	O	O
the	NN	O	O
disease	NN	O	O
is	NN	O	O
limited	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
novel	NN	O	O
elastase	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
guamerin-derived	NN	O	O
synthetic	NN	O	O
peptide	NN	O	O
(	NN	O	O
GDSP	NN	O	O
)	NN	O	O
,	NN	O	O
improves	NN	O	O
the	NN	O	O
parameters	NN	O	O
of	NN	O	O
cerulein-induced	NN	O	O
acute	NN	O	O
pancreatitis	NN	O	O
in	NN	O	O
the	NN	O	O
rat	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
GDSP	NN	O	O
on	NN	O	O
pancreatic	NN	O	O
weight	NN	O	O
,	NN	O	O
serum	NN	O	B-protein
amylase	NN	O	I-protein
and	NN	O	I-protein
lipase	NN	O	I-protein
,	NN	O	O
morphologic	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
pancreas	NN	O	O
,	NN	O	O
neutrophil	NN	O	O
infiltration	NN	O	O
,	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
KB	NN	O	I-protein
(	NN	O	O
NF-KB	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
were	NN	O	O
measured	NN	O	O
in	NN	O	O
rats	NN	O	O
infused	NN	O	O
with	NN	O	O
supramaximal	NN	O	O
dose	NN	O	O
of	NN	O	O
cerulein	NN	O	O
(	NN	O	O
5	NN	O	O
(	NN	O	O
g/kg/h	NN	O	O
)	NN	O	O
for	NN	O	O
6	NN	O	O
h	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
GDSP	NN	O	O
were	NN	O	O
also	NN	O	O
measured	NN	O	O
on	NN	O	O
superoxide	NN	O	O
formation	NN	O	O
by	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
GDSP	NN	O	O
were	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
another	NN	O	O
elastase	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
elastatinal	NN	O	O
.	NN	O	O

GDSP	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
edema	NN	O	O
formation	NN	O	O
,	NN	O	O
neutrophil	NN	O	O
infiltration	NN	O	O
,	NN	O	O
acinar	NN	O	O
cell	NN	O	O
damage	NN	O	O
,	NN	O	O
and	NN	O	O
plasma	NN	O	B-protein
lipase	NN	O	I-protein
and	NN	O	O
amylase	NN	O	B-protein
increases	NN	O	O
caused	NN	O	O
by	NN	O	O
cerulein	NN	O	O
.	NN	O	O

GDSP	NN	O	O
also	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
superoxide	NN	O	O
formation	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
N-formyl-methionine-leucine-phenyl-alanine	NN	O	O
(	NN	O	O
fMLP	NN	O	O
)	NN	O	O
or	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
.	NN	O	O

Elastatinal	NN	O	O
had	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
effects	NN	O	O
as	NN	O	O
GDSP	NN	O	O
but	NN	O	O
was	NN	O	O
less	NN	O	O
potent	NN	O	O
and	NN	O	O
effective	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
beneficial	NN	O	O
effect	NN	O	O
of	NN	O	O
GDSP	NN	O	O
,	NN	O	O
a	NN	O	O
novel	NN	O	O
specific	NN	O	O
elastase	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
rat	NN	O	O
cerulein	NN	O	O
pancreatitis	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
lipopolysaccharide-induced	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
regulating	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
:	NN	O	O
molecular	NN	O	O
cloning	NN	O	O
,	NN	O	O
sequencing	NN	O	O
,	NN	O	O
characterization	NN	O	O
,	NN	O	O
and	NN	O	O
chromosomal	NN	O	O
assignment	NN	O	O
.	NN	O	O

Lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
stimulator	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
causing	NN	O	O
secretion	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
other	NN	O	O
inflammatory	NN	O	O
mediators	NN	O	O
.	NN	O	O

Given	NN	O	O
the	NN	O	O
deleterious	NN	O	O
effects	NN	O	O
to	NN	O	O
the	NN	O	O
host	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
postulated	NN	O	O
that	NN	O	O
TNF-alpha	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
must	NN	O	O
be	NN	O	O
tightly	NN	O	O
regulated	NN	O	O
.	NN	O	O

The	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
that	NN	O	O
control	NN	O	O
TNF-alpha	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
humans	NN	O	O
remains	NN	O	O
obscure	NN	O	O
,	NN	O	O
although	NN	O	O
NF-kappaB	NN	O	B-protein
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
studies	NN	O	O
pertaining	NN	O	O
to	NN	O	O
macrophage	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
located	NN	O	O
from	NN	O	O
-550	NN	O	O
to	NN	O	O
-487	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
TNF-alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
that	NN	O	O
contains	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
,	NN	O	O
but	NN	O	O
lacks	NN	O	O
any	NN	O	O
known	NN	O	O
NF-kappaB	NN	O	B-protein
-binding	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
this	NN	O	O
DNA	NN	O	O
fragment	NN	O	O
to	NN	O	O
isolate	NN	O	O
and	NN	O	O
purify	NN	O	O
a	NN	O	O
60-kDa	NN	O	B-protein
protein	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
this	NN	O	O
fragment	NN	O	O
and	NN	O	O
obtained	NN	O	O
its	NN	O	O
amino-terminal	NN	O	B-protein
sequence	NN	O	I-protein
,	NN	O	O
which	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
design	NN	O	O
degenerate	NN	O	B-DNA
probes	NN	O	I-DNA
to	NN	O	O
screen	NN	O	O
a	NN	O	O
cDNA	NN	O	O
library	NN	O	O
from	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
novel	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
(	NN	O	O
1.8	NN	O	O
kb	NN	O	O
)	NN	O	O
was	NN	O	O
isolated	NN	O	O
and	NN	O	O
fully	NN	O	O
sequenced	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
this	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
revealed	NN	O	O
that	NN	O	O
its	NN	O	O
induction	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
LPS	NN	O	O
activation	NN	O	O
of	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
;	NN	O	O
hence	NN	O	O
,	NN	O	O
the	NN	O	O
name	NN	O	O
LPS-induced	NN	O	B-protein
TNF-alpha	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
LITAF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
LITAF	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
reduction	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
transcripts	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
LITAF	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
observed	NN	O	O
predominantly	NN	O	O
in	NN	O	O
the	NN	O	O
placenta	NN	O	O
,	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
spleen	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
chromosomal	NN	O	O
localization	NN	O	O
using	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
revealed	NN	O	O
that	NN	O	O
LITAF	NN	O	B-protein
mapped	NN	O	O
to	NN	O	O
chromosome	NN	O	B-DNA
16p12-16p13.3	NN	O	I-DNA
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
LITAF	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
TNF-alpha	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
proposes	NN	O	O
a	NN	O	O
new	NN	O	O
mechanism	NN	O	O
to	NN	O	O
control	NN	O	O
TNF-alpha	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
human	NN	O	B-DNA
IFNGR1	NN	O	I-DNA
small	NN	O	I-DNA
deletion	NN	O	I-DNA
hotspot	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
dominant	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
mycobacterial	NN	O	O
infection	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

The	NN	O	O
immunogenetic	NN	O	O
basis	NN	O	O
of	NN	O	O
severe	NN	O	O
infections	NN	O	O
caused	NN	O	O
by	NN	O	O
bacille	NN	O	O
Calmette-Guerin	NN	O	O
vaccine	NN	O	O
and	NN	O	O
environmental	NN	O	O
mycobacteria	NN	O	O
in	NN	O	O
humans	NN	O	O
remains	NN	O	O
largely	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
18	NN	O	O
patients	NN	O	O
from	NN	O	O
several	NN	O	O
generations	NN	O	O
of	NN	O	O
12	NN	O	O
unrelated	NN	O	O
families	NN	O	O
who	NN	O	O
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
1	NN	O	O
to	NN	O	O
5	NN	O	O
overlapping	NN	O	B-DNA
IFNGR1	NN	O	I-DNA
frameshift	NN	O	I-DNA
small	NN	O	I-DNA
deletions	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
wild-type	NN	O	B-DNA
IFNGR1	NN	O	I-DNA
allele	NN	O	I-DNA
.	NN	O	O

There	NN	O	O
were	NN	O	O
12	NN	O	O
independent	NN	O	O
mutation	NN	O	O
events	NN	O	O
at	NN	O	O
a	NN	O	O
single	NN	O	B-DNA
mutation	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
defining	NN	O	O
a	NN	O	O
small	NN	O	B-DNA
deletion	NN	O	I-DNA
hotspot	NN	O	I-DNA
.	NN	O	O

Neighbouring	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
favours	NN	O	O
a	NN	O	O
small	NN	O	O
deletion	NN	O	O
model	NN	O	O
of	NN	O	O
slipped	NN	O	O
mispairing	NN	O	O
events	NN	O	O
during	NN	O	O
replication	NN	O	O
.	NN	O	O

The	NN	O	O
mutant	NN	O	O
alleles	NN	O	O
encode	NN	O	O
cell-surface	NN	O	B-protein
IFNgamma	NN	O	I-protein
receptors	NN	O	I-protein
that	NN	O	O
lack	NN	O	O
the	NN	O	O
intra-cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
which	NN	O	O
,	NN	O	O
through	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
impaired	NN	O	O
recycling	NN	O	O
,	NN	O	O
abrogated	NN	O	O
signalling	NN	O	O
and	NN	O	O
normal	NN	O	O
binding	NN	O	O
to	NN	O	O
IFNgamma	NN	O	B-protein
exert	NN	O	O
a	NN	O	O
dominant-negative	NN	O	O
effect	NN	O	O
.	NN	O	O

We	NN	O	O
thus	NN	O	O
report	NN	O	O
a	NN	O	O
hotspot	NN	O	O
for	NN	O	O
human	NN	O	B-DNA
IFNGR1	NN	O	I-DNA
small	NN	O	I-DNA
deletions	NN	O	I-DNA
that	NN	O	O
confer	NN	O	O
dominant	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
infections	NN	O	O
caused	NN	O	O
by	NN	O	O
poorly	NN	O	O
virulent	NN	O	O
mycobacteria	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
human	NN	O	B-protein
pterin	NN	O	I-protein
carbinolamine	NN	O	I-protein
dehydratase/dimerization	NN	O	I-protein
cofactor	NN	O	I-protein
of	NN	O	O
HNF1	NN	O	B-protein
.	NN	O	O

Pterin	NN	O	B-protein
carbinolamine	NN	O	I-protein
dehydratase/dimerization	NN	O	I-protein
cofactor	NN	O	I-protein
of	NN	O	O
HNF1	NN	O	B-protein
(	NN	O	O
PCD/DCoH	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
dual-function	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
it	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
dehydratase	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regeneration	NN	O	O
of	NN	O	O
tetrahydrobiopterin	NN	O	B-protein
,	NN	O	O
the	NN	O	O
cofactor	NN	O	O
for	NN	O	O
aromatic	NN	O	B-protein
amino	NN	O	I-protein
acid	NN	O	I-protein
hydroxylases	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
it	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
dimerization	NN	O	B-protein
cofactor	NN	O	I-protein
of	NN	O	O
HNF1	NN	O	B-protein
and	NN	O	O
increases	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
HNF1	NN	O	B-protein
.	NN	O	O

To	NN	O	O
deepen	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
,	NN	O	O
we	NN	O	O
characterized	NN	O	O
its	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	O
tissues	NN	O	O
and	NN	O	O
cells	NN	O	O
.	NN	O	O

Human	NN	O	O
PCD/DCoH	NN	O	B-protein
was	NN	O	O
present	NN	O	O
predominantly	NN	O	O
in	NN	O	O
liver	NN	O	O
and	NN	O	O
kidney	NN	O	O
,	NN	O	O
with	NN	O	O
significant	NN	O	O
amounts	NN	O	O
in	NN	O	O
testis	NN	O	O
and	NN	O	O
ovary	NN	O	O
,	NN	O	O
trace	NN	O	O
amounts	NN	O	O
in	NN	O	O
lung	NN	O	O
,	NN	O	O
and	NN	O	O
undetectable	NN	O	O
levels	NN	O	O
in	NN	O	O
whole	NN	O	O
brain	NN	O	O
,	NN	O	O
heart	NN	O	O
,	NN	O	O
and	NN	O	O
spleen	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
that	NN	O	O
were	NN	O	O
examined	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
also	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
lack	NN	O	O
HNF1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
fibroblasts	NN	O	B-cell_type
that	NN	O	O
have	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
tetrahydrobiopterin	NN	O	B-protein
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-protein
PCD/DCoH	NN	O	I-protein
in	NN	O	O
the	NN	O	O
liver	NN	O	O
and	NN	O	O
nonhepatic	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
compared	NN	O	O
at	NN	O	O
both	NN	O	O
the	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
levels	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
in	NN	O	O
liver	NN	O	O
was	NN	O	O
only	NN	O	O
fourfold	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
keratinocytes	NN	O	B-cell_type
and	NN	O	O
fibroblasts	NN	O	B-cell_type
,	NN	O	O
the	NN	O	O
hepatic	NN	O	B-protein
PCD/DCoH	NN	O	I-protein
protein	NN	O	I-protein
level	NN	O	O
was	NN	O	O
20-fold	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
epidermal	NN	O	I-cell_type
keratinocytes	NN	O	I-cell_type
and	NN	O	O
dermal	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

Cloning	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
and	NN	O	I-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
untranslated	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
UTR	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
human	NN	O	B-protein
keratinocyte	NN	O	I-protein
PCD/DCoH	NN	O	I-protein
revealed	NN	O	O
that	NN	O	O
it	NN	O	O
has	NN	O	O
53	NN	O	O
bp	NN	O	O
more	NN	O	O
of	NN	O	O
GC-rich	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
untranslated	NN	O	I-DNA
sequence	NN	O	I-DNA
than	NN	O	O
the	NN	O	O
published	NN	O	O
liver	NN	O	B-protein
PCD/DCoH	NN	O	I-protein
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
and	NN	O	O
translation	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
longer	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
UTR	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
about	NN	O	O
a	NN	O	O
35	NN	O	O
%	NN	O	O
decrease	NN	O	O
in	NN	O	O
translation	NN	O	O
efficiency	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
human	NN	O	B-protein
PCD/DCoH	NN	O	I-protein
is	NN	O	O
not	NN	O	O
only	NN	O	O
present	NN	O	O
in	NN	O	O
cells	NN	O	O
where	NN	O	O
tetrahydrobiopterin	NN	O	B-protein
is	NN	O	O
synthesized	NN	O	O
or	NN	O	O
HNF1	NN	O	B-protein
is	NN	O	O
present	NN	O	O
but	NN	O	O
is	NN	O	O
a	NN	O	O
widely	NN	O	O
distributed	NN	O	O
protein	NN	O	O
.	NN	O	O

Its	NN	O	O
differential	NN	O	O
expression	NN	O	O
in	NN	O	O
different	NN	O	O
tissues	NN	O	O
and	NN	O	O
cells	NN	O	O
is	NN	O	O
regulated	NN	O	O
not	NN	O	O
only	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
but	NN	O	O
also	NN	O	O
at	NN	O	O
the	NN	O	O
translational	NN	O	O
level	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappaB	NN	O	B-protein
regulates	NN	O	O
Fas/APO-1/CD95-	NN	O	O
and	NN	O	O
TCR-	NN	O	O
mediated	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
maintenance	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
homeostasis	NN	O	O
by	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
regulatory	NN	O	O
mechanism	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
protecting	NN	O	O
cells	NN	O	O
against	NN	O	O
death	NN	O	O
mediated	NN	O	O
by	NN	O	O
TNF	NN	O	B-protein
We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
also	NN	O	O
has	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
Fas	NN	O	B-protein
/	NN	O	O
APO-1	NN	O	B-protein
/	NN	O	O
CD95	NN	O	B-protein
-mediated	NN	O	O
death	NN	O	O
,	NN	O	O
a	NN	O	O
major	NN	O	O
pathway	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
death	NN	O	O
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
subunits	NN	O	I-protein
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
confers	NN	O	O
resistance	NN	O	O
against	NN	O	O
Fas-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Reciprocally	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
a	NN	O	O
soluble	NN	O	O
peptide	NN	O	O
inhibitor	NN	O	O
or	NN	O	O
a	NN	O	O
dominant	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
inhibitor	NN	O	O
,	NN	O	O
IkappaB	NN	O	B-protein
,	NN	O	O
makes	NN	O	O
the	NN	O	O
cells	NN	O	O
more	NN	O	O
susceptible	NN	O	O
to	NN	O	O
Fas-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
a	NN	O	O
soluble	NN	O	O
peptide	NN	O	O
inhibitor	NN	O	O
rendered	NN	O	O
a	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridoma	NN	O	I-cell_line
more	NN	O	O
susceptible	NN	O	O
to	NN	O	O
TCR-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Correspondingly	NN	O	O
,	NN	O	O
transfection	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
provided	NN	O	O
considerable	NN	O	O
protection	NN	O	O
from	NN	O	O
TCR-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
were	NN	O	O
corroborated	NN	O	O
by	NN	O	O
studies	NN	O	O
on	NN	O	O
Fas	NN	O	B-protein
-mediated	NN	O	O
death	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Concanavalin	NN	O	O
A-activated	NN	O	O
cycling	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
blasts	NN	O	I-cell_line
from	NN	O	O
mice	NN	O	O
that	NN	O	O
are	NN	O	O
transgenic	NN	O	O
for	NN	O	O
the	NN	O	O
dominant	NN	O	B-protein
IkappaB	NN	O	I-protein
molecule	NN	O	I-protein
have	NN	O	O
increased	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
Fas-mediated	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
blocking	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
itself	NN	O	O
a	NN	O	O
positive	NN	O	O
regulator	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
with	NN	O	O
neutralizing	NN	O	B-protein
antibodies	NN	O	I-protein
renders	NN	O	O
the	NN	O	O
cells	NN	O	O
more	NN	O	O
susceptible	NN	O	O
to	NN	O	O
anti-Fas-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
provide	NN	O	O
compelling	NN	O	O
evidence	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
protects	NN	O	O
against	NN	O	O
Fas	NN	O	B-protein
-mediated	NN	O	O
death	NN	O	O
and	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
regulator	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
homeostasis	NN	O	O
and	NN	O	O
tolerance	NN	O	O
.	NN	O	O

-DOCSTART-	O

T	NN	O	O
helper	NN	O	O
differentiation	NN	O	O
proceeds	NN	O	O
through	NN	O	O
Stat1-dependent	NN	O	O
,	NN	O	O
Stat4-dependent	NN	O	O
and	NN	O	O
Stat4-independent	NN	O	O
phases	NN	O	O
.	NN	O	O

Much	NN	O	O
of	NN	O	O
our	NN	O	O
focus	NN	O	O
in	NN	O	O
understanding	NN	O	O
Th1/Th2	NN	O	O
development	NN	O	O
has	NN	O	O
been	NN	O	O
on	NN	O	O
the	NN	O	O
signals	NN	O	O
delivered	NN	O	O
by	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
as	NN	O	O
final	NN	O	O
determinants	NN	O	O
of	NN	O	O
terminal	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Because	NN	O	O
extinction	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
early	NN	O	O
Th2	NN	O	O
development	NN	O	O
could	NN	O	O
potentially	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
imprinting	NN	O	O
a	NN	O	O
more	NN	O	O
permanent	NN	O	O
Th2	NN	O	O
phenotype	NN	O	O
on	NN	O	O
a	NN	O	O
population	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
have	NN	O	O
also	NN	O	O
examined	NN	O	O
various	NN	O	O
parameters	NN	O	O
regulating	NN	O	O
the	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

Whereas	NN	O	O
IL-4	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
repress	NN	O	O
functional	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
through	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-12R	NN	O	B-protein
beta	NN	O	I-protein
2	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
,	NN	O	O
and	NN	O	O
IFN-alpha	NN	O	B-protein
in	NN	O	O
the	NN	O	O
human	NN	O	O
appear	NN	O	O
to	NN	O	O
induce	NN	O	O
IL-12R	NN	O	B-protein
beta	NN	O	I-protein
2	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
promote	NN	O	O
IL-12	NN	O	B-protein
responsiveness	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
Th1	NN	O	O
development	NN	O	O
can	NN	O	O
be	NN	O	O
considered	NN	O	O
in	NN	O	O
two	NN	O	O
stages	NN	O	O
,	NN	O	O
capacitance	NN	O	O
and	NN	O	O
development	NN	O	O
.	NN	O	O

Capacitance	NN	O	O
would	NN	O	O
simply	NN	O	O
involve	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-12R	NN	O	B-protein
beta	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	I-protein
beta	NN	O	I-protein
2	NN	O	I-protein
subunits	NN	O	I-protein
,	NN	O	O
regulated	NN	O	O
by	NN	O	O
TCR	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IFNs	NN	O	B-protein
.	NN	O	O

The	NN	O	O
second	NN	O	O
stage	NN	O	O
,	NN	O	O
development	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
is	NN	O	O
the	NN	O	O
true	NN	O	O
IL-12	NN	O	B-protein
induced	NN	O	O
developmental	NN	O	O
stage	NN	O	O
,	NN	O	O
involving	NN	O	O
expression	NN	O	O
of	NN	O	O
Stat4	NN	O	B-protein
inducible	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
human	NN	O	O
,	NN	O	O
this	NN	O	O
may	NN	O	O
also	NN	O	O
occur	NN	O	O
via	NN	O	O
IFN-alpha	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
activate	NN	O	O
Stat4	NN	O	B-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
perhaps	NN	O	O
possible	NN	O	O
that	NN	O	O
all	NN	O	O
of	NN	O	O
Stat4	NN	O	B-protein
actions	NN	O	O
on	NN	O	O
Th1	NN	O	O
development	NN	O	O
may	NN	O	O
be	NN	O	O
exert	NN	O	O
directly	NN	O	O
by	NN	O	O
Stat4	NN	O	B-protein
at	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
however	NN	O	O
we	NN	O	O
suggest	NN	O	O
that	NN	O	O
,	NN	O	O
more	NN	O	O
likely	NN	O	O
,	NN	O	O
Stat4	NN	O	B-protein
may	NN	O	O
act	NN	O	O
to	NN	O	O
induce	NN	O	O
Th1	NN	O	O
development	NN	O	O
through	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
other	NN	O	O
non-cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
whose	NN	O	O
stable	NN	O	O
expression	NN	O	O
maintains	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
state	NN	O	O
of	NN	O	O
a	NN	O	O
Th1	NN	O	B-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Lineage-specific	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
by	NN	O	O
interferon-gamma	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
to	NN	O	O
its	NN	O	O
heterodimeric	NN	O	O
receptor	NN	O	O
induces	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
JAK1	NN	O	B-protein
and	NN	O	O
JAK2	NN	O	B-protein
followed	NN	O	O
by	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1alpha	NN	O	B-protein
.	NN	O	O

Selective	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT1alpha	NN	O	B-protein
at	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
achieved	NN	O	O
by	NN	O	O
specific	NN	O	O
interaction	NN	O	O
between	NN	O	O
a	NN	O	O
cytosolic	NN	O	B-protein
tyrosine	NN	O	I-protein
motif	NN	O	I-protein
including	NN	O	O
Y440	NN	O	B-protein
in	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
receptor	NN	O	I-protein
alpha-chain	NN	O	I-protein
and	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
STAT1alpha	NN	O	B-protein
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
STAT1alpha	NN	O	B-protein
,	NN	O	O
STAT3	NN	O	B-protein
is	NN	O	O
also	NN	O	O
activated	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
was	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
,	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
the	NN	O	O
STAT3	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
by	NN	O	O
IFN-gamma	NN	O	B-protein
was	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
that	NN	O	O
are	NN	O	O
differentiated	NN	O	O
in	NN	O	O
vitro	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
CD34+	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
a	NN	O	O
single	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
can	NN	O	O
activate	NN	O	O
different	NN	O	O
STAT	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
in	NN	O	O
a	NN	O	O
cell-specific	NN	O	O
manner	NN	O	O
,	NN	O	O
which	NN	O	O
might	NN	O	O
result	NN	O	O
in	NN	O	O
cell-specific	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Alternative	NN	O	O
polyadenylation	NN	O	O
events	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-ATc	NN	O	B-protein
in	NN	O	O
effector	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-ATc	NN	O	B-protein
is	NN	O	O
synthesized	NN	O	O
in	NN	O	O
three	NN	O	O
prominent	NN	O	O
isoforms	NN	O	B-protein
.	NN	O	O

These	NN	O	O
differ	NN	O	O
in	NN	O	O
the	NN	O	O
length	NN	O	O
of	NN	O	O
their	NN	O	O
C	NN	O	O
terminal	NN	O	O
peptides	NN	O	O
and	NN	O	O
mode	NN	O	O
of	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Due	NN	O	O
to	NN	O	O
a	NN	O	O
switch	NN	O	O
from	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
polyA	NN	O	I-DNA
site	NN	O	I-DNA
to	NN	O	O
a	NN	O	O
more	NN	O	O
proximal	NN	O	B-DNA
polyA	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
NF-ATc	NN	O	B-protein
expression	NN	O	O
switches	NN	O	O
from	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
longer	NN	O	O
isoforms	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
that	NN	O	O
of	NN	O	O
short	NN	O	O
isoform	NN	O	B-protein
A	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
effector	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
relative	NN	O	O
low	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
cleavage	NN	O	B-protein
stimulation	NN	O	I-protein
factor	NN	O	I-protein
CstF-64	NN	O	B-protein
to	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
polyA	NN	O	I-DNA
site	NN	O	I-DNA
seems	NN	O	O
to	NN	O	O
contribute	NN	O	O
to	NN	O	O
its	NN	O	O
neglect	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
alternative	NN	O	O
polyadenylation	NN	O	O
events	NN	O	O
ensure	NN	O	O
the	NN	O	O
rapid	NN	O	O
accumulation	NN	O	O
of	NN	O	O
high	NN	O	O
concentrations	NN	O	O
of	NN	O	O
NF-ATc	NN	O	B-protein
necessary	NN	O	O
to	NN	O	O
exceed	NN	O	O
critical	NN	O	O
threshold	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-ATc	NN	O	B-protein
for	NN	O	O
gene	NN	O	O
induction	NN	O	O
in	NN	O	O
effector	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Estrogen	NN	O	O
and	NN	O	O
progesterone	NN	O	O
induction	NN	O	O
of	NN	O	O
survival	NN	O	O
of	NN	O	O
monoblastoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
undergoing	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
mononucleated	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
a	NN	O	O
physiological	NN	O	O
process	NN	O	O
for	NN	O	O
regulating	NN	O	O
the	NN	O	O
intensity	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
female	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
estrogen	NN	O	O
(	NN	O	O
E2	NN	O	O
)	NN	O	O
and	NN	O	O
progesterone	NN	O	O
(	NN	O	O
Prog	NN	O	O
)	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
modulate	NN	O	O
the	NN	O	O
reactivity	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
;	NN	O	O
recently	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
they	NN	O	O
can	NN	O	O
regulate	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
osteoblasts	NN	O	B-cell_type
.	NN	O	O

TNF-alpha	NN	O	B-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
has	NN	O	O
been	NN	O	O
well	NN	O	O
characterized	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
whether	NN	O	O
E2	NN	O	O
and	NN	O	O
Prog	NN	O	O
could	NN	O	O
interfere	NN	O	O
with	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
the	NN	O	O
monoblastoid	NN	O	B-cell_line
U937	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
E2	NN	O	O
or	NN	O	O
Prog	NN	O	O
increased	NN	O	O
survival	NN	O	O
and	NN	O	O
prevented	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
TNF-alpha	NN	O	B-protein
in	NN	O	O
both	NN	O	O
undifferentiated	NN	O	O
and	NN	O	O
macrophage-like	NN	O	O
PMA-differentiated	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
trypan	NN	O	O
blue	NN	O	O
exclusion	NN	O	O
cell	NN	O	O
counting	NN	O	O
,	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
,	NN	O	O
AnnexinV	NN	O	O
labeling	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
and	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
studies	NN	O	O
.	NN	O	O

This	NN	O	O
effect	NN	O	O
can	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
specific	NN	O	O
hormone	NN	O	O
receptors	NN	O	O
,	NN	O	O
since	NN	O	O
we	NN	O	O
observed	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
estrogen	NN	O	B-RNA
receptor	NN	O	I-RNA
alpha	NN	O	I-RNA
(	NN	O	I-RNA
ER-alpha	NN	O	I-RNA
)	NN	O	I-RNA
,	NN	O	I-RNA
ER-beta	NN	O	I-RNA
,	NN	O	I-RNA
and	NN	O	I-RNA
progesterone	NN	O	I-RNA
receptor	NN	O	I-RNA
(	NN	O	I-RNA
PR	NN	O	I-RNA
)	NN	O	I-RNA
mRNAs	NN	O	I-RNA
;	NN	O	O
the	NN	O	O
ER-alpha	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
immunocytochemical	NN	O	O
analysis	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
hormone-mediated	NN	O	O
survival	NN	O	O
against	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
concentration	NN	O	O
dependent	NN	O	O
,	NN	O	O
reaching	NN	O	O
the	NN	O	O
half-maximal	NN	O	O
effect	NN	O	O
at	NN	O	O
10	NN	O	O
nM	NN	O	O
and	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
ER	NN	O	O
antagonist	NN	O	O
ICI	NN	O	O
182	NN	O	O
,	NN	O	O
780	NN	O	O
in	NN	O	O
undifferentiated	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
further	NN	O	O
supporting	NN	O	O
a	NN	O	O
receptor-mediated	NN	O	O
mechanism	NN	O	O
of	NN	O	O
cell	NN	O	O
survival	NN	O	O
.	NN	O	O

Other	NN	O	O
steroid	NN	O	O
receptor	NN	O	O
drugs	NN	O	O
such	NN	O	O
as	NN	O	O
Raloxifene	NN	O	O
,	NN	O	O
RU486	NN	O	O
,	NN	O	O
or	NN	O	O
the	NN	O	O
ICI	NN	O	O
182	NN	O	O
,	NN	O	O
780	NN	O	O
in	NN	O	O
PMA-differentiated	NN	O	B-cell_line
cells	NN	O	I-cell_line
displayed	NN	O	O
agonist	NN	O	O
activity	NN	O	O
by	NN	O	O
preventing	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
as	NN	O	O
efficiently	NN	O	O
as	NN	O	O
the	NN	O	O
hormones	NN	O	O
alone	NN	O	O
,	NN	O	O
providing	NN	O	O
further	NN	O	O
evidence	NN	O	O
to	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
steroid	NN	O	O
receptor	NN	O	O
drugs	NN	O	O
may	NN	O	O
manifest	NN	O	O
agonist	NN	O	O
or	NN	O	O
antagonist	NN	O	O
activities	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
cellular	NN	O	O
context	NN	O	O
in	NN	O	O
which	NN	O	O
they	NN	O	O
are	NN	O	O
studied	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
E2	NN	O	O
was	NN	O	O
also	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
time-dependent	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
proapoptotic	NN	O	B-protein
Nip-2	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
supporting	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
hormone	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
target	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
ER	NN	O	B-protein
and	NN	O	O
PR	NN	O	B-protein
can	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
endogenous	NN	O	O
or	NN	O	O
exogenous	NN	O	O
ligands	NN	O	O
to	NN	O	O
induce	NN	O	O
a	NN	O	O
genetic	NN	O	O
response	NN	O	O
that	NN	O	O
impairs	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
data	NN	O	O
presented	NN	O	O
here	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
female	NN	O	B-protein
steroid	NN	O	I-protein
receptors	NN	O	I-protein
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
by	NN	O	O
preventing	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
monoblastoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
this	NN	O	O
effect	NN	O	O
might	NN	O	O
have	NN	O	O
important	NN	O	O
consequences	NN	O	O
in	NN	O	O
the	NN	O	O
clinical	NN	O	O
use	NN	O	O
of	NN	O	O
steroid	NN	O	O
receptor	NN	O	O
drugs	NN	O	O
.	NN	O	O

--	NN	O	O
Vegeto	NN	O	O
,	NN	O	O
E.	NN	O	O
,	NN	O	O
Pollio	NN	O	O
,	NN	O	O
G.	NN	O	O
,	NN	O	O
Pellicciari	NN	O	O
,	NN	O	O
C.	NN	O	O
,	NN	O	O
Maggi	NN	O	O
,	NN	O	O
A	NN	O	O
.	NN	O	O

Estrogen	NN	O	O
and	NN	O	O
progesterone	NN	O	O
induction	NN	O	O
of	NN	O	O
survival	NN	O	O
of	NN	O	O
monoblastoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
undergoing	NN	O	O
TNF-alpha	NN	O	B-protein
-inuced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Dicarba-closo-dodecaboranes	NN	O	O
as	NN	O	O
a	NN	O	O
pharmacophore	NN	O	O
.	NN	O	O

Retinoidal	NN	O	O
antagonists	NN	O	O
and	NN	O	O
potential	NN	O	O
agonists	NN	O	O
.	NN	O	O

Synthesis	NN	O	O
and	NN	O	O
biological	NN	O	O
evaluation	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
dicarba-closo-dodecaborane	NN	O	O
(	NN	O	O
carborane	NN	O	O
)	NN	O	O
derivatives	NN	O	O
of	NN	O	O
retinoids	NN	O	O
are	NN	O	O
described	NN	O	O
.	NN	O	O

Their	NN	O	O
retinoidal	NN	O	O
activity	NN	O	O
were	NN	O	O
examined	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
the	NN	O	O
differentiation-inducing	NN	O	O
ability	NN	O	O
toward	NN	O	O
human	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

High	NN	O	O
retinoidal	NN	O	O
activity	NN	O	O
(	NN	O	O
agonist	NN	O	O
or	NN	O	O
antagonist	NN	O	O
for	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
)	NN	O	O
requires	NN	O	O
a	NN	O	O
carboxylic	NN	O	O
acid	NN	O	O
moiety	NN	O	O
and	NN	O	O
an	NN	O	O
appropriate	NN	O	O
hydrophobic	NN	O	O
group	NN	O	O
located	NN	O	O
at	NN	O	O
a	NN	O	O
suitable	NN	O	O
position	NN	O	O
on	NN	O	O
the	NN	O	O
molecule	NN	O	O
.	NN	O	O

The	NN	O	O
4-carboranyl-substituted	NN	O	O
compounds	NN	O	O
(	NN	O	O
7	NN	O	O
,	NN	O	O
11	NN	O	O
)	NN	O	O
showed	NN	O	O
antagonistic	NN	O	O
activity	NN	O	O
but	NN	O	O
no	NN	O	O
agonistic	NN	O	O
activity	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
potent	NN	O	O
synergist	NN	O	O
HX630	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
the	NN	O	O
3-carboranyl-substituted	NN	O	O
compounds	NN	O	O
(	NN	O	O
8	NN	O	O
,	NN	O	O
12	NN	O	O
)	NN	O	O
showed	NN	O	O
potential	NN	O	O
agonistic	NN	O	O
activity	NN	O	O
,	NN	O	O
but	NN	O	O
no	NN	O	O
antagonistic	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicates	NN	O	O
that	NN	O	O
carboranes	NN	O	O
are	NN	O	O
applicable	NN	O	O
as	NN	O	O
the	NN	O	O
hydrophobic	NN	O	O
moiety	NN	O	O
of	NN	O	O
biologically	NN	O	O
active	NN	O	O
molecules	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
signaling	NN	O	O
by	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase-targeted	NN	O	I-protein
hematopoietic	NN	O	I-protein
tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
(	NN	O	O
HePTP	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
to	NN	O	O
produce	NN	O	O
cytokines	NN	O	B-protein
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
counterbalance	NN	O	O
of	NN	O	O
protein-tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
and	NN	O	O
protein-tyrosine	NN	O	O
phosphatases	NN	O	O
,	NN	O	O
many	NN	O	O
of	NN	O	O
which	NN	O	O
have	NN	O	O
a	NN	O	O
high	NN	O	O
degree	NN	O	O
of	NN	O	O
substrate	NN	O	O
specificity	NN	O	O
because	NN	O	O
of	NN	O	O
physical	NN	O	O
association	NN	O	O
with	NN	O	O
their	NN	O	O
targets	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-protein
protein-tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
(	NN	O	O
HePTP	NN	O	B-protein
)	NN	O	O
results	NN	O	O
in	NN	O	O
suppression	NN	O	O
of	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
promoter	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Efforts	NN	O	O
to	NN	O	O
pinpoint	NN	O	O
the	NN	O	O
exact	NN	O	O
site	NN	O	O
of	NN	O	O
action	NN	O	O
and	NN	O	O
specificity	NN	O	O
of	NN	O	O
HePTP	NN	O	B-protein
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
cascade	NN	O	O
revealed	NN	O	O
that	NN	O	O
HePTP	NN	O	B-protein
acts	NN	O	O
directly	NN	O	O
on	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MAP	NN	O	I-protein
)	NN	O	I-protein
kinases	NN	O	I-protein
Erk1	NN	O	B-protein
and	NN	O	I-protein
2	NN	O	I-protein
and	NN	O	O
consequently	NN	O	O
reduces	NN	O	O
the	NN	O	O
magnitude	NN	O	O
and	NN	O	O
duration	NN	O	O
of	NN	O	O
their	NN	O	O
catalytic	NN	O	O
activation	NN	O	O
in	NN	O	O
intact	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
HePTP	NN	O	B-protein
had	NN	O	O
no	NN	O	O
effects	NN	O	O
on	NN	O	O
N-terminal	NN	O	B-protein
c-Jun	NN	O	I-protein
kinase	NN	O	I-protein
or	NN	O	O
on	NN	O	O
events	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
MAP	NN	O	O
kinases	NN	O	O
.	NN	O	O

The	NN	O	O
specificity	NN	O	O
of	NN	O	O
HePTP	NN	O	B-protein
correlated	NN	O	O
with	NN	O	O
its	NN	O	O
physical	NN	O	O
association	NN	O	O
through	NN	O	O
its	NN	O	O
noncatalytic	NN	O	O
N	NN	O	B-protein
terminus	NN	O	I-protein
with	NN	O	O
Erk	NN	O	B-protein
and	NN	O	O
another	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
p38	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Jnk	NN	O	B-protein
or	NN	O	O
other	NN	O	O
proteins	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
HePTP	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
negative	NN	O	O
role	NN	O	O
in	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
signaling	NN	O	O
by	NN	O	O
specifically	NN	O	O
regulating	NN	O	O
MAP	NN	O	B-protein
kinases	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
and	NN	O	O
at	NN	O	O
early	NN	O	O
time	NN	O	O
points	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
before	NN	O	O
the	NN	O	O
activation-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
dual-specific	NN	O	I-protein
MAP	NN	O	I-protein
kinase	NN	O	I-protein
phosphatases	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
mediates	NN	O	O
signal	NN	O	O
integration	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
/CD28	NN	O	B-protein
costimulation	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
murine	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Optimal	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
requires	NN	O	O
two	NN	O	O
signals	NN	O	O
,	NN	O	O
one	NN	O	O
generated	NN	O	O
by	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
another	NN	O	O
by	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
costimulatory	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
costimulation-induced	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
mouse	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
that	NN	O	O
reported	NN	O	O
for	NN	O	O
human	NN	O	B-cell_type
Jurkat	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Jun	NN	O	B-protein
NH2-terminal	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
is	NN	O	O
weakly	NN	O	O
activated	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
either	NN	O	O
anti-CD3	NN	O	B-protein
or	NN	O	O
anti-CD28	NN	O	B-protein
in	NN	O	O
murine	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
and	NN	O	O
splenic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
is	NN	O	O
activated	NN	O	O
strongly	NN	O	O
and	NN	O	O
synergistically	NN	O	O
by	NN	O	O
either	NN	O	O
CD3/CD28	NN	O	B-protein
coligation	NN	O	O
or	NN	O	O
PMA/Ca2+	NN	O	O
ionophore	NN	O	O
stimulation	NN	O	O
,	NN	O	O
which	NN	O	O
mimics	NN	O	O
TCR	NN	O	B-protein
-	NN	O	O
CD3/CD28	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
correlates	NN	O	O
closely	NN	O	O
with	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
PMA-induced	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
.	NN	O	O

T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
production	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
induced	NN	O	O
by	NN	O	O
both	NN	O	O
CD3	NN	O	B-protein
and	NN	O	O
CD3	NN	O	B-protein
/CD28	NN	O	B-protein
ligation	NN	O	O
and	NN	O	O
the	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	I-protein
ATF-2	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
each	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
inhibitor	NN	O	O
SB203580	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
p38	NN	O	B-protein
MAPK	NN	O	I-protein
1	NN	O	O
)	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
signal	NN	O	O
integration	NN	O	O
during	NN	O	O
costimulation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
mouse	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
2	NN	O	O
)	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
augmentation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
3	NN	O	O
)	NN	O	O
regulates	NN	O	O
whether	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
enter	NN	O	O
a	NN	O	O
state	NN	O	O
of	NN	O	O
functional	NN	O	O
unresponsiveness	NN	O	O
.	NN	O	O

-DOCSTART-	O

Erythroid	NN	O	O
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
differentially	NN	O	O
regulated	NN	O	O
by	NN	O	O
erythropoietin	NN	O	B-protein
,	NN	O	O
haemin	NN	O	O
and	NN	O	O
delta-aminolaevulinic	NN	O	O
acid	NN	O	O
in	NN	O	O
UT-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Erythropoietin	NN	O	O
(	NN	O	O
Epo	NN	O	O
)	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
later	NN	O	O
stages	NN	O	O
of	NN	O	O
erythropoiesis	NN	O	O
,	NN	O	O
acting	NN	O	O
to	NN	O	O
promote	NN	O	O
cell	NN	O	O
survival	NN	O	O
and	NN	O	O
proliferation	NN	O	O
,	NN	O	O
but	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
differentiation	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
defined	NN	O	O
.	NN	O	O

The	NN	O	O
UT-7	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
exhibits	NN	O	O
both	NN	O	O
erythroid	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
characteristics	NN	O	O
and	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
along	NN	O	O
the	NN	O	O
erythroid	NN	O	O
pathway	NN	O	O
by	NN	O	O
Epo	NN	O	B-protein
or	NN	O	O
the	NN	O	O
megakaryocytic	NN	O	O
pathway	NN	O	O
by	NN	O	O
phorbol	NN	O	O
myristic	NN	O	O
acetate	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
compared	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
Epo	NN	O	B-protein
and	NN	O	O
the	NN	O	O
chemical	NN	O	O
inducers	NN	O	O
,	NN	O	O
delta-aminolaevulinic	NN	O	O
acid	NN	O	O
(	NN	O	O
delta-ALA	NN	O	O
)	NN	O	O
and	NN	O	O
haemin	NN	O	O
on	NN	O	O
the	NN	O	O
differentiation	NN	O	O
capacity	NN	O	O
of	NN	O	O
UT-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Epo	NN	O	B-protein
alone	NN	O	O
promoted	NN	O	O
relatively	NN	O	O
early	NN	O	O
events	NN	O	O
in	NN	O	O
erythroid	NN	O	O
maturation	NN	O	O
,	NN	O	O
without	NN	O	O
significant	NN	O	O
changes	NN	O	O
in	NN	O	O
haemoglobin	NN	O	O
production	NN	O	O
or	NN	O	O
morphology	NN	O	O
.	NN	O	O

GATA-2	NN	O	B-protein
and	NN	O	O
c-myb	NN	O	B-protein
were	NN	O	O
down-regulated	NN	O	O
by	NN	O	O
Epo	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GATA-2	NN	O	B-protein
was	NN	O	O
further	NN	O	O
down-modulated	NN	O	O
by	NN	O	O
the	NN	O	O
inducers	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
SCL	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
Epo	NN	O	B-protein
and	NN	O	O
further	NN	O	O
increased	NN	O	O
by	NN	O	O
haemin	NN	O	O
and	NN	O	O
delta-ALA	NN	O	O
.	NN	O	O

Epo	NN	O	B-protein
caused	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
cells	NN	O	O
expressing	NN	O	O
cell	NN	O	O
surface	NN	O	O
glycophorin	NN	O	B-protein
A	NN	O	I-protein
(	NN	O	O
GPA	NN	O	B-protein
)	NN	O	O
and	NN	O	O
up-regulated	NN	O	O
beta-	NN	O	B-protein
and	NN	O	I-protein
gamma-globin	NN	O	I-protein
by	NN	O	O
several	NN	O	O
fold	NN	O	O
.	NN	O	O

Both	NN	O	O
haemin	NN	O	O
and	NN	O	O
delta-ALA	NN	O	O
caused	NN	O	O
a	NN	O	O
de	NN	O	O
novo	NN	O	O
increase	NN	O	O
in	NN	O	O
alpha-globin	NN	O	B-protein
expression	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
enhancing	NN	O	O
Epo	NN	O	B-protein
-induced	NN	O	O
beta-globin	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
a	NN	O	O
marked	NN	O	O
increase	NN	O	O
in	NN	O	O
haemoglobin	NN	O	O
production	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
haemoglobin	NN	O	O
production	NN	O	O
in	NN	O	O
UT-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
limited	NN	O	O
by	NN	O	O
a	NN	O	O
deficiency	NN	O	O
of	NN	O	O
erythroid-specific	NN	O	B-protein
aminolaevulinic	NN	O	I-protein
acid	NN	O	I-protein
synthase	NN	O	I-protein
(	NN	O	O
ALAS-E	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
or	NN	O	O
globin	NN	O	O
synthesis	NN	O	O
as	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
their	NN	O	O
immaturity	NN	O	O
as	NN	O	O
a	NN	O	O
multipotential	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Essential	NN	O	O
role	NN	O	O
of	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
in	NN	O	O
intrapulmonary	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Acute	NN	O	O
inflammatory	NN	O	O
injury	NN	O	O
in	NN	O	O
rat	NN	O	O
lung	NN	O	O
induced	NN	O	O
by	NN	O	O
deposition	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-protein
G	NN	O	I-protein
immune	NN	O	I-protein
complexes	NN	O	I-protein
requires	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
chemokines	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappaB	NN	O	I-protein
.	NN	O	O

There	NN	O	O
is	NN	O	O
little	NN	O	O
direct	NN	O	O
evidence	NN	O	O
regarding	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
in	NN	O	O
these	NN	O	O
activation	NN	O	O
events	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
studies	NN	O	O
,	NN	O	O
rat	NN	O	O
lungs	NN	O	O
were	NN	O	O
depleted	NN	O	O
of	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
by	NN	O	O
airway	NN	O	O
instillation	NN	O	O
of	NN	O	O
liposome-encapsulated	NN	O	O
dichloromethylene	NN	O	O
diphosphonate	NN	O	O
.	NN	O	O

These	NN	O	O
procedures	NN	O	O
,	NN	O	O
which	NN	O	O
greatly	NN	O	O
reduced	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
retrievable	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
suppressed	NN	O	O
activation	NN	O	O
of	NN	O	O
lung	NN	O	B-protein
NF-kappaB	NN	O	I-protein
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
model	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
bronchoalveolar	NN	O	O
lavage	NN	O	O
levels	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
CXC	NN	O	B-protein
chemokine	NN	O	I-protein
,	NN	O	O
macrophage	NN	O	B-protein
inflammatory	NN	O	I-protein
protein-2	NN	O	I-protein
,	NN	O	O
were	NN	O	O
substantially	NN	O	O
reduced	NN	O	O
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
,	NN	O	O
upregulation	NN	O	O
of	NN	O	O
the	NN	O	O
lung	NN	O	B-protein
vascular	NN	O	I-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
,	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
,	NN	O	O
was	NN	O	O
greatly	NN	O	O
reduced	NN	O	O
by	NN	O	O
intrapulmonary	NN	O	O
instillation	NN	O	O
of	NN	O	O
phosphonate-containing	NN	O	O
liposomes	NN	O	O
.	NN	O	O

Neutrophil	NN	O	O
accumulation	NN	O	O
and	NN	O	O
development	NN	O	O
of	NN	O	O
lung	NN	O	O
injury	NN	O	O
were	NN	O	O
also	NN	O	O
substantially	NN	O	O
diminished	NN	O	O
.	NN	O	O

Lung	NN	O	O
instillation	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
in	NN	O	O
alveolar	NN	O	O
macrophage-depleted	NN	O	O
rats	NN	O	O
restored	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
response	NN	O	O
in	NN	O	O
whole	NN	O	O
lung	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
this	NN	O	O
inflammatory	NN	O	O
model	NN	O	O
,	NN	O	O
initial	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
occurs	NN	O	O
in	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
ensuing	NN	O	O
production	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
may	NN	O	O
propagate	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
to	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
in	NN	O	O
the	NN	O	O
lung	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interferon-beta	NN	O	B-protein
mediates	NN	O	O
stromal	NN	O	O
cell	NN	O	O
rescue	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
resolution	NN	O	O
of	NN	O	O
immune	NN	O	O
responses	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
extensive	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
to	NN	O	O
generate	NN	O	O
and	NN	O	O
maintain	NN	O	O
immunological	NN	O	O
memory	NN	O	O
,	NN	O	O
some	NN	O	O
antigen-specific	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
must	NN	O	O
survive	NN	O	O
and	NN	O	O
revert	NN	O	O
to	NN	O	O
a	NN	O	O
resting	NN	O	O
G0/G1	NN	O	O
state	NN	O	O
.	NN	O	O

Cytokines	NN	O	B-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
common	NN	O	O
gamma	NN	O	B-protein
chain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	O
promote	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
blasts	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
also	NN	O	O
induce	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
soluble	NN	O	B-protein
factors	NN	O	I-protein
secreted	NN	O	O
by	NN	O	O
stromal	NN	O	B-cell_type
cells	NN	O	I-cell_type
induce	NN	O	O
Tcell	NN	O	O
survival	NN	O	O
in	NN	O	O
a	NN	O	O
resting	NN	O	O
G0/G1	NN	O	O
state	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
that	NN	O	O
interferon-beta	NN	O	O
is	NN	O	O
the	NN	O	O
principal	NN	O	O
mediator	NN	O	O
of	NN	O	O
stromal	NN	O	O
cell-mediated	NN	O	O
Tcell	NN	O	O
rescue	NN	O	O
from	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Interferon-alpha	NN	O	B-protein
and	NN	O	I-protein
-beta	NN	O	I-protein
promote	NN	O	O
the	NN	O	O
reversion	NN	O	O
of	NN	O	O
blast	NN	O	B-cell_type
Tcells	NN	O	I-cell_type
to	NN	O	O
a	NN	O	O
resting	NN	O	O
G0/G1	NN	O	O
configuration	NN	O	O
with	NN	O	O
all	NN	O	O
the	NN	O	O
characteristic	NN	O	O
features	NN	O	O
of	NN	O	O
stromal	NN	O	O
cell	NN	O	O
rescue	NN	O	O
;	NN	O	O
such	NN	O	O
as	NN	O	O
high	NN	O	O
Bcl-XL	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
low	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

Type	NN	O	B-protein
I	NN	O	I-protein
interferons	NN	O	I-protein
and	NN	O	O
stromal	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulate	NN	O	O
apparently	NN	O	O
identical	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
STAT-1	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
mechanism	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
fundamental	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
persistence	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
sites	NN	O	O
of	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
;	NN	O	O
suggesting	NN	O	O
that	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
is	NN	O	O
an	NN	O	O
aberrant	NN	O	O
consequence	NN	O	O
of	NN	O	O
immunological	NN	O	O
memory	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reduction	NN	O	O
of	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
signalling	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
thalassaemia	NN	O	O
or	NN	O	O
sickle	NN	O	O
cell	NN	O	O
anaemia	NN	O	O
upon	NN	O	O
treatment	NN	O	O
with	NN	O	O
desferrioxamine	NN	O	O
.	NN	O	O

Recent	NN	O	O
evidence	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
progression	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
disease	NN	O	O
is	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
in	NN	O	O
thalassaemia	NN	O	O
major	NN	O	O
patients	NN	O	O
upon	NN	O	O
treatment	NN	O	O
with	NN	O	O
high	NN	O	O
doses	NN	O	O
of	NN	O	O
desferrioxamine	NN	O	O
(	NN	O	O
DFX	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
authors	NN	O	O
have	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
in	NN	O	O
vitro	NN	O	O
exposure	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
DFX	NN	O	O
decreases	NN	O	O
the	NN	O	O
bioavailability	NN	O	O
of	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
which	NN	O	O
has	NN	O	O
a	NN	O	O
stimulatory	NN	O	O
effect	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-protein
bioavailability	NN	O	O
from	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
10	NN	O	O
patients	NN	O	O
with	NN	O	O
thalassaemia	NN	O	O
or	NN	O	O
sickle	NN	O	O
cell	NN	O	O
anaemia	NN	O	O
given	NN	O	O
DFX	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
10	NN	O	O
untreated	NN	O	O
subjects	NN	O	O
has	NN	O	O
been	NN	O	O
evaluated	NN	O	O
.	NN	O	O

Evidence	NN	O	O
is	NN	O	O
presented	NN	O	O
showing	NN	O	O
that	NN	O	O
DFX	NN	O	O
treatment	NN	O	O
reduces	NN	O	O
TNF-alpha	NN	O	B-protein
bioavailability	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
its	NN	O	O
steady	NN	O	O
state	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
and	NN	O	O
by	NN	O	O
enhancing	NN	O	O
its	NN	O	O
inactivation	NN	O	O
through	NN	O	O
binding	NN	O	O
to	NN	O	O
soluble	NN	O	O
TNF-alpha	NN	O	B-protein
receptor	NN	O	O
type	NN	O	O
II	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
DFX	NN	O	O
treatment	NN	O	O
limits	NN	O	O
the	NN	O	O
in	NN	O	O
vivo	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
both	NN	O	O
TNF-alpha	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
signalling	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.005	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
TNF-alpha	NN	O	B-protein
bioavailability	NN	O	O
and	NN	O	O
signalling	NN	O	O
are	NN	O	O
impaired	NN	O	O
in	NN	O	O
patients	NN	O	O
upon	NN	O	O
DFX	NN	O	O
treatment	NN	O	O
.	NN	O	O

This	NN	O	O
mechanism	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
delayed	NN	O	O
progression	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
p50/p65	NN	O	I-protein
heterodimer	NN	O	I-protein
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
pro-interleukin-1beta	NN	O	I-DNA
gene	NN	O	I-DNA
at	NN	O	O
two	NN	O	O
subregions	NN	O	O
of	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
region	NN	O	O
between-3134	NN	O	O
and	NN	O	O
-2729	NN	O	O
bp	NN	O	O
upstream	NN	O	O
from	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
pro-interleukin	NN	O	I-DNA
1beta	NN	O	I-DNA
(	NN	O	I-DNA
proIL-1beta	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
identified	NN	O	O
as	NN	O	O
an	NN	O	O
LPS-responsive	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
the	NN	O	O
sequences	NN	O	O
located	NN	O	O
between	NN	O	B-DNA
-3134	NN	O	I-DNA
and	NN	O	I-DNA
-2987	NN	O	I-DNA
on	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
proIL-1beta	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
LPS-stimulated	NN	O	B-cell_line
Raw	NN	O	I-cell_line
264.7	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
examined	NN	O	O
in	NN	O	O
detail	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
obtained	NN	O	O
by	NN	O	O
transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
fos	NN	O	O
-CAT	NN	O	B-DNA
constructs	NN	O	I-DNA
that	NN	O	O
contained	NN	O	O
serial	NN	O	B-DNA
5'-deletion	NN	O	I-DNA
mutations	NN	O	I-DNA
showed	NN	O	O
that	NN	O	O
the	NN	O	O
region	NN	O	O
between	NN	O	O
-3134	NN	O	O
and	NN	O	O
-3059	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
transcription	NN	O	O
by	NN	O	O
LPS	NN	O	O
.	NN	O	O

Gel	NN	O	O
shift	NN	O	O
assay	NN	O	O
studies	NN	O	O
with	NN	O	O
synthetic	NN	O	B-DNA
oligonucleotides	NN	O	I-DNA
corresponding	NN	O	O
to	NN	O	O
partial	NN	O	O
sequences	NN	O	O
of	NN	O	O
the	NN	O	O
latter	NN	O	O
region	NN	O	O
and	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
stimulated	NN	O	O
cells	NN	O	O
revealed	NN	O	O
specific	NN	O	O
protein	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
between	NN	O	O
-3110	NN	O	O
and	NN	O	O
-3090	NN	O	O
and	NN	O	O
between	NN	O	O
-3079	NN	O	O
and	NN	O	O
-3059	NN	O	O
.	NN	O	O

These	NN	O	O
specific	NN	O	O
bindings	NN	O	O
were	NN	O	O
time	NN	O	O
and	NN	O	O
LPS	NN	O	O
dose	NN	O	O
dependent	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
supershift	NN	O	O
analysis	NN	O	O
using	NN	O	O
specific	NN	O	O
antibodies	NN	O	O
against	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
suggested	NN	O	O
that	NN	O	O
both	NN	O	O
binding	NN	O	O
complexes	NN	O	O
contained	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
components	NN	O	I-protein
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
did	NN	O	O
not	NN	O	O
contain	NN	O	O
other	NN	O	O
NF-kappaB	NN	O	B-protein
proteins	NN	O	I-protein
(	NN	O	O
p52	NN	O	B-protein
,	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
Rel	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
AP-1	NN	O	B-protein
proteins	NN	O	I-protein
(	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
C-Jun	NN	O	B-protein
)	NN	O	O
,	NN	O	O
CREB	NN	O	B-protein
or	NN	O	O
C/EBPbeta	NN	O	B-protein
(	NN	O	O
NF-IL6	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
either	NN	O	O
of	NN	O	O
the	NN	O	O
putative	NN	O	O
NF-kappaB-binding	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
enhancer	NN	O	O
element	NN	O	O
decreased	NN	O	O
the	NN	O	O
LPS-stimulated	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicated	NN	O	O
that	NN	O	O
two	NN	O	O
NF-kappaB-binding	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
are	NN	O	O
located	NN	O	O
between	NN	O	O
-3134	NN	O	B-DNA
and	NN	O	I-DNA
-3059	NN	O	I-DNA
,	NN	O	O
are	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
proIL-1beta	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

N-acetyl-L-cysteine	NN	O	O
inhibits	NN	O	O
primary	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
at	NN	O	O
the	NN	O	O
dendritic	NN	O	O
cell	NN	O	O
level	NN	O	O
:	NN	O	O
association	NN	O	O
with	NN	O	O
NF-kappaB	NN	O	B-protein
inhibition	NN	O	O
.	NN	O	O

N-acetyl-L-cysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
antioxidant	NN	O	O
molecule	NN	O	O
endowed	NN	O	O
with	NN	O	O
immunomodulatory	NN	O	O
properties	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
NAC	NN	O	O
on	NN	O	O
the	NN	O	O
induction	NN	O	O
phase	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
its	NN	O	O
action	NN	O	O
on	NN	O	O
human	NN	O	O
dendritic	NN	O	O
cells	NN	O	O
(	NN	O	O
DC	NN	O	O
)	NN	O	O
derived	NN	O	O
from	NN	O	O
adherent	NN	O	O
PBMC	NN	O	O
cultured	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
granulocyte-macrophage	NN	O	B-protein
CSF	NN	O	I-protein
.	NN	O	O

We	NN	O	O
first	NN	O	O
found	NN	O	O
that	NN	O	O
NAC	NN	O	O
inhibited	NN	O	O
the	NN	O	O
constitutive	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
LPS-induced	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
,	NN	O	O
NAC	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
down-regulate	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
by	NN	O	O
DC	NN	O	B-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
their	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
,	NN	O	O
CD86	NN	O	B-protein
(	NN	O	O
B7-2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
CD40	NN	O	B-protein
molecules	NN	O	I-protein
both	NN	O	O
at	NN	O	O
the	NN	O	O
basal	NN	O	O
state	NN	O	O
and	NN	O	O
upon	NN	O	O
LPS	NN	O	O
activation	NN	O	O
.	NN	O	O

NAC	NN	O	O
also	NN	O	O
inhibited	NN	O	O
DC	NN	O	O
responses	NN	O	O
induced	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
engagement	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
NAC	NN	O	O
were	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
nonspecific	NN	O	O
toxicity	NN	O	O
as	NN	O	O
neither	NN	O	O
the	NN	O	O
viability	NN	O	O
of	NN	O	O
DC	NN	O	B-cell_type
nor	NN	O	O
their	NN	O	O
mannose	NN	O	O
receptor-mediated	NN	O	O
endocytosis	NN	O	O
were	NN	O	O
modified	NN	O	O
by	NN	O	O
NAC	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
NAC	NN	O	O
to	NN	O	O
MLR	NN	O	O
between	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
allogeneic	NN	O	O
DC	NN	O	B-cell_type
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
profound	NN	O	O
inhibition	NN	O	O
of	NN	O	O
alloreactive	NN	O	O
responses	NN	O	O
,	NN	O	O
which	NN	O	O
could	NN	O	O
be	NN	O	O
attributed	NN	O	O
to	NN	O	O
a	NN	O	O
defect	NN	O	O
of	NN	O	O
DC	NN	O	B-cell_type
as	NN	O	O
APC-independent	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
were	NN	O	O
not	NN	O	O
inhibited	NN	O	O
by	NN	O	O
NAC	NN	O	O
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
NAC	NN	O	O
might	NN	O	O
impair	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
primary	NN	O	O
immune	NN	O	O
responses	NN	O	O
in	NN	O	O
humans	NN	O	O
through	NN	O	O
its	NN	O	O
inhibitory	NN	O	O
action	NN	O	O
on	NN	O	O
DC	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Thrombin-induced	NN	O	B-protein
p65	NN	O	I-protein
homodimer	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
downstream	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
mediates	NN	O	O
endothelial	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
neutrophil	NN	O	B-cell_type
adhesion	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
proinflammatory	NN	O	B-protein
mediator	NN	O	I-protein
,	NN	O	O
thrombin	NN	O	B-protein
,	NN	O	O
released	NN	O	O
during	NN	O	O
intravascular	NN	O	O
coagulation	NN	O	O
and	NN	O	O
tissue	NN	O	O
injury	NN	O	O
,	NN	O	O
induces	NN	O	O
ICAM-1	NN	O	B-protein
(	NN	O	O
CD54	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
HUVEC	NN	O	B-cell_line
with	NN	O	O
thrombin	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
dose-	NN	O	O
and	NN	O	O
time-dependent	NN	O	O
increases	NN	O	O
in	NN	O	O
ICAM-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
and	NN	O	O
in	NN	O	O
ICAM-1	NN	O	B-protein
-dependent	NN	O	O
endothelial	NN	O	O
adhesivity	NN	O	O
toward	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
ICAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
luciferase	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	I-DNA
ICAM-1LUC	NN	O	I-DNA
)	NN	O	I-DNA
constructs	NN	O	I-DNA
indicated	NN	O	O
that	NN	O	O
deletion	NN	O	O
of	NN	O	O
upstream	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
-533	NN	O	B-DNA
bases	NN	O	I-DNA
from	NN	O	O
translation	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
)	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
thrombin	NN	O	B-protein
responsiveness	NN	O	O
,	NN	O	O
whereas	NN	O	O
mutation/deletion	NN	O	O
of	NN	O	O
downstream	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
-223	NN	O	B-DNA
bases	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
translation	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
)	NN	O	O
prevented	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
downstream	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
is	NN	O	O
critical	NN	O	O
for	NN	O	O
thrombin	NN	O	B-protein
inducibility	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-directed	NN	O	O
luciferase	NN	O	B-protein
activity	NN	O	O
increased	NN	O	O
approximately	NN	O	O
3-fold	NN	O	O
when	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
plasmid	NN	O	B-DNA
pNF-kappa	NN	O	I-DNA
BLUC	NN	O	I-DNA
containing	NN	O	O
five	NN	O	O
copies	NN	O	O
of	NN	O	O
consensus	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
a	NN	O	O
minimal	NN	O	O
adenovirus	NN	O	B-DNA
E1B	NN	O	I-DNA
promoter-luciferase	NN	O	I-DNA
gene	NN	O	I-DNA
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
thrombin	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
essential	NN	O	O
for	NN	O	O
thrombin	NN	O	B-protein
response	NN	O	O
.	NN	O	O

Gel	NN	O	O
supershift	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
thrombin	NN	O	B-protein
induced	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
Bp65	NN	O	I-protein
(	NN	O	O
Rel	NN	O	B-protein
A	NN	O	I-protein
)	NN	O	O
to	NN	O	O
downstream	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Thrombin	NN	O	B-protein
receptor	NN	O	O
activation	NN	O	O
peptide	NN	O	O
,	NN	O	O
a	NN	O	O
14-amino-acid	NN	O	O
peptide	NN	O	O
representing	NN	O	O
the	NN	O	O
new	NN	O	O
NH2	NN	O	B-protein
terminus	NN	O	I-protein
of	NN	O	O
proteolytically	NN	O	O
activated	NN	O	O
receptor-1	NN	O	B-protein
,	NN	O	O
mimicked	NN	O	O
thrombin	NN	O	B-protein
's	NN	O	O
action	NN	O	O
in	NN	O	O
inducing	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
thrombin	NN	O	B-protein
activates	NN	O	O
endothelial	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocyte	NN	O	I-cell_type
adhesion	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
Bp65	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
downstream	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
ICAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
after	NN	O	O
proteolytically	NN	O	O
activated	NN	O	O
receptor-1	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Smad6	NN	O	B-protein
and	NN	O	O
Smad7	NN	O	B-protein
on	NN	O	O
bone	NN	O	O
morphogenetic	NN	O	O
protein-	NN	O	O
and	NN	O	O
activin-mediated	NN	O	O
growth	NN	O	O
arrest	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Smad6	NN	O	B-protein
and	NN	O	O
Smad7	NN	O	B-protein
prevent	NN	O	O
ligand-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
signal-transducing	NN	O	B-protein
Smad	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
the	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
both	NN	O	O
Smad6	NN	O	B-protein
and	NN	O	O
Smad7	NN	O	B-protein
are	NN	O	O
human	NN	O	B-protein
bone	NN	O	I-protein
morphogenetic	NN	O	I-protein
protein-2	NN	O	I-protein
(	NN	O	O
hBMP-2	NN	O	B-protein
)	NN	O	O
-inducible	NN	O	O
antagonists	NN	O	O
of	NN	O	O
hBMP-2	NN	O	B-protein
-induced	NN	O	O
growth	NN	O	O
arrest	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
mouse	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
hybridoma	NN	O	I-cell_line
HS-72	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
confirmed	NN	O	O
that	NN	O	O
the	NN	O	O
ectopic	NN	O	O
expressions	NN	O	O
of	NN	O	O
Smad6	NN	O	B-protein
and	NN	O	O
Smad7	NN	O	B-protein
inhibited	NN	O	O
the	NN	O	O
hBMP-2	NN	O	B-protein
-induced	NN	O	O
Smad1/Smad5	NN	O	B-protein
phosphorylation	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
Smad7	NN	O	B-protein
is	NN	O	O
an	NN	O	O
activin	NN	O	O
A-inducible	NN	O	O
antagonist	NN	O	O
of	NN	O	O
activin	NN	O	O
A-induced	NN	O	O
growth	NN	O	O
arrest	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
HS-72	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
although	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
Smad6	NN	O	B-protein
was	NN	O	O
induced	NN	O	O
by	NN	O	O
activin	NN	O	O
A	NN	O	O
in	NN	O	O
HS-72	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
Smad6	NN	O	B-protein
showed	NN	O	O
no	NN	O	O
antagonistic	NN	O	O
effect	NN	O	O
on	NN	O	O
activin	NN	O	O
A-induced	NN	O	O
growth	NN	O	O
arrest	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
Smad7	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Smad6	NN	O	B-protein
,	NN	O	O
inhibited	NN	O	O
the	NN	O	O
activin	NN	O	O
A-induced	NN	O	O
Smad2	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
HS-72	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Smad6	NN	O	B-protein
and	NN	O	O
Smad7	NN	O	B-protein
exhibit	NN	O	O
differential	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
in	NN	O	O
bone	NN	O	O
morphogenetic	NN	O	O
protein-2-	NN	O	O
and	NN	O	O
activin	NN	O	O
A-mediated	NN	O	O
signaling	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Retinoic	NN	O	O
acid	NN	O	O
induces	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
CD34+	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
involvement	NN	O	O
of	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
and	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptors	NN	O	I-protein
depends	NN	O	O
on	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
of	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Retinoids	NN	O	O
are	NN	O	O
bifunctional	NN	O	O
regulators	NN	O	O
of	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
explored	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
on	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
CD34+	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
normal	NN	O	O
bone	NN	O	O
marrow	NN	O	O
.	NN	O	O

RA	NN	O	O
(	NN	O	O
100	NN	O	O
nM	NN	O	O
)	NN	O	O
induced	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
dead	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
24	NN	O	O
%	NN	O	O
to	NN	O	O
44	NN	O	O
%	NN	O	O
at	NN	O	O
day	NN	O	O
6	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.05	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
6	NN	O	O
)	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
control	NN	O	B-cell_type
cells	NN	O	I-cell_type
cultured	NN	O	O
in	NN	O	O
medium	NN	O	O
alone	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
was	NN	O	O
dose	NN	O	O
dependent	NN	O	O
and	NN	O	O
appeared	NN	O	O
relatively	NN	O	O
late	NN	O	O
.	NN	O	O

Significant	NN	O	O
differences	NN	O	O
were	NN	O	O
observed	NN	O	O
from	NN	O	O
day	NN	O	O
4	NN	O	O
onward	NN	O	O
.	NN	O	O

Apoptosis	NN	O	O
,	NN	O	O
or	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
,	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
as	NN	O	O
the	NN	O	O
mode	NN	O	O
of	NN	O	O
cell	NN	O	O
death	NN	O	O
by	NN	O	O
using	NN	O	O
the	NN	O	O
TUNEL	NN	O	O
assay	NN	O	O
,	NN	O	O
which	NN	O	O
detects	NN	O	O
single	NN	O	O
strand	NN	O	O
nicks	NN	O	O
in	NN	O	O
DNA	NN	O	O
,	NN	O	O
or	NN	O	O
by	NN	O	O
the	NN	O	O
Nicoletti	NN	O	O
technique	NN	O	O
demonstrating	NN	O	O
a	NN	O	O
subdiploid	NN	O	O
population	NN	O	O
by	NN	O	O
DNA	NN	O	O
staining	NN	O	O
.	NN	O	O

RA	NN	O	O
previously	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
inhibit	NN	O	O
granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
--	NN	O	O
and	NN	O	O
not	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
--	NN	O	O
stimulated	NN	O	O
proliferation	NN	O	O
of	NN	O	O
CD34+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
RA	NN	O	O
opposed	NN	O	O
anti-apoptotic	NN	O	O
effects	NN	O	O
of	NN	O	O
G-CSF	NN	O	B-protein
and	NN	O	O
GM-CSF	NN	O	B-protein
on	NN	O	O
CD34+	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
G-CSF	NN	O	B-protein
:	NN	O	O
8	NN	O	O
%	NN	O	O
dead	NN	O	B-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
day	NN	O	O
6	NN	O	O
;	NN	O	O
G-CSF	NN	O	B-protein
+	NN	O	O
RA	NN	O	O
:	NN	O	O
20	NN	O	O
%	NN	O	O
;	NN	O	O
GM-CSF	NN	O	B-protein
:	NN	O	O
12	NN	O	O
%	NN	O	O
;	NN	O	O
GM-CSF	NN	O	B-protein
+	NN	O	O
RA	NN	O	O
:	NN	O	O
27	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
RA	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
CD34+	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
CD34+CD71+	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
erythropoietin	NN	O	B-protein
.	NN	O	O

To	NN	O	O
explore	NN	O	O
the	NN	O	O
receptor	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
involved	NN	O	O
in	NN	O	O
RA-induced	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
selective	NN	O	O
ligands	NN	O	O
for	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RARs	NN	O	O
;	NN	O	O
RO13-7410	NN	O	O
)	NN	O	O
and	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RXRs	NN	O	B-protein
;	NN	O	O
RO	NN	O	O
25-6603	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
RARs	NN	O	B-protein
were	NN	O	O
involved	NN	O	O
in	NN	O	O
RA-mediated	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
myeloid	NN	O	O
progenitor	NN	O	O
cells	NN	O	O
,	NN	O	O
whereas	NN	O	O
RARs	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
RXRs	NN	O	B-protein
were	NN	O	O
involved	NN	O	O
in	NN	O	O
RA-mediated	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Suppressive	NN	O	O
effects	NN	O	O
of	NN	O	O
anti-inflammatory	NN	O	O
agents	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Studies	NN	O	O
from	NN	O	O
our	NN	O	O
laboratory	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
earliest	NN	O	O
stages	NN	O	O
of	NN	O	O
atherosclerosis	NN	O	O
may	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
an	NN	O	O
autoimmune	NN	O	O
reaction	NN	O	O
against	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
protein	NN	O	I-protein
60	NN	O	I-protein
(	NN	O	O
Hsp60	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
interactions	NN	O	O
of	NN	O	O
Hsp60-specific	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
arterial	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
EC	NN	O	B-cell_type
)	NN	O	O
require	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
Hsp60	NN	O	B-protein
and	NN	O	O
certain	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
shown	NN	O	O
to	NN	O	O
be	NN	O	O
induced	NN	O	O
simultaneously	NN	O	O
in	NN	O	O
EC	NN	O	B-cell_type
by	NN	O	O
mechanical	NN	O	O
and	NN	O	O
other	NN	O	O
types	NN	O	O
of	NN	O	O
stress	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
shown	NN	O	O
that	NN	O	O
suppression	NN	O	O
of	NN	O	O
T	NN	O	O
cell-mediated	NN	O	O
immune	NN	O	O
responses	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CyA	NN	O	O
)	NN	O	O
enhanced	NN	O	O
atherosclerotic	NN	O	O
lesion	NN	O	O
formation	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
aspirin	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
lower	NN	O	O
the	NN	O	O
risk	NN	O	O
of	NN	O	O
myocardial	NN	O	O
infarction	NN	O	O
in	NN	O	O
men	NN	O	O
.	NN	O	O

These	NN	O	O
conflicting	NN	O	O
observations	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
different	NN	O	O
effects	NN	O	O
of	NN	O	O
anti-inflammatory	NN	O	O
agents	NN	O	O
on	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
and	NN	O	O
Hsp	NN	O	O
expression	NN	O	O
in	NN	O	O
EC	NN	O	B-cell_type
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

MATERIAL	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
CyA	NN	O	O
,	NN	O	O
aspirin	NN	O	O
,	NN	O	O
and	NN	O	O
indomethacin	NN	O	O
on	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
using	NN	O	O
a	NN	O	O
proliferation	NN	O	O
assay	NN	O	O
.	NN	O	O

To	NN	O	O
explore	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
,	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
MCP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
Hsp60	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HUVECs	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analyses	NN	O	O
were	NN	O	O
used	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
activation	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
and	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
factor-1	NN	O	I-protein
(	NN	O	O
HSF-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
were	NN	O	O
performed	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
With	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
indomethacin	NN	O	O
,	NN	O	O
the	NN	O	O
used	NN	O	O
immunosuppressive	NN	O	O
and	NN	O	O
anti-inflammatory	NN	O	O
agents	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
influenza	NN	O	B-protein
virus	NN	O	I-protein
antigen	NN	O	I-protein
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
CyA	NN	O	O
and	NN	O	O
indomethacin	NN	O	O
did	NN	O	O
not	NN	O	O
suppress	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
-induced	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
expression	NN	O	O
on	NN	O	O
HUVECs	NN	O	B-cell_type
,	NN	O	O
whereas	NN	O	O
aspirin	NN	O	O
had	NN	O	O
an	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
EC	NN	O	B-cell_type
.	NN	O	O

All	NN	O	O
agents	NN	O	O
tested	NN	O	O
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
Hsp60	NN	O	B-protein
6	NN	O	O
hr	NN	O	O
after	NN	O	O
application	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
aspirin	NN	O	O
and	NN	O	O
indomethacin	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
CyA	NN	O	O
,	NN	O	O
induced	NN	O	O
Hsp70	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
HUVECs	NN	O	B-cell_type
that	NN	O	O
correlated	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
HSF-1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Our	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
tested	NN	O	O
agents	NN	O	O
(	NN	O	O
except	NN	O	O
indomethacin	NN	O	O
)	NN	O	O
are	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	O
cell-mediated	NN	O	O
immune	NN	O	O
response	NN	O	O
,	NN	O	O
as	NN	O	O
expected	NN	O	O
,	NN	O	O
that	NN	O	O
aspirin	NN	O	O
is	NN	O	O
an	NN	O	O
effective	NN	O	O
suppressor	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
all	NN	O	O
three	NN	O	O
agents	NN	O	O
can	NN	O	O
induce	NN	O	O
Hsp60	NN	O	B-protein
in	NN	O	O
HUVECs	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
anti-atherogenic	NN	O	O
effect	NN	O	O
of	NN	O	O
aspirin	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
prevention	NN	O	O
of	NN	O	O
the	NN	O	O
adhesion	NN	O	O
of	NN	O	O
sensitized	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
stressed	NN	O	O
EC	NN	O	B-cell_type
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
that	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
atherosclerosis-promoting	NN	O	O
effect	NN	O	O
of	NN	O	O
CyA	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
its	NN	O	O
potential	NN	O	O
as	NN	O	O
an	NN	O	O
inducer	NN	O	O
of	NN	O	O
Hsp60	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
its	NN	O	O
inability	NN	O	O
to	NN	O	O
down-regulate	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
expression	NN	O	O
on	NN	O	O
EC	NN	O	B-cell_type
;	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
that	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
aspirin	NN	O	O
may	NN	O	O
result	NN	O	O
in	NN	O	O
decreased	NN	O	O
recruitment	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
into	NN	O	O
the	NN	O	O
arterial	NN	O	O
intima	NN	O	O
beneath	NN	O	O
stressed	NN	O	O
EC	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
modulation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
inhaled	NN	O	O
fluticasone	NN	O	O
propionate	NN	O	O
in	NN	O	O
bronchial	NN	O	O
mucosa	NN	O	O
and	NN	O	O
blood	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
subjects	NN	O	O
with	NN	O	O
mild	NN	O	O
asthma	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
In	NN	O	O
vivo	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
provides	NN	O	O
a	NN	O	O
means	NN	O	O
of	NN	O	O
modulating	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
targeted	NN	O	O
cells	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
We	NN	O	O
sought	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
in	NN	O	O
vivo	NN	O	O
modulation	NN	O	O
of	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
by	NN	O	O
fluticasone	NN	O	O
propionate	NN	O	O
(	NN	O	O
FP	NN	O	O
)	NN	O	O
in	NN	O	O
subjects	NN	O	O
with	NN	O	O
mild	NN	O	O
asthma	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Ten	NN	O	O
atopic	NN	O	O
asthmatic	NN	O	O
subjects	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
FP	NN	O	O
250	NN	O	O
microg	NN	O	O
twice	NN	O	O
daily	NN	O	O
for	NN	O	O
4	NN	O	O
weeks	NN	O	O
.	NN	O	O

Before	NN	O	O
and	NN	O	O
after	NN	O	O
treatment	NN	O	O
,	NN	O	O
the	NN	O	O
patients	NN	O	O
underwent	NN	O	O
fiberoptic	NN	O	O
bronchoscopy	NN	O	O
with	NN	O	O
endobronchial	NN	O	O
biopsy	NN	O	O
and	NN	O	O
sampling	NN	O	O
of	NN	O	O
venous	NN	O	O
blood	NN	O	O
for	NN	O	O
measurements	NN	O	O
of	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

A	NN	O	O
solution	NN	O	O
hybridization	NN	O	O
assay	NN	O	O
was	NN	O	O
used	NN	O	O
for	NN	O	O
quantitative	NN	O	O
analysis	NN	O	O
of	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
a	NN	O	O
24-hour	NN	O	O
urinary	NN	O	O
cortisol	NN	O	O
excretion	NN	O	O
and	NN	O	O
an	NN	O	O
adrenocorticotropic	NN	O	O
hormone	NN	O	O
test	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
FP	NN	O	O
were	NN	O	O
performed	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
A	NN	O	O
high	NN	O	O
interindividual	NN	O	O
variation	NN	O	O
in	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
was	NN	O	O
seen	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
detected	NN	O	O
a	NN	O	O
significant	NN	O	O
reduction	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
in	NN	O	O
the	NN	O	O
endobronchial	NN	O	O
biopsy	NN	O	O
specimens	NN	O	O
after	NN	O	O
FP	NN	O	O
treatment	NN	O	O
(	NN	O	O
36.6	NN	O	O
+/-	NN	O	O
23.1	NN	O	O
and	NN	O	O
25.0	NN	O	O
+/-	NN	O	O
10.9	NN	O	O
amol	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
/microg	NN	O	O
RNA	NN	O	O
,	NN	O	O
respectively	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
.01	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
an	NN	O	O
even	NN	O	O
more	NN	O	O
striking	NN	O	O
downregulation	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	O
by	NN	O	O
its	NN	O	O
cognate	NN	O	O
ligand	NN	O	O
was	NN	O	O
documented	NN	O	O
(	NN	O	O
30.3	NN	O	O
+/-	NN	O	O
26.5	NN	O	O
and	NN	O	O
8.8	NN	O	O
+/-	NN	O	O
5	NN	O	O
amol	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
/microg	NN	O	O
RNA	NN	O	O
,	NN	O	O
respectively	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
.001	NN	O	O
)	NN	O	O
,	NN	O	O
possibly	NN	O	O
reflecting	NN	O	O
differences	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	O
sensitivity	NN	O	O
between	NN	O	O
tissues	NN	O	O
.	NN	O	O

A	NN	O	O
small	NN	O	O
but	NN	O	O
significant	NN	O	O
reduction	NN	O	O
of	NN	O	O
the	NN	O	O
24-hour	NN	O	O
urinary	NN	O	O
cortisol	NN	O	O
excretion	NN	O	O
was	NN	O	O
observed	NN	O	O
(	NN	O	O
233	NN	O	O
+/-	NN	O	O
109	NN	O	O
and	NN	O	O
157	NN	O	O
+/-	NN	O	O
66	NN	O	O
nmol/L	NN	O	O
,	NN	O	O
respectively	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
.01	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
feedback	NN	O	O
regulation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
synthesis	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
axis	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
the	NN	O	O
adrenocorticotropic	NN	O	O
hormone	NN	O	O
test	NN	O	O
remained	NN	O	O
normal	NN	O	O
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
FP	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
The	NN	O	O
results	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
confirm	NN	O	O
the	NN	O	O
potency	NN	O	O
of	NN	O	O
the	NN	O	O
inhaled	NN	O	O
corticosteroid	NN	O	O
FP	NN	O	O
and	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
considerable	NN	O	O
tissue-specific	NN	O	O
interindividual	NN	O	O
variation	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Corticoids	NN	O	O
and	NN	O	O
allergy	NN	O	O
]	NN	O	O

Inflammation	NN	O	O
is	NN	O	O
constantly	NN	O	O
observed	NN	O	O
in	NN	O	O
allergic	NN	O	O
reactions	NN	O	O
.	NN	O	O

Corticosteroids	NN	O	O
are	NN	O	O
most	NN	O	O
effective	NN	O	O
in	NN	O	O
preventing	NN	O	O
the	NN	O	O
late	NN	O	O
phase	NN	O	O
of	NN	O	O
allergic	NN	O	O
reaction	NN	O	O
.	NN	O	O

The	NN	O	O
action	NN	O	O
of	NN	O	O
glucocorticosteroids	NN	O	O
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
present	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

When	NN	O	O
activated	NN	O	O
,	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
form	NN	O	O
a	NN	O	O
dimer	NN	O	B-protein
and	NN	O	O
bind	NN	O	O
to	NN	O	O
DNA	NN	O	O
after	NN	O	O
migration	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Interaction	NN	O	O
to	NN	O	O
DNA	NN	O	O
induces	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
rate	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
either	NN	O	O
gene	NN	O	O
induction	NN	O	O
or	NN	O	O
gene	NN	O	O
repression	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	O
receptors	NN	O	O
are	NN	O	O
also	NN	O	O
able	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
transcriptional	NN	O	O
factors	NN	O	O
such	NN	O	O
as	NN	O	O
AP-1	NN	O	B-protein
(	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
)	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Through	NN	O	O
these	NN	O	O
actions	NN	O	O
glucocorticosteroids	NN	O	O
are	NN	O	O
susceptible	NN	O	O
to	NN	O	O
modify	NN	O	O
functions	NN	O	O
of	NN	O	O
cells	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
allergic	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
.	NN	O	O

They	NN	O	O
are	NN	O	O
in	NN	O	O
particular	NN	O	O
able	NN	O	O
to	NN	O	O
inhibit	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
pro-inflammatory	NN	O	O
functions	NN	O	O
of	NN	O	O
the	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Jeg-3	NN	O	O
human	NN	O	O
choriocarcinoma-induced	NN	O	O
immunosuppression	NN	O	O
:	NN	O	O
downregulation	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
,	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
alpha-chain	NN	O	I-protein
,	NN	O	O
and	NN	O	O
its	NN	O	O
Jak/Stat	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

PROBLEM	NN	O	O
:	NN	O	O
The	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
and	NN	O	O
immunosuppression-inducing	NN	O	O
capacities	NN	O	O
of	NN	O	O
Jeg-3	NN	O	B-cell_line
human	NN	O	I-cell_line
choriocarcinoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
supernatants	NN	O	O
(	NN	O	O
HCSs	NN	O	O
)	NN	O	O
are	NN	O	O
not	NN	O	O
yet	NN	O	O
completely	NN	O	O
understood	NN	O	O
.	NN	O	O

The	NN	O	O
influence	NN	O	O
on	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
,	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
interferon	NN	O	B-protein
(	NN	O	I-protein
IFN	NN	O	I-protein
)	NN	O	I-protein
-gamma	NN	O	I-protein
production	NN	O	O
;	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
IL-2R	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
-	NN	O	O
,	NN	O	O
beta	NN	O	B-protein
-	NN	O	O
,	NN	O	O
and	NN	O	O
gamma-chain	NN	O	B-protein
;	NN	O	O
and	NN	O	O
the	NN	O	O
signaling	NN	O	B-protein
pathway	NN	O	I-protein
molecules	NN	O	I-protein
Janus	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	B-protein
Jak	NN	O	I-protein
)	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
Jak3	NN	O	B-protein
,	NN	O	O
signal	NN	O	O
transducers	NN	O	O
and	NN	O	O
activators	NN	O	B-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	B-protein
Stat	NN	O	I-protein
)	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Stat5	NN	O	B-protein
should	NN	O	O
be	NN	O	O
investigated	NN	O	O
.	NN	O	O

METHOD	NN	O	O
OF	NN	O	O
STUDY	NN	O	O
:	NN	O	O
For	NN	O	O
assessment	NN	O	O
of	NN	O	O
IL	NN	O	O
production	NN	O	O
,	NN	O	O
whole	NN	O	O
peripheral	NN	O	O
venous	NN	O	O
blood	NN	O	O
from	NN	O	O
healthy	NN	O	O
donors	NN	O	O
was	NN	O	O
stimulated	NN	O	O
with	NN	O	O
phorbol-myristate-acetate	NN	O	O
and	NN	O	O
ionomycine	NN	O	O
.	NN	O	O

Secretion	NN	O	O
of	NN	O	O
ILs	NN	O	B-protein
was	NN	O	O
blocked	NN	O	O
with	NN	O	O
monensine	NN	O	O
.	NN	O	O

Intracellular	NN	O	O
ILs	NN	O	B-protein
were	NN	O	O
analyzed	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

For	NN	O	O
IL-2R	NN	O	B-protein
and	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
molecule	NN	O	O
analysis	NN	O	O
,	NN	O	O
peripheral	NN	O	O
blood	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
phytohemagglutinin	NN	O	B-protein
(	NN	O	O
PHA	NN	O	B-protein
)	NN	O	O
.	NN	O	O

IL-2R	NN	O	O
chains	NN	O	O
were	NN	O	O
measured	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
,	NN	O	O
and	NN	O	O
Jaks/Stats	NN	O	B-protein
by	NN	O	O
sodium	NN	O	O
dodecyl	NN	O	O
sulfate	NN	O	O
polyacrylamide	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
(	NN	O	O
SDS-PAGE	NN	O	O
)	NN	O	O
and	NN	O	O
Western	NN	O	O
blot	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Phorbol-myristate-acetate	NN	O	O
and	NN	O	O
ionomycine	NN	O	O
strongly	NN	O	O
increase	NN	O	O
the	NN	O	O
percent-age	NN	O	O
of	NN	O	O
IL-2+	NN	O	B-cell_line
cells	NN	O	I-cell_line
;	NN	O	O
an	NN	O	O
additional	NN	O	O
50	NN	O	O
%	NN	O	O
HCSs	NN	O	O
significantly	NN	O	O
suppresses	NN	O	O
the	NN	O	O
percentage	NN	O	O
to	NN	O	O
,	NN	O	O
or	NN	O	O
below	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
unstimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

IFN-gamma	NN	O	O
production	NN	O	O
is	NN	O	O
strongly	NN	O	O
decreased	NN	O	O
by	NN	O	O
HCSs	NN	O	O
in	NN	O	O
some	NN	O	O
cases	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
others	NN	O	O
.	NN	O	O

PHA	NN	O	B-protein
stimulates	NN	O	O
IL-2R	NN	O	O
alpha-	NN	O	O
,	NN	O	O
beta-	NN	O	O
,	NN	O	O
and	NN	O	O
gamma-chain	NN	O	O
expression	NN	O	O
and	NN	O	O
their	NN	O	O
signaling	NN	O	B-protein
pathway	NN	O	I-protein
molecules	NN	O	I-protein
Jak1	NN	O	B-protein
,	NN	O	O
Jak3	NN	O	B-protein
,	NN	O	O
Stat1	NN	O	O
,	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Stat5	NN	O	B-protein
.	NN	O	O

50	NN	O	O
%	NN	O	O
HCS	NN	O	O
downregulates	NN	O	O
the	NN	O	O
alpha-chain	NN	O	B-protein
and	NN	O	O
slightly	NN	O	O
upregulates	NN	O	O
the	NN	O	O
beta-chain	NN	O	B-protein
.	NN	O	O

Jak1	NN	O	O
,	NN	O	O
Jak3	NN	O	O
,	NN	O	O
Stat1	NN	O	O
,	NN	O	O
Stat3	NN	O	O
,	NN	O	O
and	NN	O	O
Stat5	NN	O	O
expression	NN	O	O
is	NN	O	O
suppressed	NN	O	O
approximately	NN	O	O
to	NN	O	O
,	NN	O	O
or	NN	O	O
below	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
unstimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
HCS	NN	O	O
forcefully	NN	O	O
blocks	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
;	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-protein
alpha-chain	NN	O	I-protein
;	NN	O	O
and	NN	O	O
Jak1	NN	O	O
,	NN	O	O
Jak3	NN	O	O
,	NN	O	O
Stat1	NN	O	O
,	NN	O	O
Stat3	NN	O	O
,	NN	O	O
and	NN	O	O
Stat5	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
observed	NN	O	O
phenomena	NN	O	O
might	NN	O	O
be	NN	O	O
caused	NN	O	O
by	NN	O	O
downregulation	NN	O	O
of	NN	O	O
an	NN	O	O
IL-2R	NN	O	B-DNA
regulation	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
might	NN	O	O
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
expansion	NN	O	O
of	NN	O	O
choriocarcinoma	NN	O	O
,	NN	O	O
and	NN	O	O
possibly	NN	O	O
in	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
the	NN	O	O
fetal	NN	O	O
allograft	NN	O	O
.	NN	O	O

-DOCSTART-	O

Apoptosis-resistant	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
have	NN	O	O
a	NN	O	O
deficiency	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
transcription	NN	O	O
.	NN	O	O

Apoptosis	NN	O	O
induced	NN	O	O
through	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
in	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
mostly	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
FasL	NN	O	B-protein
)	NN	O	O
as	NN	O	O
a	NN	O	O
primary	NN	O	O
event	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
commitment	NN	O	O
to	NN	O	O
death	NN	O	O
.	NN	O	O

To	NN	O	O
gain	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
events	NN	O	O
that	NN	O	O
regulate	NN	O	O
this	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
took	NN	O	O
advantage	NN	O	O
of	NN	O	O
our	NN	O	O
previously	NN	O	O
described	NN	O	O
mutant	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
cells	NN	O	O
are	NN	O	O
deficient	NN	O	O
in	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
upon	NN	O	O
TCR	NN	O	B-protein
triggering	NN	O	O
,	NN	O	O
although	NN	O	O
their	NN	O	O
cytokine	NN	O	O
(	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
production	NN	O	O
is	NN	O	O
normal	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
both	NN	O	O
a	NN	O	O
FasL-	NN	O	B-DNA
and	NN	O	I-DNA
a	NN	O	I-DNA
consensus	NN	O	I-DNA
NF-kappaB-	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
are	NN	O	O
inefficiently	NN	O	O
induced	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
compared	NN	O	O
to	NN	O	O
wild-type	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
inducible	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
FasL	NN	O	B-DNA
reporter	NN	O	I-DNA
is	NN	O	O
abolished	NN	O	O
by	NN	O	O
specific	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
we	NN	O	O
could	NN	O	O
trace	NN	O	O
the	NN	O	O
deficit	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
an	NN	O	O
inefficient	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
evidencing	NN	O	O
a	NN	O	O
relevant	NN	O	O
role	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
versus	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
is	NN	O	O
differentially	NN	O	O
sensitive	NN	O	O
to	NN	O	O
NF-kappaB	NN	O	B-protein
deprivation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Decreased	NN	O	O
proteasome	NN	O	B-protein
-mediated	NN	O	O
degradation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
elderly	NN	O	O
:	NN	O	O
A	NN	O	O
role	NN	O	O
in	NN	O	O
immune	NN	O	O
senescence	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
is	NN	O	O
a	NN	O	O
highly	NN	O	O
regulated	NN	O	O
process	NN	O	O
requiring	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
ubiquitination	NN	O	O
,	NN	O	O
and	NN	O	O
proteasome	NN	O	B-protein
-mediated	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
cytosolic	NN	O	B-protein
inhibitor	NN	O	I-protein
IkappaBalpha	NN	O	B-protein
.	NN	O	O

Analyses	NN	O	O
of	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
in	NN	O	O
TNF-alpha-treated	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
young	NN	O	O
and	NN	O	O
elderly	NN	O	O
donors	NN	O	O
revealed	NN	O	O
severely	NN	O	O
compromised	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
elderly	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
activation-induced	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
ubiquitination	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
did	NN	O	O
not	NN	O	O
demonstrate	NN	O	O
any	NN	O	O
significant	NN	O	O
age-related	NN	O	O
alterations	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
examination	NN	O	O
of	NN	O	O
proteasome	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
these	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
fluorogenic	NN	O	O
peptide	NN	O	O
assays	NN	O	O
revealed	NN	O	O
a	NN	O	O
significant	NN	O	O
age-related	NN	O	O
decline	NN	O	O
in	NN	O	O
chymotryptic	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
decline	NN	O	O
in	NN	O	O
proteasome	NN	O	B-protein
activity	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
failure	NN	O	O
to	NN	O	O
fully	NN	O	O
degrade	NN	O	O
IkappaBalpha	NN	O	B-protein
in	NN	O	O
the	NN	O	O
elderly	NN	O	O
.	NN	O	O

This	NN	O	O
failure	NN	O	O
to	NN	O	O
degrade	NN	O	O
IkappaBalpha	NN	O	B-protein
may	NN	O	O
underlie	NN	O	O
both	NN	O	O
the	NN	O	O
observed	NN	O	O
decrease	NN	O	O
in	NN	O	O
NFkappaB	NN	O	B-protein
induction	NN	O	O
and	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	O
expression	NN	O	O
in	NN	O	O
TNF-treated	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
during	NN	O	O
aging	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
decreased	NN	O	O
proteasome	NN	O	B-protein
-mediated	NN	O	O
degradation	NN	O	O
may	NN	O	O
be	NN	O	O
central	NN	O	O
to	NN	O	O
immune	NN	O	O
dysfunction	NN	O	O
that	NN	O	O
accompanies	NN	O	O
aging	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
-inducible	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
by	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
interferons	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Th2-type	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interleukin-13	NN	O	B-protein
(	NN	O	O
IL-13	NN	O	B-protein
)	NN	O	O
,	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
distinct	NN	O	O
subset	NN	O	O
of	NN	O	O
genes	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
including	NN	O	O
FcepsilonRIIb	NN	O	B-protein
(	NN	O	O
CD23	NN	O	B-protein
)	NN	O	O
,	NN	O	O
15-lipoxygenase	NN	O	B-protein
,	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	I-protein
antagonist	NN	O	I-protein
(	NN	O	O
IL-1ra	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
IL-1	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
IL-1R	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Type	NN	O	B-protein
I	NN	O	I-protein
interferons	NN	O	I-protein
(	NN	O	O
IFN-alpha	NN	O	B-protein
and	NN	O	O
IFN-beta	NN	O	B-protein
)	NN	O	O
and	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
interferon	NN	O	I-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
inhibit	NN	O	O
induction	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
IFNs	NN	O	B-protein
mediate	NN	O	O
this	NN	O	O
inhibition	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
defined	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
overview	NN	O	O
,	NN	O	O
we	NN	O	O
discuss	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
STAT6	NN	O	B-protein
(	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription-6	NN	O	I-protein
)	NN	O	O
in	NN	O	O
mediating	NN	O	O
IL-4-	NN	O	O
and	NN	O	O
IL-13-induced	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
also	NN	O	O
discuss	NN	O	O
our	NN	O	O
recent	NN	O	O
findings	NN	O	O
that	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
IFNs	NN	O	I-protein
suppress	NN	O	O
IL-4	NN	O	B-protein
/	NN	O	O
IL-13	NN	O	B-protein
-inducible	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
IFNs	NN	O	I-protein
to	NN	O	O
inhibit	NN	O	O
IL-4	NN	O	B-protein
/	NN	O	O
IL-13	NN	O	B-protein
-induced	NN	O	O
STAT6	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
dose-	NN	O	O
and	NN	O	O
time-dependent	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
unique	NN	O	O
to	NN	O	O
monocytes	NN	O	B-cell_type
because	NN	O	O
IFNs	NN	O	B-protein
induce	NN	O	O
the	NN	O	O
same	NN	O	O
effects	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
not	NN	O	O
evident	NN	O	O
unless	NN	O	O
cells	NN	O	O
are	NN	O	O
preincubated	NN	O	O
with	NN	O	O
IFN	NN	O	B-protein
for	NN	O	O
at	NN	O	O
least	NN	O	O
1	NN	O	O
h	NN	O	O
before	NN	O	O
IL-4	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
inhibition	NN	O	O
can	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
requirement	NN	O	O
for	NN	O	O
de	NN	O	O
novo	NN	O	O
transcription	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
stimulation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
by	NN	O	O
IFN	NN	O	B-protein
activates	NN	O	O
de	NN	O	O
novo	NN	O	O
synthesis	NN	O	O
of	NN	O	O
an	NN	O	O
inhibitory	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
possibly	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
SOCS/	NN	O	B-protein
SSI/CIS	NN	O	I-protein
gene	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
capable	NN	O	O
of	NN	O	O
suppressing	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
.	NN	O	O

Because	NN	O	O
STAT6	NN	O	B-protein
activation	NN	O	O
plays	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
in	NN	O	O
IL-4	NN	O	B-protein
/	NN	O	O
IL-13	NN	O	B-protein
-induced	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
IFN-beta	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
to	NN	O	O
inhibit	NN	O	O
STAT6	NN	O	B-protein
activity	NN	O	O
provides	NN	O	O
an	NN	O	O
explanation	NN	O	O
for	NN	O	O
how	NN	O	O
IFNs	NN	O	B-protein
can	NN	O	O
suppress	NN	O	O
IL-4	NN	O	B-protein
/	NN	O	O
IL-13	NN	O	B-protein
-inducible	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

HLA	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
involves	NN	O	O
cyclin	NN	O	B-DNA
E	NN	O	I-DNA
-mediated	NN	O	O
inactivation	NN	O	O
of	NN	O	O
Rb	NN	O	O
function	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Chronic	NN	O	O
rejection	NN	O	O
of	NN	O	O
transplanted	NN	O	O
organs	NN	O	O
is	NN	O	O
manifested	NN	O	O
as	NN	O	O
atherosclerosis	NN	O	O
of	NN	O	O
the	NN	O	O
blood	NN	O	O
vessels	NN	O	O
of	NN	O	O
the	NN	O	O
allograft	NN	O	O
.	NN	O	O

HLA	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
Ags	NN	O	I-protein
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
this	NN	O	O
process	NN	O	O
,	NN	O	O
since	NN	O	O
signaling	NN	O	O
via	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
molecules	NN	O	I-protein
can	NN	O	O
induce	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
aortic	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
smooth	NN	O	B-cell_type
muscle	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
correlates	NN	O	O
with	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
Rb	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
as	NN	O	O
human	NN	O	B-cell_type
aortic	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

HLA	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
-mediated	NN	O	O
inactivation	NN	O	O
of	NN	O	O
Rb	NN	O	B-protein
can	NN	O	O
be	NN	O	O
inhibited	NN	O	O
specifically	NN	O	O
by	NN	O	O
neutralizing	NN	O	B-protein
Abs	NN	O	I-protein
to	NN	O	O
basic	NN	O	B-protein
fibroblast	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
bFGF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
FGF	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
process	NN	O	O
.	NN	O	O

Signaling	NN	O	O
through	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
molecules	NN	O	I-protein
induced	NN	O	O
cyclin	NN	O	B-DNA
E	NN	O	I-DNA
-associated	NN	O	O
kinase	NN	O	O
activity	NN	O	O
within	NN	O	O
4	NN	O	O
h	NN	O	O
in	NN	O	O
quiescent	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
appeared	NN	O	O
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
inactivation	NN	O	O
of	NN	O	O
Rb	NN	O	B-protein
.	NN	O	O

A	NN	O	O
cdk2	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
Olomoucine	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
dominant-negative	NN	O	B-DNA
cdk2	NN	O	I-DNA
construct	NN	O	I-DNA
prevented	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
-mediated	NN	O	O
inactivation	NN	O	O
of	NN	O	O
Rb	NN	O	B-protein
;	NN	O	O
in	NN	O	O
contrast	NN	O	O
,	NN	O	O
dominant-negative	NN	O	B-DNA
cdk4	NN	O	I-DNA
and	NN	O	I-DNA
cdk6	NN	O	I-DNA
constructs	NN	O	I-DNA
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
increase	NN	O	O
in	NN	O	O
cyclin	NN	O	B-protein
D-associated	NN	O	I-protein
kinase	NN	O	I-protein
activity	NN	O	O
upon	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
ligation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
cyclin	NN	O	B-protein
E-dependent	NN	O	I-protein
kinase	NN	O	I-protein
activity	NN	O	O
mediates	NN	O	O
Rb	NN	O	B-protein
inactivation	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
E2F	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Immunosuppressant	NN	O	O
PG490	NN	O	O
(	NN	O	O
triptolide	NN	O	O
)	NN	O	O
inhibits	NN	O	O
T-cell	NN	O	B-protein
interleukin-2	NN	O	I-protein
expression	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
purine-box/nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	O
activated	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
and	NN	O	O
NF-kappaB	NN	O	B-protein
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

PG490	NN	O	O
(	NN	O	O
triptolide	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
diterpene	NN	O	O
triepoxide	NN	O	O
with	NN	O	O
potent	NN	O	O
immunosuppressive	NN	O	O
and	NN	O	O
antiinflammatory	NN	O	O
properties	NN	O	O
.	NN	O	O

PG490	NN	O	O
inhibits	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
antibody	NN	O	O
to	NN	O	O
CD3	NN	O	B-protein
(	NN	O	O
IC50	NN	O	O
of	NN	O	O
10	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
with	NN	O	O
PMA	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
(	NN	O	O
Iono	NN	O	O
,	NN	O	O
IC50	NN	O	O
of	NN	O	O
40	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
Jurkat	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
,	NN	O	O
PG490	NN	O	O
inhibits	NN	O	O
PMA/Iono-stimulated	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

PG490	NN	O	O
inhibits	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
at	NN	O	O
the	NN	O	O
purine-box/antigen	NN	O	B-DNA
receptor	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
ARRE	NN	O	I-DNA
)	NN	O	I-DNA
/nuclear	NN	O	I-DNA
factor	NN	O	I-DNA
of	NN	O	I-DNA
activated	NN	O	I-DNA
T-cells	NN	O	I-DNA
(	NN	O	I-DNA
NF-AT	NN	O	I-DNA
)	NN	O	I-DNA
target	NN	O	I-DNA
sequence	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
at	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

PG490	NN	O	O
can	NN	O	O
completely	NN	O	O
inhibit	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
at	NN	O	O
the	NN	O	O
purine-box/ARRE/NF-AT	NN	O	B-DNA
and	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
target	NN	O	I-DNA
DNA	NN	O	I-DNA
sequences	NN	O	I-DNA
triggered	NN	O	O
by	NN	O	O
all	NN	O	O
stimuli	NN	O	O
examined	NN	O	O
(	NN	O	O
PMA	NN	O	O
,	NN	O	O
PMA/Iono	NN	O	O
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
)	NN	O	O
.	NN	O	O

PG490	NN	O	O
also	NN	O	O
inhibits	NN	O	O
PMA-stimulated	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
chimeric	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
which	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
TA1	NN	O	I-protein
transactivation	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
NF-kappaB	NN	O	B-protein
p65	NN	O	I-protein
is	NN	O	O
fused	NN	O	O
to	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
GAL4	NN	O	B-protein
.	NN	O	O

In	NN	O	O
16HBE	NN	O	B-cell_type
human	NN	O	I-cell_type
bronchial	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
IL-8	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
regulated	NN	O	O
predominantly	NN	O	O
by	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
PG490	NN	O	O
but	NN	O	O
not	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
can	NN	O	O
completely	NN	O	O
inhibit	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
PG490	NN	O	O
inhibition	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
differs	NN	O	O
from	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
involves	NN	O	O
nuclear	NN	O	O
inhibition	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
the	NN	O	O
purine-box	NN	O	B-DNA
regulator	NN	O	I-DNA
operating	NN	O	O
at	NN	O	O
the	NN	O	O
ARRE/NF-AT	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
a	NN	O	O
step	NN	O	O
after	NN	O	O
specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

-DOCSTART-	O

Unexpected	NN	O	O
and	NN	O	O
coordinated	NN	O	O
expression	NN	O	O
of	NN	O	O
Spi-1	NN	O	B-protein
,	NN	O	O
Fli-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
four	NN	O	O
Epo-dependent	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
established	NN	O	O
from	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
displaying	NN	O	O
erythroid-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
thermosensitive	NN	O	B-protein
SV40	NN	O	I-protein
T	NN	O	I-protein
antigen	NN	O	I-protein
.	NN	O	O

Most	NN	O	O
erythroleukemic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
established	NN	O	O
in	NN	O	O
vitro	NN	O	O
coexpress	NN	O	O
erythrocytic	NN	O	B-protein
and	NN	O	I-protein
megakaryocytic	NN	O	I-protein
markers	NN	O	I-protein
that	NN	O	O
often	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
expression	NN	O	O
of	NN	O	O
Spi-1	NN	O	B-protein
and/or	NN	O	O
Fli-1	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
known	NN	O	O
as	NN	O	O
transactivators	NN	O	B-protein
of	NN	O	I-protein
megakaryocyte-specific	NN	O	I-protein
promoters	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
establishing	NN	O	O
new	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
keeping	NN	O	O
strictly	NN	O	O
erythroid-specific	NN	O	O
properties	NN	O	O
in	NN	O	O
vitro	NN	O	O
through	NN	O	O
the	NN	O	O
targeted	NN	O	O
and	NN	O	O
conditional	NN	O	O
immortalization	NN	O	O
of	NN	O	O
erythrocytic	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

For	NN	O	O
that	NN	O	O
purpose	NN	O	O
,	NN	O	O
we	NN	O	O
established	NN	O	O
several	NN	O	O
lines	NN	O	O
of	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
displaying	NN	O	O
erythroid-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
thermosensitive	NN	O	B-protein
SV40	NN	O	I-protein
T	NN	O	I-protein
antigen	NN	O	I-protein
.	NN	O	O

As	NN	O	O
expected	NN	O	O
,	NN	O	O
these	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
developed	NN	O	O
splenomegaly	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
massive	NN	O	O
amplification	NN	O	O
of	NN	O	O
Ter	NN	O	B-cell_line
119	NN	O	I-cell_line
positive	NN	O	I-cell_line
erythroid	NN	O	I-cell_line
nucleated	NN	O	I-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
T	NN	O	B-protein
antigen	NN	O	I-protein
.	NN	O	O

Despite	NN	O	O
this	NN	O	O
drastic	NN	O	O
effect	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
immortalization	NN	O	O
of	NN	O	O
erythropoietin-dependent	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
unexpectedly	NN	O	O
occurred	NN	O	O
at	NN	O	O
low	NN	O	O
frequency	NN	O	O
,	NN	O	O
and	NN	O	O
all	NN	O	O
four	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
established	NN	O	O
expressed	NN	O	O
both	NN	O	O
erythrocytic	NN	O	B-protein
(	NN	O	I-protein
globins	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
megakaryocytic	NN	O	I-protein
markers	NN	O	I-protein
(	NN	O	O
glycoprotein	NN	O	B-protein
IIb	NN	O	I-protein
,	NN	O	O
platelet	NN	O	B-protein
factor	NN	O	I-protein
4	NN	O	I-protein
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
Spi-1	NN	O	B-protein
and	NN	O	O
Fli-1	NN	O	B-protein
transcripts	NN	O	I-protein
at	NN	O	O
permissive	NN	O	O
temperature	NN	O	O
.	NN	O	O

Switching	NN	O	O
the	NN	O	O
cells	NN	O	O
to	NN	O	O
the	NN	O	O
nonpermissive	NN	O	O
temperature	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
marked	NN	O	O
increase	NN	O	O
in	NN	O	O
globin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
and	NN	O	O
concomitant	NN	O	O
decrease	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
Spi-1	NN	O	B-protein
,	NN	O	O
Fli-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
an	NN	O	O
erythropoietin	NN	O	B-protein
-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
Spi-1	NN	O	B-DNA
and	NN	O	I-DNA
Fli-1	NN	O	I-DNA
genes	NN	O	I-DNA
already	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
Ter	NN	O	B-cell_line
119	NN	O	I-cell_line
positive	NN	O	I-cell_line
cell	NN	O	I-cell_line
population	NN	O	I-cell_line
of	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
spleen	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
like	NN	O	O
normal	NN	O	B-cell_line
Ter	NN	O	I-cell_line
119	NN	O	I-cell_line
erythroid	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
these	NN	O	O
Ter	NN	O	B-cell_line
119	NN	O	I-cell_line
positive	NN	O	I-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
still	NN	O	O
expressed	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
beta-globin	NN	O	B-protein
and	NN	O	O
very	NN	O	O
low	NN	O	O
or	NN	O	O
undetectable	NN	O	O
glycoprotein	NN	O	B-protein
IIb	NN	O	I-protein
and	NN	O	O
platelet	NN	O	B-protein
factor	NN	O	I-protein
4	NN	O	I-protein
megakaryocytic	NN	O	I-protein
transcripts	NN	O	I-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
unexpected	NN	O	O
expression	NN	O	O
of	NN	O	O
megakaryocytic	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
specific	NN	O	O
property	NN	O	O
of	NN	O	O
immortalized	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
only	NN	O	O
by	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
Spi-1	NN	O	B-DNA
and/or	NN	O	I-DNA
Fli-1	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Fludarabine-induced	NN	O	O
immunosuppression	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
inhibition	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Fludarabine	NN	O	O
is	NN	O	O
a	NN	O	O
nucleoside	NN	O	O
analog	NN	O	O
used	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
hematologic	NN	O	O
malignancies	NN	O	O
that	NN	O	O
can	NN	O	O
induce	NN	O	O
severe	NN	O	O
and	NN	O	O
prolonged	NN	O	O
immunosuppression	NN	O	O
.	NN	O	O

Although	NN	O	O
it	NN	O	O
can	NN	O	O
be	NN	O	O
incorporated	NN	O	O
into	NN	O	O
the	NN	O	O
DNA	NN	O	O
of	NN	O	O
dividing	NN	O	O
cells	NN	O	O
,	NN	O	O
fludarabine	NN	O	O
is	NN	O	O
also	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
a	NN	O	O
low	NN	O	O
growth	NN	O	O
fraction	NN	O	O
,	NN	O	O
thus	NN	O	O
it	NN	O	O
must	NN	O	O
have	NN	O	O
other	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
action	NN	O	O
.	NN	O	O

STAT1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
many	NN	O	O
lymphocyte-activating	NN	O	B-protein
cytokines	NN	O	I-protein
including	NN	O	O
the	NN	O	O
interferons	NN	O	B-protein
,	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
cell-mediated	NN	O	O
immunity	NN	O	O
,	NN	O	O
as	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
prominent	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
control	NN	O	O
viral	NN	O	O
infections	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
fludarabine	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
immunosuppressant	NN	O	O
cyclosporine	NN	O	O
A	NN	O	O
,	NN	O	O
inhibits	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT1	NN	O	B-protein
-dependent	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
resting	NN	O	I-cell_type
or	NN	O	I-cell_type
activated	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Fludarabine	NN	O	O
caused	NN	O	O
a	NN	O	O
specific	NN	O	O
depletion	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
and	NN	O	O
mRNA	NN	O	O
)	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
other	NN	O	O
STATs	NN	O	B-protein
.	NN	O	O

This	NN	O	O
loss	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
was	NN	O	O
also	NN	O	O
seen	NN	O	O
in	NN	O	O
cells	NN	O	O
from	NN	O	O
patients	NN	O	O
treated	NN	O	O
with	NN	O	O
fludarabine	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Brief	NN	O	O
exposure	NN	O	O
to	NN	O	O
fludarabine	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
sustained	NN	O	O
loss	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	O
analogous	NN	O	O
to	NN	O	O
the	NN	O	O
prolonged	NN	O	O
period	NN	O	O
of	NN	O	O
immunosuppression	NN	O	O
induced	NN	O	O
by	NN	O	O
exposure	NN	O	O
to	NN	O	O
the	NN	O	O
drug	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
STAT1	NN	O	B-protein
may	NN	O	O
be	NN	O	O
a	NN	O	O
useful	NN	O	O
target	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
new	NN	O	O
immunosuppressive	NN	O	O
and	NN	O	O
antineoplastic	NN	O	O
agents	NN	O	O
.	NN	O	O

-DOCSTART-	O

Studies	NN	O	O
into	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
phosphatase	NN	O	I-protein
inhibitor	NN	O	O
phenylarsine	NN	O	O
oxide	NN	O	O
on	NN	O	O
NFkappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
during	NN	O	O
aging	NN	O	O
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
altered	NN	O	O
IkappaB-alpha	NN	O	B-protein
phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
Factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NFkappaB	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
regulator	NN	O	O
of	NN	O	O
several	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
stimuli	NN	O	O
,	NN	O	O
including	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
active	NN	O	O
p65-50	NN	O	B-protein
heterodimer	NN	O	I-protein
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
albeit	NN	O	O
at	NN	O	O
a	NN	O	O
lower	NN	O	O
level	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
elderly	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
that	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
PAO	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
TNF-alpha	NN	O	B-protein
treated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
PAO-sensitive	NN	O	B-protein
phosphatase	NN	O	I-protein
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NFkappaB	NN	O	B-protein
via	NN	O	O
this	NN	O	O
pathway	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
it	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
aging	NN	O	O
does	NN	O	O
not	NN	O	O
influence	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
this	NN	O	O
phosphatase	NN	O	B-protein
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
DMP	NN	O	O
prior	NN	O	O
to	NN	O	O
treatment	NN	O	O
with	NN	O	O
PAO	NN	O	O
and	NN	O	O
TNF	NN	O	B-protein
abolishes	NN	O	O
the	NN	O	O
inhibition	NN	O	O
induced	NN	O	O
by	NN	O	O
PAO	NN	O	O
,	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
both	NN	O	O
young	NN	O	O
and	NN	O	O
old	NN	O	O
donors	NN	O	O
,	NN	O	O
alike	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
failure	NN	O	O
to	NN	O	O
degrade	NN	O	O
IkappaB-alpha	NN	O	B-protein
in	NN	O	O
cytosols	NN	O	O
of	NN	O	O
TNF-treated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
pretreated	NN	O	O
with	NN	O	O
PAO	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
its	NN	O	O
interference	NN	O	O
with	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
and	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
its	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
proteasomal	NN	O	O
degradation	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
collectively	NN	O	O
suggest	NN	O	O
that	NN	O	O
PAO	NN	O	O
interferes	NN	O	O
with	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
the	NN	O	O
regulated	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
,	NN	O	O
induced	NN	O	O
by	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
without	NN	O	O
affecting	NN	O	O
the	NN	O	O
chymotryptic	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
,	NN	O	O
independent	NN	O	O
of	NN	O	O
age	NN	O	O
.	NN	O	O

-DOCSTART-	O

Spi-C	NN	O	B-protein
,	NN	O	O
a	NN	O	O
novel	NN	O	O
Ets	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
is	NN	O	O
temporally	NN	O	O
regulated	NN	O	O
during	NN	O	O
B	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
.	NN	O	O

A	NN	O	O
novel	NN	O	O
Ets	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
isolated	NN	O	O
by	NN	O	O
yeast	NN	O	O
one-hybrid	NN	O	O
screening	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
library	NN	O	I-DNA
made	NN	O	O
from	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-cell_line
mouse	NN	O	I-cell_line
splenic	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
using	NN	O	O
the	NN	O	O
SP6	NN	O	B-DNA
kappa	NN	O	I-DNA
promoter	NN	O	I-DNA
kappaY	NN	O	I-DNA
element	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
bait	NN	O	O
.	NN	O	O

The	NN	O	O
novel	NN	O	O
Ets	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
most	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
PU.1	NN	O	B-protein
and	NN	O	O
Spi-B	NN	O	B-protein
within	NN	O	O
the	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
Ets	NN	O	I-DNA
domain	NN	O	I-DNA
and	NN	O	O
was	NN	O	O
therefore	NN	O	O
named	NN	O	O
Spi-C	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
Spi-C	NN	O	B-protein
may	NN	O	O
represent	NN	O	O
a	NN	O	O
novel	NN	O	O
subgroup	NN	O	O
within	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
protein	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
as	NN	O	O
it	NN	O	O
differed	NN	O	O
significantly	NN	O	O
from	NN	O	O
Spi-B	NN	O	B-protein
and	NN	O	O
PU.1	NN	O	B-protein
within	NN	O	O
helix	NN	O	B-protein
1	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

Spi-C	NN	O	B-protein
was	NN	O	O
encoded	NN	O	O
by	NN	O	O
a	NN	O	O
single-copy	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
was	NN	O	O
mapped	NN	O	O
to	NN	O	O
chromosome	NN	O	B-DNA
10	NN	O	I-DNA
,	NN	O	O
region	NN	O	B-DNA
C	NN	O	I-DNA
.	NN	O	O

Spi-C	NN	O	B-protein
interacted	NN	O	O
with	NN	O	O
DNA	NN	O	O
similarly	NN	O	O
to	NN	O	O
PU.1	NN	O	B-protein
as	NN	O	O
judged	NN	O	O
by	NN	O	O
methylation	NN	O	O
interference	NN	O	O
,	NN	O	O
band-shift	NN	O	O
and	NN	O	O
site	NN	O	O
selection	NN	O	O
analysis	NN	O	O
,	NN	O	O
and	NN	O	O
activated	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
kappaY	NN	O	B-DNA
element	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
upon	NN	O	O
co-transfection	NN	O	O
of	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Spi-C	NN	O	B-RNA
RNA	NN	O	I-RNA
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
at	NN	O	O
lower	NN	O	O
levels	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
pre-B	NN	O	B-cell_line
cell	NN	O	I-cell_line
and	NN	O	I-cell_line
plasma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
Spi-C	NN	O	B-protein
-negative	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
Spi-C	NN	O	B-protein
might	NN	O	O
be	NN	O	O
a	NN	O	O
regulatory	NN	O	B-protein
molecule	NN	O	I-protein
during	NN	O	O
a	NN	O	O
specific	NN	O	O
phase	NN	O	O
of	NN	O	O
B	NN	O	O
lymphoid	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
requires	NN	O	O
autoinduction	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
negative	NN	O	O
autoregulation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
seen	NN	O	O
in	NN	O	O
most	NN	O	O
cells	NN	O	O
and	NN	O	O
tissues	NN	O	O
,	NN	O	O
GR	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
positively	NN	O	O
autoregulated	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
other	NN	O	O
cells	NN	O	O
sensitive	NN	O	O
to	NN	O	O
glucocorticoid-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
positive	NN	O	O
autoregulation	NN	O	O
is	NN	O	O
a	NN	O	O
necessary	NN	O	O
component	NN	O	O
of	NN	O	O
glucocorticoid-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
,	NN	O	O
a	NN	O	O
wild-type	NN	O	B-DNA
GR	NN	O	I-DNA
gene	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
a	NN	O	O
tetracycline-regulated	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
stably	NN	O	O
transfected	NN	O	O
into	NN	O	O
glucocorticoid-resistant	NN	O	B-cell_line
cells	NN	O	I-cell_line
lacking	NN	O	O
endogenous	NN	O	O
functional	NN	O	O
receptor	NN	O	O
.	NN	O	O

Transfectants	NN	O	B-cell_line
grown	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
tetracycline	NN	O	O
contained	NN	O	O
about	NN	O	O
15	NN	O	O
,	NN	O	O
000	NN	O	O
receptors/cell	NN	O	O
,	NN	O	O
a	NN	O	O
value	NN	O	O
approximately	NN	O	O
equal	NN	O	O
to	NN	O	O
basal	NN	O	O
level	NN	O	O
GR	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
glucocorticoid-sensitive	NN	O	B-cell_line
6TG1.1	NN	O	I-cell_line
cells	NN	O	I-cell_line
before	NN	O	O
steroid	NN	O	O
treatment	NN	O	O
.	NN	O	O

Under	NN	O	O
these	NN	O	O
conditions	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
had	NN	O	O
a	NN	O	O
minimal	NN	O	O
effect	NN	O	O
on	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
elicited	NN	O	O
little	NN	O	O
internucleosomal	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
,	NN	O	O
and	NN	O	O
induced	NN	O	O
no	NN	O	O
cell	NN	O	O
cycle	NN	O	O
perturbation	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
tetracycline	NN	O	O
,	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
increased	NN	O	O
2-3-fold	NN	O	O
,	NN	O	O
and	NN	O	O
cells	NN	O	O
expressed	NN	O	O
48	NN	O	O
,	NN	O	O
000	NN	O	O
receptors	NN	O	O
,	NN	O	O
a	NN	O	O
level	NN	O	O
nearly	NN	O	O
equivalent	NN	O	O
to	NN	O	O
that	NN	O	O
present	NN	O	O
in	NN	O	O
6TG1.1	NN	O	B-cell_line
cells	NN	O	I-cell_line
after	NN	O	O
18	NN	O	O
h	NN	O	O
of	NN	O	O
autoinduction	NN	O	O
.	NN	O	O

Under	NN	O	O
these	NN	O	O
conditions	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
markedly	NN	O	O
inhibited	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
caused	NN	O	O
G1	NN	O	O
arrest	NN	O	O
,	NN	O	O
and	NN	O	O
induced	NN	O	O
significant	NN	O	O
internucleosomal	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
therefore	NN	O	O
suggest	NN	O	O
that	NN	O	O
basal	NN	O	O
level	NN	O	O
GR	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
inadequate	NN	O	O
to	NN	O	O
mediate	NN	O	O
glucocorticoid-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
glucocorticoid-sensitive	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
that	NN	O	O
positive	NN	O	O
autoregulation	NN	O	O
is	NN	O	O
a	NN	O	O
necessary	NN	O	O
component	NN	O	O
of	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
adult	NN	O	I-cell_type
T-cell	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
etiologic	NN	O	O
agent	NN	O	O
of	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ATL	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
viral	NN	O	B-protein
protein	NN	O	I-protein
Tax	NN	O	B-protein
induces	NN	O	O
the	NN	O	O
activation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocalization	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
proposed	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
transformation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
genes	NN	O	I-DNA
including	NN	O	O
Tax	NN	O	B-protein
are	NN	O	O
not	NN	O	O
expressed	NN	O	O
significantly	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
ATL	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
basis	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
ATL	NN	O	O
patients	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
ATL	NN	O	O
patients	NN	O	O
,	NN	O	O
like	NN	O	O
HTLV-I-infected	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
display	NN	O	O
constitutive	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
increased	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
(	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Whereas	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
Tax-expressing	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
consisted	NN	O	O
mostly	NN	O	O
of	NN	O	O
p50/c-Rel	NN	O	B-protein
,	NN	O	O
fresh	NN	O	O
ATL	NN	O	O
samples	NN	O	O
contained	NN	O	O
p50/p50	NN	O	B-protein
and	NN	O	I-protein
p50/p65	NN	O	I-protein
heterodimers	NN	O	I-protein
.	NN	O	O

One	NN	O	O
T-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
ATL	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
TL-Om1	NN	O	O
,	NN	O	O
displayed	NN	O	O
constitutive	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
enhanced	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
,	NN	O	O
despite	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
detectable	NN	O	O
Tax	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
TL-Om1	NN	O	O
consists	NN	O	O
of	NN	O	O
p50/p50	NN	O	B-protein
and	NN	O	O
p50/p65	NN	O	B-protein
like	NN	O	O
that	NN	O	O
in	NN	O	O
fresh	NN	O	O
primary	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
occurs	NN	O	O
through	NN	O	O
a	NN	O	O
Tax	NN	O	B-protein
-independent	NN	O	O
mechanism	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
ATL	NN	O	O
patients	NN	O	O
,	NN	O	O
possibly	NN	O	O
due	NN	O	O
to	NN	O	O
differential	NN	O	O
NF-kappaB	NN	O	B-protein
subunit	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interferon-alpha	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
STATs1	NN	O	B-protein
,	NN	O	I-protein
-3	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
growth	NN	O	O
arrest	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
Lck	NN	O	B-protein
and	NN	O	O
active	NN	O	O
mitogen-activated	NN	O	B-protein
kinase	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Type	NN	O	O
I	NN	O	O
interferons	NN	O	B-protein
(	NN	O	O
IFNs	NN	O	B-protein
)	NN	O	O
are	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
have	NN	O	O
antiviral	NN	O	O
and	NN	O	O
antiproliferative	NN	O	O
effects	NN	O	O
.	NN	O	O

Data	NN	O	O
regarding	NN	O	O
the	NN	O	O
processes	NN	O	O
by	NN	O	O
which	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
transduce	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
cell	NN	O	O
membrane	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
are	NN	O	O
becoming	NN	O	O
increasingly	NN	O	O
complex	NN	O	O
.	NN	O	O

The	NN	O	O
most	NN	O	O
characterized	NN	O	O
pathway	NN	O	O
is	NN	O	O
via	NN	O	O
JAK	NN	O	B-protein
-STAT	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
established	NN	O	O
a	NN	O	O
potential	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
Src-family	NN	O	B-protein
kinase	NN	O	I-protein
Lck	NN	O	B-protein
in	NN	O	O
JAK	NN	O	B-protein
-STAT	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
designed	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Lck	NN	O	B-protein
in	NN	O	O
IFN-alpha	NN	O	B-protein
signaling	NN	O	O
by	NN	O	O
using	NN	O	O
the	NN	O	O
Jurkat	NN	O	O
,	NN	O	O
JCam	NN	O	O
(	NN	O	O
an	NN	O	O
Lck-defective	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
Jurkat	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
and	NN	O	O
JCam/Lck	NN	O	B-cell_line
(	NN	O	O
JCam	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
Lck	NN	O	B-protein
restored	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
IFN-alpha	NN	O	B-protein
can	NN	O	O
induce	NN	O	O
MAPK	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
in	NN	O	O
cells	NN	O	O
containing	NN	O	O
Lck	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
STATs1	NN	O	B-protein
and	NN	O	I-protein
-3	NN	O	I-protein
are	NN	O	O
effectively	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
activated	NN	O	O
to	NN	O	O
bind	NN	O	O
DNA	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
Lck	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
IFN-alpha-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
IFN-alpha	NN	O	B-protein
exerts	NN	O	O
an	NN	O	O
antiproliferative	NN	O	O
effect	NN	O	O
in	NN	O	O
all	NN	O	O
three	NN	O	O
cell	NN	O	O
lines	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
Lck	NN	O	B-protein
and	NN	O	O
active	NN	O	O
MAPK	NN	O	B-protein
do	NN	O	O
not	NN	O	O
affect	NN	O	O
IFN-alpha	NN	O	B-protein
-induced	NN	O	O
growth	NN	O	O
arrest	NN	O	O
or	NN	O	O
induction	NN	O	O
of	NN	O	O
STAT1s1	NN	O	O
and	NN	O	O
-3	NN	O	O
DNA	NN	O	O
binding	NN	O	O
ability	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Selective	NN	O	O
activation	NN	O	O
and	NN	O	O
functional	NN	O	O
significance	NN	O	O
of	NN	O	O
p38alpha	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
in	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-cell_line
neutrophils	NN	O	I-cell_line
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
by	NN	O	O
proinflammatory	NN	O	O
stimuli	NN	O	O
selectively	NN	O	O
initiates	NN	O	O
intracellular	NN	O	O
signal	NN	O	O
transduction	NN	O	O
via	NN	O	O
sequential	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
kinases	NN	O	B-protein
.	NN	O	O

Lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
is	NN	O	O
known	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPk	NN	O	B-protein
)	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
upstream	NN	O	B-protein
activator	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
is	NN	O	O
unknown	NN	O	O
,	NN	O	O
and	NN	O	O
consequences	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
activation	NN	O	O
remain	NN	O	O
largely	NN	O	O
undefined	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
MAPk	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
MKK	NN	O	B-protein
)	NN	O	O
that	NN	O	O
activates	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
,	NN	O	O
the	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
isoforms	NN	O	I-protein
that	NN	O	O
are	NN	O	O
activated	NN	O	O
as	NN	O	O
part	NN	O	O
of	NN	O	O
this	NN	O	O
pathway	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
functional	NN	O	O
responses	NN	O	O
affected	NN	O	O
by	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Although	NN	O	O
MKK3	NN	O	B-protein
,	NN	O	O
MKK4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
MKK6	NN	O	B-protein
all	NN	O	O
activated	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
in	NN	O	O
experimental	NN	O	O
models	NN	O	O
,	NN	O	O
only	NN	O	O
MKK3	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
activate	NN	O	O
recombinant	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
in	NN	O	O
LPS-treated	NN	O	O
neutrophils	NN	O	O
.	NN	O	O

Of	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
isoforms	NN	O	I-protein
studied	NN	O	O
,	NN	O	O
only	NN	O	O
p38alpha	NN	O	B-protein
and	NN	O	O
p38delta	NN	O	B-protein
were	NN	O	O
detected	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
.	NN	O	O

LPS	NN	O	O
stimulation	NN	O	O
selectively	NN	O	O
activated	NN	O	O
p38alpha	NN	O	B-protein
.	NN	O	O

Specific	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
p38alpha	NN	O	B-protein
MAPk	NN	O	I-protein
blocked	NN	O	O
LPS-induced	NN	O	O
adhesion	NN	O	O
,	NN	O	O
nuclear	NN	O	O
factor-kappa	NN	O	O
B	NN	O	O
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
synthesis	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
p38alpha	NN	O	B-protein
MAPk	NN	O	I-protein
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
transient	NN	O	O
decrease	NN	O	O
in	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
but	NN	O	O
persistent	NN	O	O
loss	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
synthesis	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
support	NN	O	O
a	NN	O	O
pathway	NN	O	O
by	NN	O	O
which	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
results	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
MKK3	NN	O	B-protein
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
activates	NN	O	O
p38alpha	NN	O	B-protein
MAPk	NN	O	I-protein
,	NN	O	O
ultimately	NN	O	O
regulating	NN	O	O
adhesion	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
enhanced	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
synthesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
evolutionarily	NN	O	B-DNA
conserved	NN	O	I-DNA
sequence	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
Ig	NN	O	I-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
S	NN	O	I-protein
gamma	NN	O	I-protein
3	NN	O	I-protein
region	NN	O	I-protein
is	NN	O	O
an	NN	O	O
inducible	NN	O	B-DNA
promoter	NN	O	I-DNA
:	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
ligand	NN	O	I-protein
and	NN	O	O
IL-4	NN	O	B-protein
via	NN	O	O
cooperative	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
STAT-6	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

Germline	NN	O	O
C	NN	O	O
gamma	NN	O	O
gene	NN	O	O
transcription	NN	O	O
is	NN	O	O
a	NN	O	O
crucial	NN	O	O
event	NN	O	O
in	NN	O	O
the	NN	O	O
process	NN	O	O
that	NN	O	O
leads	NN	O	O
to	NN	O	O
switch	NN	O	O
DNA	NN	O	O
recombination	NN	O	O
to	NN	O	O
IgG	NN	O	B-protein
,	NN	O	O
but	NN	O	O
its	NN	O	O
regulation	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
is	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
took	NN	O	O
advantage	NN	O	O
of	NN	O	O
our	NN	O	O
monoclonal	NN	O	O
model	NN	O	O
of	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
B	NN	O	I-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
,	NN	O	O
IgM+	NN	O	B-cell_line
IgD+	NN	O	I-cell_line
CL-01	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
I	NN	O	B-protein
gamma	NN	O	I-protein
3	NN	O	I-protein
evolutionarily	NN	O	B-DNA
conserved	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
ECS	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
germline	NN	O	B-DNA
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
C	NN	O	I-DNA
gamma	NN	O	I-DNA
3	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
I	NN	O	B-protein
gamma	NN	O	I-protein
3	NN	O	I-protein
ECS	NN	O	B-DNA
lies	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
I	NN	O	I-DNA
gamma	NN	O	I-DNA
3	NN	O	I-DNA
transcription	NN	O	I-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
displays	NN	O	O
more	NN	O	O
than	NN	O	O
90	NN	O	O
%	NN	O	O
identity	NN	O	O
with	NN	O	O
the	NN	O	O
corresponding	NN	O	O
human	NN	O	B-DNA
I	NN	O	I-DNA
gamma	NN	O	I-DNA
1	NN	O	I-DNA
,	NN	O	O
I	NN	O	B-DNA
gamma	NN	O	I-DNA
2	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
I	NN	O	B-DNA
gamma	NN	O	I-DNA
4	NN	O	I-DNA
regions	NN	O	O
.	NN	O	O

Reporter	NN	O	B-DNA
luciferase	NN	O	I-DNA
gene	NN	O	I-DNA
vectors	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
gamma	NN	O	I-DNA
3	NN	O	I-DNA
ECS	NN	O	I-DNA
were	NN	O	O
used	NN	O	O
to	NN	O	O
transfect	NN	O	O
CL-01	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
undergo	NN	O	O
Smu	NN	O	O
--	NN	O	O
>	NN	O	O
S	NN	O	O
gamma	NN	O	O
3	NN	O	O
DNA	NN	O	O
recombination	NN	O	O
,	NN	O	O
upon	NN	O	O
engagement	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
by	NN	O	O
CD40	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
CD40L	NN	O	B-protein
)	NN	O	O
and	NN	O	O
exposure	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

In	NN	O	O
these	NN	O	O
transfected	NN	O	O
CL-01	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
CD40	NN	O	O
:	NN	O	O
CD40L	NN	O	B-protein
engagement	NN	O	O
and	NN	O	O
exposure	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
synergistically	NN	O	O
induced	NN	O	O
gamma	NN	O	O
3	NN	O	O
ECS-dependent	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

Targeted	NN	O	O
mutational	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
a	NN	O	O
tandem	NN	O	O
NF-kappa	NN	O	B-DNA
B/Rel	NN	O	I-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
is	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
3	NN	O	I-DNA
ECS	NN	O	I-DNA
responsiveness	NN	O	O
to	NN	O	O
both	NN	O	O
CD40L	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
while	NN	O	O
a	NN	O	O
STAT-6-binding	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
additionally	NN	O	O
required	NN	O	O
for	NN	O	O
IL-4	NN	O	B-protein
inducibility	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
p50/p65/c-Rel	NN	O	B-protein
and	NN	O	O
STAT-6	NN	O	B-protein
are	NN	O	O
effectively	NN	O	O
induced	NN	O	O
by	NN	O	O
CD40L	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
and	NN	O	O
bind	NN	O	O
to	NN	O	O
specific	NN	O	O
DNA	NN	O	B-DNA
motifs	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
ECS	NN	O	B-DNA
.	NN	O	O

These	NN	O	O
partially	NN	O	O
overlapping	NN	O	O
CD40L	NN	O	B-DNA
and	NN	O	I-DNA
IL-4	NN	O	I-DNA
responsive	NN	O	I-DNA
elements	NN	O	I-DNA
are	NN	O	O
functionally	NN	O	O
cooperative	NN	O	O
as	NN	O	O
the	NN	O	O
disruption	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
them	NN	O	O
prevents	NN	O	O
synergistic	NN	O	O
promoter	NN	O	O
activation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
3	NN	O	I-DNA
ECS	NN	O	I-DNA
is	NN	O	O
an	NN	O	O
inducible	NN	O	O
promoter	NN	O	O
containing	NN	O	O
cis	NN	O	B-DNA
elements	NN	O	I-DNA
that	NN	O	O
critically	NN	O	O
mediate	NN	O	O
CD40L	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
-triggered	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
C	NN	O	I-DNA
gamma	NN	O	I-DNA
3	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

CIITA	NN	O	B-protein
-induced	NN	O	O
occupation	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
is	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
cooperative	NN	O	O
stabilization	NN	O	O
of	NN	O	O
the	NN	O	O
promoter-bound	NN	O	B-protein
multi-protein	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Precise	NN	O	O
regulation	NN	O	O
of	NN	O	O
MHC	NN	O	B-cell_type
class	NN	O	I-cell_type
II	NN	O	I-cell_type
expression	NN	O	O
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
transactivator	NN	O	B-protein
CIITA	NN	O	B-protein
behaves	NN	O	O
as	NN	O	O
a	NN	O	O
master	NN	O	O
controller	NN	O	O
of	NN	O	O
constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
activation	NN	O	O
,	NN	O	O
but	NN	O	O
its	NN	O	O
exact	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
requires	NN	O	O
binding	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
distinct	NN	O	O
multi-protein	NN	O	B-protein
complexes	NN	O	I-protein
(	NN	O	O
RFX	NN	O	B-protein
,	NN	O	O
X2BP	NN	O	B-protein
and	NN	O	O
NF-Y	NN	O	B-protein
)	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
known	NN	O	O
that	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
this	NN	O	O
binding	NN	O	O
results	NN	O	O
from	NN	O	O
cooperative	NN	O	O
interactions	NN	O	O
between	NN	O	O
these	NN	O	O
proteins	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
in	NN	O	O
MHC	NN	O	B-cell_line
class	NN	O	I-cell_line
II	NN	O	I-cell_line
-cells	NN	O	I-cell_line
triggers	NN	O	O
occupation	NN	O	O
of	NN	O	O
the	NN	O	O
promoters	NN	O	O
by	NN	O	O
these	NN	O	O
complexes	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
raised	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
on	NN	O	O
promoter	NN	O	O
occupation	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
an	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
cooperative	NN	O	O
stabilization	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-bound	NN	O	B-protein
multi-protein	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
does	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
the	NN	O	O
higher-order	NN	O	O
protein	NN	O	O
complex	NN	O	O
formed	NN	O	O
on	NN	O	O
DNA	NN	O	O
by	NN	O	O
RFX	NN	O	B-protein
,	NN	O	O
X2BP	NN	O	B-protein
and	NN	O	O
NF-Y	NN	O	B-protein
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
other	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
CIITA	NN	O	B-protein
-induced	NN	O	O
promoter	NN	O	O
occupancy	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
an	NN	O	O
effect	NN	O	O
on	NN	O	O
chromatin	NN	O	B-DNA
structure	NN	O	O
leading	NN	O	O
to	NN	O	O
increased	NN	O	O
accessibility	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
ability	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
to	NN	O	O
facilitate	NN	O	O
promoter	NN	O	O
occupation	NN	O	O
is	NN	O	O
undissociable	NN	O	O
from	NN	O	O
its	NN	O	O
transactivation	NN	O	O
potential	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
this	NN	O	O
effect	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
is	NN	O	O
cell-type	NN	O	O
specific	NN	O	O
,	NN	O	O
since	NN	O	O
expression	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
normal	NN	O	O
occupation	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

GrpL	NN	O	B-protein
,	NN	O	O
a	NN	O	O
Grb2	NN	O	B-protein
-related	NN	O	O
adaptor	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
interacts	NN	O	O
with	NN	O	O
SLP-76	NN	O	B-protein
to	NN	O	O
regulate	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Propagation	NN	O	O
of	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
involves	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
adaptor	NN	O	O
molecules	NN	O	O
.	NN	O	O

SH2	NN	O	B-protein
domain-containing	NN	O	I-protein
protein	NN	O	I-protein
76	NN	O	I-protein
(	NN	O	O
SLP-76	NN	O	B-protein
)	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
guanine	NN	O	B-protein
nucleotide	NN	O	I-protein
exchange	NN	O	I-protein
factor	NN	O	I-protein
Vav	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
expression	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
development	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
cloning	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
Grb2-like	NN	O	B-protein
adaptor	NN	O	I-protein
molecule	NN	O	I-protein
designated	NN	O	O
as	NN	O	O
Grb2-related	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	I-protein
the	NN	O	I-protein
lymphoid	NN	O	I-protein
system	NN	O	I-protein
(	NN	O	O
GrpL	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
GrpL	NN	O	B-protein
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
hematopoietic	NN	O	O
tissues	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
is	NN	O	O
distinguished	NN	O	O
from	NN	O	O
Grb2	NN	O	B-protein
by	NN	O	O
having	NN	O	O
a	NN	O	O
proline-rich	NN	O	B-protein
region	NN	O	I-protein
.	NN	O	O

GrpL	NN	O	B-protein
can	NN	O	O
be	NN	O	O
coimmunoprecipitated	NN	O	O
with	NN	O	O
SLP-76	NN	O	B-protein
but	NN	O	O
not	NN	O	O
with	NN	O	O
Sos1	NN	O	B-protein
or	NN	O	O
Sos2	NN	O	B-protein
from	NN	O	O
Jurkat	NN	O	B-cell_line
cell	NN	O	I-cell_line
lysates	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
Grb2	NN	O	B-protein
can	NN	O	O
be	NN	O	O
coimmunoprecipitated	NN	O	O
with	NN	O	O
Sos1	NN	O	B-protein
and	NN	O	O
Sos2	NN	O	B-protein
but	NN	O	O
not	NN	O	O
with	NN	O	O
SLP-76	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
tyrosine-phosphorylated	NN	O	B-protein
LAT/pp36/38	NN	O	I-protein
in	NN	O	O
detergent	NN	O	O
lysates	NN	O	O
prepared	NN	O	O
from	NN	O	O
anti-CD3	NN	O	B-cell_type
stimulated	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
associated	NN	O	O
with	NN	O	O
Grb2	NN	O	B-protein
but	NN	O	O
not	NN	O	O
GrpL	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
reveal	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
distinct	NN	O	O
complexes	NN	O	O
involving	NN	O	O
GrpL	NN	O	B-protein
and	NN	O	O
Grb2	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
functional	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
GrpL-SLP-76	NN	O	B-protein
complex	NN	O	I-protein
is	NN	O	O
suggested	NN	O	O
by	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
GrpL	NN	O	B-protein
to	NN	O	O
act	NN	O	O
alone	NN	O	O
or	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
SLP-76	NN	O	B-protein
to	NN	O	O
augment	NN	O	O
NF-AT	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
induction	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
(	NN	O	I-protein
IFN	NN	O	I-protein
)	NN	O	I-protein
-inducible	NN	O	I-protein
protein	NN	O	I-protein
10	NN	O	I-protein
following	NN	O	O
differentiation	NN	O	O
of	NN	O	O
a	NN	O	O
monocyte	NN	O	B-cell_type
,	NN	O	O
macrophage	NN	O	B-cell_type
cell	NN	O	I-cell_type
lineage	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
changes	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
IFN	NN	O	B-DNA
stimulus	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
and	NN	O	O
kappaB	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
examined	NN	O	O
chemokine	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
following	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
a	NN	O	O
monocyte	NN	O	B-cell_type
,	NN	O	O
macrophage	NN	O	B-cell_line
cell	NN	O	I-cell_line
lineage	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
human	NN	O	B-cell_line
monoblastic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
U937	NN	O	B-cell_line
was	NN	O	O
differentiated	NN	O	O
to	NN	O	O
macrophages	NN	O	B-cell_type
by	NN	O	O
the	NN	O	O
treatment	NN	O	O
with	NN	O	O
either	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
,	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
VitD3	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
(	NN	O	I-protein
IFN	NN	O	I-protein
)	NN	O	I-protein
-inducible	NN	O	I-protein
protein	NN	O	I-protein
10	NN	O	I-protein
(	NN	O	O
IP-10	NN	O	B-protein
)	NN	O	O
(	NN	O	O
a	NN	O	O
CXC	NN	O	B-protein
chemokine	NN	O	I-protein
)	NN	O	O
was	NN	O	O
markedly	NN	O	O
augmented	NN	O	O
by	NN	O	O
the	NN	O	O
IFNgamma	NN	O	B-protein
treatment	NN	O	O
in	NN	O	O
PMA	NN	O	O
-or	NN	O	O
RA-differentiated	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
only	NN	O	O
marginally	NN	O	O
in	NN	O	O
undifferentiated	NN	O	O
or	NN	O	O
VitD3-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
another	NN	O	O
inducible	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
monocyte	NN	O	B-protein
chemotactic	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
a	NN	O	O
CC	NN	O	B-protein
chemokine	NN	O	I-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
FcRFgamma	NN	O	B-protein
)	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
response	NN	O	I-DNA
region	NN	O	I-DNA
were	NN	O	O
similarly	NN	O	O
or	NN	O	O
less	NN	O	O
abundantly	NN	O	O
induced	NN	O	O
by	NN	O	O
IFNgamma	NN	O	B-protein
treatment	NN	O	O
in	NN	O	O
PMA	NN	O	O
-or	NN	O	O
RA-differentiated	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
increased	NN	O	O
IP-10	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
was	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
augmented	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IFNgamma	NN	O	B-protein
.	NN	O	O

Increased	NN	O	O
expression	NN	O	O
of	NN	O	O
IFNgamma	NN	O	B-protein
-induced	NN	O	O
IP-10	NN	O	B-RNA
mRNA	NN	O	I-RNA
following	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
mediated	NN	O	O
largely	NN	O	O
by	NN	O	O
augmented	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
and	NN	O	O
was	NN	O	O
related	NN	O	O
to	NN	O	O
differentiation-dependent	NN	O	O
changes	NN	O	O
of	NN	O	O
the	NN	O	O
proteins	NN	O	O
bound	NN	O	O
to	NN	O	O
IFN	NN	O	B-DNA
stimulus	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
ISRE	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
kB	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
may	NN	O	O
determine	NN	O	O
the	NN	O	O
IP-10	NN	O	B-RNA
mRNA	NN	O	I-RNA
inducibility	NN	O	O
by	NN	O	O
IFNgamma	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Role	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-associated	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
lymphocyte	NN	O	B-cell_type
transformation	NN	O	O
by	NN	O	O
herpesvirus	NN	O	O
Saimiri	NN	O	O
STP	NN	O	B-protein
.	NN	O	O

The	NN	O	O
STP	NN	O	B-protein
oncoproteins	NN	O	I-protein
of	NN	O	O
the	NN	O	O
herpesvirus	NN	O	O
saimiri	NN	O	O
(	NN	O	O
HVS	NN	O	O
)	NN	O	O
subgroup	NN	O	O
A	NN	O	O
strain	NN	O	O
11	NN	O	O
and	NN	O	O
subgroup	NN	O	O
C	NN	O	O
strain	NN	O	O
488	NN	O	O
are	NN	O	O
now	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
stably	NN	O	O
associated	NN	O	O
with	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-associated	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TRAF	NN	O	I-protein
)	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	I-protein
2	NN	O	I-protein
,	NN	O	I-protein
or	NN	O	I-protein
3	NN	O	I-protein
.	NN	O	O

Mutational	NN	O	O
analyses	NN	O	O
identified	NN	O	O
residues	NN	O	O
of	NN	O	O
PXQXT/S	NN	O	B-protein
in	NN	O	O
STP-A11	NN	O	B-protein
as	NN	O	O
critical	NN	O	O
for	NN	O	O
TRAF	NN	O	B-protein
association	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
a	NN	O	O
somewhat	NN	O	O
divergent	NN	O	O
region	NN	O	O
of	NN	O	O
STP-C488	NN	O	B-protein
is	NN	O	O
critical	NN	O	O
for	NN	O	O
TRAF	NN	O	B-protein
association	NN	O	O
.	NN	O	O

Mutational	NN	O	O
analysis	NN	O	O
also	NN	O	O
revealed	NN	O	O
that	NN	O	O
STP-C488	NN	O	B-protein
induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
that	NN	O	O
was	NN	O	O
correlated	NN	O	O
with	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
associate	NN	O	O
with	NN	O	O
TRAFs	NN	O	B-protein
.	NN	O	O

The	NN	O	O
HVS	NN	O	O
STP-C488	NN	O	B-protein
P10	NN	O	O
--	NN	O	O
>	NN	O	O
R	NN	O	O
mutant	NN	O	O
was	NN	O	O
deficient	NN	O	O
in	NN	O	O
human	NN	O	O
T-lymphocyte	NN	O	O
transformation	NN	O	O
to	NN	O	O
interleukin-2-independent	NN	O	O
growth	NN	O	O
but	NN	O	O
showed	NN	O	O
wild-type	NN	O	O
phenotype	NN	O	O
for	NN	O	O
marmoset	NN	O	O
T-lymphocyte	NN	O	O
transformation	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

The	NN	O	O
STP-C488	NN	O	B-protein
P10	NN	O	I-protein
--	NN	O	I-protein
>	NN	O	I-protein
R	NN	O	I-protein
mutant	NN	O	I-protein
was	NN	O	O
also	NN	O	O
defective	NN	O	O
in	NN	O	O
Rat-1	NN	O	O
fibroblast	NN	O	O
transformation	NN	O	O
,	NN	O	O
and	NN	O	O
fibroblast	NN	O	O
cell	NN	O	O
transformation	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
a	NN	O	O
TRAF2	NN	O	B-protein
dominant-negative	NN	O	O
mutant	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
implicate	NN	O	O
TRAFs	NN	O	B-protein
in	NN	O	O
STP-C488	NN	O	B-protein
-mediated	NN	O	O
transformation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
rodent	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

Other	NN	O	O
factors	NN	O	O
are	NN	O	O
implicated	NN	O	O
in	NN	O	O
immortalization	NN	O	O
of	NN	O	O
common	NN	O	B-cell_type
marmoset	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
may	NN	O	O
also	NN	O	O
be	NN	O	O
critical	NN	O	O
in	NN	O	O
the	NN	O	O
transformation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
rodent	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Cleavage	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
SP1	NN	O	B-protein
by	NN	O	O
caspases	NN	O	B-protein
during	NN	O	O
anti-IgM-induced	NN	O	O
B-cell	NN	O	B-cell_type
apoptosis	NN	O	O
.	NN	O	O

Apoptosis	NN	O	O
is	NN	O	O
instrumental	NN	O	O
in	NN	O	O
the	NN	O	O
processes	NN	O	O
generating	NN	O	O
the	NN	O	O
diversity	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell	NN	O	O
repertoire	NN	O	O
.	NN	O	O

Autoreactive	NN	O	B-cell_type
B-cells	NN	O	I-cell_type
are	NN	O	O
eliminated	NN	O	O
by	NN	O	O
anti-IgM	NN	O	B-protein
crosslinking	NN	O	O
after	NN	O	O
encountering	NN	O	O
self-antigens	NN	O	B-protein
,	NN	O	O
but	NN	O	O
precise	NN	O	O
mechanisms	NN	O	O
leading	NN	O	O
to	NN	O	O
B-cell	NN	O	B-cell_type
apoptosis	NN	O	O
are	NN	O	O
still	NN	O	O
not	NN	O	O
well	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
cleavage	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
SP1	NN	O	B-protein
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
Burkitt	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
BL60	NN	O	I-cell_line
during	NN	O	O
anti-IgM	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
two	NN	O	O
cleavage	NN	O	O
products	NN	O	O
of	NN	O	O
approximately	NN	O	O
68	NN	O	B-protein
kDa	NN	O	I-protein
and	NN	O	O
45	NN	O	B-protein
kDa	NN	O	I-protein
after	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Cleavage	NN	O	O
could	NN	O	O
be	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
by	NN	O	O
zDEVD-fmk	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
specific	NN	O	O
for	NN	O	O
caspase	NN	O	B-protein
3-like	NN	O	I-protein
proteases	NN	O	I-protein
.	NN	O	O

In-vitro	NN	O	O
cleavage	NN	O	O
of	NN	O	O
recombinant	NN	O	B-protein
SP1	NN	O	I-protein
by	NN	O	O
recombinant	NN	O	B-protein
caspase	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
CPP32	NN	O	B-protein
)	NN	O	O
or	NN	O	O
caspase	NN	O	B-protein
7	NN	O	I-protein
(	NN	O	O
Mch	NN	O	B-protein
3	NN	O	I-protein
)	NN	O	O
results	NN	O	O
in	NN	O	O
similar	NN	O	O
cleavage	NN	O	O
products	NN	O	O
as	NN	O	O
those	NN	O	O
observed	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Recombinant	NN	O	B-protein
caspase	NN	O	I-protein
6	NN	O	I-protein
(	NN	O	O
Mch	NN	O	B-protein
2	NN	O	I-protein
)	NN	O	O
primarily	NN	O	O
generates	NN	O	O
a	NN	O	O
68-kDa	NN	O	O
cleavage	NN	O	O
product	NN	O	O
,	NN	O	O
as	NN	O	O
observed	NN	O	O
after	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
(	NN	O	O
CaI	NN	O	O
)	NN	O	O
induced	NN	O	O
B-cell	NN	O	B-cell_type
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
caspase	NN	O	O
1	NN	O	O
(	NN	O	O
ICE	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
cleave	NN	O	O
SP1	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
time	NN	O	O
course	NN	O	O
of	NN	O	O
SP1	NN	O	B-protein
cleavage	NN	O	O
during	NN	O	O
anti-IgM-induced	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
paralleled	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
of	NN	O	O
caspase	NN	O	O
activity	NN	O	O
measured	NN	O	O
by	NN	O	O
DEVD-p-nitroanilide	NN	O	O
(	NN	O	O
DEVD-pNA	NN	O	O
)	NN	O	O
cleavage	NN	O	O
.	NN	O	O

DNA	NN	O	O
band-shift	NN	O	O
assays	NN	O	O
revealed	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
intensity	NN	O	O
of	NN	O	O
the	NN	O	O
full	NN	O	O
length	NN	O	O
SP1	NN	O	B-protein
/DNA	NN	O	O
complex	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
intensity	NN	O	O
of	NN	O	O
a	NN	O	O
smaller	NN	O	O
complex	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
one	NN	O	O
SP1	NN	O	B-protein
cleavage	NN	O	O
product	NN	O	O
.	NN	O	O

By	NN	O	O
Edman	NN	O	O
sequencing	NN	O	O
we	NN	O	O
could	NN	O	O
identify	NN	O	O
a	NN	O	O
caspase	NN	O	O
3	NN	O	O
cleavage	NN	O	O
site	NN	O	O
after	NN	O	O
Asp584	NN	O	O
(	NN	O	O
D584AQPQAGR	NN	O	O
)	NN	O	O
,	NN	O	O
generating	NN	O	O
a	NN	O	O
22-kDa	NN	O	O
C-terminal	NN	O	O
SP1	NN	O	B-protein
protein	NN	O	O
fragment	NN	O	O
which	NN	O	O
still	NN	O	O
contains	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
site	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
show	NN	O	O
the	NN	O	O
cleavage	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
SP1	NN	O	B-protein
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
underlining	NN	O	O
the	NN	O	O
central	NN	O	O
role	NN	O	O
of	NN	O	O
caspase	NN	O	B-protein
3-like	NN	O	I-protein
proteases	NN	O	I-protein
during	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
anti-IgM-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stimulation	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
on	NN	O	O
immunogenic	NN	O	O
human	NN	O	O
malignant	NN	O	O
melanomas	NN	O	O
augments	NN	O	O
their	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
-mediated	NN	O	O
lysis	NN	O	O
and	NN	O	O
induces	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
functional	NN	O	O
expression	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
on	NN	O	O
human	NN	O	O
malignant	NN	O	O
melanomas	NN	O	O
(	NN	O	O
MMs	NN	O	O
)	NN	O	O
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
tumor	NN	O	O
specimen	NN	O	O
from	NN	O	O
MM	NN	O	O
precursor	NN	O	O
lesions	NN	O	O
,	NN	O	O
primary	NN	O	O
tumors	NN	O	O
,	NN	O	O
and	NN	O	O
metastases	NN	O	O
revealed	NN	O	O
that	NN	O	O
CD40	NN	O	B-protein
surface	NN	O	O
expression	NN	O	O
is	NN	O	O
down-regulated	NN	O	O
during	NN	O	O
tumor	NN	O	O
progression	NN	O	O
.	NN	O	O

CD40	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
confirmed	NN	O	O
in	NN	O	O
7	NN	O	O
human	NN	O	B-cell_line
MM	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
established	NN	O	O
from	NN	O	O
immunogenic	NN	O	O
primary	NN	O	O
tumors	NN	O	O
or	NN	O	O
metastases	NN	O	O
,	NN	O	O
whereas	NN	O	O
11	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
established	NN	O	O
from	NN	O	O
advanced	NN	O	O
stages	NN	O	O
were	NN	O	O
CD40	NN	O	B-protein
negative	NN	O	O
.	NN	O	O

CD40	NN	O	B-protein
expression	NN	O	O
could	NN	O	O
be	NN	O	O
enhanced	NN	O	O
in	NN	O	O
CD40	NN	O	B-protein
-positive	NN	O	O
MM	NN	O	O
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
but	NN	O	O
not	NN	O	O
by	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-1beta	NN	O	I-protein
or	NN	O	O
CD40	NN	O	B-protein
triggering	NN	O	O
.	NN	O	O

CD40	NN	O	B-protein
ligation	NN	O	O
on	NN	O	O
MM	NN	O	O
by	NN	O	O
CD40L-transfected	NN	O	B-cell_line
murine	NN	O	I-cell_line
L-cells	NN	O	I-cell_line
or	NN	O	O
by	NN	O	O
a	NN	O	O
soluble	NN	O	O
CD40L	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
up-regulated	NN	O	O
their	NN	O	O
expression	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
and	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
and	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
and	NN	O	O
their	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-a	NN	O	I-protein
,	NN	O	O
and	NN	O	O
granulocyte	NN	O	B-protein
macrophage	NN	O	I-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
also	NN	O	O
induced	NN	O	O
a	NN	O	O
rapid	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
CD40	NN	O	B-protein
ligation	NN	O	O
of	NN	O	O
a	NN	O	O
HLA-A2+	NN	O	B-cell_line
,	NN	O	I-cell_line
MelanA/MART1+	NN	O	I-cell_line
MM	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
enhanced	NN	O	O
its	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
specific	NN	O	O
lysis	NN	O	O
by	NN	O	O
a	NN	O	O
HLA-A2-restricted	NN	O	B-cell_line
,	NN	O	I-cell_line
MelanA/MART-1-specific	NN	O	I-cell_line
CTL	NN	O	I-cell_line
clone	NN	O	I-cell_line
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
CD40	NN	O	B-protein
ligation	NN	O	O
induced	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
MM	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
CD40	NN	O	B-protein
-CD40L	NN	O	B-protein
interactions	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
augmenting	NN	O	O
antitumor	NN	O	O
immunity	NN	O	O
and	NN	O	O
inducing	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
some	NN	O	O
CD40	NN	O	B-protein
-positive	NN	O	O
immunogenic	NN	O	O
human	NN	O	O
MMs	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus-associated	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
derives	NN	O	O
from	NN	O	O
post-germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus-associated	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
(	NN	O	O
HIV-HD	NN	O	O
)	NN	O	O
displays	NN	O	O
several	NN	O	O
peculiarities	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
HD	NN	O	O
of	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
.	NN	O	O

These	NN	O	O
include	NN	O	O
overrepresentation	NN	O	O
of	NN	O	O
clinically	NN	O	O
aggressive	NN	O	O
histologic	NN	O	O
types	NN	O	O
and	NN	O	O
frequent	NN	O	O
infection	NN	O	O
of	NN	O	O
Reed-Sternberg	NN	O	B-cell_line
(	NN	O	I-cell_line
RS	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
reported	NN	O	O
that	NN	O	O
the	NN	O	O
histogenesis	NN	O	O
of	NN	O	O
HD	NN	O	O
of	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
may	NN	O	O
be	NN	O	O
assessed	NN	O	O
by	NN	O	O
monitoring	NN	O	O
the	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
germinal	NN	O	B-cell_line
center	NN	O	I-cell_line
(	NN	O	I-cell_line
GC	NN	O	I-cell_line
)	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
of	NN	O	O
CD138/syndecan-1	NN	O	B-protein
(	NN	O	O
syn-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
proteoglycan	NN	O	O
associated	NN	O	O
with	NN	O	O
post-GC	NN	O	O
,	NN	O	O
terminal	NN	O	O
B-cell	NN	O	B-cell_type
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
applied	NN	O	O
these	NN	O	O
two	NN	O	O
markers	NN	O	O
to	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
HIV-HD	NN	O	O
histogenesis	NN	O	O
and	NN	O	O
correlated	NN	O	O
their	NN	O	O
expression	NN	O	O
status	NN	O	O
to	NN	O	O
the	NN	O	O
virologic	NN	O	O
features	NN	O	O
of	NN	O	O
this	NN	O	O
disease	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
RS	NN	O	B-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
all	NN	O	O
histologic	NN	O	O
categories	NN	O	O
of	NN	O	O
HIV-HD	NN	O	O
consistently	NN	O	O
display	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
(	NN	O	O
-	NN	O	O
)	NN	O	O
/syn-1	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
phenotype	NN	O	O
and	NN	O	O
thus	NN	O	O
reflect	NN	O	O
post-GC	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Although	NN	O	O
BCL-6	NN	O	B-protein
(	NN	O	O
-	NN	O	O
)	NN	O	O
/syn-1	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
RS	NN	O	B-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
HIV-HD	NN	O	O
express	NN	O	O
CD40	NN	O	B-protein
,	NN	O	O
they	NN	O	O
are	NN	O	O
not	NN	O	O
surrounded	NN	O	O
by	NN	O	O
CD40	NN	O	B-cell_type
ligand-positive	NN	O	I-cell_type
(	NN	O	I-cell_type
CD40L+	NN	O	I-cell_type
)	NN	O	I-cell_type
reactive	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
,	NN	O	O
in	NN	O	O
HD	NN	O	O
of	NN	O	O
the	NN	O	O
general	NN	O	O
population	NN	O	O
,	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
regulate	NN	O	O
the	NN	O	O
disease	NN	O	O
phenotype	NN	O	O
through	NN	O	O
CD40	NN	O	B-protein
/CD40L	NN	O	B-protein
interactions	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
RS	NN	O	B-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
virtually	NN	O	O
all	NN	O	O
HIV-HD	NN	O	O
express	NN	O	O
the	NN	O	O
EBV-encoded	NN	O	B-protein
latent	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
LMP1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
,	NN	O	O
being	NN	O	O
functionally	NN	O	O
homologous	NN	O	O
to	NN	O	O
CD40	NN	O	B-protein
,	NN	O	O
may	NN	O	O
contribute	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
to	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-HD	NN	O	O
phenotype	NN	O	O
.	NN	O	O

-DOCSTART-	O

MDS1/EVI1	NN	O	B-DNA
enhances	NN	O	O
TGF-beta1	NN	O	B-protein
signaling	NN	O	O
and	NN	O	O
strengthens	NN	O	O
its	NN	O	O
growth-inhibitory	NN	O	O
effect	NN	O	O
but	NN	O	O
the	NN	O	O
leukemia-associated	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
AML1/MDS1/EVI1	NN	O	B-protein
,	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
t	NN	O	O
(	NN	O	O
3	NN	O	O
;	NN	O	O
21	NN	O	O
)	NN	O	O
,	NN	O	O
abrogates	NN	O	O
growth-inhibition	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TGF-beta1	NN	O	B-protein
.	NN	O	O

MDS1/EVI1	NN	O	B-DNA
,	NN	O	O
located	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
3	NN	O	I-DNA
band	NN	O	I-DNA
q26	NN	O	I-DNA
,	NN	O	O
encodes	NN	O	O
a	NN	O	O
zinc-finger	NN	O	B-protein
DNA-binding	NN	O	I-protein
transcription	NN	O	I-protein
activator	NN	O	I-protein
not	NN	O	O
detected	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
expressed	NN	O	O
in	NN	O	O
several	NN	O	O
normal	NN	O	O
tissues	NN	O	O
.	NN	O	O

MDS1/EVI1	NN	O	B-DNA
is	NN	O	O
inappropriately	NN	O	O
activated	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
leukemias	NN	O	I-cell_type
following	NN	O	O
chromosomal	NN	O	O
rearrangements	NN	O	O
involving	NN	O	O
band	NN	O	B-protein
3q26	NN	O	I-protein
.	NN	O	O

The	NN	O	O
rearrangements	NN	O	O
lead	NN	O	O
either	NN	O	O
to	NN	O	O
gene	NN	O	O
truncation	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
repressor	NN	O	I-protein
EVI1	NN	O	I-protein
,	NN	O	O
as	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
3	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q21	NN	O	I-DNA
;	NN	O	I-DNA
q26	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
inv	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q21q26	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
or	NN	O	O
to	NN	O	O
gene	NN	O	O
fusion	NN	O	O
,	NN	O	O
as	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
21	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q26	NN	O	I-DNA
;	NN	O	I-DNA
q22	NN	O	I-DNA
)	NN	O	I-DNA
which	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
fusion	NN	O	O
protein	NN	O	O
AML1/MDS1/EVI1	NN	O	B-protein
.	NN	O	O

This	NN	O	O
fusion	NN	O	O
protein	NN	O	O
contains	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AML1	NN	O	B-protein
fused	NN	O	O
in-frame	NN	O	O
to	NN	O	O
the	NN	O	O
entire	NN	O	O
MDS1/EVI1	NN	O	B-DNA
with	NN	O	O
the	NN	O	O
exclusion	NN	O	O
of	NN	O	O
its	NN	O	O
first	NN	O	O
12	NN	O	O
amino	NN	O	O
acids	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-cell_line
precursor	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
32Dcl3	NN	O	I-cell_line
,	NN	O	O
expressing	NN	O	O
either	NN	O	O
the	NN	O	O
normal	NN	O	B-protein
protein	NN	O	I-protein
MDS1/EVI1	NN	O	I-protein
or	NN	O	O
the	NN	O	O
fusion	NN	O	B-protein
protein	NN	O	I-protein
AML1/MDS1/EVI1	NN	O	I-protein
,	NN	O	O
to	NN	O	O
factors	NN	O	O
that	NN	O	O
control	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
or	NN	O	O
cell	NN	O	O
replication	NN	O	O
.	NN	O	O

The	NN	O	O
32Dcl3	NN	O	B-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
IL-3	NN	O	B-protein
-dependent	NN	O	O
for	NN	O	O
growth	NN	O	O
and	NN	O	O
they	NN	O	O
differentiate	NN	O	O
into	NN	O	O
granulocytes	NN	O	O
when	NN	O	O
exposed	NN	O	O
to	NN	O	O
G-CSF	NN	O	B-protein
.	NN	O	O

They	NN	O	O
are	NN	O	O
growth-inhibited	NN	O	O
by	NN	O	O
TGF-beta1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
whereas	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
MDS1/EVI1	NN	O	B-DNA
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
G-CSF	NN	O	B-protein
,	NN	O	O
expression	NN	O	O
of	NN	O	O
AML1/MDS1/EVI1	NN	O	B-protein
blocks	NN	O	O
differentiation	NN	O	O
resulting	NN	O	O
in	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

This	NN	O	O
effect	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
previously	NN	O	O
described	NN	O	O
by	NN	O	O
others	NN	O	O
for	NN	O	O
32Dcl3	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
express	NN	O	O
transgenic	NN	O	B-protein
Evil	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
whereas	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
fusion	NN	O	O
protein	NN	O	O
AML1/MDS1/EVI1	NN	O	B-protein
completely	NN	O	O
abrogates	NN	O	O
the	NN	O	O
growth-inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
TGF-beta1	NN	O	B-protein
and	NN	O	O
allows	NN	O	O
32Dcl3	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
proliferate	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
protein	NN	O	O
MDS1/EVI1	NN	O	B-DNA
has	NN	O	O
the	NN	O	O
opposite	NN	O	O
effect	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
strengthens	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
cells	NN	O	O
to	NN	O	O
the	NN	O	O
growth-inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
TGF-beta1	NN	O	B-protein
.	NN	O	O

By	NN	O	O
using	NN	O	O
the	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
system	NN	O	O
,	NN	O	O
we	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
EVI1	NN	O	B-protein
(	NN	O	O
contained	NN	O	O
in	NN	O	O
its	NN	O	O
entirety	NN	O	O
in	NN	O	O
MDS1/EVI1	NN	O	B-DNA
and	NN	O	O
AML1/MDS1/EVI1	NN	O	B-protein
)	NN	O	O
physically	NN	O	O
interacts	NN	O	O
with	NN	O	O
SMAD3	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
an	NN	O	O
intracellular	NN	O	O
mediator	NN	O	O
of	NN	O	O
TGF-beta1	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
correlated	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
to	NN	O	O
G-CSF	NN	O	B-protein
or	NN	O	O
TGF-beta1	NN	O	B-protein
with	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
and	NN	O	O
fusion	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
activate	NN	O	O
or	NN	O	O
repress	NN	O	O
promoters	NN	O	O
which	NN	O	O
they	NN	O	O
can	NN	O	O
directly	NN	O	O
regulate	NN	O	O
by	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
mutations	NN	O	O
of	NN	O	O
MDS1/EVI1	NN	O	B-DNA
either	NN	O	O
by	NN	O	O
gene	NN	O	O
truncation	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
repressor	NN	O	I-protein
EVI1	NN	O	I-protein
or	NN	O	O
by	NN	O	O
gene	NN	O	O
fusion	NN	O	O
to	NN	O	O
AML1	NN	O	B-protein
lead	NN	O	O
to	NN	O	O
an	NN	O	O
altered	NN	O	O
cellular	NN	O	O
response	NN	O	O
to	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
could	NN	O	O
result	NN	O	O
in	NN	O	O
leukemic	NN	O	O
transformation	NN	O	O
.	NN	O	O

The	NN	O	O
different	NN	O	O
response	NN	O	O
of	NN	O	O
myeloid	NN	O	O
cells	NN	O	O
ectopically	NN	O	O
expressing	NN	O	O
the	NN	O	O
normal	NN	O	O
or	NN	O	O
the	NN	O	O
fusion	NN	O	B-protein
protein	NN	O	I-protein
to	NN	O	O
G-CSF	NN	O	B-protein
and	NN	O	O
TGF-beta1	NN	O	B-protein
could	NN	O	O
depend	NN	O	O
on	NN	O	O
the	NN	O	O
different	NN	O	O
transactivation	NN	O	O
properties	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
resulting	NN	O	O
in	NN	O	O
divergent	NN	O	O
expression	NN	O	O
of	NN	O	O
downstream	NN	O	B-DNA
genes	NN	O	I-DNA
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
two	NN	O	O
proteins	NN	O	O
.	NN	O	O

-DOCSTART-	O

Abnormalities	NN	O	O
of	NN	O	O
cyclic	NN	O	O
adenosine	NN	O	O
monophosphate	NN	O	O
signaling	NN	O	O
in	NN	O	O
platelets	NN	O	B-cell_type
from	NN	O	O
untreated	NN	O	O
patients	NN	O	O
with	NN	O	O
bipolar	NN	O	O
disorder	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Abnormalities	NN	O	O
in	NN	O	O
the	NN	O	O
cyclic	NN	O	O
adenosine	NN	O	O
monophosphate	NN	O	O
(	NN	O	O
cAMP	NN	O	O
)	NN	O	O
-dependent	NN	O	O
phosphorylation	NN	O	O
system	NN	O	O
have	NN	O	O
been	NN	O	O
recently	NN	O	O
reported	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
bipolar	NN	O	O
disorder	NN	O	O
.	NN	O	O

We	NN	O	O
evaluated	NN	O	O
the	NN	O	O
immunoreactivity	NN	O	O
of	NN	O	O
the	NN	O	O
regulatory	NN	O	B-protein
and	NN	O	I-protein
catalytic	NN	O	I-protein
subunits	NN	O	I-protein
of	NN	O	O
cAMP-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	O
and	NN	O	O
1	NN	O	O
of	NN	O	O
its	NN	O	O
substrates	NN	O	O
,	NN	O	O
Rap1	NN	O	B-protein
,	NN	O	O
in	NN	O	O
platelets	NN	O	B-cell_type
from	NN	O	O
untreated	NN	O	O
euthymic	NN	O	O
,	NN	O	O
manic	NN	O	O
,	NN	O	O
and	NN	O	O
depressed	NN	O	O
patients	NN	O	O
with	NN	O	O
bipolar	NN	O	O
disorder	NN	O	O
and	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Platelets	NN	O	B-cell_type
were	NN	O	O
collected	NN	O	O
from	NN	O	O
112	NN	O	O
drug-free	NN	O	O
patients	NN	O	O
with	NN	O	O
bipolar	NN	O	O
disorder	NN	O	O
(	NN	O	O
52	NN	O	O
euthymic	NN	O	O
,	NN	O	O
29	NN	O	O
depressed	NN	O	O
,	NN	O	O
and	NN	O	O
31	NN	O	O
manic	NN	O	O
)	NN	O	O
and	NN	O	O
62	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
levels	NN	O	O
of	NN	O	O
cAMP-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
Rap1	NN	O	B-protein
were	NN	O	O
assessed	NN	O	O
by	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
immunostaining	NN	O	O
,	NN	O	O
and	NN	O	O
computer-assisted	NN	O	O
imaging	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
The	NN	O	O
immunolabeling	NN	O	O
of	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
cAMP-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
was	NN	O	O
significantly	NN	O	O
different	NN	O	O
among	NN	O	O
groups	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
.001	NN	O	O
)	NN	O	O
,	NN	O	O
with	NN	O	O
higher	NN	O	O
values	NN	O	O
in	NN	O	O
untreated	NN	O	O
depressed	NN	O	O
and	NN	O	O
manic	NN	O	O
patients	NN	O	O
with	NN	O	O
bipolar	NN	O	O
disorder	NN	O	O
compared	NN	O	O
with	NN	O	O
untreated	NN	O	O
euthymic	NN	O	O
patients	NN	O	O
with	NN	O	O
bipolar	NN	O	O
disorder	NN	O	O
and	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
.	NN	O	O

No	NN	O	O
significant	NN	O	O
differences	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
immunolabeling	NN	O	O
of	NN	O	O
the	NN	O	O
regulatory	NN	O	B-protein
subunits	NN	O	I-protein
(	NN	O	O
type	NN	O	O
I	NN	O	O
and	NN	O	O
type	NN	O	O
II	NN	O	O
)	NN	O	O
of	NN	O	O
cAMP-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

The	NN	O	O
immunolabeling	NN	O	O
of	NN	O	O
Rap1	NN	O	B-protein
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
.001	NN	O	O
)	NN	O	O
in	NN	O	O
untreated	NN	O	O
euthymic	NN	O	O
,	NN	O	O
depressed	NN	O	O
,	NN	O	O
and	NN	O	O
manic	NN	O	O
patients	NN	O	O
than	NN	O	O
in	NN	O	O
healthy	NN	O	O
persons	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Levels	NN	O	O
of	NN	O	O
Rap1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
cAMP-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
are	NN	O	O
altered	NN	O	O
in	NN	O	O
the	NN	O	O
platelets	NN	O	B-cell_type
of	NN	O	O
bipolar	NN	O	O
patients	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
may	NN	O	O
provide	NN	O	O
clues	NN	O	O
toward	NN	O	O
understanding	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
cAMP	NN	O	O
signaling	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
bipolar	NN	O	O
disorder	NN	O	O
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Cytokine	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
immune	NN	O	O
system	NN	O	O
cells	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

On	NN	O	O
first	NN	O	O
encounter	NN	O	O
with	NN	O	O
antigen	NN	O	O
,	NN	O	O
naive	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
(	NN	O	I-cell_type
Th	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
differentiate	NN	O	O
into	NN	O	O
cytokine-producing	NN	O	B-cell_line
effector	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Two	NN	O	O
types	NN	O	O
of	NN	O	O
effector	NN	O	O
cells	NN	O	O
characterized	NN	O	O
by	NN	O	O
their	NN	O	O
distinct	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
profiles	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

Th1	NN	O	O
cells	NN	O	O
produce	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
Th2	NN	O	B-cell_line
cells	NN	O	I-cell_line
produce	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-5	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-13	NN	O	B-protein
.	NN	O	O

In	NN	O	O
many	NN	O	O
pathological	NN	O	O
situations	NN	O	O
,	NN	O	O
the	NN	O	O
balance	NN	O	O
between	NN	O	O
Th1	NN	O	O
and	NN	O	O
Th2	NN	O	O
immune	NN	O	O
responses	NN	O	O
determines	NN	O	O
the	NN	O	O
outcome	NN	O	O
of	NN	O	O
diverse	NN	O	O
immunologically	NN	O	O
mediated	NN	O	O
clinical	NN	O	O
syndromes	NN	O	O
including	NN	O	O
infectious	NN	O	O
,	NN	O	O
autoimmune	NN	O	O
,	NN	O	O
and	NN	O	O
allergic	NN	O	O
diseases	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
Th1/Th2-like	NN	O	B-protein
cytokines	NN	O	I-protein
has	NN	O	O
remained	NN	O	O
elusive	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
we	NN	O	O
evaluate	NN	O	O
the	NN	O	O
possible	NN	O	O
in	NN	O	O
vivo	NN	O	O
role	NN	O	O
of	NN	O	O
different	NN	O	O
transcription	NN	O	O
factors	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tcf-1	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
differential	NN	O	O
role	NN	O	O
for	NN	O	O
glycogen	NN	O	B-protein
synthase	NN	O	I-protein
kinase-3	NN	O	I-protein
in	NN	O	O
fibroblasts	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Beta-catenin	NN	O	O
is	NN	O	O
the	NN	O	O
vertebrate	NN	O	O
homolog	NN	O	O
of	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-DNA
segment	NN	O	I-DNA
polarity	NN	O	I-DNA
gene	NN	O	I-DNA
Armadillo	NN	O	B-DNA
and	NN	O	O
plays	NN	O	O
roles	NN	O	O
in	NN	O	O
both	NN	O	O
cell-cell	NN	O	O
adhesion	NN	O	O
and	NN	O	O
transduction	NN	O	O
of	NN	O	O
the	NN	O	O
Wnt	NN	O	O
signaling	NN	O	O
cascade	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Lef/Tcf	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
have	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
protein	NN	O	O
partners	NN	O	O
of	NN	O	O
beta-catenin	NN	O	B-protein
,	NN	O	O
explaining	NN	O	O
how	NN	O	O
beta-catenin	NN	O	B-protein
alters	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
Tcf-1	NN	O	B-protein
also	NN	O	O
becomes	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
through	NN	O	O
interaction	NN	O	O
with	NN	O	O
beta-catenin	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
Wnt	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
is	NN	O	O
operational	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
although	NN	O	O
Wnt	NN	O	O
signals	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
negative	NN	O	B-protein
regulatory	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
glycogen	NN	O	B-protein
synthase	NN	O	I-protein
kinase-3beta	NN	O	I-protein
(	NN	O	O
GSK-3beta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
beta-catenin	NN	O	B-protein
,	NN	O	O
we	NN	O	O
find	NN	O	O
no	NN	O	O
evidence	NN	O	O
for	NN	O	O
involvement	NN	O	O
of	NN	O	O
GSK-3beta	NN	O	B-protein
in	NN	O	O
Tcf-mediated	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

That	NN	O	O
is	NN	O	O
,	NN	O	O
a	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
GSK-3beta	NN	O	I-protein
does	NN	O	O
not	NN	O	O
specifically	NN	O	O
activate	NN	O	O
Tcf	NN	O	O
transcription	NN	O	O
and	NN	O	O
stimuli	NN	O	O
(	NN	O	O
lithium	NN	O	O
or	NN	O	O
phytohemagglutinin	NN	O	B-protein
)	NN	O	O
that	NN	O	O
inhibit	NN	O	O
GSK-3beta	NN	O	B-protein
activity	NN	O	O
also	NN	O	O
do	NN	O	O
not	NN	O	O
activate	NN	O	O
Tcf	NN	O	B-DNA
reporter	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
GSK-3beta	NN	O	B-protein
is	NN	O	O
insufficient	NN	O	O
to	NN	O	O
activate	NN	O	O
Tcf-dependent	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
in	NN	O	O
C57MG	NN	O	B-cell_line
fibroblast	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
lithium	NN	O	O
inactivates	NN	O	O
GSK-3beta	NN	O	B-protein
and	NN	O	O
induces	NN	O	O
Tcf-controlled	NN	O	O
transcription	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
demonstration	NN	O	O
that	NN	O	O
lithium	NN	O	O
can	NN	O	O
alter	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
Tcf-responsive	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
points	NN	O	O
to	NN	O	O
a	NN	O	O
difference	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
Wnt	NN	O	O
signaling	NN	O	O
between	NN	O	O
fibroblasts	NN	O	B-cell_type
and	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

SHP2-interacting	NN	O	B-protein
transmembrane	NN	O	I-protein
adaptor	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
SIT	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
novel	NN	O	O
disulfide-linked	NN	O	B-protein
dimer	NN	O	I-protein
regulating	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
express	NN	O	O
several	NN	O	O
low	NN	O	B-protein
molecular	NN	O	I-protein
weight	NN	O	I-protein
transmembrane	NN	O	I-protein
adaptor	NN	O	I-protein
proteins	NN	O	I-protein
that	NN	O	O
recruit	NN	O	O
src	NN	O	B-protein
homology	NN	O	I-protein
(	NN	O	I-protein
SH	NN	O	I-protein
)	NN	O	I-protein
2	NN	O	I-protein
domain-containing	NN	O	I-protein
intracellular	NN	O	I-protein
molecules	NN	O	I-protein
to	NN	O	O
the	NN	O	O
cell	NN	O	O
membrane	NN	O	O
via	NN	O	O
tyrosine-based	NN	O	O
signaling	NN	O	O
motifs	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
here	NN	O	O
a	NN	O	O
novel	NN	O	O
molecule	NN	O	O
of	NN	O	O
this	NN	O	O
group	NN	O	O
termed	NN	O	O
SIT	NN	O	B-protein
(	NN	O	O
SHP2	NN	O	B-protein
interacting	NN	O	I-protein
transmembrane	NN	O	I-protein
adaptor	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
.	NN	O	O

SIT	NN	O	B-protein
is	NN	O	O
a	NN	O	O
disulfide-linked	NN	O	B-protein
homodimeric	NN	O	I-protein
glycoprotein	NN	O	I-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

After	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
by	NN	O	O
src	NN	O	B-protein
and	NN	O	O
possibly	NN	O	O
syk	NN	O	B-protein
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
SIT	NN	O	B-protein
recruits	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
domain-containing	NN	O	I-protein
tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
SHP2	NN	O	B-protein
via	NN	O	O
an	NN	O	O
immunoreceptor	NN	O	O
tyrosine-based	NN	O	O
inhibition	NN	O	O
motif	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
SIT	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
downmodulates	NN	O	O
T	NN	O	O
cell	NN	O	O
receptor-	NN	O	O
and	NN	O	O
phytohemagglutinin-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
signaling	NN	O	O
processes	NN	O	O
that	NN	O	O
are	NN	O	O
probably	NN	O	O
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
phospholipase	NN	O	B-protein
C	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
binding	NN	O	O
of	NN	O	O
SHP2	NN	O	B-protein
to	NN	O	O
SIT	NN	O	B-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
induction	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
SIT	NN	O	B-protein
not	NN	O	O
only	NN	O	O
regulates	NN	O	O
NF-AT	NN	O	B-protein
activity	NN	O	O
but	NN	O	O
also	NN	O	O
controls	NN	O	O
NF-AT	NN	O	B-protein
unrelated	NN	O	O
pathways	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
involving	NN	O	O
SHP2	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Extracellular-regulated	NN	O	B-protein
kinase	NN	O	I-protein
1/2	NN	O	I-protein
,	NN	O	O
Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
c-Jun	NN	O	B-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
IL-6	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	O
monocytes	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
possible	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	O
extracellular-regulated	NN	O	B-protein
kinase	NN	O	I-protein
1/2	NN	O	I-protein
(	NN	O	O
ERK1/2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
c-	NN	O	O
Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
in	NN	O	O
mediating	NN	O	O
IL-6	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
in	NN	O	O
particular	NN	O	O
their	NN	O	O
role	NN	O	O
in	NN	O	O
enhancing	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

Freshly	NN	O	O
isolated	NN	O	O
monocytes	NN	O	O
treated	NN	O	O
with	NN	O	O
the	NN	O	O
protein	NN	O	O
phosphatase	NN	O	O
inhibitor	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
secreted	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
coincided	NN	O	O
with	NN	O	O
enhanced	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
with	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
ERK1/2	NN	O	B-protein
and	NN	O	I-protein
JNK	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
ERK	NN	O	O
pathway-specific	NN	O	O
inhibitor	NN	O	O
PD98059	NN	O	O
inhibited	NN	O	O
IL-6	NN	O	B-protein
secretion	NN	O	O
from	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Transient	NN	O	O
overexpression	NN	O	O
of	NN	O	O
inactive	NN	O	O
mutants	NN	O	O
of	NN	O	O
either	NN	O	O
Raf-1	NN	O	B-protein
or	NN	O	O
JNK1	NN	O	B-protein
showed	NN	O	O
that	NN	O	O
both	NN	O	O
pathways	NN	O	O
were	NN	O	O
involved	NN	O	O
in	NN	O	O
kappa	NN	O	O
B-dependent	NN	O	O
IL-6	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
PD98059	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
Raf1/MEK1/	NN	O	O
ERK1/2	NN	O	B-protein
pathway	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
but	NN	O	O
,	NN	O	O
rather	NN	O	O
,	NN	O	O
acted	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
shown	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-mediated	NN	O	O
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
both	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
promoter	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
on	NN	O	O
its	NN	O	O
own	NN	O	O
,	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
both	NN	O	O
serine	NN	O	O
kinase	NN	O	O
activity	NN	O	O
and	NN	O	O
interaction	NN	O	O
with	NN	O	O
c-Jun	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
okadaic	NN	O	O
acid-induced	NN	O	O
IL-6	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
at	NN	O	O
least	NN	O	O
partly	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
ERK1/2	NN	O	B-protein
and	NN	O	O
JNK	NN	O	B-protein
pathway-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcriptional	NN	O	O
capacity	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
JNK	NN	O	B-protein
pathway	NN	O	O
may	NN	O	O
regulate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-mediated	NN	O	O
gene	NN	O	O
transcription	NN	O	O
through	NN	O	O
its	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	O
EBNA2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
protein	NN	O	O
required	NN	O	O
for	NN	O	O
B	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
immortalization	NN	O	O
,	NN	O	O
induces	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
interferon	NN	O	I-protein
in	NN	O	O
Burkitt	NN	O	B-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	O
EBNA2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
protein	NN	O	O
involved	NN	O	O
in	NN	O	O
cell	NN	O	O
transformation	NN	O	O
,	NN	O	O
interferes	NN	O	O
with	NN	O	O
the	NN	O	O
cellular	NN	O	O
response	NN	O	O
to	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
interferons	NN	O	I-protein
(	NN	O	O
IFN-alpha/beta	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
conditionally	NN	O	O
expressed	NN	O	O
EBNA2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
IFN	NN	O	B-protein
response	NN	O	O
in	NN	O	O
Burkitt	NN	O	B-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
EBNA2	NN	O	B-protein
led	NN	O	O
to	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
endogenous	NN	O	O
or	NN	O	O
transfected	NN	O	O
IFN-stimulated	NN	O	B-DNA
genes	NN	O	I-DNA
(	NN	O	O
ISGs	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
genes	NN	O	O
which	NN	O	O
contain	NN	O	O
within	NN	O	O
their	NN	O	O
promoters	NN	O	B-DNA
either	NN	O	O
the	NN	O	O
interferon-stimulated	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
ISRE	NN	O	B-DNA
)	NN	O	O
or	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
interferon	NN	O	I-DNA
activation	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
GAS	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

In	NN	O	O
search	NN	O	O
of	NN	O	O
a	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
ISGs	NN	O	B-DNA
,	NN	O	O
we	NN	O	O
observed	NN	O	O
an	NN	O	O
EBNA2	NN	O	B-protein
-dependent	NN	O	O
synthesis	NN	O	O
of	NN	O	O
IFN-beta	NN	O	O
mRNA	NN	O	O
at	NN	O	O
low	NN	O	O
levels	NN	O	O
and	NN	O	O
the	NN	O	O
secretion	NN	O	O
of	NN	O	O
low	NN	O	O
amounts	NN	O	O
of	NN	O	O
IFN	NN	O	O
.	NN	O	O

A	NN	O	O
transfected	NN	O	O
IFN-beta	NN	O	O
promoter	NN	O	O
responded	NN	O	O
to	NN	O	O
EBNA2	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
sequence	NN	O	O
closely	NN	O	O
resembling	NN	O	O
a	NN	O	O
RBP-Jkappa	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
pinpointed	NN	O	O
as	NN	O	O
a	NN	O	O
potential	NN	O	O
target	NN	O	O
of	NN	O	O
EBNA2	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

EBNA2	NN	O	B-protein
-dependent	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-beta	NN	O	B-DNA
promoter	NN	O	I-DNA
occurred	NN	O	O
in	NN	O	O
EBV-negative	NN	O	B-cell_line
Burkitt	NN	O	I-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
other	NN	O	O
EBV	NN	O	B-DNA
genes	NN	O	I-DNA
were	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IFN-beta	NN	O	B-protein
synthesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

PGG-glucan	NN	O	O
,	NN	O	O
a	NN	O	O
soluble	NN	O	O
beta-	NN	O	O
(	NN	O	O
1	NN	O	O
,	NN	O	O
3	NN	O	O
)	NN	O	O
-glucan	NN	O	O
,	NN	O	O
enhances	NN	O	O
the	NN	O	O
oxidative	NN	O	O
burst	NN	O	O
response	NN	O	O
,	NN	O	O
microbicidal	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
activates	NN	O	O
an	NN	O	O
NF-kappa	NN	O	B-protein
B-like	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
PMN	NN	O	I-cell_type
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
glycosphingolipid	NN	O	B-protein
beta-	NN	O	I-protein
(	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	I-protein
3	NN	O	I-protein
)	NN	O	I-protein
-glucan	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

PGG-Glucan	NN	O	O
,	NN	O	O
a	NN	O	O
soluble	NN	O	O
beta-	NN	O	O
(	NN	O	O
1	NN	O	O
,	NN	O	O
6	NN	O	O
)	NN	O	O
-branched	NN	O	O
beta-	NN	O	O
(	NN	O	O
1	NN	O	O
,	NN	O	O
3	NN	O	O
)	NN	O	O
-linked	NN	O	O
glucose	NN	O	O
homopolymer	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
cell	NN	O	O
wall	NN	O	O
of	NN	O	O
the	NN	O	O
yeast	NN	O	O
Saccharomyces	NN	O	O
cerevisiae	NN	O	O
,	NN	O	O
is	NN	O	O
an	NN	O	O
immunomodulator	NN	O	O
which	NN	O	O
enhances	NN	O	O
leukocyte	NN	O	O
anti-infective	NN	O	O
activity	NN	O	O
and	NN	O	O
enhances	NN	O	O
myeloid	NN	O	O
and	NN	O	O
megakaryocyte	NN	O	O
progenitor	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
human	NN	O	O
whole	NN	O	O
blood	NN	O	O
with	NN	O	O
PGG-Glucan	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
the	NN	O	O
oxidative	NN	O	O
burst	NN	O	O
response	NN	O	O
of	NN	O	O
subsequently	NN	O	B-cell_type
isolated	NN	O	I-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
to	NN	O	O
both	NN	O	O
soluble	NN	O	O
and	NN	O	O
particulate	NN	O	O
activators	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
and	NN	O	O
increased	NN	O	O
leukocyte	NN	O	O
microbicidal	NN	O	O
activity	NN	O	O
.	NN	O	O

No	NN	O	O
evidence	NN	O	O
for	NN	O	O
inflammatory	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
was	NN	O	O
obtained	NN	O	O
under	NN	O	O
these	NN	O	O
conditions	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
PGG-Glucan	NN	O	O
induced	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
an	NN	O	O
NF-kappaB-like	NN	O	B-protein
nuclear	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
purified	NN	O	B-cell_type
human	NN	O	I-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
3H-PGG-Glucan	NN	O	O
to	NN	O	O
human	NN	O	O
leukocyte	NN	O	O
membranes	NN	O	O
was	NN	O	O
specific	NN	O	O
,	NN	O	O
concentration-dependent	NN	O	O
,	NN	O	O
saturable	NN	O	O
,	NN	O	O
and	NN	O	O
high	NN	O	O
affinity	NN	O	O
(	NN	O	O
Kd	NN	O	O
approximately	NN	O	O
6	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
specific	NN	O	O
to	NN	O	O
the	NN	O	O
glycosphingolipid	NN	O	O
lactosylceramide	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
inhibit	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB-like	NN	O	B-protein
factor	NN	O	I-protein
by	NN	O	O
PGG-Glucan	NN	O	O
,	NN	O	O
and	NN	O	O
ligand	NN	O	O
binding	NN	O	O
data	NN	O	O
,	NN	O	O
including	NN	O	O
polysaccharide	NN	O	O
specificity	NN	O	O
,	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
PGG-Glucan	NN	O	O
binding	NN	O	O
moiety	NN	O	O
was	NN	O	O
lactosylceramide	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
PGG-Glucan	NN	O	O
enhances	NN	O	O
neutrophil	NN	O	O
anti-microbial	NN	O	O
functions	NN	O	O
and	NN	O	O
that	NN	O	O
interaction	NN	O	O
between	NN	O	O
this	NN	O	O
beta-glucan	NN	O	O
and	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
glycosphingolipid	NN	O	O
lactosylceramide	NN	O	O
present	NN	O	O
at	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoids	NN	O	O
promote	NN	O	O
nonphlogistic	NN	O	O
phagocytosis	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

Phagocyte	NN	O	O
recognition	NN	O	O
,	NN	O	O
uptake	NN	O	O
,	NN	O	O
and	NN	O	O
nonphlogistic	NN	O	O
degradation	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
other	NN	O	O
leukocytes	NN	O	B-cell_type
undergoing	NN	O	O
apoptosis	NN	O	O
promote	NN	O	O
the	NN	O	O
resolution	NN	O	O
of	NN	O	O
inflammation	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
assessed	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
anti-inflammatory	NN	O	O
glucocorticoids	NN	O	O
on	NN	O	O
this	NN	O	O
leukocyte	NN	O	O
clearance	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
``	NN	O	O
semimature	NN	O	O
''	NN	O	O
5-day	NN	O	O
human	NN	O	B-cell_type
monocyte-derived	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
(	NN	O	O
M	NN	O	O
phi	NN	O	O
)	NN	O	O
for	NN	O	O
24	NN	O	O
h	NN	O	O
with	NN	O	O
methylprednisolone	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
and	NN	O	O
hydrocortisone	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
nonglucocorticoid	NN	O	O
steroids	NN	O	O
aldosterone	NN	O	O
,	NN	O	O
estradiol	NN	O	O
,	NN	O	O
and	NN	O	O
progesterone	NN	O	O
,	NN	O	O
potentiated	NN	O	O
phagocytosis	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
effects	NN	O	O
were	NN	O	O
specific	NN	O	O
in	NN	O	O
that	NN	O	O
the	NN	O	O
potentiated	NN	O	O
phagocytosis	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
was	NN	O	O
completely	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
antagonist	NN	O	O
RU38486	NN	O	O
,	NN	O	O
and	NN	O	O
glucocorticoids	NN	O	O
did	NN	O	O
not	NN	O	O
promote	NN	O	O
5-day	NN	O	O
M	NN	O	O
phi	NN	O	O
ingestion	NN	O	O
of	NN	O	O
opsonized	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
.	NN	O	O

Similar	NN	O	O
glucocorticoid-mediated	NN	O	O
potentiation	NN	O	O
was	NN	O	O
observed	NN	O	O
with	NN	O	O
5-day	NN	O	O
M	NN	O	O
phi	NN	O	O
uptake	NN	O	O
of	NN	O	O
alternative	NN	O	O
apoptotic	NN	O	O
``	NN	O	O
targets	NN	O	O
''	NN	O	O
(	NN	O	O
eosinophils	NN	O	B-cell_type
and	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
and	NN	O	O
in	NN	O	O
uptake	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
by	NN	O	O
alternative	NN	O	O
phagocytes	NN	O	B-cell_type
(	NN	O	O
human	NN	O	B-cell_type
glomerular	NN	O	I-cell_type
mesangial	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
murine	NN	O	B-cell_line
M	NN	O	I-cell_line
phi	NN	O	I-cell_line
elicited	NN	O	O
into	NN	O	O
the	NN	O	O
peritoneum	NN	O	O
or	NN	O	O
derived	NN	O	O
from	NN	O	O
bone	NN	O	O
marrow	NN	O	O
)	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
methylprednisolone-mediated	NN	O	O
enhancement	NN	O	O
of	NN	O	O
the	NN	O	O
uptake	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
did	NN	O	O
not	NN	O	O
trigger	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
the	NN	O	O
chemokines	NN	O	B-protein
IL-8	NN	O	B-protein
and	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein-1	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
longer-term	NN	O	O
potentiation	NN	O	O
by	NN	O	O
methylprednisolone	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
maturing	NN	O	B-cell_line
human	NN	O	I-cell_line
monocyte-derived	NN	O	I-cell_line
M	NN	O	I-cell_line
phi	NN	O	I-cell_line
,	NN	O	O
with	NN	O	O
greater	NN	O	O
increases	NN	O	O
in	NN	O	O
5-day	NN	O	O
M	NN	O	B-cell_line
phi	NN	O	I-cell_line
uptake	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-cell_type
cells	NN	O	I-cell_type
being	NN	O	O
observed	NN	O	O
the	NN	O	O
earlier	NN	O	O
glucocorticoids	NN	O	O
were	NN	O	O
added	NN	O	O
during	NN	O	O
monocyte	NN	O	O
maturation	NN	O	O
into	NN	O	O
M	NN	O	B-cell_line
phi	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
potentiation	NN	O	O
of	NN	O	O
nonphlogistic	NN	O	O
clearance	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
by	NN	O	O
phagocytes	NN	O	B-cell_type
is	NN	O	O
a	NN	O	O
hitherto	NN	O	O
unrecognized	NN	O	O
property	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
that	NN	O	O
has	NN	O	O
potential	NN	O	O
implications	NN	O	O
for	NN	O	O
therapies	NN	O	O
aimed	NN	O	O
at	NN	O	O
promoting	NN	O	O
the	NN	O	O
resolution	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

1	NN	O	O
,	NN	O	O
25-Dihydroxyvitamin	NN	O	O
D3	NN	O	O
induces	NN	O	O
differentiation	NN	O	O
of	NN	O	O
a	NN	O	O
retinoic	NN	O	B-cell_line
acid-resistant	NN	O	I-cell_line
acute	NN	O	I-cell_line
promyelocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
UF-1	NN	O	B-cell_line
)	NN	O	O
associated	NN	O	O
with	NN	O	O
expression	NN	O	O
of	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
WAF1/CIP1	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
p27	NN	O	B-protein
(	NN	O	I-protein
KIP1	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
resistance	NN	O	O
is	NN	O	O
a	NN	O	O
serious	NN	O	O
problem	NN	O	O
for	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
who	NN	O	O
are	NN	O	O
receiving	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
and	NN	O	O
strategies	NN	O	O
to	NN	O	O
overcome	NN	O	O
RA	NN	O	O
resistance	NN	O	O
by	NN	O	O
APL	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
still	NN	O	O
unclear	NN	O	O
.	NN	O	O

The	NN	O	O
biologic	NN	O	O
effects	NN	O	O
of	NN	O	O
RA	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
two	NN	O	O
distinct	NN	O	O
families	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
factors	NN	O	I-protein
:	NN	O	O
RA	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
RARs	NN	O	B-protein
)	NN	O	O
and	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RXRs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

RXRs	NN	O	B-protein
heterodimerize	NN	O	O
with	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25-dihydroxyvitamin	NN	O	I-protein
D3	NN	O	I-protein
[	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
]	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
enabling	NN	O	O
their	NN	O	O
efficient	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
cyclin-dependent	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
cdk	NN	O	I-protein
)	NN	O	I-protein
inhibitor	NN	O	I-protein
p21	NN	O	B-protein
(	NN	O	I-protein
WAF1/CIP1	NN	O	I-protein
)	NN	O	I-protein
has	NN	O	O
a	NN	O	O
vitamin	NN	O	B-DNA
D3-responsive	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
VDRE	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
its	NN	O	O
promoter	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
enhances	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
WAF1/CIP1	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
induces	NN	O	O
differentiation	NN	O	O
of	NN	O	O
selected	NN	O	O
myeloid	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
established	NN	O	O
a	NN	O	O
novel	NN	O	O
APL	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
UF-1	NN	O	B-cell_line
)	NN	O	O
with	NN	O	O
features	NN	O	O
of	NN	O	O
RA	NN	O	O
resistance	NN	O	O
.	NN	O	O

1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
can	NN	O	O
induce	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
and	NN	O	O
G1	NN	O	O
arrest	NN	O	O
of	NN	O	O
UF-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
differentiation	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
toward	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

This	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3-induced	NN	O	O
G1	NN	O	O
arrest	NN	O	O
is	NN	O	O
enhanced	NN	O	O
by	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
.	NN	O	O

Also	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
mol/L	NN	O	O
)	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
RA	NN	O	O
markedly	NN	O	O
inhibits	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
in	NN	O	O
a	NN	O	O
dose-	NN	O	O
and	NN	O	O
time-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Associated	NN	O	O
with	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
WAF1/CIP1	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
p27	NN	O	B-RNA
(	NN	O	I-RNA
KIP1	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
increased	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
p21	NN	O	B-RNA
(	NN	O	I-RNA
WAF1/CIP1	NN	O	I-RNA
)	NN	O	I-RNA
and	NN	O	I-RNA
p27	NN	O	I-RNA
(	NN	O	I-RNA
KIP1	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
increased	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
WAF1/CIP1	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
p27	NN	O	B-protein
(	NN	O	I-protein
KIP1	NN	O	I-protein
)	NN	O	I-protein
transcripts	NN	O	O
were	NN	O	O
induced	NN	O	O
after	NN	O	O
6	NN	O	O
hours	NN	O	O
'	NN	O	O
exposure	NN	O	O
to	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
and	NN	O	O
then	NN	O	O
decreased	NN	O	O
to	NN	O	O
basal	NN	O	O
levels	NN	O	O
over	NN	O	O
48	NN	O	O
hours	NN	O	O
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
experiments	NN	O	O
showed	NN	O	O
that	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
WAF1/CIP1	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	O
levels	NN	O	O
increased	NN	O	O
and	NN	O	O
became	NN	O	O
detectable	NN	O	O
after	NN	O	O
12	NN	O	O
hours	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
treatment	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
p27	NN	O	B-protein
(	NN	O	I-protein
KIP1	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	O
was	NN	O	O
much	NN	O	O
more	NN	O	O
gradual	NN	O	O
and	NN	O	O
sustained	NN	O	O
in	NN	O	O
UF-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
and	NN	O	O
RA	NN	O	O
markedly	NN	O	O
enhanced	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
p27	NN	O	B-protein
(	NN	O	I-protein
KIP1	NN	O	I-protein
)	NN	O	I-protein
transcript	NN	O	O
and	NN	O	O
protein	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
levels	NN	O	O
induced	NN	O	O
by	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
alone	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
exogenous	NN	O	O
p27	NN	O	B-protein
(	NN	O	I-protein
KIP1	NN	O	I-protein
)	NN	O	I-protein
expression	NN	O	O
can	NN	O	O
enhance	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
CD11b	NN	O	B-protein
antigen	NN	O	I-protein
in	NN	O	O
myeloid	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
RA	NN	O	O
alone	NN	O	O
can	NN	O	O
induce	NN	O	O
G1	NN	O	O
arrest	NN	O	O
of	NN	O	O
UF-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
;	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
did	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
an	NN	O	O
increase	NN	O	O
of	NN	O	O
p21	NN	O	B-protein
(	NN	O	I-protein
WAF1/CIP1	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
p27	NN	O	B-protein
(	NN	O	I-protein
KIP1	NN	O	I-protein
)	NN	O	I-protein
transcript	NN	O	O
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
in	NN	O	O
RA-resistant	NN	O	O
cells	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
induces	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
cdk	NN	O	B-protein
inhibitors	NN	O	I-protein
,	NN	O	O
which	NN	O	O
mediates	NN	O	O
a	NN	O	O
G1	NN	O	O
arrest	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
differentiation	NN	O	O
of	NN	O	O
RA-resistant	NN	O	O
UF-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
toward	NN	O	O
mature	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
neutrophil-endothelial	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
induced	NN	O	O
by	NN	O	O
redox	NN	O	O
imbalance	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
implicated	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
intracellular	NN	O	O
thiols	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
and	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
endothelial	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

This	NN	O	O
study	NN	O	O
was	NN	O	O
designed	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
changes	NN	O	O
in	NN	O	O
endothelial	NN	O	O
cell	NN	O	O
glutathione	NN	O	O
(	NN	O	O
GSH	NN	O	O
)	NN	O	O
or	NN	O	O
oxidized	NN	O	O
glutathione	NN	O	O
(	NN	O	O
GSSG	NN	O	O
)	NN	O	O
can	NN	O	O
alter	NN	O	O
neutrophil	NN	O	O
adhesivity	NN	O	O
and	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
that	NN	O	O
underlies	NN	O	O
this	NN	O	O
GSSG/GSH-induced	NN	O	O
adhesion	NN	O	O
response	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
umbilical	NN	O	I-cell_line
vein	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cell	NN	O	I-cell_line
(	NN	O	I-cell_line
HUVEC	NN	O	I-cell_line
)	NN	O	I-cell_line
monolayers	NN	O	I-cell_line
for	NN	O	O
6	NN	O	O
hours	NN	O	O
with	NN	O	O
0.2	NN	O	O
mmol/L	NN	O	O
diamide	NN	O	O
and	NN	O	O
1	NN	O	O
mmol/L	NN	O	O
buthionine	NN	O	O
sulfoximine	NN	O	O
(	NN	O	O
BSO	NN	O	O
)	NN	O	O
decreased	NN	O	O
GSH	NN	O	O
levels	NN	O	O
and	NN	O	O
increased	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
GSSG	NN	O	O
to	NN	O	O
GSH	NN	O	O
without	NN	O	O
cell	NN	O	O
toxicity	NN	O	O
.	NN	O	O

These	NN	O	O
redox	NN	O	O
changes	NN	O	O
are	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
observed	NN	O	O
with	NN	O	O
anoxia/reoxygenation	NN	O	O
.	NN	O	O

Diamide	NN	O	O
plus	NN	O	O
BSO-induced	NN	O	O
thiol/disulfide	NN	O	O
imbalance	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
biphasic	NN	O	O
increase	NN	O	O
in	NN	O	O
neutrophil	NN	O	B-cell_type
adhesion	NN	O	O
to	NN	O	O
HUVECs	NN	O	B-cell_line
with	NN	O	O
peak	NN	O	O
responses	NN	O	O
observed	NN	O	O
at	NN	O	O
15	NN	O	O
minutes	NN	O	O
(	NN	O	O
phase	NN	O	O
1	NN	O	O
)	NN	O	O
and	NN	O	O
240	NN	O	O
minutes	NN	O	O
(	NN	O	O
phase	NN	O	O
2	NN	O	O
)	NN	O	O
.	NN	O	O

N-Acetylcysteine	NN	O	O
treatment	NN	O	O
attenuated	NN	O	O
neutrophil	NN	O	B-cell_type
adhesion	NN	O	O
in	NN	O	O
both	NN	O	O
phases	NN	O	O
,	NN	O	O
which	NN	O	O
indicated	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
GSH	NN	O	O
in	NN	O	O
the	NN	O	O
adhesion	NN	O	O
responses	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
phase	NN	O	O
1	NN	O	O
adhesion	NN	O	O
was	NN	O	O
inversely	NN	O	O
correlated	NN	O	O
with	NN	O	O
GSH	NN	O	O
levels	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
the	NN	O	O
GSSG/GSH	NN	O	O
ratio	NN	O	O
,	NN	O	O
whereas	NN	O	O
phase	NN	O	O
2	NN	O	O
neutrophil	NN	O	B-cell_type
adhesion	NN	O	O
was	NN	O	O
positively	NN	O	O
correlated	NN	O	O
with	NN	O	O
GSSG/GSH	NN	O	O
ratio	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
GSH	NN	O	O
levels	NN	O	O
.	NN	O	O

Intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
and	NN	O	O
P-selectin-specific	NN	O	B-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
attenuated	NN	O	O
the	NN	O	O
increased	NN	O	O
neutrophil	NN	O	B-cell_type
adhesion	NN	O	O
during	NN	O	O
both	NN	O	O
phases	NN	O	O
,	NN	O	O
whereas	NN	O	O
an	NN	O	O
anti-E-selectin	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
also	NN	O	O
attenuated	NN	O	O
the	NN	O	O
phase	NN	O	O
2	NN	O	O
response	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
with	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
and	NN	O	O
cycloheximide	NN	O	O
or	NN	O	O
with	NN	O	O
competing	NN	O	O
ds-oligonucleotides	NN	O	O
that	NN	O	O
contained	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
or	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
cognate	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
significantly	NN	O	O
attenuated	NN	O	O
the	NN	O	O
phase	NN	O	O
2	NN	O	O
response	NN	O	O
,	NN	O	O
which	NN	O	O
implicated	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Surface	NN	O	O
expression	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
,	NN	O	O
P-selectin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
E-selectin	NN	O	B-protein
on	NN	O	O
HUVECs	NN	O	B-cell_line
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
phase	NN	O	O
1	NN	O	O
and	NN	O	O
2	NN	O	O
neutrophil	NN	O	B-cell_type
adhesion	NN	O	O
responses	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
changes	NN	O	O
in	NN	O	O
endothelial	NN	O	O
cell	NN	O	O
GSSG/GSH	NN	O	O
cause	NN	O	O
transcription-independent	NN	O	O
and	NN	O	O
transcription-dependent	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
different	NN	O	O
endothelial	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
which	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
2-phase	NN	O	O
neutrophil	NN	O	B-cell_type
-endothelial	NN	O	O
adhesion	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	O
hormone	NN	O	O
suppression	NN	O	O
of	NN	O	O
human	NN	O	O
neutrophil	NN	O	B-cell_type
-mediated	NN	O	O
tumor	NN	O	O
cell	NN	O	O
cytostasis	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
hormones	NN	O	O
on	NN	O	O
neutrophil	NN	O	B-cell_type
-mediated	NN	O	O
tumor	NN	O	O
cell	NN	O	O
cytostasis	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
hydrocortisone	NN	O	O
and	NN	O	O
a	NN	O	O
synthetic	NN	O	O
hormone	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
Dex	NN	O	O
)	NN	O	O
,	NN	O	O
inhibited	NN	O	O
cytostasis	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
Dex	NN	O	O
was	NN	O	O
completely	NN	O	O
reversed	NN	O	O
by	NN	O	O
a	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
antagonist	NN	O	O
,	NN	O	O
RU38486	NN	O	O
.	NN	O	O

To	NN	O	O
clarify	NN	O	O
the	NN	O	O
underlying	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
effects	NN	O	O
of	NN	O	O
Dex	NN	O	O
on	NN	O	O
the	NN	O	O
binding	NN	O	O
avidity	NN	O	O
of	NN	O	O
beta2	NN	O	B-protein
integrin	NN	O	I-protein
on	NN	O	O
the	NN	O	O
neutrophil	NN	O	B-cell_type
surface	NN	O	O
and	NN	O	O
how	NN	O	O
these	NN	O	O
might	NN	O	O
in	NN	O	O
turn	NN	O	O
affect	NN	O	O
neutrophil	NN	O	B-cell_type
-to-tumor	NN	O	O
cell	NN	O	O
binding	NN	O	O
.	NN	O	O

Dex	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
inhibit	NN	O	O
these	NN	O	O
neutrophil	NN	O	B-cell_type
properties	NN	O	O
,	NN	O	O
and	NN	O	O
RU38486	NN	O	O
completely	NN	O	O
suppressed	NN	O	O
both	NN	O	O
forms	NN	O	O
of	NN	O	O
Dex	NN	O	O
inhibition	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
glucocorticoid	NN	O	O
hormone	NN	O	O
inhibition	NN	O	O
of	NN	O	O
neutrophil	NN	O	B-cell_type
-mediated	NN	O	O
tumor	NN	O	O
cell	NN	O	O
cytostasis	NN	O	O
is	NN	O	O
at	NN	O	O
least	NN	O	O
partially	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
lowering	NN	O	O
of	NN	O	O
the	NN	O	O
ligand	NN	O	O
binding	NN	O	O
avidity	NN	O	O
of	NN	O	O
beta2	NN	O	B-protein
integrin	NN	O	I-protein
on	NN	O	O
the	NN	O	O
neutrophil	NN	O	B-cell_type
surface	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
an	NN	O	O
epithelial	NN	O	O
signal	NN	O	O
transducer	NN	O	O
and	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	O
(	NN	O	O
STAT	NN	O	B-protein
)	NN	O	O
pathway	NN	O	O
in	NN	O	O
asthma	NN	O	O
.	NN	O	O

Cytokine	NN	O	B-protein
effects	NN	O	O
on	NN	O	O
immunity	NN	O	O
and	NN	O	O
inflammation	NN	O	O
often	NN	O	O
depend	NN	O	O
on	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
termed	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	O
STATs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
so	NN	O	O
STAT	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
are	NN	O	O
candidates	NN	O	O
for	NN	O	O
influencing	NN	O	O
inflammatory	NN	O	O
disease	NN	O	O
.	NN	O	O

We	NN	O	O
reasoned	NN	O	O
that	NN	O	O
selective	NN	O	O
IFN	NN	O	B-protein
responsiveness	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
STAT	NN	O	B-protein
family	NN	O	I-protein
member	NN	O	I-protein
(	NN	O	O
Stat1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
Stat1-dependent	NN	O	B-DNA
immune-response	NN	O	I-DNA
genes	NN	O	I-DNA
such	NN	O	O
as	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
IFN	NN	O	B-protein
regulatory	NN	O	I-protein
factor-1	NN	O	I-protein
(	NN	O	O
IRF-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
Stat1	NN	O	B-protein
itself	NN	O	O
in	NN	O	O
airway	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
provides	NN	O	O
a	NN	O	O
basis	NN	O	O
for	NN	O	O
detecting	NN	O	O
cytokine	NN	O	B-protein
signaling	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
airway	NN	O	O
disease	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
and	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
epithelial	NN	O	B-protein
Stat1	NN	O	I-protein
(	NN	O	O
but	NN	O	O
not	NN	O	O
other	NN	O	O
control	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
)	NN	O	O
was	NN	O	O
invariably	NN	O	O
activated	NN	O	O
in	NN	O	O
asthmatic	NN	O	O
compared	NN	O	O
with	NN	O	O
normal	NN	O	O
control	NN	O	O
or	NN	O	O
chronic	NN	O	O
bronchitis	NN	O	O
subjects	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
epithelial	NN	O	O
levels	NN	O	O
of	NN	O	O
activated	NN	O	O
Stat1	NN	O	B-protein
correlated	NN	O	O
with	NN	O	O
levels	NN	O	O
of	NN	O	O
expression	NN	O	O
for	NN	O	O
epithelial	NN	O	B-protein
ICAM-1	NN	O	I-protein
,	NN	O	O
IRF-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Stat1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
ICAM-1	NN	O	B-protein
levels	NN	O	O
correlated	NN	O	O
with	NN	O	O
T-cell	NN	O	O
accumulation	NN	O	O
in	NN	O	O
tissue	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
only	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
or	NN	O	O
IFN-gamma-producing	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
detected	NN	O	O
in	NN	O	O
airway	NN	O	O
tissue	NN	O	O
in	NN	O	O
all	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
therefore	NN	O	O
provide	NN	O	O
initial	NN	O	O
evidence	NN	O	O
linking	NN	O	O
abnormal	NN	O	O
behavior	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
pathways	NN	O	O
for	NN	O	O
cytokine	NN	O	B-protein
signaling	NN	O	O
to	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
an	NN	O	O
inflammatory	NN	O	O
disease	NN	O	O
.	NN	O	O

In	NN	O	O
that	NN	O	O
context	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
also	NN	O	O
change	NN	O	O
the	NN	O	O
current	NN	O	O
scheme	NN	O	O
for	NN	O	O
asthma	NN	O	O
pathogenesis	NN	O	O
to	NN	O	O
one	NN	O	O
that	NN	O	O
must	NN	O	O
include	NN	O	O
a	NN	O	O
localized	NN	O	O
gain	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
signal	NN	O	O
ordinarily	NN	O	O
used	NN	O	O
for	NN	O	O
a	NN	O	O
T	NN	O	B-protein
helper	NN	O	I-protein
1-type	NN	O	I-protein
cytokine	NN	O	I-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
allergy-driven	NN	O	O
overproduction	NN	O	O
of	NN	O	O
T	NN	O	B-protein
helper	NN	O	I-protein
2-type	NN	O	I-protein
cytokines	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Leukocyte	NN	O	B-cell_type
populations	NN	O	I-cell_type
,	NN	O	O
hormone	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
eutopic	NN	O	O
and	NN	O	O
ectopic	NN	O	O
first	NN	O	O
trimester	NN	O	O
human	NN	O	O
pregnancies	NN	O	O
.	NN	O	O

The	NN	O	O
implantation	NN	O	O
of	NN	O	O
trophoblast	NN	O	B-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
extrauterine	NN	O	O
sites	NN	O	O
still	NN	O	O
results	NN	O	O
in	NN	O	O
decidualization	NN	O	O
.	NN	O	O

The	NN	O	O
objective	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
compare	NN	O	O
decidualization	NN	O	O
at	NN	O	O
eutopic	NN	O	O
and	NN	O	O
ectopic	NN	O	O
implantation	NN	O	O
sites	NN	O	O
.	NN	O	O

Tissues	NN	O	O
from	NN	O	O
women	NN	O	O
undergoing	NN	O	O
elective	NN	O	O
termination	NN	O	O
of	NN	O	O
uterine	NN	O	O
pregnancy	NN	O	O
and	NN	O	O
from	NN	O	O
women	NN	O	O
with	NN	O	O
ectopic	NN	O	O
pregnancy	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
detect	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cells	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
maintenance	NN	O	O
of	NN	O	O
pregnancy	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
BCL-2+	NN	O	B-cell_type
,	NN	O	I-cell_type
CD56+	NN	O	I-cell_type
,	NN	O	I-cell_type
CD3+	NN	O	I-cell_type
,	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
and	NN	O	I-cell_type
CD68+	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
oestrogen	NN	O	B-protein
(	NN	O	I-protein
ER	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
progesterone	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
PR	NN	O	B-protein
)	NN	O	O
by	NN	O	O
immunohistochemistry	NN	O	O
.	NN	O	O

In-situ	NN	O	O
detection	NN	O	O
of	NN	O	O
fragmented	NN	O	O
DNA	NN	O	O
was	NN	O	O
performed	NN	O	O
to	NN	O	O
identify	NN	O	O
apoptotic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
percentage	NN	O	O
of	NN	O	O
CD3+	NN	O	B-cell_type
cells	NN	O	I-cell_type
among	NN	O	O
all	NN	O	O
immunocompetent	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
tubal	NN	O	O
epithelium	NN	O	O
was	NN	O	O
46.6	NN	O	O
%	NN	O	O
(	NN	O	O
39.9	NN	O	O
%	NN	O	O
of	NN	O	O
CD3+	NN	O	B-cell_type
were	NN	O	O
also	NN	O	O
CD8+	NN	O	B-cell_type
)	NN	O	O
;	NN	O	O
the	NN	O	O
other	NN	O	O
53.4	NN	O	O
%	NN	O	O
were	NN	O	O
CD68+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CD56+	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
undetectable	NN	O	O
in	NN	O	O
ectopic	NN	O	O
decidua	NN	O	O
at	NN	O	O
the	NN	O	O
feto-maternal	NN	O	O
interface	NN	O	O
in	NN	O	O
ectopic	NN	O	O
tissue	NN	O	O
.	NN	O	O

In	NN	O	O
uterine	NN	O	O
decidua	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
29.9	NN	O	O
%	NN	O	O
CD3+	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
2.2	NN	O	O
%	NN	O	O
of	NN	O	O
CD3+	NN	O	O
were	NN	O	O
CD8+	NN	O	O
)	NN	O	O
,	NN	O	O
51.6	NN	O	O
%	NN	O	O
CD56+	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
18.5	NN	O	O
%	NN	O	O
CD68+	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
ratio	NN	O	O
of	NN	O	O
BCL2+	NN	O	B-cell_type
to	NN	O	O
CD3+	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
ectopic	NN	O	O
pregnancy	NN	O	O
was	NN	O	O
0.41	NN	O	O
.	NN	O	O

In	NN	O	O
uterine	NN	O	O
pregnancy	NN	O	O
,	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
BCL-2	NN	O	B-protein
to	NN	O	O
CD3	NN	O	O
was	NN	O	O
0.44	NN	O	O
and	NN	O	O
0.39	NN	O	O
for	NN	O	O
CD56	NN	O	O
.	NN	O	O

Tissues	NN	O	O
from	NN	O	O
both	NN	O	O
ectopic	NN	O	O
and	NN	O	O
uterine	NN	O	O
pregnancies	NN	O	O
were	NN	O	O
positive	NN	O	O
for	NN	O	O
PR	NN	O	B-protein
.	NN	O	O

Fewer	NN	O	O
apoptotic	NN	O	B-cell_type
cell	NN	O	I-cell_type
bodies	NN	O	I-cell_type
were	NN	O	O
present	NN	O	O
in	NN	O	O
ectopic	NN	O	O
pregnancy	NN	O	O
.	NN	O	O

The	NN	O	O
use	NN	O	O
of	NN	O	O
tissue	NN	O	O
obtained	NN	O	O
from	NN	O	O
ectopic	NN	O	O
pregnancy	NN	O	O
may	NN	O	O
become	NN	O	O
an	NN	O	O
excellent	NN	O	O
model	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
trophoblast	NN	O	O
invasion	NN	O	O
in	NN	O	O
eutopic	NN	O	O
pregnancies	NN	O	O
.	NN	O	O

-DOCSTART-	O

Angiotensin	NN	O	B-protein
II	NN	O	I-protein
activates	NN	O	O
the	NN	O	O
proinflammatory	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
renin	NN	O	B-protein
-angiotensin	NN	O	O
system	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
atherosclerosis	NN	O	O
.	NN	O	O

A	NN	O	O
common	NN	O	O
feature	NN	O	O
of	NN	O	O
all	NN	O	O
stages	NN	O	O
of	NN	O	O
atherosclerosis	NN	O	O
is	NN	O	O
inflammation	NN	O	O
of	NN	O	O
the	NN	O	O
vessel	NN	O	O
wall	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	O
factor	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
participates	NN	O	O
in	NN	O	O
most	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
involved	NN	O	O
in	NN	O	O
inflammation	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
therefore	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
angiotensin	NN	O	B-protein
(	NN	O	I-protein
ANG	NN	O	I-protein
)	NN	O	I-protein
II	NN	O	I-protein
on	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
major	NN	O	O
cellular	NN	O	O
component	NN	O	O
of	NN	O	O
human	NN	O	O
atheroma	NN	O	O
,	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

ANG	NN	O	B-protein
II	NN	O	I-protein
,	NN	O	O
like	NN	O	O
TNFalpha	NN	O	B-protein
,	NN	O	O
caused	NN	O	O
rapid	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
by	NN	O	O
Ficoll	NN	O	O
density	NN	O	O
gradient	NN	O	O
.	NN	O	O

This	NN	O	O
ANG	NN	O	B-protein
II	NN	O	I-protein
effect	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
angiotensin	NN	O	O
AT1	NN	O	O
receptor	NN	O	O
antagonist	NN	O	O
losartan	NN	O	O
.	NN	O	O

Specificity	NN	O	O
of	NN	O	O
ANG	NN	O	B-protein
II	NN	O	I-protein
-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
ascertained	NN	O	O
by	NN	O	O
supershift	NN	O	O
and	NN	O	O
competition	NN	O	O
experiments	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
ANG	NN	O	B-protein
II	NN	O	I-protein
stimulated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
same	NN	O	O
preparation	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
the	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
vasoactive	NN	O	O
peptide	NN	O	O
ANG	NN	O	B-protein
II	NN	O	I-protein
to	NN	O	O
activate	NN	O	O
inflammatory	NN	O	O
pathways	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
activated	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
following	NN	O	O
infection	NN	O	O
with	NN	O	O
Chlamydia	NN	O	O
pneumoniae	NN	O	O
.	NN	O	O

Chlamydia	NN	O	O
pneumoniae	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
respiratory	NN	O	O
pathogen	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
its	NN	O	O
presence	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
atherosclerotic	NN	O	O
lesions	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
characterized	NN	O	O
C.	NN	O	O
pneumoniae-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
demonstrated	NN	O	O
an	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
endothelial	NN	O	B-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
followed	NN	O	O
by	NN	O	O
subsequent	NN	O	O
rolling	NN	O	O
,	NN	O	O
adhesion	NN	O	O
,	NN	O	O
and	NN	O	O
transmigration	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
(	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
granulocytes	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

These	NN	O	O
effects	NN	O	O
were	NN	O	O
blocked	NN	O	O
by	NN	O	O
mAbs	NN	O	B-protein
against	NN	O	O
endothelial	NN	O	B-protein
and/or	NN	O	I-protein
leukocyte	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
(	NN	O	O
beta1	NN	O	B-protein
and	NN	O	I-protein
beta2	NN	O	I-protein
integrins	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
different	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
in	NN	O	O
C.	NN	O	B-cell_type
pneumoniae-infected	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
shown	NN	O	O
:	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
phosphorylated	NN	O	O
p42/p44	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
activation/translocation	NN	O	O
occurred	NN	O	O
within	NN	O	O
10-15	NN	O	O
min	NN	O	O
.	NN	O	O

Increased	NN	O	O
mRNA	NN	O	B-RNA
and	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
VCAM-1	NN	O	B-protein
were	NN	O	O
noted	NN	O	O
within	NN	O	O
hours	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
C.	NN	O	O
pneumoniae	NN	O	O
triggers	NN	O	O
a	NN	O	O
cascade	NN	O	O
of	NN	O	O
events	NN	O	O
that	NN	O	O
could	NN	O	O
lead	NN	O	O
to	NN	O	O
endothelial	NN	O	O
activation	NN	O	O
,	NN	O	O
inflammation	NN	O	O
,	NN	O	O
and	NN	O	O
thrombosis	NN	O	O
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
may	NN	O	O
result	NN	O	O
in	NN	O	O
or	NN	O	O
may	NN	O	O
promote	NN	O	O
atherosclerosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transient	NN	O	O
pseudo-hypoaldosteronism	NN	O	O
following	NN	O	O
resection	NN	O	O
of	NN	O	O
the	NN	O	O
ileum	NN	O	O
:	NN	O	O
normal	NN	O	O
level	NN	O	O
of	NN	O	O
lymphocytic	NN	O	B-protein
aldosterone	NN	O	I-protein
receptors	NN	O	I-protein
outside	NN	O	O
the	NN	O	O
acute	NN	O	O
phase	NN	O	O
.	NN	O	O

Pseudo-hypoaldosteronism	NN	O	O
(	NN	O	O
PHA	NN	O	O
)	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
mineralocorticoid	NN	O	O
resistance	NN	O	O
and	NN	O	O
manifests	NN	O	O
as	NN	O	O
hyponatremia	NN	O	O
and	NN	O	O
hyperkalemia	NN	O	O
with	NN	O	O
increased	NN	O	O
plasma	NN	O	O
aldosterone	NN	O	O
levels	NN	O	O
.	NN	O	O

It	NN	O	O
may	NN	O	O
be	NN	O	O
familial	NN	O	O
or	NN	O	O
secondary	NN	O	O
to	NN	O	O
abnormal	NN	O	O
renal	NN	O	O
sodium	NN	O	O
handling	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
a	NN	O	O
54-year-old	NN	O	O
woman	NN	O	O
with	NN	O	O
multifocal	NN	O	O
cancer	NN	O	O
of	NN	O	O
the	NN	O	O
colon	NN	O	O
,	NN	O	O
who	NN	O	O
developed	NN	O	O
PHA	NN	O	O
after	NN	O	O
subtotal	NN	O	O
colectomy	NN	O	O
,	NN	O	O
ileal	NN	O	O
resection	NN	O	O
and	NN	O	O
jejunostomy	NN	O	O
.	NN	O	O

She	NN	O	O
was	NN	O	O
treated	NN	O	O
with	NN	O	O
6	NN	O	O
g	NN	O	O
of	NN	O	O
salt	NN	O	O
daily	NN	O	O
to	NN	O	O
prevent	NN	O	O
dehydration	NN	O	O
,	NN	O	O
which	NN	O	O
she	NN	O	O
stopped	NN	O	O
herself	NN	O	O
because	NN	O	O
of	NN	O	O
reduced	NN	O	O
fecal	NN	O	O
losses	NN	O	O
.	NN	O	O

One	NN	O	O
month	NN	O	O
later	NN	O	O
she	NN	O	O
was	NN	O	O
admitted	NN	O	O
with	NN	O	O
signs	NN	O	O
of	NN	O	O
acute	NN	O	O
adrenal	NN	O	O
failure	NN	O	O
,	NN	O	O
i.e.	NN	O	O
fatigue	NN	O	O
,	NN	O	O
severe	NN	O	O
nausea	NN	O	O
,	NN	O	O
blood	NN	O	O
pressure	NN	O	O
of	NN	O	O
80/60	NN	O	O
mmHg	NN	O	O
,	NN	O	O
extracellular	NN	O	O
dehydration	NN	O	O
,	NN	O	O
hyponatremia	NN	O	O
(	NN	O	O
118	NN	O	O
mmol/l	NN	O	O
)	NN	O	O
;	NN	O	O
hyperkalemia	NN	O	O
(	NN	O	O
7.6	NN	O	O
mmol/l	NN	O	O
)	NN	O	O
,	NN	O	O
increased	NN	O	O
blood	NN	O	O
urea	NN	O	O
nitrogen	NN	O	O
(	NN	O	O
BUN	NN	O	O
)	NN	O	O
(	NN	O	O
200	NN	O	O
mg/dl	NN	O	O
)	NN	O	O
and	NN	O	O
creatininemia	NN	O	O
(	NN	O	O
2.5	NN	O	O
mg/dl	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
decreased	NN	O	O
plasma	NN	O	O
bicarbonates	NN	O	O
level	NN	O	O
(	NN	O	O
HCO3-	NN	O	O
:	NN	O	O
16	NN	O	O
mmol/l	NN	O	O
;	NN	O	O
N	NN	O	O
:	NN	O	O
27-30	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
was	NN	O	O
high	NN	O	O
(	NN	O	O
66	NN	O	O
microg/100	NN	O	O
ml	NN	O	O
at	NN	O	O
10	NN	O	O
:	NN	O	O
00	NN	O	O
h	NN	O	O
;	NN	O	O
N	NN	O	O
:	NN	O	O
8-15	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
ACTH	NN	O	O
was	NN	O	O
normal	NN	O	O
(	NN	O	O
13	NN	O	O
pg/ml	NN	O	O
,	NN	O	O
N	NN	O	O
:	NN	O	O
10-60	NN	O	O
)	NN	O	O
;	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
marked	NN	O	O
increase	NN	O	O
in	NN	O	O
plasma	NN	O	O
renin	NN	O	O
activity	NN	O	O
(	NN	O	O
>	NN	O	O
37	NN	O	O
ng/ml/h	NN	O	O
;	NN	O	O
N	NN	O	O
supine	NN	O	O
<	NN	O	O
3	NN	O	O
)	NN	O	O
,	NN	O	O
active	NN	O	O
renin	NN	O	O
(	NN	O	O
869	NN	O	O
pg/ml	NN	O	O
;	NN	O	O
N	NN	O	O
supine	NN	O	O
:	NN	O	O
1.120	NN	O	O
)	NN	O	O
,	NN	O	O
aldosterone	NN	O	O
(	NN	O	O
>	NN	O	O
2000	NN	O	O
pg/ml	NN	O	O
;	NN	O	O
N	NN	O	O
supine	NN	O	O
<	NN	O	O
150	NN	O	O
)	NN	O	O
and	NN	O	O
plasma	NN	O	O
AVP	NN	O	O
(	NN	O	O
20	NN	O	O
pmol/l	NN	O	O
;	NN	O	O
N	NN	O	O
:	NN	O	O
0.5-2.5	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
plasma	NN	O	O
ANH	NN	O	O
level	NN	O	O
was	NN	O	O
38	NN	O	O
pmol/l	NN	O	O
(	NN	O	O
N	NN	O	O
supine	NN	O	O
:	NN	O	O
5-25	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
urinary	NN	O	O
steroidogram	NN	O	O
resulted	NN	O	O
in	NN	O	O
highly	NN	O	O
elevated	NN	O	O
tetrahydrocortisol	NN	O	O
(	NN	O	O
THF	NN	O	O
:	NN	O	O
13.3	NN	O	O
mg/24h	NN	O	O
;	NN	O	O
N	NN	O	O
:	NN	O	O
1.4+/-0.8	NN	O	O
)	NN	O	O
with	NN	O	O
no	NN	O	O
increase	NN	O	O
in	NN	O	O
tetrahydrocortisone	NN	O	O
(	NN	O	O
THE	NN	O	O
:	NN	O	O
3.16	NN	O	O
mg/24h	NN	O	O
;	NN	O	O
N	NN	O	O
:	NN	O	O
2.7+/-2.0	NN	O	O
)	NN	O	O
excretion	NN	O	O
,	NN	O	O
and	NN	O	O
with	NN	O	O
low	NN	O	O
THE/THF	NN	O	O
(	NN	O	O
0.24	NN	O	O
;	NN	O	O
N	NN	O	O
:	NN	O	O
1.87+/-0.36	NN	O	O
)	NN	O	O
and	NN	O	O
alpha	NN	O	O
THF/THF	NN	O	O
(	NN	O	O
0.35	NN	O	O
;	NN	O	O
N	NN	O	O
:	NN	O	O
0.92+/-0.42	NN	O	O
)	NN	O	O
ratios	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
mineralocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
was	NN	O	O
in	NN	O	O
the	NN	O	O
lower	NN	O	O
normal	NN	O	O
range	NN	O	O
for	NN	O	O
age	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
was	NN	O	O
reduced	NN	O	O
.	NN	O	O

Small-bowel	NN	O	O
resection	NN	O	O
in	NN	O	O
ileostomized	NN	O	O
patients	NN	O	O
causes	NN	O	O
excessive	NN	O	O
fecal	NN	O	O
sodium	NN	O	O
losses	NN	O	O
and	NN	O	O
results	NN	O	O
in	NN	O	O
chronic	NN	O	O
sodium	NN	O	O
depletion	NN	O	O
with	NN	O	O
contraction	NN	O	O
of	NN	O	O
the	NN	O	O
plasma	NN	O	O
volume	NN	O	O
and	NN	O	O
severe	NN	O	O
secondary	NN	O	O
hyperaldosteronism	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
this	NN	O	O
hyperaldosteronism	NN	O	O
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
hyponatremia	NN	O	O
and	NN	O	O
hyperkalemia	NN	O	O
suggesting	NN	O	O
PHA	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
major	NN	O	O
importance	NN	O	O
of	NN	O	O
the	NN	O	O
colon	NN	O	O
for	NN	O	O
the	NN	O	O
absorption	NN	O	O
of	NN	O	O
sodium	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
this	NN	O	O
case	NN	O	O
report	NN	O	O
emphasizes	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
syndrome	NN	O	O
of	NN	O	O
acquired	NN	O	O
PHA	NN	O	O
with	NN	O	O
severe	NN	O	O
hyperkalemia	NN	O	O
after	NN	O	O
resection	NN	O	O
of	NN	O	O
the	NN	O	O
ileum	NN	O	O
and	NN	O	O
colon	NN	O	O
responding	NN	O	O
to	NN	O	O
oral	NN	O	O
salt	NN	O	O
supplementation	NN	O	O
;	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
major	NN	O	O
increase	NN	O	O
in	NN	O	O
AVP	NN	O	O
and	NN	O	O
the	NN	O	O
small	NN	O	O
increase	NN	O	O
in	NN	O	O
ANH	NN	O	O
;	NN	O	O
3	NN	O	O
)	NN	O	O
the	NN	O	O
strong	NN	O	O
increase	NN	O	O
in	NN	O	O
urinary	NN	O	O
THF	NN	O	O
with	NN	O	O
low	NN	O	O
THE/THF	NN	O	O
and	NN	O	O
alpha	NN	O	O
THF/THF	NN	O	O
ratios	NN	O	O
;	NN	O	O
4	NN	O	O
)	NN	O	O
the	NN	O	O
normal	NN	O	O
number	NN	O	O
of	NN	O	O
lymphocytic	NN	O	B-protein
mineralocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
outside	NN	O	O
the	NN	O	O
acute	NN	O	O
episode	NN	O	O
.	NN	O	O

-DOCSTART-	O

Modulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
by	NN	O	O
progesterone-induced	NN	O	B-protein
lymphocyte	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Rat	NN	O	O
spleen	NN	O	O
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
express	NN	O	O
progesterone	NN	O	B-protein
receptors	NN	O	I-protein
whose	NN	O	O
concentration	NN	O	O
is	NN	O	O
increased	NN	O	O
greatly	NN	O	O
during	NN	O	O
the	NN	O	O
early	NN	O	O
phase	NN	O	O
of	NN	O	O
pregnancy	NN	O	O
.	NN	O	O

After	NN	O	O
stimulation	NN	O	O
of	NN	O	O
progesterone	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
receptors	NN	O	O
was	NN	O	O
augmented	NN	O	O
2-3	NN	O	O
times	NN	O	O
.	NN	O	O

When	NN	O	O
cells	NN	O	O
were	NN	O	O
cultured	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
progesterone	NN	O	O
they	NN	O	O
released	NN	O	O
a	NN	O	O
soluble	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
inhibited	NN	O	O
cellular	NN	O	O
immunoreactions	NN	O	O
(	NN	O	O
MLR	NN	O	O
,	NN	O	O
CRC	NN	O	O
)	NN	O	O
and	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
by	NN	O	O
spleen-cell	NN	O	B-cell_line
culture	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
factor	NN	O	O
also	NN	O	O
inhibited	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
anti-DNP	NN	O	B-protein
antibodies	NN	O	I-protein
by	NN	O	O
a	NN	O	O
mouse	NN	O	B-cell_line
hybridoma	NN	O	I-cell_line
and	NN	O	O
diminished	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
cells	NN	O	O
in	NN	O	O
phase	NN	O	O
S	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
asymmetric	NN	O	B-protein
molecules	NN	O	I-protein
produced	NN	O	O
by	NN	O	O
the	NN	O	O
hybridoma	NN	O	B-cell_line
remained	NN	O	O
unaltered	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
soluble	NN	O	B-protein
factors	NN	O	I-protein
released	NN	O	O
by	NN	O	O
rat	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
modulate	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
mother	NN	O	O
and	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
mechanism	NN	O	O
that	NN	O	O
protects	NN	O	O
the	NN	O	O
fetus	NN	O	O
against	NN	O	O
antipaternal	NN	O	B-protein
antibodies	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
regulation	NN	O	O
of	NN	O	O
4E-BP1	NN	O	B-protein
and	NN	O	O
4E-BP2	NN	O	B-protein
,	NN	O	O
two	NN	O	O
repressors	NN	O	O
of	NN	O	O
translation	NN	O	O
initiation	NN	O	O
,	NN	O	O
during	NN	O	O
human	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
.	NN	O	O

Human	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
translation	NN	O	O
rate	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
determinant	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
translation	NN	O	O
initiation	NN	O	O
during	NN	O	O
human	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
using	NN	O	O
the	NN	O	O
HL-60	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
U-937	NN	O	B-cell_line
monoblastic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
translation	NN	O	O
rate	NN	O	O
is	NN	O	O
observed	NN	O	O
when	NN	O	O
the	NN	O	O
cells	NN	O	O
are	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
along	NN	O	O
the	NN	O	O
monocytic/macrophage	NN	O	O
pathway	NN	O	O
or	NN	O	O
along	NN	O	O
the	NN	O	O
granulocytic	NN	O	O
pathway	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
in	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
correlates	NN	O	O
with	NN	O	O
specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
two	NN	O	O
repressors	NN	O	O
of	NN	O	O
translation	NN	O	O
initiation	NN	O	O
,	NN	O	O
4E-BP1	NN	O	B-protein
and	NN	O	O
4E-BP2	NN	O	B-protein
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
and	NN	O	O
U-937	NN	O	B-cell_line
cell	NN	O	O
differentiation	NN	O	O
into	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
by	NN	O	O
IFN-gamma	NN	O	B-protein
or	NN	O	O
PMA	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
dephosphorylation	NN	O	O
and	NN	O	O
consequent	NN	O	O
activation	NN	O	O
of	NN	O	O
4E-BP1	NN	O	B-protein
.	NN	O	O

Dephosphorylation	NN	O	O
of	NN	O	O
4E-BP1	NN	O	B-protein
was	NN	O	O
also	NN	O	O
observed	NN	O	O
when	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
into	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
following	NN	O	O
treatment	NN	O	O
with	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
or	NN	O	O
DMSO	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
treatment	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
or	NN	O	O
DMSO	NN	O	O
,	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
decreases	NN	O	O
4E-BP1	NN	O	B-protein
amount	NN	O	O
without	NN	O	O
affecting	NN	O	O
its	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
strongly	NN	O	O
increases	NN	O	O
4E-BP2	NN	O	B-protein
amount	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
differential	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
translational	NN	O	O
machinery	NN	O	O
during	NN	O	O
human	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
,	NN	O	O
specific	NN	O	O
to	NN	O	O
the	NN	O	O
monocytic/macrophage	NN	O	O
pathway	NN	O	O
or	NN	O	O
to	NN	O	O
the	NN	O	O
granulocytic	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nonimmunoglobulin	NN	O	O
gene	NN	O	O
hypermutation	NN	O	O
in	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Somatic	NN	O	O
hypermutation	NN	O	O
is	NN	O	O
the	NN	O	O
most	NN	O	O
critical	NN	O	O
mechanism	NN	O	O
underlying	NN	O	O
the	NN	O	O
diversification	NN	O	O
of	NN	O	O
Ig	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Although	NN	O	O
mutation	NN	O	O
occurs	NN	O	O
specifically	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
the	NN	O	O
germinal	NN	O	O
center	NN	O	O
reaction	NN	O	O
,	NN	O	O
it	NN	O	O
remains	NN	O	O
a	NN	O	O
matter	NN	O	O
of	NN	O	O
debate	NN	O	O
whether	NN	O	O
the	NN	O	O
mutation	NN	O	O
machinery	NN	O	O
also	NN	O	O
targets	NN	O	O
non-Ig	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
noncoding	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
Bcl6	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
different	NN	O	O
subtypes	NN	O	O
of	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
frequent	NN	O	O
hypermutation	NN	O	O
in	NN	O	O
follicular	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
25	NN	O	O
of	NN	O	O
59	NN	O	O
=	NN	O	O
42	NN	O	O
%	NN	O	O
)	NN	O	O
(	NN	O	O
germinal	NN	O	O
center	NN	O	O
cell	NN	O	O
origin	NN	O	O
)	NN	O	O
and	NN	O	O
mucosa-associated	NN	O	O
lymphoid	NN	O	O
tissue	NN	O	O
(	NN	O	O
MALT	NN	O	O
)	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
19	NN	O	O
of	NN	O	O
45	NN	O	O
=	NN	O	O
42	NN	O	O
%	NN	O	O
)	NN	O	O
(	NN	O	O
postgerminal	NN	O	O
center	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
occasionally	NN	O	O
in	NN	O	O
mantle	NN	O	O
cell	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
1	NN	O	O
of	NN	O	O
21	NN	O	O
=	NN	O	O
4.8	NN	O	O
%	NN	O	O
)	NN	O	O
(	NN	O	O
pregerminal	NN	O	O
center	NN	O	O
)	NN	O	O
.	NN	O	O

Most	NN	O	O
mutations	NN	O	O
were	NN	O	O
outside	NN	O	O
the	NN	O	O
motifs	NN	O	O
potentially	NN	O	O
important	NN	O	O
for	NN	O	O
transcription	NN	O	O
,	NN	O	O
suggesting	NN	O	O
they	NN	O	O
were	NN	O	O
not	NN	O	O
important	NN	O	O
in	NN	O	O
lymphomagenesis	NN	O	O
but	NN	O	O
may	NN	O	O
,	NN	O	O
like	NN	O	O
Ig	NN	O	O
mutation	NN	O	O
,	NN	O	O
represent	NN	O	O
an	NN	O	O
inherent	NN	O	O
feature	NN	O	O
of	NN	O	O
the	NN	O	O
lymphoma	NN	O	B-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
their	NN	O	O
normal	NN	O	O
cell	NN	O	O
counterparts	NN	O	O
microdissected	NN	O	O
from	NN	O	O
a	NN	O	O
reactive	NN	O	O
tonsil	NN	O	O
.	NN	O	O

Bcl6	NN	O	O
mutation	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
13	NN	O	O
of	NN	O	O
24	NN	O	O
(	NN	O	O
54	NN	O	O
%	NN	O	O
)	NN	O	O
clones	NN	O	O
from	NN	O	O
the	NN	O	O
germinal	NN	O	O
centre	NN	O	O
but	NN	O	O
only	NN	O	O
in	NN	O	O
1	NN	O	O
of	NN	O	O
24	NN	O	O
(	NN	O	O
4	NN	O	O
%	NN	O	O
)	NN	O	O
clones	NN	O	O
from	NN	O	O
the	NN	O	O
naive	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
mantle	NN	O	O
zone	NN	O	O
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
,	NN	O	O
distribution	NN	O	O
,	NN	O	O
and	NN	O	O
nature	NN	O	O
of	NN	O	O
these	NN	O	O
mutations	NN	O	O
were	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
resulting	NN	O	O
from	NN	O	O
the	NN	O	O
Ig	NN	O	O
hypermutation	NN	O	O
process	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
show	NN	O	O
unequivocal	NN	O	O
evidence	NN	O	O
of	NN	O	O
non-	NN	O	O
Ig	NN	O	B-DNA
gene	NN	O	I-DNA
hypermutation	NN	O	O
in	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
provide	NN	O	O
fresh	NN	O	O
insights	NN	O	O
into	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
hypermutation	NN	O	O
and	NN	O	O
lymphomagenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Impaired	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
DQ2	NN	O	B-protein
and	NN	O	O
DQ8	NN	O	B-protein
-binding	NN	O	O
HSV	NN	O	O
VP16	NN	O	O
peptide	NN	O	O
to	NN	O	O
a	NN	O	O
DQA1*0501/DQB1*0302	NN	O	B-protein
trans	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
heterodimer	NN	O	I-protein
.	NN	O	O

DQalpha	NN	O	B-protein
and	NN	O	I-protein
DQbeta	NN	O	I-protein
trans	NN	O	I-protein
heterodimeric	NN	O	I-protein
HLA-DQ	NN	O	I-protein
molecules	NN	O	I-protein
form	NN	O	O
in	NN	O	O
individuals	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
DQ2	NN	O	B-protein
and	NN	O	O
DQ8	NN	O	B-protein
specificities	NN	O	O
.	NN	O	O

Unique	NN	O	O
functions	NN	O	O
and	NN	O	O
disease	NN	O	O
associations	NN	O	O
have	NN	O	O
been	NN	O	O
postulated	NN	O	O
for	NN	O	O
such	NN	O	O
trans-dimers	NN	O	B-protein
,	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
different	NN	O	O
from	NN	O	O
cis-encoded	NN	O	B-protein
DQ	NN	O	I-protein
molecules	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
corresponding	NN	O	O
haplotypes	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
trans-dimer	NN	O	O
encoded	NN	O	O
by	NN	O	O
HLA-DQA1*0501/DQB1*0302	NN	O	B-protein
to	NN	O	O
bind	NN	O	O
a	NN	O	O
peptide	NN	O	O
antigen	NN	O	O
which	NN	O	O
interacts	NN	O	O
with	NN	O	O
DQ	NN	O	B-protein
molecules	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
both	NN	O	O
parental	NN	O	O
haplotypes	NN	O	O
.	NN	O	O

Markedly	NN	O	O
impaired	NN	O	O
binding	NN	O	O
was	NN	O	O
observed	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
both	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
different	NN	O	O
anchor	NN	O	O
residues	NN	O	O
and	NN	O	O
with	NN	O	O
changes	NN	O	O
in	NN	O	O
levels	NN	O	O
of	NN	O	O
DQ	NN	O	O
cis-	NN	O	O
dimer	NN	O	B-protein
availability	NN	O	O
for	NN	O	O
peptide	NN	O	O
binding	NN	O	O
interactions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Detection	NN	O	O
of	NN	O	O
intracellular	NN	O	B-protein
phosphorylated	NN	O	I-protein
STAT-1	NN	O	I-protein
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
applied	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
to	NN	O	O
the	NN	O	O
investigation	NN	O	O
of	NN	O	O
interferon-gamma	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
approach	NN	O	O
uses	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
that	NN	O	O
distinguish	NN	O	O
between	NN	O	O
the	NN	O	O
native	NN	O	O
and	NN	O	O
phosphorylated	NN	O	O
forms	NN	O	O
of	NN	O	O
STAT-1	NN	O	B-protein
.	NN	O	O

It	NN	O	O
enables	NN	O	O
rapid	NN	O	O
and	NN	O	O
quantitative	NN	O	O
assessment	NN	O	O
of	NN	O	O
STAT-1	NN	O	B-protein
phosphorylation	NN	O	O
on	NN	O	O
a	NN	O	O
discrete	NN	O	O
cell	NN	O	O
basis	NN	O	O
and	NN	O	O
is	NN	O	O
both	NN	O	O
more	NN	O	O
sensitive	NN	O	O
and	NN	O	O
less	NN	O	O
time	NN	O	O
consuming	NN	O	O
than	NN	O	O
immunoblotting	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
it	NN	O	O
allows	NN	O	O
for	NN	O	O
discrimination	NN	O	O
between	NN	O	O
a	NN	O	O
mixture	NN	O	O
of	NN	O	O
cells	NN	O	O
that	NN	O	O
differ	NN	O	O
in	NN	O	O
their	NN	O	O
response	NN	O	O
to	NN	O	O
interferon-gamma	NN	O	B-protein
.	NN	O	O

This	NN	O	O
approach	NN	O	O
should	NN	O	O
allow	NN	O	O
for	NN	O	O
the	NN	O	O
evaluation	NN	O	O
of	NN	O	O
different	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
using	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
monoclonal	NN	O	O
reagents	NN	O	O
that	NN	O	O
are	NN	O	O
specific	NN	O	O
for	NN	O	O
native	NN	O	O
and	NN	O	O
activation	NN	O	B-protein
modified	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Application	NN	O	O
of	NN	O	O
this	NN	O	O
form	NN	O	O
of	NN	O	O
testing	NN	O	O
should	NN	O	O
prove	NN	O	O
valuable	NN	O	O
in	NN	O	O
screening	NN	O	O
for	NN	O	O
signaling	NN	O	O
defects	NN	O	O
in	NN	O	O
selected	NN	O	O
patients	NN	O	O
with	NN	O	O
recurrent	NN	O	O
infections	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
this	NN	O	O
technique	NN	O	O
should	NN	O	O
permit	NN	O	O
dissection	NN	O	O
of	NN	O	O
a	NN	O	O
full	NN	O	O
range	NN	O	O
of	NN	O	O
cellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
at	NN	O	O
the	NN	O	O
protein	NN	O	O
level	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
by	NN	O	O
lipopolysaccharide	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
is	NN	O	O
prevented	NN	O	O
by	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
cytosolic	NN	O	B-protein
phospholipase	NN	O	I-protein
A2	NN	O	I-protein
.	NN	O	O

In	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
lipopolysaccharide	NN	O	O
induces	NN	O	O
synthesis	NN	O	O
and	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
85-kDa	NN	O	O
cytosolic	NN	O	B-protein
phospholipase	NN	O	I-protein
A2	NN	O	I-protein
.	NN	O	O

This	NN	O	O
enzyme	NN	O	O
releases	NN	O	O
arachidonic	NN	O	O
acid	NN	O	O
and	NN	O	O
lyso-phospholipids	NN	O	O
from	NN	O	O
membranes	NN	O	O
which	NN	O	O
are	NN	O	O
metabolized	NN	O	O
to	NN	O	O
eicosanoids	NN	O	O
and	NN	O	O
platelet-activating-factor	NN	O	O
.	NN	O	O

These	NN	O	O
lipid	NN	O	O
mediators	NN	O	O
increase	NN	O	O
activity	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
indicating	NN	O	O
a	NN	O	O
function	NN	O	O
for	NN	O	O
cytosolic	NN	O	B-protein
phospholipase	NN	O	I-protein
A2	NN	O	I-protein
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
and	NN	O	O
inflammation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
previously	NN	O	O
that	NN	O	O
trifluoromethylketone	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
cytosolic	NN	O	B-protein
phospholipase	NN	O	I-protein
A2	NN	O	I-protein
suppressed	NN	O	O
interleukin-1beta	NN	O	O
protein	NN	O	O
and	NN	O	O
steady-state	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
in	NN	O	O
human	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
subcellular	NN	O	O
mechanisms	NN	O	O
were	NN	O	O
analyzed	NN	O	O
by	NN	O	O
which	NN	O	O
trifluoromethylketones	NN	O	O
interfere	NN	O	O
with	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
they	NN	O	O
reduced	NN	O	O
the	NN	O	O
initial	NN	O	O
interleukin-1beta	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcription	NN	O	O
rate	NN	O	O
through	NN	O	O
prevention	NN	O	O
of	NN	O	O
degradation	NN	O	O
of	NN	O	O
inhibitor-kappaB	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
cytosolic	NN	O	O
activation	NN	O	O
,	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
DNA-binding	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
were	NN	O	O
decreased	NN	O	O
.	NN	O	O

Trifluoromethylketones	NN	O	O
ameliorate	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
this	NN	O	O
therapeutic	NN	O	O
potency	NN	O	O
may	NN	O	O
reside	NN	O	O
in	NN	O	O
retention	NN	O	O
of	NN	O	O
inactive	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytosol	NN	O	B-cell_type
thereby	NN	O	O
abrogating	NN	O	O
interleukin-1beta	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cellular	NN	O	O
disposition	NN	O	O
of	NN	O	O
sulphamethoxazole	NN	O	O
and	NN	O	O
its	NN	O	O
metabolites	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
hypersensitivity	NN	O	O
.	NN	O	O

1.	NN	O	O
Bioactivation	NN	O	O
of	NN	O	O
sulphamethoxazole	NN	O	O
(	NN	O	O
SMX	NN	O	O
)	NN	O	O
to	NN	O	O
chemically-reactive	NN	O	O
metabolites	NN	O	O
and	NN	O	O
subsequent	NN	O	O
protein	NN	O	O
conjugation	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
SMX	NN	O	O
hypersensitivity	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
therefore	NN	O	O
examined	NN	O	O
the	NN	O	O
cellular	NN	O	O
metabolism	NN	O	O
,	NN	O	O
disposition	NN	O	O
and	NN	O	O
conjugation	NN	O	O
of	NN	O	O
SMX	NN	O	O
and	NN	O	O
its	NN	O	O
metabolites	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

2.	NN	O	O
Flow	NN	O	O
cytometry	NN	O	O
revealed	NN	O	O
binding	NN	O	O
of	NN	O	O
N-hydroxy	NN	O	O
(	NN	O	O
SMX-NHOH	NN	O	O
)	NN	O	O
and	NN	O	O
nitroso	NN	O	O
(	NN	O	O
SMX-NO	NN	O	O
)	NN	O	O
metabolites	NN	O	O
of	NN	O	O
SMX	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
SMX	NN	O	O
itself	NN	O	O
,	NN	O	O
to	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
viable	NN	O	O
white	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cellular	NN	O	O
haptenation	NN	O	O
by	NN	O	O
SMX-NO	NN	O	O
was	NN	O	O
reduced	NN	O	O
by	NN	O	O
exogenous	NN	O	O
glutathione	NN	O	O
(	NN	O	O
GSH	NN	O	O
)	NN	O	O
.	NN	O	O

3.	NN	O	O
SMX-NHOH	NN	O	O
and	NN	O	O
SMX-NO	NN	O	O
were	NN	O	O
rapidly	NN	O	O
reduced	NN	O	O
back	NN	O	O
to	NN	O	O
the	NN	O	O
parent	NN	O	O
compound	NN	O	O
by	NN	O	O
cysteine	NN	O	O
(	NN	O	O
CYS	NN	O	O
)	NN	O	O
,	NN	O	O
GSH	NN	O	O
,	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
plasma	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
and	NN	O	O
ubiquitous	NN	O	B-protein
bioinactivation	NN	O	O
mechanism	NN	O	O
.	NN	O	O

4.	NN	O	O
Fluorescence	NN	O	O
HPLC	NN	O	O
showed	NN	O	O
that	NN	O	O
SMX-NHOH	NN	O	O
and	NN	O	O
SMX-NO	NN	O	O
depleted	NN	O	O
CYS	NN	O	O
and	NN	O	O
GSH	NN	O	O
in	NN	O	O
buffer	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
in	NN	O	O
cells	NN	O	O
and	NN	O	O
plasma	NN	O	O
.	NN	O	O

5.	NN	O	O
Neutrophil	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
neutrophil	NN	O	O
function	NN	O	O
were	NN	O	O
induced	NN	O	O
at	NN	O	O
lower	NN	O	O
concentrations	NN	O	O
of	NN	O	O
SMX-NHOH	NN	O	O
and	NN	O	O
SMX-NO	NN	O	O
than	NN	O	O
those	NN	O	O
inducing	NN	O	O
loss	NN	O	O
of	NN	O	O
membrane	NN	O	O
viability	NN	O	O
,	NN	O	O
with	NN	O	O
SMX	NN	O	O
having	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
were	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
more	NN	O	O
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
direct	NN	O	O
cytotoxic	NN	O	O
effects	NN	O	O
of	NN	O	O
SMX-NO	NN	O	O
than	NN	O	O
neutrophils	NN	O	B-cell_type
.	NN	O	O

6.	NN	O	O
Partitioning	NN	O	O
of	NN	O	O
SMX-NHOH	NN	O	O
into	NN	O	O
red	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
lower	NN	O	O
than	NN	O	O
with	NN	O	O
the	NN	O	O
hydroxylamine	NN	O	O
of	NN	O	O
dapsone	NN	O	O
.	NN	O	O

7.	NN	O	O
Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
balance	NN	O	O
between	NN	O	O
oxidation	NN	O	O
of	NN	O	O
SMX	NN	O	O
to	NN	O	O
its	NN	O	O
toxic	NN	O	O
metabolites	NN	O	O
and	NN	O	O
their	NN	O	O
reduction	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
protective	NN	O	O
cellular	NN	O	O
mechanism	NN	O	O
.	NN	O	O

If	NN	O	O
an	NN	O	O
imbalance	NN	O	O
exists	NN	O	O
,	NN	O	O
haptenation	NN	O	O
of	NN	O	O
the	NN	O	O
toxic	NN	O	O
metabolites	NN	O	O
to	NN	O	O
bodily	NN	O	O
proteins	NN	O	O
including	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
viable	NN	O	O
cells	NN	O	O
can	NN	O	O
occur	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
result	NN	O	O
in	NN	O	O
drug	NN	O	O
hypersensitivity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Bcl-6	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
reactive	NN	O	O
follicular	NN	O	O
hyperplasia	NN	O	O
,	NN	O	O
follicular	NN	O	O
lymphoma	NN	O	O
,	NN	O	O
and	NN	O	O
angioimmunoblastic	NN	O	O
T-cell	NN	O	O
lymphoma	NN	O	O
with	NN	O	O
hyperplastic	NN	O	O
germinal	NN	O	O
centers	NN	O	O
:	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
intrafollicular	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
and	NN	O	O
their	NN	O	O
altered	NN	O	O
distribution	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
angioimmunoblastic	NN	O	O
T-cell	NN	O	O
lymphoma	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
The	NN	O	O
Bcl-6	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
,	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
phosphoprotein	NN	O	I-protein
,	NN	O	O
is	NN	O	O
expressed	NN	O	O
independently	NN	O	O
of	NN	O	O
Bcl-6	NN	O	B-protein
gene	NN	O	O
rearrangement	NN	O	O
.	NN	O	O

In	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
Bcl-6	NN	O	B-protein
protein	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
(	NN	O	I-cell_type
GC	NN	O	I-cell_type
)	NN	O	I-cell_type
B-cells	NN	O	I-cell_type
and	NN	O	O
10	NN	O	O
%	NN	O	O
to	NN	O	O
15	NN	O	O
%	NN	O	O
of	NN	O	O
CD3/CD4+	NN	O	B-cell_type
intrafollicular	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Interfollicular	NN	O	O
cells	NN	O	O
are	NN	O	O
negative	NN	O	O
for	NN	O	O
Bcl-6	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
except	NN	O	O
for	NN	O	O
rare	NN	O	O
CD3+/CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
reported	NN	O	O
cases	NN	O	O
of	NN	O	O
angioimmunoblastic	NN	O	O
T-cell	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
AITL	NN	O	O
)	NN	O	O
with	NN	O	O
hyperplastic	NN	O	O
GCs	NN	O	O
(	NN	O	O
AITL/GC	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
observed	NN	O	O
that	NN	O	O
borders	NN	O	O
of	NN	O	O
enlarged	NN	O	O
GCs	NN	O	O
were	NN	O	O
ill	NN	O	O
defined	NN	O	O
,	NN	O	O
with	NN	O	O
features	NN	O	O
suggestive	NN	O	O
of	NN	O	O
an	NN	O	O
outward	NN	O	O
migration	NN	O	O
of	NN	O	O
GC	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
surrounding	NN	O	O
interfollicular	NN	O	O
zones	NN	O	O
.	NN	O	O

This	NN	O	O
prompted	NN	O	O
a	NN	O	O
study	NN	O	O
of	NN	O	O
follicular	NN	O	O
borders	NN	O	O
with	NN	O	O
Bcl-6	NN	O	B-protein
staining	NN	O	O
in	NN	O	O
reactive	NN	O	O
follicular	NN	O	O
hyperplasias	NN	O	O
and	NN	O	O
follicular	NN	O	O
lymphomas	NN	O	O
to	NN	O	O
compare	NN	O	O
with	NN	O	O
AITL/GC	NN	O	O
.	NN	O	O

MATERIALS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
Formalin-fixed	NN	O	O
paraffin	NN	O	O
sections	NN	O	O
were	NN	O	O
used	NN	O	O
for	NN	O	O
immunostaining	NN	O	O
of	NN	O	O
Bcl-6	NN	O	B-protein
.	NN	O	O

Six	NN	O	O
cases	NN	O	O
of	NN	O	O
AITL/GC	NN	O	O
,	NN	O	O
12	NN	O	O
nonspecific	NN	O	O
reactive	NN	O	O
follicular	NN	O	O
hyperplasia	NN	O	O
(	NN	O	O
FH	NN	O	O
)	NN	O	O
,	NN	O	O
7	NN	O	O
HIV	NN	O	O
adenopathy	NN	O	O
,	NN	O	O
10	NN	O	O
follicular	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
FL	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
8	NN	O	O
typical	NN	O	O
AITL	NN	O	O
(	NN	O	O
ie	NN	O	O
,	NN	O	O
AITL	NN	O	O
without	NN	O	O
GC	NN	O	O
)	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

Double	NN	O	O
staining	NN	O	O
for	NN	O	O
Bcl-6/CD20	NN	O	B-protein
,	NN	O	O
Bcl-6/CD3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Bcl-6/CD57	NN	O	B-protein
was	NN	O	O
performed	NN	O	O
in	NN	O	O
selected	NN	O	O
cases	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
In	NN	O	O
FH	NN	O	O
and	NN	O	O
HIV	NN	O	O
adenopathy	NN	O	O
,	NN	O	O
staining	NN	O	O
for	NN	O	O
Bcl-6	NN	O	B-protein
revealed	NN	O	O
densely	NN	O	O
populated	NN	O	O
GCs	NN	O	O
with	NN	O	O
well-defined	NN	O	O
and	NN	O	O
regular	NN	O	O
GC	NN	O	O
borders	NN	O	O
,	NN	O	O
whereas	NN	O	O
Bcl-6+	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
rare	NN	O	O
in	NN	O	O
the	NN	O	O
interfollicular	NN	O	O
areas	NN	O	O
.	NN	O	O

An	NN	O	O
occasional	NN	O	O
GC	NN	O	O
with	NN	O	O
an	NN	O	O
ill-defined	NN	O	O
border	NN	O	O
was	NN	O	O
invariably	NN	O	O
surrounded	NN	O	O
by	NN	O	O
a	NN	O	O
broad	NN	O	O
mantle	NN	O	O
zone	NN	O	O
;	NN	O	O
those	NN	O	O
with	NN	O	O
indistinct	NN	O	O
mantle	NN	O	O
zones	NN	O	O
had	NN	O	O
well-defined	NN	O	O
,	NN	O	O
regular	NN	O	O
borders	NN	O	O
.	NN	O	O

In	NN	O	O
FL	NN	O	O
,	NN	O	O
follicles	NN	O	O
were	NN	O	O
densely	NN	O	O
populated	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
borders	NN	O	O
were	NN	O	O
irregular	NN	O	O
,	NN	O	O
with	NN	O	O
some	NN	O	O
Bcl-6+	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
interfollicular	NN	O	O
zones	NN	O	O
.	NN	O	O

In	NN	O	O
AITL/GC	NN	O	O
,	NN	O	O
GCs	NN	O	O
were	NN	O	O
less	NN	O	O
dense	NN	O	O
,	NN	O	O
GC	NN	O	O
borders	NN	O	O
were	NN	O	O
ill	NN	O	O
defined	NN	O	O
and	NN	O	O
irregular	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
interfollicular	NN	O	B-cell_type
Bcl-6+	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
markedly	NN	O	O
increased	NN	O	O
.	NN	O	O

Double	NN	O	O
staining	NN	O	O
revealed	NN	O	O
that	NN	O	O
these	NN	O	O
interfollicular	NN	O	B-cell_type
Bcl-6+	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
AITL/GC	NN	O	O
were	NN	O	O
Bcl6+/CD3+/CD20-/CD57-	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
CD3+	NN	O	B-cell_type
intrafollicular	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
depleted	NN	O	O
in	NN	O	O
AITL/GC	NN	O	O
,	NN	O	O
whereas	NN	O	O
they	NN	O	O
were	NN	O	O
abundant	NN	O	O
in	NN	O	O
FH	NN	O	O
.	NN	O	O

Intrafollicular	NN	O	O
CD57+	NN	O	B-cell_type
cells	NN	O	I-cell_type
did	NN	O	O
not	NN	O	O
stain	NN	O	O
for	NN	O	O
Bcl-6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
were	NN	O	O
also	NN	O	O
depleted	NN	O	O
in	NN	O	O
AITL/GC	NN	O	O
.	NN	O	O

In	NN	O	O
typical	NN	O	O
AITL	NN	O	O
,	NN	O	O
some	NN	O	O
neoplastic	NN	O	O
cells	NN	O	O
were	NN	O	O
positive	NN	O	O
for	NN	O	O
Bcl-6	NN	O	B-protein
,	NN	O	O
showing	NN	O	O
variable	NN	O	O
degrees	NN	O	O
of	NN	O	O
staining	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
GCs	NN	O	O
of	NN	O	O
AITL/GC	NN	O	O
differed	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
other	NN	O	O
reactive	NN	O	O
follicular	NN	O	O
hyperplasias	NN	O	O
and	NN	O	O
follicular	NN	O	O
lymphomas	NN	O	O
,	NN	O	O
and	NN	O	O
staining	NN	O	O
for	NN	O	O
Bcl-6	NN	O	B-protein
was	NN	O	O
useful	NN	O	O
to	NN	O	O
discern	NN	O	O
them	NN	O	O
.	NN	O	O

(	NN	O	O
2	NN	O	O
)	NN	O	O
Intrafollicular	NN	O	B-cell_type
CD3+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
many	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
also	NN	O	O
positive	NN	O	O
for	NN	O	O
Bcl-6	NN	O	B-protein
,	NN	O	O
were	NN	O	O
markedly	NN	O	O
depleted	NN	O	O
in	NN	O	O
AITL/GC	NN	O	O
,	NN	O	O
with	NN	O	O
increased	NN	O	O
interfollicular	NN	O	O
Bcl-6+/CD3+	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
an	NN	O	O
outward	NN	O	O
migration	NN	O	O
of	NN	O	O
intrafollicular	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
this	NN	O	O
condition	NN	O	O
.	NN	O	O

(	NN	O	O
3	NN	O	O
)	NN	O	O
Interfollicular	NN	O	B-cell_type
Bcl-6+/CD3+	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
AITL/GC	NN	O	O
were	NN	O	O
too	NN	O	O
numerous	NN	O	O
to	NN	O	O
be	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
migration	NN	O	O
alone	NN	O	O
,	NN	O	O
suggesting	NN	O	O
local	NN	O	O
proliferation	NN	O	O
.	NN	O	O

(	NN	O	O
4	NN	O	O
)	NN	O	O
Intrafollicular	NN	O	B-cell_type
CD57+	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
negative	NN	O	O
for	NN	O	O
Bcl-6	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
the	NN	O	O
intrafollicular	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
population	NN	O	I-cell_type
.	NN	O	O

(	NN	O	O
5	NN	O	O
)	NN	O	O
Some	NN	O	O
neoplastic	NN	O	O
cells	NN	O	O
in	NN	O	O
AITL	NN	O	O
stained	NN	O	O
for	NN	O	O
Bcl-6	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
Bcl-6	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
this	NN	O	O
tumor	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
cytomegalovirus	NN	O	O
binding	NN	O	O
to	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
induces	NN	O	O
immunoregulatory	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

To	NN	O	O
continue	NN	O	O
our	NN	O	O
investigation	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	O
events	NN	O	O
that	NN	O	O
occur	NN	O	O
following	NN	O	O
human	NN	O	O
CMV	NN	O	O
(	NN	O	O
HCMV	NN	O	O
)	NN	O	O
infection	NN	O	O
,	NN	O	O
we	NN	O	O
focused	NN	O	O
on	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cellular	NN	O	O
activation	NN	O	O
following	NN	O	O
viral	NN	O	O
binding	NN	O	O
to	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

First	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
viral	NN	O	O
binding	NN	O	O
induced	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
immunoregulatory	NN	O	B-cell_type
genes	NN	O	I-cell_type
(	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
A20	NN	O	B-protein
,	NN	O	O
NF-kappaB-p105/p50	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IkappaBalpha	NN	O	B-protein
)	NN	O	O
in	NN	O	O
unactivated	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
that	NN	O	O
neutralizing	NN	O	B-protein
Abs	NN	O	I-protein
to	NN	O	O
the	NN	O	O
major	NN	O	O
HCMV	NN	O	B-protein
glycoproteins	NN	O	I-protein
,	NN	O	O
gB	NN	O	B-protein
(	NN	O	O
UL55	NN	O	B-protein
)	NN	O	O
and	NN	O	O
gH	NN	O	B-protein
(	NN	O	O
UL75	NN	O	B-protein
)	NN	O	O
,	NN	O	O
inhibited	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
.	NN	O	O

Next	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
these	NN	O	O
viral	NN	O	O
ligands	NN	O	O
directly	NN	O	O
up-regulated	NN	O	O
monocyte	NN	O	O
gene	NN	O	O
expression	NN	O	O
upon	NN	O	O
their	NN	O	O
binding	NN	O	O
to	NN	O	O
their	NN	O	O
appropriate	NN	O	O
cellular	NN	O	O
receptors	NN	O	O
.	NN	O	O

We	NN	O	O
then	NN	O	O
investigated	NN	O	O
if	NN	O	O
HCMV	NN	O	O
binding	NN	O	O
also	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
translation	NN	O	O
and	NN	O	O
secretion	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
HCMV	NN	O	O
binding	NN	O	O
to	NN	O	O
monocytes	NN	O	B-cell_type
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
production	NN	O	O
and	NN	O	O
release	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Because	NN	O	O
these	NN	O	O
induced	NN	O	O
gene	NN	O	O
products	NN	O	O
have	NN	O	O
NF-kappaB	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
their	NN	O	O
promoter	NN	O	B-DNA
regions	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
next	NN	O	O
examined	NN	O	O
whether	NN	O	O
there	NN	O	O
was	NN	O	O
an	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
nuclear	NN	O	O
NF-kappaB	NN	O	B-protein
levels	NN	O	O
.	NN	O	O

These	NN	O	O
experiments	NN	O	O
showed	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
fact	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
was	NN	O	O
translocated	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
following	NN	O	O
viral	NN	O	O
binding	NN	O	O
or	NN	O	O
purified	NN	O	O
viral	NN	O	O
ligand	NN	O	O
binding	NN	O	O
.	NN	O	O

Changes	NN	O	O
in	NN	O	O
IkappaBalpha	NN	O	B-protein
levels	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
changes	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
translocation	NN	O	O
.	NN	O	O

Lastly	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
p38	NN	O	B-protein
kinase	NN	O	O
activity	NN	O	O
played	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
IL-1beta	NN	O	B-protein
production	NN	O	O
and	NN	O	O
that	NN	O	O
it	NN	O	O
was	NN	O	O
rapidly	NN	O	O
up-regulated	NN	O	O
following	NN	O	O
infection	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
support	NN	O	O
our	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
HCMV	NN	O	O
initiates	NN	O	O
a	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
that	NN	O	O
leads	NN	O	O
to	NN	O	O
monocyte	NN	O	O
activation	NN	O	O
and	NN	O	O
pinpoints	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
whereby	NN	O	O
HCMV	NN	O	O
infection	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
can	NN	O	O
result	NN	O	O
in	NN	O	O
profound	NN	O	O
pathogenesis	NN	O	O
,	NN	O	O
especially	NN	O	O
in	NN	O	O
chronic	NN	O	O
inflammatory-type	NN	O	O
conditions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
megakaryocytic	NN	O	B-DNA
glycoprotein	NN	O	I-DNA
IX	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
oncogenic	NN	O	B-protein
Ets	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Fli-1	NN	O	B-protein
.	NN	O	O

Glycoprotein	NN	O	B-protein
(	NN	O	I-protein
GP	NN	O	I-protein
)	NN	O	I-protein
IX	NN	O	I-protein
is	NN	O	O
a	NN	O	O
subunit	NN	O	O
of	NN	O	O
the	NN	O	O
von	NN	O	B-protein
Willebrand	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
GPIb-V-IX	NN	O	B-protein
,	NN	O	O
which	NN	O	O
mediates	NN	O	O
adhesion	NN	O	O
of	NN	O	O
platelets	NN	O	B-cell_type
to	NN	O	O
the	NN	O	O
subendothelium	NN	O	O
of	NN	O	O
damaged	NN	O	O
blood	NN	O	O
vessels	NN	O	O
.	NN	O	O

Previous	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
GPIX	NN	O	B-DNA
promoter	NN	O	I-DNA
identified	NN	O	O
a	NN	O	O
functional	NN	O	O
Ets	NN	O	O
site	NN	O	O
that	NN	O	O
,	NN	O	O
when	NN	O	O
disrupted	NN	O	O
,	NN	O	O
reduced	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
that	NN	O	O
regulated	NN	O	O
GPIX	NN	O	B-DNA
promoter	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
transient	NN	O	O
cotransfection	NN	O	O
of	NN	O	O
several	NN	O	O
GPIX	NN	O	B-DNA
promoter/reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
into	NN	O	O
293T	NN	O	B-cell_line
kidney	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
with	NN	O	O
a	NN	O	O
Fli-1	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
shows	NN	O	O
that	NN	O	O
the	NN	O	O
oncogenic	NN	O	O
protein	NN	O	O
Fli-1	NN	O	B-protein
can	NN	O	O
transactivate	NN	O	O
the	NN	O	O
GPIX	NN	O	B-DNA
promoter	NN	O	I-DNA
when	NN	O	O
an	NN	O	O
intact	NN	O	O
GPIX	NN	O	B-DNA
Ets	NN	O	I-DNA
site	NN	O	I-DNA
is	NN	O	O
present	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
Fli-1	NN	O	B-protein
binding	NN	O	O
of	NN	O	O
the	NN	O	O
GPIX	NN	O	B-DNA
Ets	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
identified	NN	O	O
in	NN	O	O
antibody	NN	O	O
supershift	NN	O	O
experiments	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
derived	NN	O	O
from	NN	O	O
hematopoietic	NN	O	B-cell_type
human	NN	O	I-cell_type
erythroleukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Comparative	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
Fli-1	NN	O	B-protein
was	NN	O	O
also	NN	O	O
able	NN	O	O
to	NN	O	O
transactivate	NN	O	O
the	NN	O	O
GPIbalpha	NN	O	B-DNA
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
the	NN	O	O
GPIIb	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Immunoblot	NN	O	O
analysis	NN	O	O
identified	NN	O	O
Fli-1	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
lysates	NN	O	O
derived	NN	O	O
from	NN	O	O
platelets	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
Fli-1	NN	O	B-protein
was	NN	O	O
identified	NN	O	O
immunohistochemically	NN	O	O
in	NN	O	O
megakaryocytes	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
CD34	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
the	NN	O	O
megakaryocyte	NN	O	B-protein
differentiation	NN	O	I-protein
and	NN	O	I-protein
proliferation	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
thrombopoietin	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
Fli-1	NN	O	B-protein
is	NN	O	O
likely	NN	O	O
to	NN	O	O
regulate	NN	O	O
lineage-specific	NN	O	B-DNA
genes	NN	O	I-DNA
during	NN	O	O
megakaryocytopoiesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

According	NN	O	O
to	NN	O	O
current	NN	O	O
models	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
capacity	NN	O	O
of	NN	O	O
I	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
would	NN	O	O
be	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
retention	NN	O	O
of	NN	O	O
Rel/NF-kappaB	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytosol	NN	O	B-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
I	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
has	NN	O	O
also	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
cell	NN	O	O
lines	NN	O	O
and	NN	O	O
when	NN	O	O
overexpressed	NN	O	O
by	NN	O	O
transient	NN	O	O
transfection	NN	O	O
.	NN	O	O

To	NN	O	O
gain	NN	O	O
better	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
I	NN	O	I-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
a	NN	O	O
physiological	NN	O	O
context	NN	O	O
we	NN	O	O
have	NN	O	O
analysed	NN	O	O
its	NN	O	O
presence	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
PBL	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
I	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
PBL	NN	O	B-cell_type
by	NN	O	O
different	NN	O	O
techniques	NN	O	O
:	NN	O	O
Western	NN	O	O
blot	NN	O	O
,	NN	O	O
indirect	NN	O	O
immunofluorescence	NN	O	O
and	NN	O	O
electron	NN	O	O
microscopy	NN	O	O
.	NN	O	O

Low	NN	O	O
levels	NN	O	O
of	NN	O	O
nuclear	NN	O	O
I	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
in	NN	O	O
resting	NN	O	O
cells	NN	O	O
whereas	NN	O	O
a	NN	O	O
superinduction	NN	O	O
was	NN	O	O
obtained	NN	O	O
after	NN	O	O
PMA	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
nuclear	NN	O	O
pool	NN	O	O
of	NN	O	O
I	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
showed	NN	O	O
a	NN	O	O
higher	NN	O	O
stability	NN	O	O
than	NN	O	O
cytosolic	NN	O	O
I	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
was	NN	O	O
partially	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
resynthesis	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
I	NN	O	I-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
did	NN	O	O
not	NN	O	O
inhibit	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
and	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
was	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IkappaBbeta	NN	O	B-protein
at	NN	O	O
the	NN	O	O
nuclear	NN	O	O
level	NN	O	O
.	NN	O	O

Immunoprecipitation	NN	O	O
experiments	NN	O	O
failed	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
an	NN	O	O
association	NN	O	O
between	NN	O	O
nuclear	NN	O	O
I	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
proteins	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
in	NN	O	O
resting	NN	O	O
and	NN	O	O
PMA-activated	NN	O	B-cell_type
human	NN	O	I-cell_type
PBL	NN	O	I-cell_type
,	NN	O	O
I	NN	O	B-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
)	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
in	NN	O	O
an	NN	O	O
apparently	NN	O	O
inactive	NN	O	O
form	NN	O	O
unable	NN	O	O
to	NN	O	O
disrupt	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
from	NN	O	O
DNA	NN	O	O
.	NN	O	O

-DOCSTART-	O

Impaired	NN	O	O
fetal	NN	O	B-cell_type
thymocyte	NN	O	I-cell_type
development	NN	O	O
after	NN	O	O
efficient	NN	O	O
adenovirus-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

We	NN	O	O
introduce	NN	O	O
a	NN	O	O
new	NN	O	O
experimental	NN	O	O
system	NN	O	O
combining	NN	O	O
adenovirus-mediated	NN	O	O
gene	NN	O	O
transfer	NN	O	O
and	NN	O	O
fetal	NN	O	O
thymic	NN	O	O
organ	NN	O	O
culture	NN	O	O
(	NN	O	O
FTOC	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
system	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
efficiently	NN	O	O
express	NN	O	O
in	NN	O	O
developing	NN	O	O
thymocytes	NN	O	O
a	NN	O	O
mutant	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibitor	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
mut-I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
and	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
maturation	NN	O	O
defects	NN	O	O
occurring	NN	O	O
when	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
is	NN	O	O
inhibited	NN	O	O
during	NN	O	O
fetal	NN	O	O
development	NN	O	O
.	NN	O	O

Fetal	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
infected	NN	O	O
with	NN	O	O
adenovirus	NN	O	O
containing	NN	O	O
mut-I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
were	NN	O	O
found	NN	O	O
to	NN	O	O
develop	NN	O	O
normally	NN	O	O
until	NN	O	O
the	NN	O	O
CD44	NN	O	B-protein
-CD25+	NN	O	O
,	NN	O	O
CD4	NN	O	B-protein
-	NN	O	O
CD8	NN	O	B-protein
-	NN	O	O
double-negative	NN	O	O
stage	NN	O	O
,	NN	O	O
while	NN	O	O
production	NN	O	O
of	NN	O	O
more	NN	O	O
mature	NN	O	O
double-positive	NN	O	O
and	NN	O	O
single-positive	NN	O	O
populations	NN	O	O
was	NN	O	O
strongly	NN	O	O
decreased	NN	O	O
.	NN	O	O

Proliferation	NN	O	O
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
cells	NN	O	O
in	NN	O	O
cycle	NN	O	O
appeared	NN	O	O
normal	NN	O	O
,	NN	O	O
as	NN	O	O
did	NN	O	O
rearrangement	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
beta-chain	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
much	NN	O	O
higher	NN	O	O
in	NN	O	O
FTOC	NN	O	O
infected	NN	O	O
with	NN	O	O
adenovirus	NN	O	O
containing	NN	O	O
mut-I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
than	NN	O	O
in	NN	O	O
FTOC	NN	O	O
infected	NN	O	O
with	NN	O	O
a	NN	O	O
control	NN	O	O
virus	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
ensuring	NN	O	O
the	NN	O	O
differentiation	NN	O	O
and	NN	O	O
survival	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
in	NN	O	O
the	NN	O	O
early	NN	O	O
stages	NN	O	O
of	NN	O	O
their	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
relationship	NN	O	O
between	NN	O	O
Ca2+-ATPase	NN	O	B-protein
and	NN	O	O
freely	NN	O	O
exchangeable	NN	O	O
Ca2+	NN	O	O
in	NN	O	O
the	NN	O	O
dense	NN	O	O
tubules	NN	O	O
:	NN	O	O
a	NN	O	O
study	NN	O	O
in	NN	O	O
platelets	NN	O	B-cell_type
from	NN	O	O
women	NN	O	O
.	NN	O	O

The	NN	O	O
main	NN	O	O
aims	NN	O	O
of	NN	O	O
this	NN	O	O
work	NN	O	O
were	NN	O	O
to	NN	O	O
examine	NN	O	O
in	NN	O	O
women	NN	O	O
:	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
freely	NN	O	O
exchangeable	NN	O	O
Ca2+	NN	O	O
(	NN	O	O
FECa2+	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
dense	NN	O	O
tubules	NN	O	O
and	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
sarco	NN	O	B-protein
(	NN	O	I-protein
endo	NN	O	I-protein
)	NN	O	I-protein
plasmic	NN	O	I-protein
reticulum	NN	O	I-protein
(	NN	O	I-protein
SER	NN	O	I-protein
)	NN	O	I-protein
Ca2+-ATPase	NN	O	I-protein
(	NN	O	I-protein
SERCA	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
platelets	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
relationship	NN	O	O
of	NN	O	O
these	NN	O	O
parameters	NN	O	O
with	NN	O	O
blood	NN	O	O
pressure	NN	O	O
and	NN	O	O
serum	NN	O	B-protein
lipoproteins	NN	O	I-protein
.	NN	O	O

Platelets	NN	O	O
from	NN	O	O
14	NN	O	O
white	NN	O	O
and	NN	O	O
13	NN	O	O
black	NN	O	O
women	NN	O	O
in	NN	O	O
good	NN	O	O
health	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

The	NN	O	O
FECa2+	NN	O	O
was	NN	O	O
measured	NN	O	O
as	NN	O	O
the	NN	O	O
ionomycin-evoked	NN	O	O
Ca2+	NN	O	O
release	NN	O	O
(	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
thapsigargin	NN	O	O
)	NN	O	O
in	NN	O	O
Ca2+-free	NN	O	O
medium	NN	O	O
.	NN	O	O

SERCA	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
measured	NN	O	O
as	NN	O	O
the	NN	O	O
thapsigargin	NN	O	O
sensitive	NN	O	O
,	NN	O	O
Ca2+	NN	O	O
dependent	NN	O	O
and	NN	O	O
ouabain	NN	O	O
resistant	NN	O	O
,	NN	O	O
ATP	NN	O	O
hydrolyses	NN	O	O
in	NN	O	O
platelet	NN	O	O
membranes	NN	O	O
.	NN	O	O

Relative	NN	O	O
expressions	NN	O	O
of	NN	O	O
SERCA	NN	O	B-protein
2	NN	O	O
and	NN	O	O
3	NN	O	O
isoforms	NN	O	O
and	NN	O	O
Ras-related	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
Rap	NN	O	I-protein
)	NN	O	I-protein
1	NN	O	I-protein
in	NN	O	O
platelet	NN	O	O
membranes	NN	O	O
were	NN	O	O
determined	NN	O	O
by	NN	O	O
Western	NN	O	O
immunoblots	NN	O	O
.	NN	O	O

Highly	NN	O	O
significant	NN	O	O
correlations	NN	O	O
were	NN	O	O
observed	NN	O	O
for	NN	O	O
FECa2+	NN	O	O
in	NN	O	O
the	NN	O	O
dense	NN	O	O
tubules	NN	O	O
with	NN	O	O
:	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
maximal	NN	O	O
reaction	NN	O	O
velocity	NN	O	O
(	NN	O	O
Vmax	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
SERCA	NN	O	B-protein
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.592	NN	O	O
,	NN	O	O
P	NN	O	O
=	NN	O	O
.0014	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
2	NN	O	O
)	NN	O	O
Rapl	NN	O	B-protein
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.551	NN	O	O
,	NN	O	O
P	NN	O	O
=	NN	O	O
.0035	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
negative	NN	O	O
correlations	NN	O	O
were	NN	O	O
observed	NN	O	O
between	NN	O	O
FECa2+	NN	O	O
in	NN	O	O
the	NN	O	O
dense	NN	O	O
tubules	NN	O	O
and	NN	O	O
age	NN	O	O
.	NN	O	O

No	NN	O	O
correlations	NN	O	O
were	NN	O	O
observed	NN	O	O
for	NN	O	O
these	NN	O	O
variables	NN	O	O
with	NN	O	O
blood	NN	O	O
pressure	NN	O	O
or	NN	O	O
serum	NN	O	B-protein
lipoproteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
the	NN	O	O
FECa2+	NN	O	O
and	NN	O	O
the	NN	O	O
Vmax	NN	O	O
of	NN	O	O
the	NN	O	O
SERCA	NN	O	B-protein
are	NN	O	O
reliable	NN	O	O
indicators	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
load	NN	O	O
in	NN	O	O
platelets	NN	O	B-cell_type
from	NN	O	O
women	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
women	NN	O	O
,	NN	O	O
unlike	NN	O	O
previous	NN	O	O
observations	NN	O	O
in	NN	O	O
men	NN	O	O
,	NN	O	O
these	NN	O	O
platelet	NN	O	O
parameters	NN	O	O
are	NN	O	O
not	NN	O	O
correlated	NN	O	O
with	NN	O	O
blood	NN	O	O
pressure	NN	O	O
and	NN	O	O
serum	NN	O	B-protein
lipoproteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Cobalt	NN	O	O
chloride-induced	NN	O	O
signaling	NN	O	O
in	NN	O	O
endothelium	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
augmented	NN	O	O
adherence	NN	O	O
of	NN	O	O
sickle	NN	O	B-cell_type
red	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
of	NN	O	O
monocyte-like	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
PAF-receptor	NN	O	O
antagonist	NN	O	O
.	NN	O	O

In	NN	O	O
response	NN	O	O
to	NN	O	O
hypoxia	NN	O	O
,	NN	O	O
sickle	NN	O	B-cell_type
red	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
SS	NN	O	B-cell_type
RBC	NN	O	I-cell_type
)	NN	O	O
and	NN	O	O
leukocytes	NN	O	B-cell_type
exhibit	NN	O	O
increased	NN	O	O
adherence	NN	O	O
to	NN	O	O
the	NN	O	O
vascular	NN	O	O
endothelium	NN	O	O
,	NN	O	O
while	NN	O	O
diapedesis	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
through	NN	O	O
the	NN	O	O
blood	NN	O	O
vessel	NN	O	O
increases	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
cellular	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
caused	NN	O	O
by	NN	O	O
hypoxia	NN	O	O
is	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

We	NN	O	O
utilized	NN	O	O
CoCl2	NN	O	O
as	NN	O	O
a	NN	O	O
mimetic	NN	O	O
molecule	NN	O	O
for	NN	O	O
hypoxia	NN	O	O
to	NN	O	O
study	NN	O	O
cellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HUVEC	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
CoCl2	NN	O	O
at	NN	O	O
2	NN	O	O
mM	NN	O	O
concentration	NN	O	O
induced	NN	O	O
the	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
CAMs	NN	O	B-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
HUVEC	NN	O	B-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
CoCl2	NN	O	O
also	NN	O	O
caused	NN	O	O
time-dependent	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MAP	NN	O	I-protein
)	NN	O	I-protein
kinase	NN	O	I-protein
isoform	NN	O	O
ERK2	NN	O	B-protein
without	NN	O	O
significantly	NN	O	O
affecting	NN	O	O
ERK1	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
ERK2	NN	O	B-protein
is	NN	O	O
the	NN	O	O
preferred	NN	O	O
substrate	NN	O	O
for	NN	O	O
upstream	NN	O	B-protein
kinase	NN	O	I-protein
of	NN	O	O
the	NN	O	O
MAPK	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

Inhibitors	NN	O	O
of	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
PD98059	NN	O	O
)	NN	O	O
or	NN	O	O
platelet-activating	NN	O	O
factor	NN	O	O
(	NN	O	O
PAF	NN	O	O
)	NN	O	O
-	NN	O	O
receptor	NN	O	O
antagonist	NN	O	O
(	NN	O	O
CV3988	NN	O	O
)	NN	O	O
inhibited	NN	O	O
the	NN	O	O
CoCl2-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Augmented	NN	O	O
expression	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
led	NN	O	O
to	NN	O	O
increased	NN	O	O
SS	NN	O	B-cell_type
RBC	NN	O	I-cell_type
adhesion	NN	O	O
,	NN	O	O
inhibitable	NN	O	O
by	NN	O	O
a	NN	O	O
VCAM-1	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
CoCl2	NN	O	O
caused	NN	O	O
a	NN	O	O
two-	NN	O	O
to	NN	O	O
threefold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
of	NN	O	O
monocyte-like	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
a	NN	O	O
twentyfold	NN	O	O
increase	NN	O	O
in	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
platelet	NN	O	B-protein
endothelial	NN	O	I-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
(	NN	O	O
PECAM-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
an	NN	O	O
antibody	NN	O	O
to	NN	O	O
PECAM-1	NN	O	B-protein
.	NN	O	O

Both	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
PECAM-1	NN	O	B-protein
and	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
CoCl2	NN	O	O
were	NN	O	O
inhibited	NN	O	O
by	NN	O	O
protein	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
GF109203X	NN	O	O
)	NN	O	O
and	NN	O	O
augmented	NN	O	O
by	NN	O	O
protein	NN	O	O
phosphatase	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
Calyculin	NN	O	O
A	NN	O	O
)	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggests	NN	O	O
that	NN	O	O
CoCl2-induced	NN	O	O
cellular	NN	O	O
signals	NN	O	O
directing	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
in	NN	O	O
HUVEC	NN	O	B-cell_type
involve	NN	O	O
downstream	NN	O	O
activation	NN	O	O
of	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
while	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
PECAM-1	NN	O	B-protein
occurs	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
PAF-receptor	NN	O	O
antagonist	NN	O	O
inhibits	NN	O	O
the	NN	O	O
CoCl2	NN	O	O
-or	NN	O	O
hypoxia-induced	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
adhesion	NN	O	O
of	NN	O	O
SS	NN	O	B-cell_type
RBC	NN	O	I-cell_type
,	NN	O	O
PECAM-1	NN	O	B-protein
phosphorylation	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
concomitant	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Bacterial	NN	O	O
lipopolysaccharide	NN	O	O
activates	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
through	NN	O	O
interleukin-1	NN	O	B-protein
signaling	NN	O	I-protein
mediators	NN	O	I-protein
in	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
dermal	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
.	NN	O	O

Bacterial	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-mediated	NN	O	O
immune	NN	O	O
responses	NN	O	O
,	NN	O	O
including	NN	O	O
activation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
Gram-negative	NN	O	O
bacteria-induced	NN	O	O
sepsis	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
cytokine	NN	O	O
release	NN	O	O
from	NN	O	O
LPS-responsive	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
critical	NN	O	O
step	NN	O	O
for	NN	O	O
endotoxic	NN	O	O
effects	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
and	NN	O	O
involvement	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
LPS	NN	O	O
signaling	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
dermal	NN	O	I-cell_type
microvessel	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HDMEC	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
THP-1	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

LPS	NN	O	O
stimulation	NN	O	O
of	NN	O	O
HDMEC	NN	O	B-cell_type
and	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
initiated	NN	O	O
an	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	I-protein
-like	NN	O	O
NF-kappaB	NN	O	B-protein
signaling	NN	O	O
cascade	NN	O	O
.	NN	O	O

In	NN	O	O
transient	NN	O	O
cotransfection	NN	O	O
experiments	NN	O	O
,	NN	O	O
dominant	NN	O	O
negative	NN	O	O
mutants	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
,	NN	O	O
including	NN	O	O
MyD88	NN	O	B-protein
,	NN	O	O
IRAK	NN	O	B-protein
,	NN	O	O
IRAK2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TRAF6	NN	O	B-protein
inhibited	NN	O	O
both	NN	O	O
IL-1	NN	O	B-protein
-and	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
-luciferase	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
not	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
mutant	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
that	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
TNF	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

LPS-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB-responsive	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
not	NN	O	O
inhibited	NN	O	O
by	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	O
antagonist	NN	O	O
.	NN	O	O

TLR2	NN	O	B-protein
and	NN	O	O
TLR4	NN	O	B-protein
were	NN	O	O
expressed	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
of	NN	O	O
HDMEC	NN	O	B-cell_type
and	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
signal	NN	O	B-protein
transduction	NN	O	I-protein
molecule	NN	O	I-protein
in	NN	O	O
the	NN	O	O
LPS	NN	O	O
receptor	NN	O	O
complex	NN	O	O
may	NN	O	O
belong	NN	O	O
to	NN	O	O
the	NN	O	O
IL-1	NN	O	B-protein
receptor/toll-like	NN	O	O
receptor	NN	O	O
(	NN	O	O
TLR	NN	O	O
)	NN	O	O
super	NN	O	O
family	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
LPS	NN	O	O
signaling	NN	O	O
cascade	NN	O	O
uses	NN	O	O
an	NN	O	O
analogous	NN	O	O
molecular	NN	O	O
framework	NN	O	O
for	NN	O	O
signaling	NN	O	O
as	NN	O	O
IL-1	NN	O	B-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
and	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
interferon-gamma	NN	O	B-protein
receptor	NN	O	I-protein
on	NN	O	O
human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
We	NN	O	O
reported	NN	O	O
on	NN	O	O
the	NN	O	O
constitutive	NN	O	O
interferon-gamma	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
IFN-gammaR	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
on	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

But	NN	O	O
signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
IFN-gammaR	NN	O	B-protein
on	NN	O	O
eosinophils	NN	O	B-cell_type
remains	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
Jak/Stat	NN	O	B-protein
pathway	NN	O	O
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
of	NN	O	O
eosinophils	NN	O	B-cell_type
after	NN	O	O
IFN-gammaR	NN	O	B-protein
conjugation	NN	O	O
by	NN	O	O
the	NN	O	O
ligand	NN	O	O
binding	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Purified	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
eosinophils	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
at	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
for	NN	O	O
1-60	NN	O	O
min	NN	O	O
.	NN	O	O

Tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
IFN-gammaR	NN	O	B-protein
,	NN	O	O
Jak1	NN	O	B-protein
,	NN	O	O
Jak2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Stat1alpha	NN	O	B-protein
was	NN	O	O
examined	NN	O	O
by	NN	O	O
immunoblotting	NN	O	O
.	NN	O	O

Gel-shift	NN	O	O
assay	NN	O	O
was	NN	O	O
also	NN	O	O
examined	NN	O	O
to	NN	O	O
show	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
Stat1alpha-DNA	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
We	NN	O	O
show	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
to	NN	O	O
human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
initiated	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
events	NN	O	O
that	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
rapid	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
not	NN	O	O
only	NN	O	O
the	NN	O	O
IFN-gammaRalpha	NN	O	B-protein
chain	NN	O	I-protein
but	NN	O	O
also	NN	O	O
Jak1	NN	O	B-protein
,	NN	O	O
Jak2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Stat1alpha	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
IFN-gamma	NN	O	B-protein
enhanced	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
Stat1alpha	NN	O	B-protein
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
IFN-gamma	NN	O	B-protein
affects	NN	O	O
eosinophils	NN	O	B-cell_type
through	NN	O	O
its	NN	O	O
specific	NN	O	O
receptor	NN	O	O
and	NN	O	O
utilizes	NN	O	O
the	NN	O	O
Jak/Stat	NN	O	B-protein
pathway	NN	O	O
as	NN	O	O
its	NN	O	O
mode	NN	O	O
of	NN	O	O
signaling	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
E2A-HLF	NN	O	B-protein
chimeric	NN	O	I-protein
protein	NN	O	I-protein
induced	NN	O	O
T-cell	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
B-cell	NN	O	O
maturation	NN	O	O
arrest	NN	O	O
,	NN	O	O
and	NN	O	O
development	NN	O	O
of	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

The	NN	O	O
E2A-HLF	NN	O	B-DNA
fusion	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
generated	NN	O	O
by	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
17	NN	O	I-DNA
;	NN	O	I-DNA
19	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q22	NN	O	I-DNA
;	NN	O	I-DNA
p13	NN	O	I-DNA
)	NN	O	I-DNA
in	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ALL	NN	O	O
)	NN	O	O
,	NN	O	O
encodes	NN	O	O
a	NN	O	O
chimeric	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
which	NN	O	O
the	NN	O	O
trans-activating	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
E2A	NN	O	B-protein
are	NN	O	O
fused	NN	O	O
to	NN	O	O
the	NN	O	O
DNA-	NN	O	B-protein
binding	NN	O	I-protein
and	NN	O	I-protein
dimerization	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
hepatic	NN	O	B-protein
leukemic	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
HLF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
its	NN	O	O
biological	NN	O	O
role	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
expressing	NN	O	O
E2A-HLF	NN	O	B-protein
using	NN	O	O
Ig	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
direct	NN	O	O
transgene	NN	O	O
expression	NN	O	O
in	NN	O	O
cells	NN	O	O
committed	NN	O	O
to	NN	O	O
the	NN	O	O
lymphoid	NN	O	O
lineage	NN	O	O
.	NN	O	O

The	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
exhibited	NN	O	O
abnormal	NN	O	O
development	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
and	NN	O	O
spleen	NN	O	O
and	NN	O	O
were	NN	O	O
susceptible	NN	O	O
to	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
thymus	NN	O	O
contained	NN	O	O
small	NN	O	O
numbers	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
TUNEL	NN	O	O
staining	NN	O	O
showed	NN	O	O
that	NN	O	O
higher	NN	O	O
population	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
were	NN	O	O
undergoing	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
spleen	NN	O	O
exhibited	NN	O	O
a	NN	O	O
marked	NN	O	O
reduction	NN	O	O
in	NN	O	O
splenic	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
flow	NN	O	O
cytometric	NN	O	O
analyses	NN	O	O
and	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
colony	NN	O	O
formation	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
B-cell	NN	O	O
maturation	NN	O	O
was	NN	O	O
blocked	NN	O	O
at	NN	O	O
a	NN	O	O
very	NN	O	O
early	NN	O	O
developmental	NN	O	O
stage	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
E2A-HLF	NN	O	B-protein
induced	NN	O	O
T-cell	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
B-cell	NN	O	O
maturation	NN	O	O
arrest	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
susceptibility	NN	O	O
of	NN	O	O
the	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
to	NN	O	O
infection	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
immunodeficiency	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
several	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
developed	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
,	NN	O	O
classified	NN	O	O
as	NN	O	O
T-ALL	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
marker	NN	O	O
analysis	NN	O	O
and	NN	O	O
DNA	NN	O	O
rearrangements	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
an	NN	O	O
additional	NN	O	O
event	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
malignant	NN	O	O
transformation	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
expressing	NN	O	O
E2A-HLF	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
biological	NN	O	O
function	NN	O	O
of	NN	O	O
E2A-HLF	NN	O	B-protein
in	NN	O	O
lymphoid	NN	O	O
development	NN	O	O
and	NN	O	O
also	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
leukemogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RXR	NN	O	B-protein
)	NN	O	O
agonist-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
dominant-negative	NN	O	O
RXR	NN	O	B-protein
-retinoic	NN	O	O
acid	NN	O	O
receptor	NN	O	O
alpha403	NN	O	O
heterodimers	NN	O	O
is	NN	O	O
developmentally	NN	O	O
regulated	NN	O	O
during	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
multiple	NN	O	O
biologic	NN	O	O
activities	NN	O	O
of	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
are	NN	O	O
mediated	NN	O	O
through	NN	O	O
RAR	NN	O	B-protein
and	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
RXR	NN	O	I-protein
)	NN	O	I-protein
nuclear	NN	O	I-protein
receptors	NN	O	I-protein
that	NN	O	O
interact	NN	O	O
with	NN	O	O
specific	NN	O	O
DNA	NN	O	B-DNA
target	NN	O	I-DNA
sequences	NN	O	I-DNA
as	NN	O	O
heterodimers	NN	O	B-protein
(	NN	O	O
RXR-RAR	NN	O	B-protein
)	NN	O	O
or	NN	O	O
homodimers	NN	O	B-protein
(	NN	O	O
RXR-RXR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

RA	NN	O	O
receptor	NN	O	O
activation	NN	O	O
appears	NN	O	O
critical	NN	O	O
to	NN	O	O
regulating	NN	O	O
important	NN	O	O
aspects	NN	O	O
of	NN	O	O
hematopoiesis	NN	O	O
,	NN	O	O
since	NN	O	O
transducing	NN	O	O
a	NN	O	O
COOH-terminally	NN	O	B-protein
truncated	NN	O	I-protein
RARalpha	NN	O	I-protein
exhibiting	NN	O	O
dominant-negative	NN	O	O
activity	NN	O	O
(	NN	O	O
RARalpha403	NN	O	B-protein
)	NN	O	O
into	NN	O	O
normal	NN	O	O
mouse	NN	O	O
bone	NN	O	O
marrow	NN	O	O
generates	NN	O	O
hematopoietic	NN	O	B-cell_line
growth	NN	O	I-cell_line
factor-dependent	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
frozen	NN	O	O
at	NN	O	O
the	NN	O	O
multipotent	NN	O	O
progenitor	NN	O	O
(	NN	O	O
EML	NN	O	O
)	NN	O	O
or	NN	O	O
committed	NN	O	O
promyelocyte	NN	O	O
(	NN	O	O
MPRO	NN	O	O
)	NN	O	O
stages	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
relatively	NN	O	O
high	NN	O	O
,	NN	O	O
pharmacological	NN	O	O
concentrations	NN	O	O
of	NN	O	O
RA	NN	O	O
(	NN	O	O
1	NN	O	O
to	NN	O	O
10	NN	O	O
&	NN	O	O
mgr	NN	O	O
;	NN	O	O
M	NN	O	O
)	NN	O	O
overcome	NN	O	O
these	NN	O	O
differentiation	NN	O	O
blocks	NN	O	O
and	NN	O	O
induce	NN	O	O
terminal	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
MPRO	NN	O	B-cell_type
promyelocytes	NN	O	I-cell_type
while	NN	O	O
potentiating	NN	O	O
interleukin-3	NN	O	B-protein
(	NN	O	O
IL-3	NN	O	B-protein
)	NN	O	O
-induced	NN	O	O
commitment	NN	O	O
of	NN	O	O
EML	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
the	NN	O	O
granulocyte/monocyte	NN	O	O
lineage	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
utilized	NN	O	O
RXR	NN	O	B-protein
-and	NN	O	O
RAR	NN	O	B-protein
-specific	NN	O	O
agonists	NN	O	O
and	NN	O	O
antagonists	NN	O	O
to	NN	O	O
determine	NN	O	O
how	NN	O	O
RA	NN	O	O
overcomes	NN	O	O
the	NN	O	O
dominant-negative	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
truncated	NN	O	B-protein
RARalpha	NN	O	I-protein
in	NN	O	O
these	NN	O	O
different	NN	O	O
myeloid	NN	O	O
developmental	NN	O	O
stages	NN	O	O
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
an	NN	O	O
RXR	NN	O	B-protein
-specific	NN	O	O
,	NN	O	O
rather	NN	O	O
than	NN	O	O
an	NN	O	O
RAR	NN	O	B-protein
-specific	NN	O	O
,	NN	O	O
agonist	NN	O	O
induces	NN	O	O
terminal	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
MPRO	NN	O	B-cell_type
promyelocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
this	NN	O	O
differentiation	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
DNA	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
corresponding	NN	O	O
to	NN	O	O
RAR-RXR	NN	O	B-protein
heterodimers	NN	O	I-protein
rather	NN	O	O
than	NN	O	O
RXR-RXR	NN	O	B-protein
homodimers	NN	O	I-protein
.	NN	O	O

This	NN	O	O
RXR	NN	O	B-protein
agonist	NN	O	O
activity	NN	O	O
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
RAR	NN	O	B-protein
-specific	NN	O	O
antagonists	NN	O	O
,	NN	O	O
suggesting	NN	O	O
extensive	NN	O	O
cross-talk	NN	O	O
between	NN	O	O
the	NN	O	O
partners	NN	O	O
of	NN	O	O
the	NN	O	O
RXR	NN	O	B-protein
-RARalpha403	NN	O	I-protein
heterodimer	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
more	NN	O	O
immature	NN	O	O
,	NN	O	O
multipotent	NN	O	O
EML	NN	O	B-cell_type
cells	NN	O	I-cell_type
we	NN	O	O
observed	NN	O	O
that	NN	O	O
this	NN	O	O
RXR	NN	O	B-protein
-specific	NN	O	O
agonist	NN	O	O
is	NN	O	O
inactive	NN	O	O
either	NN	O	O
in	NN	O	O
potentiating	NN	O	O
IL-3	NN	O	B-protein
-mediated	NN	O	O
commitment	NN	O	O
of	NN	O	O
EML	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
the	NN	O	O
granulocyte	NN	O	B-cell_type
lineage	NN	O	I-cell_type
or	NN	O	O
in	NN	O	O
transactivating	NN	O	O
RAR-RXR	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

RA	NN	O	O
-triggered	NN	O	O
GALdbd-	NN	O	O
RARalpha	NN	O	B-protein
hybrid	NN	O	O
activity	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
multipotent	NN	O	O
EML	NN	O	B-cell_type
cells	NN	O	I-cell_type
harbor	NN	O	O
substantial	NN	O	O
nuclear	NN	O	O
hormone	NN	O	O
receptor	NN	O	O
coactivator	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
histone	NN	O	B-protein
deacetylase	NN	O	I-protein
(	NN	O	I-protein
HDAC	NN	O	I-protein
)	NN	O	I-protein
inhibitor	NN	O	I-protein
trichostatin	NN	O	O
A	NN	O	O
readily	NN	O	O
activates	NN	O	O
an	NN	O	O
RXR-RAR	NN	O	B-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
multipotent	NN	O	O
EML	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
committed	NN	O	O
MPRO	NN	O	B-cell_type
promyelocytes	NN	O	I-cell_type
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
differences	NN	O	O
in	NN	O	O
HDAC-containing	NN	O	B-protein
repressor	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
these	NN	O	O
two	NN	O	O
closely	NN	O	O
related	NN	O	O
but	NN	O	O
distinct	NN	O	O
hematopoietic	NN	O	B-cell_type
lineages	NN	O	I-cell_type
might	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
differential	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
RXR-RARalpha403	NN	O	B-protein
heterodimers	NN	O	I-protein
that	NN	O	O
we	NN	O	O
observed	NN	O	O
at	NN	O	O
these	NN	O	O
different	NN	O	O
stages	NN	O	O
of	NN	O	O
myeloid	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
functional	NN	O	O
synergy	NN	O	O
between	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
involves	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
augmentation	NN	O	O
of	NN	O	O
STAT	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

IL-12	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
can	NN	O	O
stimulate	NN	O	O
mitogen-	NN	O	B-cell_type
or	NN	O	I-cell_type
CD3-activated	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
proliferate	NN	O	O
,	NN	O	O
produce	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
kill	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
magnitude	NN	O	O
of	NN	O	O
these	NN	O	O
functional	NN	O	O
responses	NN	O	O
is	NN	O	O
greatly	NN	O	O
augmented	NN	O	O
when	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
largely	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
without	NN	O	O
prior	NN	O	O
activation	NN	O	O
,	NN	O	O
a	NN	O	O
small	NN	O	O
subset	NN	O	O
of	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
CD8+CD18bright	NN	O	B-cell_type
)	NN	O	O
is	NN	O	O
strongly	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
functional	NN	O	O
synergy	NN	O	O
between	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
CD8+CD18bright	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
stress	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
p38	NN	O	B-protein
mitogen-activated	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MAP	NN	O	I-protein
)	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
stress-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
SAPK	NN	O	I-protein
)	NN	O	I-protein
/Jun	NN	O	I-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

The	NN	O	O
functional	NN	O	O
synergy	NN	O	O
between	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
is	NN	O	O
also	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
prominent	NN	O	O
increase	NN	O	O
in	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
serine	NN	O	O
phosphorylation	NN	O	O
over	NN	O	O
that	NN	O	O
observed	NN	O	O
with	NN	O	O
IL-12	NN	O	B-protein
or	NN	O	O
IL-2	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
STAT	NN	O	B-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
is	NN	O	O
not	NN	O	O
augmented	NN	O	O
over	NN	O	O
that	NN	O	O
seen	NN	O	O
with	NN	O	O
either	NN	O	O
cytokine	NN	O	O
alone	NN	O	O
.	NN	O	O

A	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
p38	NN	O	O
MAP	NN	O	O
kinase	NN	O	O
completely	NN	O	O
inhibits	NN	O	O
the	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
induced	NN	O	O
by	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
abrogates	NN	O	O
the	NN	O	O
functional	NN	O	O
synergy	NN	O	O
between	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
without	NN	O	O
affecting	NN	O	O
STAT	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
p38	NN	O	O
MAP	NN	O	O
kinase	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
STAT	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
optimal	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
IL-12	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
may	NN	O	O
depend	NN	O	O
on	NN	O	O
an	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
p38	NN	O	B-protein
MAP	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
Janus	NN	O	B-protein
kinase/STAT	NN	O	I-protein
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
Mycobacterium	NN	O	O
tuberculosis	NN	O	O
-induced	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
.	NN	O	O

Soluble	NN	O	O
interleukin-2	NN	O	B-protein
receptor-alpha	NN	O	I-protein
(	NN	O	O
IL-2Ralpha	NN	O	B-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
increased	NN	O	O
in	NN	O	O
the	NN	O	O
sera	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
advanced	NN	O	O
tuberculosis	NN	O	O
,	NN	O	O
and	NN	O	O
levels	NN	O	O
decline	NN	O	O
after	NN	O	O
therapy	NN	O	O
in	NN	O	O
accordance	NN	O	O
with	NN	O	O
improvement	NN	O	O
of	NN	O	O
radiologic	NN	O	O
findings	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2Ralpha	NN	O	B-protein
in	NN	O	O
bronchoalveolar	NN	O	B-cell_type
lavage	NN	O	I-cell_type
(	NN	O	I-cell_type
BAL	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
active	NN	O	O
pulmonary	NN	O	O
tuberculosis	NN	O	O
,	NN	O	O
and	NN	O	O
evaluated	NN	O	O
the	NN	O	O
mechanism	NN	O	O
Mycobacterium	NN	O	O
tuberculosis	NN	O	O
induces	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2Ralpha	NN	O	B-protein
using	NN	O	O
the	NN	O	O
THP-1	NN	O	B-cell_line
mononuclear	NN	O	I-cell_line
phagocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
found	NN	O	O
IL-2Ralpha	NN	O	B-protein
expression	NN	O	O
to	NN	O	O
be	NN	O	O
increased	NN	O	O
in	NN	O	O
BAL	NN	O	O
cells	NN	O	O
from	NN	O	O
involved	NN	O	O
sites	NN	O	O
of	NN	O	O
active	NN	O	O
pulmonary	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
alpha-chain	NN	O	B-protein
of	NN	O	O
IL-2Ralpha	NN	O	B-protein
on	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
(	NN	O	O
PBM	NN	O	B-cell_type
)	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
evaluation	NN	O	O
.	NN	O	O

Northern	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
increased	NN	O	O
IL-2Ralpha	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
after	NN	O	O
stimulation	NN	O	O
with	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
which	NN	O	O
was	NN	O	O
further	NN	O	O
induced	NN	O	O
by	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
IL-2Ralpha	NN	O	B-DNA
promoter	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
(	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
transcriptionally	NN	O	O
induced	NN	O	O
by	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
and	NN	O	O
this	NN	O	O
NF-kappaB	NN	O	B-protein
site	NN	O	O
could	NN	O	O
confer	NN	O	O
inducibility	NN	O	O
to	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
herpes	NN	O	I-DNA
thymidine	NN	O	I-DNA
kinase	NN	O	I-DNA
(	NN	O	I-DNA
TK	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSAs	NN	O	O
)	NN	O	O
revealed	NN	O	O
specific	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	O
protein	NN	O	O
to	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-DNA
site	NN	O	I-DNA
upon	NN	O	O
induction	NN	O	O
with	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

Using	NN	O	O
antibodies	NN	O	O
against	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
subunits	NN	O	I-protein
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
EMSAs	NN	O	O
,	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
both	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
proteins	NN	O	I-protein
was	NN	O	O
further	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

Functional	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2Ralpha	NN	O	B-protein
on	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
in	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
infection	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
immunomodulatory	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
host	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
3'-untranslated	NN	O	B-DNA
region	NN	O	I-DNA
polymorphisms	NN	O	O
:	NN	O	O
lack	NN	O	O
of	NN	O	O
effect	NN	O	O
on	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
.	NN	O	O

Allelic	NN	O	O
variation	NN	O	O
at	NN	O	O
the	NN	O	O
3'-end	NN	O	O
of	NN	O	O
the	NN	O	O
vitamin	NN	O	B-DNA
D	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
VDR	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
3-5-fold	NN	O	O
increased	NN	O	O
risk	NN	O	O
of	NN	O	O
developing	NN	O	O
prostate	NN	O	O
cancer	NN	O	O
and	NN	O	O
with	NN	O	O
differences	NN	O	O
in	NN	O	O
bone	NN	O	O
mineralization	NN	O	O
.	NN	O	O

This	NN	O	O
genetic	NN	O	O
diversity	NN	O	O
does	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
VDR	NN	O	B-protein
protein	NN	O	I-protein
structurally	NN	O	O
,	NN	O	O
but	NN	O	O
instead	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
marker	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
other	NN	O	O
,	NN	O	O
nearby	NN	O	O
polymorphisms	NN	O	O
that	NN	O	O
influence	NN	O	O
message	NN	O	O
stability	NN	O	O
or	NN	O	O
translation	NN	O	O
.	NN	O	O

The	NN	O	O
work	NN	O	O
reported	NN	O	O
here	NN	O	O
was	NN	O	O
instigated	NN	O	O
to	NN	O	O
identify	NN	O	O
additional	NN	O	O
VDR	NN	O	B-protein
3'-UTR	NN	O	B-DNA
polymorphisms	NN	O	O
that	NN	O	O
may	NN	O	O
have	NN	O	O
functional	NN	O	O
significance	NN	O	O
and	NN	O	O
to	NN	O	O
then	NN	O	O
test	NN	O	O
whether	NN	O	O
these	NN	O	O
genetic	NN	O	O
variants	NN	O	O
alter	NN	O	O
message	NN	O	O
stability	NN	O	O
.	NN	O	O

Initially	NN	O	O
,	NN	O	O
four	NN	O	O
novel	NN	O	O
,	NN	O	O
frequently	NN	O	O
occurring	NN	O	O
sequence	NN	O	B-DNA
variants	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
that	NN	O	O
associated	NN	O	O
with	NN	O	O
two	NN	O	O
common	NN	O	O
haplotypes	NN	O	O
that	NN	O	O
were	NN	O	O
described	NN	O	O
previously	NN	O	O
.	NN	O	O

These	NN	O	O
common	NN	O	O
sequence	NN	O	O
variants	NN	O	O
were	NN	O	O
not	NN	O	O
found	NN	O	O
within	NN	O	O
three	NN	O	O
message-destabilizing	NN	O	B-DNA
elements	NN	O	I-DNA
that	NN	O	O
we	NN	O	O
mapped	NN	O	O
within	NN	O	O
the	NN	O	O
3'-UTR	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
vitamin	NN	O	B-RNA
D	NN	O	I-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
two	NN	O	O
VDR	NN	O	B-DNA
3'-UTR	NN	O	I-DNA
haplotypes	NN	O	O
conferred	NN	O	O
an	NN	O	O
identical	NN	O	O
half-life	NN	O	O
on	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
beta-globin	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
assay	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
conclude	NN	O	O
that	NN	O	O
common	NN	O	O
polymorphisms	NN	O	O
within	NN	O	O
the	NN	O	O
VDR	NN	O	B-DNA
3'-UTR	NN	O	I-DNA
do	NN	O	O
not	NN	O	O
influence	NN	O	O
message	NN	O	O
stability	NN	O	O
.	NN	O	O

-DOCSTART-	O

IL-2	NN	O	B-protein
-mediated	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
does	NN	O	O
not	NN	O	O
require	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
or	NN	O	O
activating	NN	O	O
protein-1	NN	O	O
activation	NN	O	O
in	NN	O	O
primary	NN	O	O
human	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
IL-2	NN	O	B-protein
growth	NN	O	O
hormone	NN	O	O
is	NN	O	O
the	NN	O	O
major	NN	O	O
growth	NN	O	O
factor	NN	O	O
of	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
during	NN	O	O
a	NN	O	O
developing	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

IL-2	NN	O	B-protein
is	NN	O	O
required	NN	O	O
not	NN	O	O
only	NN	O	O
for	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
but	NN	O	O
also	NN	O	O
to	NN	O	O
protect	NN	O	O
Ag-activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

In	NN	O	O
several	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and/or	NN	O	O
activating	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
be	NN	O	O
extremely	NN	O	O
important	NN	O	O
in	NN	O	O
blocking	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
either	NN	O	O
or	NN	O	O
both	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
cell	NN	O	O
survival	NN	O	O
or	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
responsive	NN	O	O
to	NN	O	O
the	NN	O	O
growth	NN	O	O
factor	NN	O	O
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
and	NN	O	O
AP-1	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
current	NN	O	O
study	NN	O	O
clearly	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
does	NN	O	O
not	NN	O	O
induce	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	O
degradation	NN	O	O
or	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
respond	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
by	NN	O	O
entering	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
and	NN	O	O
avoiding	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
neither	NN	O	O
activates	NN	O	O
JNK	NN	O	B-protein
nor	NN	O	O
increases	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
to	NN	O	O
a	NN	O	O
consensus	NN	O	O
o-tetradecanoylphorbol	NN	O	B-DNA
13-acetate	NN	O	I-DNA
(	NN	O	I-DNA
TPA	NN	O	I-DNA
)	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
the	NN	O	O
growth	NN	O	B-protein
factor	NN	O	I-protein
does	NN	O	O
induce	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
as	NN	O	O
has	NN	O	O
been	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
neither	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
nor	NN	O	O
AP-1	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
-mediated	NN	O	O
survival	NN	O	O
or	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
in	NN	O	O
activated	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Estrone	NN	O	O
potentiates	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
:	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
17	NN	O	B-protein
beta-hydroxysteroid	NN	O	I-protein
dehydrogenase	NN	O	I-protein
in	NN	O	O
modulating	NN	O	O
hemopoiesis	NN	O	O
.	NN	O	O

Hormones	NN	O	O
such	NN	O	O
as	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxy	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
D3	NN	O	O
)	NN	O	O
,	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
,	NN	O	O
and	NN	O	O
9-cis	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
stimulate	NN	O	O
differentiation	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
their	NN	O	O
interaction	NN	O	O
with	NN	O	O
specific	NN	O	O
hormone	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
cells	NN	O	O
to	NN	O	O
these	NN	O	O
agents	NN	O	O
is	NN	O	O
not	NN	O	O
merely	NN	O	O
governed	NN	O	O
by	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
their	NN	O	O
receptors	NN	O	O
and	NN	O	O
the	NN	O	O
availability	NN	O	O
of	NN	O	O
ligand	NN	O	O
to	NN	O	O
bind	NN	O	O
them	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
from	NN	O	O
our	NN	O	O
group	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
D3	NN	O	O
and	NN	O	O
retinoids	NN	O	O
on	NN	O	O
myelopoiesis	NN	O	O
also	NN	O	O
are	NN	O	O
influenced	NN	O	O
by	NN	O	O
endogenous	NN	O	O
mechanisms	NN	O	O
involving	NN	O	O
other	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
local	NN	O	O
estrogen	NN	O	O
metabolism	NN	O	O
on	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
normal	NN	O	O
primitive	NN	O	O
myeloid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Quantitative	NN	O	O
thin-layer	NN	O	O
chromatography	NN	O	O
(	NN	O	O
TLC	NN	O	O
)	NN	O	O
analyses	NN	O	O
showed	NN	O	O
that	NN	O	O
HL60	NN	O	B-cell_line
and	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
able	NN	O	O
to	NN	O	O
generate	NN	O	O
estrone	NN	O	O
(	NN	O	O
E1	NN	O	O
)	NN	O	O
from	NN	O	O
estradiol	NN	O	O
(	NN	O	O
E2	NN	O	O
)	NN	O	O
.	NN	O	O

Neither	NN	O	O
cell	NN	O	O
population	NN	O	O
generated	NN	O	O
significant	NN	O	O
amounts	NN	O	O
of	NN	O	O
E2	NN	O	O
from	NN	O	O
E1	NN	O	O
.	NN	O	O

Reverse	NN	O	O
transcriptase	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
and	NN	O	O
Northern	NN	O	O
analyses	NN	O	O
confirmed	NN	O	O
that	NN	O	O
normal	NN	O	O
and	NN	O	O
leukemic	NN	O	O
myeloid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
expressed	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
the	NN	O	O
type	NN	O	O
I	NN	O	O
and	NN	O	O
IV	NN	O	O
isoforms	NN	O	O
of	NN	O	O
17	NN	O	B-protein
beta-hydroxysteroid	NN	O	I-protein
dehydrogenase	NN	O	I-protein
.	NN	O	O

Conversion	NN	O	O
of	NN	O	O
E2	NN	O	O
to	NN	O	O
E1	NN	O	O
was	NN	O	O
upregulated	NN	O	O
within	NN	O	O
24	NN	O	O
hours	NN	O	O
when	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
treated	NN	O	O
with	NN	O	O
either	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
or	NN	O	O
D3	NN	O	O
at	NN	O	O
doses	NN	O	O
that	NN	O	O
induce	NN	O	O
their	NN	O	O
differentiation	NN	O	O
toward	NN	O	O
neutrophils	NN	O	B-cell_type
or	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
D3-induced	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
of	NN	O	O
normal	NN	O	O
myeloid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
associated	NN	O	O
with	NN	O	O
increased	NN	O	O
capacity	NN	O	O
to	NN	O	O
generate	NN	O	O
E1	NN	O	O
from	NN	O	O
E2	NN	O	O
.	NN	O	O

When	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
normal	NN	O	O
myeloid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
exogenous	NN	O	O
E1	NN	O	O
they	NN	O	O
became	NN	O	O
more	NN	O	O
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
differentiation-inducing	NN	O	O
effects	NN	O	O
of	NN	O	O
D3	NN	O	O
.	NN	O	O

Data	NN	O	O
presented	NN	O	O
provide	NN	O	O
further	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
local	NN	O	O
modulation	NN	O	O
of	NN	O	O
myelopoiesis	NN	O	O
by	NN	O	O
intracrine	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
local	NN	O	O
metabolism	NN	O	O
of	NN	O	O
steroids	NN	O	O
by	NN	O	O
normal	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
leukemic	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
influences	NN	O	O
their	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
D3	NN	O	O
and	NN	O	O
retinoids	NN	O	O
.	NN	O	O

-DOCSTART-	O

Phosphorylation	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
inhibits	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

TRAF2	NN	O	B-protein
is	NN	O	O
a	NN	O	O
signal	NN	O	B-protein
transducing	NN	O	I-protein
adaptor	NN	O	I-protein
molecule	NN	O	I-protein
which	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
phosphorylated	NN	O	O
,	NN	O	O
predominantly	NN	O	O
on	NN	O	O
serine	NN	O	O
residues	NN	O	O
,	NN	O	O
when	NN	O	O
transiently	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
293	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
phosphorylation	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
signaling	NN	O	O
events	NN	O	O
that	NN	O	O
are	NN	O	O
activated	NN	O	O
by	NN	O	O
TRAF2	NN	O	B-protein
under	NN	O	O
these	NN	O	O
circumstances	NN	O	O
,	NN	O	O
since	NN	O	O
two	NN	O	O
nonfunctional	NN	O	O
mutants	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
phosphorylated	NN	O	O
significantly	NN	O	O
less	NN	O	O
than	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
status	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
had	NN	O	O
significant	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
CD40	NN	O	B-protein
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
our	NN	O	O
observations	NN	O	O
that	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
interacted	NN	O	O
preferentially	NN	O	O
with	NN	O	O
underphosphorylated	NN	O	O
TRAF2	NN	O	B-protein
and	NN	O	O
that	NN	O	O
phosphatase	NN	O	B-protein
treatment	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
to	NN	O	O
CD40	NN	O	B-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
from	NN	O	O
these	NN	O	O
studies	NN	O	O
that	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
is	NN	O	O
likely	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
signaling	NN	O	O
by	NN	O	O
virtue	NN	O	O
of	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
influence	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
-TRAF2	NN	O	B-protein
interaction	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
direct	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
adaptor	NN	O	B-protein
protein	NN	O	I-protein
Cbl-b	NN	O	B-protein
and	NN	O	O
the	NN	O	O
kinase	NN	O	B-protein
zap-70	NN	O	B-protein
induces	NN	O	O
a	NN	O	O
positive	NN	O	O
signal	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Engagement	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
TCR	NN	O	I-protein
)	NN	O	I-protein
-CD3	NN	O	I-protein
complex	NN	O	I-protein
induces	NN	O	O
a	NN	O	O
rapid	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
Src-family	NN	O	B-protein
and	NN	O	O
Syk/Zap-70-family	NN	O	B-protein
kinases	NN	O	I-protein
[	NN	O	O
1	NN	O	O
]	NN	O	O
[	NN	O	O
2	NN	O	O
]	NN	O	O
.	NN	O	O

These	NN	O	O
activated	NN	O	B-protein
kinases	NN	O	I-protein
then	NN	O	O
induce	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
multiple	NN	O	B-protein
intracellular	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
eventually	NN	O	O
leading	NN	O	O
to	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
prominent	NN	O	O
substrates	NN	O	O
for	NN	O	O
these	NN	O	O
kinases	NN	O	B-protein
is	NN	O	O
the	NN	O	O
adaptor	NN	O	B-protein
protein	NN	O	I-protein
Cbl	NN	O	O
[	NN	O	O
3	NN	O	O
]	NN	O	O
and	NN	O	O
recent	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
Cbl	NN	O	B-protein
negatively	NN	O	O
regulates	NN	O	O
upstream	NN	O	B-protein
kinases	NN	O	I-protein
such	NN	O	O
as	NN	O	O
Syk	NN	O	B-protein
and	NN	O	O
Zap-70	NN	O	B-protein
[	NN	O	O
4	NN	O	O
]	NN	O	O
[	NN	O	O
5	NN	O	O
]	NN	O	O
.	NN	O	O

Cbl-b	NN	O	B-protein
,	NN	O	O
a	NN	O	O
homologue	NN	O	O
of	NN	O	O
Cbl	NN	O	B-protein
,	NN	O	O
is	NN	O	O
widely	NN	O	O
expressed	NN	O	O
in	NN	O	O
many	NN	O	O
tissues	NN	O	O
and	NN	O	O
cells	NN	O	O
including	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
[	NN	O	O
6	NN	O	O
]	NN	O	O
[	NN	O	O
7	NN	O	O
]	NN	O	O
.	NN	O	O

Cbl-b	NN	O	B-protein
undergoes	NN	O	O
rapid	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
cytokine	NN	O	O
receptors	NN	O	O
[	NN	O	O
8	NN	O	O
]	NN	O	O
[	NN	O	O
9	NN	O	O
]	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
Cbl-b	NN	O	B-protein
is	NN	O	O
unclear	NN	O	O
,	NN	O	O
however	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Cbl-b	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
induced	NN	O	O
the	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
.	NN	O	O

A	NN	O	O
loss-of-function	NN	O	O
mutation	NN	O	O
in	NN	O	O
Cbl-b	NN	O	B-protein
disrupted	NN	O	O
the	NN	O	O
interaction	NN	O	O
between	NN	O	O
Cbl-b	NN	O	B-protein
and	NN	O	O
Zap-70	NN	O	B-protein
and	NN	O	O
nearly	NN	O	O
completely	NN	O	O
abrogated	NN	O	O
the	NN	O	O
Cbl-b	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

Unlike	NN	O	O
the	NN	O	O
proposed	NN	O	O
role	NN	O	O
of	NN	O	O
Cbl	NN	O	B-protein
as	NN	O	O
a	NN	O	O
negative	NN	O	O
regulator	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
Cbl	NN	O	B-protein
homologue	NN	O	O
Cbl-b	NN	O	B-protein
has	NN	O	O
a	NN	O	O
positive	NN	O	O
role	NN	O	O
in	NN	O	O
T-cell	NN	O	O
signaling	NN	O	O
,	NN	O	O
most	NN	O	O
likely	NN	O	O
via	NN	O	O
a	NN	O	O
direct	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
upstream	NN	O	B-protein
kinase	NN	O	I-protein
Zap-70	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Grf40	NN	O	B-protein
,	NN	O	O
A	NN	O	O
novel	NN	O	O
Grb2	NN	O	B-protein
family	NN	O	I-protein
member	NN	O	I-protein
,	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
signaling	NN	O	O
through	NN	O	O
interaction	NN	O	O
with	NN	O	O
SLP-76	NN	O	B-protein
and	NN	O	O
LAT	NN	O	B-protein
.	NN	O	O

We	NN	O	O
molecularly	NN	O	O
cloned	NN	O	O
a	NN	O	O
new	NN	O	O
Grb2	NN	O	B-protein
family	NN	O	I-protein
member	NN	O	I-protein
,	NN	O	O
named	NN	O	O
Grf40	NN	O	B-protein
,	NN	O	O
containing	NN	O	O
the	NN	O	O
common	NN	O	O
SH3-SH2-SH3	NN	O	B-protein
motif	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
Grf40	NN	O	B-protein
is	NN	O	O
predominant	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
particularly	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Grf40	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
domain-containing	NN	O	I-protein
leukocyte	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
76	NN	O	O
kD	NN	O	O
(	NN	O	O
SLP-76	NN	O	B-protein
)	NN	O	O
via	NN	O	O
its	NN	O	O
SH3	NN	O	O
domain	NN	O	O
more	NN	O	O
tightly	NN	O	O
than	NN	O	O
Grb2	NN	O	B-protein
.	NN	O	O

Incidentally	NN	O	O
,	NN	O	O
Grf40	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
linker	NN	O	B-protein
for	NN	O	I-protein
activation	NN	O	I-protein
of	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
LAT	NN	O	B-protein
)	NN	O	O
possibly	NN	O	O
via	NN	O	O
its	NN	O	O
SH2	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
wild-type	NN	O	B-protein
Grf40	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
induced	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
of	NN	O	O
SLP-76-dependent	NN	O	B-DNA
interleukin	NN	O	I-DNA
(	NN	O	I-DNA
IL	NN	O	I-DNA
)	NN	O	I-DNA
-2	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
upon	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
stimulation	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
COOH-terminal	NN	O	B-protein
SH3-deleted	NN	O	I-protein
Grf40	NN	O	I-protein
mutant	NN	O	I-protein
lacked	NN	O	O
any	NN	O	O
recognizable	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
SH2-deleted	NN	O	B-protein
Grf40	NN	O	I-protein
mutant	NN	O	I-protein
led	NN	O	O
to	NN	O	O
a	NN	O	O
marked	NN	O	O
inhibition	NN	O	O
of	NN	O	O
these	NN	O	O
regulatory	NN	O	O
activities	NN	O	O
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
apparently	NN	O	O
stronger	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
SH2-deleted	NN	O	B-protein
Grb2	NN	O	I-protein
mutant	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
Grf40	NN	O	B-protein
is	NN	O	O
an	NN	O	O
adaptor	NN	O	B-protein
molecule	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
through	NN	O	O
a	NN	O	O
more	NN	O	O
efficient	NN	O	O
interaction	NN	O	O
than	NN	O	O
Grb2	NN	O	B-protein
with	NN	O	O
SLP-76	NN	O	B-protein
and	NN	O	O
LAT	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Modulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
and	NN	O	O
tumor	NN	O	O
growth	NN	O	O
by	NN	O	O
activated	NN	O	O
Ras	NN	O	B-protein
.	NN	O	O

As	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
its	NN	O	O
transforming	NN	O	O
abilities	NN	O	O
,	NN	O	O
activated	NN	O	B-protein
Ras	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
great	NN	O	O
number	NN	O	O
of	NN	O	O
cancers	NN	O	O
.	NN	O	O

The	NN	O	O
ras	NN	O	O
mutation	NN	O	O
frequency	NN	O	O
varies	NN	O	O
between	NN	O	O
95	NN	O	O
%	NN	O	O
in	NN	O	O
pancreatic	NN	O	O
cancer	NN	O	O
and	NN	O	O
5	NN	O	O
%	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

In	NN	O	O
leukemia	NN	O	O
,	NN	O	O
the	NN	O	O
highest	NN	O	O
frequency	NN	O	O
(	NN	O	O
30	NN	O	O
%	NN	O	O
)	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
ras	NN	O	O
mutations	NN	O	O
has	NN	O	O
been	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
poor	NN	O	O
prognosis	NN	O	O
and	NN	O	O
negative	NN	O	O
clinical	NN	O	O
outcome	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
mutated	NN	O	B-protein
Ras	NN	O	I-protein
activates	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
which	NN	O	O
favor	NN	O	O
tumor	NN	O	O
growth	NN	O	O
,	NN	O	O
enhance	NN	O	O
the	NN	O	O
metastatic	NN	O	O
capacity	NN	O	O
of	NN	O	O
tumors	NN	O	O
or	NN	O	O
modulate	NN	O	O
tumor-specific	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

Several	NN	O	O
new	NN	O	O
functions	NN	O	O
of	NN	O	O
Ras	NN	O	B-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
downregulation	NN	O	O
of	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
upregulation	NN	O	O
of	NN	O	O
certain	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
degradative	NN	O	B-protein
enzymes	NN	O	I-protein
have	NN	O	O
been	NN	O	O
uncovered	NN	O	O
in	NN	O	O
the	NN	O	O
last	NN	O	O
decade	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
mutated	NN	O	B-protein
Ras	NN	O	I-protein
can	NN	O	O
also	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
primary	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
immunotherapy	NN	O	O
or	NN	O	O
drug	NN	O	O
therapy	NN	O	O
.	NN	O	O

This	NN	O	O
review	NN	O	O
will	NN	O	O
discuss	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
Ras	NN	O	B-protein
expressing	NN	O	O
tumors	NN	O	O
are	NN	O	O
able	NN	O	O
to	NN	O	O
evade	NN	O	O
destruction	NN	O	O
by	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
and	NN	O	O
enhance	NN	O	O
their	NN	O	O
growth	NN	O	O
and	NN	O	O
metastatic	NN	O	O
potential	NN	O	O
.	NN	O	O

It	NN	O	O
will	NN	O	O
further	NN	O	O
elaborate	NN	O	O
on	NN	O	O
the	NN	O	O
attempts	NN	O	O
to	NN	O	O
develop	NN	O	O
successful	NN	O	O
immunotherapy	NN	O	O
and	NN	O	O
drug	NN	O	O
therapy	NN	O	O
targeting	NN	O	O
Ras	NN	O	B-protein
expressing	NN	O	O
tumors	NN	O	O
.	NN	O	O

-DOCSTART-	O

Modulation	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
complexes	NN	O	I-protein
during	NN	O	O
G0	NN	O	O
to	NN	O	O
S	NN	O	O
phase	NN	O	O
transition	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
primary	NN	O	I-cell_type
B-lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
pocket	NN	O	B-protein
protein-E2F	NN	O	I-protein
complexes	NN	O	I-protein
are	NN	O	O
convergence	NN	O	O
points	NN	O	O
for	NN	O	O
cell	NN	O	O
cycle	NN	O	O
signaling	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
and	NN	O	O
monitored	NN	O	O
the	NN	O	O
pocket	NN	O	B-protein
protein-E2F	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
primary	NN	O	I-cell_type
B-lymphocytes	NN	O	I-cell_type
after	NN	O	O
activation	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
previous	NN	O	O
data	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
and	NN	O	I-cell_type
mouse	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
and	NN	O	O
T-lymphocytes	NN	O	B-cell_type
,	NN	O	O
E2F4	NN	O	B-protein
and	NN	O	O
DP1	NN	O	B-protein
form	NN	O	O
the	NN	O	O
predominant	NN	O	O
E2F	NN	O	B-protein
heterodimers	NN	O	I-protein
both	NN	O	O
in	NN	O	O
G0	NN	O	O
and	NN	O	O
G1	NN	O	O
phases	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
B-lymphocyte	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
whereas	NN	O	O
E2F1	NN	O	B-protein
and	NN	O	I-protein
-3	NN	O	I-protein
are	NN	O	O
first	NN	O	O
detected	NN	O	O
in	NN	O	O
late	NN	O	O
G1	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
expression	NN	O	O
levels	NN	O	O
increase	NN	O	O
towards	NN	O	O
S	NN	O	O
phase	NN	O	O
.	NN	O	O

Intriguingly	NN	O	O
,	NN	O	O
the	NN	O	O
major	NN	O	B-protein
E2F	NN	O	I-protein
complex	NN	O	I-protein
that	NN	O	O
we	NN	O	O
detected	NN	O	O
in	NN	O	O
quiescent	NN	O	B-cell_type
human	NN	O	I-cell_type
B-lymphocytes	NN	O	I-cell_type
is	NN	O	O
consisted	NN	O	O
of	NN	O	O
pRB	NN	O	B-protein
,	NN	O	O
E2F4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
DP1	NN	O	B-protein
.	NN	O	O

Though	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
DP1	NN	O	B-protein
and	NN	O	I-protein
-2	NN	O	I-protein
increase	NN	O	O
when	NN	O	O
cells	NN	O	O
progress	NN	O	O
from	NN	O	O
G0	NN	O	O
to	NN	O	O
S	NN	O	O
,	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
DP1	NN	O	B-protein
to	NN	O	O
DP2	NN	O	B-protein
remains	NN	O	O
relatively	NN	O	O
constant	NN	O	O
during	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
observed	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
of	NN	O	O
the	NN	O	O
predominant	NN	O	O
E2F	NN	O	B-protein
components	NN	O	I-protein
,	NN	O	O
DP1	NN	O	B-protein
and	NN	O	O
E2F4	NN	O	B-protein
,	NN	O	O
as	NN	O	O
B-lymphocytes	NN	O	B-cell_type
progressed	NN	O	O
from	NN	O	O
G0	NN	O	O
into	NN	O	O
early	NN	O	O
G1	NN	O	O
.	NN	O	O

This	NN	O	O
increase	NN	O	O
in	NN	O	O
mobility	NN	O	O
was	NN	O	O
attributable	NN	O	O
to	NN	O	O
dephosphorylation	NN	O	O
,	NN	O	O
as	NN	O	O
lambda	NN	O	O
phosphatase	NN	O	O
treatment	NN	O	O
could	NN	O	O
convert	NN	O	O
the	NN	O	O
slower	NN	O	B-protein
migrating	NN	O	I-protein
forms	NN	O	I-protein
into	NN	O	O
the	NN	O	O
corresponding	NN	O	B-protein
faster	NN	O	I-protein
mobility	NN	O	I-protein
forms	NN	O	I-protein
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
this	NN	O	O
change	NN	O	O
in	NN	O	O
phosphorylation	NN	O	O
status	NN	O	O
correlates	NN	O	O
with	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

This	NN	O	O
modulation	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
DP1	NN	O	B-protein
and	NN	O	O
E2F4	NN	O	B-protein
could	NN	O	O
help	NN	O	O
to	NN	O	O
explain	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
in	NN	O	O
vivo	NN	O	O
DNA	NN	O	O
footprinting	NN	O	O
in	NN	O	O
late	NN	O	O
G1	NN	O	O
and	NN	O	O
S	NN	O	O
phases	NN	O	O
of	NN	O	O
gene	NN	O	B-DNA
promoters	NN	O	I-DNA
negatively	NN	O	O
regulated	NN	O	O
through	NN	O	O
E2F	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
suggests	NN	O	O
a	NN	O	O
novel	NN	O	O
mechanism	NN	O	O
for	NN	O	O
controlling	NN	O	O
E2F	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
during	NN	O	O
the	NN	O	O
transition	NN	O	O
from	NN	O	O
quiescence	NN	O	O
to	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Engagement	NN	O	O
of	NN	O	O
natural	NN	O	O
cytotoxicity	NN	O	O
programs	NN	O	O
regulates	NN	O	O
AP-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
the	NN	O	O
NKL	NN	O	B-cell_line
human	NN	O	I-cell_line
NK	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

NK	NN	O	O
cell	NN	O	O
cytotoxicity	NN	O	O
is	NN	O	O
a	NN	O	O
fast	NN	O	O
and	NN	O	O
efficient	NN	O	O
mechanism	NN	O	O
of	NN	O	O
target	NN	O	O
cell	NN	O	O
lysis	NN	O	O
.	NN	O	O

Using	NN	O	O
transcription	NN	O	O
analysis	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
multiplex	NN	O	O
messenger	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
natural	NN	O	O
cytotoxicity	NN	O	O
exerted	NN	O	O
by	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
NKL	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
correlates	NN	O	O
with	NN	O	O
mRNA	NN	O	O
accumulation	NN	O	O
of	NN	O	O
very	NN	O	B-protein
early	NN	O	I-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
AP	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
transcription	NN	O	B-DNA
factor	NN	O	I-DNA
genes	NN	O	I-DNA
such	NN	O	O
as	NN	O	O
JunB	NN	O	B-DNA
,	NN	O	O
FosB	NN	O	B-DNA
and	NN	O	O
c-Fos	NN	O	B-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
DNA-binding	NN	O	O
activities	NN	O	O
of	NN	O	O
Jun-Fos	NN	O	B-protein
heterodimers	NN	O	I-protein
were	NN	O	O
observed	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
during	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
natural	NN	O	O
cytotoxicity	NN	O	O
.	NN	O	O

Interaction	NN	O	O
between	NN	O	O
immunoglobulin-like	NN	O	B-protein
transcript-2/leukocyte	NN	O	I-protein
Ig-like	NN	O	I-protein
receptor	NN	O	I-protein
1	NN	O	I-protein
on	NN	O	O
NKL	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
HLA-B27	NN	O	B-protein
on	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
leads	NN	O	O
to	NN	O	O
an	NN	O	O
impairment	NN	O	O
of	NN	O	O
NKL	NN	O	O
natural	NN	O	O
cytotoxicity	NN	O	O
,	NN	O	O
which	NN	O	O
correlates	NN	O	O
with	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
JunB	NN	O	O
,	NN	O	O
FosB	NN	O	O
,	NN	O	O
and	NN	O	O
c-Fos	NN	O	O
transcription	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
their	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
thus	NN	O	O
indicate	NN	O	O
that	NN	O	O
,	NN	O	O
despite	NN	O	O
the	NN	O	O
rapidity	NN	O	O
of	NN	O	O
NK	NN	O	O
cell-mediated	NN	O	O
lysis	NN	O	O
,	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
activated	NN	O	O
during	NN	O	O
the	NN	O	O
early	NN	O	O
stage	NN	O	O
of	NN	O	O
NK	NN	O	O
cell	NN	O	O
cytolytic	NN	O	O
programs	NN	O	O
and	NN	O	O
that	NN	O	O
engagement	NN	O	O
of	NN	O	O
NK	NN	O	B-protein
cell	NN	O	I-protein
inhibitory	NN	O	I-protein
receptors	NN	O	I-protein
for	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
molecules	NN	O	I-protein
impairs	NN	O	O
the	NN	O	O
very	NN	O	O
early	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Evidence	NN	O	O
for	NN	O	O
a	NN	O	O
polyclonal	NN	O	O
etiology	NN	O	O
of	NN	O	O
palmar	NN	O	O
fibromatosis	NN	O	O
.	NN	O	O

X	NN	O	B-DNA
chromosome	NN	O	I-DNA
inactivation	NN	O	O
patterns	NN	O	O
at	NN	O	O
the	NN	O	O
androgen	NN	O	O
receptor	NN	O	O
locus	NN	O	O
were	NN	O	O
evaluated	NN	O	O
to	NN	O	O
determine	NN	O	O
clonality	NN	O	O
in	NN	O	O
microdissected	NN	O	O
lesional	NN	O	O
tissue	NN	O	O
and	NN	O	O
in	NN	O	O
leukocytes	NN	O	B-cell_type
from	NN	O	O
2	NN	O	O
women	NN	O	O
with	NN	O	O
Dupuytren	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
tissue	NN	O	O
from	NN	O	O
both	NN	O	O
patients	NN	O	O
generated	NN	O	O
a	NN	O	O
polyclonal	NN	O	O
pattern	NN	O	O
of	NN	O	O
X	NN	O	B-DNA
chromosome	NN	O	I-DNA
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
androgen	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
finding	NN	O	O
supports	NN	O	O
a	NN	O	O
polyclonal	NN	O	O
reactive	NN	O	O
process	NN	O	O
as	NN	O	O
the	NN	O	O
underlying	NN	O	O
etiology	NN	O	O
for	NN	O	O
palmar	NN	O	O
fibromatosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
cyclooxygenase-2	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
4-trifluoromethyl	NN	O	O
derivatives	NN	O	O
of	NN	O	O
salicylate	NN	O	O
,	NN	O	O
triflusal	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
deacetylated	NN	O	O
metabolite	NN	O	O
,	NN	O	O
2-hydroxy-4-trifluoromethylbenzoic	NN	O	O
acid	NN	O	O
.	NN	O	O

The	NN	O	O
therapeutic	NN	O	O
potential	NN	O	O
of	NN	O	O
drugs	NN	O	O
that	NN	O	O
block	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
cyclooxygenase-2	NN	O	B-protein
has	NN	O	O
been	NN	O	O
emphasized	NN	O	O
.	NN	O	O

When	NN	O	O
two	NN	O	O
4-trifluoromethyl	NN	O	O
salicylate	NN	O	O
derivatives	NN	O	O
[	NN	O	O
2-acetoxy-4-trifluoromethyl-benzoic	NN	O	O
acid	NN	O	O
(	NN	O	O
triflusal	NN	O	O
)	NN	O	O
and	NN	O	O
its	NN	O	O
deacetylated	NN	O	O
metabolite	NN	O	O
2-hydroxy-4-trifluoromethylbenzoic	NN	O	O
acid	NN	O	O
(	NN	O	O
HTB	NN	O	O
)	NN	O	O
]	NN	O	O
were	NN	O	O
compared	NN	O	O
with	NN	O	O
aspirin	NN	O	O
and	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
as	NN	O	O
cyclooxygenase-2	NN	O	B-protein
(	NN	O	O
COX-2	NN	O	O
)	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
in	NN	O	O
bacterial	NN	O	O
lipopolysaccharide-activated	NN	O	O
human	NN	O	O
blood	NN	O	O
,	NN	O	O
triflusal	NN	O	O
,	NN	O	O
aspirin	NN	O	O
,	NN	O	O
and	NN	O	O
HTB	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
,	NN	O	O
inhibited	NN	O	O
COX-2-mediated	NN	O	O
prostaglandin	NN	O	O
E2	NN	O	O
(	NN	O	O
PGE2	NN	O	O
)	NN	O	O
production	NN	O	O
(	NN	O	O
IC50	NN	O	O
=	NN	O	O
0.16	NN	O	O
,	NN	O	O
0.18	NN	O	O
,	NN	O	O
0.39	NN	O	O
,	NN	O	O
and	NN	O	O
>	NN	O	O
10	NN	O	O
mM	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
only	NN	O	O
triflusal	NN	O	O
and	NN	O	O
aspirin	NN	O	O
inhibited	NN	O	O
purified	NN	O	B-protein
COX-2	NN	O	I-protein
enzyme	NN	O	I-protein
.	NN	O	O

To	NN	O	O
test	NN	O	O
this	NN	O	O
apparent	NN	O	O
discrepancy	NN	O	O
,	NN	O	O
we	NN	O	O
realized	NN	O	O
that	NN	O	O
HTB	NN	O	O
and	NN	O	O
triflusal	NN	O	O
(	NN	O	O
but	NN	O	O
neither	NN	O	O
aspirin	NN	O	O
nor	NN	O	O
salicylate	NN	O	O
)	NN	O	O
produced	NN	O	O
a	NN	O	O
concentration-dependent	NN	O	O
inhibition	NN	O	O
of	NN	O	O
COX-2	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
human	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
observation	NN	O	O
was	NN	O	O
further	NN	O	O
confirmed	NN	O	O
in	NN	O	O
a	NN	O	O
rat	NN	O	O
air	NN	O	O
pouch	NN	O	O
model	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
both	NN	O	O
aspirin	NN	O	O
and	NN	O	O
triflusal	NN	O	O
inhibited	NN	O	O
PGE2	NN	O	O
production	NN	O	O
(	NN	O	O
ID50	NN	O	O
=	NN	O	O
18.9	NN	O	O
and	NN	O	O
11.4	NN	O	O
mg/kg	NN	O	O
p.o.	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
but	NN	O	O
only	NN	O	O
triflusal-treated	NN	O	O
animals	NN	O	O
showed	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
COX-2	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

This	NN	O	O
different	NN	O	O
behavior	NN	O	O
may	NN	O	O
be	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
HTB	NN	O	O
and	NN	O	O
triflusal	NN	O	O
to	NN	O	O
block	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
to	NN	O	O
a	NN	O	O
higher	NN	O	O
extent	NN	O	O
than	NN	O	O
aspirin	NN	O	O
and	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
COX-2	NN	O	O
activity	NN	O	O
at	NN	O	O
therapeutic	NN	O	O
concentrations	NN	O	O
,	NN	O	O
triflusal	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
block	NN	O	O
through	NN	O	O
its	NN	O	O
metabolite	NN	O	O
HTB	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
new	NN	O	O
enzyme	NN	O	O
,	NN	O	O
and	NN	O	O
hence	NN	O	O
the	NN	O	O
resumption	NN	O	O
of	NN	O	O
PGE2	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Triflusal	NN	O	O
and	NN	O	O
HTB	NN	O	O
may	NN	O	O
exert	NN	O	O
beneficial	NN	O	O
effects	NN	O	O
in	NN	O	O
processes	NN	O	O
in	NN	O	O
which	NN	O	O
de	NN	O	O
novo	NN	O	O
COX-2	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
involved	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
broader	NN	O	O
sense	NN	O	O
,	NN	O	O
in	NN	O	O
pathological	NN	O	O
situations	NN	O	O
in	NN	O	O
which	NN	O	O
genes	NN	O	O
under	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
control	NN	O	O
are	NN	O	O
up-regulated	NN	O	O
.	NN	O	O

-DOCSTART-	O

Evidence	NN	O	O
for	NN	O	O
distinct	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
CD34+	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
to	NN	O	O
dendritic	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
from	NN	O	O
a	NN	O	O
human	NN	O	O
cell	NN	O	O
line	NN	O	O
model	NN	O	O
.	NN	O	O

Intracellular	NN	O	O
signals	NN	O	O
that	NN	O	O
mediate	NN	O	O
differentiation	NN	O	O
of	NN	O	O
pluripotent	NN	O	B-cell_type
hemopoietic	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
to	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
DC	NN	O	B-cell_type
)	NN	O	O
are	NN	O	O
largely	NN	O	O
undefined	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
(	NN	O	O
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
alone	NN	O	O
)	NN	O	O
specifically	NN	O	O
induces	NN	O	O
differentiation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_line
human	NN	O	I-cell_line
CD34+	NN	O	I-cell_line
hemopoietic	NN	O	I-cell_line
progenitor	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
HPC	NN	O	B-cell_line
)	NN	O	O
to	NN	O	O
mature	NN	O	B-cell_type
DC	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
now	NN	O	O
find	NN	O	O
that	NN	O	O
cytokine-driven	NN	O	O
(	NN	O	O
granulocyte-macrophage	NN	O	O
CSF	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	O
)	NN	O	O
CD34+	NN	O	O
HPC	NN	O	B-cell_line
--	NN	O	O
>	NN	O	O
DC	NN	O	O
differentiation	NN	O	O
is	NN	O	O
preferentially	NN	O	O
blocked	NN	O	O
by	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
identify	NN	O	O
intracellular	NN	O	O
signals	NN	O	O
and	NN	O	O
downstream	NN	O	O
events	NN	O	O
important	NN	O	O
in	NN	O	O
CD34+	NN	O	B-cell_line
HPC	NN	O	I-cell_line
--	NN	O	O
>	NN	O	O
DC	NN	O	B-cell_type
differentiation	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
a	NN	O	O
human	NN	O	O
leukemic	NN	O	O
cell	NN	O	O
line	NN	O	O
model	NN	O	O
of	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

The	NN	O	O
CD34+	NN	O	B-cell_line
myelomonocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
KG1	NN	O	B-cell_line
differentiates	NN	O	O
into	NN	O	O
dendritic-like	NN	O	O
cells	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
granulocyte-macrophage	NN	O	B-protein
CSF	NN	O	I-protein
plus	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
or	NN	O	O
PMA	NN	O	O
(	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
the	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
ionomycin	NN	O	O
,	NN	O	O
or	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
with	NN	O	O
different	NN	O	O
stimuli	NN	O	O
mediating	NN	O	O
different	NN	O	O
aspects	NN	O	O
of	NN	O	O
the	NN	O	O
process	NN	O	O
.	NN	O	O

Phenotypic	NN	O	O
DC	NN	O	B-cell_type
characteristics	NN	O	O
of	NN	O	O
KG1	NN	O	B-cell_line
dendritic-like	NN	O	I-cell_line
cells	NN	O	I-cell_line
include	NN	O	O
morphology	NN	O	O
(	NN	O	O
loosely	NN	O	B-cell_type
adherent	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
long	NN	O	O
neurite	NN	O	O
processes	NN	O	O
)	NN	O	O
,	NN	O	O
MHC	NN	O	B-protein
I+/	NN	O	O
MHC	NN	O	B-protein
IIbright/CD83+/CD86+/	NN	O	O
CD14	NN	O	B-protein
-	NN	O	O
surface	NN	O	O
Ag	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
RelB	NN	O	B-protein
and	NN	O	O
DC	NN	O	B-cell_type
-CK1	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Functional	NN	O	O
DC	NN	O	B-cell_type
characteristics	NN	O	O
include	NN	O	O
fluid	NN	O	O
phase	NN	O	O
macromolecule	NN	O	O
uptake	NN	O	O
(	NN	O	O
FITC-dextran	NN	O	O
)	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
KG1	NN	O	B-cell_line
to	NN	O	O
the	NN	O	O
PMA-unresponsive	NN	O	B-cell_line
subline	NN	O	I-cell_line
KG1a	NN	O	I-cell_line
reveals	NN	O	O
differences	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
receptors	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	I-protein
2	NN	O	I-protein
;	NN	O	O
PKC	NN	O	B-protein
isoforms	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	I-protein
beta	NN	O	I-protein
I	NN	O	I-protein
,	NN	O	I-protein
beta	NN	O	I-protein
II	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
mu	NN	O	I-protein
;	NN	O	O
and	NN	O	O
RelB	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
components/pathways	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
DC	NN	O	B-cell_type
differentiation	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
cytokine	NN	O	O
or	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
stimulation	NN	O	O
of	NN	O	O
KG1	NN	O	B-cell_line
is	NN	O	O
a	NN	O	O
model	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
CD34+	NN	O	I-cell_line
HPC	NN	O	I-cell_line
to	NN	O	O
DC	NN	O	B-cell_type
differentiation	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
specific	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
mediate	NN	O	O
specific	NN	O	O
events	NN	O	O
in	NN	O	O
DC	NN	O	B-cell_type
lineage	NN	O	O
commitment	NN	O	O
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
adenylate	NN	O	B-protein
cyclase	NN	O	I-protein
and	NN	O	O
p70	NN	O	B-protein
(	NN	O	I-protein
S6	NN	O	I-protein
)	NN	O	I-protein
-kinase	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
IL-10	NN	O	B-protein
up-regulation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
by	NN	O	O
gp41	NN	O	B-protein
envelope	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
recombinant	NN	O	B-protein
gp41	NN	O	I-protein
(	NN	O	O
aa565-647	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
HIV-1	NN	O	B-protein
transmembrane	NN	O	I-protein
glycoprotein	NN	O	I-protein
,	NN	O	O
stimulates	NN	O	O
interleukin-10	NN	O	B-protein
(	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
production	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
signal	NN	O	O
cascade	NN	O	O
transducing	NN	O	O
this	NN	O	O
effect	NN	O	O
is	NN	O	O
not	NN	O	O
yet	NN	O	O
clear	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
whether	NN	O	O
gp41	NN	O	B-protein
-induced	NN	O	O
IL-10	NN	O	B-protein
up-regulation	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
previously	NN	O	O
described	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
cAMP	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
pathways	NN	O	O
.	NN	O	O

gp41	NN	O	B-protein
induced	NN	O	O
cAMP	NN	O	O
accumulation	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
in	NN	O	O
a	NN	O	O
time-	NN	O	O
and	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
and	NN	O	O
the	NN	O	O
adenylate	NN	O	B-protein
cyclase	NN	O	I-protein
inhibitor	NN	O	O
SQ	NN	O	O
22536	NN	O	O
suppressed	NN	O	O
gp41	NN	O	B-protein
-induced	NN	O	O
IL-10	NN	O	B-protein
production	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
gp41	NN	O	B-protein
failed	NN	O	O
to	NN	O	O
stimulate	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
as	NN	O	O
much	NN	O	O
as	NN	O	O
no	NN	O	O
NF-kappaB	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
main	NN	O	B-DNA
NF-kappaB-binding	NN	O	I-DNA
site	NN	O	I-DNA
2	NN	O	O
of	NN	O	O
the	NN	O	O
IL-10	NN	O	B-DNA
promoter	NN	O	I-DNA
after	NN	O	O
addition	NN	O	O
of	NN	O	O
gp41	NN	O	B-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
examined	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
other	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
.	NN	O	O

Specific	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
p70	NN	O	B-protein
(	NN	O	I-protein
S6	NN	O	I-protein
)	NN	O	I-protein
-kinase	NN	O	I-protein
(	NN	O	O
rapamycin	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
Gi	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
pertussis	NN	O	B-protein
toxin	NN	O	I-protein
)	NN	O	O
,	NN	O	O
prevented	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
production	NN	O	O
by	NN	O	O
gp41	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
while	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
the	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
(	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
)	NN	O	O
(	NN	O	O
wortmannin	NN	O	O
)	NN	O	O
and	NN	O	O
mitogen-activated	NN	O	O
protein	NN	O	O
kinase	NN	O	O
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
pathway	NN	O	O
(	NN	O	O
PD	NN	O	O
98059	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

Thus	NN	O	O
HIV-1	NN	O	O
gp41	NN	O	B-protein
-induced	NN	O	O
IL-10	NN	O	B-protein
up-regulation	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
may	NN	O	O
not	NN	O	O
involve	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
MAPK	NN	O	B-protein
,	NN	O	O
or	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
but	NN	O	O
rather	NN	O	O
may	NN	O	O
operate	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
adenylate	NN	O	B-protein
cyclase	NN	O	I-protein
and	NN	O	O
pertussis-toxin-sensitive	NN	O	B-protein
Gi/Go	NN	O	I-protein
protein	NN	O	I-protein
to	NN	O	O
effect	NN	O	O
p70	NN	O	B-protein
(	NN	O	I-protein
S6	NN	O	I-protein
)	NN	O	I-protein
-kinase	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
myelopoiesis	NN	O	O
and	NN	O	O
leukemogenesis	NN	O	O
.	NN	O	O

Acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
(	NN	O	O
AML	NN	O	O
)	NN	O	O
is	NN	O	O
still	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
mortality	NN	O	O
of	NN	O	O
60	NN	O	O
to	NN	O	O
80	NN	O	O
%	NN	O	O
.	NN	O	O

AML	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
block	NN	O	O
in	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
PU.1	NN	O	B-protein
and	NN	O	O
C/EBPalpha	NN	O	B-protein
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
normal	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
from	NN	O	O
stem	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
monocytes	NN	O	B-cell_type
or	NN	O	O
granulocytes	NN	O	O
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
PU.1	NN	O	B-protein
induces	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
M-CSF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
whereas	NN	O	O
C/EBPalpha	NN	O	B-protein
increases	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
G-CSF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
leads	NN	O	O
to	NN	O	O
mature	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
AML	NN	O	O
,	NN	O	O
chromosomal	NN	O	O
aberrations	NN	O	O
result	NN	O	O
in	NN	O	O
oncoproteins	NN	O	B-protein
such	NN	O	O
as	NN	O	O
AML1/ETO	NN	O	B-protein
,	NN	O	O
PML/RARalpha	NN	O	B-protein
,	NN	O	O
or	NN	O	O
activated	NN	O	B-protein
Ras	NN	O	I-protein
,	NN	O	O
which	NN	O	O
can	NN	O	O
deregulate	NN	O	O
genes	NN	O	O
important	NN	O	O
for	NN	O	O
normal	NN	O	O
myelopoiesis	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
AML1/ETO	NN	O	B-protein
can	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
C/EBPalpha	NN	O	B-protein
,	NN	O	O
inhibit	NN	O	O
C/EBPalpha	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
block	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
AML1/ETO	NN	O	B-protein
can	NN	O	O
also	NN	O	O
synergize	NN	O	O
with	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AML1	NN	O	B-protein
to	NN	O	O
enhance	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-DNA
receptor	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
the	NN	O	O
PML/RARalpha	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
causes	NN	O	O
transcriptional	NN	O	O
repression	NN	O	O
by	NN	O	O
recruiting	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
corepressor	NN	O	I-protein
(	NN	O	I-protein
N-CoR	NN	O	I-protein
)	NN	O	I-protein
histone	NN	O	I-protein
deacetylase	NN	O	I-protein
complex	NN	O	I-protein
to	NN	O	O
the	NN	O	O
DNA	NN	O	O
,	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
decreased	NN	O	O
histone	NN	O	B-protein
acetylation	NN	O	O
and	NN	O	O
a	NN	O	O
repressive	NN	O	O
chromatin	NN	O	B-DNA
organization	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
methods	NN	O	O
to	NN	O	O
investigate	NN	O	O
whether	NN	O	O
and	NN	O	O
how	NN	O	O
signaling	NN	O	O
agonists	NN	O	O
induce	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
and	NN	O	O
how	NN	O	O
oncoproteins	NN	O	B-protein
might	NN	O	O
cause	NN	O	O
AML	NN	O	O
by	NN	O	O
modulating	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
pivotal	NN	O	O
for	NN	O	O
normal	NN	O	O
myeloid	NN	O	O
development	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-ATc	NN	O	B-protein
isoforms	NN	O	I-protein
are	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
and	NN	O	O
regulated	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
mast	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NF	NN	O	B-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
denotes	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
the	NN	O	O
activation-dependent	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
immunologically	NN	O	B-protein
important	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

At	NN	O	O
least	NN	O	O
four	NN	O	O
distinct	NN	O	O
genes	NN	O	O
encode	NN	O	O
the	NN	O	O
various	NN	O	O
family	NN	O	O
members	NN	O	O
,	NN	O	O
and	NN	O	O
several	NN	O	O
isoforms	NN	O	O
of	NN	O	O
these	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
as	NN	O	O
well	NN	O	O
.	NN	O	O

The	NN	O	O
overlapping	NN	O	O
expression	NN	O	O
patterns	NN	O	O
and	NN	O	O
similar	NN	O	O
in	NN	O	O
vitro	NN	O	O
binding	NN	O	O
and	NN	O	O
trans-activation	NN	O	O
activities	NN	O	O
on	NN	O	O
various	NN	O	O
promoter	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
NF-AT-regulated	NN	O	B-DNA
genes	NN	O	I-DNA
suggest	NN	O	O
some	NN	O	O
redundancy	NN	O	O
in	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
analysis	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
-deficient	NN	O	O
mice	NN	O	O
supports	NN	O	O
the	NN	O	O
idea	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
tissue-	NN	O	O
and	NN	O	O
gene-	NN	O	O
specific	NN	O	O
functions	NN	O	O
as	NN	O	O
well	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-DNA
cDNAs	NN	O	I-DNA
in	NN	O	O
murine	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
majority	NN	O	O
of	NN	O	O
clones	NN	O	O
identified	NN	O	O
correspond	NN	O	O
to	NN	O	O
two	NN	O	O
NF-ATc	NN	O	B-protein
isoforms	NN	O	I-protein
that	NN	O	O
differ	NN	O	O
only	NN	O	O
in	NN	O	O
their	NN	O	O
amino-terminal	NN	O	B-protein
sequence	NN	O	I-protein
.	NN	O	O

Despite	NN	O	O
minimal	NN	O	O
discrepancies	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	B-DNA
region	NN	O	I-DNA
,	NN	O	O
there	NN	O	O
are	NN	O	O
striking	NN	O	O
tissue-	NN	O	O
and	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
differences	NN	O	O
in	NN	O	O
isoform	NN	O	O
expression	NN	O	O
patterns	NN	O	O
.	NN	O	O

Detection	NN	O	O
of	NN	O	O
NF-ATc.alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
strictly	NN	O	O
dependent	NN	O	O
on	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
in	NN	O	O
both	NN	O	O
T	NN	O	B-cell_line
and	NN	O	I-cell_line
mast	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
beta	NN	O	B-protein
isoform	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
very	NN	O	O
low	NN	O	O
constitutive	NN	O	O
levels	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
but	NN	O	O
is	NN	O	O
only	NN	O	O
up-regulated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
mast	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
delivered	NN	O	O
through	NN	O	O
the	NN	O	O
FcepsilonRI	NN	O	B-protein
or	NN	O	O
via	NN	O	O
calcium	NN	O	O
ionophores	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
another	NN	O	O
level	NN	O	O
of	NN	O	O
regulation	NN	O	O
within	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
family	NN	O	I-protein
that	NN	O	O
can	NN	O	O
contribute	NN	O	O
to	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Crossreactive	NN	O	O
recognition	NN	O	O
of	NN	O	O
viral	NN	O	O
,	NN	O	O
self	NN	O	O
,	NN	O	O
and	NN	O	O
bacterial	NN	O	O
peptide	NN	O	O
ligands	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
class	NN	O	I-cell_type
I-restricted	NN	O	I-cell_type
cytotoxic	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
clonotypes	NN	O	I-cell_type
:	NN	O	I-cell_type
implications	NN	O	O
for	NN	O	O
molecular	NN	O	O
mimicry	NN	O	O
in	NN	O	O
autoimmune	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
immunodominant	NN	O	O
,	NN	O	O
CD8	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cytotoxic	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
(	NN	O	O
CTL	NN	O	O
)	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
HLA-B8-restricted	NN	O	O
peptide	NN	O	O
,	NN	O	O
RAKFKQLL	NN	O	O
,	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
immediate-early	NN	O	I-protein
antigen	NN	O	I-protein
,	NN	O	O
BZLF1	NN	O	B-protein
,	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
diverse	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
TCR	NN	O	I-protein
)	NN	O	I-protein
repertoire	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
diversity	NN	O	O
can	NN	O	O
be	NN	O	O
partitioned	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
crossreactive	NN	O	O
cytotoxicity	NN	O	O
patterns	NN	O	O
involving	NN	O	O
the	NN	O	O
recognition	NN	O	O
of	NN	O	O
a	NN	O	O
self	NN	O	O
peptide-RSKFRQIV-located	NN	O	O
in	NN	O	O
a	NN	O	O
serine/threonine	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
a	NN	O	O
bacterial	NN	O	O
peptide-RRKYKQII-located	NN	O	O
in	NN	O	O
Staphylococcus	NN	O	B-protein
aureus	NN	O	I-protein
replication	NN	O	I-protein
initiation	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
CTL	NN	O	B-cell_line
clones	NN	O	I-cell_line
that	NN	O	O
recognized	NN	O	O
the	NN	O	O
viral	NN	O	O
,	NN	O	O
self	NN	O	O
,	NN	O	O
and	NN	O	O
bacterial	NN	O	O
peptides	NN	O	O
expressed	NN	O	O
a	NN	O	O
highly	NN	O	O
restricted	NN	O	O
alphabeta	NN	O	O
TCR	NN	O	B-protein
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
CTL	NN	O	B-cell_line
clones	NN	O	I-cell_line
that	NN	O	O
recognized	NN	O	O
viral	NN	O	O
and	NN	O	O
self	NN	O	O
peptides	NN	O	O
were	NN	O	O
more	NN	O	O
oligoclonal	NN	O	O
,	NN	O	O
whereas	NN	O	O
clones	NN	O	O
that	NN	O	O
strictly	NN	O	O
recognized	NN	O	O
the	NN	O	O
viral	NN	O	O
peptide	NN	O	O
displayed	NN	O	O
a	NN	O	O
diverse	NN	O	O
TCR	NN	O	B-protein
profile	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
self	NN	O	O
and	NN	O	O
bacterial	NN	O	O
peptides	NN	O	O
equally	NN	O	O
were	NN	O	O
substantially	NN	O	O
less	NN	O	O
effective	NN	O	O
than	NN	O	O
the	NN	O	O
cognate	NN	O	O
viral	NN	O	O
peptide	NN	O	O
in	NN	O	O
sensitizing	NN	O	O
target	NN	O	O
cell	NN	O	O
lysis	NN	O	O
,	NN	O	O
and	NN	O	O
also	NN	O	O
resulted	NN	O	O
only	NN	O	O
in	NN	O	O
a	NN	O	O
weak	NN	O	O
reactivation	NN	O	O
of	NN	O	O
memory	NN	O	B-cell_type
CTLs	NN	O	I-cell_type
in	NN	O	O
limiting	NN	O	O
dilution	NN	O	O
assays	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
cognate	NN	O	O
peptide	NN	O	O
was	NN	O	O
highly	NN	O	O
immunogenic	NN	O	O
.	NN	O	O

The	NN	O	O
described	NN	O	O
crossreactions	NN	O	O
show	NN	O	O
that	NN	O	O
human	NN	O	O
antiviral	NN	O	O
,	NN	O	O
CD8	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
CTL	NN	O	O
responses	NN	O	O
can	NN	O	O
be	NN	O	O
shaped	NN	O	O
by	NN	O	O
peptide	NN	O	O
ligands	NN	O	O
derived	NN	O	O
from	NN	O	O
autoantigens	NN	O	O
and	NN	O	O
environmental	NN	O	O
bacterial	NN	O	O
antigens	NN	O	O
,	NN	O	O
thereby	NN	O	O
providing	NN	O	O
a	NN	O	O
firm	NN	O	O
structural	NN	O	O
basis	NN	O	O
for	NN	O	O
molecular	NN	O	O
mimicry	NN	O	O
involving	NN	O	O
class	NN	O	O
I-restricted	NN	O	B-cell_line
CTLs	NN	O	I-cell_line
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
autoimmune	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

RFLAT-1	NN	O	B-protein
:	NN	O	O
a	NN	O	O
new	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
activates	NN	O	O
RANTES	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

RANTES	NN	O	B-protein
(	NN	O	O
Regulated	NN	O	B-protein
upon	NN	O	I-protein
Activation	NN	O	I-protein
,	NN	O	I-protein
Normal	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
Expressed	NN	O	I-protein
and	NN	O	I-protein
Secreted	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
chemoattractant	NN	O	B-protein
cytokine	NN	O	I-protein
(	NN	O	O
chemokine	NN	O	B-protein
)	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
infiltrate	NN	O	O
and	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
entry	NN	O	O
into	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

RANTES	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
late	NN	O	O
(	NN	O	O
3-5	NN	O	O
days	NN	O	O
)	NN	O	O
after	NN	O	O
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
expression	NN	O	O
cloning	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
the	NN	O	O
first	NN	O	O
``	NN	O	O
late	NN	O	O
''	NN	O	O
T	NN	O	B-protein
lymphocyte	NN	O	I-protein
associated	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
named	NN	O	O
it	NN	O	O
``	NN	O	O
RANTES	NN	O	B-protein
Factor	NN	O	I-protein
of	NN	O	I-protein
Late	NN	O	I-protein
Activated	NN	O	I-protein
T	NN	O	I-protein
Lymphocytes-1	NN	O	I-protein
``	NN	O	O
(	NN	O	O
RFLAT-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

RFLAT-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
novel	NN	O	O
,	NN	O	O
phosphorylated	NN	O	O
,	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
3	NN	O	O
days	NN	O	O
after	NN	O	O
activation	NN	O	O
,	NN	O	O
coincident	NN	O	O
with	NN	O	O
RANTES	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

While	NN	O	O
Rel	NN	O	B-protein
proteins	NN	O	I-protein
play	NN	O	O
the	NN	O	O
dominant	NN	O	O
role	NN	O	O
in	NN	O	O
RANTES	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
fibroblasts	NN	O	O
,	NN	O	O
RFLAT-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
strong	NN	O	O
transactivator	NN	O	O
for	NN	O	O
RANTES	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Fas	NN	O	B-protein
ligand	NN	O	I-protein
induction	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
redox	NN	O	O
through	NN	O	O
a	NN	O	O
calcineurin-nuclear	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
activated	NN	O	O
T	NN	O	O
cell-dependent	NN	O	O
pathway	NN	O	O
.	NN	O	O

Fas	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
FasL	NN	O	B-protein
)	NN	O	O
on	NN	O	O
cytotoxic	NN	O	O
lymphocytes	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
mediating	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
activated	NN	O	O
lymphocytes	NN	O	O
and	NN	O	O
other	NN	O	O
target	NN	O	O
cells	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
reported	NN	O	O
that	NN	O	O
NK	NN	O	O
cell	NN	O	O
functions	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
proliferation	NN	O	O
,	NN	O	O
cell	NN	O	O
death	NN	O	O
,	NN	O	O
and	NN	O	O
killing	NN	O	O
activity	NN	O	O
,	NN	O	O
are	NN	O	O
subject	NN	O	O
to	NN	O	O
regulation	NN	O	O
by	NN	O	O
cellular	NN	O	O
redox	NN	O	O
status	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
mRNA	NN	O	B-RNA
in	NN	O	O
activated	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
also	NN	O	O
regulated	NN	O	O
by	NN	O	O
redox	NN	O	O
.	NN	O	O

Ligation	NN	O	O
of	NN	O	O
CD16	NN	O	B-protein
on	NN	O	O
IL-2-preactivated	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
resulted	NN	O	O
in	NN	O	O
reduction	NN	O	O
of	NN	O	O
intracellular	NN	O	O
peroxide	NN	O	O
level	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
induction	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

This	NN	O	O
CD16	NN	O	B-protein
-induced	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
suppressed	NN	O	O
by	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
,	NN	O	O
including	NN	O	O
thiol	NN	O	O
deprivation	NN	O	O
or	NN	O	O
treatment	NN	O	O
with	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
(	NN	O	O
H2O2	NN	O	O
)	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
thiol-reducing	NN	O	O
compounds	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
L-cystine	NN	O	O
,	NN	O	O
2-ME	NN	O	O
,	NN	O	O
or	NN	O	O
N-acetyl	NN	O	O
cysteine	NN	O	O
,	NN	O	O
restored	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
CD16	NN	O	B-protein
stimulation	NN	O	O
requires	NN	O	O
cellular	NN	O	O
reducing	NN	O	O
status	NN	O	O
for	NN	O	O
FasL	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Because	NN	O	O
FasL	NN	O	O
gene	NN	O	O
activation	NN	O	O
following	NN	O	O
CD16	NN	O	B-protein
cross-linking	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
oxidative	NN	O	O
stresses	NN	O	O
on	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
revealed	NN	O	O
that	NN	O	O
both	NN	O	O
thiol	NN	O	O
insufficiency	NN	O	O
and	NN	O	O
H2O2	NN	O	O
treatment	NN	O	O
suppressed	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
and	NN	O	O
that	NN	O	O
addition	NN	O	O
of	NN	O	O
thiol-reducing	NN	O	O
compounds	NN	O	O
reversed	NN	O	O
or	NN	O	O
even	NN	O	O
enhanced	NN	O	O
it	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
these	NN	O	O
oxidative	NN	O	O
stresses	NN	O	O
inhibited	NN	O	O
activity	NN	O	O
of	NN	O	O
calcineurin	NN	O	O
,	NN	O	O
a	NN	O	O
serine/	NN	O	O
threonine	NN	O	B-protein
phosphatase	NN	O	I-protein
that	NN	O	O
regulates	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
suppression	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
and	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
a	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
inhibits	NN	O	O
FasL	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
further	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
thiol-reducing	NN	O	O
compounds	NN	O	O
might	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
maintenance	NN	O	O
of	NN	O	O
optimal	NN	O	O
NK	NN	O	O
functions	NN	O	O
under	NN	O	O
physiologic	NN	O	O
oxidative	NN	O	O
conditions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Heterogeneity	NN	O	O
of	NN	O	O
clonal	NN	O	O
development	NN	O	O
in	NN	O	O
chronic	NN	O	B-DNA
myeloproliferative	NN	O	I-DNA
disorders	NN	O	I-DNA
.	NN	O	O

Recent	NN	O	O
reports	NN	O	O
have	NN	O	O
suggested	NN	O	O
a	NN	O	O
previously	NN	O	O
unexpected	NN	O	O
variability	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
dominant	NN	O	B-cell_line
neoplastic	NN	O	I-cell_line
clone	NN	O	I-cell_line
in	NN	O	O
myeloproliferative	NN	O	O
disorders	NN	O	O
(	NN	O	O
MPD	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
evaluated	NN	O	O
49	NN	O	O
female	NN	O	O
patients	NN	O	O
with	NN	O	O
MPD	NN	O	O
and	NN	O	O
informative	NN	O	O
at	NN	O	O
the	NN	O	O
X-linked	NN	O	B-DNA
androgen	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
AR	NN	O	I-DNA
)	NN	O	I-DNA
locus	NN	O	I-DNA
to	NN	O	O
establish	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
chromosome	NN	O	I-DNA
inactivation	NN	O	O
pattern	NN	O	O
of	NN	O	O
hemopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Whereas	NN	O	O
in	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CML	NN	O	O
)	NN	O	O
the	NN	O	O
granulocytes	NN	O	B-cell_type
(	NN	O	O
PMN	NN	O	B-cell_type
)	NN	O	O
were	NN	O	O
uniformly	NN	O	O
of	NN	O	O
monoclonal	NN	O	O
origin	NN	O	O
,	NN	O	O
a	NN	O	O
striking	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
clonal	NN	O	O
development	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
PMN	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
other	NN	O	O
MPD	NN	O	O
,	NN	O	O
with	NN	O	O
up	NN	O	O
to	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
them	NN	O	O
expressing	NN	O	O
a	NN	O	O
polyclonal	NN	O	O
pattern	NN	O	O
of	NN	O	O
X	NN	O	O
inactivation	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD80	NN	O	B-protein
and	NN	O	O
CD86	NN	O	B-protein
are	NN	O	O
not	NN	O	O
equivalent	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

Ligation	NN	O	O
of	NN	O	O
either	NN	O	O
CD80	NN	O	B-protein
(	NN	O	O
B7-1	NN	O	B-protein
)	NN	O	O
or	NN	O	O
CD86	NN	O	B-protein
(	NN	O	O
B7-2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
two	NN	O	O
principal	NN	O	O
ligands	NN	O	O
for	NN	O	O
CD28	NN	O	B-protein
,	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
skew	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
toward	NN	O	O
Th1	NN	O	O
or	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
early	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
recruited	NN	O	O
following	NN	O	O
T	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
with	NN	O	O
either	NN	O	O
CD80	NN	O	B-protein
or	NN	O	O
CD86	NN	O	B-protein
.	NN	O	O

Purified	NN	O	B-cell_line
human	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
Chinese	NN	O	O
hamster	NN	O	O
ovary	NN	O	O
(	NN	O	O
CHO	NN	O	O
)	NN	O	O
cells	NN	O	O
expressing	NN	O	O
either	NN	O	O
human	NN	O	B-protein
CD80	NN	O	I-protein
(	NN	O	O
CHO-CD80	NN	O	B-protein
)	NN	O	O
or	NN	O	O
human	NN	O	B-protein
CD86	NN	O	I-protein
(	NN	O	O
CHO-CD86	NN	O	B-protein
)	NN	O	O
or	NN	O	O
with	NN	O	O
anti-CD28	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
mAb	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
,	NN	O	O
both	NN	O	O
CHO-CD80	NN	O	B-protein
and	NN	O	O
CHO-CD86	NN	O	B-protein
,	NN	O	O
like	NN	O	O
anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
,	NN	O	O
were	NN	O	O
capable	NN	O	O
of	NN	O	O
stimulating	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
from	NN	O	O
both	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Both	NN	O	O
CHO-CD80	NN	O	B-protein
and	NN	O	O
CHO-CD86	NN	O	B-protein
,	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
,	NN	O	O
costimulated	NN	O	O
NFAT	NN	O	B-protein
-dependent	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Several	NN	O	O
intracellular	NN	O	O
signaling	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
CBL	NN	O	B-protein
and	NN	O	O
VAV	NN	O	B-protein
,	NN	O	O
were	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
CD80	NN	O	B-protein
,	NN	O	O
CD86	NN	O	B-protein
,	NN	O	O
and	NN	O	O
anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
although	NN	O	O
stimulation	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
either	NN	O	O
CHO-CD80	NN	O	B-protein
or	NN	O	O
anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
resulted	NN	O	O
in	NN	O	O
robust	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
itself	NN	O	O
,	NN	O	O
ligation	NN	O	O
with	NN	O	O
CHO-CD86	NN	O	B-protein
was	NN	O	O
unable	NN	O	O
to	NN	O	O
induce	NN	O	O
detectable	NN	O	O
CD28	NN	O	B-protein
tyrosyl	NN	O	O
phosphorylation	NN	O	O
over	NN	O	O
a	NN	O	O
range	NN	O	O
of	NN	O	O
stimulation	NN	O	O
conditions	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
phosphoinositide	NN	O	B-protein
3-kinase	NN	O	I-protein
with	NN	O	O
CD28	NN	O	B-protein
and	NN	O	O
enhanced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
phospholipase	NN	O	O
Cgamma	NN	O	O
were	NN	O	O
seen	NN	O	O
after	NN	O	O
anti-CD28	NN	O	O
mAb	NN	O	O
and	NN	O	O
CHO-CD80	NN	O	O
stimulation	NN	O	O
but	NN	O	O
to	NN	O	O
a	NN	O	O
much	NN	O	O
lesser	NN	O	O
extent	NN	O	O
after	NN	O	O
CHO-CD86	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
ligation	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
with	NN	O	O
either	NN	O	O
CD80	NN	O	B-protein
or	NN	O	O
CD86	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
shared	NN	O	O
early	NN	O	O
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CBL	NN	O	B-protein
and	NN	O	O
VAV	NN	O	B-protein
,	NN	O	O
to	NN	O	O
NFAT	NN	O	B-protein
-mediated	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
the	NN	O	O
costimulation	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
and	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
production	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
CD80	NN	O	B-protein
and	NN	O	O
CD86	NN	O	B-protein
also	NN	O	O
induce	NN	O	O
distinct	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
including	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
and	NN	O	O
phospholipase	NN	O	B-protein
Cgamma1	NN	O	I-protein
and	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
-dependent	NN	O	O
association	NN	O	O
of	NN	O	O
phosphoinositide	NN	O	B-protein
3-kinase	NN	O	I-protein
with	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

These	NN	O	O
quantitative	NN	O	O
,	NN	O	O
if	NN	O	O
not	NN	O	O
qualitative	NN	O	O
,	NN	O	O
differences	NN	O	O
between	NN	O	O
signaling	NN	O	O
initiated	NN	O	O
by	NN	O	O
these	NN	O	O
two	NN	O	O
ligands	NN	O	O
for	NN	O	O
CD28	NN	O	B-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
functional	NN	O	O
differences	NN	O	O
(	NN	O	O
e.g.	NN	O	O
Th1	NN	O	O
or	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
)	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Binding	NN	O	O
of	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
elicits	NN	O	O
structural	NN	O	O
changes	NN	O	O
in	NN	O	O
a	NN	O	O
silencer	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Proteins	NN	O	O
involved	NN	O	O
in	NN	O	O
repression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
gene	NN	O	I-DNA
may	NN	O	O
be	NN	O	O
useful	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
sickle	NN	O	O
cell	NN	O	O
anemia	NN	O	O
,	NN	O	O
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
therapy	NN	O	O
to	NN	O	O
reactivate	NN	O	O
fetal	NN	O	B-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

If	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
reciprocal	NN	O	O
elevation	NN	O	O
of	NN	O	O
gamma-globin	NN	O	B-protein
expression	NN	O	O
upon	NN	O	O
repression	NN	O	O
,	NN	O	O
this	NN	O	O
approach	NN	O	O
could	NN	O	O
be	NN	O	O
useful	NN	O	O
in	NN	O	O
additional	NN	O	O
hemoglobinopathies	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
showed	NN	O	O
that	NN	O	O
repression	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	O
gene	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
through	NN	O	O
two	NN	O	O
DNA	NN	O	B-DNA
sequences	NN	O	I-DNA
,	NN	O	O
silencers	NN	O	B-DNA
I	NN	O	I-DNA
and	NN	O	I-DNA
II	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
identified	NN	O	O
a	NN	O	O
protein	NN	O	O
termed	NN	O	O
BP1	NN	O	B-protein
which	NN	O	O
binds	NN	O	O
to	NN	O	O
both	NN	O	O
silencer	NN	O	O
sequences	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
cloned	NN	O	O
two	NN	O	O
cDNAs	NN	O	B-DNA
encoding	NN	O	O
proteins	NN	O	O
which	NN	O	O
bind	NN	O	O
to	NN	O	O
an	NN	O	O
oligonucleotide	NN	O	O
in	NN	O	O
silencer	NN	O	B-DNA
I	NN	O	I-DNA
containing	NN	O	O
a	NN	O	O
BP1	NN	O	B-protein
binding	NN	O	O
site	NN	O	O
.	NN	O	O

These	NN	O	O
cDNAs	NN	O	B-DNA
correspond	NN	O	O
to	NN	O	O
HMG-I	NN	O	B-protein
and	NN	O	O
HMG-Y	NN	O	B-protein
,	NN	O	O
isoforms	NN	O	O
regarded	NN	O	O
as	NN	O	O
architectural	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
to	NN	O	O
this	NN	O	O
oligonucleotide	NN	O	O
causes	NN	O	O
bending/flexure	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
.	NN	O	O

HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
also	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
second	NN	O	O
oligonucleotide	NN	O	O
containing	NN	O	O
a	NN	O	O
BP1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
located	NN	O	O
in	NN	O	O
a	NN	O	O
negative	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
delta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
repression	NN	O	O
of	NN	O	O
adult	NN	O	O
globin	NN	O	O
genes	NN	O	O
.	NN	O	O

Expression	NN	O	O
studies	NN	O	O
revealed	NN	O	O
that	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
ubiquitously	NN	O	O
expressed	NN	O	O
in	NN	O	O
human	NN	O	O
tissues	NN	O	O
that	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
beta-globin	NN	O	B-protein
,	NN	O	O
being	NN	O	O
present	NN	O	O
in	NN	O	O
48	NN	O	O
of	NN	O	O
50	NN	O	O
tissues	NN	O	O
and	NN	O	O
six	NN	O	O
hematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
examined	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
expression	NN	O	O
is	NN	O	O
down-regulated	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
present	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
HMG-I	NN	O	B-protein
(	NN	O	I-protein
Y	NN	O	I-protein
)	NN	O	I-protein
alters	NN	O	O
DNA	NN	O	O
conformation	NN	O	O
to	NN	O	O
allow	NN	O	O
binding	NN	O	O
of	NN	O	O
repressor	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
and	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
relative	NN	O	O
amount	NN	O	O
of	NN	O	O
HMG-I	NN	O	O
(	NN	O	O
Y	NN	O	O
)	NN	O	O
helps	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
repressive	NN	O	O
state	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Interactions	NN	O	O
between	NN	O	O
the	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
and	NN	O	O
CREB	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
increase	NN	O	O
transcription	NN	O	O
of	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Class	NN	O	B-DNA
II	NN	O	I-DNA
major	NN	O	I-DNA
histocompatibility	NN	O	I-DNA
(	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
are	NN	O	O
regulated	NN	O	O
in	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	O
and	NN	O	O
gamma	NN	O	O
interferon-inducible	NN	O	O
fashion	NN	O	O
.	NN	O	O

The	NN	O	O
master	NN	O	O
switch	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
is	NN	O	O
the	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
functions	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
is	NN	O	O
to	NN	O	O
recruit	NN	O	O
the	NN	O	O
CREB	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CBP	NN	O	B-protein
)	NN	O	O
to	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

Not	NN	O	O
only	NN	O	O
functional	NN	O	O
but	NN	O	O
also	NN	O	O
specific	NN	O	O
binding	NN	O	O
interactions	NN	O	O
between	NN	O	O
CIITA	NN	O	B-protein
and	NN	O	O
CBP	NN	O	B-protein
were	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
form	NN	O	O
of	NN	O	O
CBP	NN	O	B-protein
decreased	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
and	NN	O	O
levels	NN	O	O
of	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
determinants	NN	O	I-protein
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
cells	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
hormone	NN	O	O
could	NN	O	O
be	NN	O	O
attributed	NN	O	O
to	NN	O	O
the	NN	O	O
squelching	NN	O	O
of	NN	O	O
CBP	NN	O	B-protein
by	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
CBP	NN	O	B-protein
,	NN	O	O
a	NN	O	O
histone	NN	O	B-protein
acetyltransferase	NN	O	I-protein
,	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Role	NN	O	O
of	NN	O	O
cyclic	NN	O	B-protein
AMP	NN	O	I-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
NF-kappaB	NN	O	B-protein
family	NN	O	O
of	NN	O	O
transcription	NN	O	O
factors	NN	O	O
regulates	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
genes	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
elevation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
inhibits	NN	O	O
NF-kappaB	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
without	NN	O	O
preventing	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
examined	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
this	NN	O	O
inhibition	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
signaling	NN	O	O
pathway	NN	O	O
may	NN	O	O
inhibit	NN	O	O
NF-kappaB	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
by	NN	O	O
phosphorylating	NN	O	O
proteins	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
cAMP	NN	O	B-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
compete	NN	O	O
for	NN	O	O
limiting	NN	O	O
amounts	NN	O	O
of	NN	O	O
the	NN	O	O
coactivator	NN	O	O
CBP	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
amino-terminal	NN	O	B-protein
region	NN	O	I-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
1-450	NN	O	I-protein
)	NN	O	O
of	NN	O	O
CBP	NN	O	B-protein
specifically	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
carboxyl-terminal	NN	O	B-protein
region	NN	O	I-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
286-551	NN	O	I-protein
)	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
p65	NN	O	B-protein
(	NN	O	O
RelA	NN	O	B-protein
)	NN	O	O
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Functional	NN	O	O
studies	NN	O	O
using	NN	O	O
human	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
demonstrated	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
CBP	NN	O	B-protein
rescued	NN	O	O
cAMP	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
and	NN	O	O
transcription	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
chimeric	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
GAL4-p65	NN	O	B-protein
(	NN	O	I-protein
286-551	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
which	NN	O	O
contained	NN	O	O
the	NN	O	O
GAL4	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
fused	NN	O	O
to	NN	O	O
the	NN	O	O
carboxyl-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
p65	NN	O	B-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
286-551	NN	O	I-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
inhibited	NN	O	O
GAL4	NN	O	B-protein
-p65	NN	O	O
(	NN	O	O
286-551	NN	O	O
)	NN	O	O
-mediated	NN	O	O
transcription	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
pathway	NN	O	O
inhibits	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	O
by	NN	O	O
phosphorylating	NN	O	O
CREB	NN	O	B-protein
,	NN	O	O
which	NN	O	O
competes	NN	O	O
with	NN	O	O
p65	NN	O	B-protein
for	NN	O	O
limiting	NN	O	O
amounts	NN	O	O
of	NN	O	O
CBP	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
cord	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
healthy	NN	O	O
neonates	NN	O	O
and	NN	O	O
of	NN	O	O
preterms	NN	O	O
suffering	NN	O	O
from	NN	O	O
respiratory	NN	O	O
distress	NN	O	O
syndrome	NN	O	O
.	NN	O	O

We	NN	O	O
measured	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
cord	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
binding	NN	O	O
affinity	NN	O	O
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
in	NN	O	O
15	NN	O	O
term	NN	O	O
and	NN	O	O
in	NN	O	O
20	NN	O	O
preterm	NN	O	O
babies	NN	O	O
.	NN	O	O

Thirteen	NN	O	O
preterms	NN	O	O
of	NN	O	O
the	NN	O	O
latter	NN	O	O
group	NN	O	O
received	NN	O	O
prenatal	NN	O	O
steroid	NN	O	O
treatment	NN	O	O
.	NN	O	O

Seven	NN	O	O
preterms	NN	O	O
developed	NN	O	O
neonatal	NN	O	O
respiratory	NN	O	O
distress	NN	O	O
syndrome	NN	O	O
(	NN	O	O
NRDS	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
and	NN	O	O
the	NN	O	O
Kd	NN	O	O
were	NN	O	O
similar	NN	O	O
in	NN	O	O
the	NN	O	O
term	NN	O	O
and	NN	O	O
preterm	NN	O	O
(	NN	O	O
with	NN	O	O
and	NN	O	O
without	NN	O	O
NRDS	NN	O	O
)	NN	O	O
babies	NN	O	O
.	NN	O	O

The	NN	O	O
maximum	NN	O	O
(	NN	O	O
3H	NN	O	O
)	NN	O	O
-thymidine	NN	O	O
incorporation	NN	O	O
into	NN	O	O
DNA	NN	O	O
of	NN	O	O
cord	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
all	NN	O	O
preterms	NN	O	O
,	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
NRDS	NN	O	O
was	NN	O	O
suppressed	NN	O	O
when	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
from	NN	O	O
term	NN	O	O
babies	NN	O	O
or	NN	O	O
adults	NN	O	O
.	NN	O	O

This	NN	O	O
could	NN	O	O
partly	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
the	NN	O	O
antenatal	NN	O	O
steroid	NN	O	O
treatment	NN	O	O
.	NN	O	O

Sensitivity	NN	O	O
(	NN	O	O
ID50	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
for	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
the	NN	O	O
same	NN	O	O
in	NN	O	O
all	NN	O	O
groups	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
on	NN	O	O
the	NN	O	O
number	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
we	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
find	NN	O	O
a	NN	O	O
relation	NN	O	O
between	NN	O	O
the	NN	O	O
functionality	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
and	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
NRDS	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inefficient	NN	O	O
termination	NN	O	O
of	NN	O	O
antigen	NN	O	O
responses	NN	O	O
in	NN	O	O
NF-ATp	NN	O	B-protein
-deficient	NN	O	O
mice	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-ATp	NN	O	B-protein
,	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
prominent	NN	O	O
members	NN	O	O
of	NN	O	O
family	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
we	NN	O	O
created	NN	O	O
NF-ATp	NN	O	B-protein
-deficient	NN	O	O
mice	NN	O	O
by	NN	O	O
gene	NN	O	O
targeting	NN	O	O
.	NN	O	O

Such	NN	O	O
NF-ATp	NN	O	B-protein
-/-	NN	O	O
mice	NN	O	O
are	NN	O	O
born	NN	O	O
and	NN	O	O
appear	NN	O	O
to	NN	O	O
develop	NN	O	O
a	NN	O	O
normal	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Apart	NN	O	O
from	NN	O	O
clear-cut	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
mRNAs	NN	O	B-RNA
for	NN	O	O
Th2-type	NN	O	B-protein
lymphokines	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-5	NN	O	B-protein
,	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
in	NN	O	O
primary	NN	O	O
and	NN	O	O
secondary	NN	O	O
stimulations	NN	O	O
of	NN	O	O
spleen	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
of	NN	O	O
a	NN	O	O
distinct	NN	O	O
impaired	NN	O	O
deletion	NN	O	O
of	NN	O	O
V	NN	O	B-cell_line
beta	NN	O	I-cell_line
11+/CD4+	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
from	NN	O	O
these	NN	O	O
mice	NN	O	O
was	NN	O	O
detected	NN	O	O
after	NN	O	O
superantigen	NN	O	O
injection	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
NF-ATp	NN	O	B-protein
-/-	NN	O	O
mice	NN	O	O
older	NN	O	O
than	NN	O	O
6	NN	O	O
weeks	NN	O	O
show	NN	O	O
an	NN	O	O
2-5	NN	O	O
fold	NN	O	O
increase	NN	O	O
in	NN	O	O
number	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

This	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
activation	NN	O	B-protein
markers	NN	O	I-protein
CD44	NN	O	B-protein
and	NN	O	O
CD69	NN	O	B-protein
and	NN	O	O
decreased	NN	O	O
expression	NN	O	O
of	NN	O	O
CD62	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

AML	NN	O	B-protein
and	NN	O	I-protein
Ets	NN	O	I-protein
proteins	NN	O	I-protein
regulate	NN	O	O
the	NN	O	O
I	NN	O	B-DNA
alpha1	NN	O	I-DNA
germ-line	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
immunoglobulin	NN	O	B-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
IgH	NN	O	B-protein
)	NN	O	O
class	NN	O	O
switch	NN	O	O
recombination	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
preferentially	NN	O	O
targets	NN	O	O
unrearranged	NN	O	O
IgH	NN	O	B-DNA
genes	NN	O	I-DNA
that	NN	O	O
have	NN	O	O
already	NN	O	O
been	NN	O	O
rendered	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
.	NN	O	O

Transcription	NN	O	O
of	NN	O	O
the	NN	O	O
germ-line	NN	O	B-DNA
IgH	NN	O	I-DNA
genes	NN	O	I-DNA
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
intervening	NN	O	B-DNA
(	NN	O	I-DNA
I	NN	O	I-DNA
)	NN	O	I-DNA
regions	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
their	NN	O	O
switch	NN	O	O
regions	NN	O	O
.	NN	O	O

The	NN	O	O
I	NN	O	O
alpha1	NN	O	O
promoter	NN	O	O
activates	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
germ-line	NN	O	I-DNA
C	NN	O	I-DNA
alpha1	NN	O	I-DNA
gene	NN	O	I-DNA
for	NN	O	O
IgA1	NN	O	B-protein
and	NN	O	O
mediates	NN	O	O
the	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TGF	NN	O	I-protein
)	NN	O	I-protein
-beta1	NN	O	I-protein
responsiveness	NN	O	O
of	NN	O	O
this	NN	O	O
locus	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
I	NN	O	O
alpha1	NN	O	O
promoter	NN	O	O
contains	NN	O	O
several	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
AML/PEBP2/CBF	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
and	NN	O	O
that	NN	O	O
AML	NN	O	B-protein
and	NN	O	I-protein
Ets	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
major	NN	O	O
regulators	NN	O	O
of	NN	O	O
the	NN	O	O
basal	NN	O	O
and	NN	O	O
TGF-beta-inducible	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
constitute	NN	O	O
a	NN	O	O
starting	NN	O	O
point	NN	O	O
for	NN	O	O
studies	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
TGF-beta	NN	O	B-protein
regulates	NN	O	O
IgA	NN	O	B-protein
production	NN	O	O
.	NN	O	O

-DOCSTART-	O

Daidzein	NN	O	O
and	NN	O	O
genistein	NN	O	O
glucuronides	NN	O	O
in	NN	O	O
vitro	NN	O	O
are	NN	O	O
weakly	NN	O	O
estrogenic	NN	O	O
and	NN	O	O
activate	NN	O	O
human	NN	O	B-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
nutritionally	NN	O	O
relevant	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Daidzein	NN	O	O
and	NN	O	O
genistein	NN	O	O
glucuronides	NN	O	O
(	NN	O	O
DG	NN	O	O
and	NN	O	O
GG	NN	O	O
)	NN	O	O
,	NN	O	O
major	NN	O	O
isoflavone	NN	O	O
metabolites	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
partly	NN	O	O
responsible	NN	O	O
for	NN	O	O
biological	NN	O	O
effects	NN	O	O
of	NN	O	O
isoflavones	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
estrogen	NN	O	O
receptor	NN	O	O
binding	NN	O	O
and	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cell	NN	O	I-cell_type
(	NN	O	O
NK	NN	O	O
)	NN	O	O
activation	NN	O	O
or	NN	O	O
inhibition	NN	O	O
.	NN	O	O

DG	NN	O	O
and	NN	O	O
GG	NN	O	O
were	NN	O	O
synthesized	NN	O	O
using	NN	O	O
3-methylcholanthrene-induced	NN	O	O
rat	NN	O	O
liver	NN	O	O
microsomes	NN	O	O
.	NN	O	O

The	NN	O	O
Km	NN	O	O
and	NN	O	O
Vmax	NN	O	O
for	NN	O	O
daidzein	NN	O	O
and	NN	O	O
genistein	NN	O	O
were	NN	O	O
9.0	NN	O	O
and	NN	O	O
7.7	NN	O	O
micromol/L	NN	O	O
,	NN	O	O
and	NN	O	O
0.7	NN	O	O
and	NN	O	O
1.6	NN	O	O
micromol/	NN	O	O
(	NN	O	O
mg	NN	O	O
protein.	NN	O	O
min	NN	O	O
)	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
absence	NN	O	O
of	NN	O	O
ultraviolet	NN	O	O
absorbance	NN	O	O
maxima	NN	O	O
shifts	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
sodium	NN	O	O
acetate	NN	O	O
confirmed	NN	O	O
that	NN	O	O
the	NN	O	O
synthesized	NN	O	O
products	NN	O	O
were	NN	O	O
7-O-glucuronides	NN	O	O
.	NN	O	O

DG	NN	O	O
and	NN	O	O
GG	NN	O	O
were	NN	O	O
further	NN	O	O
purified	NN	O	O
by	NN	O	O
a	NN	O	O
Sephadex	NN	O	O
LH-20	NN	O	O
column	NN	O	O
.	NN	O	O

DG	NN	O	O
and	NN	O	O
GG	NN	O	O
competed	NN	O	O
with	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
17beta-	NN	O	O
(	NN	O	O
3H	NN	O	O
)	NN	O	O
estradiol	NN	O	O
to	NN	O	O
estrogen	NN	O	B-protein
receptors	NN	O	I-protein
of	NN	O	O
B6D2F1	NN	O	O
mouse	NN	O	O
uterine	NN	O	O
cytosol	NN	O	O
.	NN	O	O

The	NN	O	O
concentrations	NN	O	O
required	NN	O	O
for	NN	O	O
50	NN	O	O
%	NN	O	O
displacement	NN	O	O
of	NN	O	O
17beta-	NN	O	O
(	NN	O	O
3H	NN	O	O
)	NN	O	O
estradiol	NN	O	O
(	NN	O	O
CB50	NN	O	O
)	NN	O	O
were	NN	O	O
:	NN	O	O
17beta-estradiol	NN	O	O
,	NN	O	O
1.34	NN	O	O
nmol/L	NN	O	O
;	NN	O	O
diethylstilbestrol	NN	O	O
,	NN	O	O
1.46	NN	O	O
nmol/L	NN	O	O
;	NN	O	O
daidzein	NN	O	O
,	NN	O	O
1.6	NN	O	O
micromol/L	NN	O	O
;	NN	O	O
DG	NN	O	O
,	NN	O	O
14.7	NN	O	O
micromol/L	NN	O	O
;	NN	O	O
genistein	NN	O	O
,	NN	O	O
0.154	NN	O	O
micromol/L	NN	O	O
;	NN	O	O
GG	NN	O	O
,	NN	O	O
7.27	NN	O	O
micromol/L	NN	O	O
.	NN	O	O

In	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
genistein	NN	O	O
at	NN	O	O
<	NN	O	O
0.5	NN	O	O
micromol/L	NN	O	O
and	NN	O	O
DG	NN	O	O
and	NN	O	O
GG	NN	O	O
at	NN	O	O
0.1-10	NN	O	O
micromol/L	NN	O	O
enhanced	NN	O	O
NK	NN	O	O
cell-mediated	NN	O	O
K562	NN	O	B-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
killing	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

At	NN	O	O
>	NN	O	O
0.5	NN	O	O
micromol/L	NN	O	O
,	NN	O	O
genistein	NN	O	O
inhibited	NN	O	O
NK	NN	O	O
cytotoxicity	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
glucuronides	NN	O	O
only	NN	O	O
inhibited	NN	O	O
NK	NN	O	O
cytotoxicity	NN	O	O
at	NN	O	O
50	NN	O	O
micromol/L	NN	O	O
.	NN	O	O

Isoflavones	NN	O	O
,	NN	O	O
and	NN	O	O
especially	NN	O	O
the	NN	O	O
isoflavone	NN	O	O
glucuronides	NN	O	O
,	NN	O	O
enhanced	NN	O	O
activation	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
additively	NN	O	O
.	NN	O	O

At	NN	O	O
physiological	NN	O	O
concentrations	NN	O	O
,	NN	O	O
DG	NN	O	O
and	NN	O	O
GG	NN	O	O
were	NN	O	O
weakly	NN	O	O
estrogenic	NN	O	O
,	NN	O	O
and	NN	O	O
they	NN	O	O
activated	NN	O	O
human	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
nutritionally	NN	O	O
relevant	NN	O	O
concentrations	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
probably	NN	O	O
at	NN	O	O
a	NN	O	O
site	NN	O	O
different	NN	O	O
from	NN	O	O
IL-2	NN	O	B-protein
action	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cross-linking	NN	O	O
of	NN	O	O
CD44	NN	O	B-protein
on	NN	O	O
rheumatoid	NN	O	B-cell_type
synovial	NN	O	I-cell_type
cells	NN	O	I-cell_type
up-regulates	NN	O	O
VCAM-1	NN	O	B-protein
.	NN	O	O

CD44	NN	O	O
is	NN	O	O
a	NN	O	O
ubiquitous	NN	O	O
molecule	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
hyaluronic	NN	O	O
acid	NN	O	O
or	NN	O	O
homing	NN	O	O
receptor	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
cellular	NN	O	O
functions	NN	O	O
and	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
inflammation	NN	O	O
,	NN	O	O
for	NN	O	O
example	NN	O	O
,	NN	O	O
rheumatoid	NN	O	O
synovitis	NN	O	O
,	NN	O	O
are	NN	O	O
currently	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
a	NN	O	O
novel	NN	O	O
function	NN	O	O
for	NN	O	O
CD44	NN	O	O
.	NN	O	O

Using	NN	O	O
synovial	NN	O	O
cells	NN	O	O
from	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
patients	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
CD44	NN	O	O
cross-linking	NN	O	O
and	NN	O	O
binding	NN	O	O
to	NN	O	O
hyaluronan	NN	O	O
augmented	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
subsequently	NN	O	O
VCAM-1	NN	O	B-protein
-mediated	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
.	NN	O	O

Briefly	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
1	NN	O	O
)	NN	O	O
rheumatoid	NN	O	B-cell_type
synovial	NN	O	I-cell_type
cells	NN	O	I-cell_type
highly	NN	O	O
expressed	NN	O	O
CD44	NN	O	B-protein
;	NN	O	O
2	NN	O	O
)	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
CD44	NN	O	B-protein
markedly	NN	O	O
but	NN	O	O
transiently	NN	O	O
augmented	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
its	NN	O	O
mRNA	NN	O	O
transcription	NN	O	O
much	NN	O	O
more	NN	O	O
than	NN	O	O
did	NN	O	O
IL-1beta	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	O
;	NN	O	O
3	NN	O	O
)	NN	O	O
hyaluronan	NN	O	O
,	NN	O	O
especially	NN	O	O
when	NN	O	O
fragmented	NN	O	O
,	NN	O	O
also	NN	O	O
up-regulated	NN	O	O
VCAM-1	NN	O	B-protein
;	NN	O	O
4	NN	O	O
)	NN	O	O
CD44	NN	O	B-protein
activated	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
AP-1	NN	O	O
;	NN	O	O
and	NN	O	O
5	NN	O	O
)	NN	O	O
the	NN	O	O
integrin	NN	O	B-protein
-dependent	NN	O	O
adhesive	NN	O	O
function	NN	O	O
of	NN	O	O
RA	NN	O	B-cell_type
synovial	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
also	NN	O	O
amplified	NN	O	O
by	NN	O	O
CD44	NN	O	B-protein
cross-linking	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
adhesion	NN	O	O
of	NN	O	O
RA	NN	O	B-cell_type
synovial	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
matrices	NN	O	O
such	NN	O	O
as	NN	O	O
hyaluronic	NN	O	O
acid	NN	O	O
through	NN	O	O
CD44	NN	O	B-protein
could	NN	O	O
up-regulate	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
VCAM-1	NN	O	B-protein
-mediated	NN	O	O
adhesion	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
might	NN	O	O
in	NN	O	O
turn	NN	O	O
cause	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
synovial	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
RA	NN	O	O
synovitis	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
propose	NN	O	O
that	NN	O	O
such	NN	O	O
cross-talking	NN	O	O
among	NN	O	O
distinct	NN	O	O
adhesion	NN	O	O
molecules	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
inflammation	NN	O	O
,	NN	O	O
including	NN	O	O
RA	NN	O	O
synovitis	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappaB	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
regulator	NN	O	O
of	NN	O	O
human	NN	O	O
granulocyte	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

During	NN	O	O
beneficial	NN	O	O
inflammation	NN	O	O
,	NN	O	O
potentially	NN	O	O
tissue-damaging	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
undergo	NN	O	O
apoptosis	NN	O	O
before	NN	O	O
being	NN	O	O
cleared	NN	O	O
by	NN	O	O
phagocytes	NN	O	O
in	NN	O	O
a	NN	O	O
non-phlogistic	NN	O	O
manner	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
constitutive	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
and	NN	O	O
eosinophils	NN	O	B-cell_type
is	NN	O	O
greatly	NN	O	O
accelerated	NN	O	O
in	NN	O	O
both	NN	O	O
a	NN	O	O
rapid	NN	O	O
and	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
by	NN	O	O
the	NN	O	O
fungal	NN	O	O
metabolite	NN	O	O
gliotoxin	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
its	NN	O	O
inactive	NN	O	O
analog	NN	O	O
methylthiogliotoxin	NN	O	O
.	NN	O	O

This	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
abolished	NN	O	O
by	NN	O	O
the	NN	O	O
caspase	NN	O	O
inhibitor	NN	O	O
zVAD-fmk	NN	O	O
,	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
mimicked	NN	O	O
by	NN	O	O
a	NN	O	O
cell	NN	O	O
permeable	NN	O	O
inhibitory	NN	O	O
peptide	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
SN-50	NN	O	O
;	NN	O	O
other	NN	O	O
NF-kappaB	NN	O	B-protein
inhibitors	NN	O	O
,	NN	O	O
curcumin	NN	O	O
and	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
;	NN	O	O
and	NN	O	O
the	NN	O	O
proteasome	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
MG-132	NN	O	O
.	NN	O	O

Gliotoxin	NN	O	O
also	NN	O	O
augmented	NN	O	O
dramatically	NN	O	O
the	NN	O	O
early	NN	O	O
(	NN	O	O
2-6	NN	O	O
h	NN	O	O
)	NN	O	O
pro-apoptotic	NN	O	O
effects	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
unmasked	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
eosinophil	NN	O	B-cell_type
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
TNF-alpha	NN	O	B-protein
caused	NN	O	O
a	NN	O	O
gliotoxin-inhibitable	NN	O	O
activation	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	O
form	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
a	NN	O	O
response	NN	O	O
that	NN	O	O
may	NN	O	O
underlie	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
to	NN	O	O
delay	NN	O	O
apoptosis	NN	O	O
at	NN	O	O
later	NN	O	O
times	NN	O	O
(	NN	O	O
12-24	NN	O	O
h	NN	O	O
)	NN	O	O
and	NN	O	O
limit	NN	O	O
its	NN	O	O
early	NN	O	O
killing	NN	O	O
effect	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
cycloheximide	NN	O	O
displayed	NN	O	O
a	NN	O	O
similar	NN	O	O
capacity	NN	O	O
to	NN	O	O
enhance	NN	O	O
TNF-alpha	NN	O	B-protein
induced	NN	O	O
neutrophil	NN	O	O
apoptosis	NN	O	O
even	NN	O	O
at	NN	O	O
time	NN	O	O
points	NN	O	O
when	NN	O	O
cycloheximide	NN	O	O
alone	NN	O	O
had	NN	O	O
no	NN	O	O
pro-apoptotic	NN	O	O
effect	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
may	NN	O	O
regulate	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
protein	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
which	NN	O	O
protect	NN	O	O
neutrophils	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
cytotoxic	NN	O	O
effects	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
shed	NN	O	O
light	NN	O	O
on	NN	O	O
the	NN	O	O
biochemical	NN	O	O
and	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
regulating	NN	O	O
human	NN	O	O
granulocyte	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
physiological	NN	O	O
cell	NN	O	O
death	NN	O	O
pathway	NN	O	O
in	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Role	NN	O	O
of	NN	O	O
Egr-2	NN	O	B-protein
in	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
in	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
aberrant	NN	O	B-cell_type
double-negative	NN	O	I-cell_type
lpr	NN	O	I-cell_type
and	NN	O	I-cell_type
gld	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
previously	NN	O	O
identified	NN	O	O
a	NN	O	O
Fas	NN	O	B-DNA
ligand	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
FLRE	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
Fas	NN	O	B-DNA
ligand	NN	O	I-DNA
(	NN	O	I-DNA
fasL	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
that	NN	O	O
binds	NN	O	O
Egr	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
Egr-3	NN	O	B-protein
(	NN	O	O
PILOT	NN	O	B-protein
)	NN	O	O
but	NN	O	O
not	NN	O	O
Egr-1	NN	O	B-protein
(	NN	O	O
NGFI-A	NN	O	B-protein
,	NN	O	O
Krox-24	NN	O	B-protein
,	NN	O	O
Tis-8	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Zif-268	NN	O	B-protein
)	NN	O	O
induces	NN	O	O
transcription	NN	O	O
of	NN	O	O
fasL	NN	O	B-protein
.	NN	O	O

The	NN	O	O
aberrant	NN	O	B-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
-	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
lpr/lpr	NN	O	O
and	NN	O	O
gld/gld	NN	O	O
mice	NN	O	O
,	NN	O	O
which	NN	O	O
have	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
genes	NN	O	O
encoding	NN	O	O
Fas	NN	O	B-protein
and	NN	O	O
FasL	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
have	NN	O	O
an	NN	O	O
activated	NN	O	O
phenotype	NN	O	O
and	NN	O	O
constitutively	NN	O	O
express	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
fasL	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
prompting	NN	O	O
us	NN	O	O
to	NN	O	O
ask	NN	O	O
what	NN	O	O
role	NN	O	O
if	NN	O	O
any	NN	O	O
the	NN	O	O
FLRE	NN	O	B-DNA
and	NN	O	O
Egr	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
have	NN	O	O
in	NN	O	O
this	NN	O	O
aberrant	NN	O	O
expression	NN	O	O
of	NN	O	O
fasL	NN	O	B-protein
.	NN	O	O

Unstimulated	NN	O	O
MRL-lpr/lpr	NN	O	B-cell_line
and	NN	O	I-cell_line
C3H-gld/gld	NN	O	I-cell_line
CD4	NN	O	I-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
CD8	NN	O	I-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
constitutively	NN	O	O
contained	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
two	NN	O	O
proteins	NN	O	O
that	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
FLRE	NN	O	B-DNA
.	NN	O	O

Supershift	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
these	NN	O	O
proteins	NN	O	O
to	NN	O	O
be	NN	O	O
Egr-1	NN	O	B-protein
and	NN	O	O
Egr-2	NN	O	B-protein
(	NN	O	O
Krox-20	NN	O	O
)	NN	O	O
;	NN	O	O
Egr-3	NN	O	B-protein
was	NN	O	O
not	NN	O	O
detected	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
lymph	NN	O	I-cell_type
node	NN	O	I-cell_type
cells	NN	O	I-cell_type
resulted	NN	O	O
in	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
,	NN	O	I-protein
-2	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
-3	NN	O	I-protein
.	NN	O	O

As	NN	O	O
with	NN	O	O
egr-3	NN	O	B-DNA
,	NN	O	O
expression	NN	O	O
of	NN	O	O
egr-2	NN	O	B-DNA
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

Although	NN	O	O
overexpressed	NN	O	O
Egr-1	NN	O	B-protein
was	NN	O	O
ineffective	NN	O	O
,	NN	O	O
overexpressed	NN	O	O
Egr-2	NN	O	B-protein
was	NN	O	O
as	NN	O	O
potent	NN	O	O
as	NN	O	O
Egr-3	NN	O	B-protein
in	NN	O	O
inducing	NN	O	O
fasL	NN	O	B-DNA
promoter-dependent	NN	O	I-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
T	NN	O	O
cell	NN	O	O
hybridomas	NN	O	O
and	NN	O	O
HeLa	NN	O	O
cells	NN	O	O
,	NN	O	O
and	NN	O	O
both	NN	O	O
up-regulated	NN	O	O
endogenous	NN	O	O
fasL	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

FasL-dependent	NN	O	B-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
MRL-lpr/lpr	NN	O	B-cell_line
and	NN	O	I-cell_line
C3H-gld/gld	NN	O	I-cell_line
CD4	NN	O	I-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
CD8	NN	O	I-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
constitutively	NN	O	O
active	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
activity	NN	O	O
was	NN	O	O
largely	NN	O	O
prevented	NN	O	O
by	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
critical	NN	O	O
Egr	NN	O	B-DNA
family	NN	O	I-DNA
binding	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Egr-2	NN	O	B-protein
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
Egr-3	NN	O	B-protein
,	NN	O	O
regulates	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
activated	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
Egr-2	NN	O	B-protein
is	NN	O	O
likely	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
direct	NN	O	O
role	NN	O	O
in	NN	O	O
aberrant	NN	O	O
fasL	NN	O	B-protein
up-regulation	NN	O	O
in	NN	O	O
lpr/lpr	NN	O	B-cell_line
and	NN	O	I-cell_line
gld/gld	NN	O	I-cell_line
CD4	NN	O	I-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
CD8	NN	O	I-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
CCAAT-box	NN	O	B-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
NF-Y	NN	O	I-protein
is	NN	O	O
modulated	NN	O	O
through	NN	O	O
the	NN	O	O
regulated	NN	O	O
expression	NN	O	O
of	NN	O	O
its	NN	O	O
A	NN	O	B-protein
subunit	NN	O	I-protein
during	NN	O	O
monocyte	NN	O	O
to	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
:	NN	O	O
regulation	NN	O	O
of	NN	O	O
tissue-specific	NN	O	B-DNA
genes	NN	O	I-DNA
through	NN	O	O
a	NN	O	O
ubiquitous	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF-Y	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
human	NN	O	O
monocyte	NN	O	O
to	NN	O	O
macrophage	NN	O	O
maturation	NN	O	O
.	NN	O	O

NF-Y	NN	O	B-protein
is	NN	O	O
a	NN	O	O
ubiquitous	NN	O	O
and	NN	O	O
evolutionarily	NN	O	O
conserved	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
specifically	NN	O	O
to	NN	O	O
the	NN	O	O
CCAAT	NN	O	B-DNA
motif	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
genes	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
in	NN	O	O
circulating	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
NF-Y	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
not	NN	O	O
detected	NN	O	O
on	NN	O	O
the	NN	O	O
CCAAT	NN	O	B-DNA
motif	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	I-DNA
genes	NN	O	I-DNA
such	NN	O	O
as	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
(	NN	O	I-DNA
MHC	NN	O	I-DNA
)	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
,	NN	O	O
gp91-phox	NN	O	B-DNA
,	NN	O	O
mig	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
fibronectin	NN	O	O
,	NN	O	O
whereas	NN	O	O
during	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
,	NN	O	O
a	NN	O	O
progressive	NN	O	O
increase	NN	O	O
in	NN	O	O
NF-Y	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
observed	NN	O	O
on	NN	O	O
these	NN	O	O
promoters	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
NF-Y	NN	O	B-protein
subunit	NN	O	O
expression	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
NF-Y	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
circulating	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
is	NN	O	O
caused	NN	O	O
by	NN	O	O
a	NN	O	O
lack	NN	O	O
of	NN	O	O
the	NN	O	O
A	NN	O	B-protein
subunit	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
the	NN	O	O
recombinant	NN	O	O
NF-Y	NN	O	B-protein
A	NN	O	B-protein
subunit	NN	O	I-protein
restores	NN	O	O
NF-Y	NN	O	B-protein
binding	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
NF-YA	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
due	NN	O	O
to	NN	O	O
posttranscriptional	NN	O	O
regulation	NN	O	O
and	NN	O	O
not	NN	O	O
to	NN	O	O
a	NN	O	O
specific	NN	O	O
proteolytic	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
NF-YA	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
present	NN	O	O
at	NN	O	O
the	NN	O	O
same	NN	O	O
level	NN	O	O
at	NN	O	O
all	NN	O	O
days	NN	O	O
of	NN	O	O
monocyte	NN	O	O
cultivation	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
protein	NN	O	O
is	NN	O	O
absent	NN	O	O
in	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
but	NN	O	O
is	NN	O	O
progressively	NN	O	O
synthesized	NN	O	O
during	NN	O	O
the	NN	O	O
maturation	NN	O	O
process	NN	O	O
.	NN	O	O

We	NN	O	O
thus	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
NF-Y	NN	O	B-protein
A	NN	O	B-protein
subunit	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
relevant	NN	O	O
role	NN	O	O
in	NN	O	O
activating	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
highly	NN	O	O
expressed	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
line	NN	O	O
with	NN	O	O
this	NN	O	O
conclusion	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
cut/CDP	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
repressor	NN	O	O
that	NN	O	O
inhibits	NN	O	O
gpc91-phox	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
preventing	NN	O	O
NF-Y	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
CAAT	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
absent	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Interdomain	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
ZAP-70	NN	O	B-protein
regulates	NN	O	O
but	NN	O	O
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
ZAP-70	NN	O	B-protein
signaling	NN	O	O
function	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
ZAP-70	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
and	NN	O	O
development	NN	O	O
.	NN	O	O

After	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
stimulation	NN	O	O
,	NN	O	O
ZAP-70	NN	O	B-protein
associates	NN	O	O
with	NN	O	O
the	NN	O	O
receptor	NN	O	O
and	NN	O	O
is	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
many	NN	O	O
tyrosines	NN	O	O
,	NN	O	O
including	NN	O	O
Y292	NN	O	O
,	NN	O	O
Y315	NN	O	O
,	NN	O	O
and	NN	O	O
Y319	NN	O	O
within	NN	O	O
interdomain	NN	O	O
B	NN	O	O
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
Y292	NN	O	O
negatively	NN	O	O
regulates	NN	O	O
ZAP-70	NN	O	B-protein
function	NN	O	O
and	NN	O	O
that	NN	O	O
Y315	NN	O	O
positively	NN	O	O
regulates	NN	O	O
ZAP-70	NN	O	B-protein
function	NN	O	O
by	NN	O	O
interacting	NN	O	O
with	NN	O	O
Vav	NN	O	B-protein
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
Y319	NN	O	O
also	NN	O	O
positively	NN	O	O
regulate	NN	O	O
ZAP-70	NN	O	B-protein
function	NN	O	O
.	NN	O	O

Paradoxically	NN	O	O
,	NN	O	O
removal	NN	O	O
of	NN	O	O
interdomain	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
to	NN	O	O
create	NN	O	O
the	NN	O	O
construct	NN	O	O
designated	NN	O	O
Delta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
containing	NN	O	O
the	NN	O	O
Y292	NN	O	O
,	NN	O	O
Y315	NN	O	O
,	NN	O	O
and	NN	O	O
Y319	NN	O	O
sites	NN	O	O
,	NN	O	O
did	NN	O	O
not	NN	O	O
eliminate	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
ZAP-70	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
multiple	NN	O	B-DNA
gene	NN	O	I-DNA
reporters	NN	O	I-DNA
in	NN	O	O
Syk-deficient	NN	O	B-cell_line
DT-40	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
ZAP-70/Syk-deficient	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
Delta	NN	O	B-protein
still	NN	O	O
utilizes	NN	O	O
the	NN	O	O
same	NN	O	O
pathways	NN	O	O
as	NN	O	O
wild-type	NN	O	O
ZAP-70	NN	O	B-protein
to	NN	O	O
mediate	NN	O	O
NF-AT	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
manifested	NN	O	O
by	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
Delta	NN	O	B-protein
to	NN	O	O
restore	NN	O	O
induction	NN	O	O
of	NN	O	O
calcium	NN	O	O
fluxes	NN	O	O
and	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
dominant	NN	O	O
negative	NN	O	O
Ras	NN	O	B-protein
and	NN	O	O
FK506	NN	O	B-protein
to	NN	O	O
block	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
activity	NN	O	O
mediated	NN	O	O
by	NN	O	O
Delta	NN	O	B-protein
.	NN	O	O

Biochemically	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
stimulated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Vav	NN	O	B-protein
,	NN	O	O
Shc	NN	O	B-protein
,	NN	O	O
and	NN	O	O
ZAP-70	NN	O	B-protein
itself	NN	O	O
is	NN	O	O
diminished	NN	O	O
,	NN	O	O
whereas	NN	O	O
that	NN	O	O
of	NN	O	O
Slp-76	NN	O	B-protein
is	NN	O	O
increased	NN	O	O
in	NN	O	O
cells	NN	O	O
reconstituted	NN	O	O
with	NN	O	O
Delta	NN	O	B-protein
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
interdomain	NN	O	B-protein
B	NN	O	I-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
ZAP-70	NN	O	B-protein
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
receptor	NN	O	O
.	NN	O	O

The	NN	O	O
in	NN	O	O
vitro	NN	O	O
kinase	NN	O	O
activity	NN	O	O
of	NN	O	O
ZAP-70	NN	O	B-protein
lacking	NN	O	O
interdomain	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
kinase	NN	O	O
activity	NN	O	O
was	NN	O	O
still	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
protein	NN	O	O
's	NN	O	O
in	NN	O	O
vivo	NN	O	O
activity	NN	O	O
.	NN	O	O

Based	NN	O	O
on	NN	O	O
these	NN	O	O
data	NN	O	O
,	NN	O	O
we	NN	O	O
concluded	NN	O	O
that	NN	O	O
interdomain	NN	O	B-protein
B	NN	O	I-protein
regulates	NN	O	O
but	NN	O	O
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
ZAP-70	NN	O	B-protein
signaling	NN	O	O
function	NN	O	O
leading	NN	O	O
to	NN	O	O
cellular	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
induction	NN	O	O
of	NN	O	O
DNA-binding	NN	O	B-protein
activities	NN	O	I-protein
following	NN	O	O
CD19	NN	O	B-protein
cross-linking	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
molecule	NN	O	I-protein
CD19	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
all	NN	O	O
stages	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
,	NN	O	O
including	NN	O	O
normal	NN	O	O
plasma	NN	O	O
cells	NN	O	O
,	NN	O	O
and	NN	O	O
mediates	NN	O	O
signal	NN	O	O
transduction	NN	O	O
via	NN	O	O
interaction	NN	O	O
with	NN	O	O
cytoplasmic	NN	O	B-protein
effector	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Cross-linking	NN	O	O
CD19	NN	O	B-protein
on	NN	O	O
early	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
induces	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
CD19/Vav/phosphatidylinositol-3	NN	O	B-protein
kinase	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CD19	NN	O	B-protein
and	NN	O	O
Vav	NN	O	B-protein
,	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
Ras	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
explore	NN	O	O
the	NN	O	O
ramifications	NN	O	O
of	NN	O	O
CD19	NN	O	B-protein
signaling	NN	O	O
,	NN	O	O
the	NN	O	O
current	NN	O	O
study	NN	O	O
examined	NN	O	O
whether	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Elk-1	NN	O	B-protein
,	NN	O	O
activation	NN	O	O
of	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
or	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	O
factor-kappaB	NN	O	O
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
occurred	NN	O	O
following	NN	O	O
CD19	NN	O	B-protein
cross-linking	NN	O	O
.	NN	O	O

The	NN	O	O
cells	NN	O	O
used	NN	O	O
were	NN	O	O
the	NN	O	O
BLIN-1	NN	O	B-cell_line
pre-B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
expressing	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
mu	NN	O	I-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
associated	NN	O	O
with	NN	O	O
surrogate	NN	O	B-protein
light	NN	O	I-protein
chain	NN	O	I-protein
and	NN	O	O
the	NN	O	O
1E8	NN	O	O
immature	NN	O	O
B	NN	O	O
cell	NN	O	O
line	NN	O	O
expressing	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
mu/kappa	NN	O	I-protein
.	NN	O	O

Lysates	NN	O	O
from	NN	O	O
CD19	NN	O	B-protein
cross-linked	NN	O	O
1E8	NN	O	B-cell_line
cells	NN	O	I-cell_line
induced	NN	O	O
robust	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
an	NN	O	O
Elk-1	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
whereas	NN	O	O
no	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Elk-1	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
occurred	NN	O	O
using	NN	O	O
lysates	NN	O	O
from	NN	O	O
CD19	NN	O	B-cell_line
cross-linked	NN	O	I-cell_line
BLIN-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

An	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
employing	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
consensus	NN	O	O
oligonucleotides	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
AP-1	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
increased	NN	O	O
,	NN	O	O
while	NN	O	O
constitutive	NN	O	O
NF-kappaB	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
was	NN	O	O
not	NN	O	O
enhanced	NN	O	O
,	NN	O	O
following	NN	O	O
2	NN	O	O
h	NN	O	O
of	NN	O	O
CD19	NN	O	B-protein
cross-linking	NN	O	O
in	NN	O	O
1E8	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O
Supershift	NN	O	O
experiments	NN	O	O
revealed	NN	O	O
that	NN	O	O
JunD	NN	O	B-protein
and	NN	O	O
c-Fos	NN	O	B-protein
proteins	NN	O	I-protein
mediated	NN	O	O
anti-	NN	O	O
CD19	NN	O	B-protein
induced	NN	O	O
AP-1	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
1E8	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
CD19	NN	O	B-protein
cross-linking	NN	O	O
in	NN	O	O
BLIN-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
but	NN	O	O
had	NN	O	O
no	NN	O	O
apparent	NN	O	O
effect	NN	O	O
on	NN	O	O
AP-1	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
CD19	NN	O	B-protein
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
activates	NN	O	O
different	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
at	NN	O	O
juxtaposed	NN	O	O
stages	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
that	NN	O	O
may	NN	O	O
culminate	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
or	NN	O	O
suppression	NN	O	O
of	NN	O	O
distinct	NN	O	O
sets	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Oxidants	NN	O	O
,	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
intestinal	NN	O	O
inflammation	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
now	NN	O	O
well	NN	O	O
appreciated	NN	O	O
that	NN	O	O
chronic	NN	O	O
gut	NN	O	O
inflammation	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
enhanced	NN	O	O
production	NN	O	O
of	NN	O	O
reactive	NN	O	O
metabolites	NN	O	O
of	NN	O	O
oxygen	NN	O	O
and	NN	O	O
nitrogen	NN	O	O
.	NN	O	O

Some	NN	O	O
of	NN	O	O
these	NN	O	O
oxidants	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
modulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
genes	NN	O	O
that	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

For	NN	O	O
example	NN	O	O
,	NN	O	O
certain	NN	O	O
oxidants	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
regulates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
different	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
,	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
and	NN	O	O
enzymes	NN	O	B-protein
.	NN	O	O

Oxidants	NN	O	O
are	NN	O	O
also	NN	O	O
known	NN	O	O
to	NN	O	O
activate	NN	O	O
another	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
.	NN	O	O

This	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
composed	NN	O	O
of	NN	O	O
products	NN	O	O
from	NN	O	O
the	NN	O	O
fos	NN	O	B-protein
and	NN	O	I-protein
jun	NN	O	I-protein
proto-oncogene	NN	O	I-protein
family	NN	O	I-protein
and	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
regulating	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
oxidants	NN	O	O
are	NN	O	O
believed	NN	O	O
to	NN	O	O
promote	NN	O	O
intestinal	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cell	NN	O	I-cell_type
apoptosis	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
B-cell	NN	O	B-protein
lymphoma/leukemia-2	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
is	NN	O	O
believed	NN	O	O
to	NN	O	O
inhibit	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
in	NN	O	O
an	NN	O	O
antioxidant-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
nontoxic	NN	O	O
concentrations	NN	O	O
of	NN	O	O
reactive	NN	O	O
metabolites	NN	O	O
of	NN	O	O
oxygen	NN	O	O
and	NN	O	O
nitrogen	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
and	NN	O	O
in	NN	O	O
modulating	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
lymphoid-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Oct2	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Oct2	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
predominantly	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
plays	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
during	NN	O	O
the	NN	O	O
terminal	NN	O	O
phase	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
regulatory	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
several	NN	O	O
genes	NN	O	O
specifically	NN	O	O
expressed	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
contain	NN	O	O
functional	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
Oct2	NN	O	B-protein
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
originally	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
Oct2	NN	O	B-protein
were	NN	O	O
affected	NN	O	O
in	NN	O	O
their	NN	O	O
expression	NN	O	O
in	NN	O	O
Oct2-deficient	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
an	NN	O	O
attempt	NN	O	O
to	NN	O	O
find	NN	O	O
such	NN	O	O
elusive	NN	O	O
Oct2	NN	O	B-protein
target	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
molecular	NN	O	O
function	NN	O	O
of	NN	O	O
Oct2	NN	O	B-protein
in	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
cDNAs	NN	O	B-DNA
for	NN	O	O
Oct2	NN	O	B-protein
target	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

So	NN	O	O
far	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
five	NN	O	O
potential	NN	O	O
targets	NN	O	O
for	NN	O	O
Oct2	NN	O	B-protein
:	NN	O	O
the	NN	O	O
membrane	NN	O	B-protein
glycoprotein	NN	O	I-protein
CD36	NN	O	B-protein
,	NN	O	O
the	NN	O	O
cysteine-rich	NN	O	B-protein
secreted	NN	O	I-protein
protein	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
CRISP-3	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
mouse	NN	O	O
homolog	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
monocyte/neutrophil	NN	O	I-protein
elastase	NN	O	I-protein
inhibitor	NN	O	I-protein
(	NN	O	O
mEI	NN	O	B-protein
)	NN	O	O
and	NN	O	O
two	NN	O	O
unknown	NN	O	O
cDNA	NN	O	B-DNA
sequences	NN	O	I-DNA
Nov1	NN	O	B-DNA
and	NN	O	O
Nov2	NN	O	B-DNA
.	NN	O	O

These	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
show	NN	O	O
quite	NN	O	O
distinct	NN	O	O
expression	NN	O	O
patterns	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
addition	NN	O	O
to	NN	O	O
Oct2	NN	O	B-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
their	NN	O	O
regulation	NN	O	O
.	NN	O	O

Whereas	NN	O	O
CD36	NN	O	B-protein
and	NN	O	O
mEI	NN	O	B-protein
were	NN	O	O
expressed	NN	O	O
in	NN	O	O
all	NN	O	O
hematopoetic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
containing	NN	O	O
Oct2	NN	O	B-protein
,	NN	O	O
.	NN	O	O

CRISP-3	NN	O	B-protein
is	NN	O	O
pre-B	NN	O	O
cell-specific	NN	O	O
,	NN	O	O
Nov1	NN	O	B-DNA
is	NN	O	O
plasma	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
and	NN	O	O
Nov2	NN	O	B-DNA
is	NN	O	O
B	NN	O	O
cell-specifically	NN	O	O
expressed	NN	O	O
.	NN	O	O

-DOCSTART-	O

RFX-B	NN	O	B-DNA
is	NN	O	O
the	NN	O	O
gene	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
cause	NN	O	O
of	NN	O	O
the	NN	O	O
bare	NN	O	O
lymphocyte	NN	O	O
syndrome	NN	O	O
,	NN	O	O
an	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
immunodeficiency	NN	O	O
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
Immunity	NN	O	O
1999	NN	O	O
Mar	NN	O	O
;	NN	O	O
10	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
:	NN	O	O
399	NN	O	O
]	NN	O	O

The	NN	O	O
bare	NN	O	O
lymphocyte	NN	O	O
syndrome	NN	O	O
(	NN	O	O
BLS	NN	O	O
)	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
transcription	NN	O	O
and	NN	O	O
humoral-	NN	O	O
and	NN	O	O
cellular-mediated	NN	O	O
immune	NN	O	O
responses	NN	O	O
to	NN	O	O
foreign	NN	O	O
antigens	NN	O	O
.	NN	O	O

Three	NN	O	O
of	NN	O	O
the	NN	O	O
four	NN	O	O
BLS	NN	O	O
genetic	NN	O	O
complementation	NN	O	O
groups	NN	O	O
have	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
RFX	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
purified	NN	O	O
the	NN	O	O
RFX	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
sequenced	NN	O	O
its	NN	O	O
three	NN	O	O
subunits	NN	O	O
.	NN	O	O

The	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
smallest	NN	O	O
subunit	NN	O	O
describes	NN	O	O
a	NN	O	O
novel	NN	O	O
gene	NN	O	O
,	NN	O	O
termed	NN	O	O
RFX-B	NN	O	B-DNA
.	NN	O	O

RFX-B	NN	O	B-DNA
complements	NN	O	O
the	NN	O	O
predominant	NN	O	O
BLS	NN	O	B-protein
complementation	NN	O	I-protein
group	NN	O	I-protein
(	NN	O	O
group	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
and	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
mutant	NN	O	O
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
from	NN	O	O
this	NN	O	O
BLS	NN	O	O
group	NN	O	O
.	NN	O	O

The	NN	O	O
protein	NN	O	O
has	NN	O	O
no	NN	O	O
known	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
but	NN	O	O
does	NN	O	O
contain	NN	O	O
three	NN	O	O
ankyrin	NN	O	O
repeats	NN	O	O
that	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Binding	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
to	NN	O	O
STAT5	NN	O	B-DNA
target	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
HTLV-I-transformed	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
type	NN	O	O
I	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
induces	NN	O	O
abnormal	NN	O	O
growth	NN	O	O
and	NN	O	O
subsequent	NN	O	O
transformation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
an	NN	O	O
acute	NN	O	O
T-cell	NN	O	O
malignancy	NN	O	O
termed	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
.	NN	O	O

A	NN	O	O
characteristic	NN	O	O
of	NN	O	O
HTLV-I-transformed	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
the	NN	O	O
constitutive	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	O
factors	NN	O	O
,	NN	O	O
which	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
these	NN	O	O
transformed	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
known	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
T-cell	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
interleukin	NN	O	I-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
,	NN	O	O
it	NN	O	O
is	NN	O	O
unclear	NN	O	O
how	NN	O	O
they	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
-independent	NN	O	O
growth	NN	O	O
of	NN	O	O
HTLV-I-transformed	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
certain	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
members	NN	O	I-protein
,	NN	O	O
predominantly	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
interact	NN	O	O
with	NN	O	O
enhancer	NN	O	O
sequences	NN	O	O
for	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
a	NN	O	O
key	NN	O	O
transcription	NN	O	O
factor	NN	O	O
mediating	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Reporter	NN	O	O
gene	NN	O	O
assays	NN	O	O
reveal	NN	O	O
that	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
to	NN	O	O
the	NN	O	O
STAT5	NN	O	B-protein
site	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
Fc	NN	O	B-DNA
gammaR1	NN	O	I-DNA
gene	NN	O	I-DNA
leads	NN	O	O
to	NN	O	O
potent	NN	O	O
transactivation	NN	O	O
of	NN	O	O
this	NN	O	O
enhancer	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
to	NN	O	O
the	NN	O	O
Fc	NN	O	B-DNA
gammaR1	NN	O	I-DNA
STAT	NN	O	I-DNA
site	NN	O	I-DNA
also	NN	O	O
occurs	NN	O	O
in	NN	O	O
human	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
immortalized	NN	O	O
with	NN	O	O
HTLV-I	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
enhanced	NN	O	O
levels	NN	O	O
of	NN	O	O
proliferation	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
raise	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
may	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
growth	NN	O	O
control	NN	O	O
of	NN	O	O
HTLV-I-transformed	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
regulating	NN	O	O
genes	NN	O	O
driven	NN	O	O
by	NN	O	O
both	NN	O	O
kappaB	NN	O	B-DNA
and	NN	O	O
certain	NN	O	O
STAT	NN	O	B-DNA
enhancers	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Activation-dependent	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
Fas	NN	O	I-DNA
promoter	NN	O	I-DNA
requires	NN	O	O
NF-kappaB	NN	O	B-protein
p50-p65	NN	O	I-protein
recruitment	NN	O	O
.	NN	O	O

Fas	NN	O	B-protein
(	NN	O	O
CD95	NN	O	B-protein
)	NN	O	O
and	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
CD95L	NN	O	B-protein
)	NN	O	O
are	NN	O	O
an	NN	O	O
interacting	NN	O	O
receptor-ligand	NN	O	O
pair	NN	O	O
required	NN	O	O
for	NN	O	O
immune	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

Lymphocyte	NN	O	O
activation	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
the	NN	O	O
acquisition	NN	O	O
of	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
FasL	NN	O	B-protein
-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Although	NN	O	O
Fas	NN	O	O
upregulation	NN	O	O
is	NN	O	O
central	NN	O	O
to	NN	O	O
the	NN	O	O
preservation	NN	O	O
of	NN	O	O
immunologic	NN	O	O
tolerance	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
molecular	NN	O	O
machinery	NN	O	O
underlying	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
events	NN	O	O
involved	NN	O	O
in	NN	O	O
activation-induced	NN	O	O
Fas	NN	O	B-protein
upregulation	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
mRNA	NN	O	O
accumulation	NN	O	O
,	NN	O	O
fas	NN	O	O
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
treated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
(	NN	O	O
P/I	NN	O	O
)	NN	O	O
,	NN	O	O
pharmacological	NN	O	O
mimics	NN	O	O
of	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Although	NN	O	O
resting	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
express	NN	O	O
Fas	NN	O	B-protein
,	NN	O	O
Fas	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
induced	NN	O	O
approximately	NN	O	O
10-fold	NN	O	O
in	NN	O	O
2	NN	O	O
h	NN	O	O
upon	NN	O	O
P/I	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Using	NN	O	O
sequential	NN	O	B-DNA
deletion	NN	O	I-DNA
mutants	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
fas	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
47-bp	NN	O	B-DNA
sequence	NN	O	I-DNA
(	NN	O	O
positions	NN	O	B-DNA
-306	NN	O	I-DNA
to	NN	O	I-DNA
-260	NN	O	I-DNA
relative	NN	O	O
to	NN	O	O
the	NN	O	O
ATG	NN	O	O
)	NN	O	O
required	NN	O	O
for	NN	O	O
activation-driven	NN	O	O
fas	NN	O	O
upregulation	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
previously	NN	O	O
unrecognized	NN	O	B-DNA
composite	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
for	NN	O	O
both	NN	O	O
the	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
at	NN	O	O
positions	NN	O	B-DNA
-295	NN	O	I-DNA
to	NN	O	I-DNA
-286	NN	O	I-DNA
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
and	NN	O	O
supershift	NN	O	O
analyses	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
documented	NN	O	O
constitutive	NN	O	O
binding	NN	O	O
of	NN	O	O
Sp1	NN	O	B-protein
in	NN	O	O
unactivated	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
and	NN	O	O
inducible	NN	O	O
binding	NN	O	O
of	NN	O	O
p50-p65	NN	O	B-protein
NF-kappaB	NN	O	I-protein
heterodimers	NN	O	I-protein
after	NN	O	O
P/I	NN	O	O
activation	NN	O	O
.	NN	O	O

Sp1	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	O
factor	NN	O	O
binding	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
mutually	NN	O	O
exclusive	NN	O	O
by	NN	O	O
EMSA	NN	O	O
displacement	NN	O	O
studies	NN	O	O
with	NN	O	O
purified	NN	O	O
recombinant	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
recombinant	NN	O	O
p50	NN	O	B-protein
.	NN	O	O

The	NN	O	O
functional	NN	O	O
contribution	NN	O	O
of	NN	O	O
the	NN	O	O
kappaB-Sp1	NN	O	B-DNA
composite	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
P/I-inducible	NN	O	B-DNA
fas	NN	O	I-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
was	NN	O	O
verified	NN	O	O
by	NN	O	O
using	NN	O	O
kappaB-	NN	O	O
Sp1	NN	O	B-protein
concatamers	NN	O	O
(	NN	O	O
-295	NN	O	B-DNA
to	NN	O	I-DNA
-286	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
a	NN	O	O
thymidine	NN	O	B-DNA
kinase	NN	O	I-DNA
promoter-driven	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
and	NN	O	O
native	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
overexpressing	NN	O	O
IkappaB-alpha	NN	O	B-protein
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
critical	NN	O	O
guanine	NN	O	O
nucleotides	NN	O	O
in	NN	O	O
the	NN	O	O
kappaB-Sp1	NN	O	B-DNA
element	NN	O	I-DNA
documented	NN	O	O
the	NN	O	O
essential	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
site	NN	O	O
in	NN	O	O
activation-dependent	NN	O	O
fas	NN	O	O
promoter	NN	O	O
induction	NN	O	O
.	NN	O	O

-DOCSTART-	O

Evidence	NN	O	O
for	NN	O	O
repression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
activation	NN	O	O
in	NN	O	O
anergic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
clonal	NN	O	O
anergy	NN	O	O
in	NN	O	O
a	NN	O	O
T	NN	O	O
cell	NN	O	O
inhibits	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
because	NN	O	O
of	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
a	NN	O	O
proximal	NN	O	O
signal	NN	O	O
transduction	NN	O	O
defect	NN	O	O
.	NN	O	O

Fusion	NN	O	O
of	NN	O	O
anergic	NN	O	B-cell_type
murine	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
human	NN	O	B-cell_type
Jurkat	NN	O	I-cell_type
T	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
formation	NN	O	O
of	NN	O	O
heterokaryons	NN	O	B-cell_line
failed	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
complementation	NN	O	O
of	NN	O	O
this	NN	O	O
signaling	NN	O	O
defect	NN	O	O
and	NN	O	O
restoration	NN	O	O
of	NN	O	O
murine	NN	O	B-RNA
IL-2	NN	O	I-RNA
mRNA	NN	O	I-RNA
inducibility	NN	O	O
.	NN	O	O

Instead	NN	O	O
,	NN	O	O
signal	NN	O	O
transduction	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
became	NN	O	O
disrupted	NN	O	O
.	NN	O	O

Heterokaryons	NN	O	B-cell_line
formed	NN	O	O
by	NN	O	O
the	NN	O	O
fusion	NN	O	O
of	NN	O	O
anergic	NN	O	B-cell_type
murine	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
normal	NN	O	B-cell_type
murine	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
also	NN	O	O
failed	NN	O	O
to	NN	O	O
accumulate	NN	O	O
intracellular	NN	O	B-protein
IL-2	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
stimulation	NN	O	O
either	NN	O	O
with	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
CD3	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
mAbs	NN	O	I-protein
or	NN	O	O
with	NN	O	O
ionomycin	NN	O	O
plus	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
-activating	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
argue	NN	O	O
against	NN	O	O
a	NN	O	O
loss-of-function	NN	O	O
signaling	NN	O	O
defect	NN	O	O
as	NN	O	O
the	NN	O	O
sole	NN	O	O
basis	NN	O	O
for	NN	O	O
clonal	NN	O	O
anergy	NN	O	O
induction	NN	O	O
and	NN	O	O
document	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
dominant-acting	NN	O	B-protein
repressor	NN	O	I-protein
molecule	NN	O	I-protein
that	NN	O	O
inhibits	NN	O	O
signal	NN	O	O
transduction	NN	O	O
to	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
within	NN	O	O
viable	NN	O	O
anergic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Comparison	NN	O	O
of	NN	O	O
HTLV-I	NN	O	O
basal	NN	O	O
transcription	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
CREB/ATF-1/CREM	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

HTLV-I	NN	O	O
is	NN	O	O
the	NN	O	O
etiologic	NN	O	O
agent	NN	O	O
of	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia/lymphoma	NN	O	O
and	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
tropical	NN	O	O
spastic	NN	O	O
paraparesis/HTLV-I-associated	NN	O	O
myelopathy	NN	O	O
.	NN	O	O

Following	NN	O	O
integration	NN	O	O
into	NN	O	O
the	NN	O	O
host	NN	O	O
cell	NN	O	O
genome	NN	O	O
,	NN	O	O
HTLV-I	NN	O	O
replication	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
both	NN	O	O
host	NN	O	O
and	NN	O	O
viral	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
transcription	NN	O	O
.	NN	O	O

Low	NN	O	O
levels	NN	O	O
of	NN	O	O
viral	NN	O	O
transcription	NN	O	O
(	NN	O	O
basal	NN	O	O
transcription	NN	O	O
)	NN	O	O
occur	NN	O	O
before	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
virally	NN	O	B-protein
encoded	NN	O	I-protein
Tax	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
Tax	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
)	NN	O	O
.	NN	O	O

Members	NN	O	O
of	NN	O	O
the	NN	O	O
cyclic	NN	O	B-protein
adenosine	NN	O	I-protein
monophosphate	NN	O	I-protein
(	NN	O	I-protein
cAMP	NN	O	I-protein
)	NN	O	I-protein
response	NN	O	I-protein
element	NN	O	I-protein
binding	NN	O	I-protein
(	NN	O	I-protein
CREB	NN	O	I-protein
)	NN	O	I-protein
/activating	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
ATF-1	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
bind	NN	O	O
three	NN	O	O
21-bp	NN	O	B-DNA
repeats	NN	O	I-DNA
(	NN	O	O
Tax-responsive	NN	O	B-DNA
element-1	NN	O	I-DNA
,	NN	O	O
or	NN	O	O
TRE-1	NN	O	B-DNA
)	NN	O	O
within	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
basal	NN	O	O
and	NN	O	O
Tax-mediated	NN	O	O
transcription	NN	O	O
.	NN	O	O

Using	NN	O	O
mitogen	NN	O	O
stimulated	NN	O	O
and	NN	O	O
quiescent	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
compared	NN	O	O
differences	NN	O	O
in	NN	O	O
basal	NN	O	O
transcription	NN	O	O
and	NN	O	O
amounts	NN	O	O
and	NN	O	O
binding	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
with	NN	O	O
TRE-1	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
amounts	NN	O	O
of	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
phosphorylated	NN	O	B-protein
CREB	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
P-CREB	NN	O	B-protein
)	NN	O	O
differ	NN	O	O
between	NN	O	O
activated	NN	O	O
PBMC	NN	O	B-cell_type
and	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Following	NN	O	O
stimulation	NN	O	O
,	NN	O	O
P-CREB	NN	O	B-protein
levels	NN	O	O
remain	NN	O	O
elevated	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
for	NN	O	O
up	NN	O	O
to	NN	O	O
24	NN	O	O
hours	NN	O	O
whereas	NN	O	O
CREB	NN	O	O
is	NN	O	O
dephosphorylated	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
within	NN	O	O
4	NN	O	O
hours	NN	O	O
following	NN	O	O
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
differences	NN	O	O
in	NN	O	O
P-CREB	NN	O	B-protein
levels	NN	O	O
between	NN	O	O
PBMC	NN	O	B-cell_type
and	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
directly	NN	O	O
correlated	NN	O	O
with	NN	O	O
basal	NN	O	O
transcription	NN	O	O
of	NN	O	O
HTLV-I	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
that	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
band	NN	O	O
migration	NN	O	O
differed	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
PBMC	NN	O	B-cell_type
differentially	NN	O	O
regulate	NN	O	O
basal	NN	O	O
HTLV-I	NN	O	O
transcription	NN	O	O
compared	NN	O	O
with	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
this	NN	O	O
differential	NN	O	O
regulation	NN	O	O
is	NN	O	O
due	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
to	NN	O	O
differential	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
binding	NN	O	O
of	NN	O	O
CREB/ATF-1	NN	O	B-protein
to	NN	O	O
TRE-1	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
HTLV-I	NN	O	O
promoter	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
utility	NN	O	O
of	NN	O	O
using	NN	O	O
primary	NN	O	O
lymphocyte	NN	O	O
models	NN	O	O
to	NN	O	O
study	NN	O	O
HTLV-I	NN	O	O
transcription	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
cell	NN	O	O
signaling	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
activated	NN	O	O
PBMC	NN	O	B-cell_type
maintain	NN	O	O
elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
P-CREB	NN	O	B-protein
,	NN	O	O
which	NN	O	O
promote	NN	O	O
basal	NN	O	O
HTLV-I	NN	O	O
transcription	NN	O	O
and	NN	O	O
enhance	NN	O	O
viral	NN	O	O
persistence	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Anoxia/reoxygenation-induced	NN	O	O
tolerance	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
polymorphonuclear	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
cultured	NN	O	B-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
-mediated	NN	O	O
phenomenon	NN	O	O
.	NN	O	O

Exposing	NN	O	O
human	NN	O	B-cell_line
umbilical	NN	O	I-cell_line
vein	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
HUVECs	NN	O	B-cell_line
)	NN	O	O
to	NN	O	O
anoxia/reoxygenation	NN	O	O
(	NN	O	O
A/R	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
polymorphonuclear	NN	O	O
leukocyte	NN	O	O
(	NN	O	O
PMN	NN	O	B-cell_type
)	NN	O	O
adhesion	NN	O	O
to	NN	O	O
HUVECs	NN	O	B-cell_line
.	NN	O	O

This	NN	O	O
A/R-induced	NN	O	O
hyperadhesion	NN	O	O
is	NN	O	O
completely	NN	O	O
prevented	NN	O	O
by	NN	O	O
a	NN	O	O
previous	NN	O	O
(	NN	O	O
24	NN	O	O
hours	NN	O	O
earlier	NN	O	O
)	NN	O	O
exposure	NN	O	O
of	NN	O	O
HUVECs	NN	O	B-cell_line
to	NN	O	O
A/R	NN	O	O
.	NN	O	O

This	NN	O	O
phenomenon	NN	O	O
has	NN	O	O
been	NN	O	O
termed	NN	O	O
``	NN	O	O
A/R	NN	O	O
tolerance.	NN	O	O
''	NN	O	O

Exposing	NN	O	O
HUVECs	NN	O	B-cell_line
to	NN	O	O
A/R	NN	O	O
induces	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
in	NN	O	O
HUVEC	NN	O	O
nuclei	NN	O	O
within	NN	O	O
4	NN	O	O
hours	NN	O	O
.	NN	O	O

Interfering	NN	O	O
with	NN	O	O
either	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
(	NN	O	O
proteasome	NN	O	B-protein
inhibitor	NN	O	O
)	NN	O	O
or	NN	O	O
translocation	NN	O	O
(	NN	O	O
double-stranded	NN	O	O
oligonucleotides	NN	O	O
containing	NN	O	O
NF-kappaB	NN	O	B-DNA
binding	NN	O	I-DNA
sequence	NN	O	I-DNA
)	NN	O	O
prevents	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
A/R	NN	O	O
tolerance	NN	O	O
(	NN	O	O
ie	NN	O	O
,	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
A/R-induced	NN	O	O
PMN	NN	O	O
adhesion	NN	O	O
to	NN	O	O
HUVECs	NN	O	B-cell_line
is	NN	O	O
the	NN	O	O
same	NN	O	O
after	NN	O	O
the	NN	O	O
first	NN	O	O
and	NN	O	O
second	NN	O	O
A/R	NN	O	O
challenges	NN	O	O
)	NN	O	O
.	NN	O	O

NO	NN	O	O
production	NN	O	O
by	NN	O	O
HUVECs	NN	O	B-cell_line
is	NN	O	O
increased	NN	O	O
after	NN	O	O
the	NN	O	O
second	NN	O	O
A/R	NN	O	O
challenge	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
after	NN	O	O
the	NN	O	O
first	NN	O	O
A/R	NN	O	O
challenge	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
NO	NN	O	B-protein
synthase	NN	O	I-protein
(	NN	O	O
NOS	NN	O	B-protein
)	NN	O	O
during	NN	O	O
the	NN	O	O
second	NN	O	O
A/R	NN	O	O
challenge	NN	O	O
prevents	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
A/R	NN	O	O
tolerance	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
PMN	NN	O	O
adhesion	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
while	NN	O	O
HUVECs	NN	O	B-cell_line
contained	NN	O	O
endothelial	NN	O	O
NOS	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
no	NN	O	O
inducible	NN	O	O
NOS	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
either	NN	O	O
tolerant	NN	O	O
or	NN	O	O
nontolerant	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Further	NN	O	O
studies	NN	O	O
indicated	NN	O	O
that	NN	O	O
inhibition	NN	O	O
of	NN	O	O
GTP-cyclohydrolase	NN	O	B-protein
I	NN	O	I-protein
(	NN	O	O
an	NN	O	O
enzyme	NN	O	O
involved	NN	O	O
in	NN	O	O
de	NN	O	O
novo	NN	O	O
synthesis	NN	O	O
of	NN	O	O
an	NN	O	O
important	NN	O	O
cofactor	NN	O	O
for	NN	O	O
NOS	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
tetrahydrobiopterin	NN	O	O
)	NN	O	O
prevented	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
NO	NN	O	O
in	NN	O	O
A/R-tolerant	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Extracellular	NN	O	O
generation	NN	O	O
of	NN	O	O
NO	NN	O	O
(	NN	O	O
NO	NN	O	O
donor	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
effect	NN	O	O
the	NN	O	O
hyperadhesion	NN	O	O
response	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
initial	NN	O	O
A/R	NN	O	O
challenge	NN	O	O
.	NN	O	O

A/R	NN	O	O
also	NN	O	O
induced	NN	O	O
an	NN	O	O
oxidant	NN	O	O
stress	NN	O	O
in	NN	O	O
naive	NN	O	O
HUVECs	NN	O	B-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
A/R-tolerant	NN	O	B-cell_line
HUVECs	NN	O	I-cell_line
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
NOS	NN	O	B-protein
during	NN	O	O
the	NN	O	O
second	NN	O	O
A/R	NN	O	O
insult	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
an	NN	O	O
oxidant	NN	O	O
stress	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
observed	NN	O	O
after	NN	O	O
the	NN	O	O
first	NN	O	O
A/R	NN	O	O
challenge	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
the	NN	O	O
findings	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
A/R	NN	O	O
tolerance	NN	O	O
(	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
PMN	NN	O	B-cell_type
adhesion	NN	O	O
)	NN	O	O
,	NN	O	O
perhaps	NN	O	O
by	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
GTP-cyclohydrolase	NN	O	B-protein
.	NN	O	O

The	NN	O	O
increased	NN	O	O
NO	NN	O	O
production	NN	O	O
during	NN	O	O
the	NN	O	O
second	NN	O	O
A/R	NN	O	O
insult	NN	O	O
reduces	NN	O	O
PMN	NN	O	O
adhesion	NN	O	O
most	NN	O	O
likely	NN	O	O
by	NN	O	O
reducing	NN	O	O
the	NN	O	O
intracellular	NN	O	O
oxidant	NN	O	O
stress	NN	O	O
induced	NN	O	O
by	NN	O	O
A/R	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mildly	NN	O	O
oxidized	NN	O	O
low-density	NN	O	B-protein
lipoproteins	NN	O	I-protein
decrease	NN	O	O
early	NN	O	O
production	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
.	NN	O	O

Activated	NN	O	O
T-lymphocytes	NN	O	O
are	NN	O	O
found	NN	O	O
early	NN	O	O
in	NN	O	O
atherosclerosis	NN	O	O
lesions	NN	O	O
,	NN	O	O
but	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
their	NN	O	O
role	NN	O	O
.	NN	O	O

Oxidized	NN	O	O
low-density	NN	O	B-protein
lipoproteins	NN	O	I-protein
(	NN	O	O
oxLDLs	NN	O	B-protein
)	NN	O	O
are	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
the	NN	O	O
lesions	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
have	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
oxLDLs	NN	O	B-protein
inhibit	NN	O	O
not	NN	O	O
only	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2-receptor	NN	O	I-protein
expression	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
in	NN	O	O
vitro-activated	NN	O	O
T-lymphocytes	NN	O	O
but	NN	O	O
also	NN	O	O
their	NN	O	O
proliferation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
oxLDLs	NN	O	B-protein
on	NN	O	O
blast	NN	O	O
differentiation	NN	O	O
,	NN	O	O
on	NN	O	O
IL-2	NN	O	B-protein
synthesis	NN	O	O
and	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
system	NN	O	O
in	NN	O	O
activated	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

Mildly	NN	O	O
oxLDLs	NN	O	B-protein
(	NN	O	O
50	NN	O	O
and	NN	O	O
100	NN	O	O
microgram/ml	NN	O	O
)	NN	O	O
decreased	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
lymphoblasts	NN	O	B-cell_type
and	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
IL-2	NN	O	O
concentration	NN	O	O
in	NN	O	O
the	NN	O	O
culture	NN	O	O
supernatants	NN	O	O
after	NN	O	O
activation	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
by	NN	O	O
phytohaemagglutinin	NN	O	B-protein
and	NN	O	O
PMA+ionomycin	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
(	NN	O	O
+	NN	O	O
)	NN	O	O
T-lymphocyte	NN	O	O
cytoplasm	NN	O	O
as	NN	O	O
early	NN	O	O
as	NN	O	O
4	NN	O	O
h	NN	O	O
after	NN	O	O
activation	NN	O	O
by	NN	O	O
PMA+ionomycin	NN	O	O
.	NN	O	O

The	NN	O	O
study	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
showed	NN	O	O
that	NN	O	O
oxLDLs	NN	O	B-protein
led	NN	O	O
to	NN	O	O
a	NN	O	O
decrease	NN	O	O
of	NN	O	O
activation-induced	NN	O	B-protein
p65/p50	NN	O	I-protein
NF-kappaB	NN	O	B-protein
heterodimer	NN	O	O
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
constitutive	NN	O	O
nuclear	NN	O	O
form	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
dimer	NN	O	I-protein
was	NN	O	O
unchanged	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
correlated	NN	O	O
with	NN	O	O
an	NN	O	O
unchanged	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
active	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
cytosolic	NN	O	B-protein
inhibitor	NN	O	I-protein
protein	NN	O	I-protein
IkappaB-alpha	NN	O	B-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
effect	NN	O	O
of	NN	O	O
oxLDLs	NN	O	B-protein
might	NN	O	O
operate	NN	O	O
via	NN	O	O
a	NN	O	O
dysregulation	NN	O	O
of	NN	O	O
the	NN	O	O
T-lymphocyte	NN	O	B-cell_type
activation	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
B29	NN	O	B-DNA
(	NN	O	I-DNA
immunoglobulin	NN	O	I-DNA
beta-chain	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
genetic	NN	O	O
target	NN	O	O
for	NN	O	O
early	NN	O	B-protein
B-cell	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Early	NN	O	B-protein
B-cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EBF	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
suggested	NN	O	O
as	NN	O	O
essential	NN	O	O
for	NN	O	O
early	NN	O	O
B-lymphocyte	NN	O	B-cell_type
development	NN	O	O
by	NN	O	O
findings	NN	O	O
in	NN	O	O
mice	NN	O	O
where	NN	O	O
the	NN	O	O
coding	NN	O	O
gene	NN	O	O
has	NN	O	O
been	NN	O	O
inactivated	NN	O	O
by	NN	O	O
homologous	NN	O	O
disruption	NN	O	O
.	NN	O	O

This	NN	O	O
makes	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
genetic	NN	O	O
targets	NN	O	O
for	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
pertinent	NN	O	O
for	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
early	NN	O	O
B-cell	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
lack	NN	O	O
of	NN	O	O
B29	NN	O	B-RNA
transcripts	NN	O	I-RNA
,	NN	O	O
coding	NN	O	O
for	NN	O	O
the	NN	O	O
beta	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
B-cell	NN	O	O
receptor	NN	O	O
complex	NN	O	O
,	NN	O	O
in	NN	O	O
pro-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
EBF	NN	O	B-protein
-deficient	NN	O	O
mice	NN	O	O
suggested	NN	O	O
that	NN	O	O
B29	NN	O	O
might	NN	O	O
be	NN	O	O
a	NN	O	O
genetic	NN	O	O
target	NN	O	O
for	NN	O	O
EBF	NN	O	B-protein
.	NN	O	O

We	NN	O	O
here	NN	O	O
present	NN	O	O
data	NN	O	O
suggesting	NN	O	O
that	NN	O	O
EBF	NN	O	B-protein
interacts	NN	O	O
with	NN	O	O
three	NN	O	O
independent	NN	O	O
sites	NN	O	O
within	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
B29	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
EBF	NN	O	B-protein
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
activated	NN	O	O
a	NN	O	O
B29	NN	O	B-DNA
promoter-controlled	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
13-fold	NN	O	O
and	NN	O	O
induced	NN	O	O
a	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
from	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
B29	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
EBF	NN	O	B-protein
binding	NN	O	O
sites	NN	O	O
diminished	NN	O	O
B29	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
pre-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
while	NN	O	O
the	NN	O	O
same	NN	O	O
mutations	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
as	NN	O	O
striking	NN	O	O
an	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
promoter	NN	O	O
function	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
of	NN	O	O
later	NN	O	O
differentiation	NN	O	O
stages	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
B29	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
genetic	NN	O	O
target	NN	O	O
for	NN	O	O
EBF	NN	O	B-protein
in	NN	O	O
early	NN	O	O
B-cell	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Delta-opioid	NN	O	O
receptors	NN	O	O
expressed	NN	O	O
by	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
enhance	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
increasing	NN	O	O
AP-1	NN	O	B-protein
complexes	NN	O	I-protein
and	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT/AP-1-binding	NN	O	B-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Recent	NN	O	O
molecular	NN	O	O
evidence	NN	O	O
points	NN	O	O
to	NN	O	O
transient	NN	O	O
and/or	NN	O	O
stage-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
delta-	NN	O	B-protein
and	NN	O	I-protein
kappa-opioid	NN	O	I-protein
receptors	NN	O	I-protein
by	NN	O	O
thymic	NN	O	B-cell_type
and	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Since	NN	O	O
medical	NN	O	O
treatments	NN	O	O
or	NN	O	O
stress	NN	O	O
commonly	NN	O	O
increase	NN	O	O
opioid	NN	O	O
levels	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
important	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
opioids	NN	O	O
affect	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
functions	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
created	NN	O	O
and	NN	O	O
studied	NN	O	O
a	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
expressing	NN	O	O
the	NN	O	O
cloned	NN	O	B-protein
delta-opioid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
DOR1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

DOR1	NN	O	B-protein
ligation	NN	O	O
by	NN	O	O
a	NN	O	O
specific	NN	O	O
DOR1	NN	O	B-protein
agonist	NN	O	O
,	NN	O	O
deltorphin	NN	O	O
,	NN	O	O
augmented	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
synergizing	NN	O	O
with	NN	O	O
signals	NN	O	O
from	NN	O	O
TCR-CD3	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

Reporter	NN	O	O
gene	NN	O	O
constructs	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
map	NN	O	O
this	NN	O	O
effect	NN	O	O
of	NN	O	O
deltorphin	NN	O	O
to	NN	O	O
the	NN	O	O
AP-1-	NN	O	B-DNA
and	NN	O	I-DNA
NF-AT/AP-1-binding	NN	O	I-DNA
sites	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Although	NN	O	O
DOR1	NN	O	B-protein
signaling	NN	O	O
increased	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
,	NN	O	O
deltorphin	NN	O	O
enhanced	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT/AP-1-binding	NN	O	B-DNA
site	NN	O	I-DNA
via	NN	O	O
a	NN	O	O
mechanism	NN	O	O
independent	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
and	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
elevated	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
.	NN	O	O

Deltorphin	NN	O	O
also	NN	O	O
increased	NN	O	O
accumulation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complexes	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
DOR1	NN	O	B-protein
augments	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
increasing	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
component	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NF-AT/AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
advance	NN	O	O
the	NN	O	O
molecular	NN	O	O
understanding	NN	O	O
of	NN	O	O
opioid	NN	O	O
effects	NN	O	O
on	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
in	NN	O	O
addition	NN	O	O
,	NN	O	O
demonstrate	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
synthesis	NN	O	O
and	NN	O	O
secretion	NN	O	O
by	NN	O	O
the	NN	O	O
novel	NN	O	O
mechanism	NN	O	O
of	NN	O	O
receptor-mediated	NN	O	O
AP-1	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
interleukin-12	NN	O	B-protein
p40	NN	O	B-RNA
transcript	NN	O	I-RNA
by	NN	O	O
CD40	NN	O	B-protein
ligation	NN	O	O
via	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
.	NN	O	O

Interleukin-12	NN	O	B-protein
is	NN	O	O
produced	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
infection	NN	O	O
with	NN	O	O
bacteria	NN	O	O
or	NN	O	O
parasites	NN	O	O
or	NN	O	O
to	NN	O	O
bacterial	NN	O	O
constituents	NN	O	O
such	NN	O	O
as	NN	O	O
LPS	NN	O	O
in	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
and	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
also	NN	O	O
generated	NN	O	O
by	NN	O	O
the	NN	O	O
interaction	NN	O	O
between	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
CD40-CD40	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
CD40L	NN	O	B-protein
)	NN	O	O
.	NN	O	O

So	NN	O	O
far	NN	O	O
,	NN	O	O
transcriptional	NN	O	O
analyses	NN	O	O
of	NN	O	O
p40	NN	O	B-protein
have	NN	O	O
been	NN	O	O
carried	NN	O	O
out	NN	O	O
only	NN	O	O
using	NN	O	O
bacterial	NN	O	O
constituents	NN	O	O
such	NN	O	O
as	NN	O	O
LPS	NN	O	O
as	NN	O	O
stimuli	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
p40	NN	O	B-protein
by	NN	O	O
CD40	NN	O	B-protein
ligation	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
Daudi	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
acute	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
.	NN	O	O

These	NN	O	O
cells	NN	O	O
,	NN	O	O
stimulated	NN	O	O
by	NN	O	O
an	NN	O	O
agonistic	NN	O	O
monoclonal	NN	O	O
antibody	NN	O	O
against	NN	O	O
CD40	NN	O	B-protein
or	NN	O	O
by	NN	O	O
transfection	NN	O	O
with	NN	O	O
a	NN	O	O
CD40L	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
,	NN	O	O
secreted	NN	O	O
p40	NN	O	B-protein
and	NN	O	O
showed	NN	O	O
enhanced	NN	O	O
p40	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
p40	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
identified	NN	O	O
two	NN	O	O
potential	NN	O	B-DNA
nuclear	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
NF	NN	O	I-DNA
)	NN	O	I-DNA
-kappaB	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
conserved	NN	O	O
between	NN	O	O
mouse	NN	O	O
and	NN	O	O
human	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
potential	NN	O	O
NF-kappaB	NN	O	B-DNA
binding	NN	O	I-DNA
sequence	NN	O	I-DNA
which	NN	O	O
is	NN	O	O
located	NN	O	O
around	NN	O	O
120	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
murine	NN	O	B-DNA
and	NN	O	I-DNA
human	NN	O	I-DNA
p40	NN	O	I-DNA
genes	NN	O	I-DNA
formed	NN	O	O
an	NN	O	O
NF-kappaB	NN	O	B-protein
complex	NN	O	I-protein
with	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
from	NN	O	O
Daudi	NN	O	B-cell_line
cells	NN	O	I-cell_line
stimulated	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
ligation	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
transfection	NN	O	O
of	NN	O	O
Daudi	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
polymerized	NN	O	O
NF-kappaB	NN	O	B-DNA
binding	NN	O	I-DNA
sequence	NN	O	I-DNA
ligated	NN	O	O
to	NN	O	O
a	NN	O	O
thymidine	NN	O	B-DNA
kinase/chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
reporter	NN	O	I-DNA
plasmid	NN	O	I-DNA
greatly	NN	O	O
induced	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
but	NN	O	O
transfection	NN	O	O
with	NN	O	O
the	NN	O	O
polymerized	NN	O	B-DNA
mutated	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
binding	NN	O	I-DNA
sequence	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
located	NN	O	O
around	NN	O	O
120	NN	O	O
bp	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
murine	NN	O	B-DNA
and	NN	O	I-DNA
human	NN	O	I-DNA
p40	NN	O	I-DNA
promoter	NN	O	I-DNA
regions	NN	O	I-DNA
could	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
p40	NN	O	B-protein
induction	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
ligation	NN	O	O
via	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

High	NN	O	O
frequency	NN	O	O
of	NN	O	O
germ-line	NN	O	O
BRCA2	NN	O	B-DNA
mutations	NN	O	O
among	NN	O	O
Hungarian	NN	O	O
male	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
without	NN	O	O
family	NN	O	O
history	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
BRCA1	NN	O	B-DNA
and	NN	O	O
BRCA2	NN	O	B-DNA
mutations	NN	O	O
to	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
male	NN	O	O
breast	NN	O	O
cancer	NN	O	O
in	NN	O	O
Hungary	NN	O	O
,	NN	O	O
the	NN	O	O
country	NN	O	O
with	NN	O	O
the	NN	O	O
highest	NN	O	O
male	NN	O	O
breast	NN	O	O
cancer	NN	O	O
mortality	NN	O	O
rates	NN	O	O
in	NN	O	O
continental	NN	O	O
Europe	NN	O	O
,	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
18	NN	O	O
male	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
and	NN	O	O
three	NN	O	O
patients	NN	O	O
with	NN	O	O
gynecomastia	NN	O	O
was	NN	O	O
analyzed	NN	O	O
for	NN	O	O
germ-line	NN	O	O
mutations	NN	O	O
in	NN	O	O
both	NN	O	O
BRCA1	NN	O	B-DNA
and	NN	O	O
BRCA2	NN	O	B-DNA
.	NN	O	O

Although	NN	O	O
no	NN	O	O
germ-line	NN	O	O
BRCA1	NN	O	B-DNA
mutation	NN	O	O
was	NN	O	O
observed	NN	O	O
,	NN	O	O
6	NN	O	O
of	NN	O	O
the	NN	O	O
18	NN	O	O
male	NN	O	O
breast	NN	O	O
cancer	NN	O	O
cases	NN	O	O
(	NN	O	O
33	NN	O	O
%	NN	O	O
)	NN	O	O
carried	NN	O	O
truncating	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
BRCA2	NN	O	B-DNA
gene	NN	O	O
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
none	NN	O	O
of	NN	O	O
them	NN	O	O
reported	NN	O	O
a	NN	O	O
family	NN	O	O
history	NN	O	O
for	NN	O	O
breast/ovarian	NN	O	O
cancer	NN	O	O
.	NN	O	O

Four	NN	O	O
of	NN	O	O
six	NN	O	O
truncating	NN	O	O
mutations	NN	O	O
were	NN	O	O
novel	NN	O	O
,	NN	O	O
and	NN	O	O
two	NN	O	O
mutations	NN	O	O
were	NN	O	O
recurrent	NN	O	O
.	NN	O	O

Four	NN	O	O
patients	NN	O	O
(	NN	O	O
22	NN	O	O
%	NN	O	O
)	NN	O	O
had	NN	O	O
a	NN	O	O
family	NN	O	O
history	NN	O	O
of	NN	O	O
breast/ovarian	NN	O	O
cancer	NN	O	O
in	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
first-	NN	O	O
or	NN	O	O
second-degree	NN	O	O
relative	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
no	NN	O	O
BRCA2	NN	O	B-DNA
mutation	NN	O	O
was	NN	O	O
identified	NN	O	O
among	NN	O	O
them	NN	O	O
.	NN	O	O

No	NN	O	O
mutation	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
either	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
in	NN	O	O
the	NN	O	O
gynecomastias	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
strong	NN	O	O
genetic	NN	O	O
component	NN	O	O
of	NN	O	O
male	NN	O	O
breast	NN	O	O
cancer	NN	O	O
in	NN	O	O
Hungary	NN	O	O
.	NN	O	O

-DOCSTART-	O

T-cell	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GATA-3	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
a	NN	O	O
non-lineage-specific	NN	O	B-DNA
silencer	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
GATA-3	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	B-cell_line
lineage	NN	O	I-cell_line
and	NN	O	O
Th2	NN	O	B-DNA
cytokine	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
CD4	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
mapped	NN	O	O
the	NN	O	O
DNase-I-hypersensitive	NN	O	B-DNA
(	NN	O	I-DNA
HS	NN	O	I-DNA
)	NN	O	I-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GATA-3	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
T-cells	NN	O	B-cell_type
and	NN	O	O
non-T-cells	NN	O	B-cell_type
and	NN	O	O
studied	NN	O	O
their	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
.	NN	O	O

HS	NN	O	B-DNA
I-III	NN	O	I-DNA
,	NN	O	O
located	NN	O	O
5	NN	O	O
'	NN	O	O
from	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
and	NN	O	I-cell_type
non-hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
HS	NN	O	B-DNA
IV-VII	NN	O	I-DNA
,	NN	O	O
located	NN	O	O
3	NN	O	O
'	NN	O	O
from	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
were	NN	O	O
exclusively	NN	O	O
observed	NN	O	O
in	NN	O	O
T-cells	NN	O	B-cell_type
.	NN	O	O

Among	NN	O	O
these	NN	O	O
hypersensitive	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
two	NN	O	O
transcriptional	NN	O	B-DNA
control	NN	O	I-DNA
elements	NN	O	I-DNA
were	NN	O	O
found	NN	O	O
,	NN	O	O
one	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	B-DNA
intron	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
GATA-3	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
other	NN	O	O
between	NN	O	B-DNA
8.3	NN	O	I-DNA
and	NN	O	I-DNA
5.9	NN	O	I-DNA
kilobases	NN	O	I-DNA
5	NN	O	O
'	NN	O	O
from	NN	O	O
the	NN	O	O
GATA-3	NN	O	B-DNA
transcriptional	NN	O	I-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
first	NN	O	B-DNA
intron	NN	O	I-DNA
acted	NN	O	O
as	NN	O	O
a	NN	O	O
strong	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
in	NN	O	O
a	NN	O	O
position-dependent	NN	O	O
manner	NN	O	O
and	NN	O	O
with	NN	O	O
no	NN	O	O
cell-type	NN	O	O
specificity	NN	O	O
.	NN	O	O

The	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
could	NN	O	O
confer	NN	O	O
T-cell	NN	O	O
specificity	NN	O	O
to	NN	O	O
the	NN	O	O
GATA-3	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
,	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
revealed	NN	O	O
a	NN	O	O
707-base	NN	O	B-DNA
pair	NN	O	I-DNA
silencer	NN	O	I-DNA
that	NN	O	O
drastically	NN	O	O
inhibited	NN	O	O
GATA-3	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
non-T-cells	NN	O	B-cell_type
.	NN	O	O

Two	NN	O	O
CAGGTG	NN	O	O
E-boxes	NN	O	O
,	NN	O	O
located	NN	O	O
at	NN	O	O
the	NN	O	O
5'-	NN	O	B-DNA
and	NN	O	I-DNA
3'-ends	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
silencer	NN	O	O
,	NN	O	O
were	NN	O	O
necessary	NN	O	O
for	NN	O	O
this	NN	O	O
silencer	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
3'-CAGGTG	NN	O	B-DNA
E-box	NN	O	I-DNA
could	NN	O	O
bind	NN	O	O
USF	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
the	NN	O	O
ubiquitous	NN	O	B-protein
repressor	NN	O	I-protein
ZEB	NN	O	B-protein
,	NN	O	O
or	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
proteins	NN	O	I-protein
E2A	NN	O	B-protein
and	NN	O	O
HEB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
a	NN	O	O
competition	NN	O	O
between	NN	O	O
ZEB	NN	O	B-protein
and	NN	O	O
E2A/HEB	NN	O	B-protein
proteins	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
silencer	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reactive	NN	O	O
oxygen	NN	O	O
intermediate-dependent	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
interleukin-1beta	NN	O	B-protein
requires	NN	O	O
5-lipoxygenase	NN	O	B-protein
or	NN	O	O
NADPH	NN	O	O
oxidase	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
(	NN	O	O
ROIs	NN	O	O
)	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
was	NN	O	O
cell	NN	O	O
specific	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
sources	NN	O	O
for	NN	O	O
ROIs	NN	O	O
in	NN	O	O
various	NN	O	O
cell	NN	O	O
types	NN	O	O
are	NN	O	O
yet	NN	O	O
to	NN	O	O
be	NN	O	O
determined	NN	O	O
and	NN	O	O
might	NN	O	O
include	NN	O	O
5-lipoxygenase	NN	O	B-protein
(	NN	O	O
5-LOX	NN	O	B-protein
)	NN	O	O
and	NN	O	O
NADPH	NN	O	O
oxidase	NN	O	O
.	NN	O	O

5-LOX	NN	O	B-protein
and	NN	O	O
5-LOX	NN	O	B-protein
activating	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
FLAP	NN	O	B-protein
)	NN	O	O
are	NN	O	O
coexpressed	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
or	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
interleukin-1beta	NN	O	B-protein
(	NN	O	O
IL-1beta	NN	O	B-protein
)	NN	O	O
led	NN	O	O
to	NN	O	O
ROI	NN	O	O
production	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
which	NN	O	O
could	NN	O	O
both	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
antioxidants	NN	O	O
or	NN	O	O
FLAP	NN	O	B-protein
inhibitors	NN	O	O
,	NN	O	O
confirming	NN	O	O
that	NN	O	O
5-LOX	NN	O	B-protein
was	NN	O	O
the	NN	O	O
source	NN	O	O
of	NN	O	O
ROIs	NN	O	O
and	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

IL-1beta	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
did	NN	O	O
not	NN	O	O
generate	NN	O	O
any	NN	O	O
ROIs	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
induction	NN	O	O
was	NN	O	O
not	NN	O	O
influenced	NN	O	O
by	NN	O	O
5-LOX	NN	O	B-protein
inhibitors	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
reintroduction	NN	O	O
of	NN	O	O
a	NN	O	O
functional	NN	O	O
5-LOX	NN	O	B-protein
system	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
allowed	NN	O	O
ROI	NN	O	O
production	NN	O	O
and	NN	O	O
5-LOX	NN	O	B-protein
-dependent	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

In	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
IL-1beta	NN	O	B-protein
treatment	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
production	NN	O	O
of	NN	O	O
ROIs	NN	O	O
which	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
5-LOX	NN	O	B-protein
enzyme	NN	O	I-protein
but	NN	O	O
requires	NN	O	O
the	NN	O	O
NADPH	NN	O	O
oxidase	NN	O	O
activity	NN	O	O
.	NN	O	O

This	NN	O	O
pathway	NN	O	O
involves	NN	O	O
the	NN	O	O
Rac1	NN	O	B-protein
and	NN	O	O
Cdc42	NN	O	B-protein
GTPases	NN	O	B-protein
,	NN	O	O
two	NN	O	O
enzymes	NN	O	O
which	NN	O	O
are	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
IL-1beta	NN	O	B-protein
in	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
three	NN	O	O
different	NN	O	O
cell-specific	NN	O	O
pathways	NN	O	O
lead	NN	O	O
to	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
IL-1beta	NN	O	B-protein
:	NN	O	O
a	NN	O	O
pathway	NN	O	O
dependent	NN	O	O
on	NN	O	O
ROI	NN	O	O
production	NN	O	O
by	NN	O	O
5-LOX	NN	O	B-protein
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
an	NN	O	O
ROI-	NN	O	O
and	NN	O	O
5-LOX	NN	O	B-protein
-independent	NN	O	O
pathway	NN	O	O
in	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
a	NN	O	O
pathway	NN	O	O
requiring	NN	O	O
ROI	NN	O	O
production	NN	O	O
by	NN	O	O
NADPH	NN	O	B-protein
oxidase	NN	O	I-protein
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Cutting	NN	O	O
edge	NN	O	O
:	NN	O	O
dominant	NN	O	O
effect	NN	O	O
of	NN	O	O
Ile50Val	NN	O	B-protein
variant	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
IL-4	NN	O	I-protein
receptor	NN	O	I-protein
alpha-chain	NN	O	I-protein
in	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
.	NN	O	O

Two	NN	O	O
variants	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-DNA
alpha-chain	NN	O	I-DNA
(	NN	O	I-DNA
IL-4Ralpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
recently	NN	O	O
identified	NN	O	O
in	NN	O	O
association	NN	O	O
with	NN	O	O
different	NN	O	O
atopic	NN	O	O
disorders	NN	O	O
.	NN	O	O

To	NN	O	O
clarify	NN	O	O
the	NN	O	O
etiological	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
variants	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
of	NN	O	O
transfectants	NN	O	O
with	NN	O	O
four	NN	O	O
kinds	NN	O	O
of	NN	O	O
IL-4Ralpha	NN	O	B-protein
carrying	NN	O	O
either	NN	O	O
Val	NN	O	O
or	NN	O	O
Ile	NN	O	O
at	NN	O	O
50	NN	O	B-protein
and	NN	O	O
either	NN	O	O
Gln	NN	O	O
or	NN	O	O
Arg	NN	O	O
at	NN	O	O
551	NN	O	B-protein
.	NN	O	O

The	NN	O	O
substitution	NN	O	O
of	NN	O	O
Ile	NN	O	O
for	NN	O	O
Val	NN	O	O
augmented	NN	O	O
STAT6	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
proliferation	NN	O	O
,	NN	O	O
and	NN	O	O
transcription	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
Iepsilon	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
that	NN	O	O
of	NN	O	O
Arg	NN	O	O
for	NN	O	O
Gln	NN	O	O
did	NN	O	O
not	NN	O	O
change	NN	O	O
these	NN	O	O
IL-4	NN	O	B-protein
signals	NN	O	O
.	NN	O	O

Arg551	NN	O	O
was	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
atopic	NN	O	O
asthma	NN	O	O
in	NN	O	O
the	NN	O	O
Japanese	NN	O	O
population	NN	O	O
.	NN	O	O

CD23	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
were	NN	O	O
augmented	NN	O	O
in	NN	O	O
Ile50-bearing	NN	O	B-cell_line
PBMC	NN	O	I-cell_line
,	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
bearing	NN	O	O
Val50	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
substitution	NN	O	O
of	NN	O	O
Arg551	NN	O	O
does	NN	O	O
not	NN	O	O
enhance	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
signal	NN	O	O
for	NN	O	O
generation	NN	O	O
of	NN	O	O
germline	NN	O	O
epsilon	NN	O	O
transcript	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
Ile50	NN	O	O
contributes	NN	O	O
to	NN	O	O
enhancement	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
growth-factor-dependent	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
mouse	NN	O	B-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
contains	NN	O	O
progenitors	NN	O	O
endowed	NN	O	O
with	NN	O	O
high	NN	O	O
proliferative	NN	O	O
potential	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor-1	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
CSF-1R	NN	O	B-protein
)	NN	O	O
confers	NN	O	O
long-lasting	NN	O	O
CSF-1	NN	O	B-protein
-dependent	NN	O	O
proliferation	NN	O	O
to	NN	O	O
mouse	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
developed	NN	O	O
mice	NN	O	O
transgenic	NN	O	O
for	NN	O	O
human	NN	O	O
CSF-1R	NN	O	B-protein
because	NN	O	O
mouse	NN	O	O
CSF-1	NN	O	B-protein
can	NN	O	O
not	NN	O	O
activate	NN	O	O
human	NN	O	O
CSF-1R	NN	O	B-protein
.	NN	O	O

Then	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
were	NN	O	O
plated	NN	O	O
onto	NN	O	O
MS-5	NN	O	O
stromal	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
membrane	NN	O	B-protein
form	NN	O	I-protein
of	NN	O	O
human	NN	O	B-protein
CSF-1	NN	O	I-protein
(	NN	O	O
2M-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
combine	NN	O	O
the	NN	O	O
hematopoietic	NN	O	O
supporting	NN	O	O
properties	NN	O	O
of	NN	O	O
stromal	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
proliferative	NN	O	O
effects	NN	O	O
of	NN	O	O
CSF-1	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
derive	NN	O	O
a	NN	O	O
hematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
called	NN	O	O
47.10	NN	O	B-cell_line
,	NN	O	O
that	NN	O	O
grew	NN	O	O
indefinitely	NN	O	O
under	NN	O	O
these	NN	O	O
conditions	NN	O	O
,	NN	O	O
whereas	NN	O	O
no	NN	O	O
cell	NN	O	O
line	NN	O	O
could	NN	O	O
be	NN	O	O
developed	NN	O	O
from	NN	O	O
nontransgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Proliferation	NN	O	O
of	NN	O	O
47.10	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
severely	NN	O	O
affected	NN	O	O
by	NN	O	O
neutralizing	NN	O	O
anti-CSF-1R	NN	O	B-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

Morphologic	NN	O	O
and	NN	O	O
cytofluorometry	NN	O	O
analysis	NN	O	O
established	NN	O	O
that	NN	O	O
most	NN	O	O
47.10	NN	O	B-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
immature	NN	O	B-cell_type
myelomonocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
this	NN	O	O
phenotype	NN	O	O
,	NN	O	O
the	NN	O	O
myeloid	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
PU.1	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
erythroid	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
GATA-1	NN	O	I-protein
,	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
47.10	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
few	NN	O	O
47.10	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
3-5	NN	O	O
%	NN	O	O
)	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
lineage	NN	O	O
specific	NN	O	O
markers	NN	O	O
;	NN	O	O
they	NN	O	O
differentiate	NN	O	O
spontaneously	NN	O	O
to	NN	O	O
lineage-positive	NN	O	B-cell_line
cells	NN	O	I-cell_line
after	NN	O	O
replating	NN	O	O
on	NN	O	O
2M-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
agar	NN	O	O
cultures	NN	O	O
,	NN	O	O
47.10	NN	O	B-cell_line
cells	NN	O	I-cell_line
form	NN	O	O
7-	NN	O	O
and	NN	O	O
14-day	NN	O	O
colonies	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
cocktail	NN	O	O
of	NN	O	O
granulocyte/macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
2.5	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
,	NN	O	O
interleukin-3	NN	O	B-protein
(	NN	O	O
1	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
mouse	NN	O	O
CSF-1	NN	O	B-protein
(	NN	O	O
10	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
.	NN	O	O

Under	NN	O	O
these	NN	O	O
conditions	NN	O	O
,	NN	O	O
about	NN	O	O
0.5	NN	O	O
%	NN	O	O
of	NN	O	O
47.10	NN	O	B-cell_line
cells	NN	O	I-cell_line
formed	NN	O	O
large	NN	O	O
14-day	NN	O	O
colonies	NN	O	O
(	NN	O	O
>	NN	O	O
1	NN	O	O
mm	NN	O	O
)	NN	O	O
composed	NN	O	O
of	NN	O	O
mature	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
granulocytes	NN	O	B-cell_type
,	NN	O	O
reflecting	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
progenitors	NN	O	O
endowed	NN	O	O
with	NN	O	O
high	NN	O	O
proliferative	NN	O	O
potential	NN	O	O
(	NN	O	O
HPP-47.10	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
a	NN	O	O
novel	NN	O	O
continuous	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
presenting	NN	O	O
a	NN	O	O
hierarchical	NN	O	O
structure	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
bone	NN	O	B-cell_line
marrow	NN	O	I-cell_line
progenitor	NN	O	I-cell_line
cell	NN	O	I-cell_line
compartment	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
JunB	NN	O	B-protein
during	NN	O	O
T	NN	O	O
helper	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
restricted	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
helper	NN	O	I-cell_type
1	NN	O	I-cell_type
(	NN	O	I-cell_type
Th1	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
2	NN	O	I-cell_type
(	NN	O	I-cell_type
Th2	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
unclear	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
found	NN	O	O
that	NN	O	O
P1	NN	O	B-DNA
,	NN	O	O
an	NN	O	O
element	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
4	NN	O	I-DNA
(	NN	O	I-DNA
IL-4	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
that	NN	O	O
binds	NN	O	O
AP-1	NN	O	O
,	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
Th2-restricted	NN	O	O
IL-4	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
JunB	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
other	NN	O	O
Jun	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
,	NN	O	O
was	NN	O	O
selectively	NN	O	O
induced	NN	O	O
in	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
not	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
differentiation	NN	O	O
.	NN	O	O

JunB	NN	O	B-protein
has	NN	O	O
previously	NN	O	O
been	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
JunB	NN	O	B-protein
binds	NN	O	O
directly	NN	O	O
to	NN	O	O
the	NN	O	O
P1	NN	O	B-DNA
site	NN	O	I-DNA
and	NN	O	O
synergizes	NN	O	O
with	NN	O	O
c-Maf	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
an	NN	O	O
IL-4	NN	O	B-DNA
luciferase	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

JunB	NN	O	B-protein
-control	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
JunB	NN	O	B-protein
at	NN	O	O
Thr102	NN	O	O
and-	NN	O	O
104	NN	O	O
by	NN	O	O
JNK	NN	O	B-protein
MAP	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

The	NN	O	O
synergy	NN	O	O
between	NN	O	O
c-Maf	NN	O	B-protein
and	NN	O	O
JunB	NN	O	B-protein
can	NN	O	O
be	NN	O	O
attributed	NN	O	O
to	NN	O	O
cooperative	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
facilitated	NN	O	O
by	NN	O	O
JunB	NN	O	B-protein
phosphorylation	NN	O	O
.	NN	O	O

In	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
,	NN	O	O
elevated	NN	O	O
JunB	NN	O	B-protein
levels	NN	O	O
caused	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
Th2	NN	O	B-protein
cytokines	NN	O	I-protein
in	NN	O	O
developing	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

JunB	NN	O	B-protein
also	NN	O	O
upregulated	NN	O	O
IL-4	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
immunization	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
early	NN	O	O
increase	NN	O	O
of	NN	O	O
JunB	NN	O	B-protein
protein	NN	O	O
in	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
provide	NN	O	O
the	NN	O	O
specificity	NN	O	O
for	NN	O	O
c-Maf	NN	O	B-protein
in	NN	O	O
IL-4	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
and	NN	O	O
directs	NN	O	O
thereby	NN	O	O
Th2	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
tax	NN	O	I-protein
protein	NN	O	I-protein
abrogates	NN	O	O
interleukin-2	NN	O	B-protein
dependence	NN	O	O
in	NN	O	O
a	NN	O	O
mouse	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
is	NN	O	O
the	NN	O	O
etiologic	NN	O	O
agent	NN	O	O
of	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Tax	NN	O	B-protein
,	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
crucial	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
,	NN	O	O
since	NN	O	O
it	NN	O	O
transforms	NN	O	O
healthy	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
induces	NN	O	O
tumors	NN	O	O
in	NN	O	O
transgenic	NN	O	O
animals	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
activity	NN	O	O
on	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-cell_line
(	NN	O	I-cell_line
IL-2	NN	O	I-cell_line
)	NN	O	I-cell_line
-dependent	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
CTLL-2	NN	O	B-cell_line
.	NN	O	O

Stable	NN	O	O
expression	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
in	NN	O	O
CTLL-2	NN	O	B-cell_line
transformed	NN	O	O
cell	NN	O	O
growth	NN	O	O
from	NN	O	O
being	NN	O	O
IL-2	NN	O	B-protein
dependent	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
independent	NN	O	O
.	NN	O	O

Tax	NN	O	B-protein
stimulated	NN	O	O
transcription	NN	O	O
through	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
the	NN	O	O
cyclic	NN	O	B-DNA
AMP-responsive	NN	O	I-DNA
element-like	NN	O	I-DNA
sequence	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
HTLV-1	NN	O	O
promoter	NN	O	O
.	NN	O	O

The	NN	O	O
finding	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
mutants	NN	O	O
segregating	NN	O	O
these	NN	O	O
two	NN	O	O
pathways	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
pathway	NN	O	O
was	NN	O	O
essential	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
-independent	NN	O	O
growth	NN	O	O
of	NN	O	O
CTLL-2	NN	O	B-cell_type
cells	NN	O	I-cell_type
while	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
pathway	NN	O	O
was	NN	O	O
unnecessary	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
both	NN	O	O
pathways	NN	O	O
were	NN	O	O
necessary	NN	O	O
for	NN	O	O
another	NN	O	O
transformation-related	NN	O	O
activity	NN	O	O
(	NN	O	O
colony	NN	O	O
formation	NN	O	O
in	NN	O	O
soft	NN	O	O
agar	NN	O	O
)	NN	O	O
of	NN	O	O
CTLL-2/Tax	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
has	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
distinct	NN	O	O
activities	NN	O	O
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
-independent	NN	O	O
T-cell	NN	O	O
transformation	NN	O	O
induced	NN	O	O
by	NN	O	O
HTLV-1	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
its	NN	O	O
well-known	NN	O	O
IL-2	NN	O	B-protein
-dependent	NN	O	O
cell	NN	O	O
transformation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Accessing	NN	O	O
Epstein-Barr	NN	O	O
virus-specific	NN	O	O
T-cell	NN	O	O
memory	NN	O	O
with	NN	O	O
peptide-loaded	NN	O	B-cell_line
dendritic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
conventional	NN	O	O
means	NN	O	O
of	NN	O	O
studying	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
-induced	NN	O	O
cytotoxic	NN	O	B-cell_type
T-lymphocyte	NN	O	I-cell_type
(	NN	O	O
CTL	NN	O	O
)	NN	O	O
memory	NN	O	O
,	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
stimulation	NN	O	O
with	NN	O	O
the	NN	O	O
latently	NN	O	O
infected	NN	O	O
autologous	NN	O	O
lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
LCL	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
has	NN	O	O
important	NN	O	O
limitations	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
it	NN	O	O
gives	NN	O	O
no	NN	O	O
information	NN	O	O
on	NN	O	O
memory	NN	O	O
to	NN	O	O
lytic	NN	O	B-protein
cycle	NN	O	I-protein
antigens	NN	O	I-protein
;	NN	O	O
second	NN	O	O
,	NN	O	O
it	NN	O	O
preferentially	NN	O	O
amplifies	NN	O	O
the	NN	O	O
dominant	NN	O	O
components	NN	O	O
of	NN	O	O
latent	NN	O	O
antigen-specific	NN	O	O
memory	NN	O	O
at	NN	O	O
the	NN	O	O
expense	NN	O	O
of	NN	O	O
key	NN	O	O
subdominant	NN	O	O
reactivities	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
an	NN	O	O
alternative	NN	O	O
approach	NN	O	O
,	NN	O	O
based	NN	O	O
on	NN	O	O
in	NN	O	O
vitro	NN	O	O
stimulation	NN	O	O
with	NN	O	O
epitope	NN	O	B-cell_line
peptide-loaded	NN	O	I-cell_line
dendritic	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
DCs	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
which	NN	O	O
allows	NN	O	O
one	NN	O	O
to	NN	O	O
probe	NN	O	O
the	NN	O	O
CTL	NN	O	O
repertoire	NN	O	O
for	NN	O	O
any	NN	O	O
individual	NN	O	O
reactivity	NN	O	O
of	NN	O	O
choice	NN	O	O
;	NN	O	O
this	NN	O	O
method	NN	O	O
proved	NN	O	O
significantly	NN	O	O
more	NN	O	O
efficient	NN	O	O
than	NN	O	O
stimulation	NN	O	O
with	NN	O	O
peptide	NN	O	O
alone	NN	O	O
.	NN	O	O

Using	NN	O	O
this	NN	O	O
approach	NN	O	O
we	NN	O	O
first	NN	O	O
show	NN	O	O
that	NN	O	O
reactivities	NN	O	O
to	NN	O	O
the	NN	O	O
immunodominant	NN	O	O
and	NN	O	O
subdominant	NN	O	O
lytic	NN	O	B-protein
cycle	NN	O	I-protein
epitopes	NN	O	I-protein
identified	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
primary	NN	O	O
EBV	NN	O	O
infection	NN	O	O
are	NN	O	O
regularly	NN	O	O
detectable	NN	O	O
in	NN	O	O
the	NN	O	O
CTL	NN	O	O
memory	NN	O	O
of	NN	O	O
virus	NN	O	O
carriers	NN	O	O
;	NN	O	O
this	NN	O	O
implies	NN	O	O
that	NN	O	O
in	NN	O	O
such	NN	O	O
carriers	NN	O	O
chronic	NN	O	O
virus	NN	O	O
replication	NN	O	O
remains	NN	O	O
under	NN	O	O
direct	NN	O	O
T-cell	NN	O	O
control	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
show	NN	O	O
that	NN	O	O
subdominant	NN	O	O
latent	NN	O	O
cycle	NN	O	O
reactivities	NN	O	O
to	NN	O	O
epitopes	NN	O	O
in	NN	O	O
the	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
LMP2	NN	O	I-protein
,	NN	O	O
though	NN	O	O
rarely	NN	O	O
undetectable	NN	O	O
in	NN	O	O
LCL	NN	O	B-cell_line
-stimulated	NN	O	O
populations	NN	O	O
,	NN	O	O
can	NN	O	O
be	NN	O	O
reactivated	NN	O	O
by	NN	O	O
DC	NN	O	O
stimulation	NN	O	O
and	NN	O	O
selectively	NN	O	O
expanded	NN	O	O
as	NN	O	O
polyclonal	NN	O	O
CTL	NN	O	B-cell_line
lines	NN	O	I-cell_line
;	NN	O	O
the	NN	O	O
adoptive	NN	O	O
transfer	NN	O	O
of	NN	O	O
such	NN	O	O
preparations	NN	O	O
may	NN	O	O
be	NN	O	O
of	NN	O	O
value	NN	O	O
in	NN	O	O
targeting	NN	O	O
certain	NN	O	O
EBV-positive	NN	O	O
malignancies	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin-2	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
and	NN	O	I-protein
beta	NN	O	I-protein
isoforms	NN	O	I-protein
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
immunosuppressive	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
are	NN	O	O
largely	NN	O	O
due	NN	O	O
to	NN	O	O
transcriptional	NN	O	O
inhibition	NN	O	O
of	NN	O	O
immunologically	NN	O	B-DNA
relevant	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
effects	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
intracellular	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
humans	NN	O	O
,	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-RNA
precursor	NN	O	I-RNA
mRNA	NN	O	I-RNA
gives	NN	O	O
rise	NN	O	O
to	NN	O	O
two	NN	O	O
receptor	NN	O	O
isoforms	NN	O	O
,	NN	O	O
termed	NN	O	O
GRalpha	NN	O	B-protein
and	NN	O	O
GRbeta	NN	O	B-protein
.	NN	O	O

We	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
GRbeta	NN	O	B-protein
could	NN	O	O
antagonize	NN	O	O
GRalpha	NN	O	B-protein
-mediated	NN	O	O
transactivation	NN	O	O
of	NN	O	O
a	NN	O	O
glucocorticoid-responsive	NN	O	B-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
GRE	NN	O	I-DNA
)	NN	O	I-DNA
-driven	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
COS-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
designed	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
GR	NN	O	B-protein
isoforms	NN	O	I-protein
on	NN	O	O
glucocorticoid-mediated	NN	O	O
transrepression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
a	NN	O	O
recently	NN	O	O
developed	NN	O	O
transfection	NN	O	O
technique	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
stimulation	NN	O	O
of	NN	O	O
a	NN	O	O
548	NN	O	B-DNA
bp	NN	O	I-DNA
IL-2	NN	O	I-DNA
promoter-luciferase	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
is	NN	O	O
reversed	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
to	NN	O	O
a	NN	O	O
similar	NN	O	O
extent	NN	O	O
as	NN	O	O
in	NN	O	O
Jurkat	NN	O	O
T	NN	O	O
lymphoma	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
GRalpha	NN	O	B-protein
expression	NN	O	O
vector	NN	O	O
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
a	NN	O	O
GRbeta	NN	O	B-protein
expression	NN	O	O
vector	NN	O	O
alone	NN	O	O
did	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
repression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
GRbeta	NN	O	B-protein
did	NN	O	O
not	NN	O	O
antagonize	NN	O	O
the	NN	O	O
repressive	NN	O	O
effects	NN	O	O
of	NN	O	O
GRalpha	NN	O	B-protein
on	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
GRbeta	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
did	NN	O	O
not	NN	O	O
cause	NN	O	O
significant	NN	O	O
inhibition	NN	O	O
of	NN	O	O
GRalpha	NN	O	B-protein
-induced	NN	O	O
transactivation	NN	O	O
of	NN	O	O
a	NN	O	O
GRE-dependent	NN	O	B-DNA
luciferase	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
either	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
transrepressive	NN	O	O
effect	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
on	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
is	NN	O	O
exclusively	NN	O	O
mediated	NN	O	O
by	NN	O	O
GRalpha	NN	O	B-protein
.	NN	O	O

GRbeta	NN	O	B-protein
can	NN	O	O
neither	NN	O	O
antagonize	NN	O	O
GRalpha	NN	O	B-protein
-mediated	NN	O	O
transactivation	NN	O	O
nor	NN	O	O
transrepression	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
pattern	NN	O	O
of	NN	O	O
GRbeta	NN	O	B-protein
-mediated	NN	O	O
antiglucocorticoid	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Extinction	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
upon	NN	O	O
fusion	NN	O	O
with	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
preceded	NN	O	O
by	NN	O	O
rapid	NN	O	O
nuclear	NN	O	O
depletion	NN	O	O
of	NN	O	O
essential	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
and	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
widespread	NN	O	O
inactivation	NN	O	O
of	NN	O	O
genes	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

When	NN	O	O
immunoglobulin	NN	O	B-cell_type
(	NN	O	I-cell_type
Ig	NN	O	I-cell_type
)	NN	O	I-cell_type
expressing	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
fused	NN	O	O
with	NN	O	O
non-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
Ig	NN	O	O
expression	NN	O	O
is	NN	O	O
rapidly	NN	O	O
suppressed	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
,	NN	O	O
a	NN	O	O
phenomenon	NN	O	O
termed	NN	O	O
extinction	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
fusion	NN	O	O
of	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
either	NN	O	O
diploid	NN	O	B-cell_type
or	NN	O	I-cell_type
tetraploid	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
Daudi	NN	O	B-cell_line
)	NN	O	O
results	NN	O	O
in	NN	O	O
widespread	NN	O	O
extinction	NN	O	O
of	NN	O	O
several	NN	O	O
other	NN	O	O
B	NN	O	B-DNA
cell-encoded	NN	O	I-DNA
genes	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
B	NN	O	B-DNA
cell-expressed	NN	O	I-DNA
genes	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
not	NN	O	O
dependent	NN	O	O
on	NN	O	O
cell-specific	NN	O	O
controls	NN	O	O
is	NN	O	O
unaffected	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
underlying	NN	O	O
Ig	NN	O	B-DNA
gene	NN	O	I-DNA
extinction	NN	O	O
can	NN	O	O
be	NN	O	O
explained	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
a	NN	O	O
lack	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
essential	NN	O	O
for	NN	O	O
Ig	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

These	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
either	NN	O	O
not	NN	O	O
produced	NN	O	O
due	NN	O	O
to	NN	O	O
block	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
their	NN	O	O
respective	NN	O	O
genes	NN	O	O
(	NN	O	O
Oct-2	NN	O	B-DNA
,	NN	O	O
OBF-1	NN	O	B-DNA
,	NN	O	O
PU.1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
or	NN	O	O
are	NN	O	O
rendered	NN	O	O
inactive	NN	O	O
posttranslationally	NN	O	O
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
E47	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

By	NN	O	O
isolating	NN	O	O
Daudi	NN	O	B-cell_line
x	NN	O	I-cell_line
HeLa	NN	O	I-cell_line
heterokaryons	NN	O	I-cell_line
a	NN	O	O
few	NN	O	O
hours	NN	O	O
after	NN	O	O
fusion	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
initial	NN	O	O
fate	NN	O	O
of	NN	O	O
two	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
Ig	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
,	NN	O	O
Oct-2	NN	O	B-DNA
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

This	NN	O	O
report	NN	O	O
provides	NN	O	O
the	NN	O	O
first	NN	O	O
demonstration	NN	O	O
that	NN	O	O
upon	NN	O	O
fusion	NN	O	O
with	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
nuclear	NN	O	O
contents	NN	O	O
of	NN	O	O
B	NN	O	B-protein
cell-expressed	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
are	NN	O	O
depleted	NN	O	O
within	NN	O	O
a	NN	O	O
few	NN	O	O
hours	NN	O	O
with	NN	O	O
kinetics	NN	O	O
that	NN	O	O
are	NN	O	O
as	NN	O	O
fast	NN	O	O
or	NN	O	O
faster	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
Ig	NN	O	B-DNA
gene	NN	O	I-DNA
extinction	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
extinguishing	NN	O	O
mechanism	NN	O	O
is	NN	O	O
effective	NN	O	O
very	NN	O	O
early	NN	O	O
after	NN	O	O
fusion	NN	O	O
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
extinction	NN	O	O
of	NN	O	O
Ig	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
global	NN	O	O
mechanism	NN	O	O
that	NN	O	O
suppresses	NN	O	O
the	NN	O	O
differentiation	NN	O	O
program	NN	O	O
foreign	NN	O	O
to	NN	O	O
the	NN	O	O
HeLa	NN	O	B-cell_line
phenotype	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Evidence	NN	O	O
for	NN	O	O
suppressed	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NFAT1	NN	O	B-protein
at	NN	O	O
its	NN	O	O
proximal	NN	O	B-DNA
binding	NN	O	I-DNA
element	NN	O	I-DNA
P0	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
enhanced	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
atopic	NN	O	O
patients	NN	O	O
.	NN	O	O

Allergen-specific	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
atopic	NN	O	O
patients	NN	O	O
are	NN	O	O
polarized	NN	O	O
IL-4-producing	NN	O	B-cell_line
Th2	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
promoting	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
by	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
increased	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
atopy	NN	O	O
is	NN	O	O
not	NN	O	O
fully	NN	O	O
understood	NN	O	O
.	NN	O	O

IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
regulation	NN	O	O
in	NN	O	O
general	NN	O	O
involves	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	I-protein
NFAT	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
of	NN	O	O
which	NN	O	O
NFAT1	NN	O	B-protein
and	NN	O	O
NFAT2	NN	O	B-protein
are	NN	O	O
most	NN	O	O
prominent	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
a	NN	O	O
unique	NN	O	O
inhibitory	NN	O	O
role	NN	O	O
of	NN	O	O
NFAT1	NN	O	B-protein
in	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
control	NN	O	O
was	NN	O	O
shown	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
with	NN	O	O
protein	NN	O	O
extracts	NN	O	O
of	NN	O	O
highly	NN	O	B-cell_line
polarized	NN	O	I-cell_line
Th2	NN	O	I-cell_line
clones	NN	O	I-cell_line
from	NN	O	O
atopics	NN	O	O
and	NN	O	O
Th1	NN	O	B-cell_line
clones	NN	O	I-cell_line
from	NN	O	O
controls	NN	O	O
we	NN	O	O
compared	NN	O	O
DNA-binding	NN	O	O
activities	NN	O	O
at	NN	O	O
the	NN	O	O
two	NN	O	O
NFAT-binding	NN	O	B-DNA
elements	NN	O	I-DNA
P0	NN	O	B-DNA
and	NN	O	O
P1	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
crucial	NN	O	O
proximal	NN	O	O
human	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

At	NN	O	O
the	NN	O	O
most	NN	O	O
proximal	NN	O	O
P0	NN	O	B-DNA
site	NN	O	O
,	NN	O	O
NFAT-containing	NN	O	O
complexes	NN	O	O
devoid	NN	O	O
of	NN	O	O
NFAT2	NN	O	B-protein
were	NN	O	O
readily	NN	O	O
inducible	NN	O	O
in	NN	O	O
the	NN	O	O
Th1	NN	O	B-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
hardly	NN	O	O
or	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
Th2	NN	O	B-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
both	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_line
and	NN	O	I-cell_line
Th2	NN	O	I-cell_line
clones	NN	O	I-cell_line
NFAT-containing	NN	O	O
complexes	NN	O	O
were	NN	O	O
strongly	NN	O	O
inducible	NN	O	O
at	NN	O	O
the	NN	O	O
P1	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
consisting	NN	O	O
of	NN	O	O
NFAT2	NN	O	B-protein
and	NN	O	O
a	NN	O	O
P0	NN	O	B-DNA
-compatible	NN	O	O
NFAT	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
without	NN	O	O
apparent	NN	O	O
differences	NN	O	O
between	NN	O	O
Th1	NN	O	B-cell_line
and	NN	O	I-cell_line
Th2	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

Like	NN	O	O
in	NN	O	O
Th2	NN	O	B-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
suppressed	NN	O	O
NFAT-	NN	O	O
P0	NN	O	B-DNA
complex	NN	O	O
formation	NN	O	O
was	NN	O	O
observed	NN	O	O
also	NN	O	O
at	NN	O	O
the	NN	O	O
polyclonal	NN	O	O
level	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
of	NN	O	O
three	NN	O	O
of	NN	O	O
five	NN	O	O
severe	NN	O	O
atopic	NN	O	O
dermatitis	NN	O	O
patients	NN	O	O
with	NN	O	O
strongly	NN	O	O
elevated	NN	O	O
serum	NN	O	O
IgE	NN	O	B-protein
levels	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
control	NN	O	O
PBMC	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
high-level	NN	O	O
IL-4	NN	O	B-protein
production	NN	O	O
in	NN	O	O
atopic	NN	O	B-cell_type
Th2	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
selective	NN	O	O
reduction	NN	O	O
of	NN	O	O
suppressive	NN	O	O
NFAT1	NN	O	B-protein
activity	NN	O	O
at	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
P0	NN	O	I-DNA
element	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
some	NN	O	O
patients	NN	O	O
with	NN	O	O
this	NN	O	O
multifactorial	NN	O	O
disease	NN	O	O
may	NN	O	O
have	NN	O	O
a	NN	O	O
putative	NN	O	O
systemic	NN	O	O
disorder	NN	O	O
at	NN	O	O
this	NN	O	O
level	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
expression	NN	O	O
and	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CTCF	NN	O	B-protein
,	NN	O	O
a	NN	O	O
c-myc	NN	O	O
transcriptional	NN	O	O
regulator	NN	O	O
,	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	O
myeloid	NN	O	O
cells	NN	O	O
.	NN	O	O

CTCF	NN	O	B-protein
is	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
repressor	NN	O	O
of	NN	O	O
the	NN	O	O
c-myc	NN	O	O
gene	NN	O	O
.	NN	O	O

Although	NN	O	O
CTCF	NN	O	B-protein
has	NN	O	O
been	NN	O	O
characterized	NN	O	O
in	NN	O	O
some	NN	O	O
detail	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
very	NN	O	O
little	NN	O	O
information	NN	O	O
about	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
CTCF	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Therefore	NN	O	O
we	NN	O	O
investigated	NN	O	O
CTCF	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
phosphorylation	NN	O	O
during	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
:	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
both	NN	O	O
CTCF	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
are	NN	O	O
down-regulated	NN	O	O
during	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
in	NN	O	O
most	NN	O	O
cell	NN	O	O
lines	NN	O	O
tested	NN	O	O
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
CTCF	NN	O	B-protein
down-regulation	NN	O	O
is	NN	O	O
retarded	NN	O	O
and	NN	O	O
less	NN	O	O
pronounced	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
c-myc	NN	O	B-DNA
;	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
CTCF	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
differentially	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
profiles	NN	O	O
depend	NN	O	O
on	NN	O	O
the	NN	O	O
differentiation	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
concluded	NN	O	O
that	NN	O	O
CTCF	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
activity	NN	O	O
is	NN	O	O
controlled	NN	O	O
at	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
post-transcriptional	NN	O	O
levels	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	O
resistance	NN	O	O
in	NN	O	O
the	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
immunophilin	NN	O	B-protein
FKBP51	NN	O	I-protein
.	NN	O	O

Squirrel	NN	O	O
monkeys	NN	O	O
are	NN	O	O
neotropical	NN	O	O
primates	NN	O	O
that	NN	O	O
have	NN	O	O
high	NN	O	O
circulating	NN	O	O
cortisol	NN	O	O
to	NN	O	O
compensate	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GRs	NN	O	B-protein
)	NN	O	O
with	NN	O	O
reduced	NN	O	O
affinity	NN	O	O
.	NN	O	O

The	NN	O	O
low	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
GR	NN	O	B-protein
does	NN	O	O
not	NN	O	O
result	NN	O	O
from	NN	O	O
substitutions	NN	O	O
in	NN	O	O
the	NN	O	O
receptor	NN	O	O
,	NN	O	O
because	NN	O	O
squirrel	NN	O	B-protein
monkey	NN	O	I-protein
GR	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
vitro	NN	O	O
exhibits	NN	O	O
high	NN	O	O
affinity	NN	O	O
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
squirrel	NN	O	O
monkeys	NN	O	O
express	NN	O	O
a	NN	O	O
soluble	NN	O	O
factor	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
mixing	NN	O	O
studies	NN	O	O
of	NN	O	O
cytosol	NN	O	O
from	NN	O	O
squirrel	NN	O	B-cell_type
monkey	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
SML	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
mouse	NN	O	O
L929	NN	O	O
cells	NN	O	O
,	NN	O	O
reduced	NN	O	O
GR	NN	O	O
binding	NN	O	O
affinity	NN	O	O
by	NN	O	O
11-fold	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
effort	NN	O	O
to	NN	O	O
identify	NN	O	O
this	NN	O	O
factor	NN	O	O
,	NN	O	O
the	NN	O	O
cellular	NN	O	O
levels	NN	O	O
of	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
heterocomplex	NN	O	I-protein
in	NN	O	O
SML	NN	O	B-cell_type
and	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
HL	NN	O	B-cell_type
)	NN	O	O
were	NN	O	O
compared	NN	O	O
.	NN	O	O

The	NN	O	O
immunophilin	NN	O	B-protein
FKBP51	NN	O	I-protein
was	NN	O	O
13-fold	NN	O	O
higher	NN	O	O
in	NN	O	O
SML	NN	O	B-cell_type
than	NN	O	O
in	NN	O	O
HL	NN	O	O
cytosol	NN	O	O
;	NN	O	O
FKBP52	NN	O	B-protein
in	NN	O	O
SML	NN	O	B-cell_type
was	NN	O	O
42	NN	O	O
%	NN	O	O
of	NN	O	O
that	NN	O	O
in	NN	O	O
HL	NN	O	O
cytosol	NN	O	O
.	NN	O	O

A	NN	O	O
role	NN	O	O
for	NN	O	O
changes	NN	O	O
in	NN	O	O
immunophilins	NN	O	B-protein
,	NN	O	O
causing	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
in	NN	O	O
neotropical	NN	O	O
primates	NN	O	O
,	NN	O	O
is	NN	O	O
supported	NN	O	O
by	NN	O	O
the	NN	O	O
following	NN	O	O
:	NN	O	O
the	NN	O	O
changes	NN	O	O
in	NN	O	O
FKBP51	NN	O	B-protein
and	NN	O	O
FKBP52	NN	O	B-protein
were	NN	O	O
observed	NN	O	O
in	NN	O	O
cells	NN	O	O
from	NN	O	O
other	NN	O	O
neotropical	NN	O	O
primates	NN	O	O
with	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
;	NN	O	O
the	NN	O	O
elevated	NN	O	O
level	NN	O	O
of	NN	O	O
FKBP51	NN	O	B-protein
was	NN	O	O
reflected	NN	O	O
in	NN	O	O
an	NN	O	O
abundance	NN	O	O
of	NN	O	O
FKBP51	NN	O	B-protein
in	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
protein	NN	O	I-protein
90	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
SML	NN	O	B-cell_type
;	NN	O	O
when	NN	O	O
cytosols	NN	O	O
of	NN	O	O
SML	NN	O	B-cell_type
and	NN	O	O
L929	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
mixed	NN	O	O
,	NN	O	O
the	NN	O	O
decrease	NN	O	O
in	NN	O	O
GR	NN	O	O
binding	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
incorporation	NN	O	O
of	NN	O	O
FKBP51	NN	O	B-protein
into	NN	O	O
GR	NN	O	B-protein
heterocomplexes	NN	O	I-protein
;	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
SML	NN	O	B-cell_type
cytosol	NN	O	O
on	NN	O	O
GR	NN	O	O
binding	NN	O	O
was	NN	O	O
reproduced	NN	O	O
with	NN	O	O
cytosol	NN	O	O
from	NN	O	O
COS	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
FKBP51	NN	O	B-protein
;	NN	O	O
and	NN	O	O
both	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
SML	NN	O	B-cell_type
cytosol	NN	O	O
on	NN	O	O
GR	NN	O	O
binding	NN	O	O
and	NN	O	O
the	NN	O	O
incorporation	NN	O	O
of	NN	O	O
FKBP51	NN	O	B-protein
into	NN	O	O
GR	NN	O	B-protein
heterocomplexes	NN	O	I-protein
were	NN	O	O
blocked	NN	O	O
by	NN	O	O
FK506	NN	O	O
.	NN	O	O

Regulation	NN	O	O
of	NN	O	O
GR	NN	O	O
binding	NN	O	O
by	NN	O	O
FKBP51	NN	O	B-protein
represents	NN	O	O
a	NN	O	O
previously	NN	O	O
unrecognized	NN	O	O
mechanism	NN	O	O
for	NN	O	O
regulating	NN	O	O
glucocorticoid	NN	O	O
sensitivity	NN	O	O
.	NN	O	O

-DOCSTART-	O

High-level	NN	O	O
replication	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
requires	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
through	NN	O	O
interaction	NN	O	O
with	NN	O	O
thymic	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
interaction	NN	O	O
of	NN	O	O
infected	NN	O	O
thymocytes	NN	O	B-cell_type
with	NN	O	O
autologous	NN	O	O
thymic	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
TEC	NN	O	B-cell_type
)	NN	O	O
is	NN	O	O
a	NN	O	O
prerequisite	NN	O	O
for	NN	O	O
a	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
replication	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
(	NN	O	O
M.Rothe	NN	O	O
,	NN	O	O
L.Chene	NN	O	O
,	NN	O	O
M.Nugeyre	NN	O	O
,	NN	O	O
F.Barre-Sinoussi	NN	O	O
,	NN	O	O
and	NN	O	O
N.Israel	NN	O	O
,	NN	O	O
J.Virol.72	NN	O	O
:	NN	O	O
5852-5861	NN	O	O
,	NN	O	O
1998	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
this	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV	NN	O	O
replication	NN	O	O
takes	NN	O	O
place	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
Rel/NF-kappaB	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
first	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
an	NN	O	O
HIV-1	NN	O	O
provirus	NN	O	O
(	NN	O	O
SF-2	NN	O	O
strain	NN	O	O
)	NN	O	O
very	NN	O	O
effectively	NN	O	O
replicates	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
cocultured	NN	O	O
with	NN	O	O
TEC	NN	O	B-cell_type
whereas	NN	O	O
this	NN	O	O
provirus	NN	O	O
,	NN	O	O
with	NN	O	O
kappaB	NN	O	B-DNA
sites	NN	O	I-DNA
deleted	NN	O	O
,	NN	O	O
fails	NN	O	O
to	NN	O	O
replicate	NN	O	O
.	NN	O	O

We	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
several	NN	O	O
NF-kappaB	NN	O	B-protein
complexes	NN	O	I-protein
are	NN	O	O
constitutively	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
either	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
thymus	NN	O	O
or	NN	O	O
maintained	NN	O	O
in	NN	O	O
coculture	NN	O	O
with	NN	O	O
autologous	NN	O	O
or	NN	O	O
heterologous	NN	O	O
TEC	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
prevalent	NN	O	O
complex	NN	O	O
is	NN	O	O
the	NN	O	O
heterodimer	NN	O	B-protein
p50-p65	NN	O	I-protein
.	NN	O	O

NF-kappaB	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
tightly	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
transfected	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
cotransfection	NN	O	O
of	NN	O	O
this	NN	O	O
LTR	NN	O	B-DNA
with	NN	O	O
a	NN	O	O
mutated	NN	O	O
IkappaBalpha	NN	O	B-protein
molecule	NN	O	I-protein
formally	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
LTR	NN	O	B-DNA
transactivation	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Rel/NF-kappaB	NN	O	B-protein
family	NN	O	I-protein
in	NN	O	O
thymocytes	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
also	NN	O	O
showed	NN	O	O
that	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
secreted	NN	O	O
within	NN	O	O
the	NN	O	O
coculture	NN	O	O
,	NN	O	O
induce	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
and	NN	O	O
a	NN	O	O
correlative	NN	O	O
LTR	NN	O	B-DNA
transactivation	NN	O	O
.	NN	O	O

However	NN	O	O
IL-7	NN	O	B-protein
,	NN	O	O
a	NN	O	O
crucial	NN	O	O
factor	NN	O	O
for	NN	O	O
thymopoiesis	NN	O	O
that	NN	O	O
is	NN	O	O
secreted	NN	O	O
mainly	NN	O	O
by	NN	O	O
TEC	NN	O	B-cell_type
,	NN	O	O
is	NN	O	O
a	NN	O	O
necessary	NN	O	O
cofactor	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
elicited	NN	O	O
by	NN	O	O
TNF	NN	O	B-protein
or	NN	O	O
IL-1	NN	O	B-protein
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
required	NN	O	O
for	NN	O	O
a	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
,	NN	O	O
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
a	NN	O	O
specific	NN	O	O
manner	NN	O	O
in	NN	O	O
the	NN	O	O
thymic	NN	O	O
microenvironment	NN	O	O
which	NN	O	O
provides	NN	O	O
the	NN	O	O
necessary	NN	O	O
cytokines	NN	O	O
:	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
IL-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-7	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Imbalanced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
isoforms	NN	O	I-protein
in	NN	O	O
cultured	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
systemic	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
and	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
is	NN	O	O
expressed	NN	O	O
as	NN	O	O
two	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
isoforms	NN	O	O
,	NN	O	O
GRalpha	NN	O	B-protein
and	NN	O	O
GRbeta	NN	O	B-protein
.	NN	O	O

Whereas	NN	O	O
GRalpha	NN	O	B-protein
is	NN	O	O
a	NN	O	O
hormone-activated	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
GRbeta	NN	O	B-protein
does	NN	O	O
not	NN	O	O
bind	NN	O	O
glucocorticoids	NN	O	O
(	NN	O	O
GCs	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
transcriptionally	NN	O	O
inactive	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
a	NN	O	O
potential	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
activated	NN	O	O
GRalpha	NN	O	B-protein
.	NN	O	O

Differential	NN	O	O
expression	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
isoforms	NN	O	I-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
generalized	NN	O	O
or	NN	O	O
tissue-specific	NN	O	O
GC	NN	O	O
resistance	NN	O	O
.	NN	O	O

GCs	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
neoplastic	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
and	NN	O	O
,	NN	O	O
defective	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
genesis	NN	O	O
of	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CLL	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
generalized	NN	O	O
GC	NN	O	O
resistance	NN	O	O
and	NN	O	O
CLL	NN	O	O
.	NN	O	O

GR	NN	O	O
number	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
transformed	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
was	NN	O	O
approximately	NN	O	O
one	NN	O	O
half	NN	O	O
that	NN	O	O
of	NN	O	O
control	NN	O	O
cells	NN	O	O
with	NN	O	O
a	NN	O	O
approximately	NN	O	O
10-fold	NN	O	O
reduction	NN	O	O
in	NN	O	O
binding	NN	O	O
affinity	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
apoptosis	NN	O	O
induction	NN	O	O
in	NN	O	O
CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
delayed	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
GCs	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
to	NN	O	O
other	NN	O	O
apoptosis	NN	O	O
inducers	NN	O	O
.	NN	O	O

Sequencing	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-DNA
cDNA	NN	O	I-DNA
and	NN	O	O
gene	NN	O	O
including	NN	O	O
the	NN	O	O
2.3-kb	NN	O	O
coding	NN	O	O
region	NN	O	O
,	NN	O	O
the	NN	O	O
intron/exon	NN	O	O
junctions	NN	O	O
,	NN	O	O
the	NN	O	O
known	NN	O	O
5'-regulatory	NN	O	B-DNA
region	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
approximately	NN	O	O
300	NN	O	O
bp	NN	O	O
of	NN	O	O
the	NN	O	O
3'-region	NN	O	B-DNA
revealed	NN	O	O
no	NN	O	O
alterations	NN	O	O
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
with	NN	O	O
an	NN	O	O
N-terminal	NN	O	O
antibody	NN	O	O
showed	NN	O	O
normal	NN	O	O
levels	NN	O	O
of	NN	O	O
immunoreactive	NN	O	B-protein
GR	NN	O	I-protein
,	NN	O	O
but	NN	O	O
quantitative	NN	O	O
analysis	NN	O	O
with	NN	O	O
isoform-specific	NN	O	O
C-terminal	NN	O	B-protein
antibodies	NN	O	I-protein
revealed	NN	O	O
a	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
GRalpha	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
high	NN	O	O
GRbeta	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
imbalanced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
isoforms	NN	O	I-protein
may	NN	O	O
be	NN	O	O
a	NN	O	O
mechanism	NN	O	O
of	NN	O	O
GC	NN	O	O
resistance	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
tumorigenesis	NN	O	O
by	NN	O	O
enhancing	NN	O	O
cell	NN	O	O
survival	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1999	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Intranuclear	NN	O	O
targeted	NN	O	O
delivery	NN	O	O
of	NN	O	O
functional	NN	O	O
NF-kappaB	NN	O	B-protein
by	NN	O	O
70	NN	O	B-protein
kDa	NN	O	I-protein
heat	NN	O	I-protein
shock	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
70	NN	O	B-protein
kDa	NN	O	I-protein
heat	NN	O	I-protein
shock	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
Hsp70	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
,	NN	O	O
ubiquitous	NN	O	B-protein
protein	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
chaperoning	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
various	NN	O	O
cellular	NN	O	O
organelles	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
when	NN	O	O
added	NN	O	O
exogenously	NN	O	O
to	NN	O	O
cells	NN	O	O
,	NN	O	O
Hsp70	NN	O	B-protein
is	NN	O	O
readily	NN	O	O
imported	NN	O	O
into	NN	O	O
both	NN	O	O
cytoplasmic	NN	O	O
and	NN	O	O
nuclear	NN	O	O
compartments	NN	O	O
in	NN	O	O
a	NN	O	O
cell-type-specific	NN	O	O
fashion	NN	O	O
.	NN	O	O

We	NN	O	O
exploited	NN	O	O
this	NN	O	O
ability	NN	O	O
of	NN	O	O
Hsp70	NN	O	B-protein
to	NN	O	O
deliver	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
a	NN	O	O
key	NN	O	O
transcriptional	NN	O	O
regulator	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
fusion	NN	O	O
protein	NN	O	O
composed	NN	O	O
of	NN	O	O
a	NN	O	O
C-terminal	NN	O	O
Hsp70	NN	O	B-protein
peptide	NN	O	O
and	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappaB	NN	O	B-protein
was	NN	O	O
directed	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
cells	NN	O	O
,	NN	O	O
could	NN	O	O
bind	NN	O	O
DNA	NN	O	O
specifically	NN	O	O
,	NN	O	O
and	NN	O	O
activated	NN	O	O
Igkappa	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
TNFalpha	NN	O	B-protein
production	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
propose	NN	O	O
that	NN	O	O
Hsp70	NN	O	B-protein
can	NN	O	O
be	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
vehicle	NN	O	O
for	NN	O	O
intracytoplasmic	NN	O	O
and	NN	O	O
intranuclear	NN	O	O
delivery	NN	O	O
of	NN	O	O
proteins	NN	O	O
or	NN	O	O
DNA	NN	O	O
to	NN	O	O
modulate	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
thereby	NN	O	O
control	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
elk-1	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Potential	NN	O	O
role	NN	O	O
of	NN	O	O
a	NN	O	O
downstream	NN	O	B-DNA
intronic	NN	O	I-DNA
sequence	NN	O	I-DNA
for	NN	O	O
elk-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

To	NN	O	O
characterize	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
elk-1	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
mapped	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
isolated	NN	O	O
elk-1-specific	NN	O	B-cell_line
genomic	NN	O	I-cell_line
phage	NN	O	I-cell_line
clones	NN	O	I-cell_line
that	NN	O	O
contained	NN	O	O
extensive	NN	O	O
upstream	NN	O	B-DNA
and	NN	O	I-DNA
downstream	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
TATA-like	NN	O	B-DNA
motif	NN	O	I-DNA
was	NN	O	O
identified	NN	O	O
immediately	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Functional	NN	O	O
analyses	NN	O	O
of	NN	O	O
DNA	NN	O	B-DNA
fragments	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
element	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
DNase	NN	O	B-DNA
I-hypersensitive	NN	O	I-DNA
chromatin	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
HS	NN	O	B-DNA
1	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
close	NN	O	O
proximity	NN	O	O
to	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
identified	NN	O	O
TATA	NN	O	B-DNA
motif	NN	O	I-DNA
is	NN	O	O
important	NN	O	O
for	NN	O	O
elk-1	NN	O	B-DNA
transcription	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Sequences	NN	O	O
upstream	NN	O	O
and	NN	O	O
downstream	NN	O	O
from	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
were	NN	O	O
found	NN	O	O
to	NN	O	O
contribute	NN	O	O
to	NN	O	O
elk-1	NN	O	B-DNA
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
hypersensitive	NN	O	O
site	NN	O	O
(	NN	O	O
HS	NN	O	O
2	NN	O	O
)	NN	O	O
was	NN	O	O
identified	NN	O	O
within	NN	O	O
the	NN	O	O
first	NN	O	O
intron	NN	O	O
in	NN	O	O
pre-monocytic	NN	O	O
cells	NN	O	O
,	NN	O	O
which	NN	O	O
express	NN	O	O
Elk-1	NN	O	O
only	NN	O	O
when	NN	O	O
differentiating	NN	O	O
to	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
which	NN	O	O
display	NN	O	O
a	NN	O	O
constitutive	NN	O	O
Elk-1	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
HS	NN	O	O
2	NN	O	O
did	NN	O	O
not	NN	O	O
exist	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
inducibility	NN	O	O
of	NN	O	O
elk-1	NN	O	B-DNA
expression	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
HS	NN	O	O
2	NN	O	O
.	NN	O	O

Egr-1	NN	O	O
and	NN	O	O
the	NN	O	O
serum	NN	O	O
response	NN	O	O
factor	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
interact	NN	O	O
specifically	NN	O	O
with	NN	O	O
the	NN	O	O
intronic	NN	O	B-DNA
sequence	NN	O	I-DNA
at	NN	O	O
+265	NN	O	O
and	NN	O	O
+448	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Because	NN	O	O
Egr-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
levels	NN	O	O
were	NN	O	O
observed	NN	O	O
to	NN	O	O
increase	NN	O	O
significantly	NN	O	O
before	NN	O	O
induction	NN	O	O
of	NN	O	O
elk-1	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
Egr-1	NN	O	B-protein
is	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
elk-1	NN	O	B-DNA
transcription	NN	O	O
in	NN	O	O
differentiating	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Immunohistochemical	NN	O	O
study	NN	O	O
of	NN	O	O
c-fos-positive	NN	O	B-protein
lymphocytes	NN	O	I-protein
infiltrated	NN	O	O
into	NN	O	O
human	NN	O	O
squamous	NN	O	O
cell	NN	O	O
carcinomas	NN	O	O
of	NN	O	O
the	NN	O	O
head	NN	O	O
and	NN	O	O
neck	NN	O	O
during	NN	O	O
radiation	NN	O	O
therapy	NN	O	O
and	NN	O	O
its	NN	O	O
clinical	NN	O	O
significance	NN	O	O
.	NN	O	O

C-fos	NN	O	B-DNA
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
systems	NN	O	O
after	NN	O	O
many	NN	O	O
kinds	NN	O	O
of	NN	O	O
stresses	NN	O	O
,	NN	O	O
including	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
.	NN	O	O

Changes	NN	O	O
in	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
radiation	NN	O	O
therapy	NN	O	O
in	NN	O	O
tumor	NN	O	O
tissues	NN	O	O
have	NN	O	O
not	NN	O	O
yet	NN	O	O
been	NN	O	O
reported	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
attempted	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
radiotherapy	NN	O	O
in	NN	O	O
human	NN	O	O
squamous	NN	O	O
cell	NN	O	O
carcinomas	NN	O	O
of	NN	O	O
the	NN	O	O
head	NN	O	O
and	NN	O	O
neck	NN	O	O
and	NN	O	O
to	NN	O	O
establish	NN	O	O
a	NN	O	O
possible	NN	O	O
correlation	NN	O	O
between	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
and	NN	O	O
the	NN	O	O
therapeutic	NN	O	O
effects	NN	O	O
of	NN	O	O
radiation	NN	O	O
therapy	NN	O	O
.	NN	O	O

Twenty-seven	NN	O	O
patients	NN	O	O
with	NN	O	O
tumors	NN	O	O
of	NN	O	O
the	NN	O	O
oral	NN	O	O
cavity	NN	O	O
,	NN	O	O
oropharynx	NN	O	O
,	NN	O	O
and	NN	O	O
maxillary	NN	O	O
sinus	NN	O	O
were	NN	O	O
examined	NN	O	O
,	NN	O	O
all	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
confirmed	NN	O	O
as	NN	O	O
squamous	NN	O	O
cell	NN	O	O
carcinomas	NN	O	O
.	NN	O	O

After	NN	O	O
obtaining	NN	O	O
the	NN	O	O
patients	NN	O	O
'	NN	O	O
informed	NN	O	O
consent	NN	O	O
,	NN	O	O
biopsies	NN	O	O
were	NN	O	O
performed	NN	O	O
before	NN	O	O
treatment	NN	O	O
and	NN	O	O
at	NN	O	O
doses	NN	O	O
of	NN	O	O
4	NN	O	O
,	NN	O	O
10	NN	O	O
,	NN	O	O
and	NN	O	O
20	NN	O	O
Gy	NN	O	O
of	NN	O	O
radiotherapy	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
specimens	NN	O	O
were	NN	O	O
preserved	NN	O	O
in	NN	O	O
liquid	NN	O	O
nitrogen	NN	O	O
for	NN	O	O
further	NN	O	O
examination	NN	O	O
.	NN	O	O

Serial	NN	O	O
sectioning	NN	O	O
of	NN	O	O
6	NN	O	O
micrometer	NN	O	O
was	NN	O	O
performed	NN	O	O
using	NN	O	O
a	NN	O	O
cryostat	NN	O	O
,	NN	O	O
and	NN	O	O
samples	NN	O	O
were	NN	O	O
immunohistochemically	NN	O	O
stained	NN	O	O
using	NN	O	O
the	NN	O	O
streptoavidin	NN	O	B-protein
-biotin	NN	O	B-protein
peroxidase	NN	O	I-protein
method	NN	O	O
and	NN	O	O
a	NN	O	O
monoclonal	NN	O	O
antibody	NN	O	O
against	NN	O	O
c-fos	NN	O	O
.	NN	O	O

Three	NN	O	O
of	NN	O	O
the	NN	O	O
27	NN	O	O
patients	NN	O	O
with	NN	O	O
squamous	NN	O	O
cell	NN	O	O
carcinoma	NN	O	O
showed	NN	O	O
slight	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	O
in	NN	O	O
their	NN	O	O
tumor	NN	O	O
cells	NN	O	O
before	NN	O	O
and/or	NN	O	O
at	NN	O	O
4	NN	O	O
or	NN	O	O
10	NN	O	O
Gy	NN	O	O
of	NN	O	O
radiotherapy	NN	O	O
.	NN	O	O

The	NN	O	O
tumors	NN	O	O
showed	NN	O	O
high	NN	O	O
radiosensitivity	NN	O	O
.	NN	O	O

Concerning	NN	O	O
tumor-infiltrating	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
moderate	NN	O	O
or	NN	O	O
remarkable	NN	O	O
grades	NN	O	O
of	NN	O	O
c-fos-positive	NN	O	B-protein
lymphocytes	NN	O	I-protein
before	NN	O	O
radiotherapy	NN	O	O
and	NN	O	O
at	NN	O	O
radiation	NN	O	O
doses	NN	O	O
of	NN	O	O
4	NN	O	O
,	NN	O	O
10	NN	O	O
,	NN	O	O
and	NN	O	O
20	NN	O	O
Gy	NN	O	O
was	NN	O	O
8.0	NN	O	O
,	NN	O	O
29.2	NN	O	O
,	NN	O	O
4.8	NN	O	O
,	NN	O	O
and	NN	O	O
0	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
immunohistochemical	NN	O	O
findings	NN	O	O
and	NN	O	O
the	NN	O	O
antitumor	NN	O	O
effect	NN	O	O
at	NN	O	O
a	NN	O	O
radiation	NN	O	O
dose	NN	O	O
of	NN	O	O
20	NN	O	O
Gy	NN	O	O
was	NN	O	O
examined	NN	O	O
on	NN	O	O
the	NN	O	O
corresponding	NN	O	O
H	NN	O	O
&	NN	O	O
E-stained	NN	O	O
sections	NN	O	O
.	NN	O	O

In	NN	O	O
patients	NN	O	O
whose	NN	O	O
infiltration	NN	O	O
of	NN	O	O
c-fos-positive	NN	O	B-protein
lymphocytes	NN	O	I-protein
into	NN	O	O
tumor	NN	O	O
tissues	NN	O	O
were	NN	O	O
moderate	NN	O	O
or	NN	O	O
remarkable	NN	O	O
at	NN	O	O
4	NN	O	O
Gy	NN	O	O
of	NN	O	O
radiotherapy	NN	O	O
,	NN	O	O
the	NN	O	O
tumors	NN	O	O
responded	NN	O	O
significantly	NN	O	O
well	NN	O	O
to	NN	O	O
radiation	NN	O	O
therapy	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.025	NN	O	O
,	NN	O	O
chi2	NN	O	O
test	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
patients	NN	O	O
took	NN	O	O
a	NN	O	O
significantly	NN	O	O
favorable	NN	O	O
clinical	NN	O	O
course	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
,	NN	O	O
chi2	NN	O	O
test	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
sample	NN	O	O
from	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
,	NN	O	O
c-fos-positive	NN	O	B-protein
lymphocytes	NN	O	I-protein
were	NN	O	O
identified	NN	O	O
as	NN	O	O
CD4	NN	O	O
positive	NN	O	O
and	NN	O	O
CD8	NN	O	O
negative	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
high	NN	O	O
radiosensitivity	NN	O	O
of	NN	O	O
squamous	NN	O	O
cell	NN	O	O
carcinomas	NN	O	O
in	NN	O	O
our	NN	O	O
samples	NN	O	O
could	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
an	NN	O	O
overexpression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-protein
in	NN	O	O
the	NN	O	O
tumor-infiltrating	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
induced	NN	O	O
by	NN	O	O
small	NN	O	O
doses	NN	O	O
of	NN	O	O
radiation	NN	O	O
therapy	NN	O	O
,	NN	O	O
and	NN	O	O
these	NN	O	O
activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
exerted	NN	O	O
a	NN	O	O
cytotoxic	NN	O	O
effect	NN	O	O
against	NN	O	O
the	NN	O	O
cancer	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Paternal	NN	O	O
expression	NN	O	O
of	NN	O	O
WT1	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
and	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
Wilms	NN	O	B-DNA
'	NN	O	I-DNA
tumor	NN	O	I-DNA
suppressor	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
WT1	NN	O	B-protein
)	NN	O	O
was	NN	O	O
previously	NN	O	O
identified	NN	O	O
as	NN	O	O
being	NN	O	O
imprinted	NN	O	O
,	NN	O	O
with	NN	O	O
frequent	NN	O	O
maternal	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	O
placentae	NN	O	O
and	NN	O	O
fetal	NN	O	O
brains	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
allele-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
WT1	NN	O	B-protein
in	NN	O	O
cultured	NN	O	O
human	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
from	NN	O	O
15	NN	O	O
individuals	NN	O	O
.	NN	O	O

Seven	NN	O	O
of	NN	O	O
15	NN	O	O
fibroblast	NN	O	B-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
polymorphic	NN	O	B-DNA
alleles	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
patterns	NN	O	O
were	NN	O	O
variable	NN	O	O
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
equal	NN	O	O
,	NN	O	O
unequal	NN	O	O
or	NN	O	O
monoallelic	NN	O	O
paternal	NN	O	O
expression	NN	O	O
in	NN	O	O
three	NN	O	O
,	NN	O	O
two	NN	O	O
and	NN	O	O
two	NN	O	O
cases	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Exclusive	NN	O	O
paternal	NN	O	O
expression	NN	O	O
of	NN	O	O
WT1	NN	O	B-protein
was	NN	O	O
also	NN	O	O
shown	NN	O	O
in	NN	O	O
non-cultured	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
latter	NN	O	O
two	NN	O	O
individuals	NN	O	O
.	NN	O	O

The	NN	O	O
allele-specific	NN	O	O
expression	NN	O	O
profiles	NN	O	O
of	NN	O	O
other	NN	O	O
imprinted	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
IGF2	NN	O	B-DNA
and	NN	O	O
H19	NN	O	B-DNA
,	NN	O	O
on	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
11	NN	O	I-DNA
were	NN	O	O
constant	NN	O	O
and	NN	O	O
consistent	NN	O	O
with	NN	O	O
those	NN	O	O
in	NN	O	O
other	NN	O	O
tissues	NN	O	O
.	NN	O	O

Our	NN	O	O
unexpected	NN	O	O
observations	NN	O	O
of	NN	O	O
paternal	NN	O	O
or	NN	O	O
biallelic	NN	O	O
expression	NN	O	O
of	NN	O	O
WT1	NN	O	B-protein
in	NN	O	O
fibroblasts	NN	O	B-cell_type
and	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
together	NN	O	O
with	NN	O	O
the	NN	O	O
previous	NN	O	O
findings	NN	O	O
of	NN	O	O
maternal	NN	O	O
or	NN	O	O
biallelic	NN	O	O
expression	NN	O	O
in	NN	O	O
placentae	NN	O	O
and	NN	O	O
brains	NN	O	O
,	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
allele-specific	NN	O	O
regulatory	NN	O	O
system	NN	O	O
of	NN	O	O
WT1	NN	O	B-protein
is	NN	O	O
unique	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
putative	NN	O	O
tissue-	NN	O	O
and	NN	O	O
individual-specific	NN	O	O
modifier	NN	O	O
.	NN	O	O

-DOCSTART-	O

T	NN	O	O
cell	NN	O	O
reactivity	NN	O	O
to	NN	O	O
Sjogren	NN	O	O
's	NN	O	O
syndrome	NN	O	O
related	NN	O	O
antigen	NN	O	O
La	NN	O	B-protein
(	NN	O	I-protein
SSB	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

OBJECTIVE	NN	O	O
.	NN	O	O

Many	NN	O	O
patients	NN	O	O
with	NN	O	O
primary	NN	O	O
Sjogren	NN	O	O
's	NN	O	O
syndrome	NN	O	O
(	NN	O	O
SS	NN	O	O
)	NN	O	O
make	NN	O	O
high	NN	O	O
titer	NN	O	O
IgG	NN	O	B-protein
autoantibodies	NN	O	I-protein
to	NN	O	O
the	NN	O	O
La	NN	O	B-protein
(	NN	O	I-protein
SSB	NN	O	I-protein
)	NN	O	I-protein
antigen	NN	O	O
,	NN	O	O
suggesting	NN	O	O
antigen	NN	O	O
specific	NN	O	O
T	NN	O	O
cell-B	NN	O	O
cell	NN	O	O
interactions	NN	O	O
.	NN	O	O

T	NN	O	O
cell	NN	O	O
responses	NN	O	O
to	NN	O	O
some	NN	O	O
nuclear	NN	O	O
antigens	NN	O	O
,	NN	O	O
particularly	NN	O	O
U1RNP	NN	O	B-protein
,	NN	O	O
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
(	NN	O	O
SLE	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
T	NN	O	O
cell	NN	O	O
reactivity	NN	O	O
to	NN	O	O
the	NN	O	O
autoantigen	NN	O	B-protein
SSB	NN	O	I-protein
in	NN	O	O
patients	NN	O	O
with	NN	O	O
SS	NN	O	O
and	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

METHODS	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
proliferation	NN	O	O
assay	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
reactivity	NN	O	O
to	NN	O	O
purified	NN	O	B-protein
recombinant	NN	O	I-protein
SSB	NN	O	I-protein
(	NN	O	O
rSSB	NN	O	B-protein
)	NN	O	O
in	NN	O	O
20	NN	O	O
patients	NN	O	O
with	NN	O	O
SS	NN	O	O
and	NN	O	O
19	NN	O	O
controls	NN	O	O
.	NN	O	O

Specificity	NN	O	O
was	NN	O	O
determined	NN	O	O
using	NN	O	O
tetanus	NN	O	O
toxoid	NN	O	O
,	NN	O	O
endotoxin	NN	O	O
,	NN	O	O
and	NN	O	O
3	NN	O	O
other	NN	O	O
autoantigens	NN	O	B-protein
(	NN	O	O
PBC.M2	NN	O	B-protein
,	NN	O	O
Sc170	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GAD	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Precursor	NN	O	O
frequency	NN	O	O
was	NN	O	O
calculated	NN	O	O
by	NN	O	O
limiting	NN	O	O
dilution	NN	O	O
analysis	NN	O	O
.	NN	O	O

HLA	NN	O	O
Class	NN	O	O
II	NN	O	O
dependency	NN	O	O
was	NN	O	O
investigated	NN	O	O
using	NN	O	O
anti-Class	NN	O	B-protein
II	NN	O	I-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

HLA-DR	NN	O	O
typing	NN	O	O
was	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
and	NN	O	O
sequence	NN	O	O
specific	NN	O	O
oligonucleotide	NN	O	O
typing	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
.	NN	O	O

Six	NN	O	O
of	NN	O	O
20	NN	O	O
patients	NN	O	O
with	NN	O	O
SS	NN	O	O
and	NN	O	O
10/19	NN	O	O
controls	NN	O	O
proliferated	NN	O	O
to	NN	O	O
La	NN	O	O
(	NN	O	O
rSSB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Precursor	NN	O	O
frequency	NN	O	O
of	NN	O	O
anti-SSB	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
1	NN	O	O
:	NN	O	O
77	NN	O	O
,	NN	O	O
040	NN	O	O
and	NN	O	O
1	NN	O	O
:	NN	O	O
115	NN	O	O
,	NN	O	O
000	NN	O	O
in	NN	O	O
2	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
and	NN	O	O
1	NN	O	O
:	NN	O	O
230	NN	O	O
,	NN	O	O
250	NN	O	O
and	NN	O	O
1	NN	O	O
:	NN	O	O
103	NN	O	O
,	NN	O	O
034	NN	O	O
in	NN	O	O
two	NN	O	O
patients	NN	O	O
with	NN	O	O
SS	NN	O	O
.	NN	O	O

Anti-	NN	O	O
HLA-DR	NN	O	B-protein
abrogated	NN	O	O
proliferation	NN	O	O
to	NN	O	O
SSB	NN	O	B-protein
and	NN	O	O
tetanus	NN	O	O
toxoid	NN	O	O
.	NN	O	O

Thirteen	NN	O	O
of	NN	O	O
15	NN	O	O
patients	NN	O	O
with	NN	O	O
SS	NN	O	O
and	NN	O	O
4/17	NN	O	O
controls	NN	O	O
were	NN	O	O
HLA-DR3	NN	O	B-protein
positive	NN	O	O
,	NN	O	O
with	NN	O	O
no	NN	O	O
apparent	NN	O	O
association	NN	O	O
of	NN	O	O
HLA-DR3	NN	O	B-protein
with	NN	O	O
SSB	NN	O	B-protein
reactivity	NN	O	O
in	NN	O	O
controls	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
.	NN	O	O

Anti-La	NN	O	B-cell_line
(	NN	O	I-cell_line
SSB	NN	O	I-cell_line
)	NN	O	I-cell_line
specific	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
occur	NN	O	O
in	NN	O	O
a	NN	O	O
significant	NN	O	O
proportion	NN	O	O
of	NN	O	O
controls	NN	O	O
and	NN	O	O
in	NN	O	O
some	NN	O	O
patients	NN	O	O
with	NN	O	O
SS	NN	O	O
.	NN	O	O

The	NN	O	O
function	NN	O	O
of	NN	O	O
SSB	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
controls	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
defined	NN	O	O
.	NN	O	O

They	NN	O	O
may	NN	O	O
represent	NN	O	O
immunoregulatory	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
further	NN	O	O
analysis	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
comparison	NN	O	O
to	NN	O	O
those	NN	O	O
found	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
SS	NN	O	O
,	NN	O	O
may	NN	O	O
elucidate	NN	O	O
normal	NN	O	O
immunoregulation	NN	O	O
and	NN	O	O
the	NN	O	O
derangements	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
Sjogren	NN	O	O
's	NN	O	O
syndrome	NN	O	O
.	NN	O	O

-DOCSTART-	O

Anti-rheumatic	NN	O	O
compound	NN	O	O
aurothioglucose	NN	O	O
inhibits	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
-induced	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
latently	NN	O	O
infected	NN	O	O
OM10.1	NN	O	B-cell_line
and	NN	O	I-cell_line
Ach2	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

NF-kappaB	NN	O	B-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
cellular	NN	O	O
activator	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Down-regulation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
inhibit	NN	O	O
HIV	NN	O	O
replication	NN	O	O
from	NN	O	O
the	NN	O	O
latently	NN	O	B-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Gold	NN	O	O
compounds	NN	O	O
have	NN	O	O
been	NN	O	O
effectively	NN	O	O
used	NN	O	O
for	NN	O	O
many	NN	O	O
decades	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
gold	NN	O	O
compounds	NN	O	O
,	NN	O	O
especially	NN	O	O
aurothioglucose	NN	O	O
(	NN	O	O
AuTG	NN	O	O
)	NN	O	O
containing	NN	O	O
monovalent	NN	O	O
gold	NN	O	O
ion	NN	O	O
,	NN	O	O
inhibited	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
efficacy	NN	O	O
of	NN	O	O
the	NN	O	O
gold	NN	O	O
compound	NN	O	O
AuTG	NN	O	O
as	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
latently	NN	O	O
infected	NN	O	O
OM10.1	NN	O	B-cell_line
and	NN	O	I-cell_line
Ach2	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
-alpha	NN	O	I-protein
-induced	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
OM10.1	NN	O	B-cell_line
or	NN	O	O
Ach2	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
non-cytotoxic	NN	O	O
doses	NN	O	O
of	NN	O	O
AuTG	NN	O	O
(	NN	O	O
>	NN	O	O
10	NN	O	O
microM	NN	O	O
in	NN	O	O
OM10.1	NN	O	O
cells	NN	O	O
and	NN	O	O
>	NN	O	O
25	NN	O	O
F.M	NN	O	O
in	NN	O	O
Ach2	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
while	NN	O	O
25	NN	O	O
microM	NN	O	O
of	NN	O	O
the	NN	O	O
counter-anion	NN	O	B-protein
thioglucose	NN	O	I-protein
(	NN	O	O
TG	NN	O	B-protein
)	NN	O	O
or	NN	O	O
gold	NN	O	O
compound	NN	O	O
containing	NN	O	O
divalent	NN	O	O
gold	NN	O	O
ion	NN	O	O
,	NN	O	O
HAuCl3	NN	O	O
,	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
AuTG	NN	O	O
on	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
a	NN	O	O
transient	NN	O	O
CAT	NN	O	B-protein
assay	NN	O	O
.	NN	O	O

Specific	NN	O	O
staining	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
electron	NN	O	O
microscopic	NN	O	O
examinations	NN	O	O
revealed	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
metal	NN	O	O
gold	NN	O	O
in	NN	O	O
the	NN	O	O
cells	NN	O	O
,	NN	O	O
supporting	NN	O	O
our	NN	O	O
previous	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
gold	NN	O	O
ions	NN	O	O
could	NN	O	O
block	NN	O	O
NF-kappaB	NN	O	B-protein
-DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
a	NN	O	O
redox	NN	O	O
mechanism	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
monovalent	NN	O	O
gold	NN	O	O
compound	NN	O	O
AuTG	NN	O	O
is	NN	O	O
a	NN	O	O
potentially	NN	O	O
useful	NN	O	O
drug	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
patients	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV	NN	O	O
.	NN	O	O

-DOCSTART-	O

Clonality	NN	O	O
of	NN	O	O
isolated	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
hypereosinophilic	NN	O	O
syndrome	NN	O	O
.	NN	O	O

The	NN	O	O
idiopathic	NN	O	O
hypereosinophilic	NN	O	O
syndrome	NN	O	O
(	NN	O	O
IHES	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
rare	NN	O	O
disorder	NN	O	O
characterized	NN	O	O
by	NN	O	O
unexplained	NN	O	O
,	NN	O	O
persistent	NN	O	O
eosinophilia	NN	O	O
associated	NN	O	O
with	NN	O	O
multiple	NN	O	O
organ	NN	O	O
dysfunction	NN	O	O
due	NN	O	O
to	NN	O	O
eosinophilic	NN	O	O
tissue	NN	O	O
infiltration	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
karyotypic	NN	O	O
abnormalities	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
specific	NN	O	O
test	NN	O	O
to	NN	O	O
detect	NN	O	O
clonal	NN	O	O
eosinophilia	NN	O	O
in	NN	O	O
IHES	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
X-chromosome	NN	O	B-DNA
inactivation	NN	O	O
patterns	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
proliferative	NN	O	O
disorders	NN	O	O
are	NN	O	O
clonal	NN	O	O
in	NN	O	O
origin	NN	O	O
.	NN	O	O

Methylation	NN	O	O
of	NN	O	O
HpaII	NN	O	B-DNA
and	NN	O	I-DNA
Hha	NN	O	I-DNA
I	NN	O	I-DNA
sites	NN	O	I-DNA
near	NN	O	O
the	NN	O	O
polymorphic	NN	O	B-DNA
trinucleotide	NN	O	I-DNA
repeat	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
androgen	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
HUMARA	NN	O	B-DNA
)	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
correlate	NN	O	O
with	NN	O	O
X-inactivation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
used	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
with	NN	O	O
nested	NN	O	O
primers	NN	O	O
to	NN	O	O
analyze	NN	O	O
X-inactivation	NN	O	O
patterns	NN	O	O
of	NN	O	O
the	NN	O	O
HUMARA	NN	O	B-DNA
loci	NN	O	I-DNA
in	NN	O	O
purified	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
from	NN	O	O
female	NN	O	O
patients	NN	O	O
with	NN	O	O
eosinophilia	NN	O	O
.	NN	O	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
eosinophils	NN	O	I-cell_type
were	NN	O	O
isolated	NN	O	O
by	NN	O	O
their	NN	O	O
autofluoresence	NN	O	O
using	NN	O	O
flow	NN	O	O
cytometric	NN	O	O
sorting	NN	O	O
.	NN	O	O

Eosinophils	NN	O	B-cell_type
purified	NN	O	O
from	NN	O	O
a	NN	O	O
female	NN	O	O
patient	NN	O	O
presenting	NN	O	O
with	NN	O	O
IHES	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
show	NN	O	O
a	NN	O	O
clonal	NN	O	O
pattern	NN	O	O
of	NN	O	O
X-inactivation	NN	O	O
.	NN	O	O

Eosinophil-depleted	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
from	NN	O	O
this	NN	O	O
patient	NN	O	O
were	NN	O	O
polyclonal	NN	O	O
by	NN	O	O
HUMARA	NN	O	B-DNA
analysis	NN	O	O
,	NN	O	O
thus	NN	O	O
excluding	NN	O	O
skewedness	NN	O	O
of	NN	O	O
random	NN	O	O
X-inactivation	NN	O	O
.	NN	O	O

After	NN	O	O
corticosteroid	NN	O	O
suppression	NN	O	O
of	NN	O	O
her	NN	O	O
blood	NN	O	O
eosinophilia	NN	O	O
,	NN	O	O
a	NN	O	O
clonal	NN	O	O
population	NN	O	O
of	NN	O	O
eosinophils	NN	O	B-cell_type
could	NN	O	O
no	NN	O	O
longer	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
purified	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
eosinophils	NN	O	B-cell_type
purified	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
Churg-Strauss	NN	O	O
syndrome	NN	O	O
and	NN	O	O
from	NN	O	O
six	NN	O	O
patients	NN	O	O
with	NN	O	O
reactive	NN	O	O
eosinophilias	NN	O	O
attributed	NN	O	O
to	NN	O	O
allergy	NN	O	O
,	NN	O	O
parasitic	NN	O	O
infection	NN	O	O
,	NN	O	O
or	NN	O	O
drug	NN	O	O
reaction	NN	O	O
showed	NN	O	O
a	NN	O	O
polyclonal	NN	O	O
pattern	NN	O	O
of	NN	O	O
X-inactivation	NN	O	O
by	NN	O	O
HUMARA	NN	O	B-DNA
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
finding	NN	O	O
of	NN	O	O
clonal	NN	O	O
eosinophilia	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
presenting	NN	O	O
with	NN	O	O
IHES	NN	O	O
indicates	NN	O	O
that	NN	O	O
such	NN	O	O
patients	NN	O	O
may	NN	O	O
have	NN	O	O
,	NN	O	O
in	NN	O	O
reality	NN	O	O
,	NN	O	O
a	NN	O	O
low-grade	NN	O	O
clonal	NN	O	O
disorder	NN	O	O
that	NN	O	O
can	NN	O	O
be	NN	O	O
distinguished	NN	O	O
from	NN	O	O
reactive	NN	O	O
eosinophilias	NN	O	O
by	NN	O	O
HUMARA	NN	O	B-DNA
analysis	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
the	NN	O	O
method	NN	O	O
described	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
monitor	NN	O	O
disease	NN	O	O
progression	NN	O	O
.	NN	O	O

-DOCSTART-	O

GA-binding	NN	O	B-protein
protein	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
the	NN	O	O
coactivator	NN	O	B-protein
CREB	NN	O	I-protein
binding	NN	O	I-protein
protein/p300	NN	O	I-protein
,	NN	O	O
control	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
16	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Interleukin	NN	O	B-protein
16	NN	O	I-protein
(	NN	O	O
IL-16	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
chemotactic	NN	O	B-protein
cytokine	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
affects	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
replication	NN	O	O
of	NN	O	O
HIV	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
expressed	NN	O	O
as	NN	O	O
a	NN	O	O
large	NN	O	O
67-kDa	NN	O	B-protein
precursor	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
pro-IL-16	NN	O	B-protein
)	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
airway	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
asthmatics	NN	O	O
after	NN	O	O
challenge	NN	O	O
with	NN	O	O
allergen	NN	O	O
.	NN	O	O

This	NN	O	O
pro-IL-16	NN	O	B-protein
is	NN	O	O
subsequently	NN	O	O
processed	NN	O	O
to	NN	O	O
the	NN	O	O
mature	NN	O	O
cytokine	NN	O	O
of	NN	O	O
13	NN	O	B-protein
kDa	NN	O	I-protein
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-16	NN	O	B-protein
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
,	NN	O	O
we	NN	O	O
cloned	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
chromosomal	NN	O	I-DNA
IL-16	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
analyzed	NN	O	O
its	NN	O	O
promoter	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
IL-16	NN	O	I-DNA
gene	NN	O	I-DNA
consists	NN	O	O
of	NN	O	O
seven	NN	O	O
exons	NN	O	B-DNA
and	NN	O	O
six	NN	O	O
introns	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
sequences	NN	O	I-DNA
up	NN	O	O
to	NN	O	O
nucleotide	NN	O	O
-120	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
and	NN	O	O
murine	NN	O	O
IL-16	NN	O	B-protein
genes	NN	O	O
share	NN	O	O
>	NN	O	O
84	NN	O	O
%	NN	O	O
sequence	NN	O	O
homology	NN	O	O
and	NN	O	O
harbor	NN	O	B-DNA
promoter	NN	O	I-DNA
elements	NN	O	I-DNA
for	NN	O	O
constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
both	NN	O	O
promoters	NN	O	O
lack	NN	O	O
any	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
they	NN	O	O
contain	NN	O	O
two	NN	O	O
CAAT	NN	O	B-DNA
box-like	NN	O	I-DNA
motifs	NN	O	I-DNA
and	NN	O	O
three	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
of	NN	O	O
GA-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
GABP	NN	O	I-protein
)	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Two	NN	O	O
of	NN	O	O
these	NN	O	O
motifs	NN	O	O
are	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
highly	NN	O	O
conserved	NN	O	O
and	NN	O	O
inducible	NN	O	O
dyad	NN	O	B-DNA
symmetry	NN	O	I-DNA
element	NN	O	I-DNA
shown	NN	O	O
previously	NN	O	O
to	NN	O	O
control	NN	O	O
a	NN	O	O
remote	NN	O	O
IL-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
CD18	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
concert	NN	O	O
with	NN	O	O
the	NN	O	O
coactivator	NN	O	B-protein
CREB	NN	O	I-protein
binding	NN	O	I-protein
protein/p300	NN	O	I-protein
,	NN	O	O
which	NN	O	O
interacts	NN	O	O
with	NN	O	O
GABPalpha	NN	O	B-protein
,	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
GABPalpha	NN	O	B-protein
and	NN	O	O
-beta	NN	O	B-protein
to	NN	O	O
the	NN	O	O
dyad	NN	O	B-DNA
symmetry	NN	O	I-DNA
element	NN	O	I-DNA
controls	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-16	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Supplementing	NN	O	O
the	NN	O	O
data	NN	O	O
on	NN	O	O
the	NN	O	O
processing	NN	O	O
of	NN	O	O
pro-IL-16	NN	O	B-protein
,	NN	O	O
our	NN	O	O
results	NN	O	O
indicate	NN	O	O
the	NN	O	O
complexity	NN	O	O
of	NN	O	O
IL-16	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
tightly	NN	O	O
controlled	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
posttranslational	NN	O	O
levels	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Tuberculosis	NN	O	O
and	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
virus	NN	O	O
infection	NN	O	O
in	NN	O	O
Africans	NN	O	O
and	NN	O	O
variation	NN	O	O
in	NN	O	O
the	NN	O	O
vitamin	NN	O	B-DNA
D	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
active	NN	O	O
metabolite	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
dihydroxyvitamin	NN	O	O
D3	NN	O	O
,	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
immunoregulatory	NN	O	O
hormone	NN	O	O
[	NN	O	O
1	NN	O	O
]	NN	O	O
.	NN	O	O

Its	NN	O	O
effects	NN	O	O
are	NN	O	O
exerted	NN	O	O
by	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
present	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Variation	NN	O	O
in	NN	O	O
the	NN	O	O
vitamin	NN	O	B-DNA
D	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
typed	NN	O	O
in	NN	O	O
2015	NN	O	O
subjects	NN	O	O
from	NN	O	O
large	NN	O	O
case-control	NN	O	O
studies	NN	O	O
of	NN	O	O
three	NN	O	O
major	NN	O	O
infectious	NN	O	O
diseases	NN	O	O
:	NN	O	O
tuberculosis	NN	O	O
,	NN	O	O
malaria	NN	O	O
,	NN	O	O
and	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
virus	NN	O	O
.	NN	O	O

Homozygotes	NN	O	O
for	NN	O	O
a	NN	O	O
polymorphism	NN	O	O
at	NN	O	O
codon	NN	O	B-DNA
352	NN	O	I-DNA
(	NN	O	O
genotype	NN	O	O
tt	NN	O	O
)	NN	O	O
were	NN	O	O
significantly	NN	O	O
underrepresented	NN	O	O
among	NN	O	O
those	NN	O	O
with	NN	O	O
tuberculosis	NN	O	O
(	NN	O	O
chi2=6.22	NN	O	O
,	NN	O	O
1	NN	O	O
df	NN	O	O
,	NN	O	O
P=.01	NN	O	O
)	NN	O	O
and	NN	O	O
persistent	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
infection	NN	O	O
(	NN	O	O
chi2=6.25	NN	O	O
,	NN	O	O
1	NN	O	O
df	NN	O	O
,	NN	O	O
P=.01	NN	O	O
)	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
subjects	NN	O	O
with	NN	O	O
clinical	NN	O	O
malaria	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
other	NN	O	O
genotypes	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
this	NN	O	O
genetic	NN	O	O
variant	NN	O	O
,	NN	O	O
which	NN	O	O
predisposes	NN	O	O
to	NN	O	O
low	NN	O	O
bone	NN	O	O
mineral	NN	O	O
density	NN	O	O
in	NN	O	O
many	NN	O	O
populations	NN	O	O
,	NN	O	O
may	NN	O	O
confer	NN	O	O
resistance	NN	O	O
to	NN	O	O
certain	NN	O	O
infectious	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
:	NN	O	O
role	NN	O	O
of	NN	O	O
26S	NN	O	B-protein
proteasome	NN	O	I-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
becoming	NN	O	O
increasingly	NN	O	O
apparent	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
.	NN	O	O

Data	NN	O	O
obtained	NN	O	O
from	NN	O	O
studies	NN	O	O
in	NN	O	O
our	NN	O	O
laboratories	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
proteasome	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
cascade	NN	O	O
by	NN	O	O
regulating	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
the	NN	O	O
availability	NN	O	O
of	NN	O	O
selective	NN	O	O
and	NN	O	O
orally	NN	O	O
active	NN	O	O
proteasome	NN	O	O
inhibitors	NN	O	O
should	NN	O	O
prove	NN	O	O
useful	NN	O	O
in	NN	O	O
delineating	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
proteasome	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
other	NN	O	O
pathophysiological	NN	O	O
conditions	NN	O	O
such	NN	O	O
as	NN	O	O
cancer	NN	O	O
and	NN	O	O
heart	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tissue	NN	O	O
factor	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
is	NN	O	O
suppressed	NN	O	O
by	NN	O	O
lysophosphatidylcholine	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
TF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
principal	NN	O	O
initiator	NN	O	O
of	NN	O	O
coagulation	NN	O	O
,	NN	O	O
is	NN	O	O
increased	NN	O	O
during	NN	O	O
inflammation	NN	O	O
and	NN	O	O
atherosclerosis	NN	O	O
.	NN	O	O

Both	NN	O	O
conditions	NN	O	O
are	NN	O	O
promoted	NN	O	O
by	NN	O	O
lysophosphatidylcholine	NN	O	O
(	NN	O	O
lysoPC	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
that	NN	O	O
lysoPC	NN	O	O
(	NN	O	O
1	NN	O	O
to	NN	O	O
10	NN	O	O
micromol/L	NN	O	O
)	NN	O	O
dose-dependently	NN	O	O
reduced	NN	O	O
TF	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
elicited	NN	O	O
by	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

Lysophosphatidylethanolamine	NN	O	O
(	NN	O	O
lysoPE	NN	O	O
)	NN	O	O
and	NN	O	O
other	NN	O	O
lysophospholipids	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
LPS-induced	NN	O	O
TF	NN	O	O
activity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

TF	NN	O	O
antigen	NN	O	O
expression	NN	O	O
as	NN	O	O
elicited	NN	O	O
by	NN	O	O
LPS	NN	O	O
was	NN	O	O
also	NN	O	O
lowered	NN	O	O
by	NN	O	O
lysoPC	NN	O	O
.	NN	O	O

Phospholipid	NN	O	O
analyses	NN	O	O
indicated	NN	O	O
a	NN	O	O
selective	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
lysoPC	NN	O	O
content	NN	O	O
of	NN	O	O
the	NN	O	O
monocytes	NN	O	B-cell_type
after	NN	O	O
preincubation	NN	O	O
with	NN	O	O
the	NN	O	O
lysophospholipid	NN	O	O
.	NN	O	O

LysoPC	NN	O	O
inhibited	NN	O	O
the	NN	O	O
TF	NN	O	O
activity	NN	O	O
of	NN	O	O
Mono	NN	O	B-cell_line
Mac-6	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
a	NN	O	O
similar	NN	O	O
extent	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

LPS	NN	O	O
binding	NN	O	O
to	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
receptors	NN	O	O
and	NN	O	O
internalization	NN	O	O
of	NN	O	O
LPS	NN	O	O
into	NN	O	O
monocytes	NN	O	B-cell_type
were	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
lysoPC	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
LPS-mediated	NN	O	O
nuclear	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB/Rel	NN	O	I-protein
to	NN	O	O
a	NN	O	O
TF-specific	NN	O	B-DNA
kappaB	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
lysoPC	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
TF	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
by	NN	O	O
LPS	NN	O	O
tended	NN	O	O
to	NN	O	O
be	NN	O	O
partially	NN	O	O
reduced	NN	O	O
by	NN	O	O
the	NN	O	O
lysophospholipid	NN	O	O
.	NN	O	O

Preincubation	NN	O	O
with	NN	O	O
lysoPC	NN	O	O
increased	NN	O	O
monocytic	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
adenylyl	NN	O	B-protein
cyclase	NN	O	I-protein
by	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
2'-deoxy-3'-adenosine	NN	O	B-protein
monophosphate	NN	O	I-protein
partially	NN	O	O
reversed	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
TF	NN	O	O
activity	NN	O	O
promoted	NN	O	O
by	NN	O	O
lysoPC	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
lysoPC	NN	O	O
markedly	NN	O	O
decreases	NN	O	O
LPS-mediated	NN	O	O
TF	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
effect	NN	O	O
probably	NN	O	O
being	NN	O	O
mediated	NN	O	O
by	NN	O	O
both	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
posttranscriptional	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

LysoPC	NN	O	O
may	NN	O	O
thus	NN	O	O
attenuate	NN	O	O
activation	NN	O	O
of	NN	O	O
coagulation	NN	O	O
during	NN	O	O
inflammation	NN	O	O
and	NN	O	O
atherosclerosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
fas-ligand	NN	O	O
expression	NN	O	O
during	NN	O	O
activation-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
via	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
.	NN	O	O

T	NN	O	O
cell	NN	O	O
receptor	NN	O	O
engagement	NN	O	O
activates	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
important	NN	O	O
for	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
regulation	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
this	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
also	NN	O	O
leads	NN	O	O
to	NN	O	O
activation-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
that	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
FasL	NN	O	O
transcription	NN	O	O
and	NN	O	O
expression	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
many	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
expressed	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
also	NN	O	O
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation-induced	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
a	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridoma	NN	O	I-cell_line
leads	NN	O	O
to	NN	O	O
decreased	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
upon	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
stimulation	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
FasL	NN	O	B-DNA
promoter	NN	O	I-DNA
that	NN	O	O
contributes	NN	O	O
to	NN	O	O
such	NN	O	O
regulation	NN	O	O
.	NN	O	O

Co-expression	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
(	NN	O	I-protein
Rel	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	I-protein
with	NN	O	O
the	NN	O	O
FasL	NN	O	B-DNA
promoter	NN	O	I-DNA
enhanced	NN	O	O
its	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
co-expression	NN	O	O
of	NN	O	O
IkappaB	NN	O	B-protein
dramatically	NN	O	O
inhibited	NN	O	O
the	NN	O	O
inducible	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
activation-induced	NN	O	O
FasL	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
define	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
mediating	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Promoter	NN	O	B-DNA
sequence	NN	O	I-DNA
,	NN	O	O
exon	NN	O	O
:	NN	O	O
intron	NN	O	O
structure	NN	O	O
,	NN	O	O
and	NN	O	O
synteny	NN	O	O
of	NN	O	O
genetic	NN	O	O
location	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
chicken	NN	O	B-protein
cytokine	NN	O	I-protein
with	NN	O	O
T-cell	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
is	NN	O	O
IL2	NN	O	B-protein
and	NN	O	O
not	NN	O	O
IL15	NN	O	B-protein
.	NN	O	O

The	NN	O	O
gene	NN	O	O
encoding	NN	O	O
a	NN	O	O
chicken	NN	O	B-protein
cytokine	NN	O	I-protein
with	NN	O	O
T-cell	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
was	NN	O	O
cloned	NN	O	O
,	NN	O	O
sequenced	NN	O	O
,	NN	O	O
and	NN	O	O
mapped	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
cytokine	NN	O	B-protein
is	NN	O	O
chicken	NN	O	O
IL2	NN	O	B-protein
and	NN	O	O
not	NN	O	O
IL15	NN	O	B-protein
.	NN	O	O

The	NN	O	O
exon	NN	O	O
:	NN	O	O
intron	NN	O	O
structure	NN	O	O
of	NN	O	O
chicken	NN	O	B-protein
IL2	NN	O	I-protein
corresponds	NN	O	O
almost	NN	O	O
exactly	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
mammalian	NN	O	O
IL2s	NN	O	B-protein
with	NN	O	O
the	NN	O	O
exceptions	NN	O	O
of	NN	O	O
exon	NN	O	B-DNA
2	NN	O	I-DNA
and	NN	O	O
introns	NN	O	B-DNA
2	NN	O	O
and	NN	O	O
3	NN	O	O
which	NN	O	O
are	NN	O	O
shorter	NN	O	O
.	NN	O	O

Chicken	NN	O	O
IL2	NN	O	B-protein
contains	NN	O	O
five	NN	O	O
repeats	NN	O	O
of	NN	O	O
the	NN	O	O
``	NN	O	O
instability	NN	O	O
''	NN	O	O
motif	NN	O	O
ATTTA	NN	O	O
in	NN	O	O
the	NN	O	O
3'untranslated	NN	O	B-DNA
region	NN	O	I-DNA
in	NN	O	O
exon	NN	O	B-DNA
4	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
is	NN	O	O
a	NN	O	O
single-copy	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
with	NN	O	O
neither	NN	O	O
structural	NN	O	O
(	NN	O	O
amino	NN	O	O
acid	NN	O	O
)	NN	O	O
nor	NN	O	O
promoter	NN	O	O
sequence	NN	O	O
polymorphisms	NN	O	O
identified	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
predicted	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
suggests	NN	O	O
that	NN	O	O
overall	NN	O	O
protein	NN	O	B-protein
structure	NN	O	I-protein
is	NN	O	O
conserved	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
receptor	NN	O	O
binding	NN	O	O
sites	NN	O	O
are	NN	O	O
not	NN	O	O
.	NN	O	O

A	NN	O	O
number	NN	O	O
of	NN	O	O
potential	NN	O	B-DNA
regulatory	NN	O	I-DNA
sequences	NN	O	I-DNA
similar	NN	O	O
to	NN	O	O
those	NN	O	O
found	NN	O	O
in	NN	O	O
mammals	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

These	NN	O	O
include	NN	O	O
(	NN	O	O
5'-3	NN	O	O
'	NN	O	O
)	NN	O	O
a	NN	O	O
composite	NN	O	O
NF-AT/	NN	O	B-DNA
``	NN	O	I-DNA
AP-1	NN	O	I-DNA
''	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
CD28	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
an	NN	O	O
AP-1	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
an	NN	O	O
NF-AT	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
part	NN	O	I-DNA
of	NN	O	I-DNA
an	NN	O	I-DNA
AP-1/octamer	NN	O	I-DNA
composite	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
mammalian	NN	O	B-protein
NF-kappaB	NN	O	I-protein
and	NN	O	O
octamer	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
seem	NN	O	O
to	NN	O	O
be	NN	O	O
absent	NN	O	O
,	NN	O	O
although	NN	O	O
there	NN	O	O
are	NN	O	O
alternative	NN	O	O
potential	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
octamer-binding	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
chicken	NN	O	B-DNA
IL2	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
close	NN	O	O
proximity	NN	O	O
to	NN	O	O
their	NN	O	O
mammalian	NN	O	B-DNA
homologues	NN	O	I-DNA
.	NN	O	O

Sequence	NN	O	O
comparisons	NN	O	O
also	NN	O	O
predict	NN	O	O
other	NN	O	O
potential	NN	O	O
transcription	NN	O	O
factor	NN	O	O
binding	NN	O	O
sites	NN	O	O
as	NN	O	O
yet	NN	O	O
undescribed	NN	O	O
in	NN	O	O
mammalian	NN	O	B-DNA
IL2	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
Taq	NN	O	B-DNA
I	NN	O	I-DNA
polymorphism	NN	O	I-DNA
was	NN	O	O
identified	NN	O	O
which	NN	O	O
enabled	NN	O	O
chicken	NN	O	O
IL2	NN	O	B-protein
to	NN	O	O
be	NN	O	O
mapped	NN	O	O
to	NN	O	O
chromosome	NN	O	B-DNA
4	NN	O	I-DNA
,	NN	O	O
linked	NN	O	O
to	NN	O	O
ANX5	NN	O	B-DNA
,	NN	O	O
with	NN	O	O
synteny	NN	O	O
with	NN	O	O
mouse	NN	O	B-DNA
chromosome	NN	O	I-DNA
3	NN	O	I-DNA
and	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
4	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
non-mammalian	NN	O	B-DNA
cytokine	NN	O	I-DNA
gene	NN	O	I-DNA
to	NN	O	O
be	NN	O	O
mapped	NN	O	O
.	NN	O	O

-DOCSTART-	O

Endothelial	NN	O	O
production	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
:	NN	O	O
modulation	NN	O	O
by	NN	O	O
heparin	NN	O	O
and	NN	O	O
consequences	NN	O	O
for	NN	O	O
mononuclear	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Heparin	NN	O	O
is	NN	O	O
a	NN	O	O
polyanionic	NN	O	O
glycosaminoglycan	NN	O	O
(	NN	O	O
GAG	NN	O	O
)	NN	O	O
that	NN	O	O
can	NN	O	O
bind	NN	O	O
with	NN	O	O
high	NN	O	O
affinity	NN	O	O
to	NN	O	O
a	NN	O	O
range	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
including	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
and	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
chemokine	NN	O	B-protein
superfamily	NN	O	I-protein
.	NN	O	O

This	NN	O	O
GAG	NN	O	O
also	NN	O	O
possesses	NN	O	O
immunomodulatory	NN	O	O
activity	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
can	NN	O	O
antagonize	NN	O	O
the	NN	O	O
capacity	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
MHC	NN	O	I-protein
antigen	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
up-regulate	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
,	NN	O	O
by	NN	O	O
cultured	NN	O	B-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
binding	NN	O	O
to	NN	O	O
cell-surface	NN	O	O
heparan	NN	O	O
sulphate	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
optimal	NN	O	O
activity	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
that	NN	O	O
free	NN	O	O
heparin	NN	O	O
competitively	NN	O	O
inhibits	NN	O	O
this	NN	O	O
sequestration	NN	O	O
process	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
performed	NN	O	O
to	NN	O	O
increase	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
activity	NN	O	O
of	NN	O	O
heparin	NN	O	O
by	NN	O	O
investigation	NN	O	O
of	NN	O	O
potential	NN	O	O
antagonism	NN	O	O
of	NN	O	O
the	NN	O	O
production	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
monocyte	NN	O	B-protein
chemotactic	NN	O	I-protein
peptide-1	NN	O	I-protein
(	NN	O	O
MCP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
chemokine	NN	O	B-protein
important	NN	O	O
for	NN	O	O
mononuclear	NN	O	O
leucocyte	NN	O	O
recruitment	NN	O	O
across	NN	O	O
vascular	NN	O	O
endothelium	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
mixture	NN	O	O
of	NN	O	O
heparin	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
inhibited	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	O
transducer	NN	O	O
and	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	O
protein	NN	O	O
,	NN	O	O
STAT-1	NN	O	B-protein
produced	NN	O	O
normally	NN	O	O
by	NN	O	O
treatment	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

An	NN	O	O
inhibition	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
production	NN	O	O
was	NN	O	O
observed	NN	O	O
that	NN	O	O
was	NN	O	O
specifically	NN	O	O
caused	NN	O	O
by	NN	O	O
mixture	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
with	NN	O	O
heparin-like	NN	O	O
,	NN	O	O
and	NN	O	O
therefore	NN	O	O
cytokine-binding	NN	O	O
,	NN	O	O
GAGs	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
also	NN	O	O
shown	NN	O	O
that	NN	O	O
mixture	NN	O	O
of	NN	O	O
heparin-like	NN	O	O
GAGs	NN	O	O
with	NN	O	O
MCP-1	NN	O	B-protein
inhibited	NN	O	O
the	NN	O	O
rapid	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
which	NN	O	O
is	NN	O	O
normally	NN	O	O
produced	NN	O	O
by	NN	O	O
treatment	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
leucocytes	NN	O	I-cell_type
with	NN	O	O
this	NN	O	O
chemokine	NN	O	B-protein
.	NN	O	O

Blockade	NN	O	O
of	NN	O	O
this	NN	O	O
intracellular	NN	O	O
signalling	NN	O	O
event	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
response	NN	O	O
towards	NN	O	O
MCP-1	NN	O	B-protein
.	NN	O	O

Results	NN	O	O
from	NN	O	O
this	NN	O	O
study	NN	O	O
indicate	NN	O	O
that	NN	O	O
soluble	NN	O	O
,	NN	O	O
heparin-like	NN	O	O
GAGs	NN	O	O
can	NN	O	O
block	NN	O	O
IFN-gamma	NN	O	B-protein
-dependent	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
production	NN	O	O
by	NN	O	O
cultured	NN	O	B-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
can	NN	O	O
also	NN	O	O
antagonize	NN	O	O
the	NN	O	O
leucocyte-activating	NN	O	O
and	NN	O	O
migration-promoting	NN	O	O
properties	NN	O	O
of	NN	O	O
pre-existing	NN	O	O
MCP-1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
activities	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
immunomodulatory	NN	O	O
properties	NN	O	O
of	NN	O	O
heparin	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Molecular-biologic	NN	O	O
aspects	NN	O	O
of	NN	O	O
interaction	NN	O	O
between	NN	O	O
nervous	NN	O	O
and	NN	O	O
immune	NN	O	O
systems	NN	O	O
]	NN	O	O

The	NN	O	O
problem	NN	O	O
of	NN	O	O
the	NN	O	O
neuro-immuno	NN	O	O
interactions	NN	O	O
on	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
trans-factors	NN	O	I-protein
,	NN	O	O
stimulating	NN	O	O
interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
discussed	NN	O	O
.	NN	O	O

The	NN	O	O
physico-chemical	NN	O	O
and	NN	O	O
functional	NN	O	O
parameters	NN	O	O
of	NN	O	O
the	NN	O	O
low	NN	O	B-protein
molecular	NN	O	I-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
SP	NN	O	B-protein
and	NN	O	O
BP-	NN	O	B-protein
14	NN	O	I-protein
,	NN	O	I-protein
18	NN	O	I-protein
,	NN	O	I-protein
19	NN	O	I-protein
kDs	NN	O	I-protein
)	NN	O	O
isolated	NN	O	O
from	NN	O	O
splenic	NN	O	B-cell_type
and	NN	O	I-cell_type
brain	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
immunized	NN	O	O
rats	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
to	NN	O	O
the	NN	O	O
regulatory	NN	O	O
region	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2mRNA	NN	O	B-RNA
synthesis	NN	O	O
in	NN	O	O
splenic	NN	O	B-cell_line
T-lymphocytes	NN	O	I-cell_line
culture	NN	O	I-cell_line
in	NN	O	O
normal	NN	O	O
conditions	NN	O	O
were	NN	O	O
shown	NN	O	O
.	NN	O	O

The	NN	O	O
protective	NN	O	O
effect	NN	O	O
of	NN	O	O
SP	NN	O	B-protein
and	NN	O	O
BP	NN	O	B-protein
on	NN	O	O
the	NN	O	O
IL-2mRNA	NN	O	B-RNA
synthesis	NN	O	O
in	NN	O	O
stressful	NN	O	O
conditions	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
T-cells	NN	O	B-cell_type
treatment	NN	O	O
with	NN	O	O
the	NN	O	O
CsA	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interferon-alpha	NN	O	B-protein
activates	NN	O	O
multiple	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
upregulates	NN	O	O
proliferation-associated	NN	O	O
IL-2Ralpha	NN	O	B-protein
,	NN	O	O
c-myc	NN	O	B-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
pim-1	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Interferon-alpha	NN	O	B-protein
(	NN	O	O
IFN-alpha	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
pleiotropic	NN	O	B-protein
cytokine	NN	O	I-protein
that	NN	O	O
has	NN	O	O
antiviral	NN	O	O
,	NN	O	O
antiproliferative	NN	O	O
,	NN	O	O
and	NN	O	O
immunoregulatory	NN	O	B-protein
functions	NN	O	O
.	NN	O	O

There	NN	O	O
is	NN	O	O
increasing	NN	O	O
evidence	NN	O	O
that	NN	O	O
IFN-alpha	NN	O	B-protein
has	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
T-cell	NN	O	O
biology	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2Ralpha	NN	O	B-protein
,	NN	O	O
c-myc	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
pim-1	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
anti-CD3-activated	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
-induced	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
IFN-alpha	NN	O	B-protein
,	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-15	NN	O	B-protein
upregulated	NN	O	O
IL-2Ralpha	NN	O	B-protein
,	NN	O	O
c-myc	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
pim-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

IFN-alpha	NN	O	B-protein
also	NN	O	O
sensitized	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
,	NN	O	O
further	NN	O	O
suggesting	NN	O	O
that	NN	O	O
IFN-alpha	NN	O	B-protein
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
T-cell	NN	O	O
mitogenesis	NN	O	O
.	NN	O	O

When	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
IL-2Ralpha	NN	O	B-protein
,	NN	O	O
pim-1	NN	O	B-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
IRF-1	NN	O	I-DNA
GAS	NN	O	I-DNA
elements	NN	O	I-DNA
after	NN	O	O
cytokine	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
IFN-alpha	NN	O	B-protein
-induced	NN	O	O
binding	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT4	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
STAT5	NN	O	B-protein
to	NN	O	O
all	NN	O	O
of	NN	O	O
these	NN	O	O
elements	NN	O	O
.	NN	O	O

Yet	NN	O	O
,	NN	O	O
IFN-alpha	NN	O	B-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
activate	NN	O	O
binding	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
to	NN	O	O
the	NN	O	O
high-affinity	NN	O	O
IFP53	NN	O	O
GAS	NN	O	O
site	NN	O	O
.	NN	O	O

IFN-alpha	NN	O	B-protein
enhanced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1	NN	O	O
,	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
STAT4	NN	O	B-protein
,	NN	O	O
STAT5a	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT5b	NN	O	B-protein
.	NN	O	O

IL-12	NN	O	B-protein
induced	NN	O	O
STAT4	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-15	NN	O	B-protein
induced	NN	O	O
STAT5	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
GAS	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
IFN-alpha	NN	O	B-protein
,	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-15	NN	O	B-protein
have	NN	O	O
overlapping	NN	O	O
activities	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
findings	NN	O	O
thus	NN	O	O
emphasize	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
IFN-alpha	NN	O	B-protein
as	NN	O	O
a	NN	O	O
T-cell	NN	O	B-protein
regulatory	NN	O	I-protein
cytokine	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Megakaryocyte/Platelet-specific	NN	O	B-DNA
enhancer	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
alpha2beta1	NN	O	B-DNA
integrin	NN	O	I-DNA
gene	NN	O	I-DNA
:	NN	O	O
two	NN	O	O
tandem	NN	O	O
AP1	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
signaling	NN	O	O
cascade	NN	O	O
.	NN	O	O

The	NN	O	O
alpha2beta1	NN	O	B-protein
integrin	NN	O	I-protein
,	NN	O	O
a	NN	O	O
collagen	NN	O	O
receptor	NN	O	O
on	NN	O	O
platelets	NN	O	B-cell_type
and	NN	O	O
megakaryocytes	NN	O	B-cell_type
,	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
normal	NN	O	O
platelet	NN	O	O
function	NN	O	O
.	NN	O	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
alpha2	NN	O	B-DNA
integrin	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
cells	NN	O	O
undergoing	NN	O	O
megakaryocytic	NN	O	O
differentiation	NN	O	O
requires	NN	O	O
a	NN	O	O
core	NN	O	B-DNA
promoter	NN	O	I-DNA
between	NN	O	O
bp	NN	O	O
-30	NN	O	O
and	NN	O	O
-92	NN	O	O
,	NN	O	O
a	NN	O	O
silencer	NN	O	B-DNA
between	NN	O	O
bp	NN	O	O
-92	NN	O	O
and	NN	O	O
-351	NN	O	O
,	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	B-DNA
enhancers	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
flank	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
identified	NN	O	O
a	NN	O	O
229-bp	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
flank	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
alpha2	NN	O	B-DNA
integrin	NN	O	I-DNA
gene	NN	O	I-DNA
required	NN	O	O
for	NN	O	O
high-level	NN	O	B-cell_type
enhancer	NN	O	I-cell_type
activity	NN	O	I-cell_type
in	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
megakaryocytic	NN	O	O
features	NN	O	O
.	NN	O	O

Two	NN	O	O
tandem	NN	O	O
AP1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
with	NN	O	O
dyad	NN	O	O
symmetry	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
and	NN	O	O
for	NN	O	O
DNA-protein	NN	O	B-protein
complex	NN	O	I-protein
formation	NN	O	O
with	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos/c-jun	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

The	NN	O	O
requirement	NN	O	O
for	NN	O	O
AP1	NN	O	B-protein
activation	NN	O	O
suggested	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
in	NN	O	O
regulating	NN	O	O
alpha2	NN	O	B-DNA
integrin	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
cascade	NN	O	O
with	NN	O	O
PD98059	NN	O	O
,	NN	O	O
a	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
kinase	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
prevented	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
alpha2	NN	O	B-protein
integrin	NN	O	I-protein
subunit	NN	O	I-protein
in	NN	O	O
cells	NN	O	O
induced	NN	O	O
to	NN	O	O
become	NN	O	O
megakaryocytic	NN	O	O
.	NN	O	O

We	NN	O	O
provide	NN	O	O
a	NN	O	O
model	NN	O	O
of	NN	O	O
megakaryocytic	NN	O	O
differentiation	NN	O	O
in	NN	O	O
which	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
alpha2	NN	O	B-DNA
integrin	NN	O	I-DNA
gene	NN	O	I-DNA
requires	NN	O	O
signaling	NN	O	O
via	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
pathway	NN	O	O
to	NN	O	O
activate	NN	O	O
two	NN	O	O
tandem	NN	O	O
AP1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
alpha2	NN	O	B-DNA
integrin	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
association	NN	O	O
of	NN	O	O
Nmi	NN	O	B-protein
with	NN	O	O
Stat5	NN	O	B-protein
and	NN	O	O
Stat1	NN	O	B-protein
in	NN	O	O
IL-2	NN	O	B-protein
-and	NN	O	O
IFNgamma	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
coiled-coil	NN	O	O
region	NN	O	O
of	NN	O	O
Stat5b	NN	O	B-protein
as	NN	O	O
the	NN	O	O
bait	NN	O	O
in	NN	O	O
a	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
screen	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
Nmi	NN	O	B-protein
,	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
unknown	NN	O	O
function	NN	O	O
previously	NN	O	O
reported	NN	O	O
as	NN	O	O
an	NN	O	O
N-Myc	NN	O	B-protein
interactor	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
show	NN	O	O
that	NN	O	O
Nmi	NN	O	B-protein
interacts	NN	O	O
with	NN	O	O
all	NN	O	O
STATs	NN	O	B-protein
except	NN	O	O
Stat2	NN	O	B-protein
.	NN	O	O

We	NN	O	O
evaluated	NN	O	O
two	NN	O	O
cytokine	NN	O	O
systems	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IFNgamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Nmi	NN	O	B-protein
augments	NN	O	O
STAT	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
Nmi	NN	O	B-protein
lacks	NN	O	O
an	NN	O	O
intrinsic	NN	O	O
transcriptional	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
;	NN	O	O
instead	NN	O	O
,	NN	O	O
Nmi	NN	O	B-protein
enhances	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
CBP/p300	NN	O	B-protein
coactivator	NN	O	I-protein
proteins	NN	O	I-protein
with	NN	O	O
Stat1	NN	O	B-protein
and	NN	O	O
Stat5	NN	O	B-protein
,	NN	O	O
and	NN	O	O
together	NN	O	O
with	NN	O	O
CBP/p300	NN	O	B-protein
can	NN	O	O
augment	NN	O	O
IL-2-	NN	O	O
and	NN	O	O
IFNgamma-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
not	NN	O	O
only	NN	O	O
reveal	NN	O	O
that	NN	O	O
Nmi	NN	O	B-protein
can	NN	O	O
potentiate	NN	O	O
STAT	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
suggest	NN	O	O
that	NN	O	O
it	NN	O	O
can	NN	O	O
augment	NN	O	O
coactivator	NN	O	O
protein	NN	O	O
recruitment	NN	O	O
to	NN	O	O
at	NN	O	O
least	NN	O	O
some	NN	O	O
members	NN	O	O
of	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
sequence-specific	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Lactobacilli	NN	O	O
and	NN	O	O
vaginal	NN	O	O
host	NN	O	O
defense	NN	O	O
:	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
,	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Lactobacilli	NN	O	O
,	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
vaginal	NN	O	O
flora	NN	O	O
,	NN	O	O
can	NN	O	O
activate	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
HIV	NN	O	I-DNA
)	NN	O	I-DNA
-1	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
and	NN	O	O
THP-1	NN	O	B-cell_line
macrophage	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
strongly	NN	O	O
enhanced	NN	O	O
by	NN	O	O
vanadate	NN	O	O
and	NN	O	O
inhibited	NN	O	O
by	NN	O	O
catalase	NN	O	B-protein
,	NN	O	O
implicating	NN	O	O
H2O2	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
activation	NN	O	O
in	NN	O	O
THP-1	NN	O	B-protein
cells	NN	O	I-protein
occurred	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
vanadate	NN	O	O
and	NN	O	O
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
catalase	NN	O	B-protein
.	NN	O	O

The	NN	O	O
active	NN	O	O
material	NN	O	O
partitioned	NN	O	O
into	NN	O	O
the	NN	O	O
phenol	NN	O	O
layer	NN	O	O
on	NN	O	O
hot	NN	O	O
aqueous	NN	O	O
phenol	NN	O	O
extraction	NN	O	O
.	NN	O	O

Lactobacilli	NN	O	O
also	NN	O	O
increased	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
-alphaand	NN	O	O
interleukin	NN	O	B-protein
-1betaproduction	NN	O	O
and	NN	O	O
activated	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
THP-1	NN	O	B-protein
cells	NN	O	I-protein
and	NN	O	O
increased	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
-alphaproduction	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	O
vaginal	NN	O	O
fluid	NN	O	O
specimens	NN	O	O
had	NN	O	O
comparable	NN	O	O
properties	NN	O	O
,	NN	O	O
which	NN	O	O
correlated	NN	O	O
with	NN	O	O
their	NN	O	O
bacterial	NN	O	O
content	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
the	NN	O	O
presence	NN	O	O
in	NN	O	O
vaginal	NN	O	O
fluid	NN	O	O
of	NN	O	O
agent	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
derived	NN	O	O
from	NN	O	O
indigenous	NN	O	O
bacteria	NN	O	O
that	NN	O	O
can	NN	O	O
activate	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
,	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
cells	NN	O	O
of	NN	O	O
macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
with	NN	O	O
possible	NN	O	O
influence	NN	O	O
on	NN	O	O
vaginal	NN	O	O
physiology	NN	O	O
and	NN	O	O
host	NN	O	O
defense	NN	O	O
.	NN	O	O

-DOCSTART-	O

T	NN	O	O
cell	NN	O	O
priming	NN	O	O
enhances	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
increasing	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
.	NN	O	O

The	NN	O	O
repetitive	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
priming	NN	O	O
)	NN	O	O
enhances	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
IL-4	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
others	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Molecular	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
selective	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
by	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
remain	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
priming	NN	O	O
of	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
selectively	NN	O	O
enhances	NN	O	O
IL-4	NN	O	B-protein
expression	NN	O	O
relative	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
mechanism	NN	O	O
involving	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	I-protein
NFAT	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

As	NN	O	O
detected	NN	O	O
by	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprinting	NN	O	O
,	NN	O	O
priming	NN	O	O
markedly	NN	O	O
increases	NN	O	O
the	NN	O	O
activation-dependent	NN	O	O
engagement	NN	O	O
of	NN	O	O
the	NN	O	O
P0	NN	O	B-DNA
and	NN	O	O
P1	NN	O	B-DNA
NFAT-binding	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
each	NN	O	O
proximal	NN	O	B-DNA
P	NN	O	I-DNA
element	NN	O	I-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
optimal	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

Activated	NN	O	O
primed	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
contain	NN	O	O
more	NN	O	O
NFAT1	NN	O	B-protein
and	NN	O	O
support	NN	O	O
greater	NN	O	O
NFAT	NN	O	B-protein
-directed	NN	O	O
transcription	NN	O	O
than	NN	O	O
unprimed	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
activator	NN	O	B-protein
protein	NN	O	I-protein
1	NN	O	I-protein
binding	NN	O	O
and	NN	O	O
activator	NN	O	B-protein
protein	NN	O	I-protein
1	NN	O	I-protein
-mediated	NN	O	O
transcription	NN	O	O
by	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
is	NN	O	O
similar	NN	O	O
.	NN	O	O

Increased	NN	O	O
expression	NN	O	O
of	NN	O	O
wild-type	NN	O	O
NFAT1	NN	O	B-protein
substantially	NN	O	O
increases	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
unprimed	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
NFAT1	NN	O	B-protein
may	NN	O	O
be	NN	O	O
limiting	NN	O	O
for	NN	O	O
IL-4	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
this	NN	O	O
cell	NN	O	O
type	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
a	NN	O	O
truncated	NN	O	O
form	NN	O	O
of	NN	O	O
NFAT1	NN	O	B-protein
acts	NN	O	O
as	NN	O	O
a	NN	O	O
dominant-negative	NN	O	O
,	NN	O	O
reducing	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
primed	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
confirming	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
NFAT	NN	O	I-protein
to	NN	O	O
increased	NN	O	O
IL-4	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
effector	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

NFAT1	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
major	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
member	NN	O	I-protein
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
initial	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
by	NN	O	O
primed	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
and	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
venous	NN	O	O
and	NN	O	O
arterial	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
.	NN	O	O

We	NN	O	O
compared	NN	O	O
U-937	NN	O	B-cell_line
cell	NN	O	I-cell_line
adhesion	NN	O	O
and	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
venous	NN	O	I-cell_type
(	NN	O	O
HUVECs	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
arterial	NN	O	B-cell_type
(	NN	O	I-cell_type
HUAECs	NN	O	I-cell_type
)	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
exposed	NN	O	O
to	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
interleukin-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

TNF	NN	O	B-protein
and	NN	O	O
LPS	NN	O	O
stimulated	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
(	NN	O	O
VCAM	NN	O	B-protein
)	NN	O	O
-1	NN	O	O
surface	NN	O	O
expression	NN	O	O
and	NN	O	O
adhesion	NN	O	O
of	NN	O	O
U-937	NN	O	B-cell_line
monocyte-like	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
HUVECs	NN	O	B-cell_type
but	NN	O	O
not	NN	O	O
to	NN	O	O
HUAECs	NN	O	B-cell_type
.	NN	O	O

Antibody	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
in	NN	O	O
HUVECs	NN	O	B-cell_type
at	NN	O	O
least	NN	O	O
75	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
adhesion	NN	O	O
response	NN	O	O
is	NN	O	O
VCAM-1	NN	O	B-protein
mediated	NN	O	O
.	NN	O	O

Interleukin-1	NN	O	B-protein
stimulated	NN	O	O
U-937	NN	O	B-cell_line
cell	NN	O	I-cell_line
adhesion	NN	O	O
to	NN	O	O
and	NN	O	O
VCAM-1	NN	O	B-protein
surface	NN	O	O
expression	NN	O	O
in	NN	O	O
both	NN	O	O
HUVECs	NN	O	B-cell_type
and	NN	O	O
HUAECs	NN	O	B-cell_type
.	NN	O	O

Pyrrolidinedithiocarbamate	NN	O	O
and	NN	O	O
the	NN	O	O
proteasome	NN	O	O
inhibitor	NN	O	O
MG-132	NN	O	O
blocked	NN	O	O
TNF	NN	O	B-protein
-and	NN	O	O
LPS-stimulated	NN	O	O
U-937	NN	O	B-cell_line
cell	NN	O	I-cell_line
adhesion	NN	O	O
to	NN	O	O
HUVECs	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
agents	NN	O	O
also	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
TNF	NN	O	B-protein
-and	NN	O	O
LPS-stimulated	NN	O	O
increases	NN	O	O
in	NN	O	O
HUVEC	NN	O	B-protein
surface	NN	O	I-protein
VCAM-1	NN	O	I-protein
.	NN	O	O

TNF	NN	O	B-protein
increased	NN	O	O
VCAM-1	NN	O	B-protein
protein	NN	O	O
and	NN	O	O
mRNA	NN	O	B-RNA
in	NN	O	O
HUVECs	NN	O	B-cell_type
that	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
pyrrolidinedithiocarbamate	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
neither	NN	O	O
TNF	NN	O	B-protein
or	NN	O	O
LPS	NN	O	O
stimulated	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
HUAECs	NN	O	B-cell_type
.	NN	O	O

TNF	NN	O	B-protein
stimulated	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
and	NN	O	O
E-selectin	NN	O	B-protein
in	NN	O	O
HUVECs	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
in	NN	O	O
HUAECs	NN	O	B-cell_type
,	NN	O	O
only	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
was	NN	O	O
increased	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
no	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
-stimulated	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
activation	NN	O	O
between	NN	O	O
HUVECs	NN	O	B-cell_type
and	NN	O	O
HUAECs	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
novel	NN	O	O
and	NN	O	O
striking	NN	O	O
insensitivity	NN	O	O
of	NN	O	O
arterial	NN	O	O
endothelium	NN	O	O
to	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
and	NN	O	O
LPS	NN	O	O
and	NN	O	O
indicate	NN	O	O
a	NN	O	O
dissociation	NN	O	O
between	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
HUAECs	NN	O	B-cell_type
to	NN	O	O
upregulate	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
and	NN	O	O
VCAM-1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
testosterone	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
plasma	NN	O	O
membranes	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
testosterone	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
androgen	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
AR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
testosterone	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
murine	NN	O	B-cell_type
splenic	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
unconventional	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
for	NN	O	O
testosterone	NN	O	O
and	NN	O	O
classical	NN	O	B-protein
intracellular	NN	O	I-protein
AR	NN	O	I-protein
.	NN	O	O

Binding	NN	O	O
sites	NN	O	O
for	NN	O	O
testosterone	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
both	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	I-cell_type
CD8	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
subsets	NN	O	I-cell_type
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
directly	NN	O	O
revealed	NN	O	O
with	NN	O	O
the	NN	O	O
impeded	NN	O	O
ligand	NN	O	O
testosterone-BSA-FITC	NN	O	O
by	NN	O	O
confocal	NN	O	O
laser	NN	O	O
scanning	NN	O	O
microscopy	NN	O	O
(	NN	O	O
CLSM	NN	O	O
)	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
impermeable	NN	O	O
testosterone-BSA	NN	O	B-protein
conjugate	NN	O	I-protein
induces	NN	O	O
a	NN	O	O
rapid	NN	O	O
rise	NN	O	O
(	NN	O	O
<	NN	O	O
5	NN	O	O
s	NN	O	O
)	NN	O	O
in	NN	O	O
[	NN	O	O
Ca2+	NN	O	O
]	NN	O	O
i	NN	O	O
of	NN	O	O
Fura-2-loaded	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
rise	NN	O	O
reflects	NN	O	O
influx	NN	O	O
of	NN	O	O
extracellular	NN	O	O
Ca2+	NN	O	O
through	NN	O	O
non-voltage-gated	NN	O	O
and	NN	O	O
Ni2+-blockable	NN	O	B-protein
Ca2+	NN	O	I-protein
channels	NN	O	I-protein
of	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
.	NN	O	O

The	NN	O	O
testosterone-BSA-induced	NN	O	O
Ca2+	NN	O	O
import	NN	O	O
is	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
cyproterone	NN	O	O
,	NN	O	O
a	NN	O	O
blocker	NN	O	O
of	NN	O	O
the	NN	O	O
AR	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
AR	NN	O	B-protein
are	NN	O	O
not	NN	O	O
detectable	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
intact	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
when	NN	O	O
using	NN	O	O
anti-AR	NN	O	B-protein
antibodies	NN	O	I-protein
directed	NN	O	O
against	NN	O	O
the	NN	O	O
amino	NN	O	B-protein
and	NN	O	I-protein
carboxy	NN	O	I-protein
terminus	NN	O	I-protein
of	NN	O	O
the	NN	O	O
AR	NN	O	B-protein
,	NN	O	O
although	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
contain	NN	O	O
AR	NN	O	B-protein
,	NN	O	O
as	NN	O	O
revealed	NN	O	O
by	NN	O	O
reverse	NN	O	O
transcription-polymerase	NN	O	O
chain	NN	O	O
reactions	NN	O	O
and	NN	O	O
Western	NN	O	O
blotting	NN	O	O
.	NN	O	O

AR	NN	O	B-protein
can	NN	O	O
be	NN	O	O
visualized	NN	O	O
with	NN	O	O
the	NN	O	O
anti-AR	NN	O	B-protein
antibodies	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
permeabilized	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
using	NN	O	O
CLSM	NN	O	O
,	NN	O	O
though	NN	O	O
AR	NN	O	B-protein
are	NN	O	O
not	NN	O	O
detectable	NN	O	O
in	NN	O	O
cytosol	NN	O	O
fractions	NN	O	O
when	NN	O	O
using	NN	O	O
the	NN	O	O
charcoal	NN	O	O
binding	NN	O	O
assay	NN	O	O
with	NN	O	O
3H-R1881	NN	O	O
as	NN	O	O
ligand	NN	O	O
.	NN	O	O

Cytoplasmic	NN	O	B-protein
AR	NN	O	I-protein
do	NN	O	O
not	NN	O	O
translocate	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
testosterone	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
cytoplasmic	NN	O	B-protein
AR	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
cancer	NN	O	I-cell_line
LNCaP	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
classical	NN	O	B-protein
AR	NN	O	I-protein
present	NN	O	O
in	NN	O	O
splenic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
not	NN	O	O
active	NN	O	O
in	NN	O	O
the	NN	O	O
genomic	NN	O	O
pathway	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
for	NN	O	O
testosterone	NN	O	O
are	NN	O	O
in	NN	O	O
a	NN	O	O
functionally	NN	O	O
active	NN	O	O
state	NN	O	O
,	NN	O	O
enabling	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
a	NN	O	O
nongenomic	NN	O	O
response	NN	O	O
to	NN	O	O
testosterone	NN	O	O
.	NN	O	O

-DOCSTART-	O

Limited	NN	O	O
proteolysis	NN	O	O
for	NN	O	O
assaying	NN	O	O
ligand	NN	O	O
binding	NN	O	O
affinities	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
natural	NN	O	O
or	NN	O	O
synthetic	NN	O	O
ligands	NN	O	O
to	NN	O	O
nuclear	NN	O	B-protein
receptors	NN	O	I-protein
is	NN	O	O
the	NN	O	O
triggering	NN	O	O
event	NN	O	O
leading	NN	O	O
to	NN	O	O
gene	NN	O	O
transcription	NN	O	O
activation	NN	O	O
or	NN	O	O
repression	NN	O	O
.	NN	O	O

Ligand	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
ligand	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
these	NN	O	O
receptors	NN	O	B-protein
induces	NN	O	O
conformational	NN	O	O
changes	NN	O	O
that	NN	O	O
are	NN	O	O
evidenced	NN	O	O
by	NN	O	O
an	NN	O	O
increased	NN	O	O
resistance	NN	O	O
of	NN	O	O
this	NN	O	O
domain	NN	O	O
to	NN	O	O
proteases	NN	O	B-protein
.	NN	O	O

In	NN	O	B-protein
vitro	NN	O	I-protein
labeled	NN	O	I-protein
receptors	NN	O	I-protein
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
various	NN	O	O
synthetic	NN	O	O
or	NN	O	O
natural	NN	O	O
agonists	NN	O	O
or	NN	O	O
antagonists	NN	O	O
and	NN	O	O
submitted	NN	O	O
to	NN	O	O
trypsin	NN	O	B-protein
digestion	NN	O	O
.	NN	O	O

Proteolysis	NN	O	O
products	NN	O	O
were	NN	O	O
separated	NN	O	O
by	NN	O	O
SDS-PAGE	NN	O	O
and	NN	O	O
quantified	NN	O	O
.	NN	O	O

The	NN	O	O
amount	NN	O	O
of	NN	O	O
trypsin	NN	O	B-protein
-resistant	NN	O	O
fragments	NN	O	O
was	NN	O	O
proportional	NN	O	O
to	NN	O	O
receptor	NN	O	O
occupancy	NN	O	O
by	NN	O	O
the	NN	O	O
ligand	NN	O	O
,	NN	O	O
and	NN	O	O
allowed	NN	O	O
the	NN	O	O
determination	NN	O	O
of	NN	O	O
dissociation	NN	O	O
constants	NN	O	O
(	NN	O	O
kDa	NN	O	O
)	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
or	NN	O	I-protein
mutated	NN	O	I-protein
human	NN	O	I-protein
retinoic	NN	O	I-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
as	NN	O	O
a	NN	O	O
model	NN	O	O
,	NN	O	O
kDa	NN	O	O
values	NN	O	O
determined	NN	O	O
by	NN	O	O
classical	NN	O	O
competition	NN	O	O
binding	NN	O	O
assays	NN	O	O
using	NN	O	O
tritiated	NN	O	O
ligands	NN	O	O
are	NN	O	O
in	NN	O	O
agreement	NN	O	O
with	NN	O	O
those	NN	O	O
measured	NN	O	O
by	NN	O	O
the	NN	O	O
proteolytic	NN	O	O
assay	NN	O	O
.	NN	O	O

This	NN	O	O
method	NN	O	O
was	NN	O	O
successfully	NN	O	O
extended	NN	O	O
to	NN	O	O
human	NN	O	B-protein
retinoic	NN	O	I-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
thus	NN	O	O
providing	NN	O	O
a	NN	O	O
basis	NN	O	O
for	NN	O	O
a	NN	O	O
new	NN	O	O
,	NN	O	O
faster	NN	O	O
assay	NN	O	O
to	NN	O	O
determine	NN	O	O
simultaneously	NN	O	O
the	NN	O	O
affinity	NN	O	O
and	NN	O	O
conformation	NN	O	O
of	NN	O	O
receptors	NN	O	O
when	NN	O	O
bound	NN	O	O
to	NN	O	O
a	NN	O	O
given	NN	O	O
ligand	NN	O	O
.	NN	O	O

-DOCSTART-	O

Genes	NN	O	O
that	NN	O	O
regulate	NN	O	O
interleukin-4	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Interleukin-4	NN	O	B-protein
is	NN	O	O
an	NN	O	O
immunomodulatory	NN	O	B-protein
cytokine	NN	O	I-protein
which	NN	O	O
plays	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
allergic	NN	O	O
and	NN	O	O
atopic	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

Significant	NN	O	O
progress	NN	O	O
has	NN	O	O
been	NN	O	O
made	NN	O	O
in	NN	O	O
gaining	NN	O	O
a	NN	O	O
detailed	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-4	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
recent	NN	O	O
identification	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
several	NN	O	O
key	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
has	NN	O	O
helped	NN	O	O
to	NN	O	O
elucidate	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
T	NN	O	B-DNA
helper	NN	O	I-DNA
cell	NN	O	I-DNA
cytokine	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
contain	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
STAT3	NN	O	B-protein
constitutively	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
serine	NN	O	O
residues	NN	O	O
.	NN	O	O

To	NN	O	O
explore	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CLL	NN	O	O
)	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
whether	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
factors	NN	O	I-protein
was	NN	O	O
abnormal	NN	O	O
in	NN	O	O
cells	NN	O	O
from	NN	O	O
CLL	NN	O	O
patients	NN	O	O
.	NN	O	O

No	NN	O	O
constitutive	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
was	NN	O	O
detected	NN	O	O
on	NN	O	O
any	NN	O	O
STAT	NN	O	B-protein
in	NN	O	O
CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
assess	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
serine	NN	O	O
residues	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
in	NN	O	O
CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
raised	NN	O	O
antibodies	NN	O	O
that	NN	O	O
specifically	NN	O	O
recognize	NN	O	O
the	NN	O	O
form	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
phosphorylated	NN	O	O
on	NN	O	O
ser-727	NN	O	O
and	NN	O	O
the	NN	O	O
form	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
phosphorylated	NN	O	O
on	NN	O	O
ser-727	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
in	NN	O	O
100	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
CLL	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
32	NN	O	O
)	NN	O	O
,	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
were	NN	O	O
constitutively	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
serine	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
normal	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
or	NN	O	O
CD5+	NN	O	B-cell_type
)	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
tonsils	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
this	NN	O	O
phosphorylation	NN	O	O
was	NN	O	O
absent	NN	O	O
.	NN	O	O

Serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
was	NN	O	O
seen	NN	O	O
occasionally	NN	O	O
in	NN	O	O
other	NN	O	O
leukemias	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
was	NN	O	O
a	NN	O	O
universal	NN	O	O
finding	NN	O	O
only	NN	O	O
in	NN	O	O
CLL	NN	O	O
.	NN	O	O

The	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
these	NN	O	O
STATs	NN	O	B-protein
was	NN	O	O
a	NN	O	O
continuous	NN	O	O
process	NN	O	O
,	NN	O	O
as	NN	O	O
incubation	NN	O	O
of	NN	O	O
CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
kinase	NN	O	B-protein
inhibitor	NN	O	I-protein
H7	NN	O	I-protein
led	NN	O	O
to	NN	O	O
the	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
these	NN	O	O
serine	NN	O	O
residues	NN	O	O
.	NN	O	O

The	NN	O	O
STAT	NN	O	O
serine	NN	O	O
kinase	NN	O	O
in	NN	O	O
CLL	NN	O	O
cells	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
identified	NN	O	O
,	NN	O	O
and	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
neither	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
nor	NN	O	O
pp70	NN	O	B-protein
(	NN	O	I-protein
s6k	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
the	NN	O	O
constitutive	NN	O	O
serine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
is	NN	O	O
present	NN	O	O
in	NN	O	O
all	NN	O	O
CLL	NN	O	O
samples	NN	O	O
tested	NN	O	O
to	NN	O	O
date	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
physiologic	NN	O	O
significance	NN	O	O
of	NN	O	O
this	NN	O	O
modification	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
determined	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
expression	NN	O	O
by	NN	O	O
Gardnerella	NN	O	O
vaginalis	NN	O	O
.	NN	O	O

Bacterial	NN	O	O
vaginosis	NN	O	O
(	NN	O	O
BV	NN	O	O
)	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
rate	NN	O	O
of	NN	O	O
sexual	NN	O	O
transmission	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
type	NN	O	O
1	NN	O	O
,	NN	O	O
and	NN	O	O
Gardnerella	NN	O	O
vaginalis	NN	O	O
is	NN	O	O
frequently	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
genital	NN	O	O
tracts	NN	O	O
of	NN	O	O
women	NN	O	O
with	NN	O	O
BV	NN	O	O
.	NN	O	O

G.	NN	O	O
vaginalis	NN	O	O
lysates	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
significantly	NN	O	O
stimulate	NN	O	O
HIV	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Stimulation	NN	O	O
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
when	NN	O	O
lysates	NN	O	O
were	NN	O	O
heated	NN	O	O
at	NN	O	O
100	NN	O	O
degrees	NN	O	O
C	NN	O	O
for	NN	O	O
5	NN	O	O
min	NN	O	O
but	NN	O	O
was	NN	O	O
reduced	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
lysozyme	NN	O	B-protein
or	NN	O	O
protease	NN	O	B-protein
.	NN	O	O

G.	NN	O	O
vaginalis	NN	O	O
lysates	NN	O	O
also	NN	O	O
activated	NN	O	O
HIV	NN	O	O
expression	NN	O	O
in	NN	O	O
certain	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

G.	NN	O	O
vaginalis	NN	O	O
lysates	NN	O	O
activated	NN	O	O
HIV	NN	O	O
long-terminal	NN	O	O
repeat	NN	O	O
transcription	NN	O	O
in	NN	O	O
HIV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
increased	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
indicating	NN	O	O
an	NN	O	O
effect	NN	O	O
by	NN	O	O
G.	NN	O	O
vaginalis	NN	O	O
on	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV	NN	O	O
production	NN	O	O
by	NN	O	O
G.	NN	O	O
vaginalis	NN	O	O
suggests	NN	O	O
that	NN	O	O
genital	NN	O	O
tract	NN	O	O
infection	NN	O	O
with	NN	O	O
G.	NN	O	O
vaginalis	NN	O	O
increases	NN	O	O
the	NN	O	O
risk	NN	O	O
of	NN	O	O
HIV	NN	O	O
transmission	NN	O	O
by	NN	O	O
increasing	NN	O	O
HIV	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
genital	NN	O	O
tract	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
explain	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
the	NN	O	O
increased	NN	O	O
rate	NN	O	O
of	NN	O	O
HIV	NN	O	O
transmission	NN	O	O
in	NN	O	O
women	NN	O	O
with	NN	O	O
BV	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin-10	NN	O	B-protein
inhibits	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
interferon	NN	O	B-DNA
alpha-	NN	O	I-DNA
and	NN	O	I-DNA
interferon	NN	O	I-DNA
gamma-	NN	O	I-DNA
induced	NN	O	I-DNA
genes	NN	O	I-DNA
by	NN	O	O
suppressing	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
.	NN	O	O

Interleukin-10	NN	O	B-protein
(	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
helps	NN	O	O
maintain	NN	O	O
polarized	NN	O	B-cell_type
T-helper	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
a	NN	O	O
T-helper	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
2	NN	O	I-cell_type
(	NN	O	I-cell_type
Th2	NN	O	I-cell_type
)	NN	O	I-cell_type
phenotype	NN	O	I-cell_type
.	NN	O	O

Part	NN	O	O
of	NN	O	O
this	NN	O	O
process	NN	O	O
involves	NN	O	O
the	NN	O	O
prevention	NN	O	O
of	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
are	NN	O	O
a	NN	O	O
primary	NN	O	O
source	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
(	NN	O	O
IFNgamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
activator	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
Th2	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Because	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	O
are	NN	O	O
important	NN	O	O
mediators	NN	O	O
of	NN	O	O
Th1-type	NN	O	O
responses	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
delayed-type	NN	O	O
hypersensitivity	NN	O	O
,	NN	O	O
we	NN	O	O
sought	NN	O	O
to	NN	O	O
determine	NN	O	O
if	NN	O	O
IL-10	NN	O	B-protein
could	NN	O	O
directly	NN	O	O
mediate	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IFNgamma-	NN	O	O
and	NN	O	O
IFNalpha-induced	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Highly	NN	O	O
purified	NN	O	O
monocytes	NN	O	B-cell_type
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
IL-10	NN	O	B-protein
for	NN	O	O
60	NN	O	O
to	NN	O	O
90	NN	O	O
minutes	NN	O	O
before	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
IFNgamma	NN	O	B-protein
or	NN	O	O
IFNalpha	NN	O	B-protein
.	NN	O	O

IL-10	NN	O	B-protein
preincubation	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
for	NN	O	O
several	NN	O	O
IFN-induced	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
IP-10	NN	O	B-protein
,	NN	O	O
ISG54	NN	O	B-protein
,	NN	O	O
and	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
.	NN	O	O

The	NN	O	O
reduction	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
resulted	NN	O	O
from	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
to	NN	O	O
suppress	NN	O	O
IFN	NN	O	B-protein
-induced	NN	O	O
assembly	NN	O	O
of	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
factors	NN	O	I-protein
to	NN	O	O
specific	NN	O	B-DNA
promoter	NN	O	I-DNA
motifs	NN	O	I-DNA
on	NN	O	O
IFNalpha-	NN	O	B-DNA
and	NN	O	I-DNA
IFNgamma-inducible	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
was	NN	O	O
accomplished	NN	O	O
by	NN	O	O
preventing	NN	O	O
the	NN	O	O
IFN-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
both	NN	O	O
IFNalpha-	NN	O	B-protein
and	NN	O	I-protein
IFNgamma-induced	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
can	NN	O	O
directly	NN	O	O
inhibit	NN	O	O
STAT-dependent	NN	O	B-DNA
early	NN	O	I-DNA
response	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
both	NN	O	O
IFNalpha	NN	O	B-protein
and	NN	O	O
IFNgamma	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
by	NN	O	O
suppressing	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
.	NN	O	O

This	NN	O	O
may	NN	O	O
occur	NN	O	O
through	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
suppressor	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
signaling	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
SOCS3	NN	O	B-protein
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Clonality	NN	O	O
analysis	NN	O	O
of	NN	O	O
refractory	NN	O	O
anemia	NN	O	O
with	NN	O	O
ring	NN	O	B-cell_type
sideroblasts	NN	O	I-cell_type
:	NN	O	O
simultaneous	NN	O	O
study	NN	O	O
of	NN	O	O
clonality	NN	O	O
and	NN	O	O
cytochemistry	NN	O	O
of	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

X	NN	O	O
chromosome	NN	O	O
inactivation	NN	O	O
and	NN	O	O
polymorphism	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
androgen	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
HUMARA	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
applied	NN	O	O
for	NN	O	O
analyzing	NN	O	O
the	NN	O	O
clonality	NN	O	O
of	NN	O	O
blood	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
clonal	NN	O	O
relationship	NN	O	O
was	NN	O	O
investigated	NN	O	O
between	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
polymorphonuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PMNCs	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
marrow	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
ringed	NN	O	B-cell_type
sideroblasts	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
refractory	NN	O	O
anemia	NN	O	O
with	NN	O	O
ring	NN	O	B-cell_type
sideroblasts	NN	O	I-cell_type
(	NN	O	O
RARS	NN	O	B-cell_type
)	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
of	NN	O	O
HUMARA	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
X-inactivation	NN	O	O
patterns	NN	O	O
of	NN	O	O
circulating	NN	O	O
PMNCs	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
individual	NN	O	O
granulocyte	NN	O	B-cell_line
colonies	NN	O	I-cell_line
grown	NN	O	O
in	NN	O	O
vitro	NN	O	O
from	NN	O	O
bone	NN	O	O
marrow	NN	O	O
cells	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

The	NN	O	O
development	NN	O	O
of	NN	O	O
ringed	NN	O	B-cell_type
sideroblasts	NN	O	I-cell_type
in	NN	O	O
erythroid	NN	O	B-cell_line
colonies	NN	O	I-cell_line
by	NN	O	O
iron	NN	O	O
staining	NN	O	O
and	NN	O	O
their	NN	O	O
X-inactivation	NN	O	O
pattern	NN	O	O
were	NN	O	O
also	NN	O	O
examined	NN	O	O
.	NN	O	O

All	NN	O	O
three	NN	O	O
RARS	NN	O	B-cell_type
patients	NN	O	O
showed	NN	O	O
monoclonal	NN	O	O
PMNCs	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
granulocyte	NN	O	B-cell_line
colonies	NN	O	I-cell_line
,	NN	O	O
however	NN	O	O
,	NN	O	O
two	NN	O	O
different	NN	O	O
X-inactivation	NN	O	O
patterns	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
all	NN	O	O
patients	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
non-clonal	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
remained	NN	O	O
in	NN	O	O
the	NN	O	O
bone	NN	O	O
marrow	NN	O	O
.	NN	O	O

All	NN	O	O
erythroid	NN	O	B-cell_line
colonies	NN	O	I-cell_line
consisted	NN	O	O
of	NN	O	O
ringed	NN	O	B-cell_type
sideroblasts	NN	O	I-cell_type
exclusively	NN	O	O
showed	NN	O	O
one	NN	O	O
pattern	NN	O	O
dominant	NN	O	O
in	NN	O	O
those	NN	O	O
of	NN	O	O
PMNCs	NN	O	B-cell_type
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
non-clonal	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
persist	NN	O	O
in	NN	O	O
some	NN	O	O
RARS	NN	O	B-cell_type
cases	NN	O	O
,	NN	O	O
that	NN	O	O
erythroid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
show	NN	O	O
mosaicism	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
ringed	NN	O	B-cell_type
sideroblasts	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
derived	NN	O	O
from	NN	O	O
an	NN	O	O
abnormal	NN	O	O
clone	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
this	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Granulosa	NN	O	O
cell	NN	O	O
tumor	NN	O	O
of	NN	O	O
the	NN	O	O
ovary	NN	O	O
.	NN	O	O

Immunohistochemical	NN	O	O
evidence	NN	O	O
of	NN	O	O
low	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
and	NN	O	O
virtual	NN	O	O
absence	NN	O	O
of	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
p53	NN	O	B-DNA
tumor-suppressor	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

BACKGROUND	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
Because	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
immunohistochemistry	NN	O	O
in	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
granulosa	NN	O	O
cell	NN	O	O
tumor	NN	O	O
(	NN	O	O
GCT	NN	O	O
)	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
fully	NN	O	O
explored	NN	O	O
,	NN	O	O
routinely	NN	O	O
processed	NN	O	O
(	NN	O	O
formalin-fixed	NN	O	O
,	NN	O	O
paraffin-embedded	NN	O	O
)	NN	O	O
tissue	NN	O	O
from	NN	O	O
11	NN	O	O
GCT	NN	O	O
,	NN	O	O
adult	NN	O	O
type	NN	O	O
,	NN	O	O
was	NN	O	O
investigated	NN	O	O
immunohistochemically	NN	O	O
(	NN	O	O
ABC	NN	O	O
method	NN	O	O
)	NN	O	O
with	NN	O	O
a	NN	O	O
broad	NN	O	O
spectrum	NN	O	O
of	NN	O	O
antibodies	NN	O	O
against	NN	O	O
various	NN	O	O
markers	NN	O	O
,	NN	O	O
including	NN	O	O
p53	NN	O	B-protein
and	NN	O	O
Ki-67	NN	O	B-protein
.	NN	O	O

All	NN	O	O
of	NN	O	O
the	NN	O	O
tumors	NN	O	O
exhibited	NN	O	O
typical	NN	O	O
morphology	NN	O	O
,	NN	O	O
were	NN	O	O
limited	NN	O	O
to	NN	O	O
the	NN	O	O
ovary	NN	O	O
(	NN	O	O
stage	NN	O	O
I	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
7	NN	O	O
cases	NN	O	O
followed	NN	O	O
a	NN	O	O
benign	NN	O	O
clinical	NN	O	O
course	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
All	NN	O	O
the	NN	O	O
tumors	NN	O	O
exhibited	NN	O	O
strong	NN	O	O
expression	NN	O	O
of	NN	O	O
vimentin	NN	O	B-protein
,	NN	O	O
but	NN	O	O
most	NN	O	O
other	NN	O	O
antigens	NN	O	O
(	NN	O	O
including	NN	O	O
smooth	NN	O	B-protein
muscle	NN	O	I-protein
actin	NN	O	I-protein
)	NN	O	O
were	NN	O	O
expressed	NN	O	O
infrequently	NN	O	O
by	NN	O	O
a	NN	O	O
minority	NN	O	O
of	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
not	NN	O	O
at	NN	O	O
all	NN	O	O
.	NN	O	O

Tumor	NN	O	O
cells	NN	O	O
in	NN	O	O
9	NN	O	O
GCT	NN	O	O
expressed	NN	O	O
inhibin	NN	O	O
A	NN	O	O
.	NN	O	O

All	NN	O	O
the	NN	O	O
tumors	NN	O	O
exhibited	NN	O	O
very	NN	O	O
low	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
,	NN	O	O
fewer	NN	O	O
than	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	O
cell	NN	O	O
nuclei	NN	O	O
being	NN	O	O
stained	NN	O	O
by	NN	O	O
the	NN	O	O
antibody	NN	O	B-protein
MIB-1	NN	O	I-protein
(	NN	O	O
Ki-67	NN	O	B-protein
antigen	NN	O	I-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
antibody	NN	O	B-protein
D07	NN	O	I-protein
revealed	NN	O	O
marked	NN	O	O
overexpression	NN	O	O
of	NN	O	O
p53	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
only	NN	O	O
one	NN	O	O
tumor	NN	O	O
.	NN	O	O

Clinical	NN	O	O
outcome	NN	O	O
was	NN	O	O
not	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
immunophenotypic	NN	O	O
differences	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
The	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
GCT	NN	O	O
should	NN	O	O
be	NN	O	O
based	NN	O	O
primarily	NN	O	O
on	NN	O	O
the	NN	O	O
typical	NN	O	O
morphology	NN	O	O
revealed	NN	O	O
by	NN	O	O
conventional	NN	O	O
stains	NN	O	O
,	NN	O	O
but	NN	O	O
additional	NN	O	O
immunohistochemical	NN	O	O
staining	NN	O	O
with	NN	O	O
a	NN	O	O
small	NN	O	O
panel	NN	O	O
of	NN	O	O
selected	NN	O	O
antibodies	NN	O	O
(	NN	O	O
for	NN	O	O
example	NN	O	O
,	NN	O	O
against	NN	O	O
keratin	NN	O	B-protein
,	NN	O	O
vimentin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
inhibin	NN	O	B-protein
A	NN	O	I-protein
)	NN	O	O
may	NN	O	O
be	NN	O	O
helpful	NN	O	O
in	NN	O	O
a	NN	O	O
few	NN	O	O
cases	NN	O	O
.	NN	O	O

The	NN	O	O
very	NN	O	O
low	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
and	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
overexpression	NN	O	O
of	NN	O	O
p53	NN	O	B-protein
protein	NN	O	I-protein
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
benign	NN	O	O
clinical	NN	O	O
behavior	NN	O	O
of	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
GCT	NN	O	O
.	NN	O	O

-DOCSTART-	O

Protective	NN	O	O
effects	NN	O	O
of	NN	O	O
notch-1	NN	O	B-protein
on	NN	O	O
TCR	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
Notch	NN	O	B-protein
receptor	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
originally	NN	O	O
identified	NN	O	O
in	NN	O	O
Drosophila	NN	O	O
and	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
mediate	NN	O	O
cell	NN	O	O
to	NN	O	O
cell	NN	O	O
communication	NN	O	O
and	NN	O	O
influence	NN	O	O
cell	NN	O	O
fate	NN	O	O
decisions	NN	O	O
.	NN	O	O

Members	NN	O	O
of	NN	O	O
this	NN	O	O
family	NN	O	O
have	NN	O	O
been	NN	O	O
isolated	NN	O	O
from	NN	O	O
invertebrates	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
vertebrates	NN	O	O
.	NN	O	O

We	NN	O	O
isolated	NN	O	O
mouse	NN	O	B-protein
Notch-1	NN	O	I-protein
in	NN	O	O
a	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
screen	NN	O	O
with	NN	O	O
Nur77	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
protein	NN	O	O
that	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
previously	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
data	NN	O	O
presented	NN	O	O
below	NN	O	O
indicate	NN	O	O
that	NN	O	O
Notch-1	NN	O	B-protein
expression	NN	O	O
provides	NN	O	O
significant	NN	O	O
protection	NN	O	O
to	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
from	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
new	NN	O	O
antiapoptotic	NN	O	O
role	NN	O	O
for	NN	O	O
Notch-1	NN	O	B-protein
,	NN	O	O
providing	NN	O	O
evidence	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
regulating	NN	O	O
cell	NN	O	O
fate	NN	O	O
decisions	NN	O	O
,	NN	O	O
Notch-1	NN	O	B-protein
can	NN	O	O
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
controlling	NN	O	O
levels	NN	O	O
of	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Comparison	NN	O	O
of	NN	O	O
primary	NN	O	O
sensitization	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
varicella-zoster	NN	O	O
virus	NN	O	O
peptides	NN	O	O
by	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
responses	NN	O	O
elicited	NN	O	O
in	NN	O	O
vivo	NN	O	O
by	NN	O	O
varicella	NN	O	O
vaccination	NN	O	O
.	NN	O	O

Dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
DC	NN	O	B-cell_type
)	NN	O	O
are	NN	O	O
potent	NN	O	O
APC	NN	O	B-cell_type
during	NN	O	O
primary	NN	O	O
and	NN	O	O
secondary	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
objective	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
human	NN	O	B-cell_type
DC	NN	O	I-cell_type
mediate	NN	O	O
in	NN	O	O
vitro	NN	O	O
sensitization	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
epitopes	NN	O	B-protein
of	NN	O	O
the	NN	O	O
immediate	NN	O	B-protein
early	NN	O	I-protein
62	NN	O	I-protein
(	NN	O	I-protein
IE62	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
varicella	NN	O	O
zoster	NN	O	O
virus	NN	O	O
(	NN	O	O
VZV	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
CD4+	NN	O	B-protein
T	NN	O	O
cell	NN	O	O
proliferative	NN	O	O
responses	NN	O	O
to	NN	O	O
eight	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
representing	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequences	NN	O	O
of	NN	O	O
the	NN	O	O
VZV	NN	O	B-protein
IE62	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
assessed	NN	O	O
using	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
DC	NN	O	B-cell_type
from	NN	O	O
VZV-susceptible	NN	O	O
donors	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
objective	NN	O	O
was	NN	O	O
to	NN	O	O
compare	NN	O	O
in	NN	O	O
vitro	NN	O	O
responses	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
responses	NN	O	O
to	NN	O	O
VZV	NN	O	O
peptides	NN	O	O
induced	NN	O	O
in	NN	O	O
vivo	NN	O	O
after	NN	O	O
immunization	NN	O	O
with	NN	O	O
varicella	NN	O	O
vaccine	NN	O	O
.	NN	O	O

T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
three	NN	O	O
peptides	NN	O	O
,	NN	O	O
P1	NN	O	O
,	NN	O	O
P4	NN	O	O
,	NN	O	O
and	NN	O	O
P7	NN	O	O
,	NN	O	O
in	NN	O	O
71-100	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
donors	NN	O	O
tested	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
vaccination	NN	O	O
using	NN	O	O
DC	NN	O	B-cell_type
as	NN	O	O
APC	NN	O	B-cell_type
.	NN	O	O

Monocytes	NN	O	B-cell_type
were	NN	O	O
effective	NN	O	O
APC	NN	O	B-cell_type
for	NN	O	O
VZV	NN	O	O
peptides	NN	O	O
only	NN	O	O
after	NN	O	O
immunization	NN	O	O
.	NN	O	O

Two	NN	O	O
peptides	NN	O	O
,	NN	O	O
P2	NN	O	O
and	NN	O	O
P8	NN	O	O
,	NN	O	O
induced	NN	O	O
naive	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
less	NN	O	O
effectively	NN	O	O
and	NN	O	O
were	NN	O	O
also	NN	O	O
less	NN	O	O
immunogenic	NN	O	O
for	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
vaccinated	NN	O	O
or	NN	O	O
naturally	NN	O	O
immune	NN	O	O
donors	NN	O	O
.	NN	O	O

T	NN	O	O
cell	NN	O	O
recognition	NN	O	O
of	NN	O	O
specific	NN	O	O
peptides	NN	O	O
was	NN	O	O
concordant	NN	O	O
between	NN	O	O
naive	NN	O	O
,	NN	O	O
DC	NN	O	B-cell_type
-mediated	NN	O	O
responses	NN	O	O
,	NN	O	O
and	NN	O	O
postvaccine	NN	O	O
responses	NN	O	O
using	NN	O	O
monocytes	NN	O	B-cell_type
as	NN	O	O
APC	NN	O	B-cell_type
in	NN	O	O
69	NN	O	O
%	NN	O	O
of	NN	O	O
comparisons	NN	O	O
(	NN	O	O
p	NN	O	O
=	NN	O	O
0.05	NN	O	O
;	NN	O	O
chi2	NN	O	O
)	NN	O	O
;	NN	O	O
the	NN	O	O
predictive	NN	O	O
value	NN	O	O
of	NN	O	O
a	NN	O	O
positive	NN	O	O
response	NN	O	O
to	NN	O	O
an	NN	O	O
IE62	NN	O	O
peptide	NN	O	O
before	NN	O	O
immunization	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
sensitization	NN	O	O
in	NN	O	O
vivo	NN	O	O
was	NN	O	O
82	NN	O	O
%	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
indicate	NN	O	O
that	NN	O	O
primary	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
detected	NN	O	O
in	NN	O	O
vitro	NN	O	O
using	NN	O	O
DC	NN	O	B-cell_type
as	NN	O	O
APC	NN	O	B-cell_type
may	NN	O	O
be	NN	O	O
useful	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
potential	NN	O	O
immunogenicity	NN	O	O
of	NN	O	O
viral	NN	O	O
protein	NN	O	O
epitopes	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mechanism	NN	O	O
of	NN	O	O
interleukin-10	NN	O	B-protein
inhibition	NN	O	O
of	NN	O	O
T-helper	NN	O	O
cell	NN	O	O
activation	NN	O	O
by	NN	O	O
superantigen	NN	O	B-protein
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
interleukin-10	NN	O	B-protein
(	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
entry	NN	O	O
of	NN	O	O
quiescent	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
the	NN	O	O
superantigen	NN	O	B-protein
staphylococcal	NN	O	B-protein
enterotoxin	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
SEB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
arrested	NN	O	O
cells	NN	O	O
at	NN	O	O
G0/G1	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
treatment	NN	O	O
prevented	NN	O	O
the	NN	O	O
downregulation	NN	O	O
of	NN	O	O
p27	NN	O	B-protein
(	NN	O	I-protein
Kip1	NN	O	I-protein
)	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitory	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
controls	NN	O	O
progression	NN	O	O
out	NN	O	O
of	NN	O	O
the	NN	O	O
G0	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
also	NN	O	O
prevented	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
the	NN	O	O
G1	NN	O	B-protein
cyclins	NN	O	I-protein
D2	NN	O	B-protein
and	NN	O	O
D3	NN	O	B-protein
,	NN	O	O
proteins	NN	O	O
necessary	NN	O	O
for	NN	O	O
entry	NN	O	O
and	NN	O	O
progression	NN	O	O
through	NN	O	O
the	NN	O	O
G1	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

Associated	NN	O	O
with	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
suppressed	NN	O	O
SEB	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
exogenous	NN	O	O
IL-2	NN	O	B-protein
to	NN	O	O
IL-10-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
significantly	NN	O	O
reversed	NN	O	O
the	NN	O	O
antiproliferative	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
effects	NN	O	O
on	NN	O	O
the	NN	O	O
early	NN	O	B-protein
G1	NN	O	I-protein
proteins	NN	O	I-protein
p27	NN	O	B-protein
(	NN	O	I-protein
Kip1	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
cyclin	NN	O	B-protein
D2	NN	O	I-protein
were	NN	O	O
similarly	NN	O	O
reversed	NN	O	O
by	NN	O	O
exogenous	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
this	NN	O	O
reversal	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
was	NN	O	O
pronounced	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
not	NN	O	O
complete	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
may	NN	O	O
have	NN	O	O
some	NN	O	O
effects	NN	O	O
not	NN	O	O
directly	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Cell	NN	O	O
separation	NN	O	O
experiments	NN	O	O
suggest	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
can	NN	O	O
effect	NN	O	O
purified	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
directly	NN	O	O
,	NN	O	O
providing	NN	O	O
functional	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
receptors	NN	O	O
on	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

IL-10	NN	O	B-protein
also	NN	O	O
inhibited	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
transcriptional	NN	O	I-protein
regulators	NN	O	I-protein
c-fos	NN	O	B-protein
and	NN	O	O
c-jun	NN	O	B-protein
,	NN	O	O
which	NN	O	O
also	NN	O	O
inhibit	NN	O	O
other	NN	O	O
cell	NN	O	O
functions	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
regulation	NN	O	O
of	NN	O	O
quiescent	NN	O	O
CD4	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
is	NN	O	O
mainly	NN	O	O
by	NN	O	O
blocking	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
that	NN	O	O
is	NN	O	O
critical	NN	O	O
to	NN	O	O
downregulation	NN	O	O
of	NN	O	O
p27	NN	O	B-protein
(	NN	O	I-protein
Kip1	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
upregulation	NN	O	O
of	NN	O	O
D	NN	O	B-protein
cyclins	NN	O	I-protein
in	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
and	NN	O	O
entry	NN	O	O
into	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

-DOCSTART-	O

IFN-gamma	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
inhibit	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-1	NN	O	O
receptor	NN	O	O
type	NN	O	O
I	NN	O	O
and	NN	O	O
type	NN	O	O
II	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
in	NN	O	O
human	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
Th2-type	NN	O	B-protein
cytokines	NN	O	I-protein
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
distinct	NN	O	O
subset	NN	O	O
of	NN	O	O
genes	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
include	NN	O	O
Fc	NN	O	B-protein
epsilonRII	NN	O	I-protein
(	NN	O	O
CD23	NN	O	B-protein
)	NN	O	O
,	NN	O	O
15-lipoxygenase	NN	O	B-protein
,	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	I-protein
antagonist	NN	O	I-protein
(	NN	O	O
IL-1ra	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
IL-1	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
IL-1R	NN	O	B-protein
)	NN	O	O
.	NN	O	O

IFN-gamma	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
inhibit	NN	O	O
induction	NN	O	O
of	NN	O	O
CD23	NN	O	B-protein
and	NN	O	O
15-lipoxygenase	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
on	NN	O	O
type	NN	O	O
I	NN	O	O
and	NN	O	O
type	NN	O	O
II	NN	O	O
IL-1R	NN	O	O
gene	NN	O	O
expression	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
defined	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
on	NN	O	O
both	NN	O	O
basal	NN	O	O
and	NN	O	O
IL-4	NN	O	B-protein
/	NN	O	O
IL-13	NN	O	B-protein
-induced	NN	O	O
IL-1R	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
induced	NN	O	O
dose-	NN	O	O
and	NN	O	O
time-dependent	NN	O	O
increases	NN	O	O
in	NN	O	O
IL-1RI	NN	O	O
and	NN	O	O
IL-1RII	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

IFN-gamma	NN	O	B-protein
decreased	NN	O	O
basal	NN	O	O
expression	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
IL-1RI	NN	O	B-protein
and	NN	O	O
IL-1RII	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
was	NN	O	O
transcriptionally	NN	O	O
mediated	NN	O	O
,	NN	O	O
and	NN	O	O
correlated	NN	O	O
directly	NN	O	O
with	NN	O	O
decreased	NN	O	O
production	NN	O	O
of	NN	O	O
soluble	NN	O	O
IL-1RII	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
suppress	NN	O	O
IL-1RI	NN	O	O
and	NN	O	O
IL-1RII	NN	O	B-protein
mRNA	NN	O	O
levels	NN	O	O
was	NN	O	O
not	NN	O	O
unique	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
because	NN	O	O
IL-10	NN	O	B-protein
also	NN	O	O
inhibited	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
in	NN	O	O
IL-4/IL-13-stimulated	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
IL-1R	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
was	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
surface	NN	O	B-protein
IL-4R	NN	O	I-protein
because	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
these	NN	O	O
cytokines	NN	O	O
did	NN	O	O
not	NN	O	O
decrease	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
binding	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
suppression	NN	O	O
of	NN	O	O
IL-1R	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
was	NN	O	O
associated	NN	O	O
with	NN	O	O
decreased	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4/IL-13-inducible	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
Stat6	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
may	NN	O	O
mediate	NN	O	O
their	NN	O	O
suppressive	NN	O	O
effects	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
certain	NN	O	O
cytokines	NN	O	O
,	NN	O	O
including	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
antagonize	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1RI	NN	O	O
and	NN	O	O
IL-1RII	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Cytokine	NN	O	B-protein
rescue	NN	O	O
from	NN	O	O
glucocorticoid	NN	O	O
induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
DEX	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
synthetic	NN	O	O
glucocorticoid	NN	O	O
,	NN	O	O
causes	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_line
Th	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
induction	NN	O	O
of	NN	O	O
cell	NN	O	O
death	NN	O	O
is	NN	O	O
prevented	NN	O	O
by	NN	O	O
specific	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
namely	NN	O	O
,	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
in	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
differential	NN	O	O
rescue	NN	O	O
by	NN	O	O
specific	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
Th	NN	O	B-cell_type
cells	NN	O	I-cell_type
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
that	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
DEX	NN	O	O
and	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

In	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
the	NN	O	O
cellular	NN	O	O
levels	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
were	NN	O	O
evaluated	NN	O	O
by	NN	O	O
DEX	NN	O	O
treatment	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
DEX-mediated	NN	O	O
IkappaBalpha	NN	O	B-protein
induction	NN	O	O
was	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
the	NN	O	O
reverse	NN	O	O
profile	NN	O	O
was	NN	O	O
seen	NN	O	O
in	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
that	NN	O	O
inhibits	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
by	NN	O	O
DEX	NN	O	O
,	NN	O	O
also	NN	O	O
rescues	NN	O	O
these	NN	O	O
cells	NN	O	O
from	NN	O	O
DEX-induced	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
rescue	NN	O	O
cytokine	NN	O	O
is	NN	O	O
different	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_type
and	NN	O	I-cell_type
Th2	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
imply	NN	O	O
that	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
need	NN	O	O
to	NN	O	O
maintain	NN	O	O
a	NN	O	O
certain	NN	O	O
level	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
survive	NN	O	O
;	NN	O	O
up-	NN	O	O
or	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
NF	NN	O	I-protein
kappaB	NN	O	I-protein
through	NN	O	O
modulation	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
or	NN	O	O
DEX	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
cell	NN	O	O
survival	NN	O	O
or	NN	O	O
cell	NN	O	O
death	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
transcription	NN	O	B-DNA
factor	NN	O	I-DNA
genes	NN	O	I-DNA
after	NN	O	O
influenza	NN	O	O
A	NN	O	O
virus	NN	O	O
infection	NN	O	O
.	NN	O	O

Infection	NN	O	O
of	NN	O	O
human	NN	O	O
monocytes	NN	O	B-cell_type
with	NN	O	O
influenza	NN	O	O
A	NN	O	O
virus	NN	O	O
induces	NN	O	O
a	NN	O	O
broad	NN	O	O
range	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
and	NN	O	O
mononuclear	NN	O	O
cell	NN	O	O
attracting	NN	O	O
chemokines	NN	O	B-protein
before	NN	O	O
the	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
undergo	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
underlying	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
the	NN	O	O
corresponding	NN	O	O
genes	NN	O	O
are	NN	O	O
transcriptionally	NN	O	O
initiated	NN	O	O
after	NN	O	O
virus	NN	O	O
infection	NN	O	O
are	NN	O	O
still	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
seems	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
many	NN	O	O
proinflammatory	NN	O	B-DNA
cytokine	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
the	NN	O	O
only	NN	O	O
mechanism	NN	O	O
,	NN	O	O
since	NN	O	O
several	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
lack	NN	O	O
respective	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
their	NN	O	O
promoter	NN	O	B-DNA
regions	NN	O	I-DNA
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
additionally	NN	O	O
investigated	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
possible	NN	O	O
importance	NN	O	O
such	NN	O	O
as	NN	O	O
CREB	NN	O	B-protein
,	NN	O	O
CTF	NN	O	B-protein
,	NN	O	O
OTF-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
OTF-2	NN	O	B-protein
.	NN	O	O

To	NN	O	O
explore	NN	O	O
long-term	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
on	NN	O	O
the	NN	O	O
gene	NN	O	O
expression	NN	O	O
level	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
to	NN	O	O
substitute	NN	O	O
for	NN	O	O
metabolized	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
proteins	NN	O	I-protein
after	NN	O	O
their	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
cell-type-specific	NN	O	O
differential	NN	O	O
response	NN	O	O
:	NN	O	O
CREB	NN	O	B-DNA
,	NN	O	I-DNA
CTF	NN	O	I-DNA
,	NN	O	I-DNA
OTF-1	NN	O	I-DNA
,	NN	O	I-DNA
OFT-2	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
genes	NN	O	I-DNA
were	NN	O	O
strongly	NN	O	O
induced	NN	O	O
1	NN	O	O
to	NN	O	O
4	NN	O	O
hours	NN	O	O
after	NN	O	O
influenza	NN	O	O
A	NN	O	O
virus	NN	O	O
infection	NN	O	O
in	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
in	NN	O	O
freshly	NN	O	O
prepared	NN	O	O
human	NN	O	O
monocytes	NN	O	B-cell_type
no	NN	O	O
significant	NN	O	O
changes	NN	O	O
were	NN	O	O
detected	NN	O	O
.	NN	O	O

In	NN	O	O
infected	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
which	NN	O	O
die	NN	O	O
by	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
,	NN	O	O
CTF	NN	O	B-protein
,	NN	O	O
and	NN	O	O
OTF-2	NN	O	B-protein
was	NN	O	O
rather	NN	O	O
suppressed	NN	O	O
8	NN	O	O
hours	NN	O	O
after	NN	O	O
infection	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
the	NN	O	O
long-term	NN	O	O
regulation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
non-proliferating	NN	O	B-cell_type
cells	NN	O	I-cell_type
seems	NN	O	O
to	NN	O	O
be	NN	O	O
of	NN	O	O
minor	NN	O	O
importance	NN	O	O
after	NN	O	O
influenza	NN	O	O
infection	NN	O	O
since	NN	O	O
in	NN	O	O
apoptosisprone	NN	O	B-cell_type
cells	NN	O	I-cell_type
an	NN	O	O
immediate	NN	O	O
availability	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
proteins	NN	O	I-protein
is	NN	O	O
required	NN	O	O
.	NN	O	O

-DOCSTART-	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
diabetic	NN	O	O
nephropathy	NN	O	O
show	NN	O	O
increased	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
oxidative-stress	NN	O	B-protein
sensitive	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

Increased	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
and	NN	O	O
subsequent	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
NF-kappaB	NN	O	B-protein
has	NN	O	O
been	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
late	NN	O	O
diabetic	NN	O	O
complications	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
dependent	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
evident	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
diabetic	NN	O	O
nephropathy	NN	O	O
we	NN	O	O
used	NN	O	O
an	NN	O	O
Electrophoretic	NN	O	O
Mobility	NN	O	O
Shift	NN	O	O
Assay	NN	O	O
based	NN	O	O
semiquantitative	NN	O	O
detection	NN	O	O
system	NN	O	O
which	NN	O	O
enabled	NN	O	O
us	NN	O	O
to	NN	O	O
determine	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
ex	NN	O	O
vivo	NN	O	O
isolated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
examined	NN	O	O
33	NN	O	O
patients	NN	O	O
with	NN	O	O
diabetes	NN	O	O
mellitus	NN	O	O
(	NN	O	O
Type	NN	O	O
I	NN	O	O
and	NN	O	O
Type	NN	O	O
II	NN	O	O
)	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
diabetic	NN	O	O
nephropathy	NN	O	O
showed	NN	O	O
higher	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
Electrophoretic	NN	O	O
Mobility	NN	O	O
Shift	NN	O	O
Assays	NN	O	O
and	NN	O	O
stronger	NN	O	O
immunohistological	NN	O	O
staining	NN	O	O
for	NN	O	O
activated	NN	O	O
NF-kappaBp65	NN	O	B-protein
than	NN	O	O
patients	NN	O	O
without	NN	O	O
renal	NN	O	O
complications	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
albuminuria	NN	O	O
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.316	NN	O	O
)	NN	O	O
and	NN	O	O
with	NN	O	O
thrombomodulin	NN	O	B-protein
plasma	NN	O	O
concentrations	NN	O	O
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.33	NN	O	O
)	NN	O	O
,	NN	O	O
indicative	NN	O	O
for	NN	O	O
albuminuria	NN	O	O
associated	NN	O	O
endothelial	NN	O	O
dysfunction	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
3	NN	O	O
day	NN	O	O
intervention	NN	O	O
study	NN	O	O
in	NN	O	O
which	NN	O	O
600	NN	O	O
mg	NN	O	O
of	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
thioctic	NN	O	O
acid	NN	O	O
(	NN	O	O
alpha-lipoic	NN	O	O
acid	NN	O	O
)	NN	O	O
per	NN	O	O
day	NN	O	O
were	NN	O	O
given	NN	O	O
to	NN	O	O
nine	NN	O	O
patients	NN	O	O
with	NN	O	O
diabetic	NN	O	O
nephropathy	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
in	NN	O	O
plasma	NN	O	O
samples	NN	O	O
was	NN	O	O
decreased	NN	O	O
by	NN	O	O
48	NN	O	O
%	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
ex	NN	O	O
vivo	NN	O	O
isolated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
38	NN	O	O
%	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
in	NN	O	O
ex	NN	O	O
vivo	NN	O	O
isolated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
diabetes	NN	O	O
mellitus	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
diabetic	NN	O	O
nephropathy	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
dependent	NN	O	O
on	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
since	NN	O	O
thioctic	NN	O	O
acid	NN	O	O
(	NN	O	O
alpha-lipoic	NN	O	O
acid	NN	O	O
)	NN	O	O
reduced	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Osteoclast	NN	O	B-protein
markers	NN	O	I-protein
accumulate	NN	O	O
on	NN	O	O
cells	NN	O	O
developing	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
precursors	NN	O	I-cell_type
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
show	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
osteoclasts	NN	O	I-cell_type
develop	NN	O	O
in	NN	O	O
vitro	NN	O	O
from	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
however	NN	O	O
,	NN	O	O
special	NN	O	O
cultures	NN	O	O
conditions	NN	O	O
and/or	NN	O	O
cytokine	NN	O	O
mobilized	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
are	NN	O	O
apparently	NN	O	O
required	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
cells	NN	O	O
expressing	NN	O	O
osteoclast	NN	O	B-protein
markers	NN	O	I-protein
differentiate	NN	O	O
from	NN	O	O
precursors	NN	O	O
present	NN	O	O
in	NN	O	O
nonmobilized	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
without	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
stromal	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
cytokines	NN	O	O
or	NN	O	O
steroids	NN	O	O
;	NN	O	O
and	NN	O	O
characterize	NN	O	O
their	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Three	NN	O	O
days	NN	O	O
after	NN	O	O
establishing	NN	O	O
high-density	NN	O	O
PBMC	NN	O	B-cell_type
cultures	NN	O	O
(	NN	O	O
1.5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells/cm2	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
serum-containing	NN	O	O
medium	NN	O	O
,	NN	O	O
small	NN	O	O
adherent	NN	O	B-cell_line
colonies	NN	O	I-cell_line
of	NN	O	O
tartrate	NN	O	B-cell_line
resistant	NN	O	I-cell_line
acid	NN	O	I-cell_line
phosphatase	NN	O	I-cell_line
positive	NN	O	I-cell_line
(	NN	O	I-cell_line
TRAP+	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
emerge	NN	O	O
,	NN	O	O
amidst	NN	O	O
massive	NN	O	O
monocyte	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

These	NN	O	O
adherent	NN	O	B-cell_type
cells	NN	O	I-cell_type
have	NN	O	O
an	NN	O	O
eccentrically	NN	O	O
placed	NN	O	O
,	NN	O	O
round	NN	O	O
nucleus	NN	O	O
,	NN	O	O
and	NN	O	O
express	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
TRAP	NN	O	B-protein
and	NN	O	O
sodium	NN	O	B-protein
fluoride-resistant-	NN	O	I-protein
alpha-naphthyl-acetate-esterase	NN	O	I-protein
(	NN	O	O
NaF-R-NSE	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Over	NN	O	O
the	NN	O	O
next	NN	O	O
week	NN	O	O
,	NN	O	O
this	NN	O	O
cell	NN	O	O
population	NN	O	O
accumulates	NN	O	O
phenotypic	NN	O	O
markers	NN	O	O
of	NN	O	O
osteoclasts	NN	O	B-cell_type
(	NN	O	O
vitronectin	NN	O	B-protein
receptor	NN	O	I-protein
[	NN	O	O
VR	NN	O	B-protein
]	NN	O	O
,	NN	O	O
calcitonin	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
TRAP	NN	O	B-protein
,	NN	O	O
cathepsin	NN	O	B-protein
K	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
mRNA	NN	O	B-RNA
)	NN	O	O
with	NN	O	O
increased	NN	O	O
nuclearity	NN	O	O
,	NN	O	O
covering	NN	O	O
the	NN	O	O
entire	NN	O	O
surface	NN	O	O
by	NN	O	O
15	NN	O	O
days	NN	O	O
.	NN	O	O

When	NN	O	O
cultured	NN	O	O
on	NN	O	O
bone	NN	O	O
,	NN	O	O
VR+	NN	O	B-cell_line
,	NN	O	I-cell_line
TRAP+	NN	O	I-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
low	NN	O	O
multinuclearity	NN	O	O
appear	NN	O	O
and	NN	O	O
cover	NN	O	O
up	NN	O	O
to	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
surface	NN	O	O
.	NN	O	O

Resorption	NN	O	O
lacunae	NN	O	O
can	NN	O	O
be	NN	O	O
observed	NN	O	O
by	NN	O	O
day	NN	O	O
22	NN	O	O
.	NN	O	O

Although	NN	O	O
these	NN	O	O
pits	NN	O	O
are	NN	O	O
not	NN	O	O
nearly	NN	O	O
as	NN	O	O
numerous	NN	O	O
as	NN	O	O
the	NN	O	O
cells	NN	O	O
of	NN	O	O
preosteoclast	NN	O	B-cell_type
phenotype	NN	O	I-cell_type
,	NN	O	O
they	NN	O	O
do	NN	O	O
represent	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
osteoclast-like	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
has	NN	O	O
achieved	NN	O	O
osteoclastic	NN	O	O
maturity	NN	O	O
under	NN	O	O
these	NN	O	O
culture	NN	O	O
conditions	NN	O	O
.	NN	O	O

Transcripts	NN	O	O
for	NN	O	O
osteoprotegerin	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
OPGL	NN	O	B-protein
)	NN	O	O
,	NN	O	O
an	NN	O	O
osteoclast	NN	O	B-protein
differentiation	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
RANKL	NN	O	B-protein
and	NN	O	O
TRANCE	NN	O	B-protein
)	NN	O	O
are	NN	O	O
expressed	NN	O	O
,	NN	O	O
likely	NN	O	O
by	NN	O	O
adherent	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
an	NN	O	O
adherent	NN	O	O
population	NN	O	O
of	NN	O	O
cells	NN	O	O
,	NN	O	O
with	NN	O	O
preosteoclast/	NN	O	O
osteoclast	NN	O	B-cell_type
phenotypic	NN	O	O
properties	NN	O	O
,	NN	O	O
arises	NN	O	O
selectively	NN	O	O
under	NN	O	O
simple	NN	O	O
culture	NN	O	O
conditions	NN	O	O
from	NN	O	O
normal	NN	O	O
PBMC	NN	O	B-cell_type
.	NN	O	O

Further	NN	O	O
characterization	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
should	NN	O	O
identify	NN	O	O
factors	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
growth	NN	O	O
,	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
osteoclasts	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Oleic	NN	O	O
acid	NN	O	O
inhibits	NN	O	O
endothelial	NN	O	O
activation	NN	O	O
:	NN	O	O
A	NN	O	O
direct	NN	O	O
vascular	NN	O	O
antiatherogenic	NN	O	O
mechanism	NN	O	O
of	NN	O	O
a	NN	O	O
nutritional	NN	O	O
component	NN	O	O
in	NN	O	O
the	NN	O	O
mediterranean	NN	O	O
diet	NN	O	O
.	NN	O	O

Because	NN	O	O
oleic	NN	O	O
acid	NN	O	O
is	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
antiatherogenic	NN	O	O
effects	NN	O	O
attributed	NN	O	O
to	NN	O	O
the	NN	O	O
Mediterranean	NN	O	O
diet	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
this	NN	O	O
fatty	NN	O	O
acid	NN	O	O
can	NN	O	O
modulate	NN	O	O
endothelial	NN	O	O
activation	NN	O	O
,	NN	O	O
ie	NN	O	O
,	NN	O	O
the	NN	O	O
concerted	NN	O	O
expression	NN	O	O
of	NN	O	O
gene	NN	O	B-protein
products	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
leukocyte	NN	O	O
recruitment	NN	O	O
and	NN	O	O
early	NN	O	O
atherogenesis	NN	O	O
.	NN	O	O

We	NN	O	O
incubated	NN	O	O
sodium	NN	O	O
oleate	NN	O	O
with	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
0	NN	O	O
to	NN	O	O
72	NN	O	O
hours	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
coincubation	NN	O	O
of	NN	O	O
oleate	NN	O	O
with	NN	O	O
human	NN	O	B-protein
recombinant	NN	O	I-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-1alpha	NN	O	I-protein
,	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
for	NN	O	O
a	NN	O	O
further	NN	O	O
6	NN	O	O
to	NN	O	O
24	NN	O	O
hours	NN	O	O
.	NN	O	O

The	NN	O	O
endothelial	NN	O	O
expression	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
was	NN	O	O
monitored	NN	O	O
by	NN	O	O
cell	NN	O	O
surface	NN	O	O
enzyme	NN	O	O
immunoassays	NN	O	O
or	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
,	NN	O	O
and	NN	O	O
steady-state	NN	O	O
levels	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
were	NN	O	O
assessed	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

At	NN	O	O
10	NN	O	O
to	NN	O	O
100	NN	O	O
micromol/L	NN	O	O
for	NN	O	O
>	NN	O	O
24	NN	O	O
hours	NN	O	O
,	NN	O	O
oleate	NN	O	O
inhibited	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
all	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
tested	NN	O	O
.	NN	O	O

After	NN	O	O
a	NN	O	O
72-hour	NN	O	O
incubation	NN	O	O
with	NN	O	O
oleate	NN	O	O
and	NN	O	O
a	NN	O	O
further	NN	O	O
16-hour	NN	O	O
incubation	NN	O	O
with	NN	O	O
oleate	NN	O	O
plus	NN	O	O
1	NN	O	O
microg/mL	NN	O	O
LPS	NN	O	O
,	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
reduced	NN	O	O
by	NN	O	O
>	NN	O	O
40	NN	O	O
%	NN	O	O
compared	NN	O	O
with	NN	O	O
control	NN	O	O
.	NN	O	O

Adhesion	NN	O	O
of	NN	O	O
monocytoid	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
LPS-treated	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
was	NN	O	O
reduced	NN	O	O
concomitantly	NN	O	O
.	NN	O	O

Oleate	NN	O	O
also	NN	O	O
produced	NN	O	O
a	NN	O	O
quantitatively	NN	O	O
similar	NN	O	O
reduction	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
on	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
and	NN	O	O
inhibited	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
activation	NN	O	O
on	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
oleate	NN	O	O
for	NN	O	O
72	NN	O	O
hours	NN	O	O
decreased	NN	O	O
the	NN	O	O
relative	NN	O	O
proportions	NN	O	O
of	NN	O	O
saturated	NN	O	O
(	NN	O	O
palmitic	NN	O	O
and	NN	O	O
stearic	NN	O	O
)	NN	O	O
acids	NN	O	O
in	NN	O	O
total	NN	O	O
cell	NN	O	O
lipids	NN	O	O
and	NN	O	O
increased	NN	O	O
the	NN	O	O
proportions	NN	O	O
of	NN	O	O
oleate	NN	O	O
in	NN	O	O
total	NN	O	O
cell	NN	O	O
lipids	NN	O	O
without	NN	O	O
significantly	NN	O	O
changing	NN	O	O
the	NN	O	O
relative	NN	O	O
proportions	NN	O	O
of	NN	O	O
polyunsaturated	NN	O	O
fatty	NN	O	O
acids	NN	O	O
.	NN	O	O

Although	NN	O	O
less	NN	O	O
potent	NN	O	O
than	NN	O	O
polyunsaturated	NN	O	O
fatty	NN	O	O
acids	NN	O	O
in	NN	O	O
inhibiting	NN	O	O
endothelial	NN	O	O
activation	NN	O	O
,	NN	O	O
oleic	NN	O	O
acid	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
prevention	NN	O	O
of	NN	O	O
atherogenesis	NN	O	O
through	NN	O	O
selective	NN	O	O
displacement	NN	O	O
of	NN	O	O
saturated	NN	O	O
fatty	NN	O	O
acids	NN	O	O
in	NN	O	O
cell	NN	O	O
membrane	NN	O	O
phospholipids	NN	O	O
and	NN	O	O
a	NN	O	O
consequent	NN	O	O
modulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
for	NN	O	O
molecules	NN	O	O
involved	NN	O	O
in	NN	O	O
monocyte	NN	O	O
recruitment	NN	O	O
.	NN	O	O

-DOCSTART-	O

Both	NN	O	O
Stat3	NN	O	B-protein
-activation	NN	O	O
and	NN	O	O
Stat3	NN	O	B-protein
-independent	NN	O	O
BCL2	NN	O	B-protein
downregulation	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
interleukin-6	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
1A9-M	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
unique	NN	O	O
subclone	NN	O	O
of	NN	O	O
a	NN	O	O
bone	NN	O	B-cell_line
marrow-derived	NN	O	I-cell_line
stromal	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
BMS2.4	NN	O	B-cell_line
,	NN	O	O
produces	NN	O	O
soluble	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
inhibit	NN	O	O
proliferation	NN	O	O
of	NN	O	O
several	NN	O	O
types	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

An	NN	O	O
understanding	NN	O	O
of	NN	O	O
these	NN	O	O
molecules	NN	O	O
may	NN	O	O
be	NN	O	O
informative	NN	O	O
about	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
circuits	NN	O	O
that	NN	O	O
can	NN	O	O
potentially	NN	O	O
limit	NN	O	O
blood	NN	O	O
cell	NN	O	O
formation	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
expression	NN	O	O
cloning	NN	O	O
to	NN	O	O
identify	NN	O	O
interleukin-6	NN	O	B-protein
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
as	NN	O	O
one	NN	O	O
factor	NN	O	O
that	NN	O	O
suppressed	NN	O	O
growth	NN	O	O
of	NN	O	O
a	NN	O	O
pre-B-cell	NN	O	B-cell_line
variant	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
1A9-M	NN	O	B-cell_line
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
IL-6	NN	O	B-protein
induced	NN	O	O
macrophage-differentiation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
1A9-M	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

During	NN	O	O
this	NN	O	O
process	NN	O	O
,	NN	O	O
IL-6	NN	O	B-protein
downregulated	NN	O	O
expression	NN	O	O
of	NN	O	O
BCL2	NN	O	B-protein
in	NN	O	O
1A9-M	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
stimulated	NN	O	O
BCL-XL	NN	O	O
expression	NN	O	O
,	NN	O	O
but	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
p53	NN	O	O
,	NN	O	O
Bax	NN	O	O
,	NN	O	O
or	NN	O	O
Bak	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Mechanisms	NN	O	O
for	NN	O	O
transduction	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
-induced	NN	O	O
signals	NN	O	O
were	NN	O	O
then	NN	O	O
evaluated	NN	O	O
in	NN	O	O
IL-6-stimulated	NN	O	B-cell_line
1A9-M	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Whereas	NN	O	O
the	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
Stat3	NN	O	B-protein
)	NN	O	O
was	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
activated	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
either	NN	O	O
Stat1	NN	O	B-protein
or	NN	O	O
Stat5	NN	O	B-protein
.	NN	O	O

The	NN	O	O
importance	NN	O	O
of	NN	O	O
BCL2	NN	O	B-protein
and	NN	O	O
Stat3	NN	O	B-protein
on	NN	O	O
IL-6	NN	O	B-protein
-induced	NN	O	O
macrophage-differentiation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
studied	NN	O	O
with	NN	O	O
1A9-M	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
human	NN	O	B-protein
BCL2	NN	O	I-protein
or	NN	O	O
a	NN	O	O
dominant-negative	NN	O	O
form	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

IL-6	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
macrophage-differentiation	NN	O	O
,	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
continuously	NN	O	O
expressed	NN	O	O
BCL2	NN	O	B-protein
.	NN	O	O

A	NN	O	O
dominant-negative	NN	O	B-protein
form	NN	O	I-protein
of	NN	O	I-protein
Stat3	NN	O	I-protein
inhibited	NN	O	O
both	NN	O	O
macrophage-differentiation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-6	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
diminished	NN	O	O
Stat3	NN	O	B-protein
activity	NN	O	O
did	NN	O	O
not	NN	O	O
prevent	NN	O	O
IL-6	NN	O	B-protein
-induced	NN	O	O
downregulation	NN	O	O
of	NN	O	O
the	NN	O	O
BCL2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
IL-6	NN	O	B-protein
-induced	NN	O	O
macrophage-differentiation	NN	O	O
and	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
this	NN	O	O
model	NN	O	O
.	NN	O	O

Whereas	NN	O	O
overexpression	NN	O	O
of	NN	O	O
BCL2	NN	O	B-protein
abrogates	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
response	NN	O	O
,	NN	O	O
Stat3	NN	O	B-protein
-independent	NN	O	O
signals	NN	O	O
appear	NN	O	O
to	NN	O	O
downregulate	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
BCL2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
pathways	NN	O	O
in	NN	O	O
interleukin-8	NN	O	B-protein
production	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
polymorphonuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
lipopolysaccharide	NN	O	O
or	NN	O	O
Mycoplasma	NN	O	B-protein
fermentans	NN	O	I-protein
membrane	NN	O	I-protein
lipoproteins	NN	O	I-protein
.	NN	O	O

Interleukin-8	NN	O	B-protein
(	NN	O	O
IL-8	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
chemokine	NN	O	B-protein
that	NN	O	O
belongs	NN	O	O
to	NN	O	O
the	NN	O	O
alpha-chemokine	NN	O	B-protein
or	NN	O	O
CXC	NN	O	B-protein
subfamily	NN	O	I-protein
and	NN	O	O
is	NN	O	O
produced	NN	O	O
by	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
human	NN	O	O
cells	NN	O	O
,	NN	O	O
including	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
polymorphonuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PMN	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

IL-8	NN	O	B-protein
is	NN	O	O
secreted	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
inflammatory	NN	O	O
stimuli	NN	O	O
,	NN	O	O
notably	NN	O	O
bacterial	NN	O	O
products	NN	O	O
such	NN	O	O
as	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
these	NN	O	O
agents	NN	O	O
mediate	NN	O	O
IL-8	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
Mycoplasma	NN	O	B-protein
fermentans	NN	O	I-protein
lipid-associated	NN	O	I-protein
membrane	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
LAMPf	NN	O	B-protein
)	NN	O	O
induce	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
by	NN	O	O
THP-1	NN	O	B-cell_line
(	NN	O	I-cell_line
human	NN	O	I-cell_line
monocyte	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
PMN	NN	O	B-cell_type
at	NN	O	O
the	NN	O	O
same	NN	O	O
extent	NN	O	O
as	NN	O	O
LPS	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
monocytic	NN	O	O
cells	NN	O	O
with	NN	O	O
either	NN	O	O
LPS	NN	O	O
or	NN	O	O
LAMPf	NN	O	B-protein
led	NN	O	O
to	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
common	NN	O	O
downstream	NN	O	O
signaling	NN	O	O
events	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
of	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
cascades	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
PD-98059	NN	O	O
and	NN	O	O
SB203580	NN	O	O
,	NN	O	O
two	NN	O	O
potent	NN	O	O
and	NN	O	O
selective	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
MEK1	NN	O	B-protein
(	NN	O	O
a	NN	O	O
kinase	NN	O	B-protein
upstream	NN	O	O
of	NN	O	O
ERK1/2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
p38	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
both	NN	O	O
ERK1/2	NN	O	O
and	NN	O	O
p38	NN	O	O
cascades	NN	O	O
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
by	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
PMN	NN	O	B-cell_type
stimulated	NN	O	O
with	NN	O	O
bacterial	NN	O	O
fractions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
C3a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
(	NN	O	I-protein
desArg	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

The	NN	O	O
anaphylatoxin	NN	O	B-protein
C3a	NN	O	B-protein
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
have	NN	O	O
immunomodulatory	NN	O	O
effects	NN	O	O
on	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
C3a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
(	NN	O	I-protein
desArg	NN	O	I-protein
)	NN	O	I-protein
on	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
protein	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
in	NN	O	O
human	NN	O	O
PBMCs	NN	O	B-cell_type
,	NN	O	O
either	NN	O	O
alone	NN	O	O
or	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
LPS	NN	O	O
or	NN	O	O
IL-1beta	NN	O	B-protein
.	NN	O	O

C3a	NN	O	B-protein
or	NN	O	O
C3a	NN	O	B-protein
(	NN	O	I-protein
desArg	NN	O	I-protein
)	NN	O	I-protein
alone	NN	O	O
exhibited	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
or	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
when	NN	O	O
PBMC	NN	O	B-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
LPS	NN	O	O
or	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
both	NN	O	O
C3a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
(	NN	O	I-protein
desArg	NN	O	I-protein
)	NN	O	I-protein
were	NN	O	O
found	NN	O	O
to	NN	O	O
enhance	NN	O	O
IL-6	NN	O	B-protein
release	NN	O	O
by	NN	O	O
PBMC	NN	O	B-cell_type
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Since	NN	O	O
C3a	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
induce	NN	O	O
PGE2	NN	O	O
production	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
PGE2	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
influence	NN	O	O
cytokine	NN	O	O
production	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
PGE2	NN	O	O
in	NN	O	O
C3a	NN	O	B-protein
-mediated	NN	O	O
enhancement	NN	O	O
of	NN	O	O
LPS-	NN	O	O
and	NN	O	O
IL-1beta-induced	NN	O	O
IL-6	NN	O	O
production	NN	O	O
.	NN	O	O

Indomethacin	NN	O	O
blocked	NN	O	O
PGE2	NN	O	O
release	NN	O	O
,	NN	O	O
but	NN	O	O
had	NN	O	O
no	NN	O	O
influence	NN	O	O
on	NN	O	O
the	NN	O	O
observed	NN	O	O
effects	NN	O	O
of	NN	O	O
C3a	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
C3a	NN	O	B-protein
on	NN	O	O
IL-6	NN	O	B-protein
production	NN	O	O
are	NN	O	O
independent	NN	O	O
of	NN	O	O
PGE2	NN	O	O
formation	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
C3a	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
C3a	NN	O	B-protein
(	NN	O	I-protein
desArg	NN	O	I-protein
)	NN	O	I-protein
enhanced	NN	O	O
LPS	NN	O	O
-induced	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
for	NN	O	O
IL-6	NN	O	B-protein
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
PBMCs	NN	O	B-cell_type
with	NN	O	O
pertussis	NN	O	B-protein
toxin	NN	O	I-protein
blocked	NN	O	O
the	NN	O	O
functions	NN	O	O
of	NN	O	O
C3a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
(	NN	O	I-protein
desArg	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
molecules	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
G	NN	O	B-protein
protein	NN	O	I-protein
-coupled	NN	O	O
pathway	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
C3a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
(	NN	O	I-protein
desArg	NN	O	I-protein
)	NN	O	I-protein
on	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
activating	NN	O	B-protein
protein-1	NN	O	I-protein
binding	NN	O	O
.	NN	O	O

Both	NN	O	O
molecules	NN	O	O
enhanced	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
activating	NN	O	B-protein
protein-1	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
capacity	NN	O	O
of	NN	O	O
intact	NN	O	O
C3a	NN	O	B-protein
and	NN	O	O
its	NN	O	O
circulating	NN	O	O
des-Arg	NN	O	B-protein
form	NN	O	I-protein
to	NN	O	O
exert	NN	O	O
immunmodulatory	NN	O	O
effects	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

-DOCSTART-	O

Phenylarsine	NN	O	O
oxide	NN	O	O
inhibits	NN	O	O
ex	NN	O	O
vivo	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
.	NN	O	O

Phenylarsine	NN	O	O
oxide	NN	O	O
(	NN	O	O
PAO	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
described	NN	O	O
as	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
tyrosine	NN	O	B-protein
phosphatase	NN	O	I-protein
activity	NN	O	O
,	NN	O	O
inhibits	NN	O	O
H2O2	NN	O	O
release	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
electrochemistry	NN	O	O
.	NN	O	O

Since	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
replication	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
favored	NN	O	O
under	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
conditions	NN	O	O
,	NN	O	O
ex	NN	O	O
vivo	NN	O	O
experiments	NN	O	O
using	NN	O	O
uninfected	NN	O	O
PBMCs	NN	O	B-cell_type
,	NN	O	O
primary	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
or	NN	O	O
a	NN	O	O
latently	NN	O	O
infected	NN	O	O
promonocytic	NN	O	B-cell_line
U1	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
show	NN	O	O
that	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
and	NN	O	O
reactivation	NN	O	O
,	NN	O	O
monitored	NN	O	O
by	NN	O	O
p24	NN	O	B-protein
antigen	NN	O	I-protein
measurement	NN	O	O
,	NN	O	O
are	NN	O	O
inhibited	NN	O	O
by	NN	O	O
PAO	NN	O	O
in	NN	O	O
a	NN	O	O
time-	NN	O	O
and	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
can	NN	O	O
be	NN	O	O
linked	NN	O	O
with	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
when	NN	O	O
uninfected	NN	O	O
monocytes	NN	O	O
are	NN	O	O
induced	NN	O	O
by	NN	O	O
either	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
or	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Pharmacological	NN	O	O
control	NN	O	O
of	NN	O	O
antigen	NN	O	O
responsiveness	NN	O	O
in	NN	O	O
genetically	NN	O	B-cell_line
modified	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
chimeric	NN	O	B-DNA
TCR	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
comprising	NN	O	O
an	NN	O	O
anti-hapten	NN	O	B-protein
single-chain	NN	O	I-protein
Ab	NN	O	I-protein
variable	NN	O	I-protein
fragment	NN	O	I-protein
fused	NN	O	O
to	NN	O	O
the	NN	O	O
transmembrane	NN	O	B-protein
and	NN	O	I-protein
cytoplasmic	NN	O	I-protein
regions	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
TCR	NN	O	I-protein
zeta-chain	NN	O	I-protein
,	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
tetracycline-regulatable	NN	O	O
system	NN	O	O
could	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
regulate	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
single	NN	O	O
vector	NN	O	O
encoding	NN	O	O
the	NN	O	O
tetracycline	NN	O	B-protein
trans-activator	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
constitutive	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
TCR	NN	O	I-protein
,	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
a	NN	O	O
trans-activator	NN	O	B-DNA
protein-responsive	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
tetracyclines	NN	O	O
,	NN	O	O
the	NN	O	O
transfected	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
shown	NN	O	O
to	NN	O	O
express	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
receptor	NN	O	I-protein
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
and	NN	O	O
could	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
its	NN	O	O
cognate	NN	O	B-protein
Ag	NN	O	I-protein
,	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

When	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
increasing	NN	O	O
concentrations	NN	O	O
of	NN	O	O
tetracyclines	NN	O	O
,	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
receptor	NN	O	I-protein
was	NN	O	O
suppressed	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
suppression	NN	O	O
was	NN	O	O
sufficient	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
complete	NN	O	O
loss	NN	O	O
of	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
the	NN	O	O
targeted	NN	O	B-protein
Ag	NN	O	I-protein
.	NN	O	O

Prolonged	NN	O	O
suppression	NN	O	O
of	NN	O	O
trans-gene	NN	O	O
expression	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
7	NN	O	O
days	NN	O	O
was	NN	O	O
observed	NN	O	O
after	NN	O	O
doxycycline	NN	O	O
was	NN	O	O
removed	NN	O	O
from	NN	O	O
the	NN	O	O
cultures	NN	O	O
,	NN	O	O
but	NN	O	O
eventual	NN	O	O
recovery	NN	O	O
of	NN	O	O
surface	NN	O	O
expression	NN	O	O
was	NN	O	O
complete	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
absolute	NN	O	O
time	NN	O	O
to	NN	O	O
recovery	NN	O	O
was	NN	O	O
directly	NN	O	O
proportional	NN	O	O
to	NN	O	O
the	NN	O	O
initial	NN	O	O
concentration	NN	O	O
of	NN	O	O
the	NN	O	O
drug	NN	O	O
.	NN	O	O

Pharmacologic	NN	O	O
control	NN	O	O
of	NN	O	O
trans-gene	NN	O	O
expression	NN	O	O
in	NN	O	O
gene-modified	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
will	NN	O	O
not	NN	O	O
only	NN	O	O
facilitate	NN	O	O
new	NN	O	O
approaches	NN	O	O
to	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
different	NN	O	O
aspects	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
biology	NN	O	O
,	NN	O	O
but	NN	O	O
will	NN	O	O
also	NN	O	O
provide	NN	O	O
the	NN	O	O
basis	NN	O	O
for	NN	O	O
new	NN	O	O
gene	NN	O	O
therapy	NN	O	O
strategies	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
fibromyalgia	NN	O	O
and	NN	O	O
low	NN	O	O
back	NN	O	O
pain	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
fibromyalgia	NN	O	O
(	NN	O	O
FMS	NN	O	O
)	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
disorder	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
altered	NN	O	O
functioning	NN	O	O
of	NN	O	O
the	NN	O	O
stress	NN	O	O
response	NN	O	O
system	NN	O	O
.	NN	O	O

FMS	NN	O	O
patients	NN	O	O
display	NN	O	O
a	NN	O	O
hyperreactive	NN	O	O
pituitary	NN	O	O
adrenocorticotropic	NN	O	O
hormone	NN	O	O
(	NN	O	O
ACTH	NN	O	O
)	NN	O	O
release	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
corticotropin-releasing	NN	O	O
hormone	NN	O	O
(	NN	O	O
CRH	NN	O	O
)	NN	O	O
and	NN	O	O
to	NN	O	O
insulin-induced	NN	O	O
hypoglycemia	NN	O	O
.	NN	O	O

We	NN	O	O
suggested	NN	O	O
that	NN	O	O
negative	NN	O	O
feedback	NN	O	O
of	NN	O	O
cortisol	NN	O	O
could	NN	O	O
be	NN	O	O
deranged	NN	O	O
.	NN	O	O

Therefore	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
properties	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
FMS	NN	O	O
patients	NN	O	O
and	NN	O	O
compared	NN	O	O
the	NN	O	O
results	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
healthy	NN	O	O
persons	NN	O	O
and	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
low	NN	O	O
back	NN	O	O
pain	NN	O	O
(	NN	O	O
LBP	NN	O	O
a	NN	O	O
localized	NN	O	O
pain	NN	O	O
condition	NN	O	O
)	NN	O	O
.	NN	O	O

Forty	NN	O	O
primary	NN	O	O
FMS	NN	O	O
patients	NN	O	O
(	NN	O	O
F	NN	O	O
:	NN	O	O
M	NN	O	O
=	NN	O	O
36	NN	O	O
:	NN	O	O
4	NN	O	O
)	NN	O	O
,	NN	O	O
28	NN	O	O
LBP	NN	O	O
patients	NN	O	O
(	NN	O	O
25	NN	O	O
:	NN	O	O
3	NN	O	O
)	NN	O	O
and	NN	O	O
14	NN	O	O
(	NN	O	O
12	NN	O	O
:	NN	O	O
2	NN	O	O
)	NN	O	O
healthy	NN	O	O
,	NN	O	O
sedentary	NN	O	O
control	NN	O	O
persons	NN	O	O
were	NN	O	O
recruited	NN	O	O
for	NN	O	O
the	NN	O	O
study	NN	O	O
.	NN	O	O

Urinary	NN	O	O
free	NN	O	O
cortisol	NN	O	O
excretion	NN	O	O
in	NN	O	O
FMS	NN	O	O
and	NN	O	O
LBP	NN	O	O
patients	NN	O	O
was	NN	O	O
lower	NN	O	O
compared	NN	O	O
to	NN	O	O
controls	NN	O	O
.	NN	O	O

Only	NN	O	O
FMS	NN	O	O
patients	NN	O	O
displayed	NN	O	O
lower	NN	O	O
CBG	NN	O	O
and	NN	O	O
basal	NN	O	O
serum	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
when	NN	O	O
compared	NN	O	O
to	NN	O	O
controls	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
plasma	NN	O	O
free	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
were	NN	O	O
similar	NN	O	O
in	NN	O	O
the	NN	O	O
three	NN	O	O
groups	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
per	NN	O	O
cell	NN	O	O
among	NN	O	O
the	NN	O	O
three	NN	O	O
groups	NN	O	O
(	NN	O	O
FMS	NN	O	O
:	NN	O	O
6498	NN	O	O
+/-	NN	O	O
252	NN	O	O
,	NN	O	O
LBP	NN	O	O
:	NN	O	O
6625	NN	O	O
+/-	NN	O	O
284	NN	O	O
,	NN	O	O
controls	NN	O	O
:	NN	O	O
6576	NN	O	O
+/-	NN	O	O
304	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
FMS	NN	O	O
(	NN	O	O
14.5	NN	O	O
+/-	NN	O	O
0.9	NN	O	O
nmol/l	NN	O	O
)	NN	O	O
and	NN	O	O
LBP	NN	O	O
(	NN	O	O
14.7	NN	O	O
+/-	NN	O	O
1.3	NN	O	O
nmol/l	NN	O	O
)	NN	O	O
subjects	NN	O	O
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
controls	NN	O	O
(	NN	O	O
10.9	NN	O	O
+/-	NN	O	O
0.8	NN	O	O
nmol/l	NN	O	O
)	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
.05	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
maximal	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
the	NN	O	O
maximal	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
(	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
cortisol	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
FMS	NN	O	O
group	NN	O	O
was	NN	O	O
approximately	NN	O	O
1.5	NN	O	O
times	NN	O	O
higher	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
.05	NN	O	O
)	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
or	NN	O	O
LBP	NN	O	O
group	NN	O	O
.	NN	O	O

The	NN	O	O
ED50	NN	O	O
(	NN	O	O
the	NN	O	O
cortisol	NN	O	O
concentration	NN	O	O
giving	NN	O	O
50	NN	O	O
%	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
)	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
was	NN	O	O
identical	NN	O	O
in	NN	O	O
all	NN	O	O
three	NN	O	O
groups	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
FMS	NN	O	O
patients	NN	O	O
have	NN	O	O
a	NN	O	O
mild	NN	O	O
hypocortisolemia	NN	O	O
,	NN	O	O
increased	NN	O	O
cortisol	NN	O	O
feedback	NN	O	O
resistance	NN	O	O
in	NN	O	O
combination	NN	O	O
probably	NN	O	O
with	NN	O	O
a	NN	O	O
reduced	NN	O	O
CRH	NN	O	O
synthesis	NN	O	O
or	NN	O	O
release	NN	O	O
in	NN	O	O
the	NN	O	O
hypothalamus	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
and	NN	O	O
mineralocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
MR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
CRH	NN	O	O
regulation	NN	O	O
in	NN	O	O
the	NN	O	O
FMS	NN	O	O
patients	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
solved	NN	O	O
.	NN	O	O

-DOCSTART-	O

Temporal	NN	O	O
control	NN	O	O
of	NN	O	O
IgH	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
developing	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
the	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
locus	NN	O	I-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
suggested	NN	O	O
roles	NN	O	O
of	NN	O	O
the	NN	O	O
downstream	NN	O	B-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
regions	NN	O	I-DNA
acting	NN	O	O
as	NN	O	O
a	NN	O	O
Locus	NN	O	B-DNA
Control	NN	O	I-DNA
Region	NN	O	I-DNA
(	NN	O	O
LCR	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
have	NN	O	O
allowed	NN	O	O
comparisons	NN	O	O
to	NN	O	O
be	NN	O	O
made	NN	O	O
between	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
locus	NN	O	I-DNA
with	NN	O	O
other	NN	O	O
model	NN	O	O
systems	NN	O	O
whose	NN	O	O
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
governed	NN	O	O
by	NN	O	O
LCR	NN	O	B-DNA
activity	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
summarize	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
3'LCR	NN	O	I-DNA
and	NN	O	O
its	NN	O	O
putative	NN	O	O
functional	NN	O	O
role	NN	O	O
in	NN	O	O
IgH	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
and	NN	O	O
compare	NN	O	O
it	NN	O	O
with	NN	O	O
the	NN	O	O
5'LCR	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
locus	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

HLA	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
and	NN	O	O
generation	NN	O	O
of	NN	O	O
cytotoxic	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
against	NN	O	O
peptides	NN	O	O
derived	NN	O	O
from	NN	O	O
oncogenic	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

AIMS	NN	O	O
AND	NN	O	O
BACKGROUND	NN	O	O
:	NN	O	O
Structurally	NN	O	O
altered	NN	O	O
proteins	NN	O	O
(	NN	O	O
derived	NN	O	O
from	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
or	NN	O	O
gene	NN	O	O
mutations	NN	O	O
)	NN	O	O
can	NN	O	O
be	NN	O	O
considered	NN	O	O
tumor	NN	O	O
specific	NN	O	O
antigens	NN	O	O
and	NN	O	O
represent	NN	O	O
an	NN	O	O
attractive	NN	O	O
target	NN	O	O
for	NN	O	O
a	NN	O	O
T-cell	NN	O	O
mediated	NN	O	O
response	NN	O	O
.	NN	O	O

T	NN	O	O
lymphocytes	NN	O	O
recognize	NN	O	O
antigens	NN	O	O
in	NN	O	O
the	NN	O	O
form	NN	O	O
of	NN	O	O
peptides	NN	O	O
bound	NN	O	O
to	NN	O	O
HLA-molecules	NN	O	B-protein
.	NN	O	O

MATERIALS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
Peptides	NN	O	O
derived	NN	O	O
from	NN	O	O
oncogenic	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
screened	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
HLA	NN	O	B-protein
binding	NN	O	I-protein
motifs	NN	O	I-protein
;	NN	O	O
actual	NN	O	O
binding	NN	O	O
were	NN	O	O
evaluated	NN	O	O
by	NN	O	O
HLA	NN	O	O
stabilization	NN	O	O
experiments	NN	O	O
using	NN	O	O
transfectants	NN	O	O
and	NN	O	O
specific	NN	O	O
anti-HLA	NN	O	B-protein
antibodies	NN	O	I-protein
.	NN	O	O

Specific	NN	O	O
lymphocytes	NN	O	B-cell_type
were	NN	O	O
induced	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
peptide	NN	O	O
sensitization	NN	O	O
and	NN	O	O
screened	NN	O	O
by	NN	O	O
thymidine	NN	O	O
uptake	NN	O	O
or	NN	O	O
cellular	NN	O	O
cytotoxic	NN	O	O
assays	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
We	NN	O	O
identified	NN	O	O
peptides	NN	O	O
derived	NN	O	O
from	NN	O	O
EWS/FLI-1	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
from	NN	O	O
mutated	NN	O	B-protein
K-RAS	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
encompassing	NN	O	O
respectively	NN	O	O
the	NN	O	O
fusion	NN	O	B-protein
point	NN	O	I-protein
and	NN	O	O
the	NN	O	O
mutation	NN	O	O
at	NN	O	O
position	NN	O	B-protein
12	NN	O	I-protein
)	NN	O	O
that	NN	O	O
showed	NN	O	O
binding	NN	O	B-protein
motif	NN	O	I-protein
for	NN	O	O
HLA-Cw*0702	NN	O	B-protein
and	NN	O	O
HLA-A3	NN	O	B-protein
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
actual	NN	O	O
binding	NN	O	O
of	NN	O	O
these	NN	O	O
peptides	NN	O	O
was	NN	O	O
analysed	NN	O	O
in	NN	O	O
a	NN	O	O
stabilization	NN	O	O
assay	NN	O	O
.	NN	O	O

We	NN	O	O
detected	NN	O	O
binding	NN	O	O
for	NN	O	O
the	NN	O	O
EWS/FLI-1	NN	O	O
peptide	NN	O	O
and	NN	O	O
for	NN	O	O
5	NN	O	O
RAS	NN	O	O
peptides	NN	O	O
(	NN	O	O
1	NN	O	O
wild	NN	O	O
type	NN	O	O
and	NN	O	O
4	NN	O	O
mutated	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
temperature	NN	O	O
,	NN	O	O
beta	NN	O	B-protein
2-microglobulin	NN	O	I-protein
(	NN	O	O
beta	NN	O	B-protein
2-m	NN	O	I-protein
)	NN	O	O
and	NN	O	O
fetal	NN	O	O
calf	NN	O	O
serum	NN	O	O
(	NN	O	O
FCS	NN	O	O
)	NN	O	O
on	NN	O	O
the	NN	O	O
binding	NN	O	O
and	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
the	NN	O	O
HLA/peptide	NN	O	B-protein
complex	NN	O	I-protein
was	NN	O	O
studied	NN	O	O
.	NN	O	O

A	NN	O	O
low	NN	O	O
temperature	NN	O	O
(	NN	O	O
26	NN	O	O
degrees	NN	O	O
C	NN	O	O
)	NN	O	O
increased	NN	O	O
the	NN	O	O
binding	NN	O	O
both	NN	O	O
in	NN	O	O
HLA-A3	NN	O	B-protein
and	NN	O	O
HLA-Cw*0702	NN	O	B-protein
,	NN	O	O
while	NN	O	O
FCS	NN	O	O
reduced	NN	O	O
it	NN	O	O
.	NN	O	O

beta	NN	O	B-protein
2-m	NN	O	I-protein
increased	NN	O	O
the	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
HLA-A3	NN	O	B-protein
molecule	NN	O	I-protein
but	NN	O	O
did	NN	O	O
not	NN	O	O
influence	NN	O	O
the	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
HLA-Cw*0702	NN	O	B-protein
.	NN	O	O

The	NN	O	O
stability	NN	O	O
of	NN	O	O
already	NN	O	O
formed	NN	O	O
complexed	NN	O	O
was	NN	O	O
somewhat	NN	O	O
different	NN	O	O
in	NN	O	O
the	NN	O	O
HLA-A3	NN	O	B-protein
and	NN	O	O
HLA-Cw*0702	NN	O	B-protein
system	NN	O	O
:	NN	O	O
both	NN	O	O
were	NN	O	O
more	NN	O	O
stable	NN	O	O
at	NN	O	O
26	NN	O	O
degrees	NN	O	O
C	NN	O	O
than	NN	O	O
at	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
but	NN	O	O
while	NN	O	O
the	NN	O	O
beta	NN	O	B-protein
2-m	NN	O	I-protein
and	NN	O	O
FCS	NN	O	O
did	NN	O	O
not	NN	O	O
influence	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
the	NN	O	O
HLA-A3/peptide	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
they	NN	O	O
seemed	NN	O	O
to	NN	O	O
cause	NN	O	O
opposite	NN	O	O
effects	NN	O	O
in	NN	O	O
the	NN	O	O
HLA-Cw*0702	NN	O	B-protein
system	NN	O	O
(	NN	O	O
beta	NN	O	B-protein
2-m	NN	O	I-protein
stabilized	NN	O	O
and	NN	O	O
FCS	NN	O	O
destabilized	NN	O	O
the	NN	O	O
complex	NN	O	O
)	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
generate	NN	O	O
a	NN	O	O
specific	NN	O	O
CD8+	NN	O	B-cell_line
CTL	NN	O	I-cell_line
line	NN	O	I-cell_line
against	NN	O	O
a	NN	O	O
K-RAS	NN	O	O
mutated	NN	O	O
peptide	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Although	NN	O	O
binding	NN	O	B-protein
motifs	NN	O	I-protein
and	NN	O	O
actual	NN	O	O
HLA	NN	O	O
binding	NN	O	O
can	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
several	NN	O	O
cases	NN	O	O
,	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
a	NN	O	O
cellular	NN	O	O
response	NN	O	O
is	NN	O	O
infrequent	NN	O	O
,	NN	O	O
confirming	NN	O	O
that	NN	O	O
HLA	NN	O	O
binding	NN	O	O
is	NN	O	O
necessary	NN	O	O
but	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
obtain	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
response	NN	O	O
.	NN	O	O

Further	NN	O	O
optimization	NN	O	O
of	NN	O	O
culture	NN	O	O
conditions	NN	O	O
,	NN	O	O
type	NN	O	O
of	NN	O	O
Antigen	NN	O	B-cell_type
Presenting	NN	O	I-cell_type
Cells	NN	O	I-cell_type
(	NN	O	O
APC	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
peptides	NN	O	O
,	NN	O	O
use	NN	O	O
of	NN	O	O
stabilizers	NN	O	O
like	NN	O	O
beta	NN	O	B-protein
2-m	NN	O	I-protein
are	NN	O	O
still	NN	O	O
needed	NN	O	O
.	NN	O	O

-DOCSTART-	O

OCA-B	NN	O	B-protein
is	NN	O	O
a	NN	O	O
functional	NN	O	O
analog	NN	O	O
of	NN	O	O
VP16	NN	O	B-protein
but	NN	O	O
targets	NN	O	O
a	NN	O	O
separate	NN	O	O
surface	NN	O	O
of	NN	O	O
the	NN	O	O
Oct-1	NN	O	B-protein
POU	NN	O	I-protein
domain	NN	O	I-protein
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
Mol	NN	O	O
Cell	NN	O	O
Biol	NN	O	O
1998	NN	O	O
Apr	NN	O	O
;	NN	O	O
18	NN	O	O
(	NN	O	O
4	NN	O	O
)	NN	O	O
:	NN	O	O
2430	NN	O	O
]	NN	O	O

OCA-B	NN	O	B-protein
is	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	B-protein
coregulator	NN	O	I-protein
of	NN	O	O
the	NN	O	O
broadly	NN	O	O
expressed	NN	O	O
POU	NN	O	B-protein
domain	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Oct-1	NN	O	B-protein
.	NN	O	O

OCA-B	NN	O	B-protein
associates	NN	O	O
with	NN	O	O
the	NN	O	O
Oct-1	NN	O	B-protein
POU	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
a	NN	O	O
bipartite	NN	O	O
DNA-binding	NN	O	O
structure	NN	O	O
containing	NN	O	O
a	NN	O	O
POU-specific	NN	O	B-protein
(	NN	O	I-protein
POU	NN	O	I-protein
[	NN	O	I-protein
S	NN	O	I-protein
]	NN	O	I-protein
)	NN	O	I-protein
domain	NN	O	I-protein
joined	NN	O	O
by	NN	O	O
a	NN	O	O
flexible	NN	O	O
linker	NN	O	O
to	NN	O	O
a	NN	O	O
POU	NN	O	B-protein
homeodomain	NN	O	I-protein
(	NN	O	O
POU	NN	O	B-protein
[	NN	O	I-protein
H	NN	O	I-protein
]	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
OCA-B	NN	O	B-protein
alters	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
Oct-1	NN	O	B-protein
in	NN	O	O
two	NN	O	O
ways	NN	O	O
.	NN	O	O

It	NN	O	O
provides	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
domain	NN	O	O
which	NN	O	O
,	NN	O	O
unlike	NN	O	O
Oct-1	NN	O	B-protein
,	NN	O	O
activates	NN	O	O
an	NN	O	O
mRNA-type	NN	O	B-DNA
promoter	NN	O	I-DNA
effectively	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
stabilizes	NN	O	O
Oct-1	NN	O	B-protein
on	NN	O	O
the	NN	O	O
Oct-1-responsive	NN	O	B-DNA
octamer	NN	O	I-DNA
sequence	NN	O	I-DNA
ATGCAAAT	NN	O	O
.	NN	O	O

These	NN	O	O
properties	NN	O	O
of	NN	O	O
OCA-B	NN	O	B-protein
parallel	NN	O	O
those	NN	O	O
displayed	NN	O	O
by	NN	O	O
the	NN	O	O
herpes	NN	O	B-protein
simplex	NN	O	I-protein
virus	NN	O	I-protein
Oct-1	NN	O	I-protein
coregulator	NN	O	I-protein
VP16	NN	O	B-protein
.	NN	O	O

OCA-B	NN	O	B-protein
,	NN	O	O
however	NN	O	O
,	NN	O	O
interacts	NN	O	O
with	NN	O	O
a	NN	O	O
different	NN	O	O
surface	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-bound	NN	O	O
Oct-1	NN	O	B-protein
POU	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
interacting	NN	O	O
with	NN	O	O
both	NN	O	O
the	NN	O	O
POU	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
POU	NN	O	I-protein
(	NN	O	I-protein
H	NN	O	I-protein
)	NN	O	I-protein
domains	NN	O	I-protein
and	NN	O	O
the	NN	O	O
center	NN	O	O
of	NN	O	O
the	NN	O	O
ATGCAAAT	NN	O	B-DNA
octamer	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
OCA-B	NN	O	O
and	NN	O	O
VP16	NN	O	B-protein
interactions	NN	O	O
with	NN	O	O
the	NN	O	O
Oct-1	NN	O	B-protein
POU	NN	O	I-protein
domain	NN	O	I-protein
are	NN	O	O
sufficiently	NN	O	O
different	NN	O	O
to	NN	O	O
permit	NN	O	O
OCA-B	NN	O	B-protein
and	NN	O	O
VP16	NN	O	B-protein
to	NN	O	O
bind	NN	O	O
the	NN	O	O
Oct-1	NN	O	B-protein
POU	NN	O	I-protein
domain	NN	O	I-protein
simultaneously	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
emphasize	NN	O	O
the	NN	O	O
structural	NN	O	O
versatility	NN	O	O
of	NN	O	O
the	NN	O	O
Oct-1	NN	O	B-protein
POU	NN	O	I-protein
domain	NN	O	I-protein
in	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
coregulators	NN	O	O
.	NN	O	O

-DOCSTART-	O

Phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
couples	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
to	NN	O	O
the	NN	O	O
cell	NN	O	B-protein
cycle	NN	O	I-protein
regulator	NN	O	I-protein
E2F	NN	O	B-protein
.	NN	O	O

Cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
initiated	NN	O	O
by	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
critical	NN	O	O
for	NN	O	O
lymphoproliferation	NN	O	O
and	NN	O	O
an	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Phosphatidyl	NN	O	B-protein
inositol	NN	O	I-protein
3-kinase	NN	O	I-protein
(	NN	O	O
PI3K	NN	O	B-protein
)	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
nuclear	NN	O	O
targets	NN	O	O
for	NN	O	O
PI3K	NN	O	B-protein
are	NN	O	O
not	NN	O	O
known	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
identify	NN	O	O
the	NN	O	O
cell	NN	O	B-protein
cycle	NN	O	I-protein
regulator	NN	O	I-protein
E2F	NN	O	B-protein
as	NN	O	O
an	NN	O	O
IL-2	NN	O	B-protein
target	NN	O	O
in	NN	O	O
T	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
PI3K	NN	O	B-protein
as	NN	O	O
the	NN	O	O
critical	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
eliminate	NN	O	O
both	NN	O	O
Stat5	NN	O	B-protein
and	NN	O	O
Raf	NN	O	B-protein
/MEK	NN	O	B-protein
pathways	NN	O	O
from	NN	O	O
E2F	NN	O	B-protein
regulation	NN	O	O
.	NN	O	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
PKB	NN	O	B-protein
)	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
via	NN	O	O
PI3K	NN	O	B-protein
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
active	NN	O	O
PKB	NN	O	B-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
induce	NN	O	O
E2F	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
PI3K	NN	O	B-protein
inhibits	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Rb	NN	O	B-protein
,	NN	O	O
induction	NN	O	O
of	NN	O	O
cyclin	NN	O	B-protein
D3	NN	O	I-protein
,	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
p27kip1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
establish	NN	O	O
a	NN	O	O
crucial	NN	O	O
PI3K	NN	O	B-protein
/	NN	O	O
PKB	NN	O	B-protein
-mediated	NN	O	O
link	NN	O	O
between	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
teceptor	NN	O	I-protein
and	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
machinery	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
oncogene	NN	O	I-protein
product	NN	O	I-protein
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
engages	NN	O	O
the	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor	NN	O	O
receptor-associated	NN	O	O
death	NN	O	O
domain	NN	O	O
protein	NN	O	O
to	NN	O	O
mediate	NN	O	O
B	NN	O	O
lymphocyte	NN	O	O
growth	NN	O	O
transformation	NN	O	O
and	NN	O	O
activate	NN	O	O
NF-kappaB	NN	O	O
.	NN	O	O

The	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
LMP1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
transformation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
into	NN	O	O
lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Previous	NN	O	O
data	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
model	NN	O	O
that	NN	O	O
LMP1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
constitutively	NN	O	B-protein
activated	NN	O	I-protein
receptor	NN	O	I-protein
that	NN	O	O
transduces	NN	O	O
signals	NN	O	O
for	NN	O	O
transformation	NN	O	O
through	NN	O	O
its	NN	O	O
carboxyl-terminal	NN	O	B-protein
cytoplasmic	NN	O	I-protein
tail	NN	O	I-protein
.	NN	O	O

One	NN	O	O
transformation	NN	O	B-protein
effector	NN	O	I-protein
site	NN	O	I-protein
(	NN	O	O
TES1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
membrane	NN	O	B-protein
proximal	NN	O	I-protein
45	NN	O	I-protein
residues	NN	O	I-protein
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
tail	NN	O	O
,	NN	O	O
constitutively	NN	O	O
engages	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-associated	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Signals	NN	O	O
from	NN	O	O
TES1	NN	O	B-protein
are	NN	O	O
sufficient	NN	O	O
to	NN	O	O
drive	NN	O	O
initial	NN	O	O
proliferation	NN	O	O
of	NN	O	O
infected	NN	O	O
resting	NN	O	O
B	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
most	NN	O	O
lymphoblastoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
infected	NN	O	O
with	NN	O	O
a	NN	O	O
virus	NN	O	O
that	NN	O	O
does	NN	O	O
not	NN	O	O
express	NN	O	O
the	NN	O	O
155	NN	O	B-protein
residues	NN	O	I-protein
beyond	NN	O	O
TES1	NN	O	B-protein
fail	NN	O	O
to	NN	O	O
grow	NN	O	O
as	NN	O	O
long-term	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
now	NN	O	O
find	NN	O	O
that	NN	O	O
mutating	NN	O	O
two	NN	O	O
tyrosines	NN	O	O
to	NN	O	O
an	NN	O	O
isoleucine	NN	O	O
at	NN	O	O
the	NN	O	O
carboxyl	NN	O	B-protein
end	NN	O	I-protein
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
tail	NN	O	I-protein
cripples	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
EBV	NN	O	O
to	NN	O	O
cause	NN	O	O
lymphoblastoid	NN	O	O
cell	NN	O	O
outgrowth	NN	O	O
,	NN	O	O
thereby	NN	O	O
marking	NN	O	O
a	NN	O	O
second	NN	O	O
transformation	NN	O	B-protein
effector	NN	O	I-protein
site	NN	O	I-protein
,	NN	O	O
TES2	NN	O	B-protein
.	NN	O	O

A	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
screen	NN	O	O
identified	NN	O	O
TES2	NN	O	B-protein
interacting	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
including	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-associated	NN	O	I-protein
death	NN	O	I-protein
domain	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
TRADD	NN	O	B-protein
)	NN	O	O
.	NN	O	O

TRADD	NN	O	B-protein
was	NN	O	O
the	NN	O	O
only	NN	O	O
protein	NN	O	O
that	NN	O	O
interacted	NN	O	O
with	NN	O	O
wild-type	NN	O	B-protein
TES2	NN	O	I-protein
and	NN	O	O
not	NN	O	O
with	NN	O	O
isoleucine-mutated	NN	O	B-protein
TES2	NN	O	I-protein
.	NN	O	O

TRADD	NN	O	B-protein
associated	NN	O	O
with	NN	O	O
wild-type	NN	O	B-protein
LMP1	NN	O	I-protein
but	NN	O	O
not	NN	O	O
with	NN	O	O
isoleucine-mutated	NN	O	O
LMP1	NN	O	B-protein
in	NN	O	O
mammalian	NN	O	O
cells	NN	O	O
,	NN	O	O
and	NN	O	O
TRADD	NN	O	B-protein
constitutively	NN	O	O
associated	NN	O	O
with	NN	O	O
LMP1	NN	O	B-protein
in	NN	O	O
EBV-transformed	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
transfection	NN	O	O
assays	NN	O	O
,	NN	O	O
TRADD	NN	O	B-protein
and	NN	O	O
TES2	NN	O	B-protein
synergistically	NN	O	O
mediated	NN	O	O
high-level	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
LMP1	NN	O	B-protein
appropriates	NN	O	O
TRADD	NN	O	B-protein
to	NN	O	O
enable	NN	O	O
efficient	NN	O	O
long-term	NN	O	O
lymphoblastoid	NN	O	O
cell	NN	O	O
outgrowth	NN	O	O
.	NN	O	O

High-level	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
also	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
critical	NN	O	O
component	NN	O	O
of	NN	O	O
long-term	NN	O	O
outgrowth	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
and	NN	O	O
promotion	NN	O	O
of	NN	O	O
natural	NN	O	O
killer	NN	O	O
cell	NN	O	O
development	NN	O	O
by	NN	O	O
the	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
helix	NN	O	I-protein
loop	NN	O	I-protein
helix	NN	O	I-protein
factor	NN	O	I-protein
Id3	NN	O	B-protein
.	NN	O	O

Bipotential	NN	O	B-cell_type
T/natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
thymus	NN	O	O
.	NN	O	O

Despite	NN	O	O
their	NN	O	O
bipotential	NN	O	O
capacity	NN	O	O
,	NN	O	O
these	NN	O	O
progenitors	NN	O	O
develop	NN	O	O
predominantly	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
controlling	NN	O	O
this	NN	O	O
developmental	NN	O	O
choice	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
present	NN	O	O
evidence	NN	O	O
that	NN	O	O
a	NN	O	O
member	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
family	NN	O	O
of	NN	O	O
basic	NN	O	B-protein
helix	NN	O	I-protein
loop	NN	O	I-protein
helix	NN	O	I-protein
(	NN	O	I-protein
bHLH	NN	O	I-protein
)	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
determines	NN	O	O
lineage	NN	O	O
specification	NN	O	O
of	NN	O	O
NK/T	NN	O	B-cell_type
cell	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
natural	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
HLH	NN	O	I-protein
factor	NN	O	I-protein
Id3	NN	O	B-protein
,	NN	O	O
which	NN	O	O
blocks	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
known	NN	O	O
bHLH	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
CD34+	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
retrovirus-mediated	NN	O	O
gene	NN	O	O
transfer	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
Id3	NN	O	B-protein
completely	NN	O	O
blocks	NN	O	O
development	NN	O	O
of	NN	O	O
CD34+	NN	O	B-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
a	NN	O	O
fetal	NN	O	O
thymic	NN	O	O
organ	NN	O	O
culture	NN	O	O
(	NN	O	O
FTOC	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
development	NN	O	O
into	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
an	NN	O	O
FTOC	NN	O	O
is	NN	O	O
enhanced	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
bHLH	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
T	NN	O	O
lineage	NN	O	O
differentiation	NN	O	O
of	NN	O	O
bipotential	NN	O	B-cell_type
precursors	NN	O	I-cell_type
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
which	NN	O	O
a	NN	O	O
default	NN	O	O
pathway	NN	O	O
leading	NN	O	O
to	NN	O	O
NK	NN	O	O
cell	NN	O	O
development	NN	O	O
is	NN	O	O
chosen	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
identify	NN	O	O
a	NN	O	O
molecular	NN	O	O
switch	NN	O	O
for	NN	O	O
lineage	NN	O	O
specification	NN	O	O
in	NN	O	O
early	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
precursors	NN	O	I-cell_type
of	NN	O	O
humans	NN	O	O
.	NN	O	O

-DOCSTART-	O

Late	NN	O	O
gene	NN	O	O
expression	NN	O	O
from	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
BcLF1	NN	O	B-DNA
and	NN	O	I-DNA
BFRF3	NN	O	I-DNA
promoters	NN	O	I-DNA
does	NN	O	O
not	NN	O	O
require	NN	O	O
DNA	NN	O	O
replication	NN	O	O
in	NN	O	O
cis	NN	O	O
.	NN	O	O

Late	NN	O	O
gene	NN	O	O
expression	NN	O	O
follows	NN	O	O
and	NN	O	O
is	NN	O	O
dependent	NN	O	O
upon	NN	O	O
lytic	NN	O	O
replication	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
genome	NN	O	I-DNA
.	NN	O	O

Although	NN	O	O
experimental	NN	O	O
evidence	NN	O	O
is	NN	O	O
lacking	NN	O	O
,	NN	O	O
lytic	NN	O	O
viral	NN	O	O
DNA	NN	O	O
replication	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
remove	NN	O	O
modifications	NN	O	O
or	NN	O	O
binding	NN	O	B-protein
factors	NN	O	I-protein
from	NN	O	O
the	NN	O	O
genome	NN	O	O
which	NN	O	O
serve	NN	O	O
to	NN	O	O
repress	NN	O	O
late	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
latency	NN	O	O
or	NN	O	O
the	NN	O	O
early	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
reporter	NN	O	O
assay	NN	O	O
to	NN	O	O
begin	NN	O	O
characterizing	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
regulate	NN	O	O
late	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
model	NN	O	O
system	NN	O	O
,	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
late	NN	O	B-DNA
promoter-reporter	NN	O	I-DNA
fusions	NN	O	I-DNA
are	NN	O	O
measured	NN	O	O
following	NN	O	O
transient	NN	O	O
transfection	NN	O	O
into	NN	O	O
tissue	NN	O	B-cell_line
culture	NN	O	I-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
EBV	NN	O	O
during	NN	O	O
different	NN	O	O
stages	NN	O	O
of	NN	O	O
the	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
.	NN	O	O

This	NN	O	O
system	NN	O	O
faithfully	NN	O	O
recapitulates	NN	O	O
late	NN	O	O
expression	NN	O	O
patterns	NN	O	O
from	NN	O	O
the	NN	O	O
endogenous	NN	O	O
virus	NN	O	O
,	NN	O	O
implicating	NN	O	O
specific	NN	O	O
cis-active	NN	O	B-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
late	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
these	NN	O	O
promoters	NN	O	O
respond	NN	O	O
only	NN	O	O
indirectly	NN	O	O
to	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
immediate-early	NN	O	I-DNA
transactivator	NN	O	I-DNA
,	NN	O	O
ZEBRA	NN	O	B-DNA
.	NN	O	O

This	NN	O	O
indirect	NN	O	O
response	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
other	NN	O	O
viral	NN	O	O
or	NN	O	O
virally	NN	O	O
induced	NN	O	O
activities	NN	O	O
downstream	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
lytic	NN	O	O
cascade	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
system	NN	O	O
,	NN	O	O
late	NN	O	O
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
sensitive	NN	O	O
to	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
DNA	NN	O	I-protein
polymerase	NN	O	I-protein
such	NN	O	O
as	NN	O	O
phosphonoacetic	NN	O	O
acid	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
reporters	NN	O	O
lack	NN	O	O
a	NN	O	O
eukaryotic	NN	O	O
origin	NN	O	O
of	NN	O	O
replication	NN	O	O
and	NN	O	O
are	NN	O	O
not	NN	O	O
replicated	NN	O	O
under	NN	O	O
the	NN	O	O
assay	NN	O	O
conditions	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
replication	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
template	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
a	NN	O	O
prerequisite	NN	O	O
for	NN	O	O
expression	NN	O	O
with	NN	O	O
late	NN	O	O
kinetics	NN	O	O
,	NN	O	O
a	NN	O	O
finding	NN	O	O
inconsistent	NN	O	O
with	NN	O	O
the	NN	O	O
current	NN	O	O
models	NN	O	O
which	NN	O	O
posit	NN	O	O
a	NN	O	O
cis-active	NN	O	O
relationship	NN	O	O
between	NN	O	O
lytic	NN	O	O
EBV	NN	O	O
DNA	NN	O	O
replication	NN	O	O
and	NN	O	O
late	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
analysis	NN	O	O
of	NN	O	O
this	NN	O	O
system	NN	O	O
has	NN	O	O
revealed	NN	O	O
a	NN	O	O
trans	NN	O	O
relationship	NN	O	O
between	NN	O	O
late	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
viral	NN	O	O
DNA	NN	O	O
replication	NN	O	O
and	NN	O	O
highlights	NN	O	O
the	NN	O	O
indirect	NN	O	O
and	NN	O	O
complex	NN	O	O
link	NN	O	O
between	NN	O	O
these	NN	O	O
two	NN	O	O
events	NN	O	O
.	NN	O	O

-DOCSTART-	O

Modulation	NN	O	O
of	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	B-DNA
human	NN	O	I-DNA
myeloid-selective	NN	O	I-DNA
CCAAT/enhancer	NN	O	I-DNA
binding	NN	O	I-DNA
protein	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
C/EBP	NN	O	B-DNA
epsilon	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

Human	NN	O	O
C/EBP	NN	O	B-DNA
epsilon	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
newly	NN	O	O
cloned	NN	O	O
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
a	NN	O	O
CCAAT/enhancer	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
that	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
levels	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
were	NN	O	O
markedly	NN	O	O
increased	NN	O	O
in	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
promyelocytic	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
because	NN	O	O
they	NN	O	O
were	NN	O	O
induced	NN	O	O
by	NN	O	O
9-cis	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
)	NN	O	O
to	NN	O	O
differentiate	NN	O	O
towards	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

Accumulation	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
occurred	NN	O	O
as	NN	O	O
early	NN	O	O
as	NN	O	O
1	NN	O	O
hour	NN	O	O
after	NN	O	O
exposure	NN	O	O
of	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
(	NN	O	O
5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
mol/L	NN	O	O
)	NN	O	O
;	NN	O	O
and	NN	O	O
at	NN	O	O
48	NN	O	O
hours	NN	O	O
,	NN	O	O
levels	NN	O	O
were	NN	O	O
increased	NN	O	O
by	NN	O	O
5.1-fold	NN	O	O
.	NN	O	O

Dose-response	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
mol/L	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
(	NN	O	O
12	NN	O	O
hours	NN	O	O
)	NN	O	O
resulted	NN	O	O
in	NN	O	O
peak	NN	O	O
levels	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
;	NN	O	O
but	NN	O	O
even	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
mol/L	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
these	NN	O	O
transcripts	NN	O	O
.	NN	O	O

NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
pulse-exposed	NN	O	O
(	NN	O	O
30	NN	O	O
minutes	NN	O	O
)	NN	O	O
to	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
,	NN	O	O
washed	NN	O	O
,	NN	O	O
and	NN	O	O
cultured	NN	O	O
(	NN	O	O
3	NN	O	O
days	NN	O	O
)	NN	O	O
with	NN	O	O
either	NN	O	O
dimethylsulfoxide	NN	O	O
(	NN	O	O
DMSO	NN	O	O
)	NN	O	O
or	NN	O	O
hexamethylene	NN	O	O
bisacetamide	NN	O	O
(	NN	O	O
HMBA	NN	O	O
)	NN	O	O
had	NN	O	O
a	NN	O	O
prominent	NN	O	O
increase	NN	O	O
in	NN	O	O
levels	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
,	NN	O	O
but	NN	O	O
exposure	NN	O	O
to	NN	O	O
either	NN	O	O
DMSO	NN	O	O
or	NN	O	O
HMBA	NN	O	O
alone	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
base	NN	O	O
levels	NN	O	O
of	NN	O	O
C/EBP	NN	O	O
epsilon	NN	O	O
and	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Macrophage-differentiation	NN	O	O
of	NN	O	O
NB4	NN	O	B-cell_line
reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Nuclear	NN	O	O
run-off	NN	O	O
assays	NN	O	O
and	NN	O	O
half-life	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
accumulation	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
enhanced	NN	O	O
transcription	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
this	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
did	NN	O	O
not	NN	O	O
require	NN	O	O
synthesis	NN	O	O
of	NN	O	O
new	NN	O	O
protein	NN	O	B-protein
factors	NN	O	I-protein
because	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
induced	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
new	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

ATRA	NN	O	O
also	NN	O	O
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-protein
epsilon	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
NB4	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
Western	NN	O	O
blotting	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
increase	NN	O	O
of	NN	O	O
C/EBP	NN	O	O
epsilon	NN	O	O
in	NN	O	O
9-cis	NN	O	O
RA-mediated	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
,	NN	O	O
the	NN	O	O
DMSO-induced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
down	NN	O	O
the	NN	O	O
granulocytic	NN	O	O
pathway	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
initial	NN	O	O
reduction	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
discovered	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
markedly	NN	O	O
enhanced	NN	O	O
as	NN	O	O
the	NN	O	O
NB4	NN	O	B-cell_line
promyelocytes	NN	O	I-cell_line
are	NN	O	O
induced	NN	O	O
by	NN	O	O
retinoids	NN	O	O
to	NN	O	O
differentiate	NN	O	O
towards	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

This	NN	O	O
induction	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-RNA
epsilon	NN	O	I-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
is	NN	O	O
transcriptionally	NN	O	O
mediated	NN	O	O
and	NN	O	O
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
synthesis	NN	O	O
of	NN	O	O
additional	NN	O	O
protein	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
suspect	NN	O	O
that	NN	O	O
the	NN	O	O
C/EBP	NN	O	B-DNA
epsilon	NN	O	I-DNA
promoter/enhancer	NN	O	I-DNA
contains	NN	O	O
a	NN	O	O
retinoic	NN	O	B-DNA
acid-response	NN	O	I-DNA
element	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
directly	NN	O	O
stimulated	NN	O	O
by	NN	O	O
retinoids	NN	O	O
.	NN	O	O

-DOCSTART-	O

Dual	NN	O	O
effects	NN	O	O
of	NN	O	O
LPS	NN	O	B-protein
antibodies	NN	O	I-protein
on	NN	O	O
cellular	NN	O	O
uptake	NN	O	O
of	NN	O	O
LPS	NN	O	O
and	NN	O	O
LPS-induced	NN	O	O
proinflammatory	NN	O	O
functions	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
recognize	NN	O	O
bacterial	NN	O	O
LPS	NN	O	O
(	NN	O	O
endotoxin	NN	O	O
)	NN	O	O
through	NN	O	O
membrane	NN	O	B-protein
CD14	NN	O	I-protein
(	NN	O	O
mCD14	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
proinflammatory	NN	O	B-protein
LPS	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

This	NN	O	O
study	NN	O	O
tested	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
anti-LPS	NN	O	B-protein
Abs	NN	O	I-protein
neutralize	NN	O	O
endotoxin	NN	O	O
by	NN	O	O
blocking	NN	O	O
cellular	NN	O	O
uptake	NN	O	O
through	NN	O	O
mCD14	NN	O	B-protein
.	NN	O	O

Ab-associated	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
uptake	NN	O	O
and	NN	O	O
cellular	NN	O	O
distribution	NN	O	O
of	NN	O	O
FITC-LPS	NN	O	O
were	NN	O	O
assessed	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
and	NN	O	O
laser	NN	O	O
scanning	NN	O	O
confocal	NN	O	O
microscopy	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
CD14-transfected	NN	O	I-cell_line
Chinese	NN	O	I-cell_line
hamster	NN	O	I-cell_line
ovary	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
(	NN	O	O
CHO-CD14	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
and	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

LPS	NN	O	O
core-	NN	O	B-protein
and	NN	O	I-protein
O-side	NN	O	I-protein
chain-specific	NN	O	I-protein
mAbs	NN	O	I-protein
inhibited	NN	O	O
mCD14	NN	O	B-protein
-mediated	NN	O	O
LPS	NN	O	O
uptake	NN	O	O
by	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
serum	NN	O	O
.	NN	O	O

O-side	NN	O	B-protein
chain-specific	NN	O	I-protein
mAb	NN	O	I-protein
concurrently	NN	O	O
enhanced	NN	O	O
complement-dependent	NN	O	O
LPS	NN	O	O
uptake	NN	O	O
by	NN	O	O
monocytes	NN	O	O
through	NN	O	O
complement	NN	O	B-protein
receptor-1	NN	O	I-protein
(	NN	O	O
CR1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
uptake	NN	O	O
by	NN	O	O
CHO-CD14	NN	O	B-cell_line
cells	NN	O	I-cell_line
involving	NN	O	O
another	NN	O	O
heat-labile	NN	O	B-protein
serum	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
and	NN	O	O
cell-associated	NN	O	B-protein
recognition	NN	O	I-protein
molecule	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
.	NN	O	O

Core-specific	NN	O	B-protein
mAb	NN	O	I-protein
inhibited	NN	O	O
mCD14	NN	O	B-protein
-mediated	NN	O	O
uptake	NN	O	O
of	NN	O	O
homologous	NN	O	O
and	NN	O	O
heterologous	NN	O	O
LPS	NN	O	O
,	NN	O	O
while	NN	O	O
producing	NN	O	O
less	NN	O	O
concurrent	NN	O	O
enhancement	NN	O	O
of	NN	O	O
non-	NN	O	O
mCD14	NN	O	B-protein
-mediated	NN	O	O
LPS	NN	O	O
uptake	NN	O	O
.	NN	O	O

The	NN	O	O
modulation	NN	O	O
by	NN	O	O
anti-LPS	NN	O	B-protein
mAbs	NN	O	I-protein
of	NN	O	O
mCD14	NN	O	B-protein
-mediated	NN	O	O
LPS	NN	O	O
uptake	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
inhibition	NN	O	O
of	NN	O	O
LPS-induced	NN	O	B-protein
nuclear	NN	O	I-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
translocation	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
secretion	NN	O	O
in	NN	O	O
CHO-CD14	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
monocytes	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
while	NN	O	O
mAb	NN	O	O
enhancement	NN	O	O
of	NN	O	O
non-	NN	O	O
mCD14	NN	O	B-protein
-mediated	NN	O	O
LPS	NN	O	O
uptake	NN	O	O
stimulated	NN	O	O
these	NN	O	O
activities	NN	O	O
.	NN	O	O

LPS-specific	NN	O	B-protein
Abs	NN	O	I-protein
thus	NN	O	O
mediate	NN	O	O
anti-inflammatory	NN	O	O
and	NN	O	O
proinflammatory	NN	O	O
functions	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
by	NN	O	O
preventing	NN	O	O
target	NN	O	O
cell	NN	O	O
uptake	NN	O	O
of	NN	O	O
LPS	NN	O	O
through	NN	O	O
mCD14	NN	O	B-protein
and	NN	O	O
augmenting	NN	O	O
uptake	NN	O	O
through	NN	O	O
CR1	NN	O	B-protein
or	NN	O	O
other	NN	O	O
cell	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Four	NN	O	O
P-like	NN	O	B-DNA
elements	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
IL-4	NN	O	I-DNA
gene	NN	O	I-DNA
:	NN	O	O
involvement	NN	O	O
of	NN	O	O
a	NN	O	O
distinct	NN	O	O
set	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
and	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
previously	NN	O	O
identified	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
critical	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
there	NN	O	O
are	NN	O	O
five	NN	O	O
elements	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
designated	NN	O	O
conserved	NN	O	B-DNA
lymphokine	NN	O	I-DNA
element	NN	O	I-DNA
0	NN	O	I-DNA
(	NN	O	O
CLE0	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
P	NN	O	B-DNA
,	NN	O	O
P2	NN	O	B-DNA
,	NN	O	O
P3	NN	O	B-DNA
and	NN	O	O
P4	NN	O	B-DNA
.	NN	O	O

To	NN	O	O
characterize	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
P-like	NN	O	B-DNA
elements	NN	O	I-DNA
and	NN	O	O
their	NN	O	O
binding	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
native	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
did	NN	O	O
transient	NN	O	O
transfection	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
EL-4	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
promoter-reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
carrying	NN	O	O
mutated	NN	O	B-DNA
P-like	NN	O	I-DNA
elements	NN	O	I-DNA
showed	NN	O	O
that	NN	O	O
four	NN	O	O
P-like	NN	O	B-DNA
elements	NN	O	I-DNA
,	NN	O	O
CLE0	NN	O	B-DNA
,	NN	O	O
P	NN	O	B-DNA
,	NN	O	O
P2	NN	O	B-DNA
and	NN	O	O
P4	NN	O	B-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
P3	NN	O	B-DNA
,	NN	O	O
were	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

EMSA	NN	O	O
showed	NN	O	O
that	NN	O	O
both	NN	O	O
constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
complexes	NN	O	O
bound	NN	O	O
to	NN	O	O
CLE0	NN	O	B-DNA
,	NN	O	O
P	NN	O	B-DNA
,	NN	O	O
P2	NN	O	B-DNA
and	NN	O	O
P4	NN	O	B-DNA
,	NN	O	O
whereas	NN	O	O
only	NN	O	O
a	NN	O	O
constitutive	NN	O	O
complex	NN	O	O
bound	NN	O	O
to	NN	O	O
P3	NN	O	B-DNA
.	NN	O	O

In	NN	O	O
competition	NN	O	O
and	NN	O	O
antibody	NN	O	O
supershift	NN	O	O
assays	NN	O	O
in	NN	O	O
EMSA	NN	O	O
,	NN	O	O
complexes	NN	O	O
formed	NN	O	O
with	NN	O	O
P	NN	O	B-DNA
or	NN	O	O
P2	NN	O	B-DNA
proved	NN	O	O
to	NN	O	O
contain	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	I-protein
NFAT	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
as	NN	O	O
major	NN	O	O
components	NN	O	O
.	NN	O	O

Activator	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
AP	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
interacted	NN	O	O
with	NN	O	O
CLE0	NN	O	B-DNA
,	NN	O	O
P	NN	O	O
,	NN	O	O
P2	NN	O	B-DNA
and	NN	O	O
P4	NN	O	B-DNA
.	NN	O	O

NFAT/AP-1	NN	O	B-protein
complex	NN	O	I-protein
formed	NN	O	O
only	NN	O	O
with	NN	O	O
P	NN	O	B-DNA
and	NN	O	O
P2	NN	O	B-DNA
.	NN	O	O

Cross-competition	NN	O	O
assays	NN	O	O
among	NN	O	O
the	NN	O	O
P-like	NN	O	B-DNA
elements	NN	O	I-DNA
revealed	NN	O	O
element-specific	NN	O	B-protein
and	NN	O	I-protein
common	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Six	NN	O	O
tandem	NN	O	O
repeats	NN	O	O
of	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
element	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
the	NN	O	O
SV40	NN	O	B-DNA
promoter	NN	O	I-DNA
responded	NN	O	O
to	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
,	NN	O	O
while	NN	O	O
that	NN	O	O
of	NN	O	O
other	NN	O	O
elements	NN	O	O
did	NN	O	O
not.	NN	O	O
It	NN	O	O
would	NN	O	O
thus	NN	O	O
appear	NN	O	O
that	NN	O	O
components	NN	O	O
of	NN	O	O
each	NN	O	O
P-like	NN	O	B-protein
element-binding	NN	O	I-protein
complexes	NN	O	I-protein
are	NN	O	O
not	NN	O	O
identical	NN	O	O
and	NN	O	O
may	NN	O	O
coordinately	NN	O	O
contribute	NN	O	O
to	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
expression	NN	O	O
c-fos	NN	O	B-RNA
,	NN	O	I-RNA
c-jun	NN	O	I-RNA
,	NN	O	I-RNA
and	NN	O	I-RNA
NF	NN	O	I-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
in	NN	O	O
nucleated	NN	O	B-cell_type
fetal	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-protein
and	NN	O	O
c-jun	NN	O	B-protein
with	NN	O	O
anti-CD3	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Fetal	NN	O	B-cell_type
and	NN	O	I-cell_type
neonatal	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
are	NN	O	O
relatively	NN	O	O
resistant	NN	O	O
to	NN	O	O
activation	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
when	NN	O	O
stimulated	NN	O	O
either	NN	O	O
via	NN	O	O
their	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptors	NN	O	I-protein
or	NN	O	O
lectins	NN	O	B-protein
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
responsible	NN	O	O
for	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
clearly	NN	O	O
elucidated	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
such	NN	O	O
defects	NN	O	O
in	NN	O	O
fetal/neonatal	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
lack	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
elements	NN	O	I-protein
required	NN	O	O
for	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
to	NN	O	O
examine	NN	O	O
both	NN	O	O
fetal	NN	O	O
and	NN	O	O
term	NN	O	O
neonatal	NN	O	O
cord	NN	O	O
bloods	NN	O	O
for	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
three	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
implicated	NN	O	O
in	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
:	NN	O	O
c-jun	NN	O	B-protein
,	NN	O	O
c-fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
p50	NN	O	B-protein
subunit	NN	O	I-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
mRNAs	NN	O	B-RNA
for	NN	O	O
all	NN	O	O
three	NN	O	O
of	NN	O	O
these	NN	O	O
regulatory	NN	O	O
factors	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
fetal	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
the	NN	O	O
27th	NN	O	O
week	NN	O	O
of	NN	O	O
gestation	NN	O	O
and	NN	O	O
in	NN	O	O
term	NN	O	O
cord	NN	O	O
bloods	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
term	NN	O	B-cell_type
infant	NN	O	I-cell_type
cord	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
anti-CD3	NN	O	B-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
resulted	NN	O	O
in	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
both	NN	O	O
c-jun	NN	O	B-protein
and	NN	O	O
c-fos	NN	O	O
mRNAs	NN	O	B-RNA
within	NN	O	O
15	NN	O	O
min	NN	O	O
of	NN	O	O
stimulation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
by	NN	O	O
anti-CD3-stimulated	NN	O	B-cell_line
cord	NN	O	I-cell_line
blood	NN	O	I-cell_line
mononuclear	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
still	NN	O	O
blunted	NN	O	O
compared	NN	O	O
with	NN	O	O
control	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
adults	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
fetal	NN	O	B-cell_type
nucleated	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
constitutively	NN	O	O
express	NN	O	O
important	NN	O	O
genes	NN	O	O
for	NN	O	O
cytokine	NN	O	B-protein
regulation	NN	O	O
and	NN	O	O
are	NN	O	O
able	NN	O	O
to	NN	O	O
increase	NN	O	O
intracellular	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
mRNAs	NN	O	B-RNA
for	NN	O	O
these	NN	O	O
factors	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
anti-CD3	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
qualitative	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
capacity	NN	O	O
to	NN	O	O
regulate	NN	O	O
these	NN	O	O
factors	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
shown	NN	O	O
in	NN	O	O
fetal	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Quantitative	NN	O	O
experiments	NN	O	O
comparing	NN	O	O
binding	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
are	NN	O	O
currently	NN	O	O
under	NN	O	O
investigation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	O
accumulation	NN	O	O
of	NN	O	O
NFAT4	NN	O	B-protein
opposed	NN	O	O
by	NN	O	O
the	NN	O	O
JNK	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
.	NN	O	O

The	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	I-protein
NFAT	NN	O	I-protein
)	NN	O	I-protein
group	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
is	NN	O	O
retained	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
quiescent	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NFAT	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
mediated	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
induced	NN	O	O
nuclear	NN	O	O
import	NN	O	O
.	NN	O	O

This	NN	O	O
process	NN	O	O
requires	NN	O	O
calcium-dependent	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
caused	NN	O	O
by	NN	O	O
the	NN	O	O
phosphatase	NN	O	B-protein
calcineurin	NN	O	B-protein
.	NN	O	O

The	NN	O	O
c-Jun	NN	O	B-protein
amino-terminal	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
phosphorylates	NN	O	O
NFAT4	NN	O	B-protein
on	NN	O	O
two	NN	O	O
sites	NN	O	O
.	NN	O	O

Mutational	NN	O	O
removal	NN	O	O
of	NN	O	O
the	NN	O	O
JNK	NN	O	B-protein
phosphorylation	NN	O	I-protein
sites	NN	O	I-protein
caused	NN	O	O
constitutive	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
NFAT4	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
calcineurin-stimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
caused	NN	O	O
nuclear	NN	O	O
exclusion	NN	O	O
of	NN	O	O
NFAT4	NN	O	B-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
nuclear	NN	O	O
accumulation	NN	O	O
of	NN	O	O
NFAT4	NN	O	B-protein
promoted	NN	O	O
by	NN	O	O
calcineurin	NN	O	B-protein
is	NN	O	O
opposed	NN	O	O
by	NN	O	O
the	NN	O	O
JNK	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Thiol	NN	O	O
modulation	NN	O	O
inhibits	NN	O	O
the	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
an	NN	O	O
IL-1	NN	O	B-protein
receptor-associated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
interleukin-1	NN	O	B-protein
receptor	NN	O	I-protein
type	NN	O	I-protein
I	NN	O	I-protein
(	NN	O	O
IL-1RI	NN	O	B-protein
)	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
other	NN	O	O
proteins	NN	O	O
thus	NN	O	O
forming	NN	O	O
a	NN	O	O
complex	NN	O	B-protein
system	NN	O	I-protein
by	NN	O	O
which	NN	O	O
IL-1	NN	O	B-protein
exerts	NN	O	O
its	NN	O	O
various	NN	O	O
signals	NN	O	O
.	NN	O	O

The	NN	O	O
initiating	NN	O	O
event	NN	O	O
is	NN	O	O
still	NN	O	O
uncertain	NN	O	O
,	NN	O	O
but	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
recently	NN	O	O
described	NN	O	O
receptor-associated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
earliest	NN	O	O
events	NN	O	O
detectable	NN	O	O
(	NN	O	O
Martin	NN	O	O
et	NN	O	O
al.	NN	O	O
,	NN	O	O
Eur.J.Immunol.1994.24	NN	O	O
:	NN	O	O
1566	NN	O	O
)	NN	O	O
.	NN	O	O

IL-1	NN	O	B-protein
signaling	NN	O	O
is	NN	O	O
commonly	NN	O	O
accompanied	NN	O	O
by	NN	O	O
oxidative	NN	O	O
processes	NN	O	O
and	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
subject	NN	O	O
to	NN	O	O
redox	NN	O	O
regulation	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
investigated	NN	O	O
whether	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1RI-associated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
could	NN	O	O
be	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
redox	NN	O	O
regulation	NN	O	O
and	NN	O	O
whether	NN	O	O
an	NN	O	O
altered	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
kinase	NN	O	O
could	NN	O	O
influence	NN	O	O
IL-1	NN	O	B-protein
-mediated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

A	NN	O	O
murine	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
EL4	NN	O	B-cell_line
,	NN	O	O
was	NN	O	O
stimulated	NN	O	O
with	NN	O	O
IL-1	NN	O	B-protein
with	NN	O	O
and	NN	O	O
without	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
different	NN	O	O
compounds	NN	O	O
known	NN	O	O
to	NN	O	O
influence	NN	O	O
the	NN	O	O
cellular	NN	O	O
redox	NN	O	O
status	NN	O	O
.	NN	O	O

Thiol	NN	O	O
modifying	NN	O	O
agents	NN	O	O
like	NN	O	O
diamide	NN	O	O
,	NN	O	O
menadione	NN	O	O
,	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
,	NN	O	O
diethyl	NN	O	O
dithiocarbamate	NN	O	O
or	NN	O	O
phenylarsine	NN	O	O
oxide	NN	O	O
inhibited	NN	O	O
the	NN	O	O
IL-1	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1RI-associated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

N-Acetylcysteine	NN	O	O
,	NN	O	O
alpha	NN	O	O
,	NN	O	O
alpha'-dipyridyl	NN	O	O
,	NN	O	O
aminotriazole	NN	O	O
or	NN	O	O
nitrofurantoin	NN	O	O
did	NN	O	O
not	NN	O	O
show	NN	O	O
any	NN	O	O
effect	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
by	NN	O	O
PDTC	NN	O	O
was	NN	O	O
reversible	NN	O	O
unless	NN	O	O
glutathione	NN	O	O
synthesis	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
buthionine	NN	O	O
sulfoximine	NN	O	O
.	NN	O	O

The	NN	O	O
described	NN	O	O
conditions	NN	O	O
which	NN	O	O
inhibited	NN	O	O
or	NN	O	O
prevented	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1RI-associated	NN	O	B-protein
kinase	NN	O	I-protein
similarly	NN	O	O
impaired	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
EL4	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

From	NN	O	O
these	NN	O	O
observations	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
free	NN	O	O
thiols	NN	O	O
in	NN	O	O
the	NN	O	O
IL-1RI	NN	O	B-protein
complex	NN	O	I-protein
are	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1RI-associated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
and	NN	O	O
that	NN	O	O
this	NN	O	O
process	NN	O	O
is	NN	O	O
mandatory	NN	O	O
for	NN	O	O
IL-1	NN	O	B-protein
signaling	NN	O	O
leading	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Switching	NN	O	O
gears	NN	O	O
during	NN	O	O
T-cell	NN	O	O
maturation	NN	O	O
:	NN	O	O
RANTES	NN	O	B-protein
and	NN	O	O
late	NN	O	O
transcription	NN	O	O
.	NN	O	O

Although	NN	O	O
much	NN	O	O
is	NN	O	O
understood	NN	O	O
about	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
expressed	NN	O	O
early	NN	O	O
(	NN	O	O
within	NN	O	O
24	NN	O	O
h	NN	O	O
)	NN	O	O
after	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
expressed	NN	O	O
'late	NN	O	O
'	NN	O	O
(	NN	O	O
three	NN	O	O
or	NN	O	O
more	NN	O	O
days	NN	O	O
)	NN	O	O
post-stimulation	NN	O	O
.	NN	O	O

A	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
at	NN	O	O
this	NN	O	O
important	NN	O	O
stage	NN	O	O
of	NN	O	O
T-cell	NN	O	O
maturation	NN	O	O
may	NN	O	O
yield	NN	O	O
new	NN	O	O
insights	NN	O	O
into	NN	O	O
T-cell	NN	O	O
development	NN	O	O
and	NN	O	O
new	NN	O	O
immunotherapeutic	NN	O	O
targets	NN	O	O
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
different	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
several	NN	O	O
inducers	NN	O	B-DNA
.	NN	O	O

Double-stimulation	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
T	NN	O	B-cell_line
lymphocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
CEM	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
through	NN	O	O
a	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
which	NN	O	O
did	NN	O	O
not	NN	O	O
involve	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
and	NN	O	O
was	NN	O	O
different	NN	O	O
from	NN	O	O
the	NN	O	O
activation	NN	O	O
triggered	NN	O	O
by	NN	O	O
either	NN	O	O
H2O2	NN	O	O
or	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Since	NN	O	O
these	NN	O	O
latter	NN	O	O
compounds	NN	O	O
were	NN	O	O
known	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
in	NN	O	O
a	NN	O	O
redox-sensitive	NN	O	O
way	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
PMA	NN	O	O
was	NN	O	O
resistant	NN	O	O
to	NN	O	O
antioxidant	NN	O	O
N-acetyl-L-cysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
and	NN	O	O
sensitive	NN	O	O
to	NN	O	O
kinase	NN	O	O
inhibitors	NN	O	O
staurosporine	NN	O	O
and	NN	O	O
H7	NN	O	O
while	NN	O	O
activation	NN	O	O
by	NN	O	O
H2O2	NN	O	O
or	NN	O	O
TNF-alpha	NN	O	B-protein
were	NN	O	O
not	NN	O	O
.	NN	O	O

-DOCSTART-	O

Blockade	NN	O	O
of	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
by	NN	O	O
dithiocarbamates	NN	O	O
involves	NN	O	O
novel	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
inhibition	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
.	NN	O	O

Dithiocarbamates	NN	O	O
(	NN	O	O
DTCs	NN	O	O
)	NN	O	O
have	NN	O	O
recently	NN	O	O
been	NN	O	O
reported	NN	O	O
as	NN	O	O
powerful	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Given	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
transcription	NN	O	O
factor	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
inhibitors	NN	O	O
have	NN	O	O
been	NN	O	O
suggested	NN	O	O
as	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
drugs	NN	O	O
for	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
DTCs	NN	O	O
inhibited	NN	O	O
both	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
synthesis	NN	O	O
and	NN	O	O
membrane	NN	O	O
expression	NN	O	O
of	NN	O	O
antigens	NN	O	B-protein
which	NN	O	O
are	NN	O	O
induced	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
,	NN	O	O
which	NN	O	O
occurred	NN	O	O
with	NN	O	O
a	NN	O	O
parallel	NN	O	O
activation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
transactivating	NN	O	O
functions	NN	O	O
and	NN	O	O
expression	NN	O	O
,	NN	O	O
was	NN	O	O
reflected	NN	O	O
by	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
at	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
level	NN	O	O
,	NN	O	O
and	NN	O	O
involved	NN	O	O
not	NN	O	O
only	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
-driven	NN	O	O
reporter	NN	O	O
activation	NN	O	O
but	NN	O	O
also	NN	O	O
that	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Accordingly	NN	O	O
,	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSAs	NN	O	O
)	NN	O	O
indicated	NN	O	O
that	NN	O	O
pyrrolidine	NN	O	O
DTC	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
prevented	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NFAT	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
either	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
plus	NN	O	O
ionophore	NN	O	O
or	NN	O	O
antibodies	NN	O	O
against	NN	O	O
the	NN	O	O
CD3-T-cell	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
and	NN	O	O
simultaneously	NN	O	O
activated	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
PDTC	NN	O	O
differentially	NN	O	O
targeted	NN	O	O
both	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	I-protein
NFATc	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
,	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
transactivation	NN	O	O
functions	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	O
mRNA	NN	O	O
induction	NN	O	O
of	NN	O	O
NFATc	NN	O	B-protein
.	NN	O	O

Strikingly	NN	O	O
,	NN	O	O
Western	NN	O	O
blotting	NN	O	O
and	NN	O	O
immunocytochemical	NN	O	O
experiments	NN	O	O
indicated	NN	O	O
that	NN	O	O
PDTC	NN	O	O
promoted	NN	O	O
a	NN	O	O
transient	NN	O	O
and	NN	O	O
rapid	NN	O	O
shuttling	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	O
NFATc	NN	O	B-protein
,	NN	O	O
leading	NN	O	O
to	NN	O	O
their	NN	O	O
accelerated	NN	O	O
export	NN	O	O
from	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
an	NN	O	O
NFAT	NN	O	O
kinase	NN	O	O
by	NN	O	O
PDTC	NN	O	O
could	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
rapid	NN	O	O
shuttling	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
therefore	NN	O	O
transiently	NN	O	O
converting	NN	O	O
the	NN	O	O
sustained	NN	O	O
transactivation	NN	O	O
of	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
occurs	NN	O	O
during	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
show	NN	O	O
that	NN	O	O
c-Jun	NN	O	B-protein
NH2-terminal	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
can	NN	O	O
act	NN	O	O
by	NN	O	O
directly	NN	O	O
phosphorylating	NN	O	O
NFATp	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
combined	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
on	NN	O	O
NFAT	NN	O	B-protein
and	NN	O	O
NF-KB	NN	O	B-protein
support	NN	O	O
a	NN	O	O
potential	NN	O	O
use	NN	O	O
of	NN	O	O
DTCs	NN	O	O
as	NN	O	O
immunosuppressants	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
3q21	NN	O	B-DNA
and	NN	O	O
identification	NN	O	O
of	NN	O	O
intergenic	NN	O	O
fusion	NN	O	O
transcripts	NN	O	O
with	NN	O	O
ecotropic	NN	O	B-DNA
viral	NN	O	I-DNA
insertion	NN	O	I-DNA
site	NN	O	I-DNA
I	NN	O	I-DNA
in	NN	O	O
leukemia	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
GR6	NN	O	B-DNA
,	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
leukemia	NN	O	O
breakpoint	NN	O	O
region	NN	O	O
of	NN	O	O
3q21	NN	O	B-DNA
,	NN	O	O
that	NN	O	O
is	NN	O	O
normally	NN	O	O
expressed	NN	O	O
in	NN	O	O
early	NN	O	O
fetal	NN	O	O
development	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
adult	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
.	NN	O	O

GR6	NN	O	B-DNA
is	NN	O	O
activated	NN	O	O
in	NN	O	O
the	NN	O	O
UCSD-AML1	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
a	NN	O	O
leukemic	NN	O	B-cell_line
sample	NN	O	I-cell_line
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
carry	NN	O	O
a	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
3	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q21	NN	O	I-DNA
;	NN	O	I-DNA
q26	NN	O	I-DNA
)	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
UCSD-AML1	NN	O	B-cell_line
,	NN	O	O
we	NN	O	O
have	NN	O	O
also	NN	O	O
identified	NN	O	O
fusion	NN	O	B-RNA
transcripts	NN	O	I-RNA
between	NN	O	O
the	NN	O	O
ecotropic	NN	O	B-DNA
viral	NN	O	I-DNA
insertion	NN	O	I-DNA
site	NN	O	I-DNA
I	NN	O	I-DNA
(	NN	O	I-DNA
EVI1	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
3q26	NN	O	B-DNA
and	NN	O	O
GR6	NN	O	B-DNA
and	NN	O	O
between	NN	O	O
EVI1	NN	O	B-DNA
and	NN	O	O
Ribophorin	NN	O	B-DNA
I	NN	O	I-DNA
that	NN	O	O
maps	NN	O	O
30	NN	O	O
kb	NN	O	O
telomeric	NN	O	O
to	NN	O	O
GR6	NN	O	B-DNA
in	NN	O	O
3q21	NN	O	B-DNA
.	NN	O	O

All	NN	O	O
fusions	NN	O	O
splice	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
ends	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
3q21	NN	O	B-DNA
genes	NN	O	I-DNA
into	NN	O	O
exon	NN	O	B-DNA
2	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
EVI1	NN	O	B-DNA
gene	NN	O	O
,	NN	O	O
an	NN	O	O
event	NN	O	O
that	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
normal	NN	O	O
intergenic	NN	O	O
splicing	NN	O	O
of	NN	O	O
MDS1-EVI1	NN	O	B-DNA
and	NN	O	O
to	NN	O	O
those	NN	O	O
previously	NN	O	O
documented	NN	O	O
in	NN	O	O
leukemias	NN	O	O
with	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
21	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
12	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
which	NN	O	O
acute	NN	O	O
myelogenous	NN	O	B-DNA
leukemia	NN	O	I-DNA
1-EVI1	NN	O	I-DNA
fusions	NN	O	I-DNA
and	NN	O	O
ETV6-EVI1	NN	O	B-DNA
fusions	NN	O	I-DNA
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
occur	NN	O	O
.	NN	O	O

The	NN	O	O
Ribophorin	NN	O	B-DNA
I-EVI1	NN	O	I-DNA
fusion	NN	O	I-DNA
in	NN	O	O
particular	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
common	NN	O	O
occurrence	NN	O	O
in	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
3	NN	O	I-DNA
)	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
A-myb	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
neoplastic	NN	O	B-cell_type
and	NN	O	I-cell_type
normal	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
myb	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
has	NN	O	O
been	NN	O	O
strongly	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
in	NN	O	O
the	NN	O	O
haematopoietic	NN	O	O
system	NN	O	O
.	NN	O	O

The	NN	O	O
v-myb	NN	O	B-DNA
oncogene	NN	O	I-DNA
,	NN	O	O
carried	NN	O	O
by	NN	O	O
avian	NN	O	O
defective	NN	O	O
retroviruses	NN	O	O
,	NN	O	O
causes	NN	O	O
leukaemias	NN	O	O
in	NN	O	O
the	NN	O	O
chicken	NN	O	O
and	NN	O	O
transforms	NN	O	O
haematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Its	NN	O	O
normal	NN	O	O
cellular	NN	O	O
equivalent	NN	O	O
c-myb	NN	O	B-DNA
,	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
promote	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and	NN	O	O
block	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
haematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
several	NN	O	O
experimental	NN	O	O
models	NN	O	O
and	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
fetal	NN	O	O
haematopoiesis	NN	O	O
.	NN	O	O

Two	NN	O	O
other	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
family	NN	O	O
have	NN	O	O
been	NN	O	O
cloned	NN	O	O
more	NN	O	O
recently	NN	O	O
,	NN	O	O
A-myb	NN	O	B-protein
and	NN	O	O
B-myb	NN	O	B-protein
,	NN	O	O
which	NN	O	O
show	NN	O	O
sequence	NN	O	O
homology	NN	O	O
with	NN	O	O
c-myb	NN	O	B-DNA
in	NN	O	O
several	NN	O	O
domains	NN	O	O
,	NN	O	O
of	NN	O	O
which	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
other	NN	O	O
regulatory	NN	O	B-protein
domains	NN	O	I-protein
.	NN	O	O

Both	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

B-myb	NN	O	B-protein
is	NN	O	O
also	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
,	NN	O	O
but	NN	O	O
,	NN	O	O
unlike	NN	O	O
c-myb	NN	O	B-DNA
,	NN	O	O
it	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
many	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

The	NN	O	O
third	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
family	NN	O	B-protein
,	NN	O	O
A-myb	NN	O	B-protein
,	NN	O	O
shows	NN	O	O
the	NN	O	O
most	NN	O	O
restricted	NN	O	O
pattern	NN	O	O
of	NN	O	O
expression	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
very	NN	O	O
specific	NN	O	O
role	NN	O	O
for	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

A-myb	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
subpopulation	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
activated	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
localised	NN	O	O
in	NN	O	O
the	NN	O	O
germinal	NN	O	O
center	NN	O	O
of	NN	O	O
peripheral	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
detected	NN	O	O
at	NN	O	O
significant	NN	O	O
levels	NN	O	O
in	NN	O	O
all	NN	O	O
other	NN	O	O
mature	NN	O	O
or	NN	O	O
immature	NN	O	B-cell_type
haematopoietic	NN	O	I-cell_type
populations	NN	O	I-cell_type
studied	NN	O	O
,	NN	O	O
including	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
granulocytes	NN	O	B-cell_type
,	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
either	NN	O	O
at	NN	O	O
rest	NN	O	O
or	NN	O	O
after	NN	O	O
in	NN	O	O
vitro	NN	O	O
activation	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
A-myb	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
during	NN	O	O
a	NN	O	O
narrow	NN	O	O
window	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
B	NN	O	I-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
.	NN	O	O

A-myb	NN	O	B-protein
expression	NN	O	O
has	NN	O	O
also	NN	O	O
been	NN	O	O
studied	NN	O	O
in	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
neoplastic	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
representing	NN	O	O
the	NN	O	O
whole	NN	O	O
spectrum	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

A-myb	NN	O	B-protein
is	NN	O	O
strongly	NN	O	O
expressed	NN	O	O
in	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphomas	NN	O	O
(	NN	O	O
BL	NN	O	O
)	NN	O	O
and	NN	O	O
slg+	NN	O	O
B-acute	NN	O	O
lymphoblastic	NN	O	O
leukaemias	NN	O	O
(	NN	O	O
B-ALL	NN	O	O
)	NN	O	O
and	NN	O	O
not	NN	O	O
in	NN	O	O
all	NN	O	O
other	NN	O	O
leukaemias/lymphomas	NN	O	O
tested	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
CLL	NN	O	O
(	NN	O	O
about	NN	O	O
25	NN	O	O
%	NN	O	O
of	NN	O	O
cases	NN	O	O
)	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
intriguing	NN	O	O
that	NN	O	O
the	NN	O	O
A-myb	NN	O	B-DNA
genome	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
localised	NN	O	O
relatively	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
c-myc	NN	O	B-DNA
gene	NN	O	I-DNA
on	NN	O	O
chromosome	NN	O	B-DNA
8	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
c-myc	NN	O	O
translocation	NN	O	O
in	NN	O	O
BL	NN	O	O
and	NN	O	O
B-ALL	NN	O	O
may	NN	O	O
affect	NN	O	O
A-myb	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

Studies	NN	O	O
are	NN	O	O
in	NN	O	O
progress	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
functional	NN	O	O
relationship	NN	O	O
between	NN	O	O
A-myb	NN	O	B-protein
and	NN	O	O
c-myc	NN	O	B-DNA
,	NN	O	O
particularly	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
BL	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
A-myb	NN	O	B-protein
is	NN	O	O
deregulated	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
bcl-6	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
normal	NN	O	O
skin	NN	O	O
and	NN	O	O
epidermal	NN	O	O
neoplasms	NN	O	O
.	NN	O	O

Bcl-6	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
a	NN	O	O
recently	NN	O	O
identified	NN	O	O
novel	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
whose	NN	O	O
deregulated	NN	O	O
expression	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
diffuse	NN	O	O
large	NN	O	O
B	NN	O	O
cell	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
recently	NN	O	O
shown	NN	O	O
by	NN	O	O
us	NN	O	O
that	NN	O	O
the	NN	O	O
protein	NN	O	O
is	NN	O	O
located	NN	O	O
in	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
their	NN	O	O
neoplastic	NN	O	B-cell_type
counterparts	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
bcl-6	NN	O	B-protein
protein	NN	O	I-protein
on	NN	O	O
normal	NN	O	O
epidermis	NN	O	O
,	NN	O	O
benign	NN	O	O
,	NN	O	O
and	NN	O	O
malignant	NN	O	O
tumors	NN	O	O
originating	NN	O	O
from	NN	O	O
epidermal	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
squamous	NN	O	B-cell_line
cell	NN	O	I-cell_line
carcinoma	NN	O	I-cell_line
(	NN	O	I-cell_line
SCC	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
are	NN	O	O
investigated	NN	O	O
.	NN	O	O

With	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
immunohistochemistry	NN	O	O
,	NN	O	O
bcl-6	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
stain	NN	O	O
intensely	NN	O	O
on	NN	O	O
normal	NN	O	B-cell_line
prickle	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
none	NN	O	O
to	NN	O	O
only	NN	O	O
slightly	NN	O	O
on	NN	O	O
epidermal	NN	O	B-cell_type
basal	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Papillomas	NN	O	O
and	NN	O	O
keratoacanthomas	NN	O	O
copied	NN	O	O
their	NN	O	O
normal	NN	O	O
counterparts	NN	O	O
in	NN	O	O
the	NN	O	O
mode	NN	O	O
of	NN	O	O
expression	NN	O	O
.	NN	O	O

Various	NN	O	O
levels	NN	O	O
of	NN	O	O
expression	NN	O	O
were	NN	O	O
found	NN	O	O
on	NN	O	O
seborrheic	NN	O	O
keratoses	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
expression	NN	O	O
level	NN	O	O
on	NN	O	O
basal	NN	O	O
cell	NN	O	O
epitheliomas	NN	O	O
was	NN	O	O
low	NN	O	O
.	NN	O	O

Peculiarly	NN	O	O
,	NN	O	O
eccrine	NN	O	O
poromas	NN	O	O
and	NN	O	O
undifferentiated	NN	O	O
spindle-shaped	NN	O	O
basal	NN	O	O
cell	NN	O	O
epitheliomas	NN	O	O
were	NN	O	O
totally	NN	O	O
unstained	NN	O	O
.	NN	O	O

Squamous	NN	O	O
cell	NN	O	O
carcinomas	NN	O	O
showed	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
expression	NN	O	O
levels	NN	O	O
,	NN	O	O
while	NN	O	O
two	NN	O	O
undifferentiated	NN	O	O
spindle-shaped	NN	O	O
carcinomas	NN	O	O
and	NN	O	O
one	NN	O	O
undifferentiated	NN	O	O
SCC	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
remained	NN	O	O
unstained	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
bcl-6	NN	O	B-protein
protein	NN	O	I-protein
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
morphological	NN	O	O
differentiation	NN	O	O
in	NN	O	O
normal	NN	O	O
and	NN	O	O
neoplastic	NN	O	B-cell_type
epidermal	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Sp3	NN	O	B-protein
mediates	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
leukocyte	NN	O	B-DNA
integrin	NN	O	I-DNA
genes	NN	O	I-DNA
CD11C	NN	O	B-DNA
and	NN	O	O
CD11B	NN	O	B-DNA
and	NN	O	O
cooperates	NN	O	O
with	NN	O	O
c-Jun	NN	O	O
to	NN	O	O
activate	NN	O	O
CD11C	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
leukocyte	NN	O	B-DNA
integrin	NN	O	I-DNA
genes	NN	O	I-DNA
CD11c	NN	O	B-DNA
and	NN	O	O
CD11b	NN	O	B-DNA
are	NN	O	O
expressed	NN	O	O
predominately	NN	O	O
in	NN	O	O
myelomonocytic	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
previous	NN	O	O
experiments	NN	O	O
,	NN	O	O
the	NN	O	O
-70	NN	O	B-DNA
to	NN	O	I-DNA
-65	NN	O	I-DNA
and	NN	O	I-DNA
-121	NN	O	I-DNA
to	NN	O	I-DNA
-103	NN	O	I-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CD11c	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
-66	NN	O	B-DNA
to	NN	O	I-DNA
-59	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
promoter	NN	O	I-DNA
were	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
essential	NN	O	O
for	NN	O	O
Sp1-	NN	O	B-protein
mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
genomic	NN	O	O
footprinting	NN	O	O
had	NN	O	O
also	NN	O	O
revealed	NN	O	O
cell-specific	NN	O	O
binding	NN	O	O
of	NN	O	O
protein	NN	O	O
,	NN	O	O
presumably	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
to	NN	O	O
these	NN	O	O
regions	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
Sp1-related	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
Sp3	NN	O	B-protein
,	NN	O	O
also	NN	O	O
binds	NN	O	O
at	NN	O	O
or	NN	O	O
near	NN	O	O
these	NN	O	O
same	NN	O	O
regions	NN	O	O
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
Sp3	NN	O	B-protein
along	NN	O	O
with	NN	O	O
CD11c	NN	O	B-DNA
promoter-luciferase	NN	O	I-DNA
constructs	NN	O	I-DNA
into	NN	O	O
Sp-deficient	NN	O	B-cell_line
Drosophila	NN	O	I-cell_line
Schneider	NN	O	I-cell_line
2	NN	O	I-cell_line
cells	NN	O	I-cell_line
showed	NN	O	O
that	NN	O	O
Sp3	NN	O	B-protein
could	NN	O	O
activate	NN	O	O
the	NN	O	O
CD11c	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
-70	NN	O	B-DNA
to	NN	O	I-DNA
-65	NN	O	I-DNA
and	NN	O	I-DNA
-121	NN	O	I-DNA
to	NN	O	I-DNA
-103	NN	O	I-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CD11c	NN	O	B-DNA
promoter	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
activation	NN	O	O
by	NN	O	O
Sp3	NN	O	B-protein
.	NN	O	O

Both	NN	O	O
sites	NN	O	O
showed	NN	O	O
activation	NN	O	O
by	NN	O	O
Sp3	NN	O	B-protein
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
-70	NN	O	B-DNA
to	NN	O	I-DNA
-65	NN	O	I-DNA
region	NN	O	I-DNA
was	NN	O	O
more	NN	O	O
responsive	NN	O	O
to	NN	O	O
Sp3	NN	O	B-protein
than	NN	O	O
to	NN	O	O
Sp1	NN	O	B-protein
.	NN	O	O

Similar	NN	O	O
transfection	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
-66	NN	O	B-DNA
to	NN	O	I-DNA
-59	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
promoter	NN	O	I-DNA
showed	NN	O	O
Sp3	NN	O	B-protein
-dependent	NN	O	O
expression	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
cotransfection	NN	O	O
analysis	NN	O	O
in	NN	O	O
Drosophila	NN	O	B-cell_line
cells	NN	O	I-cell_line
showed	NN	O	O
that	NN	O	O
Sp3	NN	O	B-protein
,	NN	O	O
as	NN	O	O
was	NN	O	O
previously	NN	O	O
shown	NN	O	O
for	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
also	NN	O	O
synergizes	NN	O	O
with	NN	O	O
c-Jun	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
CD11c	NN	O	B-DNA
.	NN	O	O

Antisense	NN	O	O
experiments	NN	O	O
that	NN	O	O
knocked	NN	O	O
out	NN	O	O
endogenous	NN	O	O
Sp3	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
the	NN	O	O
myelomocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
HL60	NN	O	B-cell_line
,	NN	O	O
revealed	NN	O	O
that	NN	O	O
Sp3	NN	O	B-protein
participates	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
CD11c	NN	O	B-DNA
and	NN	O	I-DNA
CD11b	NN	O	I-DNA
promoters	NN	O	I-DNA
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
winged-helix	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Trident	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
actively	NN	O	O
dividing	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
recently	NN	O	O
identified	NN	O	O
the	NN	O	O
winged-helix	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Trident	NN	O	B-protein
and	NN	O	O
described	NN	O	O
its	NN	O	O
expression	NN	O	O
pattern	NN	O	O
in	NN	O	O
synchronized	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
studied	NN	O	O
Trident	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
differentiating	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
lymphocytes	NN	O	O
derived	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
.	NN	O	O

During	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
expression	NN	O	O
peaked	NN	O	O
in	NN	O	O
the	NN	O	O
actively	NN	O	B-cell_type
dividing	NN	O	I-cell_type
immature	NN	O	I-cell_type
single	NN	O	I-cell_type
positive	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
lymphocytes	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
Trident	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
absent	NN	O	O
,	NN	O	O
but	NN	O	O
could	NN	O	O
be	NN	O	O
induced	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
mitogens	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
imply	NN	O	O
a	NN	O	O
function	NN	O	O
for	NN	O	O
Trident	NN	O	B-protein
in	NN	O	O
dividing	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Estrogen	NN	O	B-protein
receptor	NN	O	I-protein
diminishes	NN	O	O
DNA-binding	NN	O	O
activities	NN	O	O
of	NN	O	O
chicken	NN	O	B-protein
GATA-1	NN	O	I-protein
and	NN	O	O
CACCC-binding	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
repressed	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
and	NN	O	O
erythroid-specific	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
alpha	NN	O	I-protein
(	NN	O	O
referred	NN	O	O
to	NN	O	O
throughout	NN	O	O
as	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
on	NN	O	O
DNA-binding	NN	O	O
activities	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
erythroid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
the	NN	O	O
histone	NN	O	B-DNA
H5	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
rabbit	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
,	NN	O	O
human	NN	O	B-protein
ER	NN	O	I-protein
reduced	NN	O	O
the	NN	O	O
binding	NN	O	O
activities	NN	O	O
of	NN	O	O
chicken	NN	O	B-protein
immature	NN	O	I-protein
erythrocyte	NN	O	I-protein
nuclear	NN	O	I-protein
extracted	NN	O	I-protein
proteins	NN	O	I-protein
to	NN	O	O
GATA	NN	O	B-DNA
and	NN	O	I-DNA
CACCC	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
H5	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
activities	NN	O	O
of	NN	O	O
NF1	NN	O	B-protein
and	NN	O	O
Sp1	NN	O	B-protein
were	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
ER	NN	O	B-protein
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
to	NN	O	O
an	NN	O	O
estrogen	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
was	NN	O	O
enhanced	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
rabbit	NN	O	O
reticulocyte	NN	O	O
lysate	NN	O	O
.	NN	O	O

This	NN	O	O
lysate	NN	O	O
was	NN	O	O
also	NN	O	O
necessary	NN	O	O
for	NN	O	O
ER	NN	O	B-protein
to	NN	O	O
diminish	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
additional	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
are	NN	O	O
necessary	NN	O	O
for	NN	O	O
full	NN	O	O
ER	NN	O	B-protein
function	NN	O	O
.	NN	O	O

Both	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
CACCC-binding	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
developmentally	NN	O	O
regulated	NN	O	O
expression	NN	O	O
of	NN	O	O
erythroid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
interference	NN	O	O
in	NN	O	O
DNA-binding	NN	O	O
activities	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
CACCC-binding	NN	O	B-protein
proteins	NN	O	I-protein
is	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
the	NN	O	O
ER	NN	O	B-protein
inhibits	NN	O	O
regulation	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
Id3	NN	O	B-protein
cell	NN	O	O
cycle	NN	O	O
function	NN	O	O
by	NN	O	O
Cdk-2-dependent	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

The	NN	O	O
functions	NN	O	O
of	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
(	NN	O	I-protein
bHLH	NN	O	I-protein
)	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
activating	NN	O	O
differentiation-linked	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
in	NN	O	O
inducing	NN	O	O
G1	NN	O	O
cell	NN	O	O
cycle	NN	O	O
arrest	NN	O	O
are	NN	O	O
negatively	NN	O	O
regulated	NN	O	O
by	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Id	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
HLH	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

These	NN	O	O
bHLH	NN	O	B-protein
antagonists	NN	O	I-protein
are	NN	O	O
induced	NN	O	O
during	NN	O	O
a	NN	O	O
mitogenic	NN	O	O
signalling	NN	O	O
response	NN	O	O
,	NN	O	O
and	NN	O	O
they	NN	O	O
function	NN	O	O
by	NN	O	O
sequestering	NN	O	O
their	NN	O	O
bHLH	NN	O	B-protein
targets	NN	O	I-protein
in	NN	O	O
inactive	NN	O	B-protein
heterodimers	NN	O	I-protein
that	NN	O	O
are	NN	O	O
unable	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
specific	NN	O	O
gene	NN	O	B-DNA
regulatory	NN	O	I-DNA
(	NN	O	I-DNA
E	NN	O	I-DNA
box	NN	O	I-DNA
)	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
cyclin	NN	O	O
E-Cdk2-	NN	O	O
and	NN	O	O
cyclin	NN	O	O
A-Cdk2-dependent	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
conserved	NN	O	O
serine	NN	O	O
residue	NN	O	O
(	NN	O	O
Ser5	NN	O	O
)	NN	O	O
in	NN	O	O
Id2	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
occur	NN	O	O
during	NN	O	O
late	NN	O	O
G1-to-S	NN	O	O
phase	NN	O	O
transition	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
neutralizes	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
Id2	NN	O	B-protein
in	NN	O	O
abrogating	NN	O	O
E-box-dependent	NN	O	B-protein
bHLH	NN	O	I-protein
homo	NN	O	O
-or	NN	O	O
heterodimer	NN	O	O
complex	NN	O	O
formation	NN	O	O
in	NN	O	O
vitro	NN	O	O
(	NN	O	O
E.Hara	NN	O	O
,	NN	O	O
M.Hall	NN	O	O
,	NN	O	O
and	NN	O	O
G.Peters	NN	O	O
,	NN	O	O
EMBO	NN	O	O
J.16	NN	O	O
:	NN	O	O
332-342	NN	O	O
,	NN	O	O
1997	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
an	NN	O	O
analogous	NN	O	O
cell-cycle-regulated	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Id3	NN	O	B-protein
alters	NN	O	O
the	NN	O	O
specificity	NN	O	O
of	NN	O	O
Id3	NN	O	B-protein
for	NN	O	O
abrogating	NN	O	O
both	NN	O	O
E-box-dependent	NN	O	B-protein
bHLH	NN	O	I-protein
homo-	NN	O	O
or	NN	O	O
heterodimer	NN	O	O
complex	NN	O	O
formation	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
E-box-dependent	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
function	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
compared	NN	O	O
with	NN	O	O
wild-type	NN	O	B-protein
Id3	NN	O	I-protein
,	NN	O	O
an	NN	O	O
Id3	NN	O	B-protein
Asp5	NN	O	I-protein
mutant	NN	O	I-protein
(	NN	O	O
mimicking	NN	O	O
phosphorylation	NN	O	O
)	NN	O	O
is	NN	O	O
unable	NN	O	O
to	NN	O	O
promote	NN	O	O
cell	NN	O	O
cycle	NN	O	O
S	NN	O	O
phase	NN	O	O
entry	NN	O	O
in	NN	O	O
transfected	NN	O	B-cell_line
fibroblasts	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
an	NN	O	O
Id3	NN	O	B-protein
Ala5	NN	O	I-protein
mutant	NN	O	I-protein
(	NN	O	O
ablating	NN	O	O
phosphorylation	NN	O	O
)	NN	O	O
displays	NN	O	O
an	NN	O	O
activity	NN	O	O
significantly	NN	O	O
greater	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
wild-type	NN	O	B-protein
Id3	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Cdk2-dependent	NN	O	O
phosphorylation	NN	O	O
therefore	NN	O	O
provides	NN	O	O
a	NN	O	O
switch	NN	O	O
during	NN	O	O
late	NN	O	O
G1-to-S	NN	O	O
phase	NN	O	O
that	NN	O	O
both	NN	O	O
nullifies	NN	O	O
an	NN	O	O
early	NN	O	O
G1	NN	O	O
cell	NN	O	O
cycle	NN	O	O
regulatory	NN	O	O
function	NN	O	O
of	NN	O	O
Id3	NN	O	B-protein
and	NN	O	O
modulates	NN	O	O
its	NN	O	O
target	NN	O	O
bHLH	NN	O	B-protein
specificity	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
Id3	NN	O	B-protein
to	NN	O	O
promote	NN	O	O
cell	NN	O	O
cycle	NN	O	O
S	NN	O	O
phase	NN	O	O
entry	NN	O	O
is	NN	O	O
not	NN	O	O
simply	NN	O	O
a	NN	O	O
function	NN	O	O
of	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
modulate	NN	O	O
bHLH	NN	O	B-protein
heterodimer-dependent	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
establish	NN	O	O
a	NN	O	O
biologically	NN	O	O
important	NN	O	O
mechanism	NN	O	O
through	NN	O	O
which	NN	O	O
Cdk2	NN	O	O
and	NN	O	O
Id-bHLH	NN	O	O
functions	NN	O	O
are	NN	O	O
integrated	NN	O	O
in	NN	O	O
the	NN	O	O
coordination	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
cloning	NN	O	O
and	NN	O	O
functional	NN	O	O
characterization	NN	O	O
of	NN	O	O
murine	NN	O	B-protein
cDNA	NN	O	I-protein
encoding	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NFATc	NN	O	B-protein
.	NN	O	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
(	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
play	NN	O	O
important	NN	O	O
roles	NN	O	O
in	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
by	NN	O	O
regulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
encoding	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
immunoregulatory	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
cloning	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
full-length	NN	O	O
cDNA	NN	O	B-protein
encoding	NN	O	I-protein
murine	NN	O	I-protein
(	NN	O	I-protein
m	NN	O	I-protein
)	NN	O	I-protein
NFATc	NN	O	I-protein
which	NN	O	O
predicts	NN	O	O
that	NN	O	O
the	NN	O	O
protein	NN	O	O
has	NN	O	O
all	NN	O	O
the	NN	O	O
conserved	NN	O	B-protein
structural	NN	O	I-protein
motifs	NN	O	I-protein
of	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
,	NN	O	O
including	NN	O	O
the	NN	O	O
rel	NN	O	O
homology	NN	O	O
domain	NN	O	O
,	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
homology	NN	O	I-protein
domain	NN	O	I-protein
and	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
signals	NN	O	O
.	NN	O	O

mNFATc	NN	O	B-protein
complexed	NN	O	O
with	NN	O	O
AP-1	NN	O	B-protein
bound	NN	O	O
specifically	NN	O	O
to	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
IL-2	NN	O	I-protein
NFAT	NN	O	I-protein
recognition	NN	O	I-protein
sequence	NN	O	I-protein
and	NN	O	O
activated	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
co-transfected	NN	O	B-DNA
IL-2	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
COS-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
cDNA	NN	O	B-DNA
probe	NN	O	I-DNA
hybridized	NN	O	O
with	NN	O	O
a	NN	O	O
4.5	NN	O	O
kb	NN	O	O
transcript	NN	O	O
which	NN	O	O
is	NN	O	O
highly	NN	O	O
inducible	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

By	NN	O	O
Northern	NN	O	O
and	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
,	NN	O	O
mNFATc	NN	O	B-RNA
transcript	NN	O	I-RNA
was	NN	O	O
detected	NN	O	O
from	NN	O	O
the	NN	O	O
early	NN	O	O
stage	NN	O	O
of	NN	O	O
development	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
mouse	NN	O	O
embryo	NN	O	O
,	NN	O	O
mNFATc	NN	O	B-RNA
transcript	NN	O	I-RNA
was	NN	O	O
strongly	NN	O	O
expressed	NN	O	O
in	NN	O	O
thymus	NN	O	O
,	NN	O	O
lung	NN	O	O
and	NN	O	O
submandibular	NN	O	O
gland	NN	O	O
and	NN	O	O
weakly	NN	O	O
in	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
and	NN	O	O
heart	NN	O	O
suggesting	NN	O	O
that	NN	O	O
mNFATc	NN	O	B-protein
may	NN	O	O
have	NN	O	O
a	NN	O	O
role	NN	O	O
both	NN	O	O
in	NN	O	O
embryogenesis	NN	O	O
and	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

TNFalpha	NN	O	B-protein
cooperates	NN	O	O
with	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
pathway	NN	O	O
to	NN	O	O
synergistically	NN	O	O
increase	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
transcription	NN	O	O
via	NN	O	O
downstream	NN	O	O
TRE-like	NN	O	B-DNA
cAMP	NN	O	I-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Activating	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
binding	NN	O	O
TPA	NN	O	B-DNA
responsive	NN	O	I-DNA
elements	NN	O	I-DNA
(	NN	O	O
TRE	NN	O	B-DNA
)	NN	O	O
are	NN	O	O
located	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
U5	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

These	NN	O	O
downstream	NN	O	B-DNA
sequence	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
termed	NN	O	O
DSE	NN	O	B-DNA
,	NN	O	O
can	NN	O	O
bind	NN	O	O
both	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
CREB/ATF	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
DSE	NN	O	B-DNA
are	NN	O	O
also	NN	O	O
cAMP-responsive	NN	O	B-DNA
elements	NN	O	I-DNA
(	NN	O	O
CRE	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
since	NN	O	O
they	NN	O	O
mediated	NN	O	O
activation	NN	O	O
signals	NN	O	O
elicited	NN	O	O
by	NN	O	O
cholera	NN	O	B-protein
toxin	NN	O	I-protein
(	NN	O	O
Ctx	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
activator	NN	O	O
of	NN	O	O
the	NN	O	O
cAMP-dependent	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	O
PKA	NN	O	B-protein
)	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
DSE	NN	O	I-DNA
can	NN	O	O
mediate	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
synergy	NN	O	O
elicited	NN	O	O
by	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
Ctx	NN	O	B-protein
and	NN	O	O
TNFalpha	NN	O	B-protein
.	NN	O	O

Ctx	NN	O	B-protein
combined	NN	O	O
with	NN	O	O
TNFalpha	NN	O	B-protein
or	NN	O	O
IL-1beta	NN	O	B-protein
to	NN	O	O
produce	NN	O	O
a	NN	O	O
synergistic	NN	O	O
increase	NN	O	O
in	NN	O	O
p24	NN	O	B-protein
antigen	NN	O	I-protein
production	NN	O	O
in	NN	O	O
U1	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Transfection	NN	O	O
studies	NN	O	O
of	NN	O	O
LTR	NN	O	B-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
indicated	NN	O	O
that	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
DSE	NN	O	B-DNA
sites	NN	O	I-DNA
abrogated	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
-mediated	NN	O	O
synergy	NN	O	O
induced	NN	O	O
by	NN	O	O
Ctx	NN	O	B-protein
and	NN	O	O
TNFalpha	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
synergy	NN	O	O
induced	NN	O	O
by	NN	O	O
Ctx	NN	O	B-protein
and	NN	O	O
IL-1beta	NN	O	B-protein
was	NN	O	O
unaffected	NN	O	O
,	NN	O	O
suggesting	NN	O	O
TNFalpha	NN	O	B-protein
and	NN	O	O
IL-1beta	NN	O	B-protein
cooperate	NN	O	O
differently	NN	O	O
with	NN	O	O
the	NN	O	O
cAMP	NN	O	O
/PKA	NN	O	B-protein
activation	NN	O	O
pathway	NN	O	O
to	NN	O	O
induce	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
in	NN	O	O
U1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Because	NN	O	O
the	NN	O	O
DSE	NN	O	B-DNA
are	NN	O	O
also	NN	O	O
TRE	NN	O	B-DNA
sites	NN	O	O
,	NN	O	O
we	NN	O	O
assessed	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
agonist	NN	O	O
combinations	NN	O	O
on	NN	O	O
AP-1	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

TNFalpha	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
IL-1beta	NN	O	B-protein
cooperated	NN	O	O
with	NN	O	O
Ctx	NN	O	B-protein
to	NN	O	O
produce	NN	O	O
a	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
TRE-like	NN	O	B-DNA
cAMP-responsive	NN	O	I-DNA
DSE	NN	O	I-DNA
sites	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
5'-untranslated	NN	O	B-DNA
leader	NN	O	I-DNA
can	NN	O	O
mediate	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
cooperativity	NN	O	O
between	NN	O	O
TNFalpha	NN	O	B-protein
and	NN	O	O
the	NN	O	O
cAMP	NN	O	O
/PKA	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
DSE	NN	O	B-DNA
and	NN	O	O
TRE	NN	O	B-DNA
sites	NN	O	O
can	NN	O	O
not	NN	O	O
bind	NN	O	O
CREB/ATF	NN	O	B-protein
homodimers	NN	O	I-protein
,	NN	O	O
we	NN	O	O
propose	NN	O	O
a	NN	O	O
mechanism	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
DSE	NN	O	B-DNA
bind	NN	O	O
heterodimers	NN	O	B-protein
composed	NN	O	O
of	NN	O	O
both	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	I-protein
CREB/ATF	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Copyright	NN	O	O
1997	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Association	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
insensitivity	NN	O	O
with	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
beta	NN	O	I-protein
.	NN	O	O

In	NN	O	O
many	NN	O	O
chronic	NN	O	O
inflammatory	NN	O	O
disorders	NN	O	O
,	NN	O	O
glucocorticoid	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
insensitivity	NN	O	O
is	NN	O	O
a	NN	O	O
challenging	NN	O	O
clinical	NN	O	O
problem	NN	O	O
associated	NN	O	O
with	NN	O	O
life-threatening	NN	O	O
disease	NN	O	O
progression	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
GC	NN	O	O
insensitivity	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

Alternative	NN	O	O
splicing	NN	O	O
of	NN	O	O
the	NN	O	O
GC	NN	O	B-RNA
receptor	NN	O	I-RNA
(	NN	O	I-RNA
R	NN	O	I-RNA
)	NN	O	I-RNA
pre-messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
generates	NN	O	O
a	NN	O	O
second	NN	O	O
GCR	NN	O	B-protein
,	NN	O	O
termed	NN	O	O
GCR-beta	NN	O	B-protein
,	NN	O	O
which	NN	O	O
does	NN	O	O
not	NN	O	O
bind	NN	O	O
GCs	NN	O	O
but	NN	O	O
antagonizes	NN	O	O
the	NN	O	O
transactivating	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
classic	NN	O	O
GCR	NN	O	B-protein
,	NN	O	O
termed	NN	O	O
GCR-alpha	NN	O	B-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
current	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
GC-insensitive	NN	O	O
asthma	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
significantly	NN	O	O
higher	NN	O	O
number	NN	O	O
of	NN	O	O
GCR-beta-immunoreactive	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
than	NN	O	O
GC-sensitive	NN	O	O
asthmatics	NN	O	O
or	NN	O	O
normal	NN	O	O
controls	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
patients	NN	O	O
with	NN	O	O
GC-insensitive	NN	O	O
asthma	NN	O	O
have	NN	O	O
cytokine-induced	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
capability	NN	O	O
of	NN	O	O
the	NN	O	O
GCR	NN	O	B-protein
.	NN	O	O

These	NN	O	O
abnormalities	NN	O	O
can	NN	O	O
be	NN	O	O
reproduced	NN	O	O
by	NN	O	O
transfection	NN	O	O
of	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
GCR-beta	NN	O	B-protein
gene	NN	O	O
resulting	NN	O	O
in	NN	O	O
significant	NN	O	O
reduction	NN	O	O
of	NN	O	O
their	NN	O	O
GCR-alpha	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
capacity	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
GCR-beta	NN	O	B-protein
is	NN	O	O
cytokine	NN	O	B-protein
inducible	NN	O	O
and	NN	O	O
may	NN	O	O
account	NN	O	O
for	NN	O	O
GC	NN	O	O
insensitivity	NN	O	O
in	NN	O	O
this	NN	O	O
common	NN	O	O
inflammatory	NN	O	O
condition	NN	O	O
.	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	O
virus	NN	O	O
EBNA3C	NN	O	B-protein
represses	NN	O	O
Cp	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
major	NN	O	O
promoter	NN	O	O
for	NN	O	O
EBNA	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
but	NN	O	O
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
promoter	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	B-DNA
gene	NN	O	I-DNA
CD21	NN	O	I-DNA
.	NN	O	O

EBNA3C	NN	O	B-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
repressor	NN	O	O
of	NN	O	O
transcription	NN	O	O
when	NN	O	O
bound	NN	O	O
to	NN	O	O
DNA	NN	O	O
as	NN	O	O
a	NN	O	O
fusion	NN	O	O
with	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
(	NN	O	O
DBD	NN	O	B-protein
)	NN	O	O
of	NN	O	O
GALA	NN	O	O
.	NN	O	O

A	NN	O	O
survey	NN	O	O
of	NN	O	O
promoters	NN	O	O
has	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
,	NN	O	I-protein
unfused	NN	O	I-protein
EBNA3C	NN	O	I-protein
can	NN	O	O
specifically	NN	O	O
repress	NN	O	O
expression	NN	O	O
from	NN	O	O
reporter	NN	O	B-DNA
plasmids	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
Cp	NN	O	B-DNA
latency-associated	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Repression	NN	O	O
of	NN	O	O
Cp	NN	O	B-DNA
activity	NN	O	O
required	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
207	NN	O	I-protein
to	NN	O	I-protein
368	NN	O	I-protein
,	NN	O	O
which	NN	O	O
encompasses	NN	O	O
a	NN	O	O
region	NN	O	O
resembling	NN	O	O
a	NN	O	O
basic	NN	O	B-protein
DBD	NN	O	I-protein
adjacent	NN	O	O
to	NN	O	O
a	NN	O	O
leucine	NN	O	B-protein
zipper	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
motif	NN	O	I-protein
and	NN	O	O
a	NN	O	O
site	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
factor	NN	O	I-protein
CBF1/RBP-Jkappa	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
amino	NN	O	O
acids	NN	O	O
207	NN	O	O
to	NN	O	O
368	NN	O	O
are	NN	O	O
dispensable	NN	O	O
when	NN	O	O
the	NN	O	O
protein	NN	O	O
is	NN	O	O
bound	NN	O	O
to	NN	O	O
DNA	NN	O	O
as	NN	O	O
a	NN	O	O
fusion	NN	O	O
with	NN	O	O
the	NN	O	O
GAL4	NN	O	O
DBD	NN	O	B-protein
,	NN	O	O
thus	NN	O	O
implicating	NN	O	O
this	NN	O	O
region	NN	O	O
in	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
CBF1/RBP-Jkappa	NN	O	B-protein
binding	NN	O	I-protein
site	NN	O	I-protein
in	NN	O	O
EBNA3C	NN	O	B-protein
abrogated	NN	O	O
repression	NN	O	O
,	NN	O	O
strongly	NN	O	O
suggesting	NN	O	O
that	NN	O	O
CBF1/RBP-Jkappa	NN	O	B-protein
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
targeting	NN	O	O
the	NN	O	O
viral	NN	O	O
protein	NN	O	O
to	NN	O	O
Cp	NN	O	B-DNA
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
this	NN	O	O
result	NN	O	O
,	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
EBNA2	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
a	NN	O	O
CBF1/RBP-Jkappa	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
Cp	NN	O	B-DNA
also	NN	O	O
prevented	NN	O	O
significant	NN	O	O
repression	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
346	NN	O	I-protein
to	NN	O	I-protein
543	NN	O	I-protein
,	NN	O	O
which	NN	O	O
were	NN	O	O
previously	NN	O	O
defined	NN	O	O
as	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
repressor	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
GAL4-EBNA3C	NN	O	B-protein
fusion	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
also	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
repression	NN	O	O
of	NN	O	O
Cp	NN	O	B-DNA
.	NN	O	O

Since	NN	O	O
repression	NN	O	O
by	NN	O	O
these	NN	O	O
fusions	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
all	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
it	NN	O	O
seems	NN	O	O
likely	NN	O	O
that	NN	O	O
EBNA3C	NN	O	B-protein
either	NN	O	O
depends	NN	O	O
on	NN	O	O
a	NN	O	O
corepressor	NN	O	B-protein
which	NN	O	O
may	NN	O	O
interact	NN	O	O
with	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
346	NN	O	I-protein
to	NN	O	I-protein
543	NN	O	I-protein
or	NN	O	O
is	NN	O	O
modified	NN	O	O
in	NN	O	O
a	NN	O	O
cell-specific	NN	O	O
manner	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
repress	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
EBNA3C	NN	O	B-protein
contributing	NN	O	O
to	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
EBNA	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
latently	NN	O	B-cell_type
infected	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
CBF1/RBP-Jkappa	NN	O	B-protein
and	NN	O	O
another	NN	O	O
factor	NN	O	O
,	NN	O	O
but	NN	O	O
this	NN	O	O
need	NN	O	O
not	NN	O	O
directly	NN	O	O
involve	NN	O	O
EBNA2	NN	O	B-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
although	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
that	NN	O	O
EBNA3C	NN	O	B-protein
can	NN	O	O
upregulate	NN	O	O
CD21	NN	O	B-protein
in	NN	O	O
some	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
any	NN	O	O
effect	NN	O	O
of	NN	O	O
EBNA3C	NN	O	B-protein
on	NN	O	O
reporter	NN	O	B-DNA
plasmids	NN	O	I-DNA
which	NN	O	O
contain	NN	O	O
the	NN	O	O
CD21	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Relationship	NN	O	O
between	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
response	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
in	NN	O	O
ulcerative	NN	O	O
colitis	NN	O	O
.	NN	O	O

PURPOSE	NN	O	O
:	NN	O	O
To	NN	O	O
clarify	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
the	NN	O	O
effectiveness	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
ulcerative	NN	O	O
colitis	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
number	NN	O	O
and	NN	O	O
apparent	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
ulcerative	NN	O	O
colitis	NN	O	O
.	NN	O	O

MATERIALS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
Eleven	NN	O	O
patients	NN	O	O
with	NN	O	O
ulcerative	NN	O	O
colitis	NN	O	O
(	NN	O	O
5	NN	O	O
who	NN	O	O
responded	NN	O	O
to	NN	O	O
intravenous	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
6	NN	O	O
who	NN	O	O
did	NN	O	O
not	NN	O	O
)	NN	O	O
and	NN	O	O
ten	NN	O	O
control	NN	O	O
subjects	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
and	NN	O	O
apparent	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
were	NN	O	O
measured	NN	O	O
using	NN	O	O
a	NN	O	O
whole-cell	NN	O	O
binding	NN	O	O
assay	NN	O	O
.	NN	O	O

Results	NN	O	O
were	NN	O	O
expressed	NN	O	O
as	NN	O	O
a	NN	O	O
median	NN	O	O
(	NN	O	O
interquartile	NN	O	O
range	NN	O	O
)	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
The	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
from	NN	O	O
the	NN	O	O
six	NN	O	O
nonresponders	NN	O	O
,	NN	O	O
five	NN	O	O
responders	NN	O	O
,	NN	O	O
and	NN	O	O
ten	NN	O	O
healthy	NN	O	O
controls	NN	O	O
were	NN	O	O
4922	NN	O	O
(	NN	O	O
range	NN	O	O
,	NN	O	O
4484-5643	NN	O	O
)	NN	O	O
,	NN	O	O
3413	NN	O	O
(	NN	O	O
range	NN	O	O
,	NN	O	O
3183-4450	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
3610	NN	O	O
(	NN	O	O
range	NN	O	O
,	NN	O	O
2594-3979	NN	O	O
)	NN	O	O
binding	NN	O	O
sites/cell	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
apparent	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
from	NN	O	O
the	NN	O	O
nonresponders	NN	O	O
,	NN	O	O
responders	NN	O	O
,	NN	O	O
and	NN	O	O
healthy	NN	O	O
controls	NN	O	O
were	NN	O	O
7.03	NN	O	O
(	NN	O	O
range	NN	O	O
,	NN	O	O
5.66-10	NN	O	O
)	NN	O	O
,	NN	O	O
4.27	NN	O	O
(	NN	O	O
range	NN	O	O
,	NN	O	O
4-5.13	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
6.18	NN	O	O
(	NN	O	O
range	NN	O	O
,	NN	O	O
5.86-6.74	NN	O	O
)	NN	O	O
nM	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Nonresponders	NN	O	O
had	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
both	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
binding	NN	O	B-protein
sites	NN	O	I-protein
and	NN	O	O
in	NN	O	O
the	NN	O	O
apparent	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
compared	NN	O	O
with	NN	O	O
responders	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.045	NN	O	O
;	NN	O	O
P	NN	O	O
=	NN	O	O
0.029	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
The	NN	O	O
increased	NN	O	O
number	NN	O	O
and	NN	O	O
apparent	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
are	NN	O	O
closely	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
effectiveness	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
.	NN	O	O

The	NN	O	O
measurement	NN	O	O
of	NN	O	O
the	NN	O	O
number	NN	O	O
and	NN	O	O
apparent	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
may	NN	O	O
be	NN	O	O
useful	NN	O	O
in	NN	O	O
predicting	NN	O	O
response	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cooperation	NN	O	O
of	NN	O	O
binding	NN	O	B-protein
sites	NN	O	I-protein
for	NN	O	O
STAT6	NN	O	B-protein
and	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B/rel	NN	O	I-protein
in	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IgE	NN	O	I-DNA
germline	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Ig	NN	O	B-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
class	NN	O	O
switching	NN	O	O
is	NN	O	O
directed	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
inducing	NN	O	O
transcription	NN	O	O
from	NN	O	O
unrearranged	NN	O	O
CH	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Subsequently	NN	O	O
,	NN	O	O
such	NN	O	O
primed	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
undergo	NN	O	O
switch	NN	O	O
recombination	NN	O	O
to	NN	O	O
express	NN	O	O
the	NN	O	O
selected	NN	O	O
new	NN	O	O
isotype	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
class	NN	O	O
switching	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
activates	NN	O	O
the	NN	O	O
IgE	NN	O	B-DNA
germline	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
inducing	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
STAT6	NN	O	B-protein
(	NN	O	O
IL-4STAT	NN	O	B-protein
)	NN	O	O
with	NN	O	O
a	NN	O	O
responsive	NN	O	B-DNA
DNA	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

This	NN	O	O
study	NN	O	O
describes	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
two	NN	O	O
additional	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
that	NN	O	O
interact	NN	O	O
with	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B/rel	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
in	NN	O	O
an	NN	O	O
IL-4	NN	O	B-protein
-independent	NN	O	O
fashion	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
nucleoprotein	NN	O	B-protein
complex	NN	O	I-protein
formed	NN	O	O
on	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
site	NN	O	I-DNA
(	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B1	NN	O	I-protein
)	NN	O	O
contains	NN	O	O
the	NN	O	O
classical	NN	O	B-protein
p50/p65	NN	O	I-protein
heterodimer	NN	O	I-protein
.	NN	O	O

The	NN	O	O
complex	NN	O	O
on	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
site	NN	O	I-DNA
(	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B2	NN	O	I-protein
)	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
composed	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
relB	NN	O	B-protein
.	NN	O	O

IgE	NN	O	B-DNA
germline	NN	O	I-DNA
promoter	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
carrying	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B2	NN	O	I-protein
site	NN	O	O
were	NN	O	O
largely	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
stimulation	NN	O	O
in	NN	O	O
transient	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
,	NN	O	O
while	NN	O	O
plasmids	NN	O	O
with	NN	O	O
similar	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B1	NN	O	I-protein
site	NN	O	O
responded	NN	O	O
to	NN	O	O
cytokine	NN	O	B-protein
stimulation	NN	O	O
better	NN	O	O
than	NN	O	O
the	NN	O	O
wild-type	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B2	NN	O	I-protein
effect	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
STAT6	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B2	NN	O	I-protein
motif	NN	O	I-protein
is	NN	O	O
necessary	NN	O	O
but	NN	O	O
not	NN	O	O
sufficient	NN	O	O
for	NN	O	O
mediating	NN	O	O
cytokine	NN	O	B-protein
up-regulation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
a	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B/rel	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
STAT6	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
conferred	NN	O	O
IL-4	NN	O	B-protein
inducibility	NN	O	O
to	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
minimal	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
while	NN	O	O
the	NN	O	O
individual	NN	O	O
sites	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

The	NN	O	O
available	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B2	NN	O	I-protein
nucleoprotein	NN	O	I-protein
complex	NN	O	I-protein
may	NN	O	O
cooperate	NN	O	O
with	NN	O	O
DNA-bound	NN	O	B-protein
STAT6	NN	O	I-protein
to	NN	O	O
achieve	NN	O	O
IL-4	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IgE	NN	O	I-DNA
germline	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Alcohol-induced	NN	O	O
regulation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
regulatory	NN	O	I-protein
factor-kappa	NN	O	I-protein
beta	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Acute	NN	O	O
ethanol	NN	O	O
exposure	NN	O	O
has	NN	O	O
the	NN	O	O
capacity	NN	O	O
to	NN	O	O
modulate	NN	O	O
immune	NN	O	O
functions	NN	O	O
,	NN	O	O
particularly	NN	O	O
,	NN	O	O
to	NN	O	O
down	NN	O	O
regulate	NN	O	O
monocyte	NN	O	O
production	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
intracellular	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
these	NN	O	O
effects	NN	O	O
of	NN	O	O
ethanol	NN	O	O
are	NN	O	O
yet	NN	O	O
to	NN	O	O
be	NN	O	O
understood	NN	O	O
.	NN	O	O

Considering	NN	O	O
that	NN	O	O
nuclear	NN	O	B-protein
regulatory	NN	O	I-protein
factor-kappa	NN	O	I-protein
beta	NN	O	I-protein
(	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
/Rel	NN	O	I-protein
is	NN	O	O
a	NN	O	O
common	NN	O	O
regulatory	NN	O	B-protein
element	NN	O	I-protein
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
inflammatory	NN	O	B-DNA
cytokine	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
herein	NN	O	O
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
acute	NN	O	O
ethanol	NN	O	O
affects	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Adherence-isolated	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
showed	NN	O	O
constitutive	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

A	NN	O	O
clinically	NN	O	O
relevant	NN	O	O
dose	NN	O	O
(	NN	O	O
25	NN	O	O
mM	NN	O	O
)	NN	O	O
of	NN	O	O
acute	NN	O	O
ethanol	NN	O	O
treatment	NN	O	O
in	NN	O	O
vitro	NN	O	O
increased	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
with	NN	O	O
a	NN	O	O
preferential	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
,	NN	O	O
p50/p50	NN	O	B-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
homodimer	NN	O	I-protein
,	NN	O	O
and	NN	O	O
resulted	NN	O	O
in	NN	O	O
no	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
p65/p50	NN	O	B-protein
heterodimer	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
lipopolysaccharide	NN	O	O
stimulation	NN	O	O
primarily	NN	O	O
induced	NN	O	O
the	NN	O	O
p65/p50	NN	O	B-protein
heterodimer	NN	O	I-protein
that	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
gene	NN	O	O
activation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
such	NN	O	O
unique	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	B-protein
p50/p50	NN	O	I-protein
homodimer	NN	O	I-protein
by	NN	O	O
acute	NN	O	O
ethanol	NN	O	O
treatment	NN	O	O
may	NN	O	O
result	NN	O	O
in	NN	O	O
inhibition	NN	O	O
rather	NN	O	O
than	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B-regulated	NN	O	I-DNA
inflammatory	NN	O	I-DNA
cytokine	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
physiologically	NN	O	O
relevant	NN	O	O
concentrations	NN	O	O
of	NN	O	O
ethanol	NN	O	O
may	NN	O	O
affect	NN	O	O
production	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
,	NN	O	O
interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
and	NN	O	O
interleukin-6	NN	O	B-protein
by	NN	O	O
disrupting	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
signaling	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

LPS	NN	O	O
tolerance	NN	O	O
in	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
:	NN	O	O
three	NN	O	O
3	NN	O	O
'	NN	O	O
cytosins	NN	O	O
are	NN	O	O
required	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
for	NN	O	O
detection	NN	O	O
of	NN	O	O
upregulated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	I-protein
homodimers	NN	O	I-protein
.	NN	O	O

When	NN	O	O
monocytes	NN	O	B-cell_type
are	NN	O	O
stimulated	NN	O	O
with	NN	O	O
LPS	NN	O	O
(	NN	O	O
lipopolysaccharide	NN	O	O
)	NN	O	O
repeatedly	NN	O	O
then	NN	O	O
the	NN	O	O
initially	NN	O	O
high	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
(	NN	O	I-DNA
tumor	NN	O	I-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
only	NN	O	O
very	NN	O	O
low	NN	O	O
,	NN	O	O
i.e.	NN	O	O
the	NN	O	O
cells	NN	O	O
are	NN	O	O
tolerant	NN	O	O
to	NN	O	O
LPS	NN	O	O
.	NN	O	O

Tolerant	NN	O	O
cells	NN	O	O
still	NN	O	O
express	NN	O	O
the	NN	O	O
CD14	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
they	NN	O	O
can	NN	O	O
still	NN	O	O
be	NN	O	O
activated	NN	O	O
to	NN	O	O
mobilize	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
into	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	O
proteins	NN	O	O
employing	NN	O	O
the	NN	O	O
-605	NN	O	O
motif	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
TNF	NN	O	I-DNA
promoter	NN	O	I-DNA
(	NN	O	O
GGGGCTGTCCC	NN	O	O
)	NN	O	O
revealed	NN	O	O
that	NN	O	O
in	NN	O	O
tolerant	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
there	NN	O	O
is	NN	O	O
a	NN	O	O
predominance	NN	O	O
of	NN	O	O
p50p50	NN	O	B-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
mutant	NN	O	B-DNA
motif	NN	O	I-DNA
that	NN	O	O
exchanges	NN	O	O
the	NN	O	O
terminal	NN	O	O
3	NN	O	O
'	NN	O	O
C	NN	O	O
for	NN	O	O
a	NN	O	O
G	NN	O	O
fails	NN	O	O
to	NN	O	O
bind	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
homodimer	NN	O	I-protein
that	NN	O	O
is	NN	O	O
upregulated	NN	O	O
in	NN	O	O
LPS	NN	O	O
toler	NN	O	O
ant	NN	O	O
human	NN	O	B-cell_line
Mono	NN	O	I-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
same	NN	O	O
is	NN	O	O
true	NN	O	O
for	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
taken	NN	O	O
from	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
P388D1	NN	O	I-cell_line
macrophage	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
when	NN	O	O
tested	NN	O	O
with	NN	O	O
the	NN	O	O
-516	NN	O	B-DNA
motif	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
TNF	NN	O	I-DNA
promoter	NN	O	I-DNA
(	NN	O	O
GGGGGCTTTCCC	NN	O	O
)	NN	O	O
.	NN	O	O

Here	NN	O	O
the	NN	O	O
wild	NN	O	B-DNA
type	NN	O	I-DNA
motif	NN	O	I-DNA
gives	NN	O	O
efficient	NN	O	O
binding	NN	O	O
of	NN	O	O
p50p50	NN	O	B-protein
that	NN	O	O
again	NN	O	O
is	NN	O	O
upregulated	NN	O	O
in	NN	O	O
tolerant	NN	O	B-cell_type
cells	NN	O	I-cell_type
whereas	NN	O	O
a	NN	O	O
mutant	NN	O	O
with	NN	O	O
a	NN	O	O
3	NN	O	O
'	NN	O	O
G	NN	O	O
shows	NN	O	O
hardly	NN	O	O
any	NN	O	O
binding	NN	O	O
of	NN	O	O
p50p50	NN	O	B-protein
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
kappa	NN	O	I-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
motif	NN	O	I-DNA
(	NN	O	O
GGGGACTTTCCG	NN	O	O
)	NN	O	O
does	NN	O	O
not	NN	O	O
efficiently	NN	O	O
bind	NN	O	O
the	NN	O	O
nuclear	NN	O	O
p50p50	NN	O	B-protein
from	NN	O	O
tolerant	NN	O	B-cell_line
murine	NN	O	I-cell_line
P388	NN	O	I-cell_line
macrophages	NN	O	I-cell_line
.	NN	O	O

Binding	NN	O	O
is	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
readily	NN	O	O
detected	NN	O	O
when	NN	O	O
the	NN	O	O
3	NN	O	O
'	NN	O	O
G	NN	O	O
is	NN	O	O
replaced	NN	O	O
by	NN	O	O
a	NN	O	O
C	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
upregulated	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	I-protein
in	NN	O	O
LPS	NN	O	O
tolerant	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
subtle	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Cyclosporin	NN	O	O
A	NN	O	O
inhibits	NN	O	O
early	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
G0/G1	NN	O	B-DNA
switch	NN	O	I-DNA
gene	NN	O	I-DNA
2	NN	O	I-DNA
(	NN	O	O
G0S2	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
blood	NN	O	I-cell_line
mononuclear	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
may	NN	O	O
achieve	NN	O	O
its	NN	O	O
immunosuppressive	NN	O	O
effects	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
calcium-	NN	O	B-protein
and	NN	O	I-protein
calmodulin-dependent	NN	O	I-protein
phosphatase	NN	O	I-protein
calcineurin	NN	O	B-protein
which	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
by	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
(	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
)	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Among	NN	O	O
these	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
facilitating	NN	O	O
progression	NN	O	O
through	NN	O	O
the	NN	O	O
G1	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
IL2	NN	O	B-protein
does	NN	O	O
not	NN	O	O
reverse	NN	O	O
CsA	NN	O	O
inhibition	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
other	NN	O	O
NFAT-sensitive	NN	O	B-DNA
gene	NN	O	I-DNA
may	NN	O	O
be	NN	O	O
involved	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
G0/G1	NN	O	I-DNA
switch	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
G0S2	NN	O	B-DNA
,	NN	O	O
has	NN	O	O
potential	NN	O	O
NFAT-binding	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flank	NN	O	I-DNA
and	NN	O	O
encodes	NN	O	O
a	NN	O	O
small	NN	O	B-protein
basic	NN	O	I-protein
potential	NN	O	I-protein
phosphoprotein	NN	O	I-protein
of	NN	O	O
unknown	NN	O	O
function	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
sensitive	NN	O	O
,	NN	O	O
reverse	NN	O	O
transcription-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
assay	NN	O	O
,	NN	O	O
G0S2	NN	O	B-DNA
mRNA	NN	O	O
levels	NN	O	O
were	NN	O	O
assayed	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_line
blood	NN	O	I-cell_line
mononuclear	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Freshly	NN	O	O
isolated	NN	O	O
cells	NN	O	O
contain	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
G0S2	NN	O	B-RNA
mRNA	NN	O	I-RNA
which	NN	O	O
rapidly	NN	O	O
decline	NN	O	O
.	NN	O	O

This	NN	O	O
``	NN	O	O
spontaneous	NN	O	O
stimulation	NN	O	O
''	NN	O	O
is	NN	O	O
also	NN	O	O
noted	NN	O	O
with	NN	O	O
some	NN	O	O
other	NN	O	O
G0S	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
has	NN	O	O
been	NN	O	O
attributed	NN	O	O
to	NN	O	O
some	NN	O	O
aspect	NN	O	O
of	NN	O	O
the	NN	O	O
isolation	NN	O	O
procedure	NN	O	O
.	NN	O	O

In	NN	O	O
cells	NN	O	O
that	NN	O	O
have	NN	O	O
been	NN	O	O
preincubated	NN	O	O
to	NN	O	O
lower	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
transient	NN	O	O
increase	NN	O	O
in	NN	O	O
G0S2	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
peaking	NN	O	O
between	NN	O	O
1-2	NN	O	O
h	NN	O	O
,	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
Concanavalin-A	NN	O	O
(	NN	O	O
ConA	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
to	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
,	NN	O	O
ionomycin	NN	O	O
.	NN	O	O

Both	NN	O	O
these	NN	O	O
responses	NN	O	O
are	NN	O	O
inhibited	NN	O	O
by	NN	O	O
CsA	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
G0S2	NN	O	B-DNA
expression	NN	O	O
is	NN	O	O
required	NN	O	O
to	NN	O	O
commit	NN	O	O
cells	NN	O	O
to	NN	O	O
enter	NN	O	O
the	NN	O	O
G1	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
,	NN	O	O
while	NN	O	O
not	NN	O	O
excluding	NN	O	O
other	NN	O	O
possible	NN	O	O
targets	NN	O	O
,	NN	O	O
early	NN	O	O
inhibition	NN	O	O
of	NN	O	O
G0S2	NN	O	B-DNA
expression	NN	O	O
by	NN	O	O
CsA	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
achieving	NN	O	O
immunosuppression	NN	O	O
.	NN	O	O

G0S2	NN	O	B-DNA
may	NN	O	O
be	NN	O	O
of	NN	O	O
value	NN	O	O
as	NN	O	O
a	NN	O	O
reporter	NN	O	O
gene	NN	O	O
for	NN	O	O
analyzing	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
CsA	NN	O	O
and	NN	O	O
its	NN	O	O
influence	NN	O	O
on	NN	O	O
the	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	O
selection	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
self	NN	O	O
and	NN	O	O
not-self	NN	O	O
antigens	NN	O	O
.	NN	O	O

-DOCSTART-	O

c-Rel	NN	O	B-protein
and	NN	O	I-protein
p65	NN	O	I-protein
subunits	NN	O	I-protein
bind	NN	O	O
to	NN	O	O
an	NN	O	O
upstream	NN	O	B-DNA
NF-kappaB	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-DNA
granulocyte	NN	O	I-DNA
macrophage-colony	NN	O	I-DNA
stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
promoter	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
response	NN	O	O
in	NN	O	O
5637	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
further	NN	O	O
clarify	NN	O	O
the	NN	O	O
complex	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
was	NN	O	O
extensively	NN	O	O
investigated	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
an	NN	O	O
upstream	NN	O	O
NF-kappaB	NN	O	B-DNA
like	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
5637	NN	O	B-cell_line
non-lymphoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
derives	NN	O	O
from	NN	O	O
a	NN	O	O
bladder	NN	O	O
carcinoma	NN	O	O
and	NN	O	O
constitutively	NN	O	O
produces	NN	O	O
GM-CSF	NN	O	B-protein
.	NN	O	O

This	NN	O	O
sequence	NN	O	O
,	NN	O	O
named	NN	O	O
the	NN	O	O
A	NN	O	O
element	NN	O	O
,	NN	O	O
has	NN	O	O
an	NN	O	O
active	NN	O	O
role	NN	O	O
on	NN	O	O
GM-CSF	NN	O	B-protein
transcription	NN	O	O
and	NN	O	O
is	NN	O	O
responsive	NN	O	O
to	NN	O	O
the	NN	O	O
tumor	NN	O	B-DNA
promoter	NN	O	I-DNA
PMA	NN	O	I-DNA
in	NN	O	O
transient	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
here	NN	O	O
a	NN	O	O
heterodimeric	NN	O	B-protein
binding	NN	O	I-protein
complex	NN	O	I-protein
of	NN	O	O
NF-kappaB	NN	O	B-protein
subunits	NN	O	I-protein
(	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
)	NN	O	O
which	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
one	NN	O	O
obtained	NN	O	O
using	NN	O	O
the	NN	O	O
HIV-LTR-kappaB	NN	O	B-DNA
site	NN	O	I-DNA
as	NN	O	O
recognition	NN	O	O
sequence	NN	O	O
and	NN	O	O
different	NN	O	O
from	NN	O	O
the	NN	O	O
one	NN	O	O
(	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	O
p50	NN	O	B-protein
)	NN	O	O
observed	NN	O	O
with	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
Mo	NN	O	B-cell_line
T-lymphoid	NN	O	I-cell_line
HTLV-II	NN	O	I-cell_line
infected	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Repression	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
through	NN	O	O
the	NN	O	O
novel	NN	O	O
cooperation	NN	O	O
of	NN	O	O
human	NN	O	B-protein
factors	NN	O	I-protein
YY1	NN	O	B-protein
and	NN	O	O
LSF	NN	O	B-protein
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
J	NN	O	O
Virol	NN	O	O
1998	NN	O	O
Feb	NN	O	O
;	NN	O	O
72	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
:	NN	O	O
1709	NN	O	O
]	NN	O	O

A	NN	O	O
subpopulation	NN	O	O
of	NN	O	O
stably	NN	O	B-cell_type
infected	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
cells	NN	O	I-cell_type
capable	NN	O	O
of	NN	O	O
producing	NN	O	O
virus	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
-positive	NN	O	O
individuals	NN	O	O
(	NN	O	O
T.-W.Chun	NN	O	O
,	NN	O	O
D.Finzi	NN	O	O
,	NN	O	O
J.Margolick	NN	O	O
,	NN	O	O
K.Chadwick	NN	O	O
,	NN	O	O
D.Schwartz	NN	O	O
,	NN	O	O
and	NN	O	O
R.F.Siliciano	NN	O	O
,	NN	O	O
Nat.Med.1	NN	O	O
:	NN	O	O
1284-1290	NN	O	O
,	NN	O	O
1995	NN	O	O
)	NN	O	O
.	NN	O	O

Few	NN	O	O
host	NN	O	O
factors	NN	O	O
that	NN	O	O
directly	NN	O	O
limit	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
and	NN	O	O
could	NN	O	O
support	NN	O	O
this	NN	O	O
state	NN	O	O
of	NN	O	O
nonproductive	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

YY1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
widely	NN	O	O
distributed	NN	O	O
human	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
inhibit	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
transcription	NN	O	O
and	NN	O	O
virus	NN	O	O
production	NN	O	O
.	NN	O	O

LSF	NN	O	B-protein
(	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
LBP-1	NN	O	B-protein
,	NN	O	O
UBP	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CP-2	NN	O	B-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
repress	NN	O	O
LTR	NN	O	O
transcription	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
but	NN	O	O
transient	NN	O	O
expression	NN	O	O
of	NN	O	O
LSF	NN	O	B-protein
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
LTR	NN	O	B-DNA
activity	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
both	NN	O	O
YY1	NN	O	B-protein
and	NN	O	O
LSF	NN	O	B-protein
participate	NN	O	O
in	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
complex	NN	O	O
that	NN	O	O
recognizes	NN	O	O
the	NN	O	O
initiation	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

Further	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
these	NN	O	O
factors	NN	O	O
cooperate	NN	O	O
in	NN	O	O
the	NN	O	O
repression	NN	O	O
of	NN	O	O
LTR	NN	O	B-DNA
expression	NN	O	O
and	NN	O	O
viral	NN	O	O
replication	NN	O	O
.	NN	O	O

This	NN	O	O
cooperative	NN	O	O
function	NN	O	O
may	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
divergent	NN	O	O
effects	NN	O	O
of	NN	O	O
LSF	NN	O	B-protein
previously	NN	O	O
observed	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
cooperation	NN	O	O
of	NN	O	O
two	NN	O	O
general	NN	O	O
cellular	NN	O	O
transcription	NN	O	O
factors	NN	O	O
may	NN	O	O
allow	NN	O	O
for	NN	O	O
the	NN	O	O
selective	NN	O	O
downregulation	NN	O	O
of	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
.	NN	O	O

Through	NN	O	O
this	NN	O	O
mechanism	NN	O	O
of	NN	O	O
gene	NN	O	O
regulation	NN	O	O
,	NN	O	O
YY1	NN	O	B-protein
and	NN	O	O
LSF	NN	O	B-protein
could	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
establishment	NN	O	O
and	NN	O	O
maintenance	NN	O	O
of	NN	O	O
a	NN	O	O
population	NN	O	O
of	NN	O	O
cells	NN	O	O
stably	NN	O	O
but	NN	O	O
nonproductively	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

-DOCSTART-	O

Analysis	NN	O	O
of	NN	O	O
interactions	NN	O	O
between	NN	O	O
huGATA-3	NN	O	B-protein
transcription	NN	O	O
factor	NN	O	O
and	NN	O	O
three	NN	O	O
GATA	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
of	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
,	NN	O	O
by	NN	O	O
surface	NN	O	O
plasmon	NN	O	O
resonance	NN	O	O
.	NN	O	O

Relative	NN	O	O
affinities	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
for	NN	O	O
their	NN	O	O
respective	NN	O	O
factor	NN	O	O
have	NN	O	O
been	NN	O	O
essentially	NN	O	O
studied	NN	O	O
by	NN	O	O
bandshift	NN	O	O
analysis	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
a	NN	O	O
real-time	NN	O	O
study	NN	O	O
of	NN	O	O
factor/DNA	NN	O	O
interactions	NN	O	O
using	NN	O	O
a	NN	O	O
surface	NN	O	O
plasmon	NN	O	O
resonance	NN	O	O
approach	NN	O	O
and	NN	O	O
further	NN	O	O
characterization	NN	O	O
of	NN	O	O
recovered	NN	O	B-protein
proteins	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
this	NN	O	O
interaction	NN	O	O
.	NN	O	O

For	NN	O	O
this	NN	O	O
purpose	NN	O	O
,	NN	O	O
human	NN	O	B-protein
GATA-3	NN	O	I-protein
,	NN	O	O
either	NN	O	O
recombinant	NN	O	O
or	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
,	NN	O	O
and	NN	O	O
three	NN	O	O
natural	NN	O	B-DNA
GATA	NN	O	I-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
sites	NN	O	B-DNA
1	NN	O	I-DNA
,	NN	O	I-DNA
2	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
3	NN	O	I-DNA
)	NN	O	O
were	NN	O	O
chosen	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
only	NN	O	O
site	NN	O	B-DNA
2	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
noncanonical	NN	O	O
GATA	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

Direct	NN	O	O
analysis	NN	O	O
of	NN	O	O
sensorgrams	NN	O	O
,	NN	O	O
with	NN	O	O
recombinant	NN	O	B-protein
huGATA-3	NN	O	I-protein
,	NN	O	O
allowed	NN	O	O
the	NN	O	O
comparison	NN	O	O
of	NN	O	O
association	NN	O	O
and	NN	O	O
dissociation	NN	O	O
profiles	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
DNA	NN	O	B-DNA
regions	NN	O	I-DNA
and	NN	O	O
their	NN	O	O
ranking	NN	O	O
according	NN	O	O
to	NN	O	O
their	NN	O	O
relative	NN	O	O
affinities	NN	O	O
.	NN	O	O

This	NN	O	O
result	NN	O	O
,	NN	O	O
confirmed	NN	O	O
by	NN	O	O
competitions	NN	O	O
with	NN	O	O
each	NN	O	O
GATA	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
higher	NN	O	O
relative	NN	O	O
affinity	NN	O	O
(	NN	O	O
at	NN	O	O
least	NN	O	O
sevenfold	NN	O	O
)	NN	O	O
of	NN	O	O
site	NN	O	O
3	NN	O	O
.	NN	O	O

Interactions	NN	O	O
between	NN	O	O
the	NN	O	O
canonical	NN	O	O
and	NN	O	O
unique	NN	O	O
GATA	NN	O	B-DNA
site	NN	O	I-DNA
3	NN	O	I-DNA
and	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
were	NN	O	O
also	NN	O	O
studied	NN	O	O
in	NN	O	O
real	NN	O	O
time	NN	O	O
and	NN	O	O
provided	NN	O	O
information	NN	O	O
on	NN	O	O
its	NN	O	O
association	NN	O	O
and	NN	O	O
dissociation	NN	O	O
rates	NN	O	O
for	NN	O	O
native	NN	O	O
huGATA-3	NN	O	B-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
recovered	NN	O	O
protein	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
genuine	NN	O	O
huGATA-3	NN	O	B-protein
by	NN	O	O
SDS-PAGE	NN	O	O
,	NN	O	O
Western	NN	O	O
blotting	NN	O	O
,	NN	O	O
and	NN	O	O
bandshift	NN	O	O
assays	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1997	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stable	NN	O	O
transfection	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
sense	NN	O	O
or	NN	O	O
antisense	NN	O	B-DNA
RXR-alpha	NN	O	I-DNA
cDNA	NN	O	I-DNA
suggests	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
RXR-alpha	NN	O	B-protein
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
monoblastic	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
and	NN	O	O
vitamin	NN	O	O
D	NN	O	O
.	NN	O	O

Although	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
known	NN	O	O
for	NN	O	O
many	NN	O	O
years	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
modulating	NN	O	O
agent	NN	O	O
that	NN	O	O
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
generating	NN	O	O
both	NN	O	O
granulocytes	NN	O	B-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
underlying	NN	O	O
this	NN	O	O
role	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
defined	NN	O	O
in	NN	O	O
the	NN	O	O
monoblast	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
the	NN	O	O
part	NN	O	O
played	NN	O	O
by	NN	O	O
the	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RXRs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
steroid/thyroid	NN	O	B-protein
hormone	NN	O	I-protein
nuclear	NN	O	I-protein
receptor	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
explored	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monoblastic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U937	NN	O	I-cell_line
has	NN	O	O
been	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
model	NN	O	O
system	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
RXRs	NN	O	B-protein
,	NN	O	O
RXR-alpha	NN	O	B-protein
,	NN	O	O
in	NN	O	O
monoblast	NN	O	O
differentiation	NN	O	O
.	NN	O	O

RXR-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
present	NN	O	O
in	NN	O	O
untreated	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
levels	NN	O	O
increased	NN	O	O
after	NN	O	O
induction	NN	O	O
of	NN	O	O
differentiation	NN	O	O
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
was	NN	O	O
found	NN	O	O
for	NN	O	O
RXR-beta	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Using	NN	O	O
plasmids	NN	O	O
containing	NN	O	O
sense	NN	O	O
or	NN	O	O
antisense	NN	O	B-DNA
RXR-alpha	NN	O	I-DNA
sequences	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	O
promoter	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
stably	NN	O	O
transfected	NN	O	O
cell	NN	O	O
lines	NN	O	O
which	NN	O	O
expressed	NN	O	O
either	NN	O	O
increased	NN	O	O
or	NN	O	O
decreased	NN	O	O
levels	NN	O	O
of	NN	O	O
RXR-alpha	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
sense	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
U	NN	O	B-cell_line
alpha	NN	O	I-cell_line
S	NN	O	I-cell_line
and	NN	O	O
its	NN	O	O
clonal	NN	O	O
derivative	NN	O	O
alpha	NN	O	O
G2S	NN	O	B-cell_line
)	NN	O	O
showed	NN	O	O
increased	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
RA	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
antisense	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
U	NN	O	B-cell_line
alpha	NN	O	I-cell_line
A	NN	O	I-cell_line
and	NN	O	O
its	NN	O	O
clonal	NN	O	O
derivative	NN	O	O
alpha	NN	O	O
B5A	NN	O	B-cell_line
)	NN	O	O
showed	NN	O	O
decreased	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
RA	NN	O	O
,	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
and	NN	O	O
by	NN	O	O
regulation	NN	O	O
of	NN	O	O
an	NN	O	O
RA-responsive	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
U	NN	O	B-cell_line
alpha	NN	O	I-cell_line
A	NN	O	I-cell_line
and	NN	O	O
alpha	NN	O	O
B5A	NN	O	B-cell_line
also	NN	O	O
failed	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
another	NN	O	O
modulating	NN	O	O
agent	NN	O	O
,	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxycholecalciferol	NN	O	O
(	NN	O	O
DHCC	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
U	NN	O	B-cell_line
alpha	NN	O	I-cell_line
S	NN	O	I-cell_line
and	NN	O	O
not	NN	O	O
alpha	NN	O	B-protein
G2S	NN	O	I-protein
showed	NN	O	O
an	NN	O	O
enhanced	NN	O	O
response	NN	O	O
to	NN	O	O
DHCC	NN	O	O
.	NN	O	O

The	NN	O	O
combination	NN	O	O
of	NN	O	O
RA	NN	O	O
and	NN	O	O
DHCC	NN	O	O
together	NN	O	O
inhibited	NN	O	O
growth	NN	O	O
of	NN	O	O
both	NN	O	O
sense	NN	O	O
and	NN	O	O
antisense	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
alpha	NN	O	B-protein
G2S	NN	O	I-protein
exhibited	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
CD11b	NN	O	B-protein
and	NN	O	O
CD54	NN	O	B-protein
,	NN	O	O
while	NN	O	O
alpha	NN	O	B-cell_line
B5A	NN	O	I-cell_line
cells	NN	O	I-cell_line
showed	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
CD102	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
RXR-alpha	NN	O	B-protein
has	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
expression	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
RXR-alpha	NN	O	B-protein
has	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
mediating	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
and	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
during	NN	O	O
myelomonocytic	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
different	NN	O	O
species	NN	O	O
of	NN	O	O
heterodimers	NN	O	B-protein
involving	NN	O	O
RXR-alpha	NN	O	B-protein
may	NN	O	O
control	NN	O	O
the	NN	O	O
acquisition	NN	O	O
of	NN	O	O
different	NN	O	O
features	NN	O	O
of	NN	O	O
mature	NN	O	O
monocyte/macrophage	NN	O	O
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
vascular	NN	O	B-DNA
cell	NN	O	I-DNA
adhesion	NN	O	I-DNA
molecule-1	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
expressing	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
Tax	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Recruitment	NN	O	O
and	NN	O	O
extravasation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
the	NN	O	O
blood-brain	NN	O	O
barrier	NN	O	O
are	NN	O	O
favored	NN	O	O
by	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
-mediated	NN	O	O
interactions	NN	O	O
of	NN	O	O
circulating	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Since	NN	O	O
a	NN	O	O
common	NN	O	O
pathological	NN	O	O
finding	NN	O	O
in	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
-associated	NN	O	O
diseases	NN	O	O
is	NN	O	O
the	NN	O	O
infiltration	NN	O	O
of	NN	O	O
HTLV-1-infected	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
into	NN	O	O
various	NN	O	O
organs	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
looked	NN	O	O
for	NN	O	O
the	NN	O	O
profile	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
expressed	NN	O	O
by	NN	O	O
HTLV-1-transformed	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Flow	NN	O	O
cytometry	NN	O	O
analysis	NN	O	O
indicated	NN	O	O
that	NN	O	O
these	NN	O	O
cells	NN	O	O
were	NN	O	O
expressing	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
[	NN	O	O
CD106	NN	O	B-protein
]	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
110-kDa	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-protein
gene	NN	O	I-protein
superfamily	NN	O	I-protein
,	NN	O	O
first	NN	O	O
identified	NN	O	O
on	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

This	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
was	NN	O	O
also	NN	O	O
expressed	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
one	NN	O	O
patient	NN	O	O
with	NN	O	O
HTLV-1-associated	NN	O	O
myelopathy/tropical	NN	O	O
spastic	NN	O	O
paraparesis	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
one	NN	O	O
normal	NN	O	O
blood	NN	O	O
donor	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
trans-activator	NN	O	I-protein
Tax	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
was	NN	O	O
first	NN	O	O
indicated	NN	O	O
by	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
this	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
on	NN	O	O
Jurkat	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
stably	NN	O	O
expressing	NN	O	O
the	NN	O	O
tax	NN	O	O
gene	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
on	NN	O	O
VCAM-1	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
was	NN	O	O
next	NN	O	O
confirmed	NN	O	O
in	NN	O	O
JPX-9	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
subclone	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
carrying	NN	O	O
the	NN	O	O
tax	NN	O	B-DNA
sequences	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
deletion	NN	O	O
and	NN	O	O
mutation	NN	O	O
analyses	NN	O	O
of	NN	O	O
the	NN	O	O
VCAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
performed	NN	O	O
with	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
constructs	NN	O	I-DNA
revealed	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
was	NN	O	O
trans	NN	O	O
activating	NN	O	O
the	NN	O	O
VCAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
via	NN	O	O
two	NN	O	O
NF-kappaB	NN	O	B-DNA
sites	NN	O	I-DNA
present	NN	O	O
at	NN	O	O
bp	NN	O	O
-72	NN	O	O
and	NN	O	O
-57	NN	O	O
in	NN	O	O
the	NN	O	O
VCAM-1	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
with	NN	O	O
both	NN	O	O
of	NN	O	O
them	NN	O	O
being	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
proteins	NN	O	O
specifically	NN	O	O
bound	NN	O	O
to	NN	O	O
these	NN	O	O
two	NN	O	O
NF-kappaB	NN	O	B-DNA
motifs	NN	O	I-DNA
,	NN	O	O
confirming	NN	O	O
that	NN	O	O
VCAM-1	NN	O	B-protein
was	NN	O	O
induced	NN	O	O
on	NN	O	O
Tax-expressing	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
a	NN	O	O
kappaB	NN	O	B-protein
-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
therefore	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
exclusive	NN	O	O
Tax	NN	O	B-protein
-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
represent	NN	O	O
a	NN	O	O
pivotal	NN	O	O
event	NN	O	O
in	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
HTLV-1-associated	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

HIV-1	NN	O	B-protein
Vpr	NN	O	I-protein
suppresses	NN	O	O
immune	NN	O	O
activation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
through	NN	O	O
regulation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

The	NN	O	O
HIV-1	NN	O	B-protein
accessory	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
Vpr	NN	O	B-protein
can	NN	O	O
influence	NN	O	O
viral	NN	O	O
pathogenesis	NN	O	O
by	NN	O	O
affecting	NN	O	O
viral	NN	O	O
replication	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
host	NN	O	O
cell	NN	O	O
transcription	NN	O	O
and	NN	O	O
proliferation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
Vpr	NN	O	B-protein
on	NN	O	O
host	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
confirm	NN	O	O
that	NN	O	O
it	NN	O	O
influences	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
also	NN	O	O
found	NN	O	O
that	NN	O	O
Vpr	NN	O	B-protein
modulates	NN	O	O
T-cell	NN	O	O
receptor	NN	O	O
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
-triggered	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
a	NN	O	O
manner	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
,	NN	O	O
Vpr	NN	O	B-protein
induces	NN	O	O
apoptosis	NN	O	O
whereas	NN	O	O
in	NN	O	O
its	NN	O	O
presence	NN	O	O
,	NN	O	O
Vpr	NN	O	B-protein
interrupts	NN	O	O
the	NN	O	O
expected	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

This	NN	O	O
regulation	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
linked	NN	O	O
to	NN	O	O
Vpr	NN	O	B-protein
suppression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
via	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Further	NN	O	O
,	NN	O	O
Vpr	NN	O	B-protein
suppresses	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
all	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
Vpr	NN	O	B-protein
could	NN	O	O
be	NN	O	O
reversed	NN	O	O
by	NN	O	O
RU486	NN	O	O
.	NN	O	O

Our	NN	O	O
finding	NN	O	O
that	NN	O	O
Vpr	NN	O	B-protein
can	NN	O	O
regulate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
supports	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
some	NN	O	O
aspects	NN	O	O
of	NN	O	O
viral	NN	O	O
pathogenesis	NN	O	O
are	NN	O	O
the	NN	O	O
consequence	NN	O	O
of	NN	O	O
cell	NN	O	O
dysregulation	NN	O	O
by	NN	O	O
Vpr	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Tissue	NN	O	O
transglutaminase-dependent	NN	O	O
posttranslational	NN	O	O
modification	NN	O	O
of	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
in	NN	O	O
promonocytic	NN	O	B-cell_line
cells	NN	O	I-cell_line
undergoing	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
retinoblastoma	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
(	NN	O	O
pRB	NN	O	B-protein
)	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
controlling	NN	O	O
both	NN	O	O
cell	NN	O	O
release	NN	O	O
from	NN	O	O
the	NN	O	O
G1	NN	O	O
phase	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
early	NN	O	O
phases	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
pRB	NN	O	B-protein
is	NN	O	O
posttranslationally	NN	O	O
modified	NN	O	O
by	NN	O	O
a	NN	O	O
tissue	NN	O	O
transglutaminase	NN	O	O
(	NN	O	O
tTG	NN	O	B-protein
)	NN	O	O
-catalyzed	NN	O	O
reaction	NN	O	O
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
by	NN	O	O
employing	NN	O	O
a	NN	O	O
novel	NN	O	O
haptenized	NN	O	O
lysis	NN	O	O
synthetic	NN	O	O
substrate	NN	O	O
which	NN	O	O
allows	NN	O	O
the	NN	O	O
isolation	NN	O	O
of	NN	O	O
glutaminyl-tTG	NN	O	O
substrates	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
pRB	NN	O	B-protein
as	NN	O	O
a	NN	O	O
potential	NN	O	O
tTG	NN	O	O
substrate	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
undergoing	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
keeping	NN	O	O
with	NN	O	O
this	NN	O	O
finding	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
rapid	NN	O	O
disappearance	NN	O	O
of	NN	O	O
the	NN	O	O
105	NN	O	O
,	NN	O	O
000-	NN	O	O
to	NN	O	O
110	NN	O	O
,	NN	O	O
000-molecular-weight	NN	O	O
pRB	NN	O	B-protein
forms	NN	O	O
concomitantly	NN	O	O
with	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
a	NN	O	O
smear	NN	O	O
of	NN	O	O
immunoreactive	NN	O	O
products	NN	O	O
with	NN	O	O
a	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
greater	NN	O	O
than	NN	O	O
250	NN	O	O
,	NN	O	O
000	NN	O	O
.	NN	O	O

The	NN	O	O
shift	NN	O	O
in	NN	O	O
pRB	NN	O	B-protein
molecular	NN	O	O
weight	NN	O	O
was	NN	O	O
reproduced	NN	O	O
by	NN	O	O
adding	NN	O	O
exogenous	NN	O	O
purified	NN	O	O
tTG	NN	O	O
to	NN	O	O
extracts	NN	O	O
obtained	NN	O	O
from	NN	O	O
viable	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
was	NN	O	O
prevented	NN	O	O
by	NN	O	O
dansylcadaverine	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
enzyme	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
pRB	NN	O	B-protein
posttranslational	NN	O	O
modification	NN	O	O
during	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
investigated	NN	O	O
by	NN	O	O
determining	NN	O	O
the	NN	O	O
E2F-1	NN	O	B-protein
levels	NN	O	O
and	NN	O	O
by	NN	O	O
isolating	NN	O	O
and	NN	O	O
characterizing	NN	O	O
pRB-null	NN	O	B-cell_line
clones	NN	O	I-cell_line
from	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Notably	NN	O	O
,	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
pRB	NN	O	B-protein
in	NN	O	O
these	NN	O	O
U937-derived	NN	O	B-cell_line
clones	NN	O	I-cell_line
renders	NN	O	O
these	NN	O	O
p53-null	NN	O	B-cell_line
cells	NN	O	I-cell_line
highly	NN	O	O
resistant	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
serum	NN	O	O
withdrawal	NN	O	O
,	NN	O	O
calphostin	NN	O	O
C	NN	O	O
,	NN	O	O
and	NN	O	O
ceramide	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
tTG	NN	O	B-protein
,	NN	O	O
acting	NN	O	O
on	NN	O	O
the	NN	O	O
pRB	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
might	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
progression	NN	O	O
through	NN	O	O
the	NN	O	O
death	NN	O	O
program	NN	O	O
.	NN	O	O

-DOCSTART-	O

STAT3	NN	O	B-protein
is	NN	O	O
a	NN	O	O
serine	NN	O	B-protein
kinase	NN	O	I-protein
target	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Interleukin	NN	O	B-protein
2	NN	O	I-protein
and	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
signals	NN	O	O
converge	NN	O	O
upon	NN	O	O
serine	NN	O	O
727	NN	O	O
.	NN	O	O

Interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STATs	NN	O	B-protein
3	NN	O	I-protein
and	NN	O	I-protein
5	NN	O	I-protein
(	NN	O	O
signal	NN	O	O
transducer	NN	O	O
and	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
regulation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
a	NN	O	O
complex	NN	O	O
response	NN	O	O
involving	NN	O	O
activation	NN	O	O
of	NN	O	O
two	NN	O	O
forms	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
:	NN	O	O
90-kDa	NN	O	O
STAT3alpha	NN	O	B-protein
and	NN	O	O
an	NN	O	O
83-kDa	NN	O	B-protein
carboxyl-terminal	NN	O	I-protein
truncated	NN	O	I-protein
STAT3beta	NN	O	I-protein
.	NN	O	O

The	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
on	NN	O	O
serine	NN	O	O
residues	NN	O	O
is	NN	O	O
also	NN	O	O
required	NN	O	O
for	NN	O	O
competent	NN	O	O
STAT	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

A	NN	O	O
critical	NN	O	O
serine	NN	O	B-protein
phosphorylation	NN	O	I-protein
site	NN	O	I-protein
in	NN	O	O
STAT3alpha	NN	O	B-protein
is	NN	O	O
at	NN	O	O
position	NN	O	O
727	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
have	NN	O	O
produced	NN	O	O
an	NN	O	O
antisera	NN	O	O
specific	NN	O	O
for	NN	O	O
STAT3alpha	NN	O	B-protein
proteins	NN	O	I-protein
phosphorylated	NN	O	O
on	NN	O	O
serine	NN	O	O
727	NN	O	O
and	NN	O	O
used	NN	O	O
this	NN	O	O
to	NN	O	O
monitor	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
this	NN	O	O
residue	NN	O	O
during	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT3alpha	NN	O	B-protein
on	NN	O	O
serine	NN	O	O
727	NN	O	O
is	NN	O	O
not	NN	O	O
constitutive	NN	O	O
in	NN	O	O
quiescent	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
IL-2	NN	O	I-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
triggering	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
or	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
also	NN	O	O
induces	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
serine	NN	O	O
727	NN	O	O
but	NN	O	O
without	NN	O	O
simultaneously	NN	O	O
inducing	NN	O	O
STAT3	NN	O	B-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
or	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
the	NN	O	O
present	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
STAT3	NN	O	B-protein
serine	NN	O	O
phosphorylation	NN	O	O
can	NN	O	O
be	NN	O	O
regulated	NN	O	O
independently	NN	O	O
of	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
this	NN	O	O
molecule	NN	O	O
.	NN	O	O

IL-2	NN	O	B-protein
and	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
induction	NN	O	O
of	NN	O	O
STAT3alpha	NN	O	B-protein
serine	NN	O	O
727	NN	O	O
phosphorylation	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
MEK	NN	O	B-protein
/ERK	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
identified	NN	O	O
H-7-sensitive	NN	O	O
kinase	NN	O	O
pathways	NN	O	O
that	NN	O	O
regulate	NN	O	O
STAT3	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
H-7-sensitive	NN	O	O
pathways	NN	O	O
regulate	NN	O	O
STAT3	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
H-7-sensitive	NN	O	B-protein
kinases	NN	O	I-protein
do	NN	O	O
not	NN	O	O
regulate	NN	O	O
STAT3	NN	O	B-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
or	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
serine	NN	O	O
727	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
thus	NN	O	O
show	NN	O	O
that	NN	O	O
STAT3	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
targets	NN	O	O
for	NN	O	O
multiple	NN	O	O
kinase	NN	O	O
pathways	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
can	NN	O	O
integrate	NN	O	O
signals	NN	O	O
from	NN	O	O
both	NN	O	O
cytokine	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
antigen	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
regulating	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
monocytic	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
kappa	NN	O	B-DNA
B-site	NN	O	I-DNA
was	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	O
of	NN	O	O
the	NN	O	O
intercellular	NN	O	B-DNA
adhesion	NN	O	I-DNA
molecule-1	NN	O	I-DNA
(	NN	O	I-DNA
ICAM-1	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
on	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
which	NN	O	O
bind	NN	O	O
and	NN	O	O
transactivate	NN	O	O
this	NN	O	O
enhancer	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
,	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
appeared	NN	O	O
to	NN	O	O
bind	NN	O	O
RelA	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
homodimers	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
heterodimers	NN	O	O
with	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
,	NN	O	O
but	NN	O	O
weakly	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
homodimers	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
both	NN	O	O
RelA	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
,	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
transactivate	NN	O	O
an	NN	O	O
ICAM-1	NN	O	B-DNA
kappa	NN	O	I-DNA
B-reporter	NN	O	I-DNA
construct	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
monocytic	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
TNF-alpha	NN	O	B-protein
induced	NN	O	O
two	NN	O	O
nuclear	NN	O	B-protein
complexes	NN	O	I-protein
which	NN	O	O
in	NN	O	O
vitro	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
antibodies	NN	O	O
in	NN	O	O
an	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
supershift	NN	O	O
assay	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
complexes	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
contain	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
and	NN	O	O
RelA	NN	O	B-protein
,	NN	O	O
and	NN	O	O
to	NN	O	O
bind	NN	O	O
with	NN	O	O
higher	NN	O	O
affinity	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
second	NN	O	O
complex	NN	O	O
contained	NN	O	O
RelA	NN	O	B-protein
,	NN	O	O
and	NN	O	O
exhibited	NN	O	O
higher	NN	O	O
affinity	NN	O	O
towards	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
than	NN	O	O
to	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
repress	NN	O	O
activity	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
RelA	NN	O	B-protein
homodimer	NN	O	I-protein
and	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B1/RelA	NN	O	I-protein
heterodimer	NN	O	I-protein
.	NN	O	O

We	NN	O	O
argue	NN	O	O
that	NN	O	O
in	NN	O	O
vivo	NN	O	O
RelA	NN	O	B-protein
homodimers	NN	O	I-protein
are	NN	O	O
likely	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
dominant	NN	O	O
role	NN	O	O
in	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
ICAM-1	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Competent	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
by	NN	O	O
RNA	NN	O	B-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
in	NN	O	O
cell-free	NN	O	B-cell_type
extracts	NN	O	I-cell_type
from	NN	O	O
xeroderma	NN	O	O
pigmentosum	NN	O	O
groups	NN	O	O
B	NN	O	O
and	NN	O	O
D	NN	O	O
in	NN	O	O
an	NN	O	O
optimized	NN	O	O
RNA	NN	O	O
transcription	NN	O	O
assay	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
disease	NN	O	O
,	NN	O	O
xeroderma	NN	O	O
pigmentosum	NN	O	O
(	NN	O	O
XP	NN	O	O
)	NN	O	O
,	NN	O	O
can	NN	O	O
result	NN	O	O
from	NN	O	O
mutations	NN	O	O
in	NN	O	O
any	NN	O	O
one	NN	O	O
of	NN	O	O
seven	NN	O	O
genes	NN	O	O
,	NN	O	O
designated	NN	O	O
XPA	NN	O	B-DNA
through	NN	O	O
XPG	NN	O	B-DNA
.	NN	O	O

Of	NN	O	O
these	NN	O	O
,	NN	O	O
the	NN	O	O
XPB	NN	O	B-DNA
and	NN	O	I-DNA
XPD	NN	O	I-DNA
genes	NN	O	I-DNA
encode	NN	O	O
proteins	NN	O	O
that	NN	O	O
are	NN	O	O
subunits	NN	O	O
of	NN	O	O
a	NN	O	O
general	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
TFIIH	NN	O	B-protein
,	NN	O	O
involved	NN	O	O
in	NN	O	O
both	NN	O	O
nucleotide	NN	O	O
excision	NN	O	O
repair	NN	O	O
(	NN	O	O
NER	NN	O	O
)	NN	O	O
and	NN	O	O
initiation	NN	O	O
of	NN	O	O
mRNA	NN	O	O
transcription	NN	O	O
by	NN	O	O
RNA	NN	O	B-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
.	NN	O	O

In	NN	O	O
humans	NN	O	O
,	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
XPB	NN	O	B-DNA
or	NN	O	I-DNA
XPD	NN	O	I-DNA
gene	NN	O	I-DNA
impairs	NN	O	O
NER	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
hyper-sensitivity	NN	O	O
to	NN	O	O
sunlight	NN	O	O
and	NN	O	O
greatly	NN	O	O
increased	NN	O	O
skin	NN	O	O
tumor	NN	O	O
formation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
no	NN	O	O
transcription	NN	O	O
deficiency	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
either	NN	O	O
XP-B	NN	O	O
or	NN	O	O
XP-D	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
employed	NN	O	O
an	NN	O	O
optimized	NN	O	O
cell-free	NN	O	O
RNA	NN	O	O
transcription	NN	O	O
assay	NN	O	O
to	NN	O	O
analyze	NN	O	O
transcription	NN	O	O
activity	NN	O	O
of	NN	O	O
XP-B	NN	O	O
and	NN	O	O
XP-D	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
growth	NN	O	O
rate	NN	O	O
was	NN	O	O
normal	NN	O	O
,	NN	O	O
the	NN	O	O
XP-B	NN	O	B-cell_line
and	NN	O	I-cell_line
XP-D	NN	O	I-cell_line
cells	NN	O	I-cell_line
contained	NN	O	O
reduced	NN	O	O
amounts	NN	O	O
of	NN	O	O
TFIIH	NN	O	B-protein
.	NN	O	O

Extracts	NN	O	O
prepared	NN	O	O
from	NN	O	O
XP-B	NN	O	B-cell_line
and	NN	O	I-cell_line
XP-D	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
exhibited	NN	O	O
similar	NN	O	O
transcription	NN	O	O
activity	NN	O	O
from	NN	O	O
the	NN	O	O
adenovirus	NN	O	B-DNA
major	NN	O	I-DNA
late	NN	O	I-DNA
promoter	NN	O	I-DNA
when	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
in	NN	O	O
extracts	NN	O	O
from	NN	O	O
normal	NN	O	O
cells	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
XP-B	NN	O	B-cell_line
and	NN	O	I-cell_line
XP-D	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
do	NN	O	O
not	NN	O	O
have	NN	O	O
impaired	NN	O	O
RNA	NN	O	O
transcription	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
consider	NN	O	O
the	NN	O	O
possible	NN	O	O
consequences	NN	O	O
of	NN	O	O
the	NN	O	O
reduced	NN	O	O
cellular	NN	O	O
content	NN	O	O
of	NN	O	O
TFIIH	NN	O	B-protein
for	NN	O	O
the	NN	O	O
clinical	NN	O	O
symptoms	NN	O	O
in	NN	O	O
XP-B	NN	O	O
or	NN	O	O
XP-D	NN	O	O
patients	NN	O	O
,	NN	O	O
and	NN	O	O
discuss	NN	O	O
a	NN	O	O
'conditional	NN	O	O
phenotype	NN	O	O
'	NN	O	O
that	NN	O	O
may	NN	O	O
involve	NN	O	O
an	NN	O	O
impairment	NN	O	O
of	NN	O	O
cellular	NN	O	O
function	NN	O	O
only	NN	O	O
under	NN	O	O
certain	NN	O	O
growth	NN	O	O
conditions	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
myeloid-specific	NN	O	B-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
MZF-2	NN	O	B-protein
)	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Myeloid	NN	O	O
cell	NN	O	O
development	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
tissue-specific	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Human	NN	O	B-protein
myeloid	NN	O	I-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
MZF-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
putative	NN	O	O
transcription	NN	O	O
factor	NN	O	O
containing	NN	O	O
13	NN	O	B-protein
zinc	NN	O	I-protein
fingers	NN	O	I-protein
,	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
that	NN	O	O
it	NN	O	O
regulates	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
neutrophilic	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Here	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
isolated	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
and	NN	O	I-DNA
human	NN	O	I-DNA
cDNAs	NN	O	I-DNA
which	NN	O	O
encode	NN	O	O
a	NN	O	O
novel	NN	O	O
form	NN	O	O
of	NN	O	O
MZF	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
MZF-2	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Murine	NN	O	B-protein
and	NN	O	I-protein
human	NN	O	I-protein
MZF-2	NN	O	I-protein
proteins	NN	O	I-protein
consisted	NN	O	O
of	NN	O	O
814	NN	O	O
and	NN	O	O
775	NN	O	O
amino	NN	O	O
acids	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
and	NN	O	O
have	NN	O	O
identity	NN	O	O
of	NN	O	O
75.3	NN	O	O
%	NN	O	O
between	NN	O	O
them	NN	O	O
.	NN	O	O

The	NN	O	O
C-terminal	NN	O	B-protein
half	NN	O	I-protein
of	NN	O	O
human	NN	O	B-protein
MZF-2	NN	O	I-protein
,	NN	O	O
carrying	NN	O	O
the	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
domains	NN	O	I-protein
,	NN	O	O
was	NN	O	O
completely	NN	O	O
identical	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
human	NN	O	B-protein
MZF-1	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
half	NN	O	I-protein
of	NN	O	O
MZF-2	NN	O	B-protein
was	NN	O	O
different	NN	O	O
from	NN	O	O
the	NN	O	O
corresponding	NN	O	O
region	NN	O	O
of	NN	O	O
human	NN	O	O
MZF-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
was	NN	O	O
coded	NN	O	O
by	NN	O	O
distinct	NN	O	O
exons	NN	O	B-DNA
.	NN	O	O

MZF-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
myeloid	NN	O	O
cells	NN	O	O
,	NN	O	O
particularly	NN	O	O
in	NN	O	O
the	NN	O	O
cells	NN	O	O
committed	NN	O	O
to	NN	O	O
the	NN	O	O
neutrophilic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
down-regulated	NN	O	O
by	NN	O	O
G-CSF	NN	O	B-protein
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
MZF-1	NN	O	B-RNA
and	NN	O	I-RNA
MZF-2	NN	O	I-RNA
mRNAs	NN	O	I-RNA
seem	NN	O	O
to	NN	O	O
be	NN	O	O
produced	NN	O	O
by	NN	O	O
the	NN	O	O
alternative	NN	O	O
use	NN	O	O
of	NN	O	O
two	NN	O	O
different	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
distinct	NN	O	O
N-terminal	NN	O	B-protein
half	NN	O	I-protein
of	NN	O	O
MZF-2	NN	O	B-protein
carries	NN	O	O
two	NN	O	O
characteristic	NN	O	O
domains	NN	O	O
,	NN	O	O
a	NN	O	O
leucine-rich	NN	O	B-protein
domain	NN	O	I-protein
called	NN	O	O
LeR	NN	O	B-protein
and	NN	O	O
an	NN	O	O
acidic	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
which	NN	O	O
suggests	NN	O	O
a	NN	O	O
unique	NN	O	O
function	NN	O	O
of	NN	O	O
MZF-2	NN	O	B-protein
in	NN	O	O
neutrophil	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Impaired	NN	O	O
cortisol	NN	O	O
binding	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
hypertensive	NN	O	O
patients	NN	O	O
.	NN	O	O

We	NN	O	O
compared	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
HML	NN	O	B-cell_type
)	NN	O	O
from	NN	O	O
hypertensive	NN	O	O
patients	NN	O	O
and	NN	O	O
matched	NN	O	O
normotensive	NN	O	O
volunteers	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
considered	NN	O	O
associations	NN	O	O
of	NN	O	O
these	NN	O	O
variables	NN	O	O
with	NN	O	O
plasma	NN	O	O
renin	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
aldosterone	NN	O	O
,	NN	O	O
cortisol	NN	O	O
,	NN	O	O
corticotropin	NN	O	O
,	NN	O	O
and	NN	O	O
electrolyte	NN	O	O
concentrations	NN	O	O
.	NN	O	O

We	NN	O	O
calculated	NN	O	O
binding	NN	O	O
affinity	NN	O	O
(	NN	O	O
Kd	NN	O	O
;	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
and	NN	O	O
capacity	NN	O	O
(	NN	O	O
Bmax	NN	O	O
;	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
cortisol	NN	O	O
from	NN	O	O
homologous	NN	O	O
and	NN	O	O
heterologous	NN	O	O
competition	NN	O	O
curves	NN	O	O
for	NN	O	O
specific	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
sites	NN	O	O
on	NN	O	O
HML	NN	O	B-cell_type
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
blood	NN	O	O
of	NN	O	O
normotensive	NN	O	O
volunteers	NN	O	O
and	NN	O	O
subjects	NN	O	O
with	NN	O	O
essential	NN	O	O
hypertension	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
HML	NN	O	B-cell_type
was	NN	O	O
evaluated	NN	O	O
as	NN	O	O
IC50	NN	O	O
values	NN	O	O
(	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
cortisol	NN	O	O
for	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
lysozyme	NN	O	B-protein
release	NN	O	O
.	NN	O	O

We	NN	O	O
measured	NN	O	O
plasma	NN	O	O
hormones	NN	O	O
by	NN	O	O
radioimmunoassay	NN	O	O
.	NN	O	O

Kd	NN	O	O
values	NN	O	O
(	NN	O	O
mean+/-SE	NN	O	O
)	NN	O	O
for	NN	O	O
cortisol	NN	O	O
in	NN	O	O
HML	NN	O	B-cell_type
of	NN	O	O
hypertensive	NN	O	O
patients	NN	O	O
were	NN	O	O
higher	NN	O	O
than	NN	O	O
in	NN	O	O
control	NN	O	O
subjects	NN	O	O
(	NN	O	O
24.6+/-2.4	NN	O	O
versus	NN	O	O
17.5+/-1.7	NN	O	O
nmol/L	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
.04	NN	O	O
)	NN	O	O
.	NN	O	O

Binding	NN	O	O
capacity	NN	O	O
(	NN	O	O
4978+/-391	NN	O	O
versus	NN	O	O
4131+/-321	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
,	NN	O	O
Kd	NN	O	O
values	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
6.7+/-0.5	NN	O	O
versus	NN	O	O
5.7+/-0.3	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
IC50	NN	O	O
values	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
3.4+/-0.3	NN	O	O
versus	NN	O	O
3.1+/-0.2	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
and	NN	O	O
cortisol	NN	O	O
(	NN	O	O
12.2+/-1.6	NN	O	O
versus	NN	O	O
9.5+/-0.3	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
were	NN	O	O
not	NN	O	O
significantly	NN	O	O
different	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
renin	NN	O	B-protein
values	NN	O	O
less	NN	O	O
than	NN	O	O
0.13	NN	O	O
ng	NN	O	O
angiotensin	NN	O	O
I/L	NN	O	O
per	NN	O	O
second	NN	O	O
were	NN	O	O
markedly	NN	O	O
less	NN	O	O
sensitive	NN	O	O
to	NN	O	O
cortisol	NN	O	O
than	NN	O	O
those	NN	O	O
with	NN	O	O
higher	NN	O	O
values	NN	O	O
.	NN	O	O

Both	NN	O	O
Kd	NN	O	O
(	NN	O	O
30.3+/-2.5	NN	O	O
versus	NN	O	O
19.2+/-2.4	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
and	NN	O	O
IC50	NN	O	O
values	NN	O	O
(	NN	O	O
15.5+/-1.8	NN	O	O
versus	NN	O	O
8.9+/-1.2	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
for	NN	O	O
cortisol	NN	O	O
were	NN	O	O
significantly	NN	O	O
higher	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
lower	NN	O	O
renin	NN	O	B-protein
values	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
.03	NN	O	O
)	NN	O	O
.	NN	O	O

Other	NN	O	O
variables	NN	O	O
,	NN	O	O
including	NN	O	O
plasma	NN	O	O
hormone	NN	O	O
and	NN	O	O
electrolyte	NN	O	O
values	NN	O	O
and	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
were	NN	O	O
not	NN	O	O
different	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
cortisol	NN	O	O
binding	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
slightly	NN	O	O
impaired	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
essential	NN	O	O
hypertension	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
,	NN	O	O
this	NN	O	O
could	NN	O	O
lead	NN	O	O
to	NN	O	O
inappropriate	NN	O	O
binding	NN	O	O
of	NN	O	O
cortisol	NN	O	O
to	NN	O	O
mineralocorticoid	NN	O	O
receptors	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
decreased	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
cortisol	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
renin	NN	O	B-protein
suppression	NN	O	O
.	NN	O	O

This	NN	O	O
hypothesis	NN	O	O
is	NN	O	O
supported	NN	O	O
by	NN	O	O
evidence	NN	O	O
of	NN	O	O
hypertension	NN	O	O
and	NN	O	O
low	NN	O	O
renin	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
which	NN	O	O
others	NN	O	O
have	NN	O	O
described	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
primary	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
due	NN	O	O
to	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

PU.1/Pip	NN	O	B-protein
and	NN	O	O
basic	NN	O	B-protein
helix	NN	O	I-protein
loop	NN	O	I-protein
helix	NN	O	I-protein
zipper	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
interact	NN	O	O
with	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
CD20	NN	O	B-DNA
promoter	NN	O	I-DNA
to	NN	O	O
help	NN	O	O
confer	NN	O	O
lineage-	NN	O	O
and	NN	O	O
stage-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
CD20	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

CD20	NN	O	B-protein
is	NN	O	O
a	NN	O	O
B-lineage-specific	NN	O	B-DNA
gene	NN	O	I-DNA
expressed	NN	O	O
at	NN	O	O
the	NN	O	O
pre-B-cell	NN	O	O
stage	NN	O	O
of	NN	O	O
B-cell	NN	O	O
development	NN	O	O
that	NN	O	O
disappears	NN	O	O
on	NN	O	O
differentiation	NN	O	O
to	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

As	NN	O	O
such	NN	O	O
,	NN	O	O
it	NN	O	O
serves	NN	O	O
as	NN	O	O
an	NN	O	O
excellent	NN	O	O
paradigm	NN	O	O
for	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
lineage	NN	O	O
and	NN	O	O
developmental	NN	O	O
stage-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Using	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprinting	NN	O	O
we	NN	O	O
identified	NN	O	O
two	NN	O	O
sites	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	O
at	NN	O	O
-45	NN	O	B-DNA
and	NN	O	I-DNA
-160	NN	O	I-DNA
that	NN	O	O
were	NN	O	O
occupied	NN	O	O
only	NN	O	O
in	NN	O	O
CD20	NN	O	B-protein
+	NN	O	O
B	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
-45	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
an	NN	O	O
E	NN	O	B-DNA
box	NN	O	I-DNA
that	NN	O	O
binds	NN	O	O
basic	NN	O	B-protein
helix-loop-helix-zipper	NN	O	I-protein
proteins	NN	O	I-protein
whereas	NN	O	O
the	NN	O	O
-160	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
composite	NN	O	B-DNA
PU.1	NN	O	I-DNA
and	NN	O	I-DNA
Pip	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Transfection	NN	O	O
studies	NN	O	O
with	NN	O	O
reporter	NN	O	B-DNA
constructs	NN	O	I-DNA
and	NN	O	O
various	NN	O	O
expression	NN	O	O
vectors	NN	O	O
verified	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
these	NN	O	O
sites	NN	O	O
.	NN	O	O

The	NN	O	O
composite	NN	O	B-DNA
PU.1	NN	O	I-DNA
and	NN	O	I-DNA
Pip	NN	O	I-DNA
site	NN	O	I-DNA
likely	NN	O	O
accounts	NN	O	O
for	NN	O	O
both	NN	O	O
lineage	NN	O	O
and	NN	O	O
stage-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
CD20	NN	O	B-protein
whereas	NN	O	O
the	NN	O	O
CD20	NN	O	B-protein
E	NN	O	I-protein
box	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
enhance	NN	O	O
overall	NN	O	O
promoter	NN	O	O
activity	NN	O	O
and	NN	O	O
may	NN	O	O
link	NN	O	O
the	NN	O	O
promoter	NN	O	O
to	NN	O	O
a	NN	O	O
distant	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Glucocorticoids	NN	O	O
and	NN	O	O
the	NN	O	O
immune	NN	O	O
function	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
infection	NN	O	O
:	NN	O	O
a	NN	O	O
study	NN	O	O
in	NN	O	O
hypercortisolemic	NN	O	O
and	NN	O	O
cortisol-resistant	NN	O	O
patients	NN	O	O
.	NN	O	O

Immunological	NN	O	O
studies	NN	O	O
in	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
-positive	NN	O	O
patients	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
disease	NN	O	O
progression	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
defective	NN	O	O
production	NN	O	O
of	NN	O	O
type	NN	O	O
1	NN	O	B-protein
cytokines	NN	O	I-protein
(	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IL-12	NN	O	B-protein
]	NN	O	O
,	NN	O	O
an	NN	O	O
increased	NN	O	O
production	NN	O	O
of	NN	O	O
type	NN	O	B-protein
2	NN	O	I-protein
cytokines	NN	O	I-protein
(	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
an	NN	O	O
increased	NN	O	O
production	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
.	NN	O	O

HIV	NN	O	O
infection	NN	O	O
is	NN	O	O
also	NN	O	O
associated	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
hypothalamo-pituitary-adrenal	NN	O	O
axis	NN	O	O
function	NN	O	O
and	NN	O	O
increased	NN	O	O
plasma	NN	O	O
and	NN	O	O
urinary	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
.	NN	O	O

As	NN	O	O
cortisol	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
physiological	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
a	NN	O	O
study	NN	O	O
was	NN	O	O
conducted	NN	O	O
to	NN	O	O
examine	NN	O	O
cytokine	NN	O	B-protein
patterns	NN	O	O
in	NN	O	O
two	NN	O	O
groups	NN	O	O
of	NN	O	O
hypercortisolemic	NN	O	O
patients	NN	O	O
,	NN	O	O
one	NN	O	O
with	NN	O	O
normal	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
the	NN	O	O
other	NN	O	O
with	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
.	NN	O	O

Ten	NN	O	O
HIV-infected	NN	O	O
patients	NN	O	O
with	NN	O	O
normal	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
(	NN	O	O
AIDS-C	NN	O	O
)	NN	O	O
,	NN	O	O
10	NN	O	O
HIV-infected	NN	O	O
patients	NN	O	O
with	NN	O	O
low	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
(	NN	O	O
AIDS-GR	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
20	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

Receptor	NN	O	O
characteristics	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
evaluated	NN	O	O
by	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
.	NN	O	O

Serum	NN	O	O
cortisol	NN	O	O
and	NN	O	O
urinary	NN	O	O
free	NN	O	O
cortisol	NN	O	O
were	NN	O	O
measured	NN	O	O
by	NN	O	O
RIA	NN	O	O
.	NN	O	O

Serum	NN	O	O
ACTH	NN	O	O
and	NN	O	O
IgE	NN	O	B-protein
were	NN	O	O
measured	NN	O	O
by	NN	O	O
immunoradiometric	NN	O	O
assay	NN	O	O
,	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-10	NN	O	B-protein
cytokines	NN	O	B-protein
and	NN	O	O
interferon-gamma	NN	O	B-protein
were	NN	O	O
measured	NN	O	O
by	NN	O	O
enzyme-linked	NN	O	O
immunosorbent	NN	O	O
assay	NN	O	O
.	NN	O	O

AIDS-C	NN	O	O
patients	NN	O	O
showed	NN	O	O
low	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
high	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IgE	NN	O	B-protein
concentratios	NN	O	O
;	NN	O	O
conversely	NN	O	O
,	NN	O	O
AIDS-GR	NN	O	O
patients	NN	O	O
showed	NN	O	O
high	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
low	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IgE	NN	O	B-protein
concentrations	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
in	NN	O	O
HIV	NN	O	O
infection	NN	O	O
,	NN	O	O
elevated	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
suppress	NN	O	O
cell-mediated	NN	O	O
immunity	NN	O	O
and	NN	O	O
stimulate	NN	O	O
humoral	NN	O	O
immunity	NN	O	O
,	NN	O	O
whereas	NN	O	O
this	NN	O	O
response	NN	O	O
is	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
cortisol-resistant	NN	O	O
patients	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
cortisol	NN	O	O
and	NN	O	O
its	NN	O	O
receptors	NN	O	O
are	NN	O	O
critically	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
immune	NN	O	O
function	NN	O	O
in	NN	O	O
HIV	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
Bcl-xL	NN	O	B-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
ultimately	NN	O	O
induces	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
along	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
treatment	NN	O	O
with	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
or	NN	O	O
DMSO	NN	O	O
induces	NN	O	O
granulocytic/neutrophillic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
disparate	NN	O	O
fates	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
treated	NN	O	O
with	NN	O	O
the	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
12	NN	O	O
,	NN	O	O
13-phorbol	NN	O	O
dibutyric	NN	O	O
acid	NN	O	O
(	NN	O	O
PDBu	NN	O	O
)	NN	O	O
or	NN	O	O
DMSO	NN	O	O
.	NN	O	O

After	NN	O	O
DMSO	NN	O	O
treatment	NN	O	O
,	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
eventually	NN	O	O
died	NN	O	O
via	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
viability	NN	O	O
of	NN	O	O
PDBu-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
not	NN	O	O
affected	NN	O	O
during	NN	O	O
the	NN	O	O
same	NN	O	O
interval	NN	O	O
.	NN	O	O

The	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
apoptosis	NN	O	O
effector	NN	O	O
proteins	NN	O	O
Bak	NN	O	B-protein
and	NN	O	O
Bad	NN	O	B-protein
were	NN	O	O
enhanced	NN	O	O
,	NN	O	O
whereas	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
slight	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
apoptosis	NN	O	B-protein
suppressor	NN	O	I-protein
protein	NN	O	I-protein
Bcl-2	NN	O	B-protein
after	NN	O	O
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
PDBu	NN	O	O
and	NN	O	O
DMSO	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
DMSO	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
elevation	NN	O	O
of	NN	O	O
the	NN	O	O
apoptosis	NN	O	B-protein
effector	NN	O	I-protein
Bax	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
treatment	NN	O	O
with	NN	O	O
PDBu	NN	O	O
did	NN	O	O
not	NN	O	O
significantly	NN	O	O
alter	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
treatment	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
PDBu	NN	O	O
induced	NN	O	O
the	NN	O	O
rapid	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
apoptosis	NN	O	B-protein
suppressor	NN	O	I-protein
protein	NN	O	I-protein
Bcl-xL	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
remained	NN	O	O
unaltered	NN	O	O
in	NN	O	O
DMSO-treated	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
generality	NN	O	O
of	NN	O	O
this	NN	O	O
finding	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
Bcl-xL	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
U-937	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
exposed	NN	O	O
to	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
,	NN	O	O
and	NN	O	O
mouse	NN	O	B-cell_line
thioglycollate-activated	NN	O	I-cell_line
and	NN	O	I-cell_line
resident	NN	O	I-cell_line
peritoneal	NN	O	I-cell_line
macrophages	NN	O	I-cell_line
.	NN	O	O

PDBu-treated	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
remained	NN	O	O
viable	NN	O	O
for	NN	O	O
7	NN	O	O
days	NN	O	O
and	NN	O	O
thereafter	NN	O	O
began	NN	O	O
to	NN	O	O
die	NN	O	O
via	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
concomitant	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
Bcl-xL	NN	O	B-protein
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
Bcl-xL	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
differentiation	NN	O	O
and	NN	O	O
survival	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
along	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Monochloramine	NN	O	O
inhibits	NN	O	O
phorbol	NN	O	O
ester-inducible	NN	O	O
neutrophil	NN	O	O
respiratory	NN	O	O
burst	NN	O	O
activation	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	O
expression	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
inducible	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

Monochloramine	NN	O	O
derivatives	NN	O	O
are	NN	O	O
long	NN	O	O
lived	NN	O	O
physiological	NN	O	O
oxidants	NN	O	O
produced	NN	O	O
by	NN	O	O
neutrophils	NN	O	B-cell_type
during	NN	O	O
the	NN	O	O
respiratory	NN	O	O
burst	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
chemically	NN	O	O
prepared	NN	O	O
monochloramine	NN	O	O
(	NN	O	O
NH2Cl	NN	O	O
)	NN	O	O
on	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
and	NN	O	O
PKC	NN	O	B-protein
-mediated	NN	O	O
cellular	NN	O	O
responses	NN	O	O
were	NN	O	O
studied	NN	O	O
in	NN	O	O
elicited	NN	O	B-cell_type
rat	NN	O	I-cell_type
peritoneal	NN	O	I-cell_type
neutrophils	NN	O	I-cell_type
and	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Neutrophils	NN	O	O
pretreated	NN	O	O
with	NN	O	O
NH2Cl	NN	O	O
(	NN	O	O
30-50	NN	O	O
microM	NN	O	O
)	NN	O	O
showed	NN	O	O
a	NN	O	O
marked	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
respiratory	NN	O	O
burst	NN	O	O
activity	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
PKC	NN	O	B-protein
activator	NN	O	O
.	NN	O	O

These	NN	O	O
cells	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
were	NN	O	O
viable	NN	O	O
and	NN	O	O
showed	NN	O	O
a	NN	O	O
complete	NN	O	O
respiratory	NN	O	O
burst	NN	O	O
upon	NN	O	O
arachidonic	NN	O	O
acid	NN	O	O
stimulation	NN	O	O
,	NN	O	O
which	NN	O	O
induces	NN	O	O
the	NN	O	O
respiratory	NN	O	O
burst	NN	O	O
by	NN	O	O
a	NN	O	O
PKC	NN	O	B-protein
-independent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

The	NN	O	O
NH2Cl-treated	NN	O	B-cell_line
neutrophils	NN	O	I-cell_line
showed	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
both	NN	O	O
PKC	NN	O	B-protein
activity	NN	O	O
and	NN	O	O
PMA-induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
a	NN	O	O
47-kDa	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
corresponds	NN	O	O
to	NN	O	O
the	NN	O	O
cytosolic	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	O
NADPH	NN	O	B-protein
oxidase	NN	O	I-protein
,	NN	O	O
p47	NN	O	B-protein
(	NN	O	I-protein
phox	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
pretreated	NN	O	O
with	NN	O	O
NH2Cl	NN	O	O
(	NN	O	O
20-70	NN	O	O
microM	NN	O	O
)	NN	O	O
showed	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
following	NN	O	O
PMA	NN	O	O
stimulation	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
also	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
both	NN	O	O
PKC	NN	O	B-protein
activity	NN	O	O
and	NN	O	O
nuclear	NN	O	O
transcription	NN	O	O
factor-	NN	O	O
kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
also	NN	O	O
without	NN	O	O
loss	NN	O	O
of	NN	O	O
cell	NN	O	O
viability	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
NH2Cl	NN	O	O
inhibits	NN	O	O
PKC	NN	O	B-protein
-mediated	NN	O	O
cellular	NN	O	O
responses	NN	O	O
through	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	O
PKC	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Evidence	NN	O	O
that	NN	O	O
calcineurin	NN	O	O
is	NN	O	O
rate-limiting	NN	O	O
for	NN	O	O
primary	NN	O	O
human	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

Cyclosporine	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
is	NN	O	O
both	NN	O	O
a	NN	O	O
clinical	NN	O	O
immunosuppressive	NN	O	O
drug	NN	O	O
and	NN	O	O
a	NN	O	O
probe	NN	O	O
to	NN	O	O
dissect	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
,	NN	O	O
CsA	NN	O	O
inhibits	NN	O	O
lymphocyte	NN	O	B-DNA
gene	NN	O	I-DNA
activation	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
phosphatase	NN	O	B-protein
activity	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
(	NN	O	O
CN	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
clinical	NN	O	O
use	NN	O	O
,	NN	O	O
CsA	NN	O	O
treatment	NN	O	O
inhibits	NN	O	O
50-75	NN	O	O
%	NN	O	O
of	NN	O	O
CN	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
circulating	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
modeled	NN	O	O
this	NN	O	O
degree	NN	O	O
of	NN	O	O
CN	NN	O	B-protein
inhibition	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
partial	NN	O	O
CN	NN	O	B-protein
inhibition	NN	O	O
on	NN	O	O
the	NN	O	O
downstream	NN	O	O
signaling	NN	O	O
events	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
gene	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
CsA-treated	NN	O	B-cell_line
leukocytes	NN	O	I-cell_line
stimulated	NN	O	O
by	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
,	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
reduction	NN	O	O
in	NN	O	O
CN	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
similar	NN	O	O
degree	NN	O	O
of	NN	O	O
inhibition	NN	O	O
of	NN	O	O
each	NN	O	O
event	NN	O	O
tested	NN	O	O
:	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
nuclear	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
transfected	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
construct	NN	O	I-DNA
,	NN	O	O
IFN-gamma	NN	O	O
and	NN	O	O
IL-2	NN	O	O
mRNA	NN	O	O
accumulation	NN	O	O
,	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
CN	NN	O	B-protein
inhibition	NN	O	O
was	NN	O	O
reflected	NN	O	O
by	NN	O	O
a	NN	O	O
similar	NN	O	O
degree	NN	O	O
of	NN	O	O
reduction	NN	O	O
in	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
in	NN	O	O
the	NN	O	O
allogeneic	NN	O	B-cell_line
mixed	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
conclusion	NN	O	O
that	NN	O	O
CN	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
rate-limiting	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
reduction	NN	O	O
of	NN	O	O
CN	NN	O	B-protein
activity	NN	O	O
observed	NN	O	O
in	NN	O	O
CsA-treated	NN	O	O
patients	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
similar	NN	O	O
degree	NN	O	O
of	NN	O	O
reduction	NN	O	O
in	NN	O	O
lymphocyte	NN	O	B-DNA
gene	NN	O	I-DNA
activation	NN	O	O
,	NN	O	O
and	NN	O	O
accounts	NN	O	O
for	NN	O	O
the	NN	O	O
immunosuppression	NN	O	O
observed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Selection	NN	O	O
of	NN	O	O
down-regulated	NN	O	B-DNA
sequences	NN	O	I-DNA
along	NN	O	O
the	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
leukemic	NN	O	B-cell_line
HL60	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
dissect	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
we	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
subtractive	NN	O	O
hybridisation	NN	O	O
method	NN	O	O
based	NN	O	O
on	NN	O	O
a	NN	O	O
simplified	NN	O	O
'representational	NN	O	O
difference	NN	O	O
analysis'	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
selected	NN	O	O
16	NN	O	B-DNA
sequences	NN	O	I-DNA
and	NN	O	O
confirmed	NN	O	O
their	NN	O	O
down-regulation	NN	O	O
along	NN	O	O
the	NN	O	O
TPA-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Among	NN	O	O
these	NN	O	O
sequences	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
the	NN	O	O
alpha-tubulin	NN	O	B-protein
,	NN	O	O
the	NN	O	O
TaxREB	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
two	NN	O	O
ribosomal	NN	O	B-DNA
protein	NN	O	I-DNA
sequences	NN	O	I-DNA
which	NN	O	O
had	NN	O	O
not	NN	O	O
been	NN	O	O
previously	NN	O	O
described	NN	O	O
as	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
add	NN	O	O
to	NN	O	O
our	NN	O	O
knowledge	NN	O	O
about	NN	O	O
the	NN	O	O
molecules	NN	O	O
implicated	NN	O	O
along	NN	O	O
the	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
and	NN	O	O
growth	NN	O	O
arrest	NN	O	O
of	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
provide	NN	O	O
a	NN	O	O
first	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
their	NN	O	O
respective	NN	O	O
roles	NN	O	O
.	NN	O	O

-DOCSTART-	O

Control	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
by	NN	O	O
the	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
inhibitor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
maintained	NN	O	O
in	NN	O	O
an	NN	O	O
inactive	NN	O	O
cytoplasmic	NN	O	O
state	NN	O	O
by	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
inhibitors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
proteins	NN	O	I-protein
have	NN	O	O
been	NN	O	O
purified	NN	O	O
and	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
through	NN	O	O
their	NN	O	O
association	NN	O	O
with	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
or	NN	O	I-protein
c-Rel	NN	O	I-protein
subunits	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
isolated	NN	O	O
a	NN	O	O
third	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibitor	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
epsilon	NN	O	I-protein
(	NN	O	O
1	NN	O	O
)	NN	O	O
.	NN	O	O

Upon	NN	O	O
treatment	NN	O	O
with	NN	O	O
a	NN	O	O
large	NN	O	O
variety	NN	O	O
of	NN	O	O
inducers	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
are	NN	O	O
proteolytically	NN	O	O
degraded	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
E29.1	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
hybridoma	NN	O	I-cell_line
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
are	NN	O	O
equally	NN	O	O
associated	NN	O	O
with	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
that	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
is	NN	O	O
degraded	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
what	NN	O	O
has	NN	O	O
been	NN	O	O
originally	NN	O	O
published	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
also	NN	O	O
suggest	NN	O	O
that	NN	O	O
,	NN	O	O
unlike	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
is	NN	O	O
constitutively	NN	O	O
phosphorylated	NN	O	O
and	NN	O	O
resynthesized	NN	O	O
as	NN	O	O
a	NN	O	O
hypophosphorylated	NN	O	B-protein
form	NN	O	I-protein
.	NN	O	O

The	NN	O	O
absence	NN	O	O
of	NN	O	O
slow	NN	O	O
migrating	NN	O	O
forms	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
following	NN	O	O
stimulation	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
does	NN	O	O
not	NN	O	O
necessarily	NN	O	O
constitute	NN	O	O
the	NN	O	O
signal-induced	NN	O	O
event	NN	O	O
which	NN	O	O
targets	NN	O	O
the	NN	O	O
molecule	NN	O	O
for	NN	O	O
proteolysis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Agonistic	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
CD40-specific	NN	O	B-protein
single-chain	NN	O	I-protein
Fv	NN	O	I-protein
constructed	NN	O	O
from	NN	O	O
the	NN	O	O
variable	NN	O	B-protein
regions	NN	O	I-protein
of	NN	O	O
mAb	NN	O	B-protein
G28-5	NN	O	I-protein
.	NN	O	O

A	NN	O	O
single-chain	NN	O	B-protein
Fv	NN	O	I-protein
(	NN	O	O
sFv	NN	O	B-protein
)	NN	O	O
was	NN	O	O
expressed	NN	O	O
from	NN	O	O
the	NN	O	O
variable	NN	O	B-protein
regions	NN	O	I-protein
of	NN	O	O
the	NN	O	O
CD40-specific	NN	O	O
mAb	NN	O	B-protein
G28-5	NN	O	I-protein
.	NN	O	O

The	NN	O	O
molecule	NN	O	O
bound	NN	O	O
CD40	NN	O	B-protein
with	NN	O	O
a	NN	O	O
high	NN	O	O
affinity	NN	O	O
(	NN	O	O
2.2	NN	O	O
nM	NN	O	O
)	NN	O	O
and	NN	O	O
was	NN	O	O
a	NN	O	O
monomer	NN	O	O
in	NN	O	O
solution	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
G28-5	NN	O	B-protein
sFv	NN	O	I-protein
was	NN	O	O
a	NN	O	O
potent	NN	O	O
CD40	NN	O	B-protein
agonist	NN	O	I-protein
that	NN	O	O
rapidly	NN	O	O
crosslinked	NN	O	O
CD40	NN	O	B-protein
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
crosslink	NN	O	O
CD40-Ig	NN	O	B-protein
in	NN	O	O
solution	NN	O	O
.	NN	O	O

G28-5	NN	O	B-protein
sFv	NN	O	I-protein
was	NN	O	O
a	NN	O	O
more	NN	O	O
potent	NN	O	O
agonist	NN	O	O
than	NN	O	O
G28-5	NN	O	B-protein
IgG	NN	O	I-protein
and	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
stimulate	NN	O	O
CD40	NN	O	B-protein
responses	NN	O	O
by	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	O
.	NN	O	O

G28-5	NN	O	B-protein
IgG	NN	O	I-protein
partially	NN	O	O
blocked	NN	O	O
,	NN	O	O
whereas	NN	O	O
G28-5	NN	O	B-protein
sFv	NN	O	I-protein
augmented	NN	O	O
CD40	NN	O	B-protein
responses	NN	O	O
during	NN	O	O
stimulation	NN	O	O
with	NN	O	O
natural	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
gp39-CD8	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
functional	NN	O	O
activity	NN	O	O
of	NN	O	O
ligands	NN	O	O
built	NN	O	O
from	NN	O	O
the	NN	O	O
binding	NN	O	B-protein
site	NN	O	I-protein
of	NN	O	O
G28-5	NN	O	B-protein
is	NN	O	O
highly	NN	O	O
dependent	NN	O	O
upon	NN	O	O
the	NN	O	O
size	NN	O	O
and	NN	O	O
physical	NN	O	O
properties	NN	O	O
of	NN	O	O
the	NN	O	O
molecule	NN	O	O
both	NN	O	O
in	NN	O	O
solution	NN	O	O
and	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surfaces	NN	O	O
.	NN	O	O

-DOCSTART-	O

C/EBP	NN	O	B-protein
activates	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
corticotropin-releasing	NN	O	I-DNA
hormone	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
purpose	NN	O	O
of	NN	O	O
these	NN	O	O
studies	NN	O	O
was	NN	O	O
to	NN	O	O
identify	NN	O	O
whether	NN	O	O
transcription	NN	O	O
factors	NN	O	O
,	NN	O	O
associated	NN	O	O
with	NN	O	O
cytokine	NN	O	O
signalling	NN	O	O
,	NN	O	O
affected	NN	O	O
promoter	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
corticotropin	NN	O	B-DNA
releasing	NN	O	I-DNA
hormone	NN	O	I-DNA
(	NN	O	I-DNA
CRH	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Fragments	NN	O	O
of	NN	O	O
a	NN	O	O
3.6	NN	O	O
kb	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CRH	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
were	NN	O	O
amplified	NN	O	O
by	NN	O	O
PCR	NN	O	O
and	NN	O	O
ligated	NN	O	O
upstream	NN	O	O
of	NN	O	O
a	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
constructs	NN	O	O
were	NN	O	O
transfected	NN	O	O
into	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
lines	NN	O	O
,	NN	O	O
either	NN	O	O
alone	NN	O	O
or	NN	O	O
together	NN	O	O
,	NN	O	O
with	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
expression	NN	O	O
vectors	NN	O	O
.	NN	O	O

Basal	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
3070	NN	O	B-DNA
bp	NN	O	I-DNA
CRH	NN	O	I-DNA
promoter	NN	O	I-DNA
fragment	NN	O	I-DNA
was	NN	O	O
only	NN	O	O
seen	NN	O	O
in	NN	O	O
neuronal	NN	O	O
and	NN	O	O
lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Promoter	NN	O	O
activity	NN	O	O
was	NN	O	O
increased	NN	O	O
by	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
C/EBPbeta	NN	O	B-protein
(	NN	O	O
NF-IL6	NN	O	B-protein
)	NN	O	O
and	NN	O	O
more	NN	O	O
strongly	NN	O	O
,	NN	O	O
by	NN	O	O
C/EBPdelta	NN	O	B-protein
(	NN	O	O
NF-IL6beta	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Increased	NN	O	O
CRH	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
following	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
treatment	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
NF-IL6	NN	O	B-protein
mutant	NN	O	I-protein
,	NN	O	O
showing	NN	O	O
that	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
were	NN	O	O
principally	NN	O	O
mediated	NN	O	O
by	NN	O	O
C/EBP	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
inverse	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CRH	NN	O	B-protein
in	NN	O	O
the	NN	O	O
hypothalamus	NN	O	O
and	NN	O	O
spleens	NN	O	O
of	NN	O	O
arthritic	NN	O	O
rats	NN	O	O
were	NN	O	O
paralleled	NN	O	O
by	NN	O	O
similar	NN	O	O
inverse	NN	O	O
changes	NN	O	O
in	NN	O	O
NF-IL6beta	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
these	NN	O	O
organs	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
some	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
associated	NN	O	O
with	NN	O	O
cytokine	NN	O	B-protein
signalling	NN	O	O
can	NN	O	O
also	NN	O	O
activate	NN	O	O
the	NN	O	O
CRH	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Aberrant	NN	O	O
splicing	NN	O	O
of	NN	O	O
the	NN	O	O
TSG101	NN	O	B-DNA
and	NN	O	I-DNA
FHIT	NN	O	I-DNA
genes	NN	O	I-DNA
occurs	NN	O	O
frequently	NN	O	O
in	NN	O	O
multiple	NN	O	O
malignancies	NN	O	O
and	NN	O	O
in	NN	O	O
normal	NN	O	O
tissues	NN	O	O
and	NN	O	O
mimics	NN	O	O
alterations	NN	O	O
previously	NN	O	O
described	NN	O	O
in	NN	O	O
tumours	NN	O	O
.	NN	O	O

Intragenic	NN	O	O
deletions	NN	O	O
of	NN	O	O
TSG101	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
human	NN	O	O
homolog	NN	O	O
of	NN	O	O
a	NN	O	O
mouse	NN	O	B-DNA
gene	NN	O	I-DNA
(	NN	O	O
tsg101	NN	O	B-DNA
)	NN	O	O
that	NN	O	O
acts	NN	O	O
to	NN	O	O
suppress	NN	O	O
malignant	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
were	NN	O	O
reported	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
breast	NN	O	I-cell_type
tumours	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
screened	NN	O	O
TSG101	NN	O	B-DNA
for	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
in	NN	O	O
DNA	NN	O	O
and	NN	O	O
RNA	NN	O	O
samples	NN	O	O
isolated	NN	O	O
from	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
common	NN	O	O
human	NN	O	O
malignancies	NN	O	O
,	NN	O	O
EBV-immortalised	NN	O	B-cell_type
B-cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
normal	NN	O	O
lung	NN	O	O
parenchyma	NN	O	O
.	NN	O	O

Intragenic	NN	O	O
TSG101	NN	O	B-DNA
deletions	NN	O	O
in	NN	O	O
RNA	NN	O	B-RNA
transcripts	NN	O	I-RNA
were	NN	O	O
frequently	NN	O	O
found	NN	O	O
in	NN	O	O
all	NN	O	O
types	NN	O	O
of	NN	O	O
samples	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
DNA	NN	O	O
failed	NN	O	O
to	NN	O	O
show	NN	O	O
genomic	NN	O	O
rearrangements	NN	O	O
corresponding	NN	O	O
to	NN	O	O
transcripts	NN	O	O
containing	NN	O	O
deletions	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
samples	NN	O	O
.	NN	O	O

The	NN	O	O
breakpoints	NN	O	O
of	NN	O	O
most	NN	O	O
transcript	NN	O	O
deletions	NN	O	O
coincide	NN	O	O
with	NN	O	O
genuine	NN	O	O
or	NN	O	O
cryptic	NN	O	O
splice	NN	O	B-DNA
site	NN	O	I-DNA
sequences	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
they	NN	O	O
result	NN	O	O
from	NN	O	O
alternative	NN	O	O
or	NN	O	O
aberrant	NN	O	O
splicing	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
spectrum	NN	O	O
of	NN	O	O
transcript	NN	O	O
deletions	NN	O	O
has	NN	O	O
previously	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
the	NN	O	O
putative	NN	O	B-DNA
tumour	NN	O	I-DNA
suppressor	NN	O	I-DNA
gene	NN	O	I-DNA
FHIT	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
analysed	NN	O	O
FHIT	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
same	NN	O	O
series	NN	O	O
of	NN	O	O
RNA	NN	O	B-RNA
samples	NN	O	I-RNA
and	NN	O	O
detected	NN	O	O
truncated	NN	O	O
FHIT	NN	O	B-RNA
transcripts	NN	O	I-RNA
frequently	NN	O	O
in	NN	O	O
both	NN	O	O
tumour	NN	O	O
and	NN	O	O
normal	NN	O	O
tissues	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
transcripts	NN	O	O
from	NN	O	O
TSG101	NN	O	B-DNA
,	NN	O	O
FHIT	NN	O	B-DNA
and	NN	O	O
seven	NN	O	O
other	NN	O	O
genes	NN	O	O
were	NN	O	O
analysed	NN	O	O
in	NN	O	O
RNA	NN	O	O
isolated	NN	O	O
from	NN	O	O
normal	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Large	NN	O	O
TSG101	NN	O	B-DNA
and	NN	O	O
FHIT	NN	O	B-DNA
intragenic	NN	O	O
transcript	NN	O	O
deletions	NN	O	O
were	NN	O	O
detected	NN	O	O
and	NN	O	O
these	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
predominant	NN	O	O
transcript	NN	O	O
in	NN	O	O
'aged	NN	O	B-cell_type
'	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Similar	NN	O	O
alterations	NN	O	O
were	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
transcripts	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
genes	NN	O	O
which	NN	O	O
were	NN	O	O
analysed	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
truncated	NN	O	B-RNA
TSG101	NN	O	I-RNA
and	NN	O	I-RNA
FHIT	NN	O	I-RNA
transcripts	NN	O	I-RNA
are	NN	O	O
commonly	NN	O	O
detected	NN	O	O
in	NN	O	O
both	NN	O	O
normal	NN	O	O
and	NN	O	O
malignant	NN	O	O
tissues	NN	O	O
and	NN	O	O
that	NN	O	O
a	NN	O	O
significant	NN	O	O
fraction	NN	O	O
of	NN	O	O
these	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
aberrant	NN	O	O
splicing	NN	O	O
.	NN	O	O

While	NN	O	O
we	NN	O	O
can	NN	O	O
not	NN	O	O
exclude	NN	O	O
that	NN	O	O
alterations	NN	O	O
in	NN	O	O
TSG101	NN	O	B-DNA
and	NN	O	O
FHIT	NN	O	B-DNA
occur	NN	O	O
during	NN	O	O
cancer	NN	O	O
development	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
in	NN	O	O
this	NN	O	O
context	NN	O	O
the	NN	O	O
commonly	NN	O	O
observed	NN	O	O
transcript	NN	O	O
abnormalities	NN	O	O
are	NN	O	O
misleading	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
proliferation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
and	NN	O	I-cell_type
rat	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
curcumin	NN	O	O
,	NN	O	O
a	NN	O	O
curry	NN	O	O
pigment	NN	O	O
.	NN	O	O

Curcumin	NN	O	O
(	NN	O	O
diferuoylmethane	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
yellow	NN	O	O
pigment	NN	O	O
in	NN	O	O
the	NN	O	O
rhizome	NN	O	O
of	NN	O	O
tumeric	NN	O	O
(	NN	O	O
Curcuma	NN	O	O
longa	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
ingredient	NN	O	O
of	NN	O	O
curry	NN	O	O
spice	NN	O	O
,	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
exhibit	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
pharmacological	NN	O	O
effects	NN	O	O
including	NN	O	O
antitumor	NN	O	O
,	NN	O	O
antiinflammatory	NN	O	O
,	NN	O	O
and	NN	O	O
antiinfectious	NN	O	O
activities	NN	O	O
.	NN	O	O

Although	NN	O	O
its	NN	O	O
precise	NN	O	O
mode	NN	O	O
of	NN	O	O
action	NN	O	O
remains	NN	O	O
elusive	NN	O	O
,	NN	O	O
curcumin	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
suppress	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
cells	NN	O	O
stimulated	NN	O	O
to	NN	O	O
proliferate	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
curcumin	NN	O	O
(	NN	O	O
50	NN	O	O
microM	NN	O	O
)	NN	O	O
inhibited	NN	O	O
proliferation	NN	O	O
of	NN	O	O
rat	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
concanavalin	NN	O	O
A	NN	O	O
(	NN	O	O
Con	NN	O	O
A	NN	O	O
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
that	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
Jurkat	NN	O	I-cell_type
lymphoblastoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
logarithmic	NN	O	O
growth	NN	O	O
phase	NN	O	O
.	NN	O	O

The	NN	O	O
pigment	NN	O	O
also	NN	O	O
inhibited	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
dexamethasone-treated	NN	O	B-cell_line
rat	NN	O	I-cell_line
thymocytes	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
UV-irradiated	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
judged	NN	O	O
by	NN	O	O
DNA	NN	O	O
ladder	NN	O	O
formation	NN	O	O
,	NN	O	O
cellular	NN	O	O
morphological	NN	O	O
changes	NN	O	O
,	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
curcumin	NN	O	O
in	NN	O	O
rat	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
partial	NN	O	O
suppression	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Complete	NN	O	O
suppression	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
Con	NN	O	B-cell_line
A-treated	NN	O	I-cell_line
,	NN	O	I-cell_line
proliferating	NN	O	I-cell_line
thymocytes	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
capacity	NN	O	O
of	NN	O	O
curcumin	NN	O	O
to	NN	O	O
inhibit	NN	O	O
both	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
death	NN	O	O
strongly	NN	O	O
implies	NN	O	O
that	NN	O	O
these	NN	O	O
two	NN	O	O
biological	NN	O	O
processes	NN	O	O
share	NN	O	O
a	NN	O	O
common	NN	O	O
pathway	NN	O	O
at	NN	O	O
some	NN	O	O
point	NN	O	O
and	NN	O	O
that	NN	O	O
curcumin	NN	O	O
affects	NN	O	O
a	NN	O	O
common	NN	O	O
step	NN	O	O
,	NN	O	O
presumably	NN	O	O
involving	NN	O	O
a	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Interleukin-7	NN	O	B-protein
upregulates	NN	O	O
the	NN	O	O
interleukin-2-gene	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
by	NN	O	O
enhancing	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activities	NN	O	O
of	NN	O	O
both	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
stromal-derived	NN	O	B-protein
cytokine	NN	O	I-protein
interleukin-7	NN	O	I-protein
(	NN	O	O
IL-7	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2-gene	NN	O	B-DNA
regulation	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Production	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
requires	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2-gene	NN	O	B-DNA
regulation	NN	O	O
.	NN	O	O

T-cell	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	B-protein
TCR	NN	O	I-protein
)	NN	O	I-protein
/CD3	NN	O	I-protein
engagement	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
,	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NFkappaB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
responsive	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	O
CD28RC	NN	O	B-protein
)	NN	O	O
is	NN	O	O
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
signal	NN	O	O
.	NN	O	O

Costimulation	NN	O	O
of	NN	O	O
phytohemagglutinin/anti-CD28	NN	O	B-cell_line
activated	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
with	NN	O	O
IL-7	NN	O	B-protein
induces	NN	O	O
a	NN	O	O
fivefold	NN	O	O
enhanced	NN	O	O
IL-2	NN	O	B-protein
-mRNA	NN	O	O
accumulation	NN	O	O
and	NN	O	O
a	NN	O	O
2.5-fold	NN	O	O
enhanced	NN	O	O
protein	NN	O	O
secretion	NN	O	O
.	NN	O	O

The	NN	O	O
IL-2-gene	NN	O	B-DNA
transcription	NN	O	O
rate	NN	O	O
is	NN	O	O
increased	NN	O	O
3.4-fold	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
IL-7	NN	O	B-protein
is	NN	O	O
in	NN	O	O
part	NN	O	O
mediated	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
the	NN	O	O
IL-7	NN	O	B-protein
effect	NN	O	O
involve	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
(	NN	O	O
60	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
(	NN	O	O
120	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
without	NN	O	O
affecting	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
and	NN	O	O
CD28RC	NN	O	B-protein
,	NN	O	O
which	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
transfection	NN	O	O
assays	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
IL-7	NN	O	B-protein
-induced	NN	O	O
enhancement	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
-DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
not	NN	O	O
cyclosporin	NN	O	O
A-sensitive	NN	O	O
.	NN	O	O

Since	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
IL-7	NN	O	B-protein
-signaling	NN	O	O
pathway	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
fos	NN	O	B-protein
and	NN	O	I-protein
jun	NN	O	I-protein
proteins	NN	O	I-protein
of	NN	O	O
which	NN	O	O
AP-1	NN	O	B-protein
consists	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
spatial	NN	O	O
distribution	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
genes	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
nucleus	NN	O	O
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
gene	NN	O	O
topography	NN	O	O
independent	NN	O	O
of	NN	O	O
cell	NN	O	O
type	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
three-dimensional	NN	O	O
positioning	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
(	NN	O	I-DNA
Ig	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
investigated	NN	O	O
using	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
and	NN	O	O
confocal	NN	O	O
microscopy	NN	O	O
.	NN	O	O

The	NN	O	O
visualization	NN	O	O
of	NN	O	O
heavy	NN	O	B-DNA
and	NN	O	I-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
B-lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
showed	NN	O	O
that	NN	O	O
the	NN	O	O
three	NN	O	O
Ig	NN	O	B-DNA
genes	NN	O	I-DNA
are	NN	O	O
differentially	NN	O	O
and	NN	O	O
nonrandomly	NN	O	O
distributed	NN	O	O
in	NN	O	O
different	NN	O	O
nuclear	NN	O	O
subvolumes	NN	O	O
:	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
genes	NN	O	I-DNA
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
preferentially	NN	O	O
confined	NN	O	O
to	NN	O	O
an	NN	O	O
outer	NN	O	O
nuclear	NN	O	O
volume	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
and	NN	O	I-DNA
lambda	NN	O	I-DNA
genes	NN	O	I-DNA
consistently	NN	O	O
occupied	NN	O	O
more	NN	O	O
central	NN	O	O
positions	NN	O	O
within	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
the	NN	O	O
lambda	NN	O	B-DNA
genes	NN	O	I-DNA
being	NN	O	O
more	NN	O	O
interior	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
data	NN	O	O
further	NN	O	O
show	NN	O	O
that	NN	O	O
these	NN	O	O
overall	NN	O	O
topographical	NN	O	O
distributions	NN	O	O
are	NN	O	O
independent	NN	O	O
of	NN	O	O
gene	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
and	NN	O	O
are	NN	O	O
conserved	NN	O	O
in	NN	O	O
different	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
subtle	NN	O	O
gene	NN	O	O
movements	NN	O	O
within	NN	O	O
those	NN	O	O
defined	NN	O	O
topographical	NN	O	O
regions	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
excluded	NN	O	O
by	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
tissue	NN	O	O
specificity	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
not	NN	O	O
accompanied	NN	O	O
by	NN	O	O
drastic	NN	O	O
changes	NN	O	O
in	NN	O	O
gene	NN	O	O
nuclear	NN	O	O
topography	NN	O	O
,	NN	O	O
rather	NN	O	O
suggesting	NN	O	O
that	NN	O	O
gene	NN	O	O
organization	NN	O	O
within	NN	O	O
the	NN	O	O
nucleus	NN	O	O
may	NN	O	O
be	NN	O	O
primarily	NN	O	O
dependent	NN	O	O
on	NN	O	O
structural	NN	O	O
constraints	NN	O	O
imposed	NN	O	O
on	NN	O	O
the	NN	O	O
respective	NN	O	O
chromosomes	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Dominant	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
response	NN	O	O
to	NN	O	O
the	NN	O	O
immediate-early	NN	O	B-protein
trans-activator	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
BZLF1	NN	O	B-protein
,	NN	O	O
in	NN	O	O
persistent	NN	O	O
type	NN	O	O
A	NN	O	O
or	NN	O	O
B	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
infection	NN	O	O
.	NN	O	O

Five	NN	O	O
healthy	NN	O	O
human	NN	O	O
leukocyte	NN	O	O
antigen-B8	NN	O	O
(	NN	O	O
HLA-B8	NN	O	O
)	NN	O	O
-positive	NN	O	O
virus	NN	O	O
carriers	NN	O	O
were	NN	O	O
studied	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
CD8+	NN	O	B-cell_type
cytotoxic	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
(	NN	O	O
CTL	NN	O	O
)	NN	O	O
response	NN	O	O
to	NN	O	O
an	NN	O	O
HLA-B8-restricted	NN	O	O
peptide	NN	O	O
,	NN	O	O
RAKFKQLLQ	NN	O	O
,	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
(	NN	O	I-protein
EBV	NN	O	I-protein
)	NN	O	I-protein
immediate-early	NN	O	I-protein
trans-activator	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
BZLF1	NN	O	B-protein
.	NN	O	O

Of	NN	O	O
the	NN	O	O
5	NN	O	O
virus	NN	O	O
carriers	NN	O	O
,	NN	O	O
4	NN	O	O
were	NN	O	O
infected	NN	O	O
with	NN	O	O
type	NN	O	O
A	NN	O	O
and	NN	O	O
1	NN	O	O
with	NN	O	O
type	NN	O	O
B	NN	O	O
EBV	NN	O	O
.	NN	O	O

Using	NN	O	O
limiting-dilution	NN	O	O
analysis	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
high	NN	O	O
RAKFKQLLQ-specific	NN	O	O
CTL	NN	O	O
precursor	NN	O	O
frequency	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
after	NN	O	O
specific	NN	O	O
peptide	NN	O	O
or	NN	O	O
autologous	NN	O	O
lymphoblastoid	NN	O	O
cell	NN	O	O
line	NN	O	O
stimulation	NN	O	O
in	NN	O	O
both	NN	O	O
type	NN	O	O
A	NN	O	O
and	NN	O	O
type	NN	O	O
B	NN	O	O
EBV	NN	O	O
carriers	NN	O	O
.	NN	O	O

The	NN	O	O
RAKFKQLLQ-specific	NN	O	O
CTL	NN	O	O
precursor	NN	O	O
frequencies	NN	O	O
in	NN	O	O
all	NN	O	O
5	NN	O	O
persons	NN	O	O
were	NN	O	O
at	NN	O	O
least	NN	O	O
as	NN	O	O
dominant	NN	O	O
as	NN	O	O
those	NN	O	O
observed	NN	O	O
with	NN	O	O
two	NN	O	O
other	NN	O	O
EBV-associated	NN	O	B-protein
,	NN	O	I-protein
HLA-B8-restricted	NN	O	I-protein
latent	NN	O	I-protein
epitopes	NN	O	I-protein
,	NN	O	O
FLRGRAYGL	NN	O	O
and	NN	O	O
QAKWRLQTL	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
show	NN	O	O
that	NN	O	O
healthy	NN	O	O
virus	NN	O	O
carriers	NN	O	O
maintain	NN	O	O
a	NN	O	O
high	NN	O	O
frequency	NN	O	O
of	NN	O	O
BZLF1-specific	NN	O	B-cell_type
memory	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
potentially	NN	O	O
to	NN	O	O
control	NN	O	O
virus	NN	O	O
spread	NN	O	O
from	NN	O	O
lytically	NN	O	B-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
A-MYB	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
its	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
activity	NN	O	O
and	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
110-kDa	NN	O	I-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
studies	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
transgenic	NN	O	O
animals	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
A-MYB	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
plays	NN	O	O
central	NN	O	O
and	NN	O	O
specific	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
and/or	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
highly	NN	O	O
expressed	NN	O	O
in	NN	O	O
Burkitt	NN	O	B-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
may	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
this	NN	O	O
disease	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
therefore	NN	O	O
investigated	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
A-MYB	NN	O	B-protein
and	NN	O	O
its	NN	O	O
regulation	NN	O	O
in	NN	O	O
several	NN	O	O
human	NN	O	B-cell_line
lymphoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
using	NN	O	O
co-transfection	NN	O	O
assays	NN	O	O
and	NN	O	O
show	NN	O	O
that	NN	O	O
A-MYB	NN	O	B-protein
is	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
in	NN	O	O
all	NN	O	O
the	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
studied	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
particular	NN	O	O
the	NN	O	O
best	NN	O	O
responder	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
was	NN	O	O
the	NN	O	O
Burkitt	NN	O	B-cell_line
's	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Namalwa	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
A-MYB	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
not	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
observed	NN	O	O
when	NN	O	O
either	NN	O	O
an	NN	O	O
artificial	NN	O	O
construct	NN	O	O
or	NN	O	O
the	NN	O	O
c-MYC	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
reporter	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
functional	NN	O	B-protein
domains	NN	O	I-protein
responsible	NN	O	O
for	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
transactivation	NN	O	O
,	NN	O	O
and	NN	O	O
negative	NN	O	O
regulation	NN	O	O
,	NN	O	O
previously	NN	O	O
characterized	NN	O	O
in	NN	O	O
a	NN	O	O
fibroblast	NN	O	O
context	NN	O	O
,	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
similar	NN	O	O
activity	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
region	NN	O	O
of	NN	O	O
A-MYB	NN	O	B-protein
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
B	NN	O	O
cell	NN	O	O
specific	NN	O	O
activity	NN	O	O
was	NN	O	O
defined	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
218	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
containing	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
a	NN	O	O
110-kDa	NN	O	B-protein
protein	NN	O	I-protein
has	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
B	NN	O	B-cell_line
,	NN	O	I-cell_line
but	NN	O	I-cell_line
not	NN	O	I-cell_line
T	NN	O	I-cell_line
,	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
specifically	NN	O	O
binds	NN	O	O
to	NN	O	O
this	NN	O	O
A-MYB	NN	O	B-protein
N-terminal	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

We	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
this	NN	O	O
110-kDa	NN	O	B-protein
protein	NN	O	I-protein
may	NN	O	O
be	NN	O	O
a	NN	O	O
functionally	NN	O	O
important	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
co-activator	NN	O	I-protein
of	NN	O	O
A-MYB	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	O
factor	NN	O	O
Egr-1	NN	O	B-protein
activity	NN	O	O
down-regulates	NN	O	O
Fas	NN	O	O
and	NN	O	O
CD23	NN	O	O
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
mature	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
Ag	NN	O	B-protein
receptor	NN	O	I-protein
cross-linking	NN	O	O
induces	NN	O	O
transient	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Egr-1	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
the	NN	O	O
activating	NN	O	O
signals	NN	O	O
leading	NN	O	O
to	NN	O	O
Egr-1	NN	O	B-protein
induction	NN	O	O
have	NN	O	O
been	NN	O	O
studied	NN	O	O
extensively	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
genes	NN	O	O
that	NN	O	O
are	NN	O	O
placed	NN	O	O
further	NN	O	O
downstream	NN	O	O
within	NN	O	O
this	NN	O	O
activation	NN	O	O
cascade	NN	O	O
and	NN	O	O
that	NN	O	O
are	NN	O	O
transcriptionally	NN	O	O
regulated	NN	O	O
by	NN	O	O
Egr-1	NN	O	B-protein
.	NN	O	O

To	NN	O	O
identify	NN	O	O
such	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
established	NN	O	O
Egr-1-overexpressing	NN	O	B-cell_line
transfectants	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
K46	NN	O	I-cell_line
and	NN	O	O
from	NN	O	O
human	NN	O	B-cell_line
Ramos	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

All	NN	O	O
clones	NN	O	O
derived	NN	O	O
from	NN	O	O
K46	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
showed	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
CD44	NN	O	B-protein
.	NN	O	O

Most	NN	O	O
interestingly	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
CD95	NN	O	B-protein
(	NN	O	O
Fas/Apo-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
of	NN	O	O
CD23	NN	O	B-protein
was	NN	O	O
down-regulated	NN	O	O
in	NN	O	O
all	NN	O	O
K46	NN	O	B-cell_line
transfectants	NN	O	I-cell_line
.	NN	O	O

As	NN	O	O
a	NN	O	O
consequence	NN	O	O
,	NN	O	O
they	NN	O	O
became	NN	O	O
resistant	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
anti-CD95	NN	O	B-protein
Ab	NN	O	I-protein
treatment	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
the	NN	O	O
Egr-1-expressing	NN	O	B-cell_line
Ramos	NN	O	I-cell_line
cells	NN	O	I-cell_line
showed	NN	O	O
reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
CD95	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Egr-1	NN	O	B-protein
seems	NN	O	O
to	NN	O	O
control	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
downstream	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
not	NN	O	O
only	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
as	NN	O	O
a	NN	O	O
repressor	NN	O	B-protein
molecule	NN	O	I-protein
.	NN	O	O

In	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
Egr-1	NN	O	B-protein
therefore	NN	O	O
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
integrating	NN	O	O
the	NN	O	O
short-lived	NN	O	O
signal	NN	O	O
delivered	NN	O	O
by	NN	O	O
triggering	NN	O	O
of	NN	O	O
the	NN	O	O
Ag	NN	O	B-protein
receptor	NN	O	I-protein
into	NN	O	O
phenotypic	NN	O	O
changes	NN	O	O
,	NN	O	O
including	NN	O	O
repression	NN	O	O
of	NN	O	O
CD95	NN	O	B-protein
and	NN	O	O
CD23	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
by	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
alpha	NN	O	O
and	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
beta	NN	O	O
stability	NN	O	O
.	NN	O	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
must	NN	O	O
be	NN	O	O
released	NN	O	O
from	NN	O	O
cytoplasmic	NN	O	B-protein
inhibitory	NN	O	I-protein
molecules	NN	O	I-protein
(	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
Bs	NN	O	I-protein
)	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
move	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
and	NN	O	O
to	NN	O	O
activate	NN	O	O
its	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
regulating	NN	O	O
the	NN	O	O
maintenance	NN	O	O
of	NN	O	O
constitutive	NN	O	B-protein
nuclear	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
some	NN	O	O
cell-types	NN	O	O
and	NN	O	O
of	NN	O	O
sustained	NN	O	O
nuclear	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Increased	NN	O	O
turnover	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
compared	NN	O	O
the	NN	O	O
turnover	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
in	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Both	NN	O	O
proteins	NN	O	O
display	NN	O	O
a	NN	O	O
high	NN	O	O
turnover	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
although	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
is	NN	O	O
considerably	NN	O	O
more	NN	O	O
stable	NN	O	O
than	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
half-life	NN	O	O
of	NN	O	O
both	NN	O	O
inhibitors	NN	O	O
is	NN	O	O
increased	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
all	NN	O	O
other	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
/I	NN	O	O
kappa	NN	O	O
B	NN	O	O
molecules	NN	O	O
tested	NN	O	O
are	NN	O	O
relatively	NN	O	O
stable	NN	O	O
in	NN	O	O
both	NN	O	O
cell-types	NN	O	O
.	NN	O	O

The	NN	O	O
elevated	NN	O	O
turnover	NN	O	O
of	NN	O	O
endogenous	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
in	NN	O	O
Namalwa	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
proteasome	NN	O	O
inhibitor	NN	O	O
and	NN	O	O
thus	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
driven	NN	O	O
by	NN	O	O
the	NN	O	O
same	NN	O	O
degradation	NN	O	O
machinery	NN	O	O
as	NN	O	O
the	NN	O	O
slower	NN	O	O
turnover	NN	O	O
in	NN	O	O
non-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
processes	NN	O	O
involved	NN	O	O
in	NN	O	O
persistent	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

TNF-alpha	NN	O	O
signaling	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
rapid	NN	O	O
depletion	NN	O	O
of	NN	O	O
cellular	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
pools	NN	O	O
.	NN	O	O

I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
is	NN	O	O
efficiently	NN	O	O
resynthesized	NN	O	O
whereas	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
levels	NN	O	O
stay	NN	O	O
low	NN	O	O
for	NN	O	O
a	NN	O	O
prolonged	NN	O	O
time	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
can	NN	O	O
be	NN	O	O
detected	NN	O	O
for	NN	O	O
several	NN	O	O
hours	NN	O	O
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
removal	NN	O	O
of	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-protein
containing	NN	O	O
medium	NN	O	O
causes	NN	O	O
a	NN	O	O
rapid	NN	O	O
decrease	NN	O	O
in	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

A	NN	O	O
phosphoform	NN	O	O
of	NN	O	O
newly	NN	O	O
synthesized	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
is	NN	O	O
visible	NN	O	O
when	NN	O	O
degradation	NN	O	O
by	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
is	NN	O	O
inhibited	NN	O	O
and	NN	O	O
new	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
displays	NN	O	O
the	NN	O	O
same	NN	O	O
properties	NN	O	O
regarding	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
second	NN	O	O
inducer	NN	O	O
.	NN	O	O

There	NN	O	O
is	NN	O	O
no	NN	O	O
significant	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
turnover	NN	O	O
of	NN	O	O
pre-	NN	O	O
and	NN	O	O
post-inductive	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
resynthesis	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
removal	NN	O	O
of	NN	O	O
the	NN	O	O
stimulus	NN	O	O
are	NN	O	O
obligatory	NN	O	O
steps	NN	O	O
for	NN	O	O
the	NN	O	O
inactivation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
NF	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Selection	NN	O	O
of	NN	O	O
a	NN	O	O
diverse	NN	O	O
TCR	NN	O	B-protein
repertoire	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
an	NN	O	O
Epstein-Barr	NN	O	B-protein
virus-encoded	NN	O	I-protein
transactivator	NN	O	I-protein
protein	NN	O	I-protein
BZLF1	NN	O	B-protein
by	NN	O	O
CD8+	NN	O	B-cell_type
cytotoxic	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
during	NN	O	O
primary	NN	O	O
and	NN	O	O
persistent	NN	O	O
infection	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
CD8+	NN	O	B-protein
cytotoxic	NN	O	I-protein
T	NN	O	I-protein
lymphocyte	NN	O	I-protein
(	NN	O	I-protein
CTL	NN	O	I-protein
)	NN	O	I-protein
repertoire	NN	O	I-protein
to	NN	O	O
an	NN	O	O
HLA	NN	O	O
B8-restricted	NN	O	O
peptide	NN	O	O
,	NN	O	O
RAKFKQLLQ	NN	O	O
,	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
(	NN	O	I-protein
EBV	NN	O	I-protein
)	NN	O	I-protein
immediate-early	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
BZLF1	NN	O	B-protein
.	NN	O	O

Repertoire	NN	O	O
selection	NN	O	O
was	NN	O	O
monitored	NN	O	O
by	NN	O	O
determining	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
beta	NN	O	I-protein
chain	NN	O	I-protein
sequences	NN	O	I-protein
of	NN	O	O
RAKFKQLLQ-specific	NN	O	B-cell_line
CTL	NN	O	I-cell_line
established	NN	O	O
from	NN	O	O
primary	NN	O	O
infected	NN	O	O
and	NN	O	O
healthy	NN	O	O
virus	NN	O	O
carriers	NN	O	O
.	NN	O	O

PCR	NN	O	O
analysis	NN	O	O
of	NN	O	O
spontaneous	NN	O	B-cell_line
EBV-transformed	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
LCL	NN	O	B-cell_line
)	NN	O	O
from	NN	O	O
three	NN	O	O
individuals	NN	O	O
with	NN	O	O
primary	NN	O	O
infection	NN	O	O
showed	NN	O	O
that	NN	O	O
two	NN	O	O
were	NN	O	O
infected	NN	O	O
with	NN	O	O
type	NN	O	O
A	NN	O	O
and	NN	O	O
one	NN	O	O
with	NN	O	O
type	NN	O	O
B	NN	O	O
EBV	NN	O	O
.	NN	O	O

Polyclonal	NN	O	B-cell_line
and	NN	O	I-cell_line
clonal	NN	O	I-cell_line
CTL	NN	O	I-cell_line
that	NN	O	O
were	NN	O	O
generated	NN	O	O
by	NN	O	O
stimulating	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
an	NN	O	O
HLA	NN	O	B-cell_line
B8+	NN	O	I-cell_line
homozygous	NN	O	I-cell_line
LCL	NN	O	I-cell_line
lysed	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
blasts	NN	O	I-cell_line
pulsed	NN	O	O
with	NN	O	O
the	NN	O	O
peptide	NN	O	O
,	NN	O	O
RAKFKQLLQ	NN	O	O
;	NN	O	O
lysis	NN	O	O
of	NN	O	O
certain	NN	O	O
HLA	NN	O	O
B8+	NN	O	O
LCL	NN	O	O
targets	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
abundance	NN	O	O
of	NN	O	O
BZLF1	NN	O	B-RNA
transcripts	NN	O	I-RNA
.	NN	O	O

TCR	NN	O	B-DNA
beta	NN	O	I-DNA
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
while	NN	O	O
there	NN	O	O
was	NN	O	O
loop	NN	O	O
length	NN	O	O
restriction	NN	O	O
in	NN	O	O
the	NN	O	O
putative	NN	O	O
peptide	NN	O	O
contact	NN	O	O
site	NN	O	O
of	NN	O	O
all	NN	O	O
responding	NN	O	O
beta	NN	O	B-protein
chains	NN	O	I-protein
,	NN	O	O
diverse	NN	O	O
and	NN	O	O
unique	NN	O	B-cell_line
(	NN	O	I-cell_line
non-recurrent	NN	O	I-cell_line
)	NN	O	I-cell_line
TCR	NN	O	I-cell_line
beta	NN	O	I-cell_line
clonotypes	NN	O	I-cell_line
were	NN	O	O
selected	NN	O	O
in	NN	O	O
individuals	NN	O	O
during	NN	O	O
primary	NN	O	O
infection	NN	O	O
and	NN	O	O
continued	NN	O	O
to	NN	O	O
emerge	NN	O	O
after	NN	O	O
long-term	NN	O	O
virus	NN	O	O
exposure	NN	O	O
.	NN	O	O

TCR	NN	O	B-protein
-contact	NN	O	O
site	NN	O	O
heterogeneity	NN	O	O
was	NN	O	O
excluded	NN	O	O
as	NN	O	O
the	NN	O	O
selective	NN	O	O
force	NN	O	O
in	NN	O	O
diversity	NN	O	O
generation	NN	O	O
since	NN	O	O
the	NN	O	O
epitope-encoded	NN	O	O
sequences	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
identical	NN	O	O
within	NN	O	O
endogenous	NN	O	O
virus	NN	O	O
isolates	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
first	NN	O	O
study	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
repertoire	NN	O	I-protein
selection	NN	O	O
for	NN	O	O
an	NN	O	O
EBV	NN	O	B-protein
lytic	NN	O	I-protein
antigen	NN	O	I-protein
,	NN	O	O
a	NN	O	O
BZLF1	NN	O	B-protein
-reactive	NN	O	O
component	NN	O	O
of	NN	O	O
diverse	NN	O	O
clonotypes	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
primary	NN	O	O
type	NN	O	O
A	NN	O	O
or	NN	O	O
type	NN	O	O
B	NN	O	O
EBV	NN	O	O
infection	NN	O	O
which	NN	O	O
was	NN	O	O
sustained	NN	O	O
in	NN	O	O
the	NN	O	O
EBV-specific	NN	O	O
memory	NN	O	O
response	NN	O	O
throughout	NN	O	O
life-long	NN	O	O
infection	NN	O	O
.	NN	O	O

This	NN	O	O
diversity	NN	O	O
selection	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
maintaining	NN	O	O
a	NN	O	O
balanced	NN	O	O
viral	NN	O	O
load	NN	O	O
throughout	NN	O	O
EBV	NN	O	O
persistence	NN	O	O
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
anoxia/reoxygenation-induced	NN	O	O
neutrophil	NN	O	O
adherence	NN	O	O
to	NN	O	O
cultured	NN	O	B-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
objectives	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
were	NN	O	O
to	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
determine	NN	O	O
the	NN	O	O
time	NN	O	O
course	NN	O	O
of	NN	O	O
neutrophil	NN	O	O
adhesion	NN	O	O
to	NN	O	O
monolayers	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
umbilical	NN	O	I-cell_line
vein	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
HUVECs	NN	O	B-cell_line
)	NN	O	O
that	NN	O	O
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
60	NN	O	O
minutes	NN	O	O
of	NN	O	O
anoxia	NN	O	O
followed	NN	O	O
by	NN	O	O
30	NN	O	O
to	NN	O	O
600	NN	O	O
minutes	NN	O	O
of	NN	O	O
reoxygenation	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
define	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
responsible	NN	O	O
for	NN	O	O
both	NN	O	O
the	NN	O	O
early	NN	O	O
(	NN	O	O
minutes	NN	O	O
)	NN	O	O
and	NN	O	O
late	NN	O	O
(	NN	O	O
hours	NN	O	O
)	NN	O	O
hyperadhesivity	NN	O	O
of	NN	O	O
postanoxic	NN	O	B-cell_line
HUVECs	NN	O	I-cell_line
to	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
results	NN	O	O
clearly	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
anoxia/reoxygenation	NN	O	O
(	NN	O	O
A/R	NN	O	O
)	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
biphasic	NN	O	O
increase	NN	O	O
in	NN	O	O
neutrophil	NN	O	O
adhesion	NN	O	O
to	NN	O	O
HUVECs	NN	O	B-cell_line
,	NN	O	O
with	NN	O	O
peak	NN	O	O
responses	NN	O	O
occurring	NN	O	O
at	NN	O	O
30	NN	O	O
minutes	NN	O	O
(	NN	O	O
phase	NN	O	O
1	NN	O	O
)	NN	O	O
and	NN	O	O
240	NN	O	O
minutes	NN	O	O
(	NN	O	O
phase	NN	O	O
2	NN	O	O
)	NN	O	O
after	NN	O	O
reoxygenation	NN	O	O
.	NN	O	O

Oxypurinol	NN	O	O
and	NN	O	O
catalase	NN	O	B-protein
inhibited	NN	O	O
phase-1	NN	O	O
adhesion	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
xanthine	NN	O	B-protein
oxidase	NN	O	I-protein
and	NN	O	O
H2O2	NN	O	O
.	NN	O	O

In	NN	O	O
comparison	NN	O	O
,	NN	O	O
platelet	NN	O	B-protein
activating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
PAF	NN	O	B-protein
)	NN	O	O
contributed	NN	O	O
to	NN	O	O
both	NN	O	O
phases	NN	O	O
of	NN	O	O
neutrophil	NN	O	O
adhesion	NN	O	O
.	NN	O	O

Anti-intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
anti-P-selectin	NN	O	B-protein
antibodies	NN	O	I-protein
(	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
[	NN	O	O
mAbs	NN	O	B-protein
]	NN	O	O
)	NN	O	O
attenuated	NN	O	O
phase-1	NN	O	O
neutrophil	NN	O	O
adhesion	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
roles	NN	O	O
for	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
ICAM-1	NN	O	B-protein
and	NN	O	O
enhanced	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
preformed	NN	O	O
P-selectin	NN	O	B-protein
.	NN	O	O

Phase-2	NN	O	O
neutrophil	NN	O	O
adhesion	NN	O	O
was	NN	O	O
attenuated	NN	O	O
by	NN	O	O
an	NN	O	O
anti-E-selectin	NN	O	B-protein
mAb	NN	O	I-protein
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
dominant	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
in	NN	O	O
the	NN	O	O
late	NN	O	O
phase	NN	O	O
response	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
with	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
and	NN	O	O
cycloheximide	NN	O	O
or	NN	O	O
with	NN	O	O
competing	NN	O	O
ds-oligonucleotides	NN	O	O
containing	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
or	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
cognate	NN	O	B-DNA
DNA	NN	O	I-DNA
sequences	NN	O	I-DNA
significantly	NN	O	O
attenuated	NN	O	O
phase-2	NN	O	O
response	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
de	NN	O	O
novo	NN	O	O
macromolecule	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Surface	NN	O	O
expression	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
P-selectin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
E-selectin	NN	O	B-protein
on	NN	O	O
HUVECs	NN	O	B-cell_line
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
phase-1	NN	O	O
and	NN	O	O
-2	NN	O	O
neutrophil	NN	O	O
adhesion	NN	O	O
responses	NN	O	O
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
A/R	NN	O	O
elicits	NN	O	O
a	NN	O	O
two-phase	NN	O	O
neutrophil-endothelial	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
response	NN	O	O
that	NN	O	O
involves	NN	O	O
transcription-independent	NN	O	O
and	NN	O	O
transcription-dependent	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
different	NN	O	O
endothelial	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
determinants	NN	O	O
regulating	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

CD40	NN	O	B-protein
signaling	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
important	NN	O	O
for	NN	O	O
generating	NN	O	O
an	NN	O	O
effective	NN	O	O
humoral	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

CD40	NN	O	B-protein
ligation	NN	O	O
leads	NN	O	O
to	NN	O	O
B	NN	O	O
cell	NN	O	O
activation	NN	O	O
events	NN	O	O
such	NN	O	O
as	NN	O	O
proliferation	NN	O	O
,	NN	O	O
Ig	NN	O	O
secretion	NN	O	O
,	NN	O	O
isotype	NN	O	O
switching	NN	O	O
,	NN	O	O
and	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
memory	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Many	NN	O	O
of	NN	O	O
these	NN	O	O
events	NN	O	O
are	NN	O	O
dependent	NN	O	O
upon	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

To	NN	O	O
define	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
components	NN	O	O
upstream	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
the	NN	O	O
functional	NN	O	O
consequences	NN	O	O
downstream	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
mouse	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
transfectants	NN	O	I-cell_line
expressing	NN	O	O
wild-type	NN	O	B-protein
or	NN	O	I-protein
mutant	NN	O	I-protein
human	NN	O	I-protein
CD40	NN	O	I-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
truncation	NN	O	O
and	NN	O	O
point	NN	O	O
mutants	NN	O	O
defined	NN	O	O
a	NN	O	O
10-amino	NN	O	B-protein
acid	NN	O	I-protein
CD40	NN	O	I-protein
cytoplasmic	NN	O	I-protein
signaling	NN	O	I-protein
determinant	NN	O	I-protein
required	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

A	NN	O	O
threonine	NN	O	O
residue	NN	O	O
at	NN	O	O
position	NN	O	B-protein
234	NN	O	I-protein
,	NN	O	O
previously	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
CD40	NN	O	B-protein
association	NN	O	O
with	NN	O	O
TNF	NN	O	B-protein
receptor-associated	NN	O	I-protein
factor	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	O
TRAF2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
TRAF3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TRAF5	NN	O	B-protein
,	NN	O	O
was	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
CD40	NN	O	B-protein
-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
can	NN	O	O
occur	NN	O	O
independently	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
and	NN	O	O
TRAF5	NN	O	B-protein
association	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
transmembrane	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
CD40	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
p23	NN	O	B-protein
,	NN	O	O
a	NN	O	O
molecule	NN	O	O
that	NN	O	O
associates	NN	O	O
with	NN	O	O
CD40	NN	O	B-protein
in	NN	O	O
a	NN	O	O
region	NN	O	O
other	NN	O	O
than	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

Proteasome-dependent	NN	O	O
inhibitory	NN	O	O
kappa	NN	O	O
B	NN	O	O
alpha	NN	O	O
(	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	O
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
degradation	NN	O	O
occurred	NN	O	O
downstream	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
ligation	NN	O	O
and	NN	O	O
preceded	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
nuclear	NN	O	O
translocation	NN	O	O
.	NN	O	O

CD40-	NN	O	O
or	NN	O	O
pervanadate-mediated	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
was	NN	O	O
not	NN	O	O
detected	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
Ab	NN	O	O
secretion	NN	O	O
and	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
and	NN	O	O
LFA-1	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
insufficient	NN	O	O
for	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
B7-1	NN	O	B-protein
,	NN	O	O
Fas	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CD23	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

CD30	NN	O	B-protein
-dependent	NN	O	O
degradation	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
:	NN	O	O
implications	NN	O	O
for	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
TRAF	NN	O	B-protein
signaling	NN	O	O
and	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
cell	NN	O	O
survival	NN	O	O
.	NN	O	O

CD30	NN	O	B-protein
is	NN	O	O
a	NN	O	O
cell-surface	NN	O	B-protein
receptor	NN	O	I-protein
that	NN	O	O
can	NN	O	O
augment	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
and	NN	O	O
survival	NN	O	O
through	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

CD30	NN	O	B-protein
,	NN	O	O
however	NN	O	O
,	NN	O	O
has	NN	O	O
also	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptotic	NN	O	O
cell	NN	O	O
death	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
CD30	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
is	NN	O	O
to	NN	O	O
render	NN	O	O
cells	NN	O	O
sensitive	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
type	NN	O	B-protein
1	NN	O	I-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TNFR1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

This	NN	O	O
sensitization	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
TRAF-binding	NN	O	B-protein
sites	NN	O	I-protein
within	NN	O	O
the	NN	O	O
CD30	NN	O	B-protein
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
proteins	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
these	NN	O	O
sites	NN	O	O
is	NN	O	O
TRAF2	NN	O	B-protein
,	NN	O	O
a	NN	O	O
signal	NN	O	B-protein
transduction	NN	O	I-protein
molecule	NN	O	I-protein
that	NN	O	O
is	NN	O	O
also	NN	O	O
utilized	NN	O	O
by	NN	O	O
TNFR1	NN	O	B-protein
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
several	NN	O	O
downstream	NN	O	O
kinases	NN	O	B-protein
and	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

During	NN	O	O
CD30	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
to	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
CD30	NN	O	B-protein
results	NN	O	O
in	NN	O	O
the	NN	O	O
rapid	NN	O	O
depletion	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
and	NN	O	O
the	NN	O	O
associated	NN	O	O
protein	NN	O	O
TRAF1	NN	O	B-protein
by	NN	O	O
proteolysis	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
CD30	NN	O	B-protein
limits	NN	O	O
its	NN	O	O
own	NN	O	O
ability	NN	O	O
to	NN	O	O
transduce	NN	O	O
cell	NN	O	O
survival	NN	O	O
signals	NN	O	O
through	NN	O	O
signal-coupled	NN	O	O
depletion	NN	O	O
of	NN	O	O
TRAF2	NN	O	B-protein
.	NN	O	O

Depletion	NN	O	O
of	NN	O	O
intracellular	NN	O	O
TRAF2	NN	O	B-protein
and	NN	O	O
its	NN	O	O
coassociated	NN	O	B-protein
proteins	NN	O	I-protein
also	NN	O	O
increased	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
to	NN	O	O
undergoing	NN	O	O
apoptosis	NN	O	O
during	NN	O	O
activation	NN	O	O
of	NN	O	O
death-inducing	NN	O	B-protein
receptors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
TNFR1	NN	O	B-protein
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
dominant-negative	NN	O	B-protein
form	NN	O	I-protein
of	NN	O	O
TRAF2	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
potentiate	NN	O	O
TNFR1	NN	O	B-protein
-mediated	NN	O	O
death	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
provide	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
through	NN	O	O
which	NN	O	O
CD30	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
other	NN	O	O
TRAF-binding	NN	O	B-protein
members	NN	O	I-protein
of	NN	O	O
the	NN	O	O
TNFR	NN	O	B-protein
superfamily	NN	O	I-protein
,	NN	O	O
can	NN	O	O
negatively	NN	O	O
regulate	NN	O	O
cell	NN	O	O
survival	NN	O	O
.	NN	O	O

-DOCSTART-	O

Analysis	NN	O	O
of	NN	O	O
myeloid-associated	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
distribution	NN	O	O
of	NN	O	O
myeloid	NN	O	B-protein
lineage-associated	NN	O	I-protein
cytokine	NN	O	I-protein
receptors	NN	O	I-protein
and	NN	O	O
lysosomal	NN	O	B-protein
proteins	NN	O	I-protein
was	NN	O	O
analyzed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
CD34+	NN	O	I-cell_line
cord	NN	O	I-cell_line
blood	NN	O	I-cell_line
cell	NN	O	I-cell_line
(	NN	O	I-cell_line
CB	NN	O	I-cell_line
)	NN	O	I-cell_line
subsets	NN	O	I-cell_line
at	NN	O	O
different	NN	O	O
stages	NN	O	O
of	NN	O	O
myeloid	NN	O	O
commitment	NN	O	O
by	NN	O	O
reverse-transcriptase	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
highly	NN	O	O
specific	NN	O	O
granulomonocyte-associated	NN	O	B-protein
lysosomal	NN	O	I-protein
proteins	NN	O	I-protein
myeloperoxidase	NN	O	B-protein
(	NN	O	O
MPO	NN	O	B-protein
)	NN	O	O
and	NN	O	O
lysozyme	NN	O	B-protein
(	NN	O	O
LZ	NN	O	B-protein
)	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
PU.1	NN	O	B-protein
,	NN	O	O
were	NN	O	O
already	NN	O	O
detectable	NN	O	O
in	NN	O	O
the	NN	O	O
most	NN	O	O
immature	NN	O	O
CD34+Thy-1+	NN	O	B-cell_line
subset	NN	O	I-cell_line
.	NN	O	O

Messenger	NN	O	B-RNA
RNA	NN	O	I-RNA
(	NN	O	O
mRNA	NN	O	B-RNA
)	NN	O	O
levels	NN	O	O
for	NN	O	O
the	NN	O	O
granulocyte-colony	NN	O	B-protein
stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
G-CSF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
granulocyte-macrophage	NN	O	B-protein
(	NN	O	I-protein
GM	NN	O	I-protein
)	NN	O	I-protein
-CSF	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
subunit	NN	O	I-protein
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
receptors	NN	O	I-protein
I	NN	O	I-protein
(	NN	O	I-protein
p55	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
II	NN	O	I-protein
(	NN	O	I-protein
p75	NN	O	I-protein
)	NN	O	I-protein
were	NN	O	O
also	NN	O	O
detected	NN	O	O
in	NN	O	O
this	NN	O	O
subset	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
c-kit	NN	O	B-protein
and	NN	O	O
flt-3	NN	O	B-protein
,	NN	O	O
receptors	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
on	NN	O	O
progenitor	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
monocyte-macrophage	NN	O	B-protein
colony	NN	O	I-protein
stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
M-CSF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
was	NN	O	O
largely	NN	O	O
absent	NN	O	O
at	NN	O	O
this	NN	O	O
stage	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
CD34+Thy-1-CD45RA-	NN	O	B-cell_line
subsets	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
M-CSF	NN	O	B-protein
receptor	NN	O	I-protein
was	NN	O	O
first	NN	O	O
detectable	NN	O	O
in	NN	O	O
the	NN	O	O
myeloid-committed	NN	O	B-cell_line
CD34+Thy-l-CD45RA+	NN	O	I-cell_line
subset	NN	O	I-cell_line
.	NN	O	O

All	NN	O	O
other	NN	O	O
molecules	NN	O	O
studied	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
at	NN	O	O
this	NN	O	O
stage	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Different	NN	O	O
cocktails	NN	O	O
of	NN	O	O
the	NN	O	O
identified	NN	O	O
ligands	NN	O	O
were	NN	O	O
added	NN	O	O
to	NN	O	O
sorted	NN	O	B-cell_line
CD34+Thy-1+	NN	O	I-cell_line
single	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Low	NN	O	O
proliferative	NN	O	O
capacity	NN	O	O
was	NN	O	O
observed	NN	O	O
after	NN	O	O
1	NN	O	O
week	NN	O	O
in	NN	O	O
culture	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
SCF	NN	O	B-protein
)	NN	O	O
+	NN	O	O
Flt-3	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
FL	NN	O	B-protein
)	NN	O	O
+	NN	O	O
G-CSF	NN	O	B-protein
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
to	NN	O	O
this	NN	O	O
basic	NN	O	O
cocktail	NN	O	O
consistently	NN	O	O
increased	NN	O	O
the	NN	O	O
clonogenic	NN	O	O
capacity	NN	O	O
of	NN	O	O
single	NN	O	B-cell_line
CD34+Thy-1+	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
this	NN	O	O
effect	NN	O	O
was	NN	O	O
further	NN	O	O
enhanced	NN	O	O
(	NN	O	O
up	NN	O	O
to	NN	O	O
72.3	NN	O	O
+/-	NN	O	O
4.3	NN	O	O
%	NN	O	O
on	NN	O	O
day	NN	O	O
7	NN	O	O
)	NN	O	O
by	NN	O	O
the	NN	O	O
inclusion	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
myeloid-associated	NN	O	B-RNA
growth	NN	O	I-RNA
factor	NN	O	I-RNA
receptor	NN	O	I-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
CD34+	NN	O	B-cell_line
CB	NN	O	I-cell_line
subsets	NN	O	I-cell_line
does	NN	O	O
not	NN	O	O
discriminate	NN	O	O
the	NN	O	O
various	NN	O	O
stages	NN	O	O
of	NN	O	O
differentiation	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
TNF-alpha	NN	O	B-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
costimulatory	NN	O	O
factor	NN	O	O
of	NN	O	O
the	NN	O	O
very	NN	O	O
immature	NN	O	B-cell_line
CD34+Thy-1+	NN	O	I-cell_line
CB	NN	O	I-cell_line
subset	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
endothelial	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphatase	NN	O	O
inhibitors	NN	O	O
:	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
from	NN	O	O
our	NN	O	O
laboratory	NN	O	O
have	NN	O	O
indicated	NN	O	O
that	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
(	NN	O	O
PTPase	NN	O	B-protein
)	NN	O	O
inhibitors	NN	O	O
can	NN	O	O
down-modulate	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
ML-1a	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
Singh	NN	O	O
and	NN	O	O
Aggarwal	NN	O	O
,	NN	O	O
J.	NN	O	O
Biol.	NN	O	O
Chem.	NN	O	O
1995	NN	O	O
:	NN	O	O
270	NN	O	O
:	NN	O	O
10631	NN	O	O
)	NN	O	O
.	NN	O	O

Since	NN	O	O
TNF	NN	O	B-protein
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	O
inducers	NN	O	O
of	NN	O	O
various	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
in	NN	O	O
human	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
and	NN	O	O
their	NN	O	O
expression	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
require	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
PTPase	NN	O	B-protein
inhibitors	NN	O	O
on	NN	O	O
the	NN	O	O
TNF	NN	O	B-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
intracellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
(	NN	O	I-protein
ICAM	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
,	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
(	NN	O	I-protein
VCAM	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
and	NN	O	O
endothelial	NN	O	B-protein
leukocyte	NN	O	I-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
(	NN	O	I-protein
ELAM	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
.	NN	O	O

Like	NN	O	O
ML-1a	NN	O	B-cell_line
,	NN	O	O
human	NN	O	B-cell_line
dermal	NN	O	I-cell_line
microvessel	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
MVEC	NN	O	B-cell_line
)	NN	O	O
treated	NN	O	O
with	NN	O	O
TNF	NN	O	B-protein
rapidly	NN	O	O
activated	NN	O	O
(	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
)	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
;	NN	O	O
this	NN	O	O
effect	NN	O	O
was	NN	O	O
completely	NN	O	O
abolished	NN	O	O
by	NN	O	O
co-treatment	NN	O	O
with	NN	O	O
phenylarsine	NN	O	O
oxide	NN	O	O
(	NN	O	O
PAO	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
PTPase	NN	O	B-protein
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
VCAM-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
ELAM-1	NN	O	B-protein
by	NN	O	O
TNF	NN	O	B-protein
in	NN	O	O
MVEC	NN	O	B-cell_line
occurred	NN	O	O
within	NN	O	O
6	NN	O	O
h	NN	O	O
and	NN	O	O
was	NN	O	O
also	NN	O	O
completely	NN	O	O
down-regulated	NN	O	O
by	NN	O	O
PAO	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

PAO	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
effective	NN	O	O
even	NN	O	O
when	NN	O	O
added	NN	O	O
3	NN	O	O
h	NN	O	O
after	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
rapid	NN	O	O
mode	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
this	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

Besides	NN	O	O
PAO	NN	O	O
,	NN	O	O
other	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
PTPase	NN	O	B-protein
,	NN	O	O
including	NN	O	O
pervanadate	NN	O	O
and	NN	O	O
diamide	NN	O	O
,	NN	O	O
also	NN	O	O
blocked	NN	O	O
TNF	NN	O	B-protein
-dependent	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
three	NN	O	O
adhesion	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
the	NN	O	O
attachment	NN	O	O
of	NN	O	O
monocytes	NN	O	O
to	NN	O	O
MVEC	NN	O	B-cell_line
was	NN	O	O
also	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
PTPase	NN	O	B-protein
inhibitors	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
overall	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
PTPase	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
either	NN	O	O
directly	NN	O	O
or	NN	O	O
indirectly	NN	O	O
in	NN	O	O
the	NN	O	O
pathway	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
endothelial	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
by	NN	O	O
TNF	NN	O	B-protein
.	NN	O	O

Because	NN	O	O
of	NN	O	O
their	NN	O	O
role	NN	O	O
in	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
,	NN	O	O
PTPase	NN	O	B-protein
may	NN	O	O
provide	NN	O	O
a	NN	O	O
novel	NN	O	O
target	NN	O	O
of	NN	O	O
drug	NN	O	O
development	NN	O	O
for	NN	O	O
treatment	NN	O	O
of	NN	O	O
inflammation	NN	O	O
,	NN	O	O
atherogenesis	NN	O	O
,	NN	O	O
and	NN	O	O
tumor	NN	O	O
metastasis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
its	NN	O	O
inhibitor	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha/MAD-3	NN	O	I-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
by	NN	O	O
Mycobacterium	NN	O	O
tuberculosis	NN	O	O
and	NN	O	O
during	NN	O	O
human	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

Blood	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
active	NN	O	O
tuberculosis	NN	O	O
are	NN	O	O
activated	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
stimulated	NN	O	O
release	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
spontaneous	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
.	NN	O	O

Further	NN	O	O
,	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
demonstrate	NN	O	O
an	NN	O	O
augmented	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
a	NN	O	O
productive	NN	O	O
infection	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Mycobacterium	NN	O	O
tuberculosis	NN	O	O
and	NN	O	O
its	NN	O	O
components	NN	O	O
are	NN	O	O
strong	NN	O	O
signals	NN	O	O
to	NN	O	O
activate	NN	O	O
monocytes	NN	O	B-cell_type
to	NN	O	O
production	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
during	NN	O	O
active	NN	O	O
tuberculosis	NN	O	O
and	NN	O	O
by	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
a	NN	O	O
constitutive	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
,	NN	O	O
the	NN	O	O
major	NN	O	O
cytoplasmic	NN	O	B-protein
inhibitor	NN	O	I-protein
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
in	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
PBMC	NN	O	B-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
or	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
nontuberculous	NN	O	O
pulmonary	NN	O	O
conditions	NN	O	O
were	NN	O	O
intact	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
activated	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
tuberculous	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
gene	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
responsive	NN	O	O
to	NN	O	O
activation	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
was	NN	O	O
up-regulated	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
or	NN	O	O
those	NN	O	O
with	NN	O	O
nontuberculous	NN	O	O
lung	NN	O	O
diseases	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
other	NN	O	O
adherence-associated	NN	O	B-DNA
early	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
IL-8	NN	O	B-protein
and	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
was	NN	O	O
not	NN	O	O
up-regulated	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
of	NN	O	O
tuberculous	NN	O	O
patients	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
and	NN	O	O
its	NN	O	O
tuberculin	NN	O	B-protein
,	NN	O	O
purified	NN	O	B-protein
protein	NN	O	I-protein
derivative	NN	O	I-protein
,	NN	O	O
induced	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B-alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
purified	NN	O	O
protein	NN	O	O
derivative	NN	O	O
induced	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Cooperation	NN	O	O
of	NN	O	O
Spi-1/PU.1	NN	O	B-protein
with	NN	O	O
an	NN	O	O
activated	NN	O	B-protein
erythropoietin	NN	O	I-protein
receptor	NN	O	I-protein
inhibits	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
Epo	NN	O	B-protein
-dependent	NN	O	O
differentiation	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
erythroblasts	NN	O	I-cell_type
and	NN	O	O
induces	NN	O	O
their	NN	O	O
Kit	NN	O	O
ligand-dependent	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Spi-1/PU.1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
myeloid-	NN	O	B-protein
and	NN	O	I-protein
B-cell	NN	O	I-protein
specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
which	NN	O	O
is	NN	O	O
also	NN	O	O
involved	NN	O	O
in	NN	O	O
Friend	NN	O	O
virus-induced	NN	O	O
murine	NN	O	O
erythroleukemia	NN	O	O
.	NN	O	O

The	NN	O	O
pre-leukemic	NN	O	O
phase	NN	O	O
of	NN	O	O
Friend	NN	O	O
erythroleukemia	NN	O	O
results	NN	O	O
from	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
erythropoietin	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
EpoR	NN	O	B-protein
)	NN	O	O
by	NN	O	O
the	NN	O	O
spleen	NN	O	B-protein
focus	NN	O	I-protein
forming	NN	O	I-protein
virus	NN	O	I-protein
(	NN	O	I-protein
SFFV	NN	O	I-protein
)	NN	O	I-protein
envelope	NN	O	I-protein
glycoprotein	NN	O	I-protein
,	NN	O	O
followed	NN	O	O
by	NN	O	O
the	NN	O	O
emergence	NN	O	O
of	NN	O	O
leukemic	NN	O	O
clones	NN	O	O
characterized	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Spi-1	NN	O	B-protein
and	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
p53	NN	O	B-DNA
tumor	NN	O	I-DNA
suppressor	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
developed	NN	O	O
a	NN	O	O
heterologous	NN	O	O
system	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
these	NN	O	O
alterations	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
primary	NN	O	O
erythroblast	NN	O	O
transformation	NN	O	O
.	NN	O	O

Avian	NN	O	B-cell_type
erythroblasts	NN	O	I-cell_type
expressing	NN	O	O
the	NN	O	O
activated	NN	O	B-protein
mouse	NN	O	I-protein
EpoR	NN	O	I-protein
(	NN	O	I-protein
R129C	NN	O	I-protein
)	NN	O	I-protein
differentiated	NN	O	O
into	NN	O	O
erythrocytes	NN	O	B-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
hEpo	NN	O	B-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
Spi-1	NN	O	B-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
inhibited	NN	O	O
this	NN	O	O
ability	NN	O	O
to	NN	O	O
differentiate	NN	O	O
and	NN	O	O
rescued	NN	O	O
the	NN	O	O
cells	NN	O	O
from	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
cell	NN	O	O
death	NN	O	O
program	NN	O	O
normally	NN	O	O
induced	NN	O	O
upon	NN	O	O
hEpo	NN	O	B-protein
withdrawal	NN	O	O
.	NN	O	O

Although	NN	O	O
devoid	NN	O	O
of	NN	O	O
any	NN	O	O
effect	NN	O	O
by	NN	O	O
itself	NN	O	O
,	NN	O	O
a	NN	O	O
mutant	NN	O	B-protein
p53	NN	O	I-protein
cooperated	NN	O	O
with	NN	O	O
Spi-1	NN	O	B-protein
and	NN	O	O
EpoR	NN	O	B-protein
(	NN	O	I-protein
R129C	NN	O	I-protein
)	NN	O	I-protein
to	NN	O	O
reinforce	NN	O	O
both	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
erythroblasts	NN	O	O
co-expressing	NN	O	O
Spi-1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
mouse	NN	O	I-protein
EpoR	NN	O	I-protein
showed	NN	O	O
that	NN	O	O
differentiation	NN	O	O
arrest	NN	O	O
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
depended	NN	O	O
on	NN	O	O
specific	NN	O	O
cooperation	NN	O	O
between	NN	O	O
Spi-1	NN	O	B-protein
and	NN	O	O
EpoR	NN	O	B-protein
(	NN	O	I-protein
R129C	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

This	NN	O	O
cooperation	NN	O	O
was	NN	O	O
also	NN	O	O
required	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
sustained	NN	O	O
proliferation	NN	O	O
of	NN	O	O
differentiation-blocked	NN	O	B-cell_line
erythroblasts	NN	O	I-cell_line
in	NN	O	O
response	NN	O	O
to	NN	O	O
ligand	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
receptor	NN	O	I-protein
c-Kit	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
Spi-1/	NN	O	O
PU.1	NN	O	B-protein
requires	NN	O	O
signals	NN	O	O
emanating	NN	O	O
from	NN	O	O
specific	NN	O	O
cytokine	NN	O	O
and	NN	O	O
growth	NN	O	O
factor	NN	O	O
receptors	NN	O	O
to	NN	O	O
affect	NN	O	O
the	NN	O	O
survival	NN	O	O
,	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
control	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
erythroblasts	NN	O	I-cell_type
.	NN	O	O

They	NN	O	O
also	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
Spi-1/PU.1	NN	O	B-protein
in	NN	O	O
the	NN	O	O
late	NN	O	O
phase	NN	O	O
of	NN	O	O
Friend	NN	O	O
leukemia	NN	O	O
requires	NN	O	O
specific	NN	O	O
signaling	NN	O	O
from	NN	O	O
the	NN	O	O
gp55-modified	NN	O	B-protein
EpoR	NN	O	I-protein
generated	NN	O	O
during	NN	O	O
the	NN	O	O
early	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
viral	NN	O	O
oncogenesis	NN	O	O
and	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
viral	NN	O	O
transcription	NN	O	O
.	NN	O	O

Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
ubiquitous	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
consists	NN	O	O
of	NN	O	O
multiple	NN	O	B-protein
polypeptide	NN	O	I-protein
subunits	NN	O	I-protein
,	NN	O	O
and	NN	O	O
is	NN	O	O
subject	NN	O	O
to	NN	O	O
complex	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
involve	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
,	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
ubiquitination	NN	O	O
,	NN	O	O
proteolytic	NN	O	O
degradation	NN	O	O
,	NN	O	O
and	NN	O	O
nucleocytoplasmic	NN	O	O
translocation	NN	O	O
.	NN	O	O

The	NN	O	O
sophisticated	NN	O	O
control	NN	O	O
of	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
is	NN	O	O
not	NN	O	O
surprising	NN	O	O
since	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
a	NN	O	O
wide	NN	O	O
array	NN	O	O
of	NN	O	O
cellular	NN	O	O
responses	NN	O	O
to	NN	O	O
extracellular	NN	O	O
cues	NN	O	O
,	NN	O	O
associated	NN	O	O
with	NN	O	O
growth	NN	O	O
,	NN	O	O
development	NN	O	O
,	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
and	NN	O	O
pathogen	NN	O	O
invasion	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
unexpected	NN	O	O
that	NN	O	O
this	NN	O	O
versatile	NN	O	O
cellular	NN	O	O
homeostatic	NN	O	O
switch	NN	O	O
would	NN	O	O
be	NN	O	O
affected	NN	O	O
by	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
viral	NN	O	O
pathogens	NN	O	O
,	NN	O	O
which	NN	O	O
have	NN	O	O
evolved	NN	O	O
mechanisms	NN	O	O
to	NN	O	O
utilize	NN	O	O
various	NN	O	O
aspects	NN	O	O
of	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
to	NN	O	O
facilitate	NN	O	O
their	NN	O	O
replication	NN	O	O
,	NN	O	O
cell	NN	O	O
survival	NN	O	O
and	NN	O	O
possibly	NN	O	O
evasion	NN	O	O
of	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

This	NN	O	O
review	NN	O	O
will	NN	O	O
cover	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
are	NN	O	O
utilized	NN	O	O
by	NN	O	O
mammalian	NN	O	O
oncogenic	NN	O	O
viruses	NN	O	O
to	NN	O	O
affect	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
replication	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cellular	NN	O	O
and	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
atopic	NN	O	O
diseases	NN	O	O
:	NN	O	O
a	NN	O	O
study	NN	O	O
with	NN	O	O
allergen-specific	NN	O	B-cell_line
human	NN	O	I-cell_line
helper	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Cytokines	NN	O	B-protein
produced	NN	O	O
by	NN	O	O
helper	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
intimately	NN	O	O
involved	NN	O	O
in	NN	O	O
chronic	NN	O	O
allergic	NN	O	O
diseases	NN	O	O
associated	NN	O	O
with	NN	O	O
eosinophilic	NN	O	O
inflammation	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
We	NN	O	O
investigated	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
,	NN	O	O
a	NN	O	O
potent	NN	O	O
growth	NN	O	O
factor	NN	O	O
and	NN	O	O
chemotactic	NN	O	O
factor	NN	O	O
for	NN	O	O
eosinophils	NN	O	B-cell_type
,	NN	O	O
by	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
asthma	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Allergen-specific	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
and	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
were	NN	O	O
established	NN	O	O
from	NN	O	O
the	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
asthma	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
responses	NN	O	O
to	NN	O	O
various	NN	O	O
stimuli	NN	O	O
were	NN	O	O
determined	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
After	NN	O	O
nonspecific	NN	O	O
stimulation	NN	O	O
,	NN	O	O
IL-5	NN	O	B-protein
production	NN	O	O
by	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
both	NN	O	O
atopic	NN	O	O
and	NN	O	O
nonatopic	NN	O	O
subjects	NN	O	O
with	NN	O	O
asthma	NN	O	O
was	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
compared	NN	O	O
with	NN	O	O
that	NN	O	O
by	NN	O	O
cells	NN	O	O
from	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
atopic	NN	O	O
asthma	NN	O	O
patients	NN	O	O
both	NN	O	O
proliferated	NN	O	O
and	NN	O	O
produced	NN	O	O
IL-5	NN	O	B-protein
after	NN	O	O
incubation	NN	O	O
with	NN	O	O
mite	NN	O	O
allergen	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
mite-specific	NN	O	B-cell_type
helper	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
eosinophilic	NN	O	O
inflammation	NN	O	O
of	NN	O	O
atopic	NN	O	O
asthma	NN	O	O
.	NN	O	O

A	NN	O	O
human	NN	O	O
IL-5	NN	O	B-DNA
promoter/enhancer	NN	O	I-DNA
luciferase	NN	O	I-DNA
gene	NN	O	I-DNA
construct	NN	O	I-DNA
transfected	NN	O	O
into	NN	O	O
IL-5-producing	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
was	NN	O	O
clearly	NN	O	O
transcribed	NN	O	O
after	NN	O	O
stimulation	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
515	NN	O	O
base	NN	O	O
pair	NN	O	O
IL-5	NN	O	B-DNA
gene	NN	O	I-DNA
segment	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
was	NN	O	O
sufficient	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
activating	NN	O	O
signals	NN	O	O
in	NN	O	O
human	NN	O	O
helper	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
same	NN	O	O
gene	NN	O	O
segment	NN	O	O
was	NN	O	O
not	NN	O	O
transcribed	NN	O	O
in	NN	O	O
IL-5-nonproducing	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
IL-5	NN	O	B-protein
synthesis	NN	O	O
is	NN	O	O
regulated	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

Experiments	NN	O	O
with	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
confirmed	NN	O	O
these	NN	O	O
findings	NN	O	O
and	NN	O	O
suggested	NN	O	O
that	NN	O	O
a	NN	O	O
unique	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
may	NN	O	O
be	NN	O	O
essential	NN	O	O
for	NN	O	O
human	NN	O	O
IL-5	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Enhanced	NN	O	O
IL-5	NN	O	B-protein
production	NN	O	O
by	NN	O	O
helper	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
seems	NN	O	O
to	NN	O	O
cause	NN	O	O
the	NN	O	O
eosinophilic	NN	O	O
inflammation	NN	O	O
of	NN	O	O
both	NN	O	O
atopic	NN	O	O
and	NN	O	O
nonatopic	NN	O	O
asthma	NN	O	O
.	NN	O	O

Elucidation	NN	O	O
of	NN	O	O
IL-5	NN	O	B-protein
-specific	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
may	NN	O	O
facilitate	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
novel	NN	O	O
treatments	NN	O	O
for	NN	O	O
allergic	NN	O	O
diseases	NN	O	O
associated	NN	O	O
with	NN	O	O
eosinophilic	NN	O	O
inflammation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cyclosporin	NN	O	O
A	NN	O	O
inhibits	NN	O	O
monocyte	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
cardiac	NN	O	O
transplant	NN	O	O
recipients	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Fibrin	NN	O	O
deposition	NN	O	O
and	NN	O	O
thrombosis	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
both	NN	O	O
allograft	NN	O	O
rejection	NN	O	O
and	NN	O	O
vasculopathy	NN	O	O
after	NN	O	O
cardiac	NN	O	O
transplantation	NN	O	O
.	NN	O	O

Because	NN	O	O
monocytes	NN	O	B-cell_type
play	NN	O	O
a	NN	O	O
pivotal	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
intravascular	NN	O	O
coagulation	NN	O	O
activation	NN	O	O
through	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
synthesize	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
TF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
asked	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
whether	NN	O	O
monocyte	NN	O	O
TF	NN	O	B-protein
activation	NN	O	O
occurs	NN	O	O
in	NN	O	O
cardiac	NN	O	O
transplant	NN	O	O
recipients	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
whether	NN	O	O
monocyte	NN	O	O
TF	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
affected	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
.	NN	O	O

METHODS	NN	O	O
AND	NN	O	O
RESULTS	NN	O	O
:	NN	O	O
We	NN	O	O
measured	NN	O	O
levels	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
highly	NN	O	B-cell_type
purified	NN	O	I-cell_type
monocytes/macrophages	NN	O	I-cell_type
from	NN	O	O
10	NN	O	O
consecutive	NN	O	O
cardiac	NN	O	O
transplant	NN	O	O
recipients	NN	O	O
and	NN	O	O
10	NN	O	O
healthy	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

TF	NN	O	B-protein
activity	NN	O	O
generated	NN	O	O
by	NN	O	O
both	NN	O	O
unstimulated	NN	O	B-cell_line
and	NN	O	O
endotoxin-stimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
in	NN	O	O
transplant	NN	O	O
recipients	NN	O	O
than	NN	O	O
in	NN	O	O
control	NN	O	O
subjects	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
.05	NN	O	O
)	NN	O	O
.	NN	O	O

Increased	NN	O	O
monocyte	NN	O	O
TF	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
transplant	NN	O	O
recipients	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
adversely	NN	O	O
affected	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
CsA	NN	O	O
:	NN	O	O
TF	NN	O	B-protein
induction	NN	O	O
was	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
by	NN	O	O
CsA	NN	O	O
serum	NN	O	O
concentrations	NN	O	O
reaching	NN	O	O
peak	NN	O	O
CsA	NN	O	O
drug	NN	O	O
levels	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
high	NN	O	O
CsA	NN	O	O
blood	NN	O	O
concentrations	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
when	NN	O	O
stimulation	NN	O	O
of	NN	O	O
cells	NN	O	O
was	NN	O	O
performed	NN	O	O
with	NN	O	O
interferon-gamma	NN	O	B-protein
or	NN	O	O
interleukin-1beta	NN	O	B-protein
.	NN	O	O

As	NN	O	O
shown	NN	O	O
by	NN	O	O
reverse	NN	O	O
transcription-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
treatment	NN	O	O
with	NN	O	O
CsA	NN	O	O
leads	NN	O	O
to	NN	O	O
decreased	NN	O	O
TF	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
reduced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
TF	NN	O	B-DNA
promotor	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
This	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
TF	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
occurring	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
cardiac	NN	O	O
transplant	NN	O	O
recipients	NN	O	O
,	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
CsA	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
monocyte	NN	O	O
TF	NN	O	B-protein
induction	NN	O	O
by	NN	O	O
CsA	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
its	NN	O	O
successful	NN	O	O
use	NN	O	O
in	NN	O	O
cardiac	NN	O	O
transplant	NN	O	O
medicine	NN	O	O
and	NN	O	O
might	NN	O	O
be	NN	O	O
useful	NN	O	O
in	NN	O	O
managing	NN	O	O
further	NN	O	O
settings	NN	O	O
of	NN	O	O
vascular	NN	O	O
pathology	NN	O	O
also	NN	O	O
known	NN	O	O
to	NN	O	O
involve	NN	O	O
TF	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

IL-2	NN	O	B-protein
and	NN	O	O
IL-7	NN	O	B-protein
induce	NN	O	O
heterodimerization	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
isoforms	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
.	NN	O	O

Despite	NN	O	O
differences	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
induced	NN	O	O
by	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
and	NN	O	O
IL-7	NN	O	B-protein
,	NN	O	O
both	NN	O	O
cytokines	NN	O	B-protein
modulate	NN	O	O
T	NN	O	O
cell	NN	O	O
functions	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
isoforms	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
STAT5A	NN	O	B-protein
and	NN	O	O
STAT5B	NN	O	B-protein
,	NN	O	O
to	NN	O	O
IL-2-	NN	O	O
and	NN	O	O
IL-7-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
.	NN	O	O

Both	NN	O	O
cytokines	NN	O	B-protein
induced	NN	O	O
assembly	NN	O	O
of	NN	O	O
STAT5A	NN	O	B-protein
and	NN	O	O
STAT5B	NN	O	B-protein
containing	NN	O	O
complexes	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
interferon-gamma	NN	O	B-DNA
activation	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
GAS	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
and	NN	O	O
these	NN	O	O
complexes	NN	O	O
rapidly	NN	O	O
translocated	NN	O	O
(	NN	O	O
within	NN	O	O
1	NN	O	O
min	NN	O	O
)	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
IL-2-	NN	O	B-cell_line
or	NN	O	I-cell_line
IL-7-treated	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
kinetics	NN	O	O
of	NN	O	O
this	NN	O	O
translocation	NN	O	O
were	NN	O	O
delayed	NN	O	O
in	NN	O	O
IL-7-treated	NN	O	B-cell_line
as	NN	O	O
compared	NN	O	O
to	NN	O	O
IL-2-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

IL-2	NN	O	B-protein
and	NN	O	O
IL-7	NN	O	B-protein
were	NN	O	O
equivalent	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT5A	NN	O	B-protein
and	NN	O	O
STAT5B	NN	O	B-protein
and	NN	O	O
to	NN	O	O
facilitate	NN	O	O
binding	NN	O	O
of	NN	O	O
these	NN	O	O
STATs	NN	O	B-protein
to	NN	O	O
an	NN	O	O
immobilized	NN	O	B-DNA
GAS	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-7	NN	O	B-protein
induced	NN	O	O
substantial	NN	O	O
amounts	NN	O	O
of	NN	O	O
STAT5A	NN	O	B-protein
/STAT5B	NN	O	B-protein
heterodimerization	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
constitutive	NN	O	O
association	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
with	NN	O	O
each	NN	O	O
STAT5	NN	O	B-protein
isomer	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-7	NN	O	B-protein
induce	NN	O	O
assembly	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
heterodimers	NN	O	I-protein
in	NN	O	O
a	NN	O	O
similar	NN	O	O
manner	NN	O	O
and	NN	O	O
that	NN	O	O
subsequent	NN	O	O
cellular	NN	O	O
responses	NN	O	O
may	NN	O	O
be	NN	O	O
driven	NN	O	O
by	NN	O	O
induction	NN	O	O
of	NN	O	O
similar	NN	O	O
sets	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
regulates	NN	O	O
inducible	NN	O	O
Oct-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
precursor	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
POU	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
regulate	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
octamer-dependent	NN	O	B-DNA
promoters	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
those	NN	O	O
that	NN	O	O
direct	NN	O	O
transcription	NN	O	O
from	NN	O	O
rearranged	NN	O	O
immunoglobulin	NN	O	B-protein
genes	NN	O	I-protein
.	NN	O	O

Unlike	NN	O	O
Oct-1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
many	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
Oct-2	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
restricted	NN	O	O
primarily	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
precursor	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
precise	NN	O	O
factors	NN	O	O
that	NN	O	O
mediate	NN	O	O
this	NN	O	O
induction	NN	O	O
mechanism	NN	O	O
remain	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
monitored	NN	O	O
Oct-2	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
cells	NN	O	O
arrested	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
an	NN	O	O
LPS-responsive	NN	O	B-protein
member	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Rel	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Despite	NN	O	O
stimulation	NN	O	O
with	NN	O	O
LPS	NN	O	O
,	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
in	NN	O	O
precursor	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
led	NN	O	O
to	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
inducible	NN	O	O
Oct-2	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
Oct-2	NN	O	B-protein
-directed	NN	O	O
transcription	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

This	NN	O	O
biochemical	NN	O	O
defect	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
specific	NN	O	O
block	NN	O	O
to	NN	O	O
Oct-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Oct-1	NN	O	B-protein
was	NN	O	O
unaffected	NN	O	O
.	NN	O	O

The	NN	O	O
finding	NN	O	O
that	NN	O	O
Oct-2	NN	O	B-protein
is	NN	O	O
under	NN	O	O
NF-kappaB	NN	O	B-protein
control	NN	O	O
highlights	NN	O	O
an	NN	O	O
important	NN	O	O
cross-talk	NN	O	O
mechanism	NN	O	O
involving	NN	O	O
two	NN	O	O
distinct	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
families	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
B	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
human	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
trans-activator	NN	O	I-DNA
constructs	NN	O	I-DNA
lacking	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

The	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
trans-activator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
bi-	NN	O	B-protein
or	NN	O	I-protein
multi-functional	NN	O	I-protein
domain	NN	O	I-protein
protein	NN	O	I-protein
which	NN	O	O
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
removal	NN	O	O
of	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
151	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
,	NN	O	O
encompassing	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
acidic	NN	O	B-protein
domain	NN	O	I-protein
but	NN	O	O
leaving	NN	O	O
intact	NN	O	O
the	NN	O	O
proline/serine/threonine-rich	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
mutant	NN	O	O
protein	NN	O	O
with	NN	O	O
potent	NN	O	O
suppressive	NN	O	O
properties	NN	O	O
for	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
stably	NN	O	O
or	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
with	NN	O	O
mutant	NN	O	B-DNA
CIITA	NN	O	I-DNA
constructs	NN	O	I-DNA
showed	NN	O	O
up	NN	O	O
to	NN	O	O
99	NN	O	O
%	NN	O	O
suppression	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigen	NN	O	O
induction	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
marked	NN	O	O
suppression	NN	O	O
of	NN	O	O
HLA-DRA	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
a	NN	O	O
B	NN	O	O
lymphoma	NN	O	O
line	NN	O	O
resulted	NN	O	O
in	NN	O	O
up	NN	O	O
to	NN	O	O
89	NN	O	O
%	NN	O	O
reduction	NN	O	O
of	NN	O	O
constitutive	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
within	NN	O	O
5	NN	O	O
days	NN	O	O
and	NN	O	O
suppression	NN	O	O
of	NN	O	O
HLA-DRA	NN	O	B-RNA
mRNA	NN	O	I-RNA
synthesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Helenalin	NN	O	O
,	NN	O	O
an	NN	O	O
anti-inflammatory	NN	O	O
sesquiterpene	NN	O	O
lactone	NN	O	O
from	NN	O	O
Arnica	NN	O	O
,	NN	O	O
selectively	NN	O	O
inhibits	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Alcoholic	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
form	NN	O	O
Arnicae	NN	O	O
flos	NN	O	O
,	NN	O	O
the	NN	O	O
collective	NN	O	O
name	NN	O	O
for	NN	O	O
flowerheads	NN	O	O
from	NN	O	O
Arnica	NN	O	O
montana	NN	O	O
and	NN	O	O
A.	NN	O	O
chamissonis	NN	O	O
ssp.	NN	O	O
foliosa	NN	O	O
,	NN	O	O
are	NN	O	O
used	NN	O	O
therapeutically	NN	O	O
as	NN	O	O
anti-inflammatory	NN	O	O
remedies	NN	O	O
.	NN	O	O

The	NN	O	O
active	NN	O	O
ingredients	NN	O	O
mediating	NN	O	O
the	NN	O	O
pharmacological	NN	O	O
effect	NN	O	O
are	NN	O	O
mainly	NN	O	O
sesquiterpene	NN	O	O
lactones	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
helenalin	NN	O	O
,	NN	O	O
11alpha	NN	O	O
,	NN	O	O
13-dihydrohelenalin	NN	O	O
,	NN	O	O
chamissonolid	NN	O	O
and	NN	O	O
their	NN	O	O
ester	NN	O	O
derivatives	NN	O	O
.	NN	O	O

While	NN	O	O
these	NN	O	O
compounds	NN	O	O
affect	NN	O	O
various	NN	O	O
cellular	NN	O	O
processes	NN	O	O
,	NN	O	O
current	NN	O	O
data	NN	O	O
do	NN	O	O
not	NN	O	O
fully	NN	O	O
explain	NN	O	O
how	NN	O	O
sesquiterpene	NN	O	O
lactones	NN	O	O
exert	NN	O	O
their	NN	O	O
anti-inflammatory	NN	O	O
effect	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
helenalin	NN	O	O
,	NN	O	O
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
much	NN	O	O
lesser	NN	O	O
degree	NN	O	O
,	NN	O	O
11alpha	NN	O	O
,	NN	O	O
13-dihydrohelenalin	NN	O	O
and	NN	O	O
chamissonolid	NN	O	O
,	NN	O	O
inhibit	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

This	NN	O	O
difference	NN	O	O
in	NN	O	O
efficacy	NN	O	O
,	NN	O	O
which	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
compounds	NN	O	O
'	NN	O	O
anti-inflammatory	NN	O	O
potency	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
differences	NN	O	O
in	NN	O	O
structure	NN	O	O
and	NN	O	O
conformation	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
,	NN	O	O
which	NN	O	O
resides	NN	O	O
in	NN	O	O
an	NN	O	O
inactive	NN	O	O
,	NN	O	O
cytoplasmic	NN	O	B-protein
complex	NN	O	I-protein
in	NN	O	O
unstimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
its	NN	O	O
inhibitory	NN	O	B-protein
subunit	NN	O	I-protein
,	NN	O	O
IkappaB	NN	O	B-protein
.	NN	O	O

Helenalin	NN	O	O
inhibits	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
four	NN	O	O
different	NN	O	O
stimuli	NN	O	O
in	NN	O	O
T-cells	NN	O	B-cell_type
,	NN	O	O
B-cells	NN	O	B-cell_type
and	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
abrogates	NN	O	O
kappaB	NN	O	B-protein
-driven	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
is	NN	O	O
selective	NN	O	O
,	NN	O	O
as	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
four	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
Oct-1	NN	O	B-protein
,	NN	O	O
TBP	NN	O	B-protein
,	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
STAT	NN	O	B-protein
5	NN	O	I-protein
was	NN	O	O
not	NN	O	O
affected	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
inhibition	NN	O	O
is	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
direct	NN	O	O
modification	NN	O	O
of	NN	O	O
the	NN	O	O
active	NN	O	O
NF-kappaB	NN	O	B-protein
heterodimer	NN	O	I-protein
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
helenalin	NN	O	O
modifies	NN	O	O
the	NN	O	O
NF-kappaB/IkappaB	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
preventing	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
IkappaB	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
a	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
anti-inflammatory	NN	O	O
effect	NN	O	O
of	NN	O	O
sesquiterpene	NN	O	O
lactones	NN	O	O
,	NN	O	O
which	NN	O	O
differs	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
other	NN	O	O
nonsteroidal	NN	O	O
anti-inflammatory	NN	O	O
drugs	NN	O	O
(	NN	O	O
NSAIDs	NN	O	O
)	NN	O	O
,	NN	O	O
indomethacin	NN	O	O
and	NN	O	O
acetyl	NN	O	O
salicylic	NN	O	O
acid	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
tax	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
mediates	NN	O	O
the	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
c-sis/platelet-derived	NN	O	B-DNA
growth	NN	O	I-DNA
factor-B	NN	O	I-DNA
promoter	NN	O	I-DNA
through	NN	O	O
interactions	NN	O	O
with	NN	O	O
the	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
Sp1	NN	O	B-protein
and	NN	O	O
NGFI-A/Egr-1	NN	O	B-protein
.	NN	O	O

Transcriptional	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
c-sis/platelet-derived	NN	O	B-DNA
growth	NN	O	I-DNA
factor-B	NN	O	I-DNA
(	NN	O	I-DNA
PDGF-B	NN	O	I-DNA
)	NN	O	I-DNA
proto-oncogene	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
as	NN	O	O
one	NN	O	O
possible	NN	O	O
mechanism	NN	O	O
of	NN	O	O
cellular	NN	O	O
transformation	NN	O	O
by	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
.	NN	O	O

In	NN	O	O
previous	NN	O	O
work	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
an	NN	O	O
essential	NN	O	O
site	NN	O	O
in	NN	O	O
the	NN	O	O
c-sis/PDGF-B	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
Tax-responsive	NN	O	B-DNA
element	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	O
TRE1	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
necessary	NN	O	O
for	NN	O	O
transactivation	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
identified	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
Sp3	NN	O	O
,	NN	O	O
and	NN	O	O
NGFI-A/Egr-1	NN	O	B-protein
as	NN	O	O
the	NN	O	O
primary	NN	O	B-protein
nuclear	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
TRE1	NN	O	B-DNA
which	NN	O	O
mediate	NN	O	O
Tax	NN	O	B-protein
responsiveness	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
work	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
mechanism	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
whereby	NN	O	O
Tax	NN	O	B-protein
transactivates	NN	O	O
the	NN	O	O
c-sis/PDGF-B	NN	O	B-DNA
proto-oncogene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
significantly	NN	O	O
increase	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
template	NN	O	O
containing	NN	O	O
the	NN	O	O
-257	NN	O	B-DNA
to	NN	O	I-DNA
+74	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
c-sis/PDGF-B	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
increased	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
both	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
NGFI-A/Egr-1	NN	O	B-protein
using	NN	O	O
a	NN	O	O
TRE1	NN	O	B-DNA
probe	NN	O	I-DNA
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
mutants	NN	O	I-protein
showed	NN	O	O
that	NN	O	O
two	NN	O	O
mutants	NN	O	O
,	NN	O	O
IEXC29S	NN	O	B-protein
and	NN	O	O
IEXL320G	NN	O	B-protein
,	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
significantly	NN	O	O
transactivate	NN	O	O
the	NN	O	O
c-sis/PDGF-B	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
co-immunoprecipitation	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
stably	NN	O	O
bind	NN	O	O
to	NN	O	O
both	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
NGFI-A/Egr-1	NN	O	B-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
co-immunoprecipitation	NN	O	O
analysis	NN	O	O
also	NN	O	O
revealed	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
mutant	NN	O	O
IEXC29S	NN	O	B-protein
is	NN	O	O
unable	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
NGFI-A/Egr-1	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
Tax	NN	O	B-protein
mutant	NN	O	O
IEXL320G	NN	O	B-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
NGFI-A/Egr-1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
RelB	NN	O	B-protein
is	NN	O	O
associated	NN	O	O
with	NN	O	O
effective	NN	O	O
antigen-presenting	NN	O	O
cell	NN	O	O
function	NN	O	O
.	NN	O	O

Dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
DC	NN	O	B-cell_type
)	NN	O	O
are	NN	O	O
potent	NN	O	O
APCs	NN	O	B-cell_type
that	NN	O	O
enter	NN	O	O
resting	NN	O	O
tissues	NN	O	O
as	NN	O	O
precursors	NN	O	O
and	NN	O	O
,	NN	O	O
after	NN	O	O
Ag	NN	O	O
exposure	NN	O	O
,	NN	O	O
differentiate	NN	O	O
and	NN	O	O
migrate	NN	O	O
to	NN	O	O
draining	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
.	NN	O	O

The	NN	O	O
phenotype	NN	O	O
of	NN	O	O
RelB	NN	O	B-protein
knockout	NN	O	O
mice	NN	O	O
implicates	NN	O	O
this	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B/Rel	NN	O	I-protein
family	NN	O	I-protein
in	NN	O	O
DC	NN	O	B-cell_type
differentiation	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
elucidate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
RelB	NN	O	B-protein
in	NN	O	O
DC	NN	O	B-cell_type
differentiation	NN	O	O
,	NN	O	O
mRNA	NN	O	B-RNA
,	NN	O	O
intracellular	NN	O	O
protein	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
RelB	NN	O	B-protein
were	NN	O	O
examined	NN	O	O
in	NN	O	O
immature	NN	O	O
and	NN	O	O
differentiated	NN	O	O
human	NN	O	B-cell_type
DC	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
other	NN	O	O
PB	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cell	NN	O	I-cell_type
populations	NN	O	I-cell_type
.	NN	O	O

RelB	NN	O	B-protein
protein	NN	O	O
and	NN	O	O
mRNA	NN	O	B-RNA
were	NN	O	O
detected	NN	O	O
constitutively	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
differentiated	NN	O	B-cell_type
DC	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
monocyte-derived	NN	O	B-cell_line
DC	NN	O	I-cell_line
.	NN	O	O

Immunohistochemical	NN	O	O
staining	NN	O	O
demonstrated	NN	O	O
RelB	NN	O	B-protein
within	NN	O	O
the	NN	O	O
differentiated	NN	O	B-cell_type
lymph	NN	O	I-cell_type
node	NN	O	I-cell_type
interdigitating	NN	O	I-cell_type
DC	NN	O	I-cell_type
and	NN	O	O
follicular	NN	O	B-cell_type
DC	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
undifferentiated	NN	O	B-cell_type
DC	NN	O	I-cell_type
in	NN	O	O
normal	NN	O	O
skin	NN	O	O
.	NN	O	O

Active	NN	O	O
nuclear	NN	O	O
RelB	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
by	NN	O	O
supershift	NN	O	O
assay	NN	O	O
only	NN	O	O
in	NN	O	O
differentiated	NN	O	O
DC	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
either	NN	O	O
PB	NN	O	B-cell_type
precursors	NN	O	I-cell_type
or	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
RelB+	NN	O	B-cell_type
APC	NN	O	I-cell_type
were	NN	O	O
potent	NN	O	O
stimulators	NN	O	O
of	NN	O	O
the	NN	O	O
MLR	NN	O	B-protein
.	NN	O	O

The	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
RelB	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
regulated	NN	O	O
both	NN	O	O
transcriptionally	NN	O	O
and	NN	O	O
post-translationally	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Within	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
RelB	NN	O	B-protein
may	NN	O	O
specifically	NN	O	O
transactivate	NN	O	O
genes	NN	O	O
that	NN	O	O
are	NN	O	O
critical	NN	O	O
for	NN	O	O
APC	NN	O	B-cell_type
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-AT	NN	O	B-protein
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
a	NN	O	O
CAML	NN	O	B-protein
-interacting	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
key	NN	O	O
event	NN	O	O
underlying	NN	O	O
lymphocyte	NN	O	O
action	NN	O	O
.	NN	O	O

The	NN	O	O
CAML	NN	O	B-protein
(	NN	O	I-protein
calcium-modulator	NN	O	I-protein
and	NN	O	I-protein
cyclophilin	NN	O	I-protein
ligand	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
a	NN	O	O
coinducer	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
activation	NN	O	O
when	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
was	NN	O	O
isolated	NN	O	O
by	NN	O	O
virtue	NN	O	O
of	NN	O	O
its	NN	O	O
affinity	NN	O	O
for	NN	O	O
CAML	NN	O	B-protein
.	NN	O	O

Cross-linking	NN	O	O
of	NN	O	O
this	NN	O	O
lymphocyte-specific	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
designated	NN	O	O
TACI	NN	O	B-protein
(	NN	O	O
transmembrane	NN	O	B-protein
activator	NN	O	I-protein
and	NN	O	I-protein
CAML-interactor	NN	O	I-protein
)	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
transfected	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
TACI-specific	NN	O	B-protein
antibodies	NN	O	I-protein
led	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
NF-AT	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NFkappaB	NN	O	B-protein
.	NN	O	O

TACI	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
was	NN	O	O
specifically	NN	O	O
blocked	NN	O	O
by	NN	O	O
a	NN	O	O
dominant-negative	NN	O	B-protein
CAML	NN	O	I-protein
mutant	NN	O	I-protein
,	NN	O	O
thus	NN	O	O
implicating	NN	O	O
CAML	NN	O	B-protein
as	NN	O	O
a	NN	O	O
signaling	NN	O	O
intermediate	NN	O	O
.	NN	O	O

-DOCSTART-	O

Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
process	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
used	NN	O	O
to	NN	O	O
eliminate	NN	O	O
unwanted	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
organisms	NN	O	O
.	NN	O	O

Various	NN	O	O
extracellular	NN	O	O
signals	NN	O	O
,	NN	O	O
often	NN	O	O
converging	NN	O	O
in	NN	O	O
common	NN	O	O
intracellular	NN	O	O
pathways	NN	O	O
,	NN	O	O
can	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
a	NN	O	O
cell-type-specific	NN	O	O
fashion	NN	O	O
.	NN	O	O

Recent	NN	O	O
work	NN	O	O
from	NN	O	O
several	NN	O	O
laboratories	NN	O	O
has	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
regulate	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
many	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

In	NN	O	O
most	NN	O	O
cells	NN	O	O
,	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
appear	NN	O	O
to	NN	O	O
mediate	NN	O	O
survival	NN	O	O
signals	NN	O	O
that	NN	O	O
protect	NN	O	O
cells	NN	O	O
from	NN	O	O
apoptosis	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
under	NN	O	O
some	NN	O	O
circumstances	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
may	NN	O	O
also	NN	O	O
promote	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
thiol	NN	O	O
antioxidant	NN	O	O
regulates	NN	O	O
IgE	NN	O	B-protein
isotype	NN	O	I-protein
switching	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
.	NN	O	O

The	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
is	NN	O	O
present	NN	O	O
at	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
germline	NN	O	B-DNA
Cepsilon	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
there	NN	O	O
is	NN	O	O
little	NN	O	O
information	NN	O	O
on	NN	O	O
whether	NN	O	O
this	NN	O	O
factor	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
regulating	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Accordingly	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
germline	NN	O	B-DNA
Cepsilon	NN	O	O
transcription	NN	O	O
by	NN	O	O
using	NN	O	O
two	NN	O	O
human	NN	O	B-cell_line
Burkitt	NN	O	I-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
DND39	NN	O	B-cell_line
and	NN	O	O
DG75	NN	O	B-cell_line
.	NN	O	O

In	NN	O	O
both	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
n-acetyl-L-cysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
thiol	NN	O	O
antioxidant	NN	O	O
,	NN	O	O
inhibited	NN	O	O
the	NN	O	O
triggering	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
by	NN	O	O
anti-CD40	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
IL-4	NN	O	B-protein
activated	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
6	NN	O	I-protein
in	NN	O	O
addition	NN	O	O
to	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
NAC	NN	O	O
treatment	NN	O	O
or	NN	O	O
the	NN	O	O
transfection	NN	O	O
of	NN	O	O
decoy	NN	O	O
oligodeoxynucleotides	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
or	NN	O	O
STAT6	NN	O	B-protein
only	NN	O	O
partly	NN	O	O
blocked	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
germline	NN	O	B-DNA
Cepsilon	NN	O	O
transcription	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
these	NN	O	O
two	NN	O	O
decoy	NN	O	O
oligodeoxynucleotides	NN	O	O
together	NN	O	O
almost	NN	O	O
completely	NN	O	O
abrogated	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
germline	NN	O	B-DNA
Cepsilon	NN	O	O
transcription	NN	O	O
.	NN	O	O

Of	NN	O	O
note	NN	O	O
,	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
enhancement	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
-driven	NN	O	O
germline	NN	O	B-DNA
Cepsilon	NN	O	O
transcription	NN	O	O
was	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
by	NN	O	O
NAC	NN	O	O
or	NN	O	O
by	NN	O	O
a	NN	O	O
decoy	NN	O	O
oligodeoxynucleotide	NN	O	O
for	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
NAC	NN	O	O
was	NN	O	O
also	NN	O	O
examined	NN	O	O
on	NN	O	O
deletional	NN	O	O
switch	NN	O	O
recombination	NN	O	O
underlying	NN	O	O
the	NN	O	O
isotype	NN	O	O
switch	NN	O	O
to	NN	O	O
IgE	NN	O	B-protein
.	NN	O	O

NAC	NN	O	O
inhibited	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
Smu/Sepsilon	NN	O	B-protein
switch	NN	O	I-protein
fragments	NN	O	I-protein
in	NN	O	O
normal	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
costimulated	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
anti-CD40	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
.	NN	O	O

It	NN	O	O
also	NN	O	O
abolished	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
upregulation	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
but	NN	O	O
promoted	NN	O	O
upregulation	NN	O	O
of	NN	O	O
CD23	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
coordination	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
STAT6	NN	O	B-protein
may	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
germline	NN	O	B-DNA
Cepsilon	NN	O	O
transcription	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
regulating	NN	O	O
both	NN	O	O
enhancement	NN	O	O
of	NN	O	O
germline	NN	O	B-DNA
Cepsilon	NN	O	O
transcription	NN	O	O
and	NN	O	O
class	NN	O	O
switching	NN	O	O
to	NN	O	O
IgE	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Pathogenesis	NN	O	O
.	NN	O	O

There	NN	O	O
are	NN	O	O
many	NN	O	O
hypotheses	NN	O	O
concerning	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
endometriosis	NN	O	O
,	NN	O	O
though	NN	O	O
no	NN	O	O
single	NN	O	O
theory	NN	O	O
can	NN	O	O
explain	NN	O	O
all	NN	O	O
cases	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
likely	NN	O	O
that	NN	O	O
several	NN	O	O
mechanisms	NN	O	O
are	NN	O	O
involved	NN	O	O
.	NN	O	O

Early	NN	O	O
studies	NN	O	O
concentrated	NN	O	O
on	NN	O	O
the	NN	O	O
histogenesis	NN	O	O
of	NN	O	O
the	NN	O	O
endometriotic	NN	O	O
lesion	NN	O	O
.	NN	O	O

Recent	NN	O	O
evidence	NN	O	O
has	NN	O	O
implicated	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
endometriosis	NN	O	O
.	NN	O	O

This	NN	O	O
review	NN	O	O
considers	NN	O	O
the	NN	O	O
evidence	NN	O	O
for	NN	O	O
different	NN	O	O
theories	NN	O	O
of	NN	O	O
the	NN	O	O
histogenesis	NN	O	O
of	NN	O	O
endometriosis	NN	O	O
and	NN	O	O
discusses	NN	O	O
possible	NN	O	O
immune	NN	O	O
factors	NN	O	O
that	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

ATF1	NN	O	B-protein
and	NN	O	O
CREB	NN	O	B-protein
trans-activate	NN	O	O
a	NN	O	O
cell	NN	O	B-DNA
cycle	NN	O	I-DNA
regulated	NN	O	I-DNA
histone	NN	O	I-DNA
H4	NN	O	I-DNA
gene	NN	O	I-DNA
at	NN	O	O
a	NN	O	O
distal	NN	O	B-DNA
nuclear	NN	O	I-DNA
matrix	NN	O	I-DNA
associated	NN	O	I-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Proteins	NN	O	O
of	NN	O	O
the	NN	O	O
ATF/	NN	O	O
CREB	NN	O	B-protein
class	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
stimulate	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
cell	NN	O	B-DNA
growth-related	NN	O	I-DNA
genes	NN	O	I-DNA
through	NN	O	O
protein	NN	O	B-DNA
kinase	NN	O	I-DNA
A-related	NN	O	I-DNA
cAMP	NN	O	I-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
promoter	NN	O	O
activity	NN	O	O
of	NN	O	O
cell	NN	O	B-DNA
cycle	NN	O	I-DNA
regulated	NN	O	I-DNA
histone	NN	O	I-DNA
H4	NN	O	I-DNA
genes	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
at	NN	O	O
least	NN	O	O
four	NN	O	O
principal	NN	O	B-DNA
cis-acting	NN	O	I-DNA
elements	NN	O	I-DNA
which	NN	O	O
mediate	NN	O	O
G1/S	NN	O	O
phase	NN	O	O
control	NN	O	O
and/or	NN	O	O
enhancement	NN	O	O
of	NN	O	O
transcription	NN	O	O
during	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

Using	NN	O	O
protein-DNA	NN	O	O
interaction	NN	O	O
assays	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
H4	NN	O	B-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
two	NN	O	O
ATF/	NN	O	O
CREB	NN	O	B-protein
recognition	NN	O	O
motifs	NN	O	O
which	NN	O	O
interact	NN	O	O
with	NN	O	O
CREB	NN	O	B-protein
,	NN	O	O
ATF1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
ATF2	NN	O	B-protein
but	NN	O	O
not	NN	O	O
with	NN	O	O
ATF4/CREB2	NN	O	B-protein
.	NN	O	O

One	NN	O	O
ATF/CRE	NN	O	B-DNA
motif	NN	O	I-DNA
is	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
promoter	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
nuclear	NN	O	B-DNA
matrix-associated	NN	O	I-DNA
Site	NN	O	I-DNA
IV	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
second	NN	O	O
motif	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
at	NN	O	O
Site	NN	O	B-DNA
I	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
ATF/CRE	NN	O	B-DNA
motifs	NN	O	I-DNA
overlap	NN	O	O
binding	NN	O	B-DNA
sequences	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
multifunctional	NN	O	B-protein
YY1	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
previously	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
associated	NN	O	O
.	NN	O	O

Subnuclear	NN	O	O
fractionation	NN	O	O
reveals	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
two	NN	O	O
ATF1	NN	O	B-protein
isoforms	NN	O	I-protein
which	NN	O	O
appear	NN	O	O
to	NN	O	O
differ	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
partition	NN	O	O
selectively	NN	O	O
between	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
and	NN	O	O
nonmatrix	NN	O	O
compartments	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
in	NN	O	O
regulating	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Site-directed	NN	O	O
mutational	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Site	NN	O	B-DNA
I	NN	O	I-DNA
and	NN	O	O
Site	NN	O	B-DNA
IV	NN	O	I-DNA
together	NN	O	O
support	NN	O	O
ATF1-	NN	O	O
and	NN	O	O
CREB-induced	NN	O	O
trans-activation	NN	O	O
of	NN	O	O
the	NN	O	O
H4	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
establish	NN	O	O
that	NN	O	O
ATF/CREB	NN	O	B-protein
factors	NN	O	I-protein
functionally	NN	O	O
modulate	NN	O	O
histone	NN	O	B-DNA
H4	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
at	NN	O	O
distal	NN	O	B-DNA
and	NN	O	I-DNA
proximal	NN	O	I-DNA
promoter	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Is	NN	O	O
celiac	NN	O	O
disease	NN	O	O
due	NN	O	O
to	NN	O	O
molecular	NN	O	O
mimicry	NN	O	O
between	NN	O	O
gliadin	NN	O	O
peptide	NN	O	O
-HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecule	NN	O	I-protein
-T	NN	O	O
cell	NN	O	O
interactions	NN	O	O
and	NN	O	O
those	NN	O	O
of	NN	O	O
some	NN	O	O
unidentified	NN	O	B-protein
superantigen	NN	O	I-protein
?	NN	O	O

This	NN	O	O
paper	NN	O	O
presents	NN	O	O
a	NN	O	O
new	NN	O	O
hypothesis	NN	O	O
for	NN	O	O
the	NN	O	O
etiology	NN	O	O
and	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
celiac	NN	O	O
disease	NN	O	O
(	NN	O	O
CD	NN	O	O
)	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
our	NN	O	O
contention	NN	O	O
that	NN	O	O
CD	NN	O	O
is	NN	O	O
triggered	NN	O	O
by	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
gliadin	NN	O	O
peptides	NN	O	O
to	NN	O	O
CD-associated	NN	O	B-protein
HLA	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
these	NN	O	O
putative	NN	O	O
CD	NN	O	O
peptides	NN	O	O
bind	NN	O	O
to	NN	O	O
oligosaccharide	NN	O	O
residues	NN	O	O
on	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
distal	NN	O	O
to	NN	O	O
the	NN	O	O
peptide-binding	NN	O	B-protein
groove	NN	O	I-protein
invoking	NN	O	O
recognition	NN	O	O
and	NN	O	O
binding	NN	O	O
by	NN	O	O
specialized	NN	O	O
subsets	NN	O	O
of	NN	O	O
gamma	NN	O	B-cell_type
delta	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
receptor-bearing	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
these	NN	O	O
gamma	NN	O	B-cell_type
delta	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
serves	NN	O	O
as	NN	O	O
a	NN	O	O
signal	NN	O	O
for	NN	O	O
abrogation	NN	O	O
of	NN	O	O
oral	NN	O	O
tolerance	NN	O	O
to	NN	O	O
ingested	NN	O	B-protein
proteins	NN	O	I-protein
setting	NN	O	O
in	NN	O	O
motion	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
immune	NN	O	O
responses	NN	O	O
directed	NN	O	O
against	NN	O	O
the	NN	O	O
small	NN	O	O
intestinal	NN	O	O
epithelium	NN	O	O
of	NN	O	O
CD	NN	O	O
patients	NN	O	O
.	NN	O	O

CD	NN	O	O
patients	NN	O	O
are	NN	O	O
victimized	NN	O	O
by	NN	O	O
this	NN	O	O
self-distructed	NN	O	O
immune	NN	O	O
response	NN	O	O
because	NN	O	O
of	NN	O	O
inheritance	NN	O	O
of	NN	O	O
certain	NN	O	O
combinations	NN	O	O
of	NN	O	O
HLA-DQ	NN	O	O
and	NN	O	O
DR	NN	O	O
haplotypes	NN	O	O
.	NN	O	O

Dimers	NN	O	O
encoded	NN	O	O
by	NN	O	O
HLA-DR	NN	O	B-protein
haplotypes	NN	O	O
may	NN	O	O
be	NN	O	O
the	NN	O	O
primary	NN	O	B-DNA
restriction	NN	O	I-DNA
elements	NN	O	I-DNA
for	NN	O	O
lectin-like	NN	O	O
,	NN	O	O
gliadin	NN	O	O
peptides	NN	O	O
while	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
immune	NN	O	O
suppression	NN	O	O
(	NN	O	O
or	NN	O	O
lack	NN	O	O
thereof	NN	O	O
)	NN	O	O
to	NN	O	O
ingested	NN	O	O
gliadins	NN	O	O
is	NN	O	O
governed	NN	O	O
by	NN	O	O
inherited	NN	O	O
HLA-DQ	NN	O	B-DNA
haplotypes	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
speculate	NN	O	O
that	NN	O	O
molecular	NN	O	O
mimicry	NN	O	O
between	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
gliadin	NN	O	O
peptides	NN	O	O
and	NN	O	O
some	NN	O	O
,	NN	O	O
as	NN	O	O
yet	NN	O	O
unidentified	NN	O	O
,	NN	O	O
bacterial	NN	O	B-protein
or	NN	O	I-protein
viral	NN	O	I-protein
superantigen	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
disease	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	O
virus	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein-1	NN	O	I-protein
triggers	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
via	NN	O	O
the	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	O
kinase	NN	O	O
cascade	NN	O	O
.	NN	O	O

The	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
LMP-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
integral	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
which	NN	O	O
transforms	NN	O	O
fibroblasts	NN	O	B-cell_type
and	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
EBV-mediated	NN	O	O
B-cell	NN	O	O
immortalization	NN	O	O
.	NN	O	O

LMP-1	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
trigger	NN	O	O
cellular	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
which	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
can	NN	O	O
not	NN	O	O
fully	NN	O	O
explain	NN	O	O
the	NN	O	O
oncogenic	NN	O	O
potential	NN	O	O
of	NN	O	O
LMP-1	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
LMP-1	NN	O	B-protein
induces	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
a	NN	O	O
dimer	NN	O	O
of	NN	O	O
Jun/Jun	NN	O	B-protein
or	NN	O	I-protein
Jun/Fos	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

LMP-1	NN	O	B-protein
effects	NN	O	O
on	NN	O	O
AP-1	NN	O	B-protein
are	NN	O	O
mediated	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	O
kinase	NN	O	O
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
cascade	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
Erk	NN	O	B-protein
)	NN	O	O
pathway	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
LMP-1	NN	O	B-protein
triggers	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
transactivation	NN	O	I-protein
domain	NN	O	I-protein
which	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
activated	NN	O	O
upon	NN	O	O
JNK	NN	O	B-protein
-mediated	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

Deletion	NN	O	O
analysis	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
55	NN	O	O
C-terminal	NN	O	B-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
of	NN	O	O
the	NN	O	O
LMP-1	NN	O	B-protein
molecule	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
its	NN	O	O
TRAF	NN	O	B-protein
interaction	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
are	NN	O	O
essential	NN	O	O
for	NN	O	O
AP-1	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

JNK	NN	O	B-protein
-mediated	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
the	NN	O	O
direct	NN	O	O
output	NN	O	O
of	NN	O	O
LMP	NN	O	B-protein
-1-triggered	NN	O	O
signaling	NN	O	O
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
an	NN	O	O
inducible	NN	O	O
LMP-1	NN	O	B-protein
mutant	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
a	NN	O	O
tetracycline-regulated	NN	O	B-DNA
LMP-1	NN	O	I-DNA
allele	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
JNK	NN	O	B-protein
is	NN	O	O
also	NN	O	O
an	NN	O	O
effector	NN	O	O
of	NN	O	O
non-cytotoxic	NN	O	O
LMP-1	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
physiological	NN	O	O
target	NN	O	O
cells	NN	O	O
of	NN	O	O
EBV	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
reveal	NN	O	O
a	NN	O	O
novel	NN	O	O
effector	NN	O	O
of	NN	O	O
LMP-1	NN	O	B-protein
,	NN	O	O
the	NN	O	O
SEK/	NN	O	O
JNK	NN	O	B-protein
/	NN	O	O
c-Jun	NN	O	B-protein
/	NN	O	O
AP-1	NN	O	B-protein
pathway	NN	O	O
,	NN	O	O
which	NN	O	O
contributes	NN	O	O
to	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
immortalizing	NN	O	O
and	NN	O	O
transforming	NN	O	O
potential	NN	O	O
of	NN	O	O
LMP-1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Abnormal	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
in	NN	O	O
humans	NN	O	O
exposed	NN	O	O
to	NN	O	O
methyl	NN	O	O
tertiary-butyl	NN	O	O
ether	NN	O	O
and	NN	O	O
benzene	NN	O	O
contaminating	NN	O	O
water	NN	O	O
.	NN	O	O

1.	NN	O	O
In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
in	NN	O	O
individuals	NN	O	O
with	NN	O	O
certain	NN	O	O
genetic	NN	O	O
makeup	NN	O	O
,	NN	O	O
MTBE	NN	O	O
,	NN	O	O
benzene	NN	O	O
or	NN	O	O
their	NN	O	O
metabolites	NN	O	O
act	NN	O	O
as	NN	O	O
adducts	NN	O	O
and	NN	O	O
may	NN	O	O
induce	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

2.	NN	O	O
Our	NN	O	O
study	NN	O	O
involved	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
60	NN	O	O
male	NN	O	O
and	NN	O	O
female	NN	O	O
subjects	NN	O	O
who	NN	O	O
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
MTBE	NN	O	O
and	NN	O	O
benzene-contaminated	NN	O	O
water	NN	O	O
concentrations	NN	O	O
up	NN	O	O
to	NN	O	O
76	NN	O	O
PPB	NN	O	O
for	NN	O	O
MTBE	NN	O	O
and	NN	O	O
14	NN	O	O
PPB	NN	O	O
for	NN	O	O
benzene	NN	O	O
,	NN	O	O
for	NN	O	O
a	NN	O	O
period	NN	O	O
of	NN	O	O
5	NN	O	O
to	NN	O	O
8	NN	O	O
years	NN	O	O
.	NN	O	O

For	NN	O	O
comparison	NN	O	O
,	NN	O	O
we	NN	O	O
recruited	NN	O	O
a	NN	O	O
control	NN	O	O
group	NN	O	O
consisting	NN	O	O
of	NN	O	O
32	NN	O	O
healthy	NN	O	O
males	NN	O	O
and	NN	O	O
females	NN	O	O
with	NN	O	O
similar	NN	O	O
age	NN	O	O
distribution	NN	O	O
and	NN	O	O
without	NN	O	O
a	NN	O	O
history	NN	O	O
of	NN	O	O
exposure	NN	O	O
to	NN	O	O
MTBE	NN	O	O
or	NN	O	O
benzene	NN	O	O
.	NN	O	O

3	NN	O	O
.	NN	O	O
Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
PBL	NN	O	B-cell_type
)	NN	O	O
of	NN	O	O
both	NN	O	O
groups	NN	O	O
were	NN	O	O
tested	NN	O	O
for	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
using	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

4.	NN	O	O
When	NN	O	O
apoptotic	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
exposed	NN	O	O
individuals	NN	O	O
were	NN	O	O
compared	NN	O	O
to	NN	O	O
apoptotic	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
control	NN	O	O
group	NN	O	O
,	NN	O	O
statistically-significant	NN	O	O
differences	NN	O	O
between	NN	O	O
each	NN	O	O
mean	NN	O	O
group	NN	O	O
were	NN	O	O
detected	NN	O	O
(	NN	O	O
26.4	NN	O	O
+/-	NN	O	O
1.8	NN	O	O
and	NN	O	O
12.1	NN	O	O
+/-	NN	O	O
1.3	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
,	NN	O	O
indicating	NN	O	O
an	NN	O	O
increased	NN	O	O
rate	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
80.5	NN	O	O
%	NN	O	O
of	NN	O	O
exposed	NN	O	O
individuals	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.0001	NN	O	O
,	NN	O	O
Mann-Whitney	NN	O	O
U-Test	NN	O	O
)	NN	O	O
.	NN	O	O

MTBE	NN	O	O
and	NN	O	O
benzene-induced	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
attributed	NN	O	O
to	NN	O	O
a	NN	O	O
discrete	NN	O	O
block	NN	O	O
within	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
.	NN	O	O

Because	NN	O	O
cell	NN	O	O
cycle	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
in	NN	O	O
PBL	NN	O	B-cell_type
from	NN	O	O
chemically-exposed	NN	O	O
individuals	NN	O	O
,	NN	O	O
between	NN	O	O
20-50	NN	O	O
%	NN	O	O
of	NN	O	O
cells	NN	O	O
were	NN	O	O
accumulated	NN	O	O
at	NN	O	O
the	NN	O	O
S-G2/M	NN	O	O
boundaries	NN	O	O
.	NN	O	O

5.	NN	O	O
One	NN	O	O
of	NN	O	O
the	NN	O	O
signaling	NN	O	B-protein
molecules	NN	O	I-protein
which	NN	O	O
mediates	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
is	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
Kappa-B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
examined	NN	O	O
as	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
many	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
mediating	NN	O	O
cell	NN	O	O
death	NN	O	O
by	NN	O	O
MTBE	NN	O	O
and	NN	O	O
benzene	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
,	NN	O	O
to	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
chemically-exposed	NN	O	O
group	NN	O	O
was	NN	O	O
capable	NN	O	O
of	NN	O	O
inhibiting	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
by	NN	O	O
40	NN	O	O
%	NN	O	O
.	NN	O	O

This	NN	O	O
reversal	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
almost	NN	O	O
to	NN	O	O
the	NN	O	O
control	NN	O	O
level	NN	O	O
by	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
may	NN	O	O
indicate	NN	O	O
involvement	NN	O	O
of	NN	O	O
this	NN	O	O
signaling	NN	O	B-protein
molecule	NN	O	I-protein
in	NN	O	O
MTBE	NN	O	O
and	NN	O	O
benzene	NN	O	O
induction	NN	O	O
of	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

-DOCSTART-	O

Purification	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
SR	NN	O	I-protein
31747A-binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

A	NN	O	O
nuclear	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
related	NN	O	O
to	NN	O	O
yeast	NN	O	B-protein
sterol	NN	O	I-protein
isomerase	NN	O	I-protein
.	NN	O	O

SR	NN	O	O
31747A	NN	O	O
,	NN	O	O
defined	NN	O	O
as	NN	O	O
a	NN	O	O
sigma	NN	O	O
ligand	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
novel	NN	O	O
immunosuppressive	NN	O	O
agent	NN	O	O
that	NN	O	O
blocks	NN	O	O
proliferation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
and	NN	O	I-cell_type
mouse	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
a	NN	O	O
radiolabeled	NN	O	O
chemical	NN	O	O
probe	NN	O	O
,	NN	O	O
we	NN	O	O
here	NN	O	O
purified	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
SR	NN	O	O
31747A	NN	O	O
and	NN	O	O
called	NN	O	O
it	NN	O	O
SR	NN	O	B-protein
31747A-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
SR-BP	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Purified	NN	O	B-protein
SR-BP	NN	O	I-protein
retained	NN	O	O
its	NN	O	O
binding	NN	O	O
properties	NN	O	O
and	NN	O	O
migrated	NN	O	O
on	NN	O	O
SDS-polyacrylamide	NN	O	O
gel	NN	O	O
as	NN	O	O
a	NN	O	O
Mr	NN	O	B-protein
28	NN	O	I-protein
,	NN	O	I-protein
000	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Cloning	NN	O	O
of	NN	O	O
the	NN	O	O
cDNA	NN	O	B-DNA
encoding	NN	O	O
human	NN	O	B-protein
SR-BP	NN	O	I-protein
shows	NN	O	O
an	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
for	NN	O	O
a	NN	O	O
223-amino	NN	O	B-protein
acid	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
recently	NN	O	O
cloned	NN	O	O
sigma	NN	O	B-protein
1	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
deduced	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
fungal	NN	O	B-protein
C8-C7	NN	O	I-protein
sterol	NN	O	I-protein
isomerase	NN	O	I-protein
,	NN	O	O
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
ERG2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
ERG2	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
has	NN	O	O
been	NN	O	O
identified	NN	O	O
recently	NN	O	O
as	NN	O	O
the	NN	O	O
molecular	NN	O	O
target	NN	O	O
of	NN	O	O
SR	NN	O	O
31747A	NN	O	O
that	NN	O	O
mediates	NN	O	O
antiproliferative	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
drug	NN	O	O
in	NN	O	O
yeast	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
SR-BP	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
revealed	NN	O	O
a	NN	O	O
single	NN	O	O
transcript	NN	O	O
of	NN	O	O
2	NN	O	B-RNA
kilobases	NN	O	I-RNA
which	NN	O	O
was	NN	O	O
widely	NN	O	O
expressed	NN	O	O
among	NN	O	O
organs	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
highest	NN	O	O
abundance	NN	O	O
in	NN	O	O
liver	NN	O	O
and	NN	O	O
the	NN	O	O
lowest	NN	O	O
abundance	NN	O	O
in	NN	O	O
brain	NN	O	O
.	NN	O	O

Subcellular	NN	O	O
localization	NN	O	O
analysis	NN	O	O
in	NN	O	O
various	NN	O	O
cells	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
specific	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
raised	NN	O	O
against	NN	O	O
SR-BP	NN	O	B-protein
,	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
this	NN	O	O
protein	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
nuclear	NN	O	O
envelope	NN	O	O
.	NN	O	O

When	NN	O	O
studying	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
SR	NN	O	O
31747A	NN	O	O
on	NN	O	O
membranes	NN	O	O
from	NN	O	O
yeast	NN	O	O
expressing	NN	O	O
SR-BP	NN	O	B-protein
,	NN	O	O
we	NN	O	O
found	NN	O	O
a	NN	O	O
pharmacological	NN	O	O
profile	NN	O	O
of	NN	O	O
sigma	NN	O	B-protein
1	NN	O	I-protein
receptors	NN	O	I-protein
;	NN	O	O
binding	NN	O	O
was	NN	O	O
displaced	NN	O	O
by	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-pentazocine	NN	O	O
,	NN	O	O
haloperidol	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-SKF	NN	O	O
10	NN	O	O
,	NN	O	O
047	NN	O	O
,	NN	O	O
with	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-SKF	NN	O	O
10	NN	O	O
,	NN	O	O
047	NN	O	O
being	NN	O	O
a	NN	O	O
more	NN	O	O
potent	NN	O	O
competitor	NN	O	O
than	NN	O	O
(	NN	O	O
-	NN	O	O
)	NN	O	O
-SKF	NN	O	O
10	NN	O	O
,	NN	O	O
047	NN	O	O
.	NN	O	O

Scatchard	NN	O	O
plot	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
Kd	NN	O	O
values	NN	O	O
of	NN	O	O
7.1	NN	O	O
nM	NN	O	O
and	NN	O	O
0.15	NN	O	O
nM	NN	O	O
for	NN	O	O
(	NN	O	O
+	NN	O	O
)	NN	O	O
-pentazocine	NN	O	O
and	NN	O	O
SR	NN	O	O
31747A	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
indicating	NN	O	O
an	NN	O	O
affinity	NN	O	O
of	NN	O	O
SR-BP	NN	O	B-protein
50-fold	NN	O	O
higher	NN	O	O
for	NN	O	O
SR	NN	O	O
31747A	NN	O	O
than	NN	O	O
for	NN	O	O
pentazocine	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
pentazocine	NN	O	O
,	NN	O	O
a	NN	O	O
competitive	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
SR	NN	O	O
31747A	NN	O	O
binding	NN	O	O
,	NN	O	O
also	NN	O	O
prevents	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
effect	NN	O	O
of	NN	O	O
SR	NN	O	O
31747A	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
SR-BP	NN	O	B-protein
represents	NN	O	O
the	NN	O	O
molecular	NN	O	O
target	NN	O	O
for	NN	O	O
SR	NN	O	O
31747A	NN	O	O
in	NN	O	O
mammalian	NN	O	O
tissues	NN	O	O
,	NN	O	O
which	NN	O	O
could	NN	O	O
be	NN	O	O
critical	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Phospholipase	NN	O	B-protein
C	NN	O	I-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
overexpression	NN	O	O
and	NN	O	O
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
beta	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
an	NN	O	O
ISGF3	NN	O	B-protein
-independent	NN	O	O
response	NN	O	O
.	NN	O	O

Interferons	NN	O	B-protein
exert	NN	O	O
their	NN	O	O
antiviral	NN	O	O
,	NN	O	O
antiproliferative	NN	O	O
and	NN	O	O
immunoregulatory	NN	O	B-protein
activities	NN	O	O
by	NN	O	O
stimulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
genes	NN	O	O
.	NN	O	O

Such	NN	O	O
genes	NN	O	O
disclose	NN	O	O
a	NN	O	O
common	NN	O	O
element	NN	O	O
within	NN	O	O
their	NN	O	O
promoters	NN	O	B-DNA
,	NN	O	O
defined	NN	O	O
Interferon	NN	O	B-DNA
Stimulated	NN	O	I-DNA
Response	NN	O	I-DNA
Element	NN	O	I-DNA
(	NN	O	O
ISRE	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
which	NN	O	O
binds	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
translocated	NN	O	O
from	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
(	NN	O	O
ISGF3	NN	O	B-protein
)	NN	O	O
after	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
(	NN	O	O
IFN	NN	O	B-protein
)	NN	O	O
to	NN	O	O
the	NN	O	O
specific	NN	O	O
receptor	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
synthesis	NN	O	O
and	NN	O	O
of	NN	O	O
the	NN	O	O
hydrolytic	NN	O	O
activity	NN	O	O
of	NN	O	O
phospholipase	NN	O	B-protein
C	NN	O	I-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
PLC	NN	O	B-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
IFN-beta	NN	O	B-protein
.	NN	O	O

The	NN	O	O
increased	NN	O	O
level	NN	O	O
of	NN	O	O
PLC	NN	O	B-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
becomes	NN	O	O
evident	NN	O	O
after	NN	O	O
90	NN	O	O
min	NN	O	O
of	NN	O	O
IFN-beta	NN	O	B-protein
treatment	NN	O	O
and	NN	O	O
is	NN	O	O
still	NN	O	O
detectable	NN	O	O
after	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

Neither	NN	O	O
the	NN	O	O
PLC	NN	O	B-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
overexpression	NN	O	O
induced	NN	O	O
by	NN	O	O
IFN	NN	O	B-protein
nor	NN	O	O
the	NN	O	O
increased	NN	O	O
hydrolytic	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	B-protein
appear	NN	O	O
to	NN	O	O
be	NN	O	O
affected	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
the	NN	O	O
protein	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
inhibitor	NN	O	O
genistein	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
prevent	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
ISGF3	NN	O	B-protein
components	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
IFN-beta	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
distinct	NN	O	O
from	NN	O	O
ISGF3	NN	O	B-protein
to	NN	O	O
regulate	NN	O	O
PLC	NN	O	B-protein
gamma	NN	O	I-protein
1	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
this	NN	O	O
enzyme	NN	O	B-protein
to	NN	O	O
hydrolyse	NN	O	O
PIP2	NN	O	O
,	NN	O	O
also	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
genistein	NN	O	O
,	NN	O	O
implies	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
this	NN	O	O
enzyme	NN	O	O
can	NN	O	O
exert	NN	O	O
its	NN	O	O
hydrolytic	NN	O	O
activity	NN	O	O
independently	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Negative	NN	O	O
regulation	NN	O	O
by	NN	O	O
HLA-DO	NN	O	B-protein
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
-restricted	NN	O	O
antigen	NN	O	O
processing	NN	O	O
.	NN	O	O

HLA-DM	NN	O	B-protein
is	NN	O	O
a	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	I-protein
MHC	NN	O	I-protein
)	NN	O	I-protein
class	NN	O	I-protein
II-like	NN	O	I-protein
molecule	NN	O	I-protein
that	NN	O	O
facilitates	NN	O	O
antigen	NN	O	O
processing	NN	O	O
by	NN	O	O
catalyzing	NN	O	O
the	NN	O	O
exchange	NN	O	O
of	NN	O	O
invariant	NN	O	O
chain-derived	NN	O	O
peptides	NN	O	O
(	NN	O	O
CLIP	NN	O	O
)	NN	O	O
from	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
for	NN	O	O
antigenic	NN	O	O
peptides	NN	O	O
.	NN	O	O

HLA-DO	NN	O	B-protein
is	NN	O	O
a	NN	O	O
second	NN	O	O
class	NN	O	B-protein
II-like	NN	O	I-protein
molecule	NN	O	I-protein
that	NN	O	O
physically	NN	O	O
associates	NN	O	O
with	NN	O	O
HLA-DM	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

HLA-DO	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
block	NN	O	O
HLA-DM	NN	O	B-protein
function	NN	O	O
.	NN	O	O

Purified	NN	O	O
HLA-DM-DO	NN	O	B-protein
complexes	NN	O	I-protein
could	NN	O	O
not	NN	O	O
promote	NN	O	O
peptide	NN	O	O
exchange	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
HLA-DO	NN	O	B-protein
in	NN	O	O
a	NN	O	O
class	NN	O	B-cell_line
II+	NN	O	I-cell_line
and	NN	O	I-cell_line
DM+	NN	O	I-cell_line
,	NN	O	I-cell_line
DO-	NN	O	I-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
caused	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
class	NN	O	B-protein
II-CLIP	NN	O	I-protein
complexes	NN	O	I-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
HLA-DO	NN	O	B-protein
blocked	NN	O	O
DM	NN	O	O
function	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
suggesting	NN	O	O
that	NN	O	O
HLA-DO	NN	O	B-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
modulator	NN	O	O
of	NN	O	O
class	NN	O	O
II-restricted	NN	O	O
antigen	NN	O	O
processing	NN	O	O
.	NN	O	O

-DOCSTART-	O

Vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
:	NN	O	O
no	NN	O	O
evidence	NN	O	O
for	NN	O	O
allele-specific	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
in	NN	O	O
cells	NN	O	O
which	NN	O	O
are	NN	O	O
heterozygous	NN	O	O
for	NN	O	O
the	NN	O	O
Taq	NN	O	B-protein
I	NN	O	I-protein
restriction	NN	O	I-protein
enzyme	NN	O	I-protein
polymorphism	NN	O	O
.	NN	O	O

Allelic	NN	O	O
variations	NN	O	O
of	NN	O	O
the	NN	O	O
vitamin	NN	O	B-DNA
D	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
VDR	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
risk	NN	O	O
of	NN	O	O
developing	NN	O	O
prostate	NN	O	O
cancer	NN	O	O
in	NN	O	O
men	NN	O	O
and	NN	O	O
osteoporosis	NN	O	O
in	NN	O	O
postmenopausal	NN	O	O
women	NN	O	O
.	NN	O	O

Three	NN	O	O
RFLPs	NN	O	B-DNA
(	NN	O	O
TaqI	NN	O	B-DNA
,	NN	O	O
ApaI	NN	O	B-DNA
,	NN	O	O
BsmI	NN	O	B-DNA
)	NN	O	O
define	NN	O	O
two	NN	O	O
common	NN	O	O
haplotypes	NN	O	O
:	NN	O	O
BAt	NN	O	O
and	NN	O	O
baT	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
these	NN	O	O
polymorphisms	NN	O	B-DNA
change	NN	O	O
the	NN	O	O
translated	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Since	NN	O	O
sequence	NN	O	B-DNA
variations	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
UTR	NN	O	I-DNA
of	NN	O	O
VDR	NN	O	B-protein
have	NN	O	O
been	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
different	NN	O	O
haplotypes	NN	O	O
,	NN	O	O
investigators	NN	O	O
have	NN	O	O
proposed	NN	O	O
that	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
VDR	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
influenced	NN	O	O
by	NN	O	O
allelic	NN	O	O
variations	NN	O	O
.	NN	O	O

Indirect	NN	O	O
evidence	NN	O	O
suggested	NN	O	O
that	NN	O	O
allele	NN	O	B-DNA
T	NN	O	I-DNA
is	NN	O	O
less	NN	O	O
stable	NN	O	O
than	NN	O	O
allele	NN	O	B-DNA
t	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
a	NN	O	O
RT-PCR	NN	O	O
based	NN	O	O
approach	NN	O	O
to	NN	O	O
compare	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
the	NN	O	O
big	NN	O	B-DNA
T	NN	O	I-DNA
and	NN	O	I-DNA
small	NN	O	I-DNA
t	NN	O	I-DNA
allele	NN	O	I-DNA
in	NN	O	O
normal	NN	O	B-cell_type
heterozygous	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
heterozygous	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
NB4	NN	O	B-cell_line
(	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
)	NN	O	O
and	NN	O	O
PC-3	NN	O	B-cell_line
and	NN	O	O
DU	NN	O	B-cell_line
145	NN	O	I-cell_line
(	NN	O	O
prostate	NN	O	O
cancers	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
three	NN	O	O
cases	NN	O	O
,	NN	O	O
we	NN	O	O
did	NN	O	O
not	NN	O	O
find	NN	O	O
a	NN	O	O
significant	NN	O	O
difference	NN	O	O
in	NN	O	O
stability	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
we	NN	O	O
consistently	NN	O	O
observed	NN	O	O
30	NN	O	O
%	NN	O	O
less	NN	O	O
RT-PCR	NN	O	B-DNA
product	NN	O	I-DNA
derived	NN	O	O
from	NN	O	O
the	NN	O	O
small	NN	O	B-RNA
t	NN	O	I-RNA
allele	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
steady	NN	O	O
state	NN	O	O
,	NN	O	O
a	NN	O	O
finding	NN	O	O
which	NN	O	O
also	NN	O	O
speaks	NN	O	O
against	NN	O	O
a	NN	O	O
higher	NN	O	O
stability	NN	O	O
of	NN	O	O
the	NN	O	O
small	NN	O	B-RNA
t	NN	O	I-RNA
allele	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
a	NN	O	O
variation	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
rather	NN	O	O
than	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
between	NN	O	O
the	NN	O	O
alleles	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
an	NN	O	O
unknown	NN	O	O
gene	NN	O	O
or	NN	O	O
genes	NN	O	O
in	NN	O	O
linkage	NN	O	O
with	NN	O	O
the	NN	O	O
polymorphisms	NN	O	B-DNA
is	NN	O	O
(	NN	O	O
are	NN	O	O
)	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
risk	NN	O	O
of	NN	O	O
prostate	NN	O	O
cancer	NN	O	O
and	NN	O	O
VDR	NN	O	B-protein
polymorphisms	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Replication	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus-1	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
autocrine	NN	O	O
secretion	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
through	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
-alpha	NN	O	I-protein
controls	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
and	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
inducer	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
-1	NN	O	O
replication	NN	O	O
in	NN	O	O
chronically	NN	O	B-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
primary	NN	O	O
cultures	NN	O	O
of	NN	O	O
HIV-infected	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
using	NN	O	O
neutralizing	NN	O	O
anti-TNF-alpha	NN	O	B-protein
antibodies	NN	O	I-protein
or	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

Primary	NN	O	O
resting	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
produced	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
supported	NN	O	O
HIV	NN	O	O
replication	NN	O	O
after	NN	O	O
T-cell	NN	O	O
receptor	NN	O	O
activation	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
neutralizing	NN	O	O
anti-TNF-alpha	NN	O	B-protein
antibodies	NN	O	I-protein
drastically	NN	O	O
reduced	NN	O	O
p24	NN	O	B-protein
antigen	NN	O	I-protein
release	NN	O	O
and	NN	O	O
prevented	NN	O	O
CD4+	NN	O	B-protein
cell	NN	O	O
depletion	NN	O	O
associated	NN	O	O
with	NN	O	O
infection	NN	O	O
.	NN	O	O

Anti-TNF-alpha	NN	O	B-protein
also	NN	O	O
prevented	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
good	NN	O	O
correlation	NN	O	O
between	NN	O	O
this	NN	O	O
inhibition	NN	O	O
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
HIV	NN	O	O
replication	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
supplementing	NN	O	O
the	NN	O	O
cultures	NN	O	O
with	NN	O	O
high	NN	O	O
doses	NN	O	O
of	NN	O	O
IL-2	NN	O	O
reverted	NN	O	O
anti-	NN	O	O
TNF-alpha	NN	O	B-protein
inhibition	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
HIV	NN	O	O
p24	NN	O	B-protein
antigen	NN	O	I-protein
release	NN	O	O
or	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
cultures	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
anti-TNF-alpha	NN	O	B-protein
inhibited	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
-driven	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
activation	NN	O	O
,	NN	O	O
either	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-protein
Tat	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
support	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
for	NN	O	O
autocrine	NN	O	O
TNF-alpha	NN	O	B-protein
secretion	NN	O	O
in	NN	O	O
controlling	NN	O	O
HIV	NN	O	O
replication	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
because	NN	O	O
of	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
maintain	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
elevated	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Hypoxia	NN	O	O
enhances	NN	O	O
induction	NN	O	O
of	NN	O	O
endothelial	NN	O	B-protein
ICAM-1	NN	O	I-protein
:	NN	O	O
role	NN	O	O
for	NN	O	O
metabolic	NN	O	O
acidosis	NN	O	O
and	NN	O	O
proteasomes	NN	O	B-protein
.	NN	O	O

Intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
molecule	NN	O	O
in	NN	O	O
promotion	NN	O	O
of	NN	O	O
polymorphonuclear	NN	O	O
neutrophil	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
during	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Coincident	NN	O	O
with	NN	O	O
many	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
is	NN	O	O
tissue	NN	O	O
hypoxia	NN	O	O
.	NN	O	O

Thus	NN	O	O
we	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
combinations	NN	O	O
of	NN	O	O
hypoxia	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
stimuli	NN	O	O
may	NN	O	O
differentially	NN	O	O
regulate	NN	O	O
expression	NN	O	O
of	NN	O	O
endothelial	NN	O	B-protein
ICAM-1	NN	O	I-protein
.	NN	O	O

Human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
hypoxia	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
added	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
and	NN	O	O
examined	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
functional	NN	O	O
ICAM-1	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
hypoxia	NN	O	O
alone	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
LPS	NN	O	O
and	NN	O	O
hypoxia	NN	O	O
enhanced	NN	O	O
(	NN	O	O
3	NN	O	O
+/-	NN	O	O
0.4-fold	NN	O	O
over	NN	O	O
normoxia	NN	O	O
)	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Combinations	NN	O	O
of	NN	O	O
hypoxia	NN	O	O
and	NN	O	O
LPS	NN	O	O
significantly	NN	O	O
increased	NN	O	O
lymphocyte	NN	O	O
binding	NN	O	O
,	NN	O	O
and	NN	O	O
such	NN	O	O
increases	NN	O	O
were	NN	O	O
inhibited	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
anti-ICAM-1	NN	O	B-protein
antibodies	NN	O	I-protein
or	NN	O	O
antisense	NN	O	O
oligonucleotides	NN	O	O
.	NN	O	O

Hypoxic	NN	O	O
endothelia	NN	O	O
showed	NN	O	O
a	NN	O	O
>	NN	O	O
10-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
proteasome	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
and	NN	O	O
combinations	NN	O	O
of	NN	O	O
hypoxia	NN	O	O
and	NN	O	O
LPS	NN	O	O
enhanced	NN	O	O
proteasome	NN	O	B-protein
-dependent	NN	O	O
cytoplasmic-to-nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
p65	NN	O	I-protein
(	NN	O	I-protein
Rel	NN	O	I-protein
A	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
.	NN	O	O

Such	NN	O	O
proteasome	NN	O	B-protein
activation	NN	O	O
correlated	NN	O	O
with	NN	O	O
hypoxia-evoked	NN	O	O
decreases	NN	O	O
in	NN	O	O
both	NN	O	O
extracellular	NN	O	O
and	NN	O	O
intracellular	NN	O	O
pH	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
from	NN	O	O
these	NN	O	O
studies	NN	O	O
that	NN	O	O
endothelial	NN	O	O
hypoxia	NN	O	O
provides	NN	O	O
a	NN	O	O
novel	NN	O	O
,	NN	O	O
proteasome	NN	O	B-protein
-dependent	NN	O	O
stimulus	NN	O	O
for	NN	O	O
ICAM-1	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differanisole	NN	O	O
A	NN	O	O
,	NN	O	O
a	NN	O	O
novel	NN	O	O
antitumor	NN	O	O
antibiotic	NN	O	O
,	NN	O	O
enhances	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
induced	NN	O	O
by	NN	O	O
9-cis	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
.	NN	O	O

Differanisole	NN	O	O
A	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
5-dichloro-2-hydroxy-4-methoxy-6-n-propylbenzoic	NN	O	O
acid	NN	O	O
,	NN	O	O
inhibited	NN	O	O
growth	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
compound	NN	O	O
induced	NN	O	O
G1	NN	O	O
arrest	NN	O	O
and	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
the	NN	O	O
differentiation-inducing	NN	O	O
effect	NN	O	O
was	NN	O	O
modest	NN	O	O
.	NN	O	O

Differanisole	NN	O	O
A	NN	O	O
and	NN	O	O
9-cis	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
9cisRA	NN	O	O
)	NN	O	O
synergistically	NN	O	O
inhibited	NN	O	O
the	NN	O	O
growth	NN	O	O
and	NN	O	O
induced	NN	O	O
functional	NN	O	O
and	NN	O	O
morphologic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
and	NN	O	I-cell_line
NB4	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
combined	NN	O	O
treatment	NN	O	O
with	NN	O	O
differanisole	NN	O	O
A	NN	O	O
and	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
or	NN	O	O
1alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
was	NN	O	O
less	NN	O	O
effective	NN	O	O
.	NN	O	O

Similar	NN	O	O
results	NN	O	O
were	NN	O	O
obtained	NN	O	O
in	NN	O	O
primary	NN	O	O
culture	NN	O	O
of	NN	O	O
leukemia	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

The	NN	O	O
synergistic	NN	O	O
effect	NN	O	O
on	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
differentiation	NN	O	O
required	NN	O	O
simultaneous	NN	O	O
treatment	NN	O	O
with	NN	O	O
differanisole	NN	O	O
A	NN	O	O
and	NN	O	O
9cisRA	NN	O	O
.	NN	O	O

Differanisole	NN	O	O
A	NN	O	O
and	NN	O	O
an	NN	O	O
RXR-specific	NN	O	O
ligand	NN	O	O
(	NN	O	O
Ro47-5944	NN	O	O
)	NN	O	O
cooperatively	NN	O	O
inhibited	NN	O	O
the	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
combined	NN	O	O
effect	NN	O	O
of	NN	O	O
differanisole	NN	O	O
A	NN	O	O
and	NN	O	O
an	NN	O	O
RAR-specific	NN	O	O
ligand	NN	O	O
Am80	NN	O	O
was	NN	O	O
just	NN	O	O
additive	NN	O	O
.	NN	O	O

Differanisole	NN	O	O
A	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
9cisRA	NN	O	O
may	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
therapy	NN	O	O
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
JAK2	NN	O	B-protein
/STAT5	NN	O	B-protein
pathway	NN	O	O
is	NN	O	O
commonly	NN	O	O
involved	NN	O	O
in	NN	O	O
signaling	NN	O	O
through	NN	O	O
the	NN	O	O
human	NN	O	B-protein
IL-5	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
JAK	NN	O	B-protein
(	NN	O	I-protein
Janus	NN	O	I-protein
kinase	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
and	NN	O	O
the	NN	O	O
STATs	NN	O	B-protein
(	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
)	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
evaluated	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
JAK	NN	O	B-protein
/STAT	NN	O	B-protein
pathway	NN	O	O
upon	NN	O	O
human	NN	O	B-protein
interleukin-5	NN	O	I-protein
(	NN	O	O
hIL-5	NN	O	B-protein
)	NN	O	O
stimulation	NN	O	O
of	NN	O	O
two	NN	O	O
different	NN	O	O
hIL-5-responsive	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
hIL-5	NN	O	B-cell_line
receptor	NN	O	I-cell_line
alpha-subunit	NN	O	I-cell_line
(	NN	O	I-cell_line
hIL-5R	NN	O	I-cell_line
alpha	NN	O	I-cell_line
)	NN	O	I-cell_line
cDNA-transfected	NN	O	I-cell_line
TF-1	NN	O	I-cell_line
(	NN	O	O
TF-h5R	NN	O	B-cell_line
alpha	NN	O	I-cell_line
)	NN	O	O
and	NN	O	O
butyric-acid-treated	NN	O	B-cell_line
YY-1	NN	O	I-cell_line
(	NN	O	O
YY-Bu	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
and	NN	O	O
peripheral	NN	O	B-cell_line
eosinophils	NN	O	I-cell_line
.	NN	O	O

Immunoprecipitation	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
JAK2	NN	O	B-protein
and	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
were	NN	O	O
induced	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
hIL-5	NN	O	B-protein
in	NN	O	O
all	NN	O	O
three	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
while	NN	O	O
STAT1	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
only	NN	O	O
observed	NN	O	O
in	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
JAK2	NN	O	B-protein
/STAT5	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
a	NN	O	O
common	NN	O	O
JAK	NN	O	B-protein
/STAT	NN	O	B-protein
pathway	NN	O	O
for	NN	O	O
hIL-5	NN	O	B-protein
-mediated	NN	O	O
signal	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Dysregulation	NN	O	O
of	NN	O	O
monocytic	NN	O	B-protein
nuclear	NN	O	I-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
by	NN	O	O
oxidized	NN	O	O
low-density	NN	O	B-protein
lipoprotein	NN	O	I-protein
.	NN	O	O

Nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
/Rel	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
atherosclerosis	NN	O	O
,	NN	O	O
as	NN	O	O
is	NN	O	O
suggested	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
human	NN	O	O
atherosclerotic	NN	O	O
lesions	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
oxidized	NN	O	B-protein
LDL	NN	O	I-protein
(	NN	O	O
oxLDL	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
system	NN	O	O
in	NN	O	O
human	NN	O	O
THP-1	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
as	NN	O	O
adherent	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
short-term	NN	O	O
incubation	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
with	NN	O	O
oxLDL	NN	O	B-protein
activated	NN	O	I-protein
p50/p65	NN	O	I-protein
containing	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
dimers	NN	O	I-protein
and	NN	O	O
induced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
target	NN	O	B-DNA
gene	NN	O	I-DNA
IL-8	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
and	NN	O	O
H2O2	NN	O	O
scavenger	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
and	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
inhibitor	NN	O	O
PSI	NN	O	O
.	NN	O	O

The	NN	O	O
oxLDL	NN	O	B-protein
-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
initial	NN	O	O
depletion	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
followed	NN	O	O
by	NN	O	O
a	NN	O	O
slight	NN	O	O
transient	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
this	NN	O	O
inhibitor	NN	O	O
protein	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
long-term	NN	O	O
treatment	NN	O	O
with	NN	O	O
oxLDL	NN	O	B-protein
prevented	NN	O	O
the	NN	O	O
lipopolysaccharide-induced	NN	O	O
depletion	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
,	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
inhibition	NN	O	O
of	NN	O	O
both	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor-alpha	NN	O	I-DNA
and	NN	O	I-DNA
interleukin-1	NN	O	I-DNA
beta	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
observations	NN	O	O
provide	NN	O	O
additional	NN	O	O
evidence	NN	O	O
that	NN	O	O
oxLDL	NN	O	B-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
modulator	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
(	NN	O	O
dys	NN	O	O
)	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
is	NN	O	O
likely	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
atherogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
CD18	NN	O	B-protein
-mediated	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
the	NN	O	O
corneal	NN	O	O
endothelium	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

PURPOSE	NN	O	O
:	NN	O	O
To	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
cultured	NN	O	B-cell_line
corneal	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
CD18	NN	O	B-protein
antigen	NN	O	I-protein
,	NN	O	O
and	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
dexamethasone	NN	O	O
directly	NN	O	O
suppresses	NN	O	O
adhesion	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NFkappaB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Cultured	NN	O	O
bovine	NN	O	O
corneal	NN	O	O
endothelium	NN	O	O
was	NN	O	O
stimulated	NN	O	O
for	NN	O	O
6	NN	O	O
hours	NN	O	O
by	NN	O	O
40	NN	O	O
micron/ml	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNFalpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
was	NN	O	O
added	NN	O	O
1	NN	O	O
hour	NN	O	O
before	NN	O	O
TNFalpha	NN	O	B-protein
stimulation	NN	O	O
in	NN	O	O
the	NN	O	O
dexamethasone	NN	O	O
group	NN	O	O
.	NN	O	O

After	NN	O	O
stimulation	NN	O	O
,	NN	O	O
neutrophils	NN	O	O
separated	NN	O	O
from	NN	O	O
a	NN	O	O
healthy	NN	O	O
human	NN	O	O
volunteer	NN	O	O
were	NN	O	O
added	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
anti-CD18	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

The	NN	O	O
culture	NN	O	O
plate	NN	O	O
was	NN	O	O
settled	NN	O	O
for	NN	O	O
15	NN	O	O
minutes	NN	O	O
at	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
,	NN	O	O
and	NN	O	O
then	NN	O	O
neutrophils	NN	O	B-cell_type
were	NN	O	O
activated	NN	O	O
by	NN	O	O
N-formyl-methionyl-leucyl-phenylalanine	NN	O	O
for	NN	O	O
5	NN	O	O
minutes	NN	O	O
.	NN	O	O

Nonadherent	NN	O	O
neutrophils	NN	O	B-cell_type
were	NN	O	O
removed	NN	O	O
by	NN	O	O
sealing	NN	O	O
and	NN	O	O
inverting	NN	O	O
the	NN	O	O
culture	NN	O	O
well	NN	O	O
.	NN	O	O

The	NN	O	O
intracellular	NN	O	O
localization	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
after	NN	O	O
TNFalpha	NN	O	B-protein
simulation	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
confocal	NN	O	O
immunocytochemistry	NN	O	O
using	NN	O	O
an	NN	O	O
anti-p65	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Neutrophil	NN	O	O
adhesion	NN	O	O
to	NN	O	O
cultured	NN	O	B-cell_line
corneal	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
increased	NN	O	O
significantly	NN	O	O
on	NN	O	O
exposure	NN	O	O
to	NN	O	O
TNFalpha	NN	O	B-protein
(	NN	O	O
451.4+/-45.4	NN	O	O
cells/mm2	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
16	NN	O	O
)	NN	O	O
compared	NN	O	O
to	NN	O	O
control	NN	O	O
(	NN	O	O
156.7+/-27.3	NN	O	O
cells/mm2	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
16	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
increased	NN	O	O
adhesion	NN	O	O
was	NN	O	O
suppressed	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
anti-CD18	NN	O	B-protein
antibody	NN	O	I-protein
(	NN	O	O
157.6+/-25.1	NN	O	O
cells/mm2	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
8	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
and	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
M	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
207.9+/-31.5	NN	O	O
cells/mm2	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
10	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

Immunocytochemistry	NN	O	O
60	NN	O	O
minutes	NN	O	O
after	NN	O	O
stimulation	NN	O	O
revealed	NN	O	O
that	NN	O	O
NFkappaB	NN	O	B-protein
was	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
in	NN	O	O
unstimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
TNFalpha	NN	O	B-protein
caused	NN	O	O
NFkappaB	NN	O	B-protein
to	NN	O	O
translocate	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
tapered	NN	O	O
NFkappaB	NN	O	B-protein
translocation	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Leukocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
the	NN	O	O
corneal	NN	O	O
endothelium	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
CD18	NN	O	B-protein
expressed	NN	O	O
on	NN	O	O
activated	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
the	NN	O	O
endothelium	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
inhibited	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
;	NN	O	O
this	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
in	NN	O	O
part	NN	O	O
to	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
entry	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
characterization	NN	O	O
and	NN	O	O
pattern	NN	O	O
of	NN	O	O
tissue	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
for	NN	O	O
neutrophil	NN	O	B-protein
gelatinase-associated	NN	O	I-protein
lipocalin	NN	O	I-protein
from	NN	O	O
humans	NN	O	O
.	NN	O	O

Neutrophil	NN	O	B-protein
gelatinase-associated	NN	O	I-protein
lipocalin	NN	O	I-protein
(	NN	O	O
NGAL	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
25-kDa	NN	O	B-protein
lipocalin	NN	O	I-protein
first	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
protein	NN	O	O
stored	NN	O	O
in	NN	O	O
specific	NN	O	O
granules	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_type
neutrophil	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
protein	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
bind	NN	O	O
small	NN	O	O
lipophilic	NN	O	O
substances	NN	O	O
such	NN	O	O
as	NN	O	O
bacterial	NN	O	O
derived	NN	O	O
formylpeptides	NN	O	O
and	NN	O	O
lipopolysaccharides	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
and	NN	O	O
might	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
modulator	NN	O	O
of	NN	O	O
inflammation	NN	O	O
.	NN	O	O

To	NN	O	O
characterize	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
NGAL	NN	O	B-protein
further	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
a	NN	O	O
5869-bp	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
NGAL	NN	O	B-DNA
gene	NN	O	I-DNA
including	NN	O	O
1695	NN	O	O
bp	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
nontranscribed	NN	O	I-DNA
region	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
3696-bp	NN	O	B-DNA
coding	NN	O	I-DNA
region	NN	O	I-DNA
encompassing	NN	O	O
seven	NN	O	B-DNA
exons	NN	O	I-DNA
and	NN	O	I-DNA
six	NN	O	I-DNA
introns	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	B-DNA
start	NN	O	I-DNA
sites	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
by	NN	O	O
an	NN	O	O
RNase	NN	O	B-protein
protection	NN	O	O
assay	NN	O	O
.	NN	O	O

The	NN	O	O
NGAL	NN	O	B-protein
gene	NN	O	O
is	NN	O	O
highly	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
gene	NN	O	I-DNA
24p3	NN	O	I-DNA
.	NN	O	O

NGAL	NN	O	B-protein
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
bone	NN	O	O
marrow	NN	O	O
and	NN	O	O
in	NN	O	O
tissues	NN	O	O
that	NN	O	O
are	NN	O	O
prone	NN	O	O
to	NN	O	O
exposure	NN	O	O
to	NN	O	O
microorganisms	NN	O	O
.	NN	O	O

Potential	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
NGAL	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
computer	NN	O	O
analysis	NN	O	O
and	NN	O	O
include	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
CTF/CBP	NN	O	B-protein
,	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
GATA-1	NN	O	B-protein
and	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
LPS-inducible	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
express	NN	O	O
GH-N	NN	O	B-RNA
gene	NN	O	I-RNA
transcripts	NN	O	I-RNA
and	NN	O	O
the	NN	O	O
pituitary	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Pit-1b	NN	O	B-protein
.	NN	O	O

Since	NN	O	O
GH	NN	O	B-protein
stimulates	NN	O	O
the	NN	O	O
development	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
granulocytes	NN	O	B-cell_type
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
GH	NN	O	B-protein
in	NN	O	O
granulocyte	NN	O	B-cell_type
subsets	NN	O	I-cell_type
.	NN	O	O

By	NN	O	O
immunocytochemistry	NN	O	O
,	NN	O	O
25	NN	O	O
+/-	NN	O	O
7	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
were	NN	O	O
shown	NN	O	O
to	NN	O	O
express	NN	O	O
immunoreactive	NN	O	B-protein
GH	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
eosinophils	NN	O	B-cell_type
were	NN	O	O
negative	NN	O	O
.	NN	O	O

Reversed	NN	O	O
transcription	NN	O	O
(	NN	O	O
RT	NN	O	O
)	NN	O	O
-PCR	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
GH	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
neutrophils	NN	O	B-cell_type
.	NN	O	O

Restriction	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
neutrophils	NN	O	B-cell_type
express	NN	O	O
the	NN	O	O
GH-N	NN	O	B-DNA
gene	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
the	NN	O	O
GH-V	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
that	NN	O	O
neutrophils	NN	O	B-cell_type
express	NN	O	O
an	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
variant	NN	O	O
of	NN	O	O
the	NN	O	O
pituitary	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Pit-1	NN	O	B-protein
,	NN	O	O
designated	NN	O	O
Pit-1b	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
BHRF1	NN	O	B-protein
to	NN	O	O
inhibit	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
stimulus	NN	O	O
and	NN	O	O
cell	NN	O	O
type	NN	O	O
.	NN	O	O

The	NN	O	O
development	NN	O	O
of	NN	O	O
resistance	NN	O	O
to	NN	O	O
host	NN	O	O
defense	NN	O	O
mechanisms	NN	O	O
such	NN	O	O
as	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor	NN	O	O
(	NN	O	O
TNF	NN	O	O
)	NN	O	O
-	NN	O	O
and	NN	O	O
Fas-mediated	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
transformed	NN	O	B-cell_type
or	NN	O	O
virus-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
a	NN	O	O
critical	NN	O	O
component	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
disease	NN	O	O
.	NN	O	O

To	NN	O	O
find	NN	O	O
genes	NN	O	O
that	NN	O	O
protect	NN	O	O
cells	NN	O	O
from	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
an	NN	O	O
expression	NN	O	O
cloning	NN	O	O
strategy	NN	O	O
and	NN	O	O
identified	NN	O	O
BHRF1	NN	O	B-protein
,	NN	O	O
an	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
(	NN	O	I-protein
EBV	NN	O	I-protein
)	NN	O	I-protein
early-lytic-cycle	NN	O	I-protein
protein	NN	O	I-protein
with	NN	O	O
distant	NN	O	O
homology	NN	O	O
to	NN	O	O
Bcl-2	NN	O	B-protein
,	NN	O	O
as	NN	O	O
an	NN	O	O
anti-apoptosis	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
BHRF1	NN	O	B-protein
in	NN	O	O
MCF-Fas	NN	O	B-cell_line
cells	NN	O	I-cell_line
conferred	NN	O	O
nearly	NN	O	O
complete	NN	O	O
resistance	NN	O	O
against	NN	O	O
both	NN	O	O
anti-Fas	NN	O	B-protein
antibody	NN	O	I-protein
and	NN	O	O
TNF-mediated	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
BHRF1	NN	O	B-protein
protected	NN	O	O
these	NN	O	O
cells	NN	O	O
from	NN	O	O
monocyte-mediated	NN	O	O
killing	NN	O	O
but	NN	O	O
failed	NN	O	O
to	NN	O	O
protect	NN	O	O
them	NN	O	O
from	NN	O	O
killing	NN	O	O
mediated	NN	O	O
by	NN	O	O
lymphokine-activated	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
BHRF1	NN	O	B-protein
to	NN	O	O
protect	NN	O	O
MCF-Fas	NN	O	B-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
various	NN	O	O
stimuli	NN	O	O
was	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
and	NN	O	O
Bcl-xL	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
BHRF1	NN	O	B-protein
resembled	NN	O	O
that	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
and	NN	O	O
Bcl-xL	NN	O	B-protein
as	NN	O	O
it	NN	O	O
inhibited	NN	O	O
TNF-	NN	O	O
and	NN	O	O
anti-Fas-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
two	NN	O	O
enzymes	NN	O	O
participating	NN	O	O
in	NN	O	O
the	NN	O	O
apoptosis	NN	O	O
pathway	NN	O	O
,	NN	O	O
cytosolic	NN	O	B-protein
phospholipase	NN	O	I-protein
A2	NN	O	I-protein
and	NN	O	O
caspase-3/CPP32	NN	O	B-protein
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
interfere	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB-like	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

A	NN	O	O
putative	NN	O	O
function	NN	O	O
of	NN	O	O
BHRF1	NN	O	B-protein
in	NN	O	O
EBV-infected	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
to	NN	O	O
protect	NN	O	O
virus-infected	NN	O	O
cells	NN	O	O
from	NN	O	O
TNF-	NN	O	O
and/or	NN	O	O
anti-Fas-	NN	O	O
induced	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
maximize	NN	O	O
virus	NN	O	O
production	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
neither	NN	O	O
BHRF1	NN	O	B-protein
nor	NN	O	O
Bcl-2	NN	O	B-protein
in	NN	O	O
a	NN	O	O
B-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
BJAB	NN	O	B-cell_line
,	NN	O	O
protected	NN	O	O
the	NN	O	O
cells	NN	O	O
from	NN	O	O
anti-Fas-mediated	NN	O	O
apoptosis	NN	O	O
even	NN	O	O
though	NN	O	O
they	NN	O	O
increased	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
serum-starved	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
protective	NN	O	O
role	NN	O	O
of	NN	O	O
BHRF1	NN	O	B-protein
against	NN	O	O
apoptosis	NN	O	O
resembles	NN	O	O
that	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
in	NN	O	O
being	NN	O	O
cell	NN	O	O
type	NN	O	O
specific	NN	O	O
and	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
stimulus	NN	O	O
.	NN	O	O

-DOCSTART-	O

Genomic	NN	O	O
organization	NN	O	O
,	NN	O	O
sequence	NN	O	O
,	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
eotaxin	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Eotaxin	NN	O	B-protein
is	NN	O	O
an	NN	O	O
eosinophil	NN	O	B-protein
specific	NN	O	I-protein
beta-chemokine	NN	O	I-protein
assumed	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
eosinophilic	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
such	NN	O	O
as	NN	O	O
atopic	NN	O	O
dermatitis	NN	O	O
,	NN	O	O
allergic	NN	O	O
rhinitis	NN	O	O
,	NN	O	O
asthma	NN	O	O
and	NN	O	O
parasitic	NN	O	O
infections	NN	O	O
.	NN	O	O

Its	NN	O	O
expression	NN	O	O
is	NN	O	O
stimulus-	NN	O	O
and	NN	O	O
cell-specific	NN	O	O
.	NN	O	O

We	NN	O	O
here	NN	O	O
describe	NN	O	O
the	NN	O	O
genomic	NN	O	O
organisation	NN	O	O
(	NN	O	O
3	NN	O	O
exons	NN	O	B-DNA
of	NN	O	O
132	NN	O	O
,	NN	O	O
112	NN	O	O
and	NN	O	O
542	NN	O	O
bp	NN	O	O
and	NN	O	O
2	NN	O	O
introns	NN	O	B-DNA
of	NN	O	O
1211	NN	O	O
and	NN	O	O
378	NN	O	O
bp	NN	O	O
)	NN	O	O
and	NN	O	O
sequence	NN	O	O
including	NN	O	O
3	NN	O	O
kb	NN	O	O
of	NN	O	O
DNA	NN	O	O
from	NN	O	O
the	NN	O	O
immediate	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
upstream	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
eotaxin	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Among	NN	O	O
the	NN	O	O
regulatory	NN	O	B-DNA
promoter	NN	O	I-DNA
elements	NN	O	I-DNA
potentially	NN	O	O
regulating	NN	O	O
eotaxin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
and/or	NN	O	O
mediating	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
anti-inflammatory	NN	O	O
drugs	NN	O	O
we	NN	O	O
identified	NN	O	O
consensus	NN	O	B-DNA
sequences	NN	O	I-DNA
known	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
like	NN	O	O
NF-IL6	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
NF-kappa-B	NN	O	B-DNA
like	NN	O	I-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
and	NN	O	O
gamma-interferon-	NN	O	B-DNA
as	NN	O	I-DNA
well	NN	O	I-DNA
as	NN	O	I-DNA
glucocorticoid	NN	O	I-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
interaction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
with	NN	O	O
the	NN	O	O
leukocyte-specific	NN	O	B-DNA
pp52	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
leukocyte-specific	NN	O	O
,	NN	O	O
cytoskeleton-binding	NN	O	O
pp52	NN	O	O
(	NN	O	O
LSP-1	NN	O	O
,	NN	O	O
WP-34	NN	O	O
)	NN	O	O
protein	NN	O	O
is	NN	O	O
widely	NN	O	O
expressed	NN	O	O
in	NN	O	O
multiple	NN	O	O
leukocyte	NN	O	O
lineages	NN	O	O
,	NN	O	O
including	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
granulocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
previously	NN	O	O
detected	NN	O	O
a	NN	O	O
tissue-specific	NN	O	B-DNA
promoter	NN	O	I-DNA
preceding	NN	O	O
the	NN	O	O
exon	NN	O	O
encoding	NN	O	O
the	NN	O	O
N	NN	O	B-protein
terminus	NN	O	I-protein
of	NN	O	O
the	NN	O	O
pp52	NN	O	B-protein
leukocyte	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
functional	NN	O	O
characterization	NN	O	O
of	NN	O	O
this	NN	O	O
promoter	NN	O	O
and	NN	O	O
identification	NN	O	O
of	NN	O	O
the	NN	O	O
factors	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
regulate	NN	O	O
its	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
pp52	NN	O	B-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
an	NN	O	O
initiator	NN	O	O
specifying	NN	O	O
the	NN	O	O
unique	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
terminus	NN	O	I-DNA
of	NN	O	O
pp52	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
tandem	NN	O	O
pairs	NN	O	O
of	NN	O	O
Ets	NN	O	B-DNA
and	NN	O	I-DNA
SP1	NN	O	I-DNA
motifs	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
lone	NN	O	B-DNA
C/EBP	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

All	NN	O	O
these	NN	O	O
motifs	NN	O	O
are	NN	O	O
essential	NN	O	O
and	NN	O	O
collectively	NN	O	O
control	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

DNA	NN	O	O
binding	NN	O	O
studies	NN	O	O
and	NN	O	O
Ab	NN	O	O
supershift	NN	O	O
assays	NN	O	O
revealed	NN	O	O
that	NN	O	O
different	NN	O	O
combinations	NN	O	O
of	NN	O	O
factors	NN	O	O
interact	NN	O	O
with	NN	O	O
these	NN	O	O
motifs	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
vs	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
Ets	NN	O	B-DNA
motifs	NN	O	I-DNA
are	NN	O	O
preferentially	NN	O	O
bound	NN	O	O
by	NN	O	O
PU-1	NN	O	B-protein
in	NN	O	O
B	NN	O	O
cell	NN	O	O
extracts	NN	O	O
from	NN	O	O
all	NN	O	O
stages	NN	O	O
of	NN	O	O
development	NN	O	O
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
different	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
member	NN	O	I-protein
reacts	NN	O	O
with	NN	O	O
these	NN	O	O
motifs	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
extracts	NN	O	O
.	NN	O	O

The	NN	O	O
C/EBP	NN	O	B-DNA
motif	NN	O	I-DNA
is	NN	O	O
bound	NN	O	O
by	NN	O	O
Ig/EBP-1	NN	O	B-protein
in	NN	O	O
pre-B	NN	O	O
cell	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
extracts	NN	O	O
,	NN	O	O
but	NN	O	O
is	NN	O	O
replaced	NN	O	O
by	NN	O	O
nuclear	NN	O	B-protein
factor-IL-6beta	NN	O	I-protein
or	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor-IL-6beta-Ig/EBP-1	NN	O	I-protein
heterodimer	NN	O	I-protein
in	NN	O	O
plasmacytoma	NN	O	O
cell	NN	O	O
extracts	NN	O	O
.	NN	O	O

Despite	NN	O	O
its	NN	O	O
reported	NN	O	O
role	NN	O	O
as	NN	O	O
a	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
transcription	NN	O	O
,	NN	O	O
Ig/EBP-1	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
exert	NN	O	O
a	NN	O	O
stimulatory	NN	O	O
effect	NN	O	O
on	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

These	NN	O	O
findings	NN	O	O
reveal	NN	O	O
the	NN	O	O
features	NN	O	O
controlling	NN	O	O
the	NN	O	O
pp52	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
provide	NN	O	O
the	NN	O	O
foundation	NN	O	O
for	NN	O	O
determining	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
in	NN	O	O
other	NN	O	O
leukocyte	NN	O	B-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
B-cell	NN	O	O
commitment	NN	O	O
to	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
to	NN	O	O
memory	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

During	NN	O	O
humoral	NN	O	O
immune	NN	O	O
responses	NN	O	O
,	NN	O	O
B-lymphocyte	NN	O	O
activation	NN	O	O
is	NN	O	O
followed	NN	O	O
by	NN	O	O
differentiation	NN	O	O
along	NN	O	O
either	NN	O	O
the	NN	O	O
plasma	NN	O	O
cell	NN	O	O
pathway	NN	O	O
or	NN	O	O
the	NN	O	O
memory	NN	O	O
B-cell	NN	O	O
pathway	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
CD40-CD40	NN	O	B-protein
ligand	NN	O	I-protein
,	NN	O	O
OX-OX40	NN	O	B-protein
ligand	NN	O	I-protein
,	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
intracellular	NN	O	O
transcriptional	NN	O	O
factors	NN	O	O
may	NN	O	O
all	NN	O	O
contribute	NN	O	O
to	NN	O	O
B-lymphocyte	NN	O	O
differentiation	NN	O	O
control	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
sites	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
transcription	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
are	NN	O	O
important	NN	O	O
for	NN	O	O
virus	NN	O	O
infectivity	NN	O	O
.	NN	O	O

When	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
,	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
promoter	NN	O	B-DNA
contains	NN	O	O
a	NN	O	O
nucleosome-free	NN	O	O
region	NN	O	O
encompassing	NN	O	O
both	NN	O	O
the	NN	O	O
promoter/enhancer	NN	O	B-DNA
region	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
large	NN	O	O
region	NN	O	O
(	NN	O	O
255	NN	O	B-DNA
nucleotides	NN	O	I-DNA
[	NN	O	O
nt	NN	O	O
]	NN	O	O
)	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
identified	NN	O	O
new	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
nt	NN	O	B-DNA
465	NN	O	I-DNA
to	NN	O	I-DNA
720	NN	O	I-DNA
)	NN	O	O
:	NN	O	O
three	NN	O	O
AP-1	NN	O	B-DNA
sites	NN	O	I-DNA
(	NN	O	O
I	NN	O	O
,	NN	O	O
II	NN	O	O
,	NN	O	O
and	NN	O	O
III	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
AP3-like	NN	O	B-DNA
motif	NN	O	I-DNA
(	NN	O	O
AP3-L	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
a	NN	O	O
downstream	NN	O	B-DNA
binding	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
DBF	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
juxtaposed	NN	O	B-DNA
Sp1	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
DBF	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
an	NN	O	O
interferon-responsive	NN	O	B-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
IRF	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
the	NN	O	O
AP3-L	NN	O	B-DNA
motif	NN	O	I-DNA
binds	NN	O	O
the	NN	O	O
T-cell-specific	NN	O	B-protein
factor	NN	O	I-protein
NF-AT	NN	O	B-protein
.	NN	O	O

Mutations	NN	O	O
that	NN	O	O
abolish	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
each	NN	O	O
factor	NN	O	O
to	NN	O	O
its	NN	O	O
cognate	NN	O	O
site	NN	O	O
are	NN	O	O
introduced	NN	O	O
in	NN	O	O
an	NN	O	O
infectious	NN	O	O
HIV-1	NN	O	O
molecular	NN	O	O
clone	NN	O	O
to	NN	O	O
study	NN	O	O
their	NN	O	O
effect	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
and	NN	O	O
replication	NN	O	O
.	NN	O	O

Individual	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
DBF	NN	O	O
or	NN	O	O
AP3-L	NN	O	B-DNA
site	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
double	NN	O	O
mutation	NN	O	B-DNA
AP-1	NN	O	I-DNA
(	NN	O	I-DNA
III	NN	O	I-DNA
)	NN	O	I-DNA
/AP3-L	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
affect	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
wild-type	NN	O	O
virus	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
proviruses	NN	O	O
carrying	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Sp1	NN	O	B-DNA
sites	NN	O	I-DNA
were	NN	O	O
totally	NN	O	O
defective	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
replication	NN	O	O
.	NN	O	O

Virus	NN	O	O
production	NN	O	O
occurred	NN	O	O
with	NN	O	O
slightly	NN	O	O
delayed	NN	O	O
kinetics	NN	O	O
for	NN	O	O
viruses	NN	O	O
containing	NN	O	O
combined	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
(	NN	O	I-DNA
III	NN	O	I-DNA
)	NN	O	I-DNA
,	NN	O	O
AP3-L	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
DBF	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
in	NN	O	O
the	NN	O	O
AP3-L	NN	O	B-DNA
and	NN	O	O
DBF-sites	NN	O	B-DNA
,	NN	O	O
whereas	NN	O	O
viruses	NN	O	O
mutated	NN	O	O
in	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
(	NN	O	I-DNA
I	NN	O	I-DNA
,	NN	O	I-DNA
II	NN	O	I-DNA
,	NN	O	I-DNA
III	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
AP3-L	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
in	NN	O	O
the	NN	O	O
AP-1	NN	O	O
(	NN	O	O
I	NN	O	O
,	NN	O	O
II	NN	O	O
,	NN	O	O
III	NN	O	O
)	NN	O	O
,	NN	O	O
AP3-L	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
DBF	NN	O	B-DNA
sites	NN	O	I-DNA
exhibited	NN	O	O
a	NN	O	O
severely	NN	O	O
defective	NN	O	O
replicative	NN	O	O
phenotype	NN	O	O
.	NN	O	O

No	NN	O	O
RNA-packaging	NN	O	O
defect	NN	O	O
could	NN	O	O
be	NN	O	O
measured	NN	O	O
for	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
viruses	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
quantification	NN	O	O
of	NN	O	O
their	NN	O	O
HIV	NN	O	B-RNA
genomic	NN	O	I-RNA
RNA	NN	O	I-RNA
.	NN	O	O

Measurement	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
promoter	NN	O	I-DNA
after	NN	O	O
transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
provirus	NN	O	I-DNA
DNA	NN	O	I-DNA
or	NN	O	O
of	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat-luciferase	NN	O	I-DNA
constructs	NN	O	I-DNA
showed	NN	O	O
a	NN	O	O
positive	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
the	NN	O	O
replication	NN	O	O
defects	NN	O	O
for	NN	O	O
most	NN	O	O
mutants	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
regulates	NN	O	O
expression	NN	O	O
of	NN	O	O
L-selectin	NN	O	B-protein
and	NN	O	O
CD11/CD18	NN	O	B-protein
on	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
raised	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
could	NN	O	O
diminish	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
direct	NN	O	O
leukocyte	NN	O	O
traffic	NN	O	O
into	NN	O	O
inflamed	NN	O	O
tissues	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
endothelial-leukocyte	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
and	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
investigate	NN	O	O
whether	NN	O	O
glucocorticoids	NN	O	O
also	NN	O	O
regulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
L-selectin	NN	O	B-protein
and	NN	O	O
CD11/CD18	NN	O	B-protein
integrins	NN	O	B-protein
on	NN	O	O
human	NN	O	B-cell_type
neutrophil	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
.	NN	O	O

METHODS	NN	O	O
AND	NN	O	O
RESULTS	NN	O	O
:	NN	O	O
Incubation	NN	O	O
of	NN	O	O
human	NN	O	O
whole	NN	O	O
blood	NN	O	O
with	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
PAF	NN	O	B-protein
,	NN	O	O
1	NN	O	O
mumol/L	NN	O	O
)	NN	O	O
evoked	NN	O	O
downregulation	NN	O	O
of	NN	O	O
L-selectin	NN	O	B-protein
and	NN	O	O
upregulation	NN	O	O
of	NN	O	O
CD11/CD18	NN	O	B-protein
adhesion	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
neutrophils	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

While	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
0.1	NN	O	O
nmol/L	NN	O	O
to	NN	O	O
100	NN	O	O
mumol/L	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
expression	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
on	NN	O	O
resting	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
,	NN	O	O
it	NN	O	O
attenuated	NN	O	O
the	NN	O	O
PAF	NN	O	B-protein
-induced	NN	O	O
changes	NN	O	O
in	NN	O	O
L-selectin	NN	O	B-protein
and	NN	O	O
CD18	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
a	NN	O	O
time-	NN	O	O
and	NN	O	O
concentration-dependent	NN	O	O
fashion	NN	O	O
with	NN	O	O
IC50	NN	O	O
values	NN	O	O
of	NN	O	O
31	NN	O	O
and	NN	O	O
13	NN	O	O
nmol/L	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

These	NN	O	O
effects	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
were	NN	O	O
completely	NN	O	O
aborted	NN	O	O
by	NN	O	O
RU-486	NN	O	O
(	NN	O	O
10	NN	O	O
mumol/L	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
blocks	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
inhibitor	NN	O	O
cycloheximide	NN	O	O
(	NN	O	O
35.5	NN	O	O
mumol/L	NN	O	O
)	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
,	NN	O	O
up	NN	O	O
to	NN	O	O
a	NN	O	O
concentration	NN	O	O
of	NN	O	O
1	NN	O	O
mumol/L	NN	O	O
,	NN	O	O
neither	NN	O	O
affected	NN	O	O
significantly	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
granule	NN	O	B-protein
enzymes	NN	O	I-protein
nor	NN	O	O
interfered	NN	O	O
with	NN	O	O
PAF	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
its	NN	O	O
membrane	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Our	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
at	NN	O	O
clinically	NN	O	O
relevant	NN	O	O
concentrations	NN	O	O
exert	NN	O	O
specific	NN	O	O
actions	NN	O	O
on	NN	O	O
expression	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
on	NN	O	O
activated	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
are	NN	O	O
mediated	NN	O	O
through	NN	O	O
ligation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
induction	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
novel	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
anti-inflammatory	NN	O	O
corticosteroids	NN	O	O
may	NN	O	O
inhibit	NN	O	O
leukocyte	NN	O	O
accumulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
immediate-early	NN	O	O
gene	NN	O	O
product	NN	O	O
Egr-1	NN	O	B-protein
regulates	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin-2	NN	O	I-DNA
receptor	NN	O	I-DNA
beta-chain	NN	O	I-DNA
promoter	NN	O	I-DNA
through	NN	O	O
noncanonical	NN	O	B-DNA
Egr	NN	O	I-DNA
and	NN	O	I-DNA
Sp1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
interleukin-2	NN	O	B-protein
IL-2	NN	O	I-protein
receptor	NN	O	I-protein
beta-chain	NN	O	I-protein
(	NN	O	O
IL-2Rbeta	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
essential	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
receptors	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-15	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
IL-2Rbeta	NN	O	B-protein
is	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
by	NN	O	O
lymphocytes	NN	O	O
,	NN	O	O
its	NN	O	O
expression	NN	O	O
can	NN	O	O
be	NN	O	O
further	NN	O	O
induced	NN	O	O
by	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
stimuli	NN	O	O
,	NN	O	O
including	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
characterized	NN	O	O
factors	NN	O	O
that	NN	O	O
bind	NN	O	O
to	NN	O	O
an	NN	O	O
enhancer	NN	O	O
region	NN	O	O
located	NN	O	O
between	NN	O	O
nucleotides	NN	O	B-DNA
-170	NN	O	I-DNA
and	NN	O	I-DNA
-139	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	O
IL-2Rbeta	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
Sp3	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
portion	NN	O	I-DNA
of	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
PMA-inducible	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
PIF	NN	O	B-protein
)	NN	O	O
mainly	NN	O	O
bound	NN	O	O
to	NN	O	O
its	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
portion	NN	O	I-DNA
and	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
Sp	NN	O	B-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
.	NN	O	O

In	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
induction	NN	O	O
of	NN	O	O
PIF	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
,	NN	O	O
required	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
sustained	NN	O	O
at	NN	O	O
a	NN	O	O
high	NN	O	O
level	NN	O	O
for	NN	O	O
at	NN	O	O
least	NN	O	O
23	NN	O	O
h	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
PIF	NN	O	B-protein
was	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
virus	NN	O	I-cell_line
type	NN	O	I-cell_line
1-transformed	NN	O	I-cell_line
MT-2	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
PIF	NN	O	B-protein
is	NN	O	O
Egr-1	NN	O	B-protein
based	NN	O	O
on	NN	O	O
its	NN	O	O
recognition	NN	O	O
by	NN	O	O
anti-	NN	O	O
Egr-1	NN	O	B-protein
antisera	NN	O	O
in	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
even	NN	O	O
though	NN	O	O
the	NN	O	O
IL-2Rbeta	NN	O	B-DNA
DNA	NN	O	I-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
differed	NN	O	O
substantially	NN	O	O
from	NN	O	O
the	NN	O	O
canonical	NN	O	O
Egr-1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
Egr-1	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
Sp	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
both	NN	O	O
sites	NN	O	O
were	NN	O	O
required	NN	O	O
for	NN	O	O
maximal	NN	O	O
IL-2Rbeta	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
HeLaS3	NN	O	O
cells	NN	O	O
,	NN	O	O
transfection	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
could	NN	O	O
drive	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	O
construct	NN	O	O
containing	NN	O	O
both	NN	O	O
sites	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
Egr-1	NN	O	B-protein
could	NN	O	O
form	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
kinetics	NN	O	O
that	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
upon	NN	O	O
PMA	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
Egr-1	NN	O	B-protein
physically	NN	O	O
and	NN	O	O
functionally	NN	O	O
cooperate	NN	O	O
to	NN	O	O
mediate	NN	O	O
maximal	NN	O	O
IL-2Rbeta	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Overexpression	NN	O	O
of	NN	O	O
HSF2-beta	NN	O	B-protein
inhibits	NN	O	O
hemin	NN	O	B-protein
-induced	NN	O	O
heat	NN	O	O
shock	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Acquisition	NN	O	O
of	NN	O	O
heat	NN	O	O
shock	NN	O	O
factor	NN	O	O
2	NN	O	O
(	NN	O	O
HSF2	NN	O	O
)	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
induced	NN	O	O
transcription	NN	O	O
of	NN	O	O
heat	NN	O	B-DNA
shock	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
hemin-treated	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
undergoing	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
revealed	NN	O	O
that	NN	O	O
HSF2	NN	O	B-protein
consists	NN	O	O
of	NN	O	O
two	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
isoforms	NN	O	O
,	NN	O	O
HSF2-alpha	NN	O	B-protein
and	NN	O	O
HSF2-beta	NN	O	B-protein
,	NN	O	O
whose	NN	O	O
relative	NN	O	O
abundance	NN	O	O
is	NN	O	O
developmentally	NN	O	O
regulated	NN	O	O
and	NN	O	O
varies	NN	O	O
between	NN	O	O
different	NN	O	O
tissues	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
whether	NN	O	O
the	NN	O	O
molar	NN	O	O
ratio	NN	O	O
of	NN	O	O
HSF2-alpha	NN	O	B-protein
and	NN	O	O
HSF2-beta	NN	O	B-protein
isoforms	NN	O	O
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
HSF2	NN	O	B-protein
and	NN	O	O
whether	NN	O	O
the	NN	O	O
HSF2	NN	O	B-protein
isoforms	NN	O	O
play	NN	O	O
functionally	NN	O	O
distinct	NN	O	O
roles	NN	O	O
during	NN	O	O
the	NN	O	O
hemin-mediated	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
cell	NN	O	O
clones	NN	O	O
expressing	NN	O	O
different	NN	O	O
levels	NN	O	O
of	NN	O	O
HSF2-alpha	NN	O	B-protein
and	NN	O	O
HSF2-beta	NN	O	B-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
parental	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
HSF2-alpha	NN	O	B-protein
isoform	NN	O	I-protein
is	NN	O	O
predominantly	NN	O	O
expressed	NN	O	O
and	NN	O	O
HSF2	NN	O	B-protein
can	NN	O	O
be	NN	O	O
activated	NN	O	O
upon	NN	O	O
hemin	NN	O	O
treatment	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
when	NN	O	O
HSF2-beta	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
levels	NN	O	O
exceeding	NN	O	O
those	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
HSF2-alpha	NN	O	I-protein
,	NN	O	O
the	NN	O	O
hemin-induced	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
transcription	NN	O	O
of	NN	O	O
heat	NN	O	B-DNA
shock	NN	O	I-DNA
genes	NN	O	I-DNA
are	NN	O	O
repressed	NN	O	O
,	NN	O	O
whereas	NN	O	O
overexpression	NN	O	O
of	NN	O	O
HSF2-alpha	NN	O	B-protein
results	NN	O	O
in	NN	O	O
an	NN	O	O
enhanced	NN	O	O
hemin	NN	O	B-protein
response	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
hemin	NN	O	B-protein
-induced	NN	O	O
accumulation	NN	O	O
of	NN	O	O
globin	NN	O	B-protein
,	NN	O	O
known	NN	O	O
as	NN	O	O
a	NN	O	O
marker	NN	O	O
of	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
,	NN	O	O
is	NN	O	O
decreased	NN	O	O
in	NN	O	O
cells	NN	O	O
overexpressing	NN	O	O
HSF2-beta	NN	O	B-protein
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
HSF2-beta	NN	O	B-protein
acts	NN	O	O
as	NN	O	O
a	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
HSF2	NN	O	B-protein
activity	NN	O	O
during	NN	O	O
hemin-mediated	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Sequential	NN	O	O
development	NN	O	O
of	NN	O	O
structural	NN	O	O
and	NN	O	O
functional	NN	O	O
alterations	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
tumor-bearing	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
TCR	NN	O	B-protein
alpha	NN	O	I-protein
beta	NN	O	I-protein
or	NN	O	I-protein
-gamma	NN	O	I-protein
delta	NN	O	I-protein
chains	NN	O	I-protein
bind	NN	O	O
the	NN	O	O
peptide	NN	O	O
ligand	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
associated	NN	O	O
CD3	NN	O	B-protein
delta	NN	O	I-protein
epsilon	NN	O	I-protein
gamma	NN	O	I-protein
and	NN	O	O
TCR	NN	O	B-protein
zeta	NN	O	I-protein
subunits	NN	O	I-protein
couple	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
to	NN	O	O
intracellular	NN	O	O
signal	NN	O	O
transduction	NN	O	O
components	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
several	NN	O	O
groups	NN	O	O
have	NN	O	O
described	NN	O	O
marked	NN	O	O
alterations	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
elements	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
cancer	NN	O	O
patients	NN	O	O
or	NN	O	O
in	NN	O	O
mice	NN	O	O
bearing	NN	O	O
tumor	NN	O	O
for	NN	O	O
a	NN	O	O
few	NN	O	O
weeks	NN	O	O
(	NN	O	O
>	NN	O	O
26	NN	O	O
days	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
sequence	NN	O	O
in	NN	O	O
which	NN	O	O
these	NN	O	O
alterations	NN	O	O
develop	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
explore	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
alterations	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
molecules	NN	O	O
(	NN	O	O
TCR	NN	O	B-protein
zeta	NN	O	I-protein
chain	NN	O	I-protein
,	NN	O	O
NF	NN	O	B-protein
kappaB	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
and	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
p56	NN	O	I-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	O
in	NN	O	O
mice	NN	O	O
bearing	NN	O	O
MC38	NN	O	O
colon	NN	O	O
adenocarcinoma	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
alterations	NN	O	O
in	NN	O	O
NF	NN	O	B-protein
kappaB	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
specifically	NN	O	O
the	NN	O	O
failure	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
translocation	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
occur	NN	O	O
earlier	NN	O	O
and	NN	O	O
more	NN	O	O
frequently	NN	O	O
than	NN	O	O
the	NN	O	O
decrease	NN	O	O
in	NN	O	O
zeta-chain	NN	O	B-protein
.	NN	O	O

These	NN	O	O
defects	NN	O	O
are	NN	O	O
paralleled	NN	O	O
by	NN	O	O
an	NN	O	O
impaired	NN	O	O
ability	NN	O	O
to	NN	O	O
produce	NN	O	O
Th1	NN	O	B-protein
cytokines	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
.	NN	O	O

These	NN	O	O
initial	NN	O	O
changes	NN	O	O
are	NN	O	O
followed	NN	O	O
by	NN	O	O
the	NN	O	O
eventual	NN	O	O
loss	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
zeta	NN	O	I-protein
chain	NN	O	I-protein
and	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
a	NN	O	O
marked	NN	O	O
decrease	NN	O	O
in	NN	O	O
cytotoxic	NN	O	O
function	NN	O	O
.	NN	O	O

An	NN	O	O
increased	NN	O	O
rate	NN	O	O
of	NN	O	O
lysosomal	NN	O	O
degradation	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
zeta-chain	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

HLA-DMA	NN	O	O
and	NN	O	O
HLA-DMB	NN	O	O
gene	NN	O	O
expression	NN	O	O
functions	NN	O	O
through	NN	O	O
the	NN	O	O
conserved	NN	O	O
S-X-Y	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
homologous	NN	O	I-protein
proteins	NN	O	I-protein
HLA-DMA	NN	O	B-protein
and	NN	O	O
HLA-DMB	NN	O	B-protein
function	NN	O	O
in	NN	O	O
the	NN	O	O
loading	NN	O	O
of	NN	O	O
peptides	NN	O	O
onto	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

Like	NN	O	O
the	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
HLA-DM	NN	O	B-DNA
genes	NN	O	I-DNA
contain	NN	O	O
upstream	NN	O	O
regulatory	NN	O	O
sequences	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
S-X-Y	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
additional	NN	O	O
putative	NN	O	O
regulatory	NN	O	O
sites	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
DM	NN	O	B-DNA
genes	NN	O	I-DNA
are	NN	O	O
regulated	NN	O	O
in	NN	O	O
a	NN	O	O
similar	NN	O	O
manner	NN	O	O
as	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
analyses	NN	O	O
was	NN	O	O
performed	NN	O	O
.	NN	O	O

Deletion	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
expression	NN	O	O
from	NN	O	O
the	NN	O	O
DM	NN	O	B-DNA
promoters	NN	O	I-DNA
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
conserved	NN	O	O
S-X-Y	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
class	NN	O	B-protein
II-specific	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
RFX	NN	O	B-protein
and	NN	O	O
CIITA	NN	O	B-protein
are	NN	O	O
also	NN	O	O
required	NN	O	O
for	NN	O	O
expression	NN	O	O
,	NN	O	O
as	NN	O	O
cell	NN	O	O
lines	NN	O	O
deficient	NN	O	O
in	NN	O	O
these	NN	O	O
factors	NN	O	O
failed	NN	O	O
to	NN	O	O
allow	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
DM	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprint	NN	O	O
analysis	NN	O	O
showed	NN	O	O
the	NN	O	O
putative	NN	O	O
X	NN	O	B-DNA
and	NN	O	I-DNA
Y	NN	O	I-DNA
boxes	NN	O	I-DNA
to	NN	O	O
be	NN	O	O
occupied	NN	O	O
by	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
wild-type	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
RFX-deficient	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
astrocytes	NN	O	B-cell_type
,	NN	O	O
IFN-gamma	NN	O	B-protein
treatment	NN	O	O
induced	NN	O	O
increased	NN	O	O
occupancy	NN	O	O
of	NN	O	O
these	NN	O	O
sites	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
putative	NN	O	O
regulatory	NN	O	O
sites	NN	O	O
was	NN	O	O
occupied	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
they	NN	O	O
may	NN	O	O
not	NN	O	O
be	NN	O	O
functional	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
gel	NN	O	O
shift	NN	O	O
analysis	NN	O	O
showed	NN	O	O
synergistic	NN	O	O
complex	NN	O	O
formation	NN	O	O
between	NN	O	O
proteins	NN	O	O
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
putative	NN	O	O
X	NN	O	B-DNA
boxes	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
DM	NN	O	O
genes	NN	O	O
,	NN	O	O
as	NN	O	O
is	NN	O	O
found	NN	O	O
for	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
DM	NN	O	B-DNA
genes	NN	O	I-DNA
share	NN	O	O
a	NN	O	O
common	NN	O	O
mechanism	NN	O	O
of	NN	O	O
regulation	NN	O	O
with	NN	O	O
the	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Effect	NN	O	O
of	NN	O	O
adenovirus	NN	O	O
2	NN	O	O
on	NN	O	O
cellular	NN	O	O
gene	NN	O	O
activation	NN	O	O
in	NN	O	O
blood-derived	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
adenovirus	NN	O	O
2	NN	O	O
(	NN	O	O
Ad2	NN	O	O
)	NN	O	O
infection	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
with	NN	O	O
regard	NN	O	O
to	NN	O	O
expression	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
.	NN	O	O

In	NN	O	O
monocytes	NN	O	O
,	NN	O	O
the	NN	O	O
virus	NN	O	O
was	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
surface	NN	O	O
without	NN	O	O
being	NN	O	O
internalized	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
Ad2	NN	O	O
was	NN	O	O
internalized	NN	O	O
by	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

No	NN	O	O
virus	NN	O	O
replication	NN	O	O
and	NN	O	O
no	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
Ad2	NN	O	B-DNA
early	NN	O	I-DNA
genes	NN	O	I-DNA
was	NN	O	O
observed	NN	O	O
in	NN	O	O
either	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

Ad2	NN	O	O
infection	NN	O	O
induced	NN	O	O
transient	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
levels	NN	O	O
for	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
in	NN	O	O
both	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
although	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
was	NN	O	O
slightly	NN	O	O
different	NN	O	O
.	NN	O	O

The	NN	O	O
production	NN	O	O
of	NN	O	O
both	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
measured	NN	O	O
by	NN	O	O
ELISA	NN	O	O
tests	NN	O	O
,	NN	O	O
was	NN	O	O
enhanced	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
a	NN	O	O
slight	NN	O	O
enhancement	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
was	NN	O	O
seen	NN	O	O
,	NN	O	O
whereas	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
was	NN	O	O
not	NN	O	O
detected	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
might	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
Ad2	NN	O	O
virus	NN	O	O
infection	NN	O	O
in	NN	O	O
nonpermissive	NN	O	O
cells	NN	O	O
where	NN	O	O
no	NN	O	O
viral	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
could	NN	O	O
be	NN	O	O
detected	NN	O	O
.	NN	O	O

-DOCSTART-	O

Structure	NN	O	O
and	NN	O	O
function	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
myeloid	NN	O	I-DNA
cell	NN	O	I-DNA
nuclear	NN	O	I-DNA
differentiation	NN	O	I-DNA
antigen	NN	O	I-DNA
promoter	NN	O	I-DNA
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
not	NN	O	O
of	NN	O	O
c-Myb	NN	O	B-protein
or	NN	O	O
PU.1	NN	O	B-protein
in	NN	O	O
myelomonocytic	NN	O	O
lineage-specific	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-protein
myeloid	NN	O	I-protein
nuclear	NN	O	I-protein
differentiation	NN	O	I-protein
antigen	NN	O	I-protein
(	NN	O	O
MNDA	NN	O	B-protein
)	NN	O	O
is	NN	O	O
expressed	NN	O	O
specifically	NN	O	O
in	NN	O	O
maturing	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
myelomonocytic	NN	O	O
lineage	NN	O	O
and	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

Epitope	NN	O	O
enhancement	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
confirm	NN	O	O
the	NN	O	O
strict	NN	O	O
lineage-	NN	O	O
and	NN	O	O
stage-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
MNDA	NN	O	B-protein
in	NN	O	O
bone	NN	O	O
marrow	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
other	NN	O	O
paraffin-embedded	NN	O	O
fixed	NN	O	O
tissues	NN	O	O
.	NN	O	O

A	NN	O	O
1-kb	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
gene	NN	O	O
that	NN	O	O
includes	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
sequence	NN	O	I-DNA
was	NN	O	O
reported	NN	O	O
earlier	NN	O	O
to	NN	O	O
contain	NN	O	O
functional	NN	O	O
promoter	NN	O	O
activity	NN	O	O
and	NN	O	O
was	NN	O	O
specifically	NN	O	O
demethylated	NN	O	O
in	NN	O	O
expressing	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
null	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Further	NN	O	O
analysis	NN	O	O
has	NN	O	O
revealed	NN	O	O
that	NN	O	O
this	NN	O	O
1-kb	NN	O	B-DNA
fragment	NN	O	I-DNA
promotes	NN	O	O
higher	NN	O	O
reporter	NN	O	O
gene	NN	O	O
activity	NN	O	O
in	NN	O	O
MNDA	NN	O	B-protein
-expressing	NN	O	B-cell_type
cells	NN	O	I-cell_type
than	NN	O	O
non-expressing	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
indicating	NN	O	O
cell-specific	NN	O	O
differences	NN	O	O
in	NN	O	O
transactivation	NN	O	O
.	NN	O	O

This	NN	O	O
sequence	NN	O	O
contains	NN	O	O
consensus	NN	O	B-DNA
elements	NN	O	I-DNA
consistent	NN	O	O
with	NN	O	O
myeloid-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
PU.1	NN	O	B-DNA
consensus	NN	O	I-DNA
site	NN	O	I-DNA
near	NN	O	O
the	NN	O	O
major	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
cluster	NN	O	O
of	NN	O	O
c-Myb	NN	O	B-DNA
sites	NN	O	I-DNA
located	NN	O	O
several	NN	O	O
hundred	NN	O	O
bases	NN	O	O
upstream	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
analysis	NN	O	O
of	NN	O	O
deletion	NN	O	B-DNA
mutants	NN	O	I-DNA
localized	NN	O	O
nearly	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
to	NN	O	O
a	NN	O	O
short	NN	O	O
region	NN	O	O
(	NN	O	O
-73	NN	O	B-DNA
to	NN	O	I-DNA
-16	NN	O	I-DNA
)	NN	O	O
that	NN	O	O
did	NN	O	O
not	NN	O	O
include	NN	O	O
the	NN	O	O
cluster	NN	O	O
of	NN	O	O
c-Myb	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
4-bp	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
Sp1	NN	O	I-DNA
consensus	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
GC	NN	O	B-DNA
box	NN	O	I-DNA
)	NN	O	O
(	NN	O	O
-20	NN	O	O
)	NN	O	O
reduced	NN	O	O
overall	NN	O	O
promoter	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
1-kb	NN	O	B-DNA
fragment	NN	O	I-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
PU.1	NN	O	B-DNA
site	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
significantly	NN	O	O
affect	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Only	NN	O	O
a	NN	O	O
small	NN	O	O
region	NN	O	O
(	NN	O	O
-35	NN	O	O
to	NN	O	O
+22	NN	O	O
)	NN	O	O
including	NN	O	O
the	NN	O	O
Sp1	NN	O	B-DNA
element	NN	O	I-DNA
and	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
PU.1	NN	O	B-DNA
site	NN	O	I-DNA
was	NN	O	O
footprinted	NN	O	O
.	NN	O	O

The	NN	O	O
4-bp	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
core	NN	O	O
Sp1	NN	O	B-DNA
consensus	NN	O	I-DNA
element	NN	O	I-DNA
abolished	NN	O	O
footprinting	NN	O	O
at	NN	O	O
the	NN	O	O
site	NN	O	O
and	NN	O	O
an	NN	O	O
antibody	NN	O	O
super-shift	NN	O	O
reaction	NN	O	O
showed	NN	O	O
that	NN	O	O
Sp1	NN	O	B-protein
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
factors	NN	O	O
binding	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
also	NN	O	O
co-localizes	NN	O	O
with	NN	O	O
a	NN	O	O
DNase	NN	O	B-DNA
I	NN	O	I-DNA
hypersensitive	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
DNA	NN	O	O
methylation	NN	O	O
,	NN	O	O
chromatin	NN	O	B-DNA
structure	NN	O	O
,	NN	O	O
and	NN	O	O
transactivation	NN	O	O
at	NN	O	O
an	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
highly	NN	O	O
restricted	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
myelomonocytic	NN	O	B-DNA
lineage	NN	O	I-DNA
specific	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Critical	NN	O	O
cytoplasmic	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
human	NN	O	B-protein
interleukin-9	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
in	NN	O	O
interleukin	NN	O	B-protein
-9-mediated	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

Interleukin-9	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
IL-9R	NN	O	I-protein
)	NN	O	I-protein
complex	NN	O	I-protein
consists	NN	O	O
of	NN	O	O
a	NN	O	O
ligand-specific	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
and	NN	O	O
IL-2R	NN	O	B-protein
gamma	NN	O	I-protein
chain	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
two	NN	O	O
regions	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
human	NN	O	B-protein
IL-9Ralpha	NN	O	I-protein
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
IL-9	NN	O	B-protein
-mediated	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

A	NN	O	O
membrane-proximal	NN	O	B-protein
region	NN	O	I-protein
that	NN	O	O
contains	NN	O	O
the	NN	O	O
BOX1	NN	O	B-protein
consensus	NN	O	I-protein
sequence	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
IL-9	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
kinases	NN	O	I-protein
(	NN	O	O
JAKs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
or	NN	O	O
internal	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
BOX1	NN	O	B-protein
motif	NN	O	I-protein
abrogated	NN	O	O
IL-9	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
substitution	NN	O	O
of	NN	O	O
the	NN	O	O
Pro-X-Pro	NN	O	B-protein
in	NN	O	O
the	NN	O	O
BOX1	NN	O	B-protein
motif	NN	O	I-protein
with	NN	O	O
Ala-X-Ala	NN	O	B-protein
failed	NN	O	O
to	NN	O	O
abolish	NN	O	O
IL-9	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
but	NN	O	O
decreased	NN	O	O
IL-9	NN	O	B-protein
-mediated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
JAK	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
insulin	NN	O	B-protein
receptor	NN	O	I-protein
substrate-2	NN	O	I-protein
,	NN	O	O
and	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
STAT3	NN	O	B-protein
)	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
c-myc	NN	O	B-DNA
and	NN	O	O
junB	NN	O	B-DNA
.	NN	O	O

Another	NN	O	O
important	NN	O	O
region	NN	O	O
is	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
BOX1	NN	O	B-protein
motif	NN	O	I-protein
and	NN	O	O
contains	NN	O	O
a	NN	O	O
STAT3	NN	O	B-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
YLPQ	NN	O	B-DNA
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
significantly	NN	O	O
impaired	NN	O	O
IL-9	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
JAK	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
insulin	NN	O	B-protein
receptor	NN	O	I-protein
substrate-2	NN	O	I-protein
,	NN	O	O
and	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
expression	NN	O	O
of	NN	O	O
early	NN	O	B-DNA
response	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
point	NN	O	O
mutation	NN	O	O
changing	NN	O	O
YLPQ	NN	O	B-DNA
into	NN	O	O
YLPA	NN	O	B-DNA
greatly	NN	O	O
reduced	NN	O	O
IL-9	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
and	NN	O	O
expression	NN	O	O
of	NN	O	O
c-myc	NN	O	B-DNA
but	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
cooperation	NN	O	O
or	NN	O	O
cross-talk	NN	O	O
of	NN	O	O
signaling	NN	O	O
molecules	NN	O	O
associated	NN	O	O
with	NN	O	O
different	NN	O	O
domains	NN	O	O
of	NN	O	O
IL-9Ralpha	NN	O	B-protein
other	NN	O	O
than	NN	O	O
STAT3	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
IL-9	NN	O	B-protein
-mediated	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

-DOCSTART-	O

Bcl-2	NN	O	B-protein
protein	NN	O	I-protein
inhibits	NN	O	O
bufalin-induced	NN	O	O
apoptosis	NN	O	O
through	NN	O	O
inhibition	NN	O	O
of	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
a	NN	O	O
previous	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
bufalin	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
an	NN	O	O
active	NN	O	O
principle	NN	O	O
of	NN	O	O
Chinese	NN	O	O
medicine	NN	O	O
,	NN	O	O
chan'su	NN	O	O
,	NN	O	O
caused	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
anomalous	NN	O	O
activation	NN	O	O
of	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
via	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
of	NN	O	O
Ras	NN	O	B-protein
,	NN	O	O
Raf-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
MAPK	NN	O	B-protein
kinase-1	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
overexpression	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
on	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
that	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
bufalin	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
bufalin	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
Bcl-2	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

No	NN	O	O
significant	NN	O	O
difference	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
kinase-1	NN	O	I-protein
that	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
bufalin	NN	O	O
in	NN	O	O
wild-type	NN	O	O
or	NN	O	O
Bcl-2-overexpressed	NN	O	B-cell_type
U937	NN	O	I-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
by	NN	O	O
bufalin	NN	O	O
was	NN	O	O
significantly	NN	O	O
attenuated	NN	O	O
in	NN	O	O
the	NN	O	O
cells	NN	O	O
overexpressing	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

Bufalin	NN	O	O
treatment	NN	O	O
activated	NN	O	B-protein
activator	NN	O	I-protein
protein-1	NN	O	I-protein
transcriptional	NN	O	O
activity	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
this	NN	O	O
activation	NN	O	O
was	NN	O	O
decreased	NN	O	O
to	NN	O	O
40	NN	O	O
%	NN	O	O
in	NN	O	O
bcl-2	NN	O	B-protein
-overexpressed	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
Bcl-2	NN	O	B-protein
acts	NN	O	O
downstream	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
kinase-1	NN	O	I-protein
but	NN	O	O
upstream	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
of	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
process	NN	O	O
induced	NN	O	O
by	NN	O	O
bufalin	NN	O	O
,	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
may	NN	O	O
be	NN	O	O
down-regulated	NN	O	O
through	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Pancreatic	NN	O	O
islet	NN	O	O
expression	NN	O	O
studies	NN	O	O
and	NN	O	O
polymorphic	NN	O	O
DNA	NN	O	O
markers	NN	O	O
in	NN	O	O
the	NN	O	O
genes	NN	O	O
encoding	NN	O	O
hepatocyte	NN	O	B-protein
nuclear	NN	O	I-protein
factor-3alpha	NN	O	I-protein
,	NN	O	I-protein
-3beta	NN	O	I-protein
,	NN	O	I-protein
-3gamma	NN	O	I-protein
,	NN	O	I-protein
-4gamma	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
-6	NN	O	I-protein
.	NN	O	O

The	NN	O	O
genes	NN	O	O
encoding	NN	O	O
the	NN	O	O
functionally	NN	O	O
related	NN	O	O
hepatocyte	NN	O	B-protein
nuclear	NN	O	I-protein
factors	NN	O	I-protein
HNF-1alpha	NN	O	B-protein
and	NN	O	O
HNF-4alpha	NN	O	B-protein
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
normal	NN	O	O
pancreatic	NN	O	O
beta-cell	NN	O	O
function	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
these	NN	O	O
liver-enriched	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
result	NN	O	O
in	NN	O	O
two	NN	O	O
forms	NN	O	O
of	NN	O	O
early-onset	NN	O	O
type	NN	O	O
2	NN	O	O
diabetes	NN	O	O
(	NN	O	O
maturity-onset	NN	O	O
diabetes	NN	O	O
of	NN	O	O
the	NN	O	O
young	NN	O	O
[	NN	O	O
MODY	NN	O	O
]	NN	O	O
)	NN	O	O
,	NN	O	O
MODY3	NN	O	O
and	NN	O	O
MODY1	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
characterized	NN	O	O
by	NN	O	O
impaired	NN	O	O
glucose-stimulated	NN	O	O
insulin	NN	O	O
secretion	NN	O	O
,	NN	O	O
early	NN	O	O
disease	NN	O	O
onset	NN	O	O
,	NN	O	O
and	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
inheritance	NN	O	O
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	O
hierarchy	NN	O	O
of	NN	O	O
HNFs	NN	O	O
suggests	NN	O	O
that	NN	O	O
other	NN	O	O
proteins	NN	O	O
of	NN	O	O
the	NN	O	O
regulatory	NN	O	O
cascade	NN	O	O
might	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
other	NN	O	O
forms	NN	O	O
of	NN	O	O
MODY	NN	O	O
and/or	NN	O	O
late-onset	NN	O	O
type	NN	O	O
2	NN	O	O
diabetes	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
HNF-3alpha	NN	O	B-protein
,	NN	O	I-protein
-3beta	NN	O	I-protein
,	NN	O	I-protein
-3gamma	NN	O	I-protein
,	NN	O	I-protein
-4gamma	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
-6	NN	O	I-protein
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
pancreatic	NN	O	B-cell_type
beta-cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
identification	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
simple	NN	O	B-DNA
tandem	NN	O	I-DNA
repeat	NN	O	I-DNA
DNA	NN	O	I-DNA
polymorphisms	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
genes	NN	O	O
encoding	NN	O	O
HNF-3alpha	NN	O	B-protein
,	NN	O	O
-3beta	NN	O	B-protein
,	NN	O	O
-3gamma	NN	O	B-protein
,	NN	O	O
-4gamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
-6	NN	O	B-protein
and	NN	O	O
the	NN	O	O
mapping	NN	O	O
of	NN	O	O
HNF-6	NN	O	B-protein
to	NN	O	O
chromosome	NN	O	B-DNA
bands	NN	O	I-DNA
15q21.1-21.2	NN	O	B-DNA
by	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

These	NN	O	O
markers	NN	O	O
will	NN	O	O
be	NN	O	O
useful	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
genetic	NN	O	O
variation	NN	O	O
in	NN	O	O
these	NN	O	O
genes	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
type	NN	O	O
2	NN	O	O
diabetes	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B/Rel	NN	O	I-protein
nuclear	NN	O	O
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
anti-HLA	NN	O	B-protein
class	NN	O	I-protein
I	NN	O	I-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

Monoclonal	NN	O	O
antibodies	NN	O	O
against	NN	O	O
either	NN	O	O
monomorphic	NN	O	O
or	NN	O	O
polymorphic	NN	O	O
determinants	NN	O	O
of	NN	O	O
class	NN	O	B-protein
I	NN	O	I-protein
antigen	NN	O	I-protein
induced	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
and	NN	O	O
highly	NN	O	O
purified	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
the	NN	O	O
nuclear	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

These	NN	O	O
included	NN	O	O
both	NN	O	O
p50/p50	NN	O	B-protein
and	NN	O	O
p50/p65	NN	O	B-protein
dimers	NN	O	I-protein
,	NN	O	O
recognized	NN	O	O
by	NN	O	O
specific	NN	O	B-protein
antibodies	NN	O	I-protein
in	NN	O	O
EMSA	NN	O	O
.	NN	O	O

The	NN	O	O
induced	NN	O	O
complexes	NN	O	B-protein
were	NN	O	O
detectable	NN	O	O
in	NN	O	O
extracts	NN	O	O
of	NN	O	O
cells	NN	O	O
incubated	NN	O	O
with	NN	O	O
anti-class	NN	O	B-protein
I	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
mAb	NN	O	B-protein
)	NN	O	O
for	NN	O	O
1.5	NN	O	O
h	NN	O	O
;	NN	O	O
the	NN	O	O
induction	NN	O	O
was	NN	O	O
maximal	NN	O	O
at	NN	O	O
5	NN	O	O
h	NN	O	O
,	NN	O	O
persistent	NN	O	O
at	NN	O	O
16	NN	O	O
h	NN	O	O
and	NN	O	O
no	NN	O	O
longer	NN	O	O
observed	NN	O	O
at	NN	O	O
40	NN	O	O
h	NN	O	O
.	NN	O	O

The	NN	O	O
mAb	NN	O	B-protein
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
nuclear	NN	O	O
activity	NN	O	O
in	NN	O	O
cells	NN	O	O
incubated	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
3	NN	O	O
,	NN	O	O
4-dichloroisocoumarin	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
degradation	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
class	NN	O	O
I	NN	O	O
triggering	NN	O	O
can	NN	O	O
induce	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
nuclear	NN	O	O
activity	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
thereby	NN	O	O
modulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
regulated	NN	O	O
by	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Anti-Ehrlichia	NN	O	B-protein
chaffeensis	NN	O	I-protein
antibody	NN	O	I-protein
complexed	NN	O	O
with	NN	O	O
E.	NN	O	O
chaffeensis	NN	O	O
induces	NN	O	O
potent	NN	O	O
proinflammatory	NN	O	B-RNA
cytokine	NN	O	I-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
through	NN	O	O
sustained	NN	O	O
reduction	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
and	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Ehrlichia	NN	O	O
chaffeensis	NN	O	O
is	NN	O	O
an	NN	O	O
obligatory	NN	O	O
intracellular	NN	O	O
bacterium	NN	O	O
that	NN	O	O
infects	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
is	NN	O	O
the	NN	O	O
etiologic	NN	O	O
agent	NN	O	O
of	NN	O	O
human	NN	O	O
ehrlichiosis	NN	O	O
in	NN	O	O
the	NN	O	O
United	NN	O	O
States	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
exposure	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
to	NN	O	O
E.	NN	O	O
chaffeensis	NN	O	O
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
interleukin-1beta	NN	O	B-protein
(	NN	O	O
IL-1beta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-10	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
tumor	NN	O	B-RNA
necrosis	NN	O	I-RNA
factor	NN	O	I-RNA
alpha	NN	O	I-RNA
(	NN	O	I-RNA
TNF-alpha	NN	O	I-RNA
)	NN	O	I-RNA
and	NN	O	I-RNA
IL-6	NN	O	I-RNA
mRNAs	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
anti-E.	NN	O	B-protein
chaffeensis	NN	O	I-protein
antibody	NN	O	I-protein
complexed	NN	O	O
with	NN	O	O
E.	NN	O	O
chaffeensis	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
major	NN	O	B-protein
proinflammatory	NN	O	I-protein
cytokines	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
was	NN	O	O
examined	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
was	NN	O	O
treated	NN	O	O
with	NN	O	O
E.	NN	O	O
chaffeensis	NN	O	O
which	NN	O	O
had	NN	O	O
been	NN	O	O
preincubated	NN	O	O
with	NN	O	O
human	NN	O	O
anti-E.	NN	O	O
chaffeensis	NN	O	O
serum	NN	O	O
for	NN	O	O
2	NN	O	O
h	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
cytokine	NN	O	B-RNA
mRNAs	NN	O	I-RNA
were	NN	O	O
evaluated	NN	O	O
by	NN	O	O
competitive	NN	O	O
reverse	NN	O	O
transcription-PCR	NN	O	O
.	NN	O	O

Anti-E.	NN	O	B-protein
chaffeensis	NN	O	I-protein
antibody	NN	O	I-protein
complexed	NN	O	O
with	NN	O	O
E.	NN	O	O
chaffeensis	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-RNA
and	NN	O	I-RNA
IL-6	NN	O	I-RNA
mRNAs	NN	O	I-RNA
was	NN	O	O
also	NN	O	O
induced	NN	O	O
.	NN	O	O

The	NN	O	O
levels	NN	O	O
of	NN	O	O
secreted	NN	O	O
IL-1beta	NN	O	B-protein
,	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-6	NN	O	B-protein
during	NN	O	O
24	NN	O	O
h	NN	O	O
of	NN	O	O
stimulation	NN	O	O
were	NN	O	O
comparable	NN	O	O
to	NN	O	O
those	NN	O	O
induced	NN	O	O
by	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
lipopolysaccharide	NN	O	O
at	NN	O	O
1	NN	O	O
microg/ml	NN	O	O
.	NN	O	O

Fab	NN	O	O
fragment	NN	O	O
of	NN	O	O
anti-E.	NN	O	B-protein
chaffeensis	NN	O	I-protein
immunoglobulin	NN	O	I-protein
G	NN	O	I-protein
complexed	NN	O	O
with	NN	O	O
E.	NN	O	O
chaffeensis	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
any	NN	O	O
of	NN	O	O
these	NN	O	O
three	NN	O	O
cytokines	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
ehrlichial	NN	O	O
binding	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
IL-1beta	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	B-protein
complex	NN	O	I-protein
to	NN	O	O
the	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
receptor	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
enhanced	NN	O	O
IL-1beta	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
prolonged	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
and	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
were	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
exposed	NN	O	O
to	NN	O	O
anti-E.	NN	O	O
chaffeensis	NN	O	O
serum	NN	O	O
and	NN	O	O
E.	NN	O	O
chaffeensis	NN	O	O
.	NN	O	O

This	NN	O	O
result	NN	O	O
implies	NN	O	O
that	NN	O	O
development	NN	O	O
of	NN	O	O
anti-E.	NN	O	B-protein
chaffeensis	NN	O	I-protein
antibody	NN	O	I-protein
in	NN	O	O
patients	NN	O	O
can	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
major	NN	O	B-protein
proinflammatory	NN	O	I-protein
cytokines	NN	O	I-protein
,	NN	O	O
which	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
ehrlichiosis	NN	O	O
and	NN	O	O
immune	NN	O	O
responses	NN	O	O
to	NN	O	O
it	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
trans-activator	NN	O	I-protein
CIITA	NN	O	I-protein
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
TBP-associated	NN	O	B-protein
factor	NN	O	I-protein
TAFII32	NN	O	I-protein
.	NN	O	O

The	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
trans-	NN	O	I-protein
activator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
is	NN	O	O
the	NN	O	O
main	NN	O	O
transcriptional	NN	O	O
co-activator	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Its	NN	O	O
N-terminal	NN	O	B-protein
125	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
function	NN	O	O
as	NN	O	O
an	NN	O	O
independent	NN	O	B-protein
transcriptional	NN	O	I-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Analyses	NN	O	O
of	NN	O	O
the	NN	O	O
primary	NN	O	B-protein
amino	NN	O	I-protein
acid	NN	O	I-protein
sequence	NN	O	I-protein
of	NN	O	O
the	NN	O	O
activation	NN	O	B-protein
domain	NN	O	I-protein
predict	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
three	NN	O	O
alpha-helices	NN	O	B-protein
,	NN	O	O
each	NN	O	O
with	NN	O	O
a	NN	O	O
high	NN	O	O
proportion	NN	O	O
of	NN	O	O
acidic	NN	O	O
residues	NN	O	O
.	NN	O	O

Using	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
two	NN	O	O
of	NN	O	O
these	NN	O	O
predicted	NN	O	O
alpha-helices	NN	O	B-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
full	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
CIITA	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
a	NN	O	O
CIITA	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
which	NN	O	O
both	NN	O	O
functional	NN	O	O
alpha-helices	NN	O	B-protein
have	NN	O	O
been	NN	O	O
deleted	NN	O	O
displays	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
phenotype	NN	O	O
.	NN	O	O

This	NN	O	O
activation	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
CIITA	NN	O	B-protein
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
32	NN	O	B-protein
kDa	NN	O	I-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
general	NN	O	B-protein
transcription	NN	O	I-protein
complex	NN	O	I-protein
TFIID	NN	O	B-protein
,	NN	O	O
TAFII32	NN	O	B-protein
.	NN	O	O

Decreased	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
N-terminal	NN	O	B-protein
deletions	NN	O	I-protein
of	NN	O	O
CIITA	NN	O	B-protein
is	NN	O	O
correlated	NN	O	O
directly	NN	O	O
with	NN	O	O
their	NN	O	O
reduced	NN	O	O
binding	NN	O	O
to	NN	O	O
TAFII32	NN	O	B-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
interactions	NN	O	O
between	NN	O	O
TAFII32	NN	O	B-protein
and	NN	O	O
CIITA	NN	O	B-protein
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
interleukin-10	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
on	NN	O	O
B-cell	NN	O	B-cell_line
chronic	NN	O	I-cell_line
lymphocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

B-cell	NN	O	B-cell_line
chronic	NN	O	I-cell_line
lymphocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
(	NN	O	I-cell_line
B-CLL	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
accumulate	NN	O	O
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
the	NN	O	O
G0/G1	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
their	NN	O	O
malignant	NN	O	O
expansion	NN	O	O
is	NN	O	O
due	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
delay	NN	O	O
in	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
cellular	NN	O	O
or	NN	O	O
molecular	NN	O	B-protein
factors	NN	O	I-protein
responsible	NN	O	O
for	NN	O	O
a	NN	O	O
delay	NN	O	O
in	NN	O	O
B-CLL	NN	O	O
cell	NN	O	O
death	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

B-CLL	NN	O	O
cells	NN	O	O
do	NN	O	O
express	NN	O	O
receptors	NN	O	O
for	NN	O	O
interferon-alpha	NN	O	B-protein
(	NN	O	O
IFN-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
promote	NN	O	O
B-CLL	NN	O	O
survival	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
preventing	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
interleukin-10	NN	O	B-protein
(	NN	O	I-protein
IL-10	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
is	NN	O	O
another	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
IFN	NN	O	B-protein
receptor	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
but	NN	O	O
its	NN	O	O
ligand	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
B-CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
the	NN	O	O
current	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
undertook	NN	O	O
a	NN	O	O
biochemical	NN	O	O
analysis	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
on	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
B-CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
characterized	NN	O	O
the	NN	O	O
functional	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
its	NN	O	O
constitutively	NN	O	B-protein
expressed	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
B-CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
bind	NN	O	O
IL-10	NN	O	B-protein
with	NN	O	O
significant	NN	O	O
specificity	NN	O	O
and	NN	O	O
express	NN	O	O
between	NN	O	O
47	NN	O	O
and	NN	O	O
127	NN	O	O
IL-10	NN	O	B-protein
receptor	NN	O	I-protein
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
in	NN	O	O
the	NN	O	O
range	NN	O	O
of	NN	O	O
168	NN	O	O
to	NN	O	O
426	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-12	NN	O	O
)	NN	O	O
mol/L	NN	O	O
.	NN	O	O

Ligand	NN	O	O
binding	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-10	NN	O	B-protein
receptor	NN	O	I-protein
expressed	NN	O	O
on	NN	O	O
B-CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
results	NN	O	O
in	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
STAT1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
STAT3	NN	O	B-protein
proteins	NN	O	O
.	NN	O	O

This	NN	O	O
pattern	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
protein	NN	O	I-protein
phosphorylation	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
IL-10	NN	O	B-protein
receptor	NN	O	I-protein
activation	NN	O	O
on	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
similar	NN	O	O
to	NN	O	O
IFN-alpha	NN	O	B-protein
(	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
STAT3	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
(	NN	O	O
STAT1	NN	O	B-protein
)	NN	O	O
receptor	NN	O	O
activation	NN	O	O
in	NN	O	O
CLL	NN	O	B-cell_line
.	NN	O	O

Further	NN	O	O
,	NN	O	O
in	NN	O	O
consecutive	NN	O	O
samples	NN	O	O
of	NN	O	O
fresh	NN	O	O
blood	NN	O	O
obtained	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
B-CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
inhibited	NN	O	O
B-CLL	NN	O	B-cell_line
proliferation	NN	O	O
,	NN	O	O
enhanced	NN	O	O
B-CLL	NN	O	B-cell_line
differentiation	NN	O	O
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
,	NN	O	O
like	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
significantly	NN	O	O
reduce	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
B-CLL	NN	O	O
cell	NN	O	O
death	NN	O	O
caused	NN	O	O
by	NN	O	O
hydrocortisone-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
which	NN	O	O
signal	NN	O	O
through	NN	O	O
the	NN	O	O
interferon	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
receptors	NN	O	O
,	NN	O	O
have	NN	O	O
comparable	NN	O	O
functional	NN	O	O
effects	NN	O	O
on	NN	O	O
B-CLL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
mineralocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
hormone-binding	NN	O	I-protein
domain	NN	O	I-protein
in	NN	O	O
agonist	NN	O	O
and	NN	O	O
antagonist	NN	O	O
binding	NN	O	O
as	NN	O	O
revealed	NN	O	O
by	NN	O	O
a	NN	O	O
new	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
.	NN	O	O

To	NN	O	O
gain	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-protein
mineralocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
hMR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
developed	NN	O	O
a	NN	O	O
new	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
(	NN	O	O
mAb	NN	O	B-protein
)	NN	O	O
raised	NN	O	O
against	NN	O	O
the	NN	O	O
hormone-binding	NN	O	B-protein
domain	NN	O	I-protein
(	NN	O	O
HBD	NN	O	B-protein
)	NN	O	O
.	NN	O	O

For	NN	O	O
this	NN	O	O
purpose	NN	O	O
,	NN	O	O
mice	NN	O	O
were	NN	O	O
immunized	NN	O	O
with	NN	O	O
a	NN	O	O
fusion	NN	O	B-protein
protein	NN	O	I-protein
including	NN	O	O
the	NN	O	O
sequence	NN	O	O
Thr729-Lys984	NN	O	B-protein
of	NN	O	O
hMR	NN	O	B-protein
.	NN	O	O

After	NN	O	O
ELISA	NN	O	O
screening	NN	O	O
,	NN	O	O
mAb	NN	O	B-protein
18C7	NN	O	I-protein
was	NN	O	O
selected	NN	O	O
for	NN	O	O
its	NN	O	O
specificity	NN	O	O
towards	NN	O	O
the	NN	O	O
HBD	NN	O	B-protein
.	NN	O	O

This	NN	O	O
antibody	NN	O	O
recognized	NN	O	O
both	NN	O	O
the	NN	O	O
denatured	NN	O	O
and	NN	O	O
native	NN	O	B-protein
MR	NN	O	I-protein
forms	NN	O	I-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
hetero-oligomeric	NN	O	B-protein
MR	NN	O	I-protein
form	NN	O	I-protein
and	NN	O	O
the	NN	O	O
transformed	NN	O	B-protein
MR	NN	O	I-protein
state	NN	O	I-protein
.	NN	O	O

By	NN	O	O
using	NN	O	O
several	NN	O	O
HBD	NN	O	B-protein
subfragments	NN	O	I-protein
,	NN	O	O
the	NN	O	O
mAb	NN	O	B-protein
18C7	NN	O	I-protein
epitope	NN	O	I-protein
was	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HBD	NN	O	B-protein
from	NN	O	O
Thr729	NN	O	B-protein
to	NN	O	I-protein
Leu765	NN	O	I-protein
.	NN	O	O

We	NN	O	O
then	NN	O	O
studied	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
antibody	NN	O	O
on	NN	O	O
aldosterone	NN	O	O
and	NN	O	O
progesterone	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
hMR	NN	O	B-protein
.	NN	O	O

When	NN	O	O
18C7	NN	O	B-protein
was	NN	O	O
incubated	NN	O	O
with	NN	O	O
liganded	NN	O	B-protein
MR	NN	O	I-protein
,	NN	O	O
it	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
partly	NN	O	O
displace	NN	O	O
(	NN	O	O
20	NN	O	O
%	NN	O	O
)	NN	O	O
the	NN	O	O
hormone	NN	O	O
from	NN	O	O
its	NN	O	O
binding	NN	O	O
site	NN	O	O
.	NN	O	O

When	NN	O	O
18C7	NN	O	B-protein
was	NN	O	O
incubated	NN	O	O
with	NN	O	O
MR	NN	O	B-protein
before	NN	O	O
aldosterone	NN	O	O
or	NN	O	O
progesterone	NN	O	O
,	NN	O	O
the	NN	O	O
antibody	NN	O	O
inhibited	NN	O	O
75-80	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
18C7	NN	O	B-protein
on	NN	O	O
the	NN	O	O
binding	NN	O	O
was	NN	O	O
similar	NN	O	O
with	NN	O	O
both	NN	O	O
hormones	NN	O	O
.	NN	O	O

A	NN	O	O
sucrose	NN	O	O
gradient	NN	O	O
analysis	NN	O	O
indicated	NN	O	O
the	NN	O	O
simultaneous	NN	O	O
presence	NN	O	O
of	NN	O	O
two	NN	O	O
kinds	NN	O	O
of	NN	O	O
receptor	NN	O	O
complexes	NN	O	O
:	NN	O	O
the	NN	O	O
steroid-MR	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
the	NN	O	O
antibody-MR	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

After	NN	O	O
its	NN	O	O
associated	NN	O	O
proteins	NN	O	O
,	NN	O	O
especially	NN	O	O
the	NN	O	O
heat-shock	NN	O	B-protein
protein	NN	O	I-protein
hsp90	NN	O	I-protein
,	NN	O	O
had	NN	O	O
been	NN	O	O
cross-linked	NN	O	O
with	NN	O	O
the	NN	O	O
hMR	NN	O	B-protein
by	NN	O	O
dimethylpimelimidate	NN	O	O
,	NN	O	O
18C7	NN	O	B-protein
was	NN	O	O
still	NN	O	O
able	NN	O	O
to	NN	O	O
react	NN	O	O
with	NN	O	O
the	NN	O	O
receptor	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
epitope	NN	O	B-protein
recognized	NN	O	O
by	NN	O	O
18C7	NN	O	B-protein
was	NN	O	O
directly	NN	O	O
implicated	NN	O	O
in	NN	O	O
hormone	NN	O	O
binding	NN	O	O
.	NN	O	O

The	NN	O	O
lack	NN	O	O
of	NN	O	O
steroid	NN	O	O
binding	NN	O	O
of	NN	O	O
HBD	NN	O	B-protein
mutants	NN	O	O
with	NN	O	O
the	NN	O	O
Thr729-Leu765	NN	O	B-protein
sequence	NN	O	I-protein
deleted	NN	O	O
[	NN	O	O
Jalaguier	NN	O	O
,	NN	O	O
Mesnier	NN	O	O
,	NN	O	O
Leger	NN	O	O
and	NN	O	O
Auzou	NN	O	O
(	NN	O	O
1996	NN	O	O
)	NN	O	O
J.Steroid	NN	O	O
Biochem.Mol.Biol.57	NN	O	O
,	NN	O	O
43-50	NN	O	O
]	NN	O	O
supports	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
.	NN	O	O

Because	NN	O	O
of	NN	O	O
the	NN	O	O
similar	NN	O	O
behaviours	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
and	NN	O	O
progesterone	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
Thr729-Leu765	NN	O	I-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HBD	NN	O	B-protein
is	NN	O	O
similarly	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
both	NN	O	O
hormones	NN	O	O
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
cloning	NN	O	O
of	NN	O	O
SLAP-130	NN	O	B-protein
,	NN	O	O
an	NN	O	O
SLP-76-associated	NN	O	B-protein
substrate	NN	O	I-protein
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor-stimulated	NN	O	I-protein
protein	NN	O	I-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

Previous	NN	O	O
work	NN	O	O
has	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
SLP-76	NN	O	B-protein
,	NN	O	O
a	NN	O	O
Grb2-associated	NN	O	B-protein
tyrosine-phosphorylated	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
augments	NN	O	O
Interleukin-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
when	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
activity	NN	O	O
requires	NN	O	O
regions	NN	O	O
of	NN	O	O
SLP-76	NN	O	B-protein
that	NN	O	O
mediate	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
with	NN	O	O
other	NN	O	O
molecules	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
SLP-76	NN	O	B-protein
-associated	NN	O	O
proteins	NN	O	O
also	NN	O	O
function	NN	O	O
to	NN	O	O
regulate	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
molecular	NN	O	O
cloning	NN	O	O
of	NN	O	O
SLAP-130	NN	O	B-protein
,	NN	O	O
a	NN	O	O
SLP-76-associated	NN	O	B-protein
phosphoprotein	NN	O	I-protein
of	NN	O	O
130	NN	O	O
kDa	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
SLAP-130	NN	O	B-protein
is	NN	O	O
hematopoietic	NN	O	O
cell-specific	NN	O	O
and	NN	O	O
associates	NN	O	O
with	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
SLP-76	NN	O	B-protein
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
SLAP-130	NN	O	B-protein
is	NN	O	O
a	NN	O	O
substrate	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor-induced	NN	O	I-protein
protein	NN	O	I-protein
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
we	NN	O	O
find	NN	O	O
that	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
SLP-76	NN	O	B-protein
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
SLAP-130	NN	O	B-protein
diminishes	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
interferes	NN	O	O
with	NN	O	O
the	NN	O	O
augmentation	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
seen	NN	O	O
when	NN	O	O
SLP-76	NN	O	B-protein
is	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
SLP-76	NN	O	B-protein
recruits	NN	O	O
a	NN	O	O
negative	NN	O	O
regulator	NN	O	O
,	NN	O	O
SLAP-130	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
positive	NN	O	O
regulators	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
actions	NN	O	O
of	NN	O	O
prolactin	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

The	NN	O	O
immunoregulatory	NN	O	B-protein
properties	NN	O	O
of	NN	O	O
prolactin	NN	O	O
,	NN	O	O
a	NN	O	O
pituitary	NN	O	O
peptide	NN	O	O
hormone	NN	O	O
,	NN	O	O
have	NN	O	O
received	NN	O	O
renewed	NN	O	O
attention	NN	O	O
.	NN	O	O

The	NN	O	O
prolactin	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
hematopoietin/cytokine	NN	O	B-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
,	NN	O	O
is	NN	O	O
ubiquitously	NN	O	O
expressed	NN	O	O
by	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Certain	NN	O	O
subpopulations	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
synthesize	NN	O	O
and	NN	O	O
secrete	NN	O	O
biologically	NN	O	O
active	NN	O	O
prolactin	NN	O	O
,	NN	O	O
which	NN	O	O
suggests	NN	O	O
that	NN	O	O
prolactin	NN	O	O
can	NN	O	O
act	NN	O	O
as	NN	O	O
an	NN	O	O
autocrine	NN	O	O
and/or	NN	O	O
paracrine	NN	O	O
factor	NN	O	O
to	NN	O	O
modulate	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

This	NN	O	O
review	NN	O	O
focuses	NN	O	O
on	NN	O	O
the	NN	O	O
molecular	NN	O	O
actions	NN	O	O
of	NN	O	O
prolactin	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Emphasis	NN	O	O
is	NN	O	O
given	NN	O	O
to	NN	O	O
recent	NN	O	O
information	NN	O	O
about	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
prolactin	NN	O	O
receptor	NN	O	O
signal	NN	O	O
transduction	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
signaling	NN	O	B-protein
molecules	NN	O	I-protein
and	NN	O	O
prolactin-inducible	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
that	NN	O	O
participate	NN	O	O
in	NN	O	O
these	NN	O	O
responses	NN	O	O
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
the	NN	O	O
prolactin-inducible	NN	O	B-DNA
interferon	NN	O	I-DNA
regulatory	NN	O	I-DNA
factor-1	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
its	NN	O	O
roles	NN	O	O
in	NN	O	O
mediating	NN	O	O
diverse	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
during	NN	O	O
myelopoiesis	NN	O	O
.	NN	O	O

The	NN	O	O
coordinated	NN	O	O
production	NN	O	O
of	NN	O	O
all	NN	O	O
blood	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
a	NN	O	O
common	NN	O	B-cell_type
stem	NN	O	I-cell_type
cell	NN	O	I-cell_type
is	NN	O	O
a	NN	O	O
highly	NN	O	O
regulated	NN	O	O
process	NN	O	O
involving	NN	O	O
successive	NN	O	O
stages	NN	O	O
of	NN	O	O
commitment	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

From	NN	O	O
analyses	NN	O	O
of	NN	O	O
mice	NN	O	O
deficient	NN	O	O
in	NN	O	O
transcription	NN	O	B-DNA
factor	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
from	NN	O	O
the	NN	O	O
characterizations	NN	O	O
of	NN	O	O
chromosome	NN	O	B-DNA
breakpoints	NN	O	I-DNA
in	NN	O	O
human	NN	O	O
leukemias	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
become	NN	O	O
evident	NN	O	O
that	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
important	NN	O	O
regulators	NN	O	O
of	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

During	NN	O	O
myelopoiesis	NN	O	O
,	NN	O	O
which	NN	O	O
includes	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
granulocytic	NN	O	B-cell_type
and	NN	O	I-cell_type
monocytic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
,	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
from	NN	O	O
several	NN	O	O
families	NN	O	O
are	NN	O	O
active	NN	O	O
,	NN	O	O
including	NN	O	O
AML1/CBF	NN	O	B-DNA
beta	NN	O	I-DNA
,	NN	O	O
C/EBP	NN	O	B-DNA
,	NN	O	O
Ets	NN	O	B-DNA
,	NN	O	O
c-Myb	NN	O	B-DNA
,	NN	O	O
HOX	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
MZF-1	NN	O	B-DNA
.	NN	O	O

Few	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
are	NN	O	O
expressed	NN	O	O
exclusively	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
instead	NN	O	O
it	NN	O	O
appears	NN	O	O
that	NN	O	O
they	NN	O	O
cooperatively	NN	O	O
regulate	NN	O	O
transcription	NN	O	O
of	NN	O	O
myeloid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
we	NN	O	O
discuss	NN	O	O
recent	NN	O	O
advances	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
during	NN	O	O
myelopoiesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
transcription	NN	O	O
factor	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
phagocytic	NN	O	O
stimuli	NN	O	O
in	NN	O	O
human	NN	O	O
neutrophils	NN	O	O
.	NN	O	O

Phagocytosis	NN	O	O
represents	NN	O	O
an	NN	O	O
important	NN	O	O
physiological	NN	O	O
trigger	NN	O	O
for	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
genes	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
a	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
primarily	NN	O	O
consisting	NN	O	O
of	NN	O	O
the	NN	O	O
classical	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
heterodimer	NN	O	I-protein
,	NN	O	O
p50/RelA	NN	O	B-protein
,	NN	O	O
is	NN	O	O
induced	NN	O	O
in	NN	O	O
phagocytosing	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

Under	NN	O	O
these	NN	O	O
conditions	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
rapid	NN	O	O
and	NN	O	O
transient	NN	O	O
response	NN	O	O
,	NN	O	O
reaching	NN	O	O
a	NN	O	O
maximum	NN	O	O
by	NN	O	O
10-15	NN	O	O
min	NN	O	O
,	NN	O	O
and	NN	O	O
returning	NN	O	O
to	NN	O	O
near-basal	NN	O	O
levels	NN	O	O
by	NN	O	O
30	NN	O	O
min	NN	O	O
.	NN	O	O

In	NN	O	O
neutrophils	NN	O	O
undergoing	NN	O	O
the	NN	O	O
phagocytosis	NN	O	O
of	NN	O	O
opsonized	NN	O	O
yeasts	NN	O	O
,	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
paralleled	NN	O	O
by	NN	O	O
a	NN	O	O
decline	NN	O	O
in	NN	O	O
immunoreactive	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
protein	NN	O	O
levels	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
cellular	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
-alpha	NN	O	O
pool	NN	O	O
was	NN	O	O
replenished	NN	O	O
by	NN	O	O
30	NN	O	O
min	NN	O	O
,	NN	O	O
in	NN	O	O
agreement	NN	O	O
with	NN	O	O
our	NN	O	O
gel	NN	O	O
shift	NN	O	O
data	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
could	NN	O	O
constitute	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
whereby	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
kappa	NN	O	B-DNA
B-responsive	NN	O	I-DNA
genes	NN	O	I-DNA
is	NN	O	O
enhanced	NN	O	O
in	NN	O	O
phagocytosing	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
our	NN	O	O
knowledge	NN	O	O
,	NN	O	O
this	NN	O	O
represents	NN	O	O
the	NN	O	O
first	NN	O	O
demonstration	NN	O	O
that	NN	O	O
phagocytic	NN	O	O
stimuli	NN	O	O
can	NN	O	O
induce	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
Cryptococcus	NN	O	O
neoformans	NN	O	O
and	NN	O	O
Candida	NN	O	O
albicans	NN	O	O
.	NN	O	O

Because	NN	O	O
candidiasis	NN	O	O
and	NN	O	O
cryptococcosis	NN	O	O
are	NN	O	O
common	NN	O	O
in	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
-infected	NN	O	O
persons	NN	O	O
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
Cryptococcus	NN	O	O
neoformans	NN	O	O
and	NN	O	O
Candida	NN	O	O
albicans	NN	O	O
on	NN	O	O
HIV	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
examined	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
latently	NN	O	O
HIV-infected	NN	O	B-cell_line
myelomonocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
OM-10.1	NN	O	O
with	NN	O	O
C.	NN	O	O
neoformans	NN	O	O
and	NN	O	O
C.	NN	O	O
albicans	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
pooled	NN	O	O
human	NN	O	O
serum	NN	O	O
caused	NN	O	O
a	NN	O	O
ratio-dependent	NN	O	O
increase	NN	O	O
in	NN	O	O
HIV	NN	O	O
production	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
HIV	NN	O	O
by	NN	O	O
C.	NN	O	O
neoformans	NN	O	O
was	NN	O	O
enhanced	NN	O	O
by	NN	O	O
anti-capsular	NN	O	B-protein
antibody	NN	O	I-protein
,	NN	O	O
while	NN	O	O
induction	NN	O	O
by	NN	O	O
both	NN	O	O
organisms	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
anti-TNF-alpha	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

In	NN	O	O
THP-1	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
HIV	NN	O	B-DNA
plasmid	NN	O	I-DNA
constructs	NN	O	I-DNA
,	NN	O	O
both	NN	O	O
organisms	NN	O	O
induced	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
that	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
intact	NN	O	O
NF-kappaB	NN	O	B-DNA
binding	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
C.	NN	O	O
neoformans	NN	O	O
and	NN	O	O
C.	NN	O	O
albicans	NN	O	O
enhance	NN	O	O
HIV	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
a	NN	O	O
TNF-alpha-	NN	O	O
and	NN	O	O
NF-kappaB-dependent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

In	NN	O	O
HIV-infected	NN	O	O
patients	NN	O	O
,	NN	O	O
such	NN	O	O
enhancement	NN	O	O
may	NN	O	O
further	NN	O	O
impair	NN	O	O
host	NN	O	O
immunity	NN	O	O
and	NN	O	O
could	NN	O	O
accelerate	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
HIV	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
correlate	NN	O	O
with	NN	O	O
syncytium-forming	NN	O	O
capacity	NN	O	O
of	NN	O	O
8e51	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
expressing	NN	O	O
a	NN	O	O
defective	NN	O	O
HIV	NN	O	O
virus	NN	O	O
.	NN	O	O

The	NN	O	O
double	NN	O	B-DNA
NF-kappaB	NN	O	I-DNA
site	NN	O	I-DNA
identified	NN	O	O
in	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus-1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
be	NN	O	O
necessary	NN	O	O
for	NN	O	O
efficient	NN	O	O
viral	NN	O	O
transcription	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
present	NN	O	O
the	NN	O	O
characterisation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
subunits	NN	O	I-protein
engaged	NN	O	O
in	NN	O	O
complexes	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
NF-kappaB	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_line
8e51	NN	O	I-cell_line
T-cells	NN	O	I-cell_line
,	NN	O	O
that	NN	O	O
harbour	NN	O	O
a	NN	O	O
defective	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

At	NN	O	O
least	NN	O	O
four	NN	O	O
different	NN	O	O
specific	NN	O	O
NF-kappaB	NN	O	B-protein
complexes	NN	O	I-protein
are	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

With	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
specific	NN	O	B-protein
antibodies	NN	O	I-protein
we	NN	O	O
have	NN	O	O
determined	NN	O	O
the	NN	O	O
composition	NN	O	O
of	NN	O	O
each	NN	O	O
complex	NN	O	O
using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
show	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
several	NN	O	O
NF-kappaB	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
,	NN	O	I-protein
with	NN	O	O
the	NN	O	O
transactivating	NN	O	O
RelA	NN	O	B-protein
being	NN	O	O
engaged	NN	O	O
in	NN	O	O
multiple	NN	O	O
complexes	NN	O	O
.	NN	O	O

The	NN	O	O
importance	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
viral	NN	O	O
functions	NN	O	O
has	NN	O	O
been	NN	O	O
established	NN	O	O
comparing	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	O
DNA-binding	NN	O	O
complexes	NN	O	O
with	NN	O	O
syncytia-forming	NN	O	O
activity	NN	O	O
of	NN	O	O
8e51	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
8e51	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
had	NN	O	O
almost	NN	O	O
lost	NN	O	O
their	NN	O	O
syncytia-forming	NN	O	O
capacity	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
contain	NN	O	O
at	NN	O	O
least	NN	O	O
10	NN	O	O
times	NN	O	O
less	NN	O	O
active	NN	O	O
NF-kappaB	NN	O	B-protein
DNA-binding	NN	O	I-protein
complex	NN	O	I-protein
than	NN	O	O
the	NN	O	O
actively	NN	O	B-cell_type
fusing	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
correlation	NN	O	O
is	NN	O	O
specific	NN	O	O
as	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
did	NN	O	O
not	NN	O	O
change	NN	O	O
.	NN	O	O

-DOCSTART-	O

c-Myb	NN	O	B-DNA
and	NN	O	O
Ets	NN	O	B-protein
proteins	NN	O	I-protein
synergize	NN	O	O
to	NN	O	O
overcome	NN	O	O
transcriptional	NN	O	O
repression	NN	O	O
by	NN	O	O
ZEB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
Zfh	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
zinc	NN	O	B-protein
finger/homeodomain	NN	O	I-protein
proteins	NN	O	I-protein
was	NN	O	O
first	NN	O	O
identified	NN	O	O
in	NN	O	O
Drosophila	NN	O	O
where	NN	O	O
it	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
differentiation	NN	O	O
of	NN	O	O
tissues	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
and	NN	O	O
muscle	NN	O	O
.	NN	O	O

ZEB	NN	O	B-protein
,	NN	O	O
a	NN	O	O
vertebrate	NN	O	O
homolog	NN	O	O
of	NN	O	O
Zfh-1	NN	O	B-protein
,	NN	O	O
binds	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
E	NN	O	B-DNA
boxes	NN	O	I-DNA
and	NN	O	O
blocks	NN	O	O
myogenesis	NN	O	O
through	NN	O	O
transcriptional	NN	O	O
repression	NN	O	O
of	NN	O	O
muscle	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
present	NN	O	O
evidence	NN	O	O
here	NN	O	O
that	NN	O	O
ZEB	NN	O	B-protein
also	NN	O	O
has	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
controlling	NN	O	O
hematopoietic	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

Two	NN	O	O
families	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
normal	NN	O	O
hematopoiesis	NN	O	O
are	NN	O	O
c-Myb	NN	O	B-protein
and	NN	O	O
Ets	NN	O	B-protein
.	NN	O	O

These	NN	O	O
factors	NN	O	O
act	NN	O	O
synergistically	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
synergy	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
transcription	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
several	NN	O	O
important	NN	O	O
hematopoietic	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

ZEB	NN	O	B-protein
blocks	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
c-Myb	NN	O	B-protein
and	NN	O	O
Ets	NN	O	B-protein
individually	NN	O	O
,	NN	O	O
but	NN	O	O
together	NN	O	O
the	NN	O	O
factors	NN	O	O
synergize	NN	O	O
to	NN	O	O
resist	NN	O	O
this	NN	O	O
repression	NN	O	O
.	NN	O	O

Such	NN	O	O
repression	NN	O	O
imposes	NN	O	O
a	NN	O	O
requirement	NN	O	O
for	NN	O	O
both	NN	O	O
c-Myb	NN	O	B-protein
and	NN	O	O
Ets	NN	O	B-protein
for	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
,	NN	O	O
providing	NN	O	O
one	NN	O	O
explanation	NN	O	O
for	NN	O	O
why	NN	O	O
synergy	NN	O	O
between	NN	O	O
these	NN	O	O
factors	NN	O	O
is	NN	O	O
important	NN	O	O
.	NN	O	O

The	NN	O	O
balance	NN	O	O
between	NN	O	O
repression	NN	O	O
by	NN	O	O
ZEB	NN	O	B-protein
and	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
c-Myb	NN	O	B-protein
/Ets	NN	O	B-protein
provides	NN	O	O
a	NN	O	O
flexible	NN	O	O
regulatory	NN	O	O
mechanism	NN	O	O
for	NN	O	O
controlling	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
one	NN	O	O
target	NN	O	O
of	NN	O	O
this	NN	O	O
positive/negative	NN	O	O
regulation	NN	O	O
in	NN	O	O
vivo	NN	O	O
is	NN	O	O
the	NN	O	O
alpha4	NN	O	B-protein
integrin	NN	O	I-protein
,	NN	O	O
which	NN	O	O
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
normal	NN	O	O
hematopoiesis	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
mature	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Comparison	NN	O	O
of	NN	O	O
the	NN	O	O
transactivation	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
Stat5	NN	O	B-protein
and	NN	O	O
Stat6	NN	O	B-protein
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
mammary	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Stat	NN	O	B-protein
(	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
)	NN	O	O
and	NN	O	O
Jak	NN	O	B-protein
(	NN	O	O
Janus	NN	O	B-protein
kinases	NN	O	I-protein
)	NN	O	O
proteins	NN	O	O
are	NN	O	O
central	NN	O	O
components	NN	O	O
in	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
and	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

They	NN	O	O
are	NN	O	O
rapidly	NN	O	O
activated	NN	O	O
by	NN	O	O
various	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
hormones	NN	O	O
,	NN	O	O
and	NN	O	O
growth	NN	O	O
factors	NN	O	O
.	NN	O	O

Upon	NN	O	O
ligand	NN	O	O
binding	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
dimerization	NN	O	O
,	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
residues	NN	O	O
by	NN	O	O
Jak	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

Activated	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
form	NN	O	O
homo-	NN	O	O
or	NN	O	O
heterodimers	NN	O	O
,	NN	O	O
translocate	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
and	NN	O	O
induce	NN	O	O
transcription	NN	O	O
from	NN	O	O
responsive	NN	O	O
genes	NN	O	O
.	NN	O	O

Stat5	NN	O	B-protein
and	NN	O	O
Stat6	NN	O	B-protein
are	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
active	NN	O	O
in	NN	O	O
mammary	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
immune	NN	O	O
cells	NN	O	O
.	NN	O	O

Prolactin	NN	O	O
activates	NN	O	O
Stat5	NN	O	B-protein
,	NN	O	O
and	NN	O	O
interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
activates	NN	O	O
Stat6	NN	O	B-protein
.	NN	O	O

Both	NN	O	O
cytokines	NN	O	B-protein
are	NN	O	O
able	NN	O	O
to	NN	O	O
stimulate	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
survival	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
transactivation	NN	O	O
potential	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
and	NN	O	O
found	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
restricted	NN	O	O
to	NN	O	O
lymphocytes	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
was	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
HC11	NN	O	B-cell_line
mammary	NN	O	I-cell_line
epithelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
Stat6	NN	O	B-protein
activation	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
beta-casein	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
this	NN	O	O
promoter	NN	O	O
was	NN	O	O
confirmed	NN	O	O
in	NN	O	O
COS7	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
further	NN	O	O
enhance	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
gene	NN	O	O
transcription	NN	O	O
through	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
.	NN	O	O

Deletion	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
carboxyl-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
Stat6	NN	O	B-protein
and	NN	O	O
recombination	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
with	NN	O	O
a	NN	O	O
heterologous	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
allowed	NN	O	O
the	NN	O	O
delimitation	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
transactivation	NN	O	O
domain	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
.	NN	O	O

The	NN	O	O
potencies	NN	O	O
of	NN	O	O
the	NN	O	O
transactivation	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
Stat5	NN	O	B-protein
,	NN	O	O
Stat6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
viral	NN	O	B-protein
protein	NN	O	I-protein
VP16	NN	O	I-protein
were	NN	O	O
compared	NN	O	O
.	NN	O	O

Stat6	NN	O	B-protein
had	NN	O	O
a	NN	O	O
transactivation	NN	O	O
domain	NN	O	O
which	NN	O	O
was	NN	O	O
about	NN	O	O
10-fold	NN	O	O
stronger	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
Stat5	NN	O	B-protein
.	NN	O	O

In	NN	O	O
pre-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
Ba/F3	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
transactivation	NN	O	O
domain	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
was	NN	O	O
IL-4	NN	O	B-protein
regulated	NN	O	O
,	NN	O	O
independently	NN	O	O
from	NN	O	O
its	NN	O	O
DNA	NN	O	O
binding	NN	O	O
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

ETS1	NN	O	B-protein
,	NN	O	O
NFkappaB	NN	O	B-protein
and	NN	O	O
AP1	NN	O	B-protein
synergistically	NN	O	O
transactivate	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
helper	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
results	NN	O	O
in	NN	O	O
coordinate	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
involved	NN	O	O
in	NN	O	O
differentiation	NN	O	O
,	NN	O	O
proliferation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
haematopoietic	NN	O	O
system	NN	O	O
.	NN	O	O

Granulocyte-macrophage	NN	O	B-protein
colony	NN	O	I-protein
stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
is	NN	O	O
one	NN	O	O
such	NN	O	O
cytokine	NN	O	B-protein
,	NN	O	O
whose	NN	O	O
increased	NN	O	O
expression	NN	O	O
results	NN	O	O
mostly	NN	O	O
from	NN	O	O
increases	NN	O	O
in	NN	O	O
transcription	NN	O	O
.	NN	O	O

Cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
with	NN	O	O
NFkappaB	NN	O	B-DNA
,	NN	O	I-DNA
AP1	NN	O	I-DNA
and	NN	O	I-DNA
ETS-like	NN	O	I-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
are	NN	O	O
important	NN	O	O
or	NN	O	O
essential	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
following	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

ETS1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
ETS	NN	O	B-protein
family	NN	O	I-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
ETS1	NN	O	B-protein
can	NN	O	O
transactivate	NN	O	O
GM-CSF	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
only	NN	O	O
after	NN	O	O
the	NN	O	O
cells	NN	O	O
have	NN	O	O
been	NN	O	O
stimulated	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
PMA	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
,	NN	O	O
agents	NN	O	O
that	NN	O	O
mimic	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Thus	NN	O	O
we	NN	O	O
proposed	NN	O	O
that	NN	O	O
ETS1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
expressed	NN	O	O
constitutively	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
may	NN	O	O
act	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
PMA/ionomycin	NN	O	B-protein
inducible	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
ETS1	NN	O	B-protein
can	NN	O	O
transactivate	NN	O	O
a	NN	O	O
GM-CSF	NN	O	B-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
in	NN	O	O
unstimulated	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
providing	NN	O	O
that	NN	O	O
either	NN	O	O
NFkappaB	NN	O	B-protein
or	NN	O	O
AP1	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
supplied	NN	O	O
by	NN	O	O
co-transfection	NN	O	O
.	NN	O	O

We	NN	O	O
confirm	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
endogenous	NN	O	O
NFkappaB	NN	O	B-protein
and	NN	O	O
AP1	NN	O	B-protein
is	NN	O	O
induced	NN	O	O
following	NN	O	O
PMA/ionomycin	NN	O	O
treatment	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Transactivation	NN	O	O
by	NN	O	O
ETS1	NN	O	B-protein
,	NN	O	O
NFkappaB	NN	O	B-protein
and	NN	O	O
AP1	NN	O	B-protein
is	NN	O	O
synergistic	NN	O	O
,	NN	O	O
and	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
individual	NN	O	O
binding	NN	O	O
sites	NN	O	O
reveals	NN	O	O
that	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
are	NN	O	O
interdependent	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
constitutive	NN	O	O
ETS1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
inducible	NN	O	O
NFkappaB	NN	O	B-protein
and	NN	O	O
AP1	NN	O	B-protein
,	NN	O	O
cooperate	NN	O	O
as	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
higher	NN	O	O
order	NN	O	O
transcriptional	NN	O	B-protein
complex	NN	O	I-protein
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Abnormal	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
development	NN	O	O
induced	NN	O	O
by	NN	O	O
targeted	NN	O	O
overexpression	NN	O	O
of	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

A	NN	O	O
role	NN	O	O
in	NN	O	O
thymic	NN	O	O
maturation	NN	O	O
for	NN	O	O
factors	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
family	NN	O	I-protein
has	NN	O	O
long	NN	O	O
been	NN	O	O
suspected	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
yet	NN	O	O
proven	NN	O	O
.	NN	O	O

Transgenic	NN	O	O
mice	NN	O	O
with	NN	O	O
a	NN	O	O
lymphocyte-specific	NN	O	O
defect	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
were	NN	O	O
produced	NN	O	O
by	NN	O	O
targeted	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
thymic	NN	O	O
cellularity	NN	O	O
of	NN	O	O
these	NN	O	O
mice	NN	O	O
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
.	NN	O	O

The	NN	O	O
proportion	NN	O	O
of	NN	O	O
mature	NN	O	O
,	NN	O	O
TCRhigh	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
alphabeta	NN	O	B-cell_type
lineage	NN	O	I-cell_type
was	NN	O	O
reduced	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
remaining	NN	O	O
TCRhigh	NN	O	B-cell_type
population	NN	O	I-cell_type
contained	NN	O	O
an	NN	O	O
unusually	NN	O	O
high	NN	O	O
proportion	NN	O	O
of	NN	O	O
double-positive	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
defect	NN	O	O
in	NN	O	O
maturation	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
transgene	NN	O	O
dose-dependent	NN	O	O
reduction	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
with	NN	O	O
the	NN	O	O
CD8	NN	O	B-cell_type
lineage	NN	O	I-cell_type
being	NN	O	O
more	NN	O	O
severely	NN	O	O
affected	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
direct	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
late	NN	O	O
stages	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
development	NN	O	O
,	NN	O	O
coincident	NN	O	O
with	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	O
selection	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD40	NN	O	B-protein
is	NN	O	O
a	NN	O	O
functional	NN	O	O
activation	NN	O	B-protein
antigen	NN	O	I-protein
and	NN	O	O
B7-independent	NN	O	B-protein
T	NN	O	I-protein
cell	NN	O	I-protein
costimulatory	NN	O	I-protein
molecule	NN	O	I-protein
on	NN	O	O
normal	NN	O	O
human	NN	O	B-cell_type
lung	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

CD40	NN	O	B-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
signaling	NN	O	O
and	NN	O	O
activation	NN	O	O
Ag	NN	O	O
found	NN	O	O
on	NN	O	O
certain	NN	O	O
bone	NN	O	B-cell_type
marrow-derived	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
CD40	NN	O	B-protein
also	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
by	NN	O	O
mesenchymal	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
including	NN	O	O
human	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

Little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
in	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
current	NN	O	O
study	NN	O	O
investigates	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
CD40	NN	O	B-protein
expressed	NN	O	O
on	NN	O	O
lung	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
is	NN	O	O
an	NN	O	O
activation	NN	O	O
structure	NN	O	O
and	NN	O	O
mechanism	NN	O	O
for	NN	O	O
interaction	NN	O	O
with	NN	O	O
hemopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Communication	NN	O	O
between	NN	O	O
resident	NN	O	O
tissue	NN	O	O
fibroblasts	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
normal	NN	O	O
wound	NN	O	O
healing	NN	O	O
,	NN	O	O
and	NN	O	O
can	NN	O	O
be	NN	O	O
pathologic	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
tissue	NN	O	O
fibrosis	NN	O	O
.	NN	O	O

Signaling	NN	O	O
through	NN	O	O
CD40	NN	O	B-protein
with	NN	O	O
soluble	NN	O	O
CD40	NN	O	B-protein
ligand	NN	O	O
stimulated	NN	O	O
fibroblast	NN	O	O
activation	NN	O	O
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
mobilization	NN	O	O
of	NN	O	O
nuclear	NN	O	O
factor-kappaB	NN	O	O
and	NN	O	O
by	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
proinflammatory	NN	O	O
and	NN	O	O
chemoattractant	NN	O	O
cytokines	NN	O	B-protein
IL-6	NN	O	B-protein
and	NN	O	O
IL-8	NN	O	B-protein
.	NN	O	O

IFN-gamma-primed	NN	O	B-cell_line
lung	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
costimulate	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
utilizing	NN	O	O
CD40	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
well-studied	NN	O	O
costimulatory	NN	O	B-protein
molecules	NN	O	I-protein
B7-1	NN	O	B-protein
and	NN	O	O
B7-2	NN	O	B-protein
.	NN	O	O

Data	NN	O	O
reported	NN	O	O
herein	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
cognate	NN	O	O
interactions	NN	O	O
between	NN	O	O
tissue	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
and	NN	O	O
infiltrating	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
,	NN	O	O
via	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
/CD40L	NN	O	B-protein
pathway	NN	O	O
,	NN	O	O
augment	NN	O	O
inflammation	NN	O	O
and	NN	O	O
may	NN	O	O
promote	NN	O	O
fibrogenesis	NN	O	O
by	NN	O	O
activating	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

-DOCSTART-	O

Biphasic	NN	O	O
control	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
triggering	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
antigens	NN	O	I-protein
expressed	NN	O	O
on	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
following	NN	O	O
the	NN	O	O
triggering	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
antigens	NN	O	I-protein
by	NN	O	O
mAb	NN	O	B-protein
L243	NN	O	I-protein
has	NN	O	O
been	NN	O	O
studied	NN	O	O
at	NN	O	O
various	NN	O	O
times	NN	O	O
in	NN	O	O
Raji	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
strong	NN	O	O
increase	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
after	NN	O	O
triggering	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
antigens	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
TNF-alpha-activity	NN	O	B-protein
neutralizing	NN	O	I-protein
antibodies	NN	O	I-protein
,	NN	O	O
the	NN	O	O
authors	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
depend	NN	O	O
,	NN	O	O
at	NN	O	O
later	NN	O	O
time	NN	O	O
point	NN	O	O
,	NN	O	O
on	NN	O	O
an	NN	O	O
autocrine	NN	O	O
effect	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
secreted	NN	O	O
following	NN	O	O
triggering	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
antigens	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
TNF-alpha	NN	O	B-protein
independent	NN	O	O
in	NN	O	O
the	NN	O	O
early	NN	O	O
time	NN	O	O
point	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
upregulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
triggering	NN	O	O
of	NN	O	O
selected	NN	O	O
epitopes	NN	O	B-protein
of	NN	O	O
HLA-DR	NN	O	B-protein
antigens	NN	O	I-protein
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
mAb	NN	O	B-protein
L243	NN	O	I-protein
but	NN	O	O
not	NN	O	O
the	NN	O	O
staphylococcal	NN	O	B-protein
superantigens	NN	O	I-protein
,	NN	O	O
staphylococcal	NN	O	B-protein
exotoxin	NN	O	I-protein
toxic	NN	O	I-protein
shock	NN	O	I-protein
syndrome	NN	O	I-protein
toxin-I	NN	O	I-protein
or	NN	O	O
staphylococcal	NN	O	B-protein
enterotoxin	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
regulate	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
inosine-5'-monophosphate	NN	O	B-protein
dehydrogenase	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Role	NN	O	O
for	NN	O	O
a	NN	O	O
novel	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
palindromic	NN	O	I-DNA
octamer	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
human	NN	O	B-protein
inosine-	NN	O	I-protein
5'-monophosphate	NN	O	I-protein
dehydrogenase	NN	O	I-protein
(	NN	O	I-protein
IMPDH	NN	O	I-protein
)	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
,	NN	O	O
an	NN	O	O
enzyme	NN	O	B-protein
catalyzing	NN	O	O
the	NN	O	O
rate-limiting	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
guanine	NN	O	O
nucleotides	NN	O	O
,	NN	O	O
is	NN	O	O
increased	NN	O	O
more	NN	O	O
than	NN	O	O
10-fold	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
5'-regulatory	NN	O	B-DNA
sequences	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

DNase	NN	O	B-protein
I	NN	O	I-protein
mapping	NN	O	O
of	NN	O	O
genomic	NN	O	B-DNA
DNA	NN	O	I-DNA
identified	NN	O	O
a	NN	O	O
hypersensitive	NN	O	B-DNA
element	NN	O	I-DNA
near	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Fine	NN	O	O
mapping	NN	O	O
by	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprinting	NN	O	O
demonstrated	NN	O	O
five	NN	O	O
transcription	NN	O	O
factor	NN	O	O
binding	NN	O	O
sites	NN	O	O
that	NN	O	O
are	NN	O	O
occupied	NN	O	O
in	NN	O	O
both	NN	O	O
resting	NN	O	O
and	NN	O	O
activated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
;	NN	O	O
these	NN	O	O
are	NN	O	O
tandem	NN	O	B-DNA
CRE	NN	O	I-DNA
motifs	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
an	NN	O	O
overlapping	NN	O	O
Egr-1/Sp1	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
novel	NN	O	O
palindromic	NN	O	B-DNA
octamer	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
POS	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

The	NN	O	O
tandem	NN	O	O
CRE	NN	O	B-DNA
and	NN	O	I-DNA
POS	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
of	NN	O	O
major	NN	O	O
functional	NN	O	O
importance	NN	O	O
as	NN	O	O
judged	NN	O	O
by	NN	O	O
mutational	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analyses	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IMPDH	NN	O	I-DNA
type	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
predominantly	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
ATF-2	NN	O	B-protein
and	NN	O	O
an	NN	O	O
as	NN	O	O
yet	NN	O	O
unidentified	NN	O	O
POS-binding	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Additional	NN	O	O
major	NN	O	O
protein-DNA	NN	O	O
interactions	NN	O	O
do	NN	O	O
not	NN	O	O
occur	NN	O	O
within	NN	O	O
the	NN	O	O
promoter	NN	O	O
region	NN	O	O
after	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
requirement	NN	O	O
for	NN	O	O
additional	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
and/or	NN	O	O
post-translational	NN	O	O
modifications	NN	O	O
of	NN	O	O
pre-bound	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
to	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
observed	NN	O	O
increase	NN	O	O
in	NN	O	O
IMPDH	NN	O	O
type	NN	O	O
II	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Distinct	NN	O	O
mechanisms	NN	O	O
for	NN	O	O
N-acetylcysteine	NN	O	O
inhibition	NN	O	O
of	NN	O	O
cytokine-induced	NN	O	B-protein
E-selectin	NN	O	I-protein
and	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
N-acetyl-L-cysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
well-characterized	NN	O	O
,	NN	O	O
thiol-containing	NN	O	O
antioxidant	NN	O	O
,	NN	O	O
on	NN	O	O
agonist-induced	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
EC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

NAC	NN	O	O
inhibited	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
)	NN	O	O
-induced	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
basal	NN	O	O
,	NN	O	O
adhesion	NN	O	O
with	NN	O	O
50	NN	O	O
%	NN	O	O
inhibition	NN	O	O
at	NN	O	O
approximately	NN	O	O
20	NN	O	O
mM	NN	O	O
.	NN	O	O

Monocytic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
to	NN	O	O
EC	NN	O	B-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
alpha-thrombin	NN	O	B-protein
,	NN	O	O
or	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
was	NN	O	O
similarly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
NAC	NN	O	O
.	NN	O	O

Unlike	NN	O	O
published	NN	O	O
studies	NN	O	O
with	NN	O	O
pyrrolidinedithiocarbamate	NN	O	O
,	NN	O	O
which	NN	O	O
specifically	NN	O	O
inhibited	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
NAC	NN	O	O
inhibited	NN	O	O
IL-1	NN	O	B-RNA
beta-induced	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
E-selectin	NN	O	B-protein
and	NN	O	O
VCAM-1	NN	O	B-protein
.	NN	O	O

NAC	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
half-life	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-RNA
or	NN	O	I-RNA
VCAM-1	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Although	NN	O	O
NAC	NN	O	O
reduced	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
activation	NN	O	O
in	NN	O	O
EC	NN	O	B-cell_type
as	NN	O	O
measured	NN	O	O
by	NN	O	O
gel-shift	NN	O	O
assays	NN	O	O
using	NN	O	O
an	NN	O	O
oligonucleotide	NN	O	B-DNA
probe	NN	O	I-DNA
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
VCAM-1	NN	O	B-DNA
gene	NN	O	I-DNA
(	NN	O	O
VCAM-NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
had	NN	O	O
no	NN	O	O
appreciable	NN	O	O
effect	NN	O	O
when	NN	O	O
an	NN	O	O
oligomer	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
E-selectin	NN	O	B-DNA
gene	NN	O	I-DNA
(	NN	O	O
E-selectin	NN	O	B-protein
-NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
was	NN	O	O
used	NN	O	O
.	NN	O	O

Because	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
redox	NN	O	O
sensitive	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
NAC	NN	O	O
on	NN	O	O
the	NN	O	O
EC	NN	O	B-cell_type
redox	NN	O	O
environment	NN	O	O
.	NN	O	O

NAC	NN	O	O
caused	NN	O	O
an	NN	O	O
expected	NN	O	O
dramatic	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
reduced	NN	O	O
glutathione	NN	O	O
(	NN	O	O
GSH	NN	O	O
)	NN	O	O
levels	NN	O	O
in	NN	O	O
EC	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
whereas	NN	O	O
the	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
to	NN	O	O
the	NN	O	O
VCAM-NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
oligomer	NN	O	I-DNA
peaked	NN	O	O
at	NN	O	O
a	NN	O	O
GSH-to-oxidized	NN	O	O
glutathione	NN	O	O
(	NN	O	O
GSSG	NN	O	O
)	NN	O	O
ratio	NN	O	O
of	NN	O	O
approximately	NN	O	O
200	NN	O	O
and	NN	O	O
decreased	NN	O	O
at	NN	O	O
higher	NN	O	O
ratios	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
E-selectin-NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
oligomer	NN	O	I-DNA
appeared	NN	O	O
relatively	NN	O	O
unaffected	NN	O	O
even	NN	O	O
at	NN	O	O
ratios	NN	O	O
>	NN	O	O
400	NN	O	O
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
those	NN	O	O
achieved	NN	O	O
in	NN	O	O
EC	NN	O	B-cell_type
treated	NN	O	O
with	NN	O	O
40	NN	O	O
mM	NN	O	O
NAC	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
its	NN	O	O
consensus	NN	O	O
sequences	NN	O	O
in	NN	O	O
the	NN	O	O
VCAM-1	NN	O	B-protein
and	NN	O	O
E-selectin	NN	O	B-DNA
gene	NN	O	I-DNA
exhibits	NN	O	O
marked	NN	O	O
differences	NN	O	O
in	NN	O	O
redox	NN	O	O
sensitivity	NN	O	O
,	NN	O	O
allowing	NN	O	O
for	NN	O	O
differential	NN	O	O
gene	NN	O	O
expression	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
same	NN	O	O
transcription	NN	O	O
factor	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
NAC	NN	O	O
increases	NN	O	O
the	NN	O	O
GSH-to-GSSG	NN	O	O
ratio	NN	O	O
within	NN	O	O
the	NN	O	O
EC	NN	O	B-cell_type
suggesting	NN	O	O
one	NN	O	O
possible	NN	O	O
mechanism	NN	O	O
through	NN	O	O
which	NN	O	O
this	NN	O	O
antioxidant	NN	O	O
inhibits	NN	O	O
agonist-induced	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
EC	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Opposite	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
acute	NN	O	B-protein
promyelocytic	NN	O	I-protein
leukemia	NN	O	I-protein
PML-retinoic	NN	O	I-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
and	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
fusion	NN	O	I-protein
proteins	NN	O	I-protein
on	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
signalling	NN	O	O
.	NN	O	O

Fusion	NN	O	O
proteins	NN	O	O
involving	NN	O	O
the	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
receptor	NN	O	B-protein
alpha	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
PML	NN	O	B-protein
or	NN	O	O
PLZF	NN	O	B-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
are	NN	O	O
the	NN	O	O
genetic	NN	O	O
markers	NN	O	O
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemias	NN	O	O
(	NN	O	O
APLs	NN	O	O
)	NN	O	O
.	NN	O	O

APLs	NN	O	O
with	NN	O	O
the	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
or	NN	O	O
the	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
are	NN	O	O
phenotypically	NN	O	O
indistinguishable	NN	O	O
except	NN	O	O
that	NN	O	O
they	NN	O	O
differ	NN	O	O
in	NN	O	O
their	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
-induced	NN	O	O
differentiation	NN	O	O
:	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
blasts	NN	O	O
are	NN	O	O
sensitive	NN	O	O
to	NN	O	O
RA	NN	O	O
and	NN	O	O
patients	NN	O	O
enter	NN	O	O
disease	NN	O	O
remission	NN	O	O
after	NN	O	O
RA	NN	O	O
treatment	NN	O	O
,	NN	O	O
while	NN	O	O
patients	NN	O	O
with	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
do	NN	O	O
not	NN	O	O
.	NN	O	O

We	NN	O	O
here	NN	O	O
report	NN	O	O
that	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
like	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
blocks	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
of	NN	O	O
hematopoietic	NN	O	O
precursor	NN	O	O
cell	NN	O	O
lines	NN	O	O
(	NN	O	O
U937	NN	O	B-cell_line
and	NN	O	O
HL-60	NN	O	B-cell_line
)	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
different	NN	O	O
stimuli	NN	O	O
(	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
,	NN	O	O
transforming	NN	O	O
growth	NN	O	O
factor	NN	O	O
beta1	NN	O	O
,	NN	O	O
and	NN	O	O
dimethyl	NN	O	O
sulfoxide	NN	O	O
)	NN	O	O
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
increases	NN	O	O
RA	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
restores	NN	O	O
RA	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
RA-resistant	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
have	NN	O	O
similar	NN	O	O
RA	NN	O	O
binding	NN	O	O
affinities	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
iv	NN	O	O
)	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
enhances	NN	O	O
the	NN	O	O
RA	NN	O	O
response	NN	O	O
of	NN	O	O
RA	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
(	NN	O	O
those	NN	O	O
for	NN	O	O
RAR	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
RAR	NN	O	B-protein
gamma	NN	O	I-protein
,	NN	O	O
and	NN	O	O
transglutaminase	NN	O	B-protein
type	NN	O	I-protein
II	NN	O	I-protein
[	NN	O	O
TGase	NN	O	B-protein
]	NN	O	O
)	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
while	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
has	NN	O	O
either	NN	O	O
no	NN	O	O
effect	NN	O	O
(	NN	O	O
RAR	NN	O	B-protein
beta	NN	O	I-protein
)	NN	O	O
or	NN	O	O
an	NN	O	O
inhibitory	NN	O	O
activity	NN	O	O
(	NN	O	O
RAR	NN	O	B-protein
gamma	NN	O	I-protein
and	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
TGase	NN	O	I-protein
)	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
have	NN	O	O
similar	NN	O	O
(	NN	O	O
inhibitory	NN	O	O
)	NN	O	O
effects	NN	O	O
on	NN	O	O
RA-independent	NN	O	O
differentiation	NN	O	O
and	NN	O	O
opposite	NN	O	O
(	NN	O	O
stimulatory	NN	O	O
or	NN	O	O
inhibitory	NN	O	O
)	NN	O	O
effects	NN	O	O
on	NN	O	O
RA-dependent	NN	O	O
differentiation	NN	O	O
and	NN	O	O
that	NN	O	O
they	NN	O	O
behave	NN	O	O
in	NN	O	O
vivo	NN	O	O
as	NN	O	O
RA-dependent	NN	O	B-DNA
enhancers	NN	O	I-DNA
or	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
RA-responsive	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Their	NN	O	O
different	NN	O	O
activities	NN	O	O
on	NN	O	O
the	NN	O	O
RA	NN	O	O
signalling	NN	O	O
pathway	NN	O	O
might	NN	O	O
underlie	NN	O	O
the	NN	O	O
different	NN	O	O
responses	NN	O	O
of	NN	O	O
PML-RAR	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
APLs	NN	O	O
to	NN	O	O
RA	NN	O	O
treatment	NN	O	O
.	NN	O	O

The	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
contains	NN	O	O
an	NN	O	O
approximately	NN	O	O
120-amino-acid	NN	O	B-protein
N-terminal	NN	O	I-protein
motif	NN	O	I-protein
(	NN	O	O
called	NN	O	O
the	NN	O	O
POZ	NN	O	B-protein
domain	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
zinc	NN	O	O
finger	NN	O	O
proteins	NN	O	O
and	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
poxvirus	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
which	NN	O	O
mediates	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
the	NN	O	O
PLZF	NN	O	B-protein
POZ	NN	O	I-protein
domain	NN	O	I-protein
partially	NN	O	O
abrogated	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
on	NN	O	O
RA-induced	NN	O	O
differentiation	NN	O	O
and	NN	O	O
on	NN	O	O
RA-mediated	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
TGase	NN	O	I-protein
up-regulation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
POZ-mediated	NN	O	O
protein	NN	O	O
interactions	NN	O	O
might	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
of	NN	O	O
PLZF-RAR	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
human	NN	O	O
homologue	NN	O	O
of	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-protein
Toll	NN	O	I-protein
protein	NN	O	I-protein
signals	NN	O	O
activation	NN	O	O
of	NN	O	O
adaptive	NN	O	O
immunity	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Induction	NN	O	O
of	NN	O	O
the	NN	O	O
adaptive	NN	O	O
immune	NN	O	O
response	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
co-stimulatory	NN	O	B-protein
molecules	NN	O	I-protein
and	NN	O	O
cytokines	NN	O	B-protein
by	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
the	NN	O	O
initial	NN	O	O
induction	NN	O	O
of	NN	O	O
these	NN	O	O
signals	NN	O	O
upon	NN	O	O
infection	NN	O	O
are	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
proposed	NN	O	O
that	NN	O	O
their	NN	O	O
expression	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
the	NN	O	O
non-clonal	NN	O	O
,	NN	O	O
or	NN	O	O
innate	NN	O	O
,	NN	O	O
component	NN	O	O
of	NN	O	O
immunity	NN	O	O
that	NN	O	O
preceded	NN	O	O
in	NN	O	O
evolution	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
an	NN	O	O
adaptive	NN	O	O
immune	NN	O	O
system	NN	O	O
in	NN	O	O
vertebrates	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
the	NN	O	O
cloning	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-protein
homologue	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-protein
toll	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
Toll	NN	O	B-protein
)	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
innate	NN	O	O
immune	NN	O	O
response	NN	O	O
in	NN	O	O
adult	NN	O	O
Drosophila	NN	O	O
.	NN	O	O

Like	NN	O	O
Drosophila	NN	O	O
Toll	NN	O	B-protein
,	NN	O	O
human	NN	O	B-protein
Toll	NN	O	I-protein
is	NN	O	O
a	NN	O	O
type	NN	O	O
I	NN	O	O
transmembrane	NN	O	O
protein	NN	O	O
with	NN	O	O
an	NN	O	O
extracellular	NN	O	O
domain	NN	O	O
consisting	NN	O	O
of	NN	O	O
a	NN	O	O
leucine-rich	NN	O	B-protein
repeat	NN	O	I-protein
(	NN	O	I-protein
LRR	NN	O	I-protein
)	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	O
domain	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
interleukin	NN	O	I-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Both	NN	O	O
Drosophila	NN	O	O
Toll	NN	O	B-protein
and	NN	O	O
the	NN	O	O
IL-1	NN	O	O
receptor	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
signal	NN	O	O
through	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
constitutively	NN	O	O
active	NN	O	O
mutant	NN	O	O
of	NN	O	O
human	NN	O	B-protein
Toll	NN	O	I-protein
transfected	NN	O	O
into	NN	O	O
human	NN	O	O
cell	NN	O	O
lines	NN	O	O
can	NN	O	O
induce	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappaB-controlled	NN	O	B-DNA
genes	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
IL-1	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
co-stimulatory	NN	O	B-protein
molecule	NN	O	I-protein
B7.1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Role	NN	O	O
of	NN	O	O
ascorbate	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
by	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
in	NN	O	O
T-cells	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
first	NN	O	O
product	NN	O	O
of	NN	O	O
ascorbate	NN	O	O
oxidation	NN	O	O
,	NN	O	O
the	NN	O	O
ascorbate	NN	O	O
free	NN	O	O
radical	NN	O	O
(	NN	O	O
AFR	NN	O	O
)	NN	O	O
,	NN	O	O
acts	NN	O	O
in	NN	O	O
biological	NN	O	O
systems	NN	O	O
mainly	NN	O	O
as	NN	O	O
an	NN	O	O
oxidant	NN	O	O
,	NN	O	O
and	NN	O	O
through	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
redox	NN	O	O
system	NN	O	O
exerts	NN	O	O
different	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
ascorbate	NN	O	O
,	NN	O	O
AFR	NN	O	O
and	NN	O	O
dehydroascorbate	NN	O	O
(	NN	O	O
DHA	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
stimulated	NN	O	O
by	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
,	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
that	NN	O	O
ascorbate	NN	O	O
increases	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
to	NN	O	O
DNA	NN	O	O
in	NN	O	O
TNF-alpha-stimulated	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
ascorbate	NN	O	O
to	NN	O	O
enhance	NN	O	O
cytoplasmic	NN	O	O
inhibitory	NN	O	O
IkBalpha	NN	O	B-protein
protein	NN	O	I-protein
degradation	NN	O	O
correlates	NN	O	O
completely	NN	O	O
with	NN	O	O
its	NN	O	O
capacity	NN	O	O
to	NN	O	O
induce	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
and	NN	O	O
to	NN	O	O
potentiate	NN	O	O
NF-kappaB	NN	O	B-protein
-mediated	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
TNF-alpha-stimulated	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
cells	NN	O	O
stimulated	NN	O	O
with	NN	O	O
PMA	NN	O	O
plus	NN	O	O
ionomycin	NN	O	O
.	NN	O	O

AFR	NN	O	O
behaves	NN	O	O
like	NN	O	O
ascorbate	NN	O	O
,	NN	O	O
while	NN	O	O
DHA	NN	O	O
and	NN	O	O
ascorbate	NN	O	O
phosphate	NN	O	O
do	NN	O	O
not	NN	O	O
affect	NN	O	O
TNF-alpha	NN	O	B-protein
-mediated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
new	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
possible	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
electron-transport	NN	O	O
system	NN	O	O
at	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
by	NN	O	O
ascorbate	NN	O	O
or	NN	O	O
its	NN	O	O
free	NN	O	O
radical	NN	O	O
and	NN	O	O
redox-dependent	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
T-cells	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
cytomegalovirus	NN	O	O
induces	NN	O	O
interleukin-8	NN	O	B-protein
production	NN	O	O
by	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
,	NN	O	O
through	NN	O	O
acting	NN	O	O
concurrently	NN	O	O
on	NN	O	O
AP-1-	NN	O	B-DNA
and	NN	O	I-DNA
NF-kappaB-binding	NN	O	I-DNA
sites	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
interleukin-8	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Cytomegalovirus	NN	O	O
(	NN	O	O
CMV	NN	O	O
)	NN	O	O
infection	NN	O	O
induced	NN	O	O
interleukin-8	NN	O	B-protein
(	NN	O	O
IL-8	NN	O	B-protein
)	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
leading	NN	O	O
to	NN	O	O
IL-8	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

The	NN	O	O
functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
IL-8	NN	O	B-DNA
gene	NN	O	I-DNA
revealed	NN	O	O
that	NN	O	O
both	NN	O	O
AP-1-	NN	O	B-DNA
and	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
factor-binding	NN	O	I-DNA
elements	NN	O	I-DNA
were	NN	O	O
involved	NN	O	O
in	NN	O	O
conferring	NN	O	O
the	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
CMV	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
CMV	NN	O	O
induced	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
CMV	NN	O	O
activates	NN	O	O
these	NN	O	O
transcriptional	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
IL-8	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Construction	NN	O	O
and	NN	O	O
biological	NN	O	O
characterization	NN	O	O
of	NN	O	O
an	NN	O	O
interleukin-12	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
Flexi-12	NN	O	B-protein
)	NN	O	O
:	NN	O	O
delivery	NN	O	O
to	NN	O	O
acute	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
leukemic	NN	O	I-cell_type
blasts	NN	O	I-cell_type
using	NN	O	O
adeno-associated	NN	O	O
virus	NN	O	O
.	NN	O	O

Interleukin-12	NN	O	B-protein
(	NN	O	O
IL-12	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
that	NN	O	O
exhibits	NN	O	O
pleiotropic	NN	O	O
effects	NN	O	O
on	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
have	NN	O	O
promise	NN	O	O
for	NN	O	O
the	NN	O	O
immunotherapy	NN	O	O
of	NN	O	O
cancer	NN	O	O
.	NN	O	O

The	NN	O	O
combination	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	B-protein
costimulatory	NN	O	I-protein
molecule	NN	O	I-protein
B7.1	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
synergistic	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

By	NN	O	O
transfecting	NN	O	O
tumor	NN	O	O
cells	NN	O	O
with	NN	O	O
both	NN	O	O
IL-12	NN	O	B-DNA
and	NN	O	I-DNA
B7.1	NN	O	I-DNA
cDNAs	NN	O	I-DNA
,	NN	O	O
it	NN	O	O
may	NN	O	O
be	NN	O	O
possible	NN	O	O
to	NN	O	O
use	NN	O	O
these	NN	O	O
modified	NN	O	O
targets	NN	O	O
as	NN	O	O
vaccines	NN	O	O
.	NN	O	O

A	NN	O	O
major	NN	O	O
obstacle	NN	O	O
in	NN	O	O
designing	NN	O	O
a	NN	O	O
vector	NN	O	O
to	NN	O	O
deliver	NN	O	O
these	NN	O	O
genes	NN	O	O
results	NN	O	O
from	NN	O	O
the	NN	O	O
structure	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
.	NN	O	O

Functional	NN	O	O
IL-12	NN	O	B-protein
is	NN	O	O
a	NN	O	O
heterodimer	NN	O	B-protein
composed	NN	O	O
of	NN	O	O
two	NN	O	O
distinct	NN	O	O
subunits	NN	O	O
that	NN	O	O
are	NN	O	O
encoded	NN	O	O
by	NN	O	O
separate	NN	O	O
genes	NN	O	O
on	NN	O	O
different	NN	O	O
chromosomes	NN	O	O
.	NN	O	O

Production	NN	O	O
of	NN	O	O
functional	NN	O	B-protein
IL-12	NN	O	I-protein
requires	NN	O	O
the	NN	O	O
coordinated	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
genes	NN	O	O
.	NN	O	O

This	NN	O	O
presents	NN	O	O
several	NN	O	O
problems	NN	O	O
in	NN	O	O
vectors	NN	O	O
,	NN	O	O
particularly	NN	O	O
those	NN	O	O
in	NN	O	O
which	NN	O	O
additional	NN	O	O
genes	NN	O	O
,	NN	O	O
either	NN	O	O
a	NN	O	O
co-stimulatory	NN	O	B-DNA
gene	NN	O	I-DNA
or	NN	O	O
a	NN	O	O
selectable	NN	O	O
marker	NN	O	O
,	NN	O	O
are	NN	O	O
inserted	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
constructed	NN	O	O
a	NN	O	O
single	NN	O	O
cDNA	NN	O	B-DNA
that	NN	O	O
encodes	NN	O	O
a	NN	O	O
single-chain	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
called	NN	O	O
Flexi-12	NN	O	B-protein
,	NN	O	O
which	NN	O	O
retains	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
biological	NN	O	O
characteristics	NN	O	O
of	NN	O	O
recombinant	NN	O	O
IL-12	NN	O	B-protein
(	NN	O	O
rIL-12	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
monomeric	NN	O	O
polypeptide	NN	O	O
Flexi-12	NN	O	B-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
phytohemagglutinin	NN	O	B-cell_line
(	NN	O	I-cell_line
PHA	NN	O	I-cell_line
)	NN	O	I-cell_line
blasts	NN	O	I-cell_line
,	NN	O	O
induce	NN	O	O
PHA	NN	O	B-cell_line
blasts	NN	O	I-cell_line
to	NN	O	O
secrete	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
and	NN	O	O
additionally	NN	O	O
,	NN	O	O
by	NN	O	O
preincubation	NN	O	O
,	NN	O	O
enhance	NN	O	O
the	NN	O	O
killing	NN	O	O
of	NN	O	O
K562	NN	O	O
targets	NN	O	O
by	NN	O	O
PBLs	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
phenomena	NN	O	O
are	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
comparable	NN	O	O
to	NN	O	O
that	NN	O	O
seen	NN	O	O
with	NN	O	O
rIL-12	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
also	NN	O	O
shown	NN	O	O
that	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
STAT	NN	O	B-protein
4	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
unique	NN	O	O
to	NN	O	O
the	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
,	NN	O	O
occurs	NN	O	O
with	NN	O	O
Flexi-12	NN	O	B-protein
at	NN	O	O
levels	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
seen	NN	O	O
with	NN	O	O
rIL-12	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
packaged	NN	O	O
Flexi-12	NN	O	B-protein
into	NN	O	O
a	NN	O	O
recombinant	NN	O	O
adeno-associated	NN	O	O
virus	NN	O	O
(	NN	O	O
AAV	NN	O	O
)	NN	O	O
and	NN	O	O
used	NN	O	O
this	NN	O	O
vector	NN	O	O
to	NN	O	O
infect	NN	O	O
acute	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
leukemic	NN	O	I-cell_type
(	NN	O	I-cell_type
AML	NN	O	I-cell_type
)	NN	O	I-cell_type
blasts	NN	O	I-cell_type
.	NN	O	O

Infected	NN	O	O
AML	NN	O	B-cell_type
blasts	NN	O	I-cell_type
produced	NN	O	O
between	NN	O	O
2	NN	O	O
and	NN	O	O
6	NN	O	O
ng	NN	O	O
of	NN	O	O
IL-12/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
per	NN	O	O
ml	NN	O	O
per	NN	O	O
48	NN	O	O
hr	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
also	NN	O	O
confirm	NN	O	O
that	NN	O	O
AAV	NN	O	O
is	NN	O	O
an	NN	O	O
efficient	NN	O	O
delivery	NN	O	O
vehicle	NN	O	O
for	NN	O	O
cytokines	NN	O	B-protein
to	NN	O	O
leukemic	NN	O	O
cells	NN	O	O
.	NN	O	O

Direct	NN	O	O
analysis	NN	O	O
of	NN	O	O
these	NN	O	O
modified	NN	O	O
cells	NN	O	O
acting	NN	O	O
as	NN	O	O
tumor	NN	O	O
vaccines	NN	O	O
is	NN	O	O
underway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Plasma	NN	O	O
sialyltransferase	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
psychiatric	NN	O	O
disorders	NN	O	O
as	NN	O	O
a	NN	O	O
possible	NN	O	O
indicator	NN	O	O
of	NN	O	O
HPA	NN	O	O
axis	NN	O	O
function	NN	O	O
.	NN	O	O

A	NN	O	O
dysfunction	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
(	NN	O	O
HPA	NN	O	O
)	NN	O	O
axis	NN	O	O
,	NN	O	O
possibly	NN	O	O
attributed	NN	O	O
to	NN	O	O
a	NN	O	O
change	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
functionality	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
depression	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
measured	NN	O	O
both	NN	O	O
lymphocyte	NN	O	B-protein
GR	NN	O	I-protein
receptor	NN	O	O
binding	NN	O	O
parameters	NN	O	O
and	NN	O	O
plasma	NN	O	O
sialyltransferase	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
biochemical	NN	O	O
marker	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
function	NN	O	O
,	NN	O	O
in	NN	O	O
two	NN	O	O
groups	NN	O	O
of	NN	O	O
patients	NN	O	O
suffering	NN	O	O
from	NN	O	O
depression	NN	O	O
or	NN	O	O
schizophrenia	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
age-	NN	O	O
and	NN	O	O
sex-matched	NN	O	O
controls	NN	O	O
.	NN	O	O

While	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
in	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
depressed	NN	O	O
group	NN	O	O
,	NN	O	O
there	NN	O	O
were	NN	O	O
no	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
lymphocyte	NN	O	O
GR	NN	O	B-protein
binding	NN	O	O
parameters	NN	O	O
(	NN	O	O
K	NN	O	O
(	NN	O	O
m	NN	O	O
)	NN	O	O
and	NN	O	O
Bmax	NN	O	O
)	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
plasma	NN	O	B-protein
sialyltransferase	NN	O	I-protein
:	NN	O	I-protein
cortisol	NN	O	O
ratio	NN	O	O
in	NN	O	O
the	NN	O	O
depressed	NN	O	O
group	NN	O	O
suggesting	NN	O	O
an	NN	O	O
inability	NN	O	O
of	NN	O	O
the	NN	O	O
raised	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
to	NN	O	O
induce	NN	O	O
enzyme	NN	O	O
expression	NN	O	O
and	NN	O	O
this	NN	O	O
ratio	NN	O	O
may	NN	O	O
provide	NN	O	O
a	NN	O	O
useful	NN	O	O
biochemical	NN	O	O
marker	NN	O	O
of	NN	O	O
cortisol	NN	O	O
receptor	NN	O	O
function	NN	O	O
.	NN	O	O

Although	NN	O	O
there	NN	O	O
was	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
plasma	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
2	NN	O	I-protein
,	NN	O	I-protein
6	NN	O	I-protein
sialyltransferase	NN	O	I-protein
isozyme	NN	O	I-protein
in	NN	O	O
the	NN	O	O
schizophrenic	NN	O	O
group	NN	O	O
,	NN	O	O
no	NN	O	O
other	NN	O	O
changes	NN	O	O
were	NN	O	O
determined	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
total	NN	O	O
plasma	NN	O	B-protein
sialyltransferase	NN	O	I-protein
:	NN	O	O
cortisol	NN	O	O
ratio	NN	O	O
reflects	NN	O	O
HPA	NN	O	O
axis	NN	O	O
function	NN	O	O
,	NN	O	O
alterations	NN	O	O
in	NN	O	O
specific	NN	O	O
isozyme	NN	O	O
activity	NN	O	O
may	NN	O	O
also	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
other	NN	O	O
CNS	NN	O	O
disease	NN	O	O
states	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
CD95	NN	O	B-protein
(	NN	O	I-protein
Fas	NN	O	I-protein
)	NN	O	I-protein
ligand	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
events	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
mature	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
TCR	NN	O	B-protein
engagement	NN	O	O
results	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
signals	NN	O	O
that	NN	O	O
drive	NN	O	O
induction	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
clonal	NN	O	O
expansion	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
under	NN	O	O
some	NN	O	O
conditions	NN	O	O
,	NN	O	O
engagement	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
leads	NN	O	O
instead	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
TCR	NN	O	B-protein
-stimulated	NN	O	O
apoptosis	NN	O	O
requires	NN	O	O
expression	NN	O	O
of	NN	O	O
CD95	NN	O	B-protein
ligand	NN	O	I-protein
on	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
followed	NN	O	O
by	NN	O	O
an	NN	O	O
interaction	NN	O	O
between	NN	O	O
CD95	NN	O	B-protein
ligand	NN	O	I-protein
and	NN	O	O
the	NN	O	O
CD95	NN	O	B-protein
receptor	NN	O	I-protein
also	NN	O	O
expressed	NN	O	O
on	NN	O	O
this	NN	O	O
population	NN	O	O
.	NN	O	O

The	NN	O	O
experiments	NN	O	O
reported	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
were	NN	O	O
designed	NN	O	O
to	NN	O	O
address	NN	O	O
the	NN	O	O
signaling	NN	O	O
events	NN	O	O
triggered	NN	O	O
by	NN	O	O
TCR	NN	O	B-protein
engagement	NN	O	O
that	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
regulating	NN	O	O
CD95	NN	O	B-protein
ligand	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

To	NN	O	O
approach	NN	O	O
this	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
a	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
containing	NN	O	O
elements	NN	O	O
of	NN	O	O
the	NN	O	O
CD95	NN	O	B-protein
ligand	NN	O	I-protein
promoter	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
previously	NN	O	O
described	NN	O	O
mutant	NN	O	O
of	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
proximal	NN	O	O
signaling	NN	O	O
events	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
CD45	NN	O	B-protein
tyrosine	NN	O	I-protein
phosphatase	NN	O	I-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
TCR	NN	O	B-protein
-stimulated	NN	O	O
CD95	NN	O	B-protein
ligand	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
further	NN	O	O
that	NN	O	O
TCR	NN	O	B-protein
-stimulated	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
Ras	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
activation	NN	O	O
of	NN	O	O
CD95	NN	O	B-protein
ligand	NN	O	I-protein
.	NN	O	O

Next	NN	O	O
,	NN	O	O
in	NN	O	O
an	NN	O	O
effort	NN	O	O
to	NN	O	O
determine	NN	O	O
critical	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
CD95	NN	O	B-protein
ligand	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
cyclosporin	NN	O	B-DNA
A-sensitive	NN	O	I-DNA
nuclear	NN	O	I-DNA
factor-AT	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
this	NN	O	O
gene	NN	O	O
that	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
optimal	NN	O	O
CD95	NN	O	B-protein
ligand	NN	O	I-protein
reporter	NN	O	O
activity	NN	O	O
in	NN	O	O
stimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
studies	NN	O	O
begin	NN	O	O
a	NN	O	O
dissection	NN	O	O
of	NN	O	O
the	NN	O	O
biochemical	NN	O	O
events	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
expression	NN	O	O
of	NN	O	O
CD95	NN	O	B-protein
ligand	NN	O	I-protein
,	NN	O	O
a	NN	O	O
required	NN	O	O
step	NN	O	O
for	NN	O	O
TCR	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
and	NN	O	O
antioxidants	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
.	NN	O	O

This	NN	O	O
paper	NN	O	O
aims	NN	O	O
to	NN	O	O
review	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
free	NN	O	O
radical-induced	NN	O	O
tissue	NN	O	O
damage	NN	O	O
and	NN	O	O
antioxidant	NN	O	O
defence	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
that	NN	O	O
involve	NN	O	O
pathogenic	NN	O	O
processes	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
periodontal	NN	O	O
diseases	NN	O	O
.	NN	O	O

There	NN	O	O
is	NN	O	O
a	NN	O	O
clearly	NN	O	O
defined	NN	O	O
and	NN	O	O
substantial	NN	O	O
role	NN	O	O
for	NN	O	O
free	NN	O	O
radicals	NN	O	O
or	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
in	NN	O	O
periodontitis	NN	O	O
,	NN	O	O
but	NN	O	O
little	NN	O	O
research	NN	O	O
has	NN	O	O
been	NN	O	O
performed	NN	O	O
in	NN	O	O
this	NN	O	O
area	NN	O	O
.	NN	O	O

This	NN	O	O
paper	NN	O	O
reviews	NN	O	O
the	NN	O	O
considerable	NN	O	O
data	NN	O	O
available	NN	O	O
relating	NN	O	O
ROS	NN	O	O
activity	NN	O	O
and	NN	O	O
antioxidant	NN	O	O
defence	NN	O	O
to	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
and	NN	O	O
attempts	NN	O	O
to	NN	O	O
draw	NN	O	O
parallels	NN	O	O
with	NN	O	O
periodontitis	NN	O	O
,	NN	O	O
in	NN	O	O
an	NN	O	O
effort	NN	O	O
to	NN	O	O
stimulate	NN	O	O
more	NN	O	O
periodontal	NN	O	O
research	NN	O	O
in	NN	O	O
this	NN	O	O
important	NN	O	O
area	NN	O	O
.	NN	O	O

The	NN	O	O
recent	NN	O	O
discovery	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
is	NN	O	O
reviewed	NN	O	O
and	NN	O	O
several	NN	O	O
potential	NN	O	O
pathways	NN	O	O
for	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
periodontal	NN	O	O
tissue	NN	O	O
damage	NN	O	O
,	NN	O	O
mediated	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
are	NN	O	O
discussed	NN	O	O
.	NN	O	O

Emphasis	NN	O	O
is	NN	O	O
placed	NN	O	O
on	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
have	NN	O	O
been	NN	O	O
studied	NN	O	O
in	NN	O	O
periodontitis	NN	O	O
,	NN	O	O
principally	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
IL-1	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
IL-8	NN	O	B-protein
and	NN	O	O
beta-interferon	NN	O	B-protein
.	NN	O	O

The	NN	O	O
link	NN	O	O
between	NN	O	O
cellular	NN	O	O
production	NN	O	O
of	NN	O	O
such	NN	O	O
important	NN	O	O
mediators	NN	O	O
of	NN	O	O
inflammation	NN	O	O
and	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
(	NN	O	O
AO	NN	O	O
)	NN	O	O
thiols	NN	O	O
,	NN	O	O
cysteine	NN	O	O
and	NN	O	O
reduced	NN	O	O
glutathione	NN	O	O
(	NN	O	O
GSH	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
discussed	NN	O	O
and	NN	O	O
it	NN	O	O
is	NN	O	O
hypothesised	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
antagonists	NN	O	O
may	NN	O	O
offer	NN	O	O
important	NN	O	O
therapeutic	NN	O	O
benefits	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
TSC2	NN	O	B-DNA
gene	NN	O	I-DNA
:	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
complete	NN	O	O
coding	NN	O	O
sequence	NN	O	O
using	NN	O	O
the	NN	O	O
protein	NN	O	O
truncation	NN	O	O
test	NN	O	O
(	NN	O	O
PTT	NN	O	O
)	NN	O	O
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
TSC2	NN	O	B-DNA
gene	NN	O	I-DNA
on	NN	O	O
chromosome	NN	O	B-DNA
16p13.3	NN	O	I-DNA
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
approximately	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
familial	NN	O	O
tuberous	NN	O	O
sclerosis	NN	O	O
(	NN	O	O
TSC	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
has	NN	O	O
41	NN	O	B-DNA
small	NN	O	I-DNA
exons	NN	O	I-DNA
spanning	NN	O	O
45	NN	O	B-DNA
kb	NN	O	I-DNA
of	NN	O	O
genomic	NN	O	B-DNA
DNA	NN	O	I-DNA
and	NN	O	O
encoding	NN	O	O
a	NN	O	O
5.5	NN	O	B-RNA
kb	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Large	NN	O	O
germline	NN	O	O
deletions	NN	O	O
of	NN	O	O
TSC2	NN	O	B-DNA
occur	NN	O	O
in	NN	O	O
<	NN	O	O
5	NN	O	O
%	NN	O	O
of	NN	O	O
cases	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
small	NN	O	O
intragenic	NN	O	O
mutations	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

We	NN	O	O
analysed	NN	O	O
mRNA	NN	O	O
from	NN	O	O
18	NN	O	O
unrelated	NN	O	O
cases	NN	O	O
of	NN	O	O
TSC	NN	O	O
for	NN	O	O
TSC2	NN	O	B-DNA
mutations	NN	O	I-DNA
using	NN	O	O
the	NN	O	O
protein	NN	O	O
truncation	NN	O	O
test	NN	O	O
(	NN	O	O
PTT	NN	O	O
)	NN	O	O
.	NN	O	O

Three	NN	O	O
cases	NN	O	O
were	NN	O	O
predicted	NN	O	O
to	NN	O	O
be	NN	O	O
TSC2	NN	O	B-DNA
mutations	NN	O	I-DNA
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
linkage	NN	O	O
analysis	NN	O	O
or	NN	O	O
because	NN	O	O
a	NN	O	O
hamartoma	NN	O	O
from	NN	O	O
the	NN	O	O
patient	NN	O	O
showed	NN	O	O
loss	NN	O	O
of	NN	O	O
heterozygosity	NN	O	O
for	NN	O	O
16p13.3	NN	O	B-DNA
markers	NN	O	I-DNA
.	NN	O	O

Three	NN	O	O
overlapping	NN	O	O
PCR	NN	O	B-DNA
products	NN	O	I-DNA
,	NN	O	O
covering	NN	O	O
the	NN	O	O
complete	NN	O	O
coding	NN	O	O
sequence	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
,	NN	O	O
were	NN	O	O
generated	NN	O	O
from	NN	O	O
lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
translated	NN	O	O
into	NN	O	O
35S-methionine	NN	O	B-protein
labelled	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
analysed	NN	O	O
by	NN	O	O
SDS-PAGE	NN	O	O
.	NN	O	O

PCR	NN	O	B-DNA
products	NN	O	I-DNA
showing	NN	O	O
PTT	NN	O	O
shifts	NN	O	O
were	NN	O	O
directly	NN	O	O
sequenced	NN	O	O
,	NN	O	O
and	NN	O	O
mutations	NN	O	O
confirmed	NN	O	O
by	NN	O	O
restriction	NN	O	O
enzyme	NN	O	O
digestion	NN	O	O
where	NN	O	O
possible	NN	O	O
.	NN	O	O

Six	NN	O	O
PTT	NN	O	O
shifts	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Five	NN	O	O
of	NN	O	O
these	NN	O	O
were	NN	O	O
caused	NN	O	O
by	NN	O	O
mutations	NN	O	O
predicted	NN	O	O
to	NN	O	O
produce	NN	O	O
a	NN	O	O
truncated	NN	O	B-protein
protein	NN	O	I-protein
:	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
a	NN	O	O
sporadic	NN	O	O
case	NN	O	O
showed	NN	O	O
a	NN	O	O
32	NN	O	B-DNA
bp	NN	O	I-DNA
deletion	NN	O	I-DNA
in	NN	O	O
exon	NN	O	B-RNA
11	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
mutant	NN	O	B-RNA
mRNA	NN	O	I-RNA
without	NN	O	O
exon	NN	O	B-RNA
11	NN	O	I-RNA
was	NN	O	O
produced	NN	O	O
;	NN	O	O
the	NN	O	O
normal	NN	O	O
exon	NN	O	O
10	NN	O	O
was	NN	O	O
also	NN	O	O
spliced	NN	O	O
out	NN	O	O
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
a	NN	O	O
sporadic	NN	O	O
case	NN	O	O
had	NN	O	O
a	NN	O	O
1	NN	O	B-DNA
bp	NN	O	I-DNA
deletion	NN	O	I-DNA
in	NN	O	O
exon	NN	O	B-RNA
12	NN	O	I-RNA
(	NN	O	O
1634delT	NN	O	B-RNA
)	NN	O	O
;	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
a	NN	O	O
TSC2-linked	NN	O	B-RNA
mother	NN	O	I-RNA
and	NN	O	I-RNA
daughter	NN	O	I-RNA
pair	NN	O	I-RNA
had	NN	O	O
a	NN	O	O
G	NN	O	O
--	NN	O	O
>	NN	O	O
T	NN	O	O
transversion	NN	O	O
in	NN	O	O
exon	NN	O	B-RNA
23	NN	O	I-RNA
(	NN	O	O
G2715T	NN	O	B-RNA
)	NN	O	O
introducing	NN	O	O
a	NN	O	O
cryptic	NN	O	O
splice	NN	O	O
site	NN	O	O
causing	NN	O	O
a	NN	O	O
29	NN	O	O
bp	NN	O	O
truncation	NN	O	O
of	NN	O	O
mRNA	NN	O	O
from	NN	O	O
exon	NN	O	B-RNA
23	NN	O	I-RNA
;	NN	O	O
(	NN	O	O
iv	NN	O	O
)	NN	O	O
a	NN	O	O
sporadic	NN	O	O
case	NN	O	O
showed	NN	O	O
a	NN	O	O
2	NN	O	B-DNA
bp	NN	O	I-DNA
deletion	NN	O	I-DNA
in	NN	O	O
exon	NN	O	B-RNA
36	NN	O	I-RNA
;	NN	O	O
(	NN	O	O
v	NN	O	O
)	NN	O	O
a	NN	O	O
sporadic	NN	O	O
case	NN	O	O
showed	NN	O	O
a	NN	O	O
1	NN	O	B-DNA
bp	NN	O	I-DNA
insertion	NN	O	I-DNA
disrupting	NN	O	O
the	NN	O	O
donor	NN	O	O
splice	NN	O	O
site	NN	O	O
of	NN	O	O
exon	NN	O	B-RNA
37	NN	O	I-RNA
(	NN	O	O
5007+2insA	NN	O	B-RNA
)	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
an	NN	O	O
upstream	NN	O	B-RNA
exonic	NN	O	I-RNA
cryptic	NN	O	I-RNA
splice	NN	O	I-RNA
site	NN	O	I-RNA
to	NN	O	O
cause	NN	O	O
a	NN	O	O
29	NN	O	B-RNA
bp	NN	O	I-RNA
truncation	NN	O	I-RNA
of	NN	O	O
mRNA	NN	O	B-RNA
from	NN	O	O
exon	NN	O	B-RNA
37	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
one	NN	O	O
case	NN	O	O
,	NN	O	O
the	NN	O	O
PTT	NN	O	O
shift	NN	O	O
was	NN	O	O
explained	NN	O	O
by	NN	O	O
in-frame	NN	O	O
splicing	NN	O	O
out	NN	O	O
of	NN	O	O
exon	NN	O	B-RNA
10	NN	O	I-RNA
,	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
normal	NN	O	B-DNA
exon	NN	O	I-DNA
10	NN	O	I-DNA
genomic	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Alternative	NN	O	O
splicing	NN	O	O
of	NN	O	O
exon	NN	O	B-RNA
10	NN	O	I-RNA
of	NN	O	O
the	NN	O	O
TSC2	NN	O	B-DNA
gene	NN	O	I-DNA
may	NN	O	O
be	NN	O	O
a	NN	O	O
normal	NN	O	O
variant	NN	O	O
.	NN	O	O

Three	NN	O	O
3rd	NN	O	O
base	NN	O	O
substitution	NN	O	O
polymorphisms	NN	O	O
were	NN	O	O
also	NN	O	O
detected	NN	O	O
during	NN	O	O
direct	NN	O	O
sequencing	NN	O	O
of	NN	O	O
PCR	NN	O	B-DNA
products	NN	O	I-DNA
.	NN	O	O

Confirmed	NN	O	O
mutations	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
28	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
families	NN	O	O
studied	NN	O	O
and	NN	O	O
on	NN	O	O
the	NN	O	O
assumption	NN	O	O
that	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
sporadic	NN	O	O
cases	NN	O	O
should	NN	O	O
have	NN	O	O
TSC2	NN	O	B-DNA
mutations	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
crude	NN	O	O
estimate	NN	O	O
of	NN	O	O
the	NN	O	O
detection	NN	O	O
rate	NN	O	O
would	NN	O	O
be	NN	O	O
60	NN	O	O
%	NN	O	O
.	NN	O	O

This	NN	O	O
compares	NN	O	O
favourably	NN	O	O
with	NN	O	O
other	NN	O	O
screening	NN	O	O
methods	NN	O	O
used	NN	O	O
for	NN	O	O
TSC2	NN	O	B-DNA
,	NN	O	O
notably	NN	O	O
SSCP	NN	O	O
,	NN	O	O
and	NN	O	O
since	NN	O	O
PTT	NN	O	O
involves	NN	O	O
much	NN	O	O
less	NN	O	O
work	NN	O	O
it	NN	O	O
may	NN	O	O
be	NN	O	O
the	NN	O	O
method	NN	O	O
of	NN	O	O
choice	NN	O	O
.	NN	O	O

-DOCSTART-	O

alpha-Tocopheryl	NN	O	O
succinate	NN	O	O
inhibits	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
suppressing	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
mobilization	NN	O	O
.	NN	O	O

The	NN	O	O
adherence	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
to	NN	O	O
activated	NN	O	O
endothelium	NN	O	O
is	NN	O	O
an	NN	O	O
early	NN	O	O
event	NN	O	O
in	NN	O	O
atherogenesis	NN	O	O
.	NN	O	O

Because	NN	O	O
antioxidants	NN	O	O
have	NN	O	O
been	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
of	NN	O	O
antiatherosclerotic	NN	O	O
potential	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
alpha-tocopherol	NN	O	O
(	NN	O	O
TCP	NN	O	O
)	NN	O	O
and	NN	O	O
its	NN	O	O
acetate	NN	O	O
and	NN	O	O
succinate	NN	O	O
esters	NN	O	O
on	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
cytokine-stimulated	NN	O	B-cell_line
human	NN	O	I-cell_line
umbilical	NN	O	I-cell_line
vein	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
HUVEC	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

Endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
treated	NN	O	O
with	NN	O	O
TCP	NN	O	O
,	NN	O	O
alpha-tocopherol	NN	O	O
acetate	NN	O	O
(	NN	O	O
TCP	NN	O	O
acetate	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
alpha-tocopheryl	NN	O	O
succinate	NN	O	O
(	NN	O	O
TCP	NN	O	O
succinate	NN	O	O
)	NN	O	O
before	NN	O	O
stimulation	NN	O	O
with	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
;	NN	O	O
10	NN	O	O
U/ml	NN	O	O
,	NN	O	O
6	NN	O	O
h	NN	O	O
)	NN	O	O
or	NN	O	O
interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
(	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
;	NN	O	O
10	NN	O	O
U/ml	NN	O	O
,	NN	O	O
6	NN	O	O
h	NN	O	O
)	NN	O	O
.	NN	O	O

Cytokine-stimulated	NN	O	O
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
vascular	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
VCAM-1	NN	O	B-protein
,	NN	O	O
CD106	NN	O	B-protein
)	NN	O	O
and	NN	O	O
E-selectin	NN	O	B-protein
(	NN	O	O
ELAM-1	NN	O	B-protein
,	NN	O	O
CD62E	NN	O	B-protein
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
CD54	NN	O	B-protein
)	NN	O	O
,	NN	O	O
was	NN	O	O
time-	NN	O	O
and	NN	O	O
dose-dependently	NN	O	O
inhibited	NN	O	O
by	NN	O	O
TCP	NN	O	O
succinate	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
TCP	NN	O	O
or	NN	O	O
TCP	NN	O	O
acetate	NN	O	O
.	NN	O	O

TCP	NN	O	O
succinate	NN	O	O
(	NN	O	O
200	NN	O	O
microM	NN	O	O
,	NN	O	O
24	NN	O	O
h	NN	O	O
)	NN	O	O
reduced	NN	O	O
TNF-induced	NN	O	B-protein
VCAM-1	NN	O	I-protein
and	NN	O	O
E-selectin	NN	O	B-protein
expression	NN	O	O
from	NN	O	O
a	NN	O	O
specific	NN	O	O
mean	NN	O	O
fluorescence	NN	O	O
intensity	NN	O	O
of	NN	O	O
151	NN	O	O
+/-	NN	O	O
28	NN	O	O
to	NN	O	O
12	NN	O	O
+/-	NN	O	O
4	NN	O	O
channels	NN	O	O
and	NN	O	O
from	NN	O	O
225	NN	O	O
+/-	NN	O	O
38	NN	O	O
to	NN	O	O
79	NN	O	O
+/-	NN	O	O
21	NN	O	O
channels	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Succinate	NN	O	O
alone	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

Decreased	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
expression	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
reduction	NN	O	O
of	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
.	NN	O	O

TCP	NN	O	O
succinate	NN	O	O
(	NN	O	O
20	NN	O	O
microM	NN	O	O
,	NN	O	O
72	NN	O	O
h	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
TCP	NN	O	O
(	NN	O	O
200	NN	O	O
microM	NN	O	O
,	NN	O	O
72	NN	O	O
h	NN	O	O
)	NN	O	O
,	NN	O	O
reduced	NN	O	O
U-937	NN	O	B-cell_line
cell	NN	O	I-cell_line
adhesion	NN	O	O
to	NN	O	O
TNF-alpha-stimulated	NN	O	B-cell_line
(	NN	O	I-cell_line
10	NN	O	I-cell_line
U/ml	NN	O	I-cell_line
,	NN	O	I-cell_line
6	NN	O	I-cell_line
h	NN	O	I-cell_line
)	NN	O	I-cell_line
HUVEC	NN	O	I-cell_line
by	NN	O	O
30	NN	O	O
%	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.025	NN	O	O
)	NN	O	O
and	NN	O	O
to	NN	O	O
IL-1	NN	O	B-cell_line
beta-stimulated	NN	O	I-cell_line
HUVEC	NN	O	I-cell_line
by	NN	O	O
56	NN	O	O
%	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.010	NN	O	O
)	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility-shift	NN	O	O
assays	NN	O	O
of	NN	O	O
HUVEC	NN	O	B-cell_line
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
revealed	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
TNF-alpha-stimulated	NN	O	B-protein
nuclear	NN	O	I-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
activation	NN	O	O
after	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
HUVEC	NN	O	B-cell_line
with	NN	O	O
TCP	NN	O	O
succinate	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
TCP	NN	O	O
,	NN	O	O
TCP	NN	O	O
acetate	NN	O	O
,	NN	O	O
or	NN	O	O
succinate	NN	O	O
alone	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
vitamin	NN	O	O
E	NN	O	O
derivative	NN	O	O
TCP	NN	O	O
succinate	NN	O	O
prevents	NN	O	O
monocytic	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
to	NN	O	O
cytokine-stimulated	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
further	NN	O	O
emphasizing	NN	O	O
the	NN	O	O
antiatherosclerotic	NN	O	O
potential	NN	O	O
of	NN	O	O
lipid	NN	O	O
soluble	NN	O	O
antioxidants	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
homolog	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-DNA
extra	NN	O	I-DNA
sex	NN	O	I-DNA
combs	NN	O	I-DNA
gene	NN	O	I-DNA
displays	NN	O	O
transcriptional	NN	O	B-protein
repressor	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

The	NN	O	O
heterogeneous	NN	O	B-protein
nuclear	NN	O	I-protein
ribonucleoprotein	NN	O	I-protein
K	NN	O	I-protein
protein	NN	O	I-protein
represents	NN	O	O
a	NN	O	O
novel	NN	O	O
class	NN	O	O
of	NN	O	O
proteins	NN	O	O
that	NN	O	O
may	NN	O	O
act	NN	O	O
as	NN	O	O
docking	NN	O	O
platforms	NN	O	O
that	NN	O	O
orchestrate	NN	O	O
cross-talk	NN	O	O
among	NN	O	O
molecules	NN	O	O
involved	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
and	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
fragment	NN	O	O
of	NN	O	O
K	NN	O	B-protein
protein	NN	O	I-protein
as	NN	O	O
bait	NN	O	O
in	NN	O	O
the	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
screen	NN	O	O
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
that	NN	O	O
encodes	NN	O	O
a	NN	O	O
protein	NN	O	O
whose	NN	O	O
primary	NN	O	O
structure	NN	O	O
has	NN	O	O
extensive	NN	O	O
similarity	NN	O	O
to	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-protein
melanogaster	NN	O	I-protein
extra	NN	O	I-protein
sex	NN	O	I-protein
combs	NN	O	I-protein
(	NN	O	I-protein
esc	NN	O	I-protein
)	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
,	NN	O	O
Esc	NN	O	B-protein
,	NN	O	O
a	NN	O	O
putative	NN	O	O
silencer	NN	O	O
of	NN	O	O
homeotic	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
cDNA	NN	O	B-DNA
that	NN	O	O
we	NN	O	O
isolated	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
cDNA	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
recently	NN	O	O
positionally	NN	O	O
cloned	NN	O	O
mouse	NN	O	B-DNA
embryonic	NN	O	I-DNA
ectoderm	NN	O	I-DNA
development	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
eed	NN	O	O
.	NN	O	O

Like	NN	O	O
Esc	NN	O	B-protein
,	NN	O	O
Eed	NN	O	B-protein
contains	NN	O	O
six	NN	O	O
WD-40	NN	O	B-protein
repeats	NN	O	I-protein
in	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
half	NN	O	I-protein
of	NN	O	O
the	NN	O	O
protein	NN	O	O
and	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
repress	NN	O	O
homeotic	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
mouse	NN	O	O
embryogenesis	NN	O	O
.	NN	O	O

Eed	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
K	NN	O	B-protein
protein	NN	O	I-protein
through	NN	O	O
a	NN	O	O
domain	NN	O	O
in	NN	O	O
its	NN	O	O
N	NN	O	B-protein
terminus	NN	O	I-protein
,	NN	O	O
but	NN	O	O
interestingly	NN	O	O
,	NN	O	O
this	NN	O	O
domain	NN	O	O
is	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
Drosophila	NN	O	O
Esc	NN	O	B-protein
.	NN	O	O

Gal4-Eed	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
represses	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	O
gene	NN	O	O
driven	NN	O	O
by	NN	O	O
a	NN	O	O
promoter	NN	O	O
that	NN	O	O
contains	NN	O	O
Gal4-binding	NN	O	B-DNA
DNA	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Eed	NN	O	B-protein
also	NN	O	O
represses	NN	O	O
transcription	NN	O	O
when	NN	O	O
recruited	NN	O	O
to	NN	O	O
a	NN	O	O
target	NN	O	O
promoter	NN	O	O
by	NN	O	O
Gal4-K	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Point	NN	O	O
mutations	NN	O	O
within	NN	O	O
the	NN	O	O
eed	NN	O	O
gene	NN	O	O
that	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
severe	NN	O	O
embryonic	NN	O	O
development	NN	O	O
abnormalities	NN	O	O
abolished	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
repressor	NN	O	I-protein
activity	NN	O	O
of	NN	O	O
Eed	NN	O	B-protein
.	NN	O	O

Results	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
suggest	NN	O	O
that	NN	O	O
Eed	NN	O	B-protein
-restricted	NN	O	O
homeotic	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
embryogenesis	NN	O	O
reflects	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
Eed	NN	O	B-protein
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
repressor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Eed	NN	O	B-protein
-mediated	NN	O	O
transcriptional	NN	O	O
effects	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
reflect	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
Eed	NN	O	B-protein
with	NN	O	O
multiple	NN	O	O
molecular	NN	O	O
partners	NN	O	O
,	NN	O	O
including	NN	O	O
K	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Interferons	NN	O	B-protein
up-regulate	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	O
STAT2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IRF	NN	O	B-protein
family	NN	O	O
transcription	NN	O	O
factor	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

IFN	NN	O	B-protein
signaling	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
binding	NN	O	O
of	NN	O	O
IFNs	NN	O	B-protein
to	NN	O	O
their	NN	O	O
receptors	NN	O	O
and	NN	O	O
subsequent	NN	O	O
activation	NN	O	O
of	NN	O	O
Janus	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
(	NN	O	O
JAK	NN	O	B-protein
)	NN	O	O
-	NN	O	O
STAT	NN	O	B-protein
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
IFN-alpha	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
the	NN	O	O
assembly	NN	O	O
of	NN	O	O
IFN-stimulated	NN	O	B-protein
gene	NN	O	I-protein
factor	NN	O	I-protein
3	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complex	NN	O	I-protein
formed	NN	O	O
by	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	O
STAT2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p48	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

IFN-gamma	NN	O	B-protein
signaling	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
homodimeric	NN	O	B-protein
STAT1	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
these	NN	O	O
signaling	NN	O	O
molecules	NN	O	O
are	NN	O	O
expressed	NN	O	O
constitutively	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
also	NN	O	O
evidence	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
by	NN	O	O
IFNs	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
characterized	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
and	NN	O	I-protein
IFN	NN	O	I-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
IRF	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
IFN-alpha	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	O
stimulation	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
IFN-alpha	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
rapidly	NN	O	O
and	NN	O	O
efficiently	NN	O	O
enhanced	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	O
STAT2	NN	O	B-protein
,	NN	O	O
p48	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IRF-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

IFN-gamma	NN	O	B-protein
induced	NN	O	O
IRF-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
more	NN	O	O
strongly	NN	O	O
than	NN	O	O
IFN-alpha	NN	O	B-protein
.	NN	O	O

Stimulation	NN	O	O
experiments	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
cycloheximide	NN	O	O
,	NN	O	O
suggested	NN	O	O
that	NN	O	O
these	NN	O	O
genes	NN	O	O
were	NN	O	O
activated	NN	O	O
directly	NN	O	O
by	NN	O	O
IFNs	NN	O	B-protein
.	NN	O	O

IRF-2	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
apparently	NN	O	O
only	NN	O	O
weakly	NN	O	O
responsive	NN	O	O
to	NN	O	O
IFNs	NN	O	B-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

When	NN	O	O
macrophages	NN	O	B-cell_type
were	NN	O	O
pretreated	NN	O	O
with	NN	O	O
low	NN	O	O
doses	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
then	NN	O	O
stimulated	NN	O	O
with	NN	O	O
IFN-alpha	NN	O	B-protein
,	NN	O	O
clearly	NN	O	O
enhanced	NN	O	O
formation	NN	O	O
of	NN	O	O
specific	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
complexes	NN	O	O
was	NN	O	O
detected	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
higher	NN	O	O
intracellular	NN	O	O
levels	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
,	NN	O	O
STAT2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p48	NN	O	B-protein
protein	NN	O	I-protein
may	NN	O	O
result	NN	O	O
in	NN	O	O
enhanced	NN	O	O
signal	NN	O	O
transduction	NN	O	O
for	NN	O	O
cytokines	NN	O	B-protein
utilizing	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
regulation	NN	O	O
.	NN	O	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
for	NN	O	O
maximal	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
adult	NN	O	O
respiratory	NN	O	O
distress	NN	O	O
syndrome	NN	O	O
(	NN	O	O
ARDS	NN	O	O
)	NN	O	O
and	NN	O	O
sepsis	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Activation	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
are	NN	O	O
tightly	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
inhibitory	NN	O	B-protein
proteins	NN	O	I-protein
(	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
that	NN	O	O
sequester	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
immune/inflammatory	NN	O	B-cell_type
effector	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
involves	NN	O	O
signaled	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
ubiquitination	NN	O	O
,	NN	O	O
and	NN	O	O
proteolysis	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Liberated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
migrates	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
where	NN	O	O
it	NN	O	O
binds	NN	O	O
to	NN	O	O
specific	NN	O	B-DNA
promoter	NN	O	I-DNA
sites	NN	O	I-DNA
and	NN	O	O
activates	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
initiates	NN	O	O
both	NN	O	O
extracellular	NN	O	O
and	NN	O	O
intracellular	NN	O	O
regulatory	NN	O	O
events	NN	O	O
that	NN	O	O
result	NN	O	O
in	NN	O	O
autoregulation	NN	O	O
of	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
cascade	NN	O	O
through	NN	O	O
modulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
has	NN	O	O
been	NN	O	O
linked	NN	O	O
to	NN	O	O
ARDS	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
critical	NN	O	O
proximal	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
neutrophilic	NN	O	O
inflammation	NN	O	O
in	NN	O	O
animal	NN	O	O
models	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
can	NN	O	O
be	NN	O	O
inhibited	NN	O	O
in	NN	O	O
vivo	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
antioxidants	NN	O	O
,	NN	O	O
corticosteroids	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
endotoxin	NN	O	B-protein
tolerance	NN	O	O
.	NN	O	O

Identification	NN	O	O
of	NN	O	O
more	NN	O	O
specific	NN	O	O
and	NN	O	O
efficacious	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
might	NN	O	O
prove	NN	O	O
beneficial	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
cytokine-mediated	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

Chimeric	NN	O	B-protein
oncoprotein	NN	O	I-protein
E2a-Pbx1	NN	O	B-protein
induces	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
a	NN	O	O
p53	NN	O	B-protein
-independent	NN	O	O
mechanism	NN	O	O
that	NN	O	O
is	NN	O	O
suppressed	NN	O	O
by	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
chimeric	NN	O	B-protein
oncoprotein	NN	O	I-protein
E2a-Pbx1	NN	O	B-protein
results	NN	O	O
from	NN	O	O
fusion	NN	O	O
of	NN	O	O
the	NN	O	O
E2A	NN	O	B-DNA
and	NN	O	I-DNA
PBX1	NN	O	I-DNA
genes	NN	O	I-DNA
following	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
1	NN	O	I-DNA
;	NN	O	I-DNA
19	NN	O	I-DNA
)	NN	O	I-DNA
chromosomal	NN	O	I-DNA
translocations	NN	O	I-DNA
in	NN	O	O
B	NN	O	O
cell	NN	O	O
precursor	NN	O	O
acute	NN	O	O
leukemias	NN	O	O
.	NN	O	O

Experimentally	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
do	NN	O	O
not	NN	O	O
tolerate	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
E2a-Pbx1	NN	O	B-protein
which	NN	O	O
contrasts	NN	O	O
with	NN	O	O
transformation	NN	O	O
of	NN	O	O
several	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
following	NN	O	O
its	NN	O	O
stable	NN	O	O
expression	NN	O	O
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
investigate	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
E2a-Pbx1	NN	O	B-protein
on	NN	O	O
the	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
conditionally	NN	O	O
expressed	NN	O	O
E2a-Pbx1	NN	O	B-protein
under	NN	O	O
control	NN	O	O
of	NN	O	O
a	NN	O	O
metal	NN	O	O
response	NN	O	O
element	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_line
precursor	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
E2a-Pbx1	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
cell	NN	O	O
death	NN	O	O
with	NN	O	O
the	NN	O	O
morphologic	NN	O	O
and	NN	O	O
molecular	NN	O	O
features	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

A	NN	O	O
structure-function	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
not	NN	O	O
a	NN	O	O
dominant-negative	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
E2a	NN	O	B-protein
moiety	NN	O	I-protein
but	NN	O	O
,	NN	O	O
rather	NN	O	O
,	NN	O	O
required	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
homeodomain	NN	O	I-protein
of	NN	O	O
Pbx1	NN	O	B-protein
.	NN	O	O

E2a-	NN	O	O
Pbx1	NN	O	B-protein
-induced	NN	O	O
apoptosis	NN	O	O
proceeded	NN	O	O
through	NN	O	O
a	NN	O	O
BCL2	NN	O	B-protein
-responsive	NN	O	O
checkpoint	NN	O	O
eventuating	NN	O	O
in	NN	O	O
PARP	NN	O	B-protein
inactivation	NN	O	O
but	NN	O	O
did	NN	O	O
require	NN	O	O
p53	NN	O	B-protein
.	NN	O	O

Constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
E2a-Pbx1	NN	O	B-protein
did	NN	O	O
not	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
or	NN	O	O
continued	NN	O	O
cycling	NN	O	O
of	NN	O	O
Rat-1	NN	O	B-cell_line
fibroblasts	NN	O	I-cell_line
in	NN	O	O
low	NN	O	O
serum	NN	O	O
conditions	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
E2a-Pbx1	NN	O	B-protein
initiates	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
precursers	NN	O	I-cell_type
by	NN	O	O
a	NN	O	O
mechanism	NN	O	O
that	NN	O	O
requires	NN	O	O
its	NN	O	O
chimeric	NN	O	O
transcriptional	NN	O	O
properties	NN	O	O
,	NN	O	O
but	NN	O	O
,	NN	O	O
unlike	NN	O	O
other	NN	O	O
nuclear	NN	O	B-protein
oncoproteins	NN	O	I-protein
,	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
p53	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

CCAAT/enhancer	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
epsilon	NN	O	I-protein
is	NN	O	O
preferentially	NN	O	O
up-regulated	NN	O	O
during	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
and	NN	O	O
its	NN	O	O
functional	NN	O	O
versatility	NN	O	O
is	NN	O	O
determined	NN	O	O
by	NN	O	O
alternative	NN	O	O
use	NN	O	O
of	NN	O	O
promoters	NN	O	B-DNA
and	NN	O	O
differential	NN	O	O
splicing	NN	O	O
.	NN	O	O

CCAAT/enhancer	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
C/EBP	NN	O	I-protein
)	NN	O	I-protein
epsilon	NN	O	I-protein
is	NN	O	O
a	NN	O	O
recently	NN	O	O
cloned	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
C/EBP	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
exclusively	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
hematopoietic	NN	O	O
origin	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
C/EBPepsilon	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
transcribed	NN	O	O
by	NN	O	O
two	NN	O	O
alternative	NN	O	O
promoters	NN	O	B-DNA
,	NN	O	O
Palpha	NN	O	B-DNA
and	NN	O	O
Pbeta	NN	O	B-DNA
.	NN	O	O

A	NN	O	O
combination	NN	O	O
of	NN	O	O
differential	NN	O	O
splicing	NN	O	O
and	NN	O	O
alternative	NN	O	O
use	NN	O	O
of	NN	O	O
promoters	NN	O	B-DNA
generates	NN	O	O
four	NN	O	O
mRNA	NN	O	B-RNA
isoforms	NN	O	I-RNA
,	NN	O	O
of	NN	O	O
2.6	NN	O	O
kb	NN	O	O
and	NN	O	O
1.3-1.5	NN	O	O
kb	NN	O	O
in	NN	O	O
size	NN	O	O
.	NN	O	O

These	NN	O	O
transcripts	NN	O	O
can	NN	O	O
encode	NN	O	O
three	NN	O	O
proteins	NN	O	O
of	NN	O	O
calculated	NN	O	O
molecular	NN	O	O
mass	NN	O	O
32.2	NN	O	B-RNA
kDa	NN	O	I-RNA
,	NN	O	O
27.8	NN	O	B-RNA
kDa	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
14.3	NN	O	B-RNA
kDa	NN	O	I-RNA
.	NN	O	O

Accordingly	NN	O	O
,	NN	O	O
Western	NN	O	O
blots	NN	O	O
with	NN	O	O
antibodies	NN	O	B-protein
specific	NN	O	O
for	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
domain	NN	O	O
,	NN	O	O
that	NN	O	O
is	NN	O	O
common	NN	O	O
to	NN	O	O
all	NN	O	O
forms	NN	O	O
,	NN	O	O
identify	NN	O	O
multiple	NN	O	O
proteins	NN	O	O
.	NN	O	O

C/EBPepsilon	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
greatly	NN	O	O
induced	NN	O	O
during	NN	O	O
in	NN	O	O
vitro	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
primary	NN	O	I-cell_type
CD34	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
treatment	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
promyelocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
for	NN	O	O
24	NN	O	O
hr	NN	O	O
induced	NN	O	O
C/EBPepsilon	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
by	NN	O	O
4-fold	NN	O	O
,	NN	O	O
while	NN	O	O
prolonged	NN	O	O
treatment	NN	O	O
gradually	NN	O	O
reduced	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
to	NN	O	O
pretreatment	NN	O	O
levels	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
with	NN	O	O
expression	NN	O	O
vectors	NN	O	O
for	NN	O	O
two	NN	O	O
of	NN	O	O
the	NN	O	O
isoforms	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
32.2-kDa	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
granulocyte	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
receptor	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
while	NN	O	O
the	NN	O	O
14.3-kDa	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
not	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
C/EBPepsilon	NN	O	B-protein
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
a	NN	O	O
complex	NN	O	O
fashion	NN	O	O
and	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
TPA	NN	O	B-protein
,	NN	O	O
bryostatin	NN	O	B-protein
1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
on	NN	O	O
PO-B	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
AP-2	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
during	NN	O	O
HL-60	NN	O	O
differentiation	NN	O	O
.	NN	O	O

PO-B	NN	O	B-protein
was	NN	O	O
originally	NN	O	O
characterized	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
pro-opiomelanocortin	NN	O	B-DNA
(	NN	O	I-DNA
POMC	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
;	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
become	NN	O	O
increasingly	NN	O	O
clear	NN	O	O
that	NN	O	O
this	NN	O	O
protein	NN	O	O
may	NN	O	O
be	NN	O	O
active	NN	O	O
in	NN	O	O
tissues	NN	O	O
outside	NN	O	O
the	NN	O	O
pituitary	NN	O	O
,	NN	O	O
since	NN	O	O
it	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
diverse	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
including	NN	O	O
differentiated	NN	O	O
HL-60	NN	O	B-cell_type
promyelocytic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
previously	NN	O	O
showed	NN	O	O
that	NN	O	O
PO-B	NN	O	B-protein
DNA-binding	NN	O	O
is	NN	O	O
progressively	NN	O	O
induced	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
promyelomonocytic	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
the	NN	O	O
macrophage-like	NN	O	B-cell_type
lineage	NN	O	I-cell_type
(	NN	O	O
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
that	NN	O	O
PO-B	NN	O	B-protein
DNA-binding	NN	O	O
in	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
similarly	NN	O	O
induced	NN	O	O
during	NN	O	O
differentiation	NN	O	O
to	NN	O	O
the	NN	O	O
granulocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
(	NN	O	O
with	NN	O	O
either	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
or	NN	O	O
dimethylsulfoxide	NN	O	O
)	NN	O	O
.	NN	O	O

Either	NN	O	O
a	NN	O	O
genetic	NN	O	O
or	NN	O	O
pharmacologic	NN	O	O
blockade	NN	O	O
of	NN	O	O
HL-60	NN	O	O
differentiation	NN	O	O
prohibited	NN	O	O
these	NN	O	O
inductive	NN	O	O
effects	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
have	NN	O	O
prompted	NN	O	O
our	NN	O	O
interest	NN	O	O
in	NN	O	O
the	NN	O	O
dynamics	NN	O	O
of	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
changes	NN	O	O
during	NN	O	O
HL-60	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Of	NN	O	O
these	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
another	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
also	NN	O	O
robustly	NN	O	O
induced	NN	O	O
at	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
level	NN	O	O
during	NN	O	O
macrophage-like	NN	O	O
HL-60	NN	O	O
differentiation	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
during	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-2	NN	O	B-protein
was	NN	O	O
slightly	NN	O	O
reduced	NN	O	O
by	NN	O	O
TPA	NN	O	B-protein
-induced	NN	O	O
HL-60	NN	O	O
differentiation	NN	O	O
but	NN	O	O
unchanged	NN	O	O
during	NN	O	O
granulocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

From	NN	O	O
these	NN	O	O
data	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
PO-B	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
is	NN	O	O
a	NN	O	O
general	NN	O	O
marker	NN	O	O
of	NN	O	O
HL-60	NN	O	O
myelomonocytic	NN	O	O
differentiation	NN	O	O
,	NN	O	O
but	NN	O	O
that	NN	O	O
qualitative	NN	O	O
aspects	NN	O	O
of	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
additional	NN	O	O
distinct	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
lineage-specific	NN	O	O
determinants	NN	O	O
of	NN	O	O
cell	NN	O	O
fate	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
genetic	NN	O	O
system	NN	O	O
to	NN	O	O
isolate	NN	O	O
a	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
effector	NN	O	I-protein
on	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
Oct-2	NN	O	B-protein
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
revealed	NN	O	O
that	NN	O	O
interactions	NN	O	O
between	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
eukaryotic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
isolate	NN	O	O
cDNA	NN	O	O
clones	NN	O	O
that	NN	O	O
dominantly	NN	O	O
inhibit	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
chosen	NN	O	O
as	NN	O	O
a	NN	O	O
representative	NN	O	O
factor	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
novel	NN	O	O
screening	NN	O	O
system	NN	O	O
.	NN	O	O

This	NN	O	O
employs	NN	O	O
an	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
tester	NN	O	O
strain	NN	O	O
carrying	NN	O	O
a	NN	O	O
modified	NN	O	B-DNA
lac	NN	O	I-DNA
operon	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
with	NN	O	O
the	NN	O	O
lac	NN	O	O
operator	NN	O	O
sequence	NN	O	O
replaced	NN	O	O
by	NN	O	O
an	NN	O	O
octamer	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Oct-2	NN	O	B-protein
expressed	NN	O	O
in	NN	O	O
this	NN	O	O
tester	NN	O	O
strain	NN	O	O
represses	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
reporter	NN	O	O
gene	NN	O	O
and	NN	O	O
changes	NN	O	O
the	NN	O	O
phenotype	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
from	NN	O	O
Lac+to	NN	O	O
Lac-	NN	O	O
.	NN	O	O

Introduction	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	O
expression	NN	O	O
library	NN	O	O
prepared	NN	O	O
from	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
into	NN	O	O
the	NN	O	O
Oct-2	NN	O	B-protein
-harboring	NN	O	O
tester	NN	O	O
strain	NN	O	O
allowed	NN	O	O
selection	NN	O	O
of	NN	O	O
three	NN	O	O
Lac+clones	NN	O	O
out	NN	O	O
of	NN	O	O
1	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
5	NN	O	O
)	NN	O	O
transformants	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
them	NN	O	O
,	NN	O	O
hT86	NN	O	O
,	NN	O	O
encoding	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
derepress	NN	O	O
beta-galactosidase	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
the	NN	O	O
Oct-2	NN	O	B-protein
-harboring	NN	O	O
tester	NN	O	O
strain	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

In	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
hT86	NN	O	O
attenuated	NN	O	O
the	NN	O	O
intensity	NN	O	O
of	NN	O	O
the	NN	O	O
retarded	NN	O	O
band	NN	O	O
composed	NN	O	O
of	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
probe	NN	O	I-DNA
and	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
Oct-2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
strategy	NN	O	O
described	NN	O	O
here	NN	O	O
provides	NN	O	O
a	NN	O	O
new	NN	O	O
approach	NN	O	O
for	NN	O	O
studying	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
that	NN	O	O
govern	NN	O	O
the	NN	O	O
complex	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tap	NN	O	B-protein
:	NN	O	O
a	NN	O	O
novel	NN	O	O
cellular	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
interacts	NN	O	O
with	NN	O	O
tip	NN	O	O
of	NN	O	O
herpesvirus	NN	O	O
saimiri	NN	O	O
and	NN	O	O
induces	NN	O	O
lymphocyte	NN	O	B-cell_type
aggregation	NN	O	O
.	NN	O	O

Tip	NN	O	O
of	NN	O	O
herpesvirus	NN	O	O
saimiri	NN	O	O
associates	NN	O	O
with	NN	O	O
Lck	NN	O	B-protein
and	NN	O	O
down-regulates	NN	O	O
Lck	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	O
cellular	NN	O	O
Tip-associated	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
Tap	NN	O	B-protein
)	NN	O	O
by	NN	O	O
a	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
screen	NN	O	O
.	NN	O	O

Tap	NN	O	B-protein
associated	NN	O	O
with	NN	O	O
Tip	NN	O	B-protein
following	NN	O	O
transient	NN	O	O
expression	NN	O	O
in	NN	O	O
COS-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
stable	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
Jurkat-T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
Tip	NN	O	B-protein
and	NN	O	O
Tap	NN	O	B-protein
in	NN	O	O
Jurkat-T	NN	O	B-cell_line
cells	NN	O	I-cell_line
induced	NN	O	O
dramatic	NN	O	O
cell	NN	O	O
aggregation	NN	O	O
.	NN	O	O

Aggregation	NN	O	O
was	NN	O	O
likely	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
up-regulated	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
including	NN	O	O
integrin	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
L-selectin	NN	O	B-protein
,	NN	O	O
ICAM-3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
H-CAM	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
transcriptional	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	O
aggregated	NN	O	O
cells	NN	O	O
had	NN	O	O
approximately	NN	O	O
40-fold	NN	O	O
higher	NN	O	O
activity	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
parental	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Tap	NN	O	B-protein
is	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
cellular	NN	O	O
mediator	NN	O	O
of	NN	O	O
Tip	NN	O	B-protein
function	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
transformation	NN	O	O
by	NN	O	O
herpesvirus	NN	O	O
saimiri	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-cell_type
monocyte	NN	O	I-cell_type
binding	NN	O	O
to	NN	O	O
fibronectin	NN	O	B-protein
enhances	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
early	NN	O	O
signaling	NN	O	O
events	NN	O	O
.	NN	O	O

Leukocyte	NN	O	B-protein
integrins	NN	O	I-protein
are	NN	O	O
fundamentally	NN	O	O
important	NN	O	O
in	NN	O	O
modulating	NN	O	O
adhesion	NN	O	O
to	NN	O	O
extracellular	NN	O	O
matrix	NN	O	O
components	NN	O	O
and	NN	O	O
to	NN	O	O
other	NN	O	O
cells	NN	O	O
.	NN	O	O

This	NN	O	O
integrin-mediated	NN	O	O
adhesion	NN	O	O
controls	NN	O	O
leukocyte	NN	O	O
arrest	NN	O	O
and	NN	O	O
extravasation	NN	O	O
during	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
integrin	NN	O	B-protein
-ligand	NN	O	O
interactions	NN	O	O
trigger	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
that	NN	O	O
may	NN	O	O
influence	NN	O	O
leukocyte	NN	O	O
phenotype	NN	O	O
and	NN	O	O
function	NN	O	O
at	NN	O	O
sites	NN	O	O
of	NN	O	O
inflammation	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
current	NN	O	O
studies	NN	O	O
,	NN	O	O
we	NN	O	O
evaluated	NN	O	O
the	NN	O	O
combinatorial	NN	O	O
effects	NN	O	O
of	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
on	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

IFN-gamma	NN	O	B-protein
triggers	NN	O	O
a	NN	O	O
well-defined	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
,	NN	O	O
which	NN	O	O
although	NN	O	O
not	NN	O	O
directly	NN	O	O
stimulated	NN	O	O
by	NN	O	O
monocyte	NN	O	O
adherence	NN	O	O
to	NN	O	O
fibronectin	NN	O	B-protein
or	NN	O	O
arginine-glycine-aspartate	NN	O	B-protein
(	NN	O	I-protein
RGD	NN	O	I-protein
)	NN	O	I-protein
-coated	NN	O	I-protein
substrata	NN	O	I-protein
,	NN	O	O
was	NN	O	O
enhanced	NN	O	O
significantly	NN	O	O
in	NN	O	O
these	NN	O	O
matrix-adherent	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Compared	NN	O	O
with	NN	O	O
monocytes	NN	O	O
in	NN	O	O
suspension	NN	O	O
or	NN	O	O
adherent	NN	O	O
on	NN	O	O
plastic	NN	O	O
surfaces	NN	O	O
,	NN	O	O
monocytes	NN	O	O
adherent	NN	O	O
to	NN	O	O
fibronectin	NN	O	B-protein
or	NN	O	O
RGD	NN	O	O
exhibited	NN	O	O
a	NN	O	O
greater	NN	O	O
than	NN	O	O
threefold	NN	O	O
increase	NN	O	O
in	NN	O	O
steady	NN	O	O
state	NN	O	O
levels	NN	O	O
of	NN	O	O
IFN-gamma-induced	NN	O	B-RNA
mRNA	NN	O	I-RNA
for	NN	O	O
the	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
Fc	NN	O	I-protein
gammaRI	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

By	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
this	NN	O	O
increase	NN	O	O
in	NN	O	O
mRNA	NN	O	B-RNA
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
5-	NN	O	O
to	NN	O	O
10-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
STAT1-containing	NN	O	B-protein
DNA-binding	NN	O	I-protein
complex	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
to	NN	O	O
Fc	NN	O	B-DNA
gammaRI	NN	O	I-DNA
promoter	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
JAK1	NN	O	B-protein
and	NN	O	O
JAK2	NN	O	B-protein
was	NN	O	O
enhanced	NN	O	O
significantly	NN	O	O
in	NN	O	O
RGD-adherent	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
compared	NN	O	O
with	NN	O	O
control	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
novel	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
integrin	NN	O	B-protein
-mediated	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
can	NN	O	O
modulate	NN	O	O
the	NN	O	O
magnitude	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
,	NN	O	O
thereby	NN	O	O
amplifying	NN	O	O
cellular	NN	O	O
events	NN	O	O
leading	NN	O	O
to	NN	O	O
monocyte	NN	O	O
activation	NN	O	O
and	NN	O	O
inflammation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Surfactant	NN	O	B-protein
protein	NN	O	I-protein
A	NN	O	I-protein
activates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
THP-1	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
genes	NN	O	O
for	NN	O	O
which	NN	O	O
products	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
inflammation	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
regulator	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Because	NN	O	O
surfactant	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
SP	NN	O	I-protein
)	NN	O	I-protein
A	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
local	NN	O	O
host	NN	O	O
defense	NN	O	O
in	NN	O	O
the	NN	O	O
lung	NN	O	O
and	NN	O	O
alters	NN	O	O
immune	NN	O	O
cell	NN	O	O
function	NN	O	O
by	NN	O	O
modulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
surface	NN	O	B-protein
proteins	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
inflammation	NN	O	O
,	NN	O	O
we	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
SP-A	NN	O	B-protein
exerts	NN	O	O
its	NN	O	O
action	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
via	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

We	NN	O	O
used	NN	O	O
gel	NN	O	O
shift	NN	O	O
assays	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
SP-A	NN	O	B-protein
activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
THP-1	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
SP-A	NN	O	B-protein
doses	NN	O	O
as	NN	O	O
low	NN	O	O
as	NN	O	O
1	NN	O	O
microgram/ml	NN	O	O
occurred	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
of	NN	O	O
SP-A	NN	O	B-protein
treatment	NN	O	O
,	NN	O	O
peaked	NN	O	O
at	NN	O	O
60	NN	O	O
min	NN	O	O
,	NN	O	O
and	NN	O	O
then	NN	O	O
declined	NN	O	O
.	NN	O	O

This	NN	O	O
activation	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
known	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
or	NN	O	O
by	NN	O	O
simultaneous	NN	O	O
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
surfactant	NN	O	O
lipids	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibitors	NN	O	I-protein
blocked	NN	O	O
SP-A	NN	O	B-protein
-dependent	NN	O	O
increases	NN	O	O
in	NN	O	O
tumor	NN	O	B-RNA
necrosis	NN	O	I-RNA
factor-alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
a	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
SP-A	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
some	NN	O	O
lung	NN	O	O
conditions	NN	O	O
and	NN	O	O
point	NN	O	O
to	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
measures	NN	O	O
that	NN	O	O
could	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
prevent	NN	O	O
SP-A	NN	O	B-protein
induced	NN	O	O
inflammation	NN	O	O
in	NN	O	O
the	NN	O	O
lung	NN	O	O
.	NN	O	O

-DOCSTART-	O

GABP	NN	O	B-protein
factors	NN	O	I-protein
bind	NN	O	O
to	NN	O	O
a	NN	O	O
distal	NN	O	B-DNA
interleukin	NN	O	I-DNA
2	NN	O	I-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
contribute	NN	O	O
to	NN	O	O
c-Raf	NN	O	B-protein
-mediated	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

Triggering	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
receptor-CD3	NN	O	I-protein
complex	NN	O	I-protein
activates	NN	O	O
two	NN	O	O
major	NN	O	O
signal	NN	O	O
cascades	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
Ca2+-dependent	NN	O	O
signal	NN	O	O
cascades	NN	O	O
and	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
cascades	NN	O	O
.	NN	O	O

Both	NN	O	O
signal	NN	O	O
cascades	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Prominent	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
cascades	NN	O	O
are	NN	O	O
those	NN	O	O
that	NN	O	O
activate	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MAP	NN	O	I-protein
)	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
c-Raf	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
at	NN	O	O
the	NN	O	O
helm	NN	O	O
of	NN	O	O
the	NN	O	O
classic	NN	O	O
MAP-	NN	O	O
Erk	NN	O	B-protein
cascade	NN	O	O
,	NN	O	O
contributes	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
induction	NN	O	O
through	NN	O	O
a	NN	O	O
distal	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
spanning	NN	O	O
the	NN	O	O
nucleotides	NN	O	O
from	NN	O	O
positions	NN	O	O
-502	NN	O	O
to	NN	O	O
-413	NN	O	O
in	NN	O	O
front	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
start	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
this	NN	O	O
distal	NN	O	B-DNA
IL-2	NN	O	I-DNA
enhancer	NN	O	I-DNA
differs	NN	O	O
from	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
IL-2	NN	O	I-DNA
promoter-enhancer	NN	O	I-DNA
,	NN	O	O
since	NN	O	O
it	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
alone	NN	O	O
and	NN	O	O
independent	NN	O	O
from	NN	O	O
Ca2+	NN	O	O
signals	NN	O	O
.	NN	O	O

In	NN	O	O
DNA-protein	NN	O	O
binding	NN	O	O
studies	NN	O	O
,	NN	O	O
we	NN	O	O
detected	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
GABP	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
-beta	NN	O	I-protein
to	NN	O	O
a	NN	O	O
dyad	NN	O	B-DNA
symmetry	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
DSE	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
formed	NN	O	O
by	NN	O	O
palindromic	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
of	NN	O	O
Ets-like	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Introduction	NN	O	O
of	NN	O	O
point	NN	O	O
mutations	NN	O	O
suppressing	NN	O	O
GABP	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
DSE	NN	O	B-DNA
interfered	NN	O	O
with	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
entire	NN	O	O
IL-2	NN	O	B-DNA
promoter-enhancer	NN	O	I-DNA
,	NN	O	O
while	NN	O	O
overexpression	NN	O	O
of	NN	O	O
both	NN	O	O
GABP	NN	O	B-protein
factors	NN	O	I-protein
enhanced	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter-enhancer	NN	O	I-DNA
induction	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
BXB	NN	O	B-protein
,	NN	O	O
a	NN	O	O
constitutive	NN	O	O
active	NN	O	O
version	NN	O	O
of	NN	O	O
c-Raf	NN	O	B-protein
,	NN	O	O
and	NN	O	O
of	NN	O	O
further	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Ras	NN	O	B-protein
-	NN	O	O
Raf	NN	O	B-protein
-	NN	O	O
Erk	NN	O	B-protein
signal	NN	O	O
cascade	NN	O	O
exerted	NN	O	O
an	NN	O	O
increase	NN	O	O
of	NN	O	O
GABP	NN	O	B-protein
-mediated	NN	O	O
promoter-enhancer	NN	O	O
induction	NN	O	O
.	NN	O	O

In	NN	O	O
conjunction	NN	O	O
with	NN	O	O
previously	NN	O	O
published	NN	O	O
data	NN	O	O
on	NN	O	O
c-Raf	NN	O	B-protein
-induced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
GABP	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
E.Flory	NN	O	O
,	NN	O	O
A.	NN	O	O
Hoffmeyer	NN	O	O
,	NN	O	O
U.Smola	NN	O	O
,	NN	O	O
U.R.Rapp	NN	O	O
,	NN	O	O
and	NN	O	O
J.T.Bruder	NN	O	O
,	NN	O	O
J.Virol.70	NN	O	O
:	NN	O	O
2260-	NN	O	O
2268	NN	O	O
,	NN	O	O
1996	NN	O	O
)	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
indicate	NN	O	O
a	NN	O	O
contribution	NN	O	O
of	NN	O	O
GABP	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
Raf	NN	O	B-protein
-mediated	NN	O	O
enhancement	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
induction	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lipopolysaccharide	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor-alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Regulation	NN	O	O
by	NN	O	O
Egr-1	NN	O	B-protein
,	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Biosynthesis	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
is	NN	O	O
predominantly	NN	O	O
by	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
study	NN	O	O
examined	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
various	NN	O	O
cis-acting	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
TNF-alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
cells	NN	O	O
of	NN	O	O
monocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

Functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
monocytic	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
transfected	NN	O	O
with	NN	O	O
plasmids	NN	O	B-DNA
containing	NN	O	O
various	NN	O	O
lengths	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
localized	NN	O	O
enhancer	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
region	NN	O	O
(	NN	O	O
-182	NN	O	B-DNA
to	NN	O	I-DNA
-37	NN	O	I-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
(	NN	O	O
bp	NN	O	O
)	NN	O	O
)	NN	O	O
that	NN	O	O
were	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
.	NN	O	O

Two	NN	O	O
regions	NN	O	O
were	NN	O	O
identified	NN	O	O
:	NN	O	O
region	NN	O	B-DNA
I	NN	O	I-DNA
(	NN	O	O
-182	NN	O	B-DNA
to	NN	O	I-DNA
-162	NN	O	I-DNA
bp	NN	O	I-DNA
)	NN	O	O
contained	NN	O	O
an	NN	O	O
overlapping	NN	O	O
Sp1/Egr-1	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
region	NN	O	B-DNA
II	NN	O	I-DNA
(	NN	O	O
-119	NN	O	B-DNA
to	NN	O	I-DNA
-88	NN	O	I-DNA
)	NN	O	O
contained	NN	O	O
CRE	NN	O	B-DNA
and	NN	O	O
NF-kappaB	NN	O	B-DNA
(	NN	O	I-DNA
designated	NN	O	I-DNA
kappaB3	NN	O	I-DNA
)	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
unstimulated	NN	O	O
THP-1	NN	O	B-cell_line
,	NN	O	O
CRE-binding	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
c-Jun	NN	O	B-protein
complexes	NN	O	I-protein
were	NN	O	O
found	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

LPS	NN	O	O
stimulation	NN	O	O
increased	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
c-Jun-containing	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
induced	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
cognate	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
Egr-1	NN	O	B-DNA
and	NN	O	I-DNA
kappaB3	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
were	NN	O	O
identified	NN	O	O
as	NN	O	O
Egr-1	NN	O	B-protein
and	NN	O	O
p50/p65	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
CRE	NN	O	B-DNA
and	NN	O	I-DNA
kappaB3	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
region	NN	O	B-DNA
II	NN	O	I-DNA
together	NN	O	O
conferred	NN	O	O
strong	NN	O	O
LPS	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
whereas	NN	O	O
individually	NN	O	O
they	NN	O	O
failed	NN	O	O
to	NN	O	O
provide	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
increasing	NN	O	O
the	NN	O	O
spacing	NN	O	O
between	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
kappaB3	NN	O	I-DNA
sites	NN	O	I-DNA
completely	NN	O	O
abolished	NN	O	O
LPS	NN	O	O
induction	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
cooperative	NN	O	O
interaction	NN	O	O
between	NN	O	O
c-Jun	NN	O	B-protein
complexes	NN	O	I-protein
and	NN	O	O
p50/p65	NN	O	B-protein
.	NN	O	O

These	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
maximal	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
concerted	NN	O	O
participation	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
separate	NN	O	O
cis-acting	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Rescue	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
of	NN	O	O
apoptotic	NN	O	O
cell	NN	O	O
death	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
deprivation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
antigen-specific	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
control	NN	O	O
of	NN	O	O
cell	NN	O	O
survival	NN	O	O
and	NN	O	O
cell	NN	O	O
death	NN	O	O
is	NN	O	O
of	NN	O	O
central	NN	O	O
importance	NN	O	O
in	NN	O	O
tissues	NN	O	O
with	NN	O	O
high	NN	O	O
cell	NN	O	O
turnover	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
lymphoid	NN	O	O
system	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
on	NN	O	O
IL-2	NN	O	B-protein
deprivation-induced	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
antigen-specific	NN	O	I-cell_line
T	NN	O	I-cell_line
helper	NN	O	I-cell_line
clones	NN	O	I-cell_line
with	NN	O	O
different	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
profiles	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
interferon-alpha	NN	O	B-protein
(	NN	O	O
IFN-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
IFN-beta	NN	O	B-protein
inhibited	NN	O	O
IL-2	NN	O	B-protein
deprivation	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
Th0	NN	O	B-cell_line
,	NN	O	I-cell_line
Th1	NN	O	I-cell_line
,	NN	O	I-cell_line
and	NN	O	I-cell_line
Th2	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
also	NN	O	O
found	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
protects	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
from	NN	O	O
IL-2	NN	O	B-protein
deprivation	NN	O	O
apoptosis	NN	O	O
accompanying	NN	O	O
active	NN	O	O
proliferation	NN	O	O
and	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
P53	NN	O	B-protein
,	NN	O	O
Rb	NN	O	B-protein
and	NN	O	O
Bcl-xL	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
IFN-alpha/beta	NN	O	B-protein
rescued	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
from	NN	O	O
apoptosis	NN	O	O
without	NN	O	O
active	NN	O	O
proliferation	NN	O	O
,	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
apoptosis-associated	NN	O	B-protein
proteins	NN	O	I-protein
tested	NN	O	O
so	NN	O	O
far	NN	O	O
was	NN	O	O
unaffected	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
contained	NN	O	O
those	NN	O	O
located	NN	O	O
in	NN	O	O
S	NN	O	O
+	NN	O	O
G2/M	NN	O	O
phases	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
vast	NN	O	O
majority	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
IFN-alpha/beta	NN	O	B-protein
were	NN	O	O
located	NN	O	O
in	NN	O	O
G0/G1	NN	O	O
phase	NN	O	O
.	NN	O	O

IFN-alpha/beta	NN	O	B-protein
specifically	NN	O	O
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
translocation	NN	O	O
into	NN	O	O
nucleus	NN	O	O
of	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
2	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
over-expression	NN	O	O
of	NN	O	O
STAT2	NN	O	B-protein
by	NN	O	O
transfection	NN	O	O
of	NN	O	O
the	NN	O	O
cDNA	NN	O	O
prevented	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
present	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
IFN-alpha	NN	O	B-protein
and	NN	O	I-protein
-beta	NN	O	I-protein
mediate	NN	O	O
anti-apoptotic	NN	O	O
effect	NN	O	O
through	NN	O	O
other	NN	O	O
pathways	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
growth	NN	O	O
factor	NN	O	O
deprivation	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin-10	NN	O	B-protein
inhibits	NN	O	O
interferon-gamma-induced	NN	O	O
intercellular	NN	O	B-DNA
adhesion	NN	O	I-DNA
molecule-1	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Interleukin-10	NN	O	B-protein
(	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
monocyte	NN	O	B-protein
regulatory	NN	O	I-protein
cytokine	NN	O	I-protein
that	NN	O	O
inhibits	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
mediators	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
IL-10	NN	O	B-protein
downregulates	NN	O	O
expression	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
activated	NN	O	O
with	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
inhibition	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
apparent	NN	O	O
as	NN	O	O
early	NN	O	O
as	NN	O	O
3	NN	O	O
hours	NN	O	O
and	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
an	NN	O	O
anti-IL-10	NN	O	B-protein
antibody	NN	O	I-protein
but	NN	O	O
not	NN	O	O
by	NN	O	O
an	NN	O	O
isotype-matched	NN	O	B-protein
control	NN	O	I-protein
antibody	NN	O	I-protein
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
reduced	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
IFN-gamma-stimulated	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
.	NN	O	O

IL-10	NN	O	B-protein
inhibition	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-RNA
steady-state	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
detected	NN	O	O
at	NN	O	O
3	NN	O	O
hours	NN	O	O
and	NN	O	O
remained	NN	O	O
at	NN	O	O
24	NN	O	O
hours	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
run-on	NN	O	O
transcription	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
inhibited	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
did	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
half-life	NN	O	O
of	NN	O	O
IFN-gamma-induced	NN	O	B-RNA
ICAM-1	NN	O	I-RNA
message	NN	O	I-RNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
inhibits	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
primarily	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
IFN-gamma-responsive	NN	O	B-DNA
genes	NN	O	I-DNA
requires	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
factor	NN	O	O
STAT-1alpha	NN	O	B-protein
(	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription-1alpha	NN	O	I-protein
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT-1alpha	NN	O	B-protein
or	NN	O	O
alter	NN	O	O
STAT-1alpha	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
IRE	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Instead	NN	O	O
,	NN	O	O
IL-10	NN	O	B-protein
prevented	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
binding	NN	O	O
activity	NN	O	O
at	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
site	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
TNF-alpha	NN	O	I-DNA
)	NN	O	I-DNA
-responsive	NN	O	I-DNA
NF-kappaB/C-EBP	NN	O	I-DNA
composite	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
inhibits	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
ICAM-1	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
a	NN	O	O
regulatory	NN	O	O
mechanism	NN	O	O
that	NN	O	O
may	NN	O	O
involve	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Specific	NN	O	O
complex	NN	O	O
formation	NN	O	O
between	NN	O	O
the	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
bare	NN	O	I-protein
lymphocyte	NN	O	I-protein
syndrome-associated	NN	O	I-protein
transactivators	NN	O	I-protein
CIITA	NN	O	B-protein
and	NN	O	O
RFX5	NN	O	B-protein
.	NN	O	O

Two	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
defective	NN	O	O
in	NN	O	O
the	NN	O	O
five	NN	O	O
complementation	NN	O	O
groups	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
class	NN	O	O
II-negative	NN	O	O
bare	NN	O	O
lymphocyte	NN	O	O
syndrome	NN	O	O
or	NN	O	O
corresponding	NN	O	O
laboratory	NN	O	O
mutants	NN	O	O
have	NN	O	O
been	NN	O	O
cloned	NN	O	O
.	NN	O	O

One	NN	O	O
gene	NN	O	O
encodes	NN	O	O
a	NN	O	O
protein	NN	O	O
,	NN	O	O
RFX5	NN	O	B-protein
,	NN	O	O
that	NN	O	O
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
RFX	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
other	NN	O	O
,	NN	O	O
CIITA	NN	O	B-protein
,	NN	O	O
encodes	NN	O	O
a	NN	O	O
large	NN	O	O
protein	NN	O	O
with	NN	O	O
a	NN	O	O
defined	NN	O	O
acidic	NN	O	B-protein
transcriptional	NN	O	I-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
;	NN	O	O
this	NN	O	O
protein	NN	O	O
does	NN	O	O
not	NN	O	O
interact	NN	O	O
with	NN	O	O
DNA	NN	O	O
.	NN	O	O

Expression	NN	O	B-DNA
plasmids	NN	O	I-DNA
encoding	NN	O	O
regions	NN	O	O
of	NN	O	O
RFX5	NN	O	B-protein
fused	NN	O	O
to	NN	O	O
the	NN	O	O
GAL4	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
activated	NN	O	O
transcription	NN	O	O
from	NN	O	O
a	NN	O	O
reporter	NN	O	O
construct	NN	O	O
containing	NN	O	O
GAL4	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
cotransfection	NN	O	O
assay	NN	O	O
in	NN	O	O
the	NN	O	O
Raji	NN	O	B-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
these	NN	O	O
plasmids	NN	O	B-DNA
produced	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
only	NN	O	O
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
stimulation	NN	O	O
,	NN	O	O
a	NN	O	O
condition	NN	O	O
in	NN	O	O
which	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
CIITA	NN	O	B-protein
and	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
major	NN	O	I-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
surface	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
induced	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
these	NN	O	O
plasmids	NN	O	B-DNA
were	NN	O	O
not	NN	O	O
active	NN	O	O
in	NN	O	O
RJ2.2.5	NN	O	B-cell_line
,	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
mutagenized	NN	O	O
derivative	NN	O	O
of	NN	O	O
Raji	NN	O	B-cell_line
in	NN	O	O
which	NN	O	O
both	NN	O	O
copies	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
are	NN	O	O
defective	NN	O	O
.	NN	O	O

Transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
the	NN	O	O
RFX5	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
could	NN	O	O
be	NN	O	O
restored	NN	O	O
in	NN	O	O
RJ2.2.5	NN	O	B-cell_line
by	NN	O	O
cotransfection	NN	O	O
with	NN	O	O
a	NN	O	O
CIITA	NN	O	B-DNA
expression	NN	O	I-DNA
plasmid	NN	O	I-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
a	NN	O	O
direct	NN	O	O
interaction	NN	O	O
between	NN	O	O
RFX5	NN	O	B-protein
and	NN	O	O
CIITA	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
with	NN	O	O
the	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
and	NN	O	O
far-Western	NN	O	O
blot	NN	O	O
assays	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
RFX5	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
transcription	NN	O	O
only	NN	O	O
in	NN	O	O
cooperation	NN	O	O
with	NN	O	O
CIITA	NN	O	B-protein
.	NN	O	O

RFX5	NN	O	B-protein
and	NN	O	O
CIITA	NN	O	B-protein
associate	NN	O	O
to	NN	O	O
form	NN	O	O
a	NN	O	O
complex	NN	O	O
capable	NN	O	O
of	NN	O	O
activating	NN	O	O
transcription	NN	O	O
from	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
major	NN	O	I-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
complex	NN	O	O
,	NN	O	O
promoter	NN	O	O
specificity	NN	O	O
is	NN	O	O
determined	NN	O	O
by	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
domain	NN	O	O
of	NN	O	O
RFX5	NN	O	B-protein
and	NN	O	O
the	NN	O	O
general	NN	O	O
transcription	NN	O	O
apparatus	NN	O	O
is	NN	O	O
recruited	NN	O	O
by	NN	O	O
the	NN	O	O
acidic	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
CIITA	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
in	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
(	NN	O	O
PI3-K	NN	O	B-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
in	NN	O	O
many	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
the	NN	O	O
PI3-K	NN	O	B-protein
inhibitor	NN	O	O
,	NN	O	O
wortmannin	NN	O	O
,	NN	O	O
interferes	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
Erk2	NN	O	B-protein
,	NN	O	O
after	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TcR	NN	O	B-protein
)	NN	O	O
stimulation	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
explore	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
PI3-K	NN	O	B-protein
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
we	NN	O	O
created	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
potentially	NN	O	O
dominant	NN	O	B-DNA
negative	NN	O	I-DNA
PI3-K	NN	O	I-DNA
constructs	NN	O	I-DNA
comprising	NN	O	O
individual	NN	O	O
or	NN	O	O
tandem	NN	O	B-DNA
domains	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
regulatory	NN	O	B-protein
p85	NN	O	I-protein
subunit	NN	O	I-protein
and	NN	O	O
tested	NN	O	O
their	NN	O	O
effect	NN	O	O
on	NN	O	O
downstream	NN	O	O
signaling	NN	O	O
events	NN	O	O
like	NN	O	O
Erk2	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
transcription	NN	O	O
from	NN	O	O
an	NN	O	O
NFAT	NN	O	B-DNA
(	NN	O	I-DNA
nuclear	NN	O	I-DNA
factor	NN	O	I-DNA
of	NN	O	I-DNA
activated	NN	O	I-DNA
T	NN	O	I-DNA
cells	NN	O	I-DNA
)	NN	O	I-DNA
element	NN	O	I-DNA
taken	NN	O	O
from	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Following	NN	O	O
TcR	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
Erk2	NN	O	B-protein
was	NN	O	O
only	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
previously	NN	O	O
described	NN	O	O
truncated	NN	O	B-protein
form	NN	O	I-protein
of	NN	O	I-protein
p85	NN	O	I-protein
that	NN	O	O
can	NN	O	O
not	NN	O	O
bind	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
subunit	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
other	NN	O	O
constructs	NN	O	O
of	NN	O	O
p85	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
several	NN	O	O
mutant	NN	O	O
p85	NN	O	B-protein
alleles	NN	O	O
had	NN	O	O
dramatic	NN	O	O
effects	NN	O	O
on	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Most	NN	O	O
interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
SH2	NN	O	I-protein
domain	NN	O	I-protein
had	NN	O	O
an	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
mutant	NN	O	B-protein
p85	NN	O	I-protein
containing	NN	O	O
only	NN	O	O
the	NN	O	O
two	NN	O	B-protein
SH2	NN	O	I-protein
domains	NN	O	I-protein
enhanced	NN	O	O
basal	NN	O	O
NFAT	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
a	NN	O	O
Ras	NN	O	B-protein
-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Ionomycin	NN	O	O
induced	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
in	NN	O	O
cells	NN	O	O
expressing	NN	O	O
p85	NN	O	B-protein
mutants	NN	O	I-protein
that	NN	O	O
contained	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
SH2	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
phosphotyrosine-containing	NN	O	B-protein
proteins	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
truncated	NN	O	O
p85	NN	O	B-protein
constructs	NN	O	I-protein
revealed	NN	O	O
cooperative	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	B-protein
SH2	NN	O	I-protein
domains	NN	O	I-protein
but	NN	O	O
no	NN	O	O
apparent	NN	O	O
differences	NN	O	O
between	NN	O	O
the	NN	O	O
N-	NN	O	B-protein
and	NN	O	I-protein
C-	NN	O	I-protein
terminal	NN	O	I-protein
SH2	NN	O	I-protein
domains	NN	O	I-protein
.	NN	O	O

Wortmannin	NN	O	O
did	NN	O	O
not	NN	O	O
interfere	NN	O	O
with	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
although	NN	O	O
it	NN	O	O
inhibited	NN	O	O
PI3-K	NN	O	B-protein
and	NN	O	O
Erk2	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
experiment	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
PI3-K	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
through	NN	O	O
a	NN	O	O
complex	NN	O	O
adaptor	NN	O	O
function	NN	O	O
of	NN	O	O
its	NN	O	O
regulatory	NN	O	B-protein
subunit	NN	O	I-protein
and	NN	O	O
that	NN	O	O
its	NN	O	O
lipid	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
is	NN	O	O
dispensable	NN	O	O
for	NN	O	O
this	NN	O	O
effect	NN	O	O
.	NN	O	O

-DOCSTART-	O

Acute	NN	O	O
leukemia	NN	O	O
with	NN	O	O
promyelocytic	NN	O	O
features	NN	O	O
in	NN	O	O
PML/RARalpha	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
reciprocal	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
involving	NN	O	O
the	NN	O	O
retinoic	NN	O	B-DNA
acid	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
RARalpha	NN	O	I-DNA
)	NN	O	I-DNA
locus	NN	O	I-DNA
on	NN	O	O
chromosome	NN	O	B-DNA
17	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
cases	NN	O	O
,	NN	O	O
RARalpha	NN	O	B-protein
translocates	NN	O	O
and	NN	O	O
fuses	NN	O	O
with	NN	O	O
the	NN	O	O
promyelocytic	NN	O	B-DNA
leukemia	NN	O	I-DNA
(	NN	O	I-DNA
PML	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
located	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
15	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
resulting	NN	O	O
fusion	NN	O	B-DNA
genes	NN	O	I-DNA
encode	NN	O	O
the	NN	O	O
two	NN	O	O
structurally	NN	O	O
unique	NN	O	O
PML/RARalpha	NN	O	B-protein
and	NN	O	O
RARalpha/PML	NN	O	B-protein
fusion	NN	O	I-protein
proteins	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
aberrant	NN	O	B-protein
PML	NN	O	I-protein
gene	NN	O	I-protein
products	NN	O	I-protein
,	NN	O	O
the	NN	O	O
respective	NN	O	O
pathogenetic	NN	O	O
roles	NN	O	O
of	NN	O	O
which	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
elucidated	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
generated	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
PML/RARalpha	NN	O	B-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
specifically	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
myeloid-promyelocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

During	NN	O	O
their	NN	O	O
first	NN	O	O
year	NN	O	O
of	NN	O	O
life	NN	O	O
,	NN	O	O
all	NN	O	O
the	NN	O	O
PML/	NN	O	O
RARalpha	NN	O	B-protein
transgenic	NN	O	O
mice	NN	O	O
have	NN	O	O
an	NN	O	O
abnormal	NN	O	O
hematopoiesis	NN	O	O
that	NN	O	O
can	NN	O	O
best	NN	O	O
be	NN	O	O
described	NN	O	O
as	NN	O	O
a	NN	O	O
myeloproliferative	NN	O	O
disorder	NN	O	O
.	NN	O	O

Between	NN	O	O
12	NN	O	O
and	NN	O	O
14	NN	O	O
months	NN	O	O
of	NN	O	O
age	NN	O	O
,	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
them	NN	O	O
develop	NN	O	O
a	NN	O	O
form	NN	O	O
of	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
with	NN	O	O
a	NN	O	O
differentiation	NN	O	O
block	NN	O	O
at	NN	O	O
the	NN	O	O
promyelocytic	NN	O	O
stage	NN	O	O
that	NN	O	O
closely	NN	O	O
mimics	NN	O	O
human	NN	O	O
APL	NN	O	O
even	NN	O	O
in	NN	O	O
its	NN	O	O
response	NN	O	O
to	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
are	NN	O	O
conclusive	NN	O	O
in	NN	O	O
vivo	NN	O	O
evidence	NN	O	O
that	NN	O	O
PML/RARalpha	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
APL	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
protein/DNA	NN	O	O
interactions	NN	O	O
of	NN	O	O
the	NN	O	O
interferon-gamma	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
[	NN	O	O
corrected	NN	O	O
]	NN	O	O
CD45RA	NN	O	B-cell_type
and	NN	O	I-cell_type
CD45R0	NN	O	I-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
subsets	NN	O	I-cell_type
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
Eur	NN	O	O
J	NN	O	O
Immunol	NN	O	O
1997	NN	O	O
Jul	NN	O	O
;	NN	O	O
27	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
:	NN	O	O
1830	NN	O	O
]	NN	O	O

Interferon-gamma	NN	O	O
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
key	NN	O	O
cytokine	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
major	NN	O	O
regulatory	NN	O	O
functions	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
and	NN	O	O
compare	NN	O	O
protein/DNA	NN	O	O
interactions	NN	O	O
at	NN	O	O
the	NN	O	O
native	NN	O	O
IFN-gamma	NN	O	B-protein
locus	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IFN-gamma	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
ligation-mediated	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
LM-PCR	NN	O	O
)	NN	O	O
techniques	NN	O	O
.	NN	O	O

Accordingly	NN	O	O
,	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
primary	NN	O	B-cell_type
CD45RA	NN	O	I-cell_type
and	NN	O	I-cell_type
CD45R0	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
subsets	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
using	NN	O	O
immunomagnetic	NN	O	O
beads	NN	O	O
were	NN	O	O
cultured	NN	O	O
and	NN	O	O
analyzed	NN	O	O
by	NN	O	O
LM-PCR	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
protein/DNA	NN	O	O
interactions	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
vivo	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
all	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
tested	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
an	NN	O	O
inducible	NN	O	O
footprint	NN	O	O
between	NN	O	O
-183	NN	O	B-DNA
and	NN	O	I-DNA
-196	NN	O	I-DNA
was	NN	O	O
consistently	NN	O	O
observed	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
CD45RA	NN	O	B-cell_type
and	NN	O	I-cell_type
CD45R0	NN	O	I-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
subsets	NN	O	I-cell_type
upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate+	NN	O	O
phytohemagglutinin	NN	O	B-protein
(	NN	O	O
PMA+	NN	O	O
PHA	NN	O	B-protein
)	NN	O	O
that	NN	O	O
was	NN	O	O
highly	NN	O	O
sensitive	NN	O	O
to	NN	O	O
treatment	NN	O	O
with	NN	O	O
corticosteroids	NN	O	O
.	NN	O	O

This	NN	O	O
novel	NN	O	B-DNA
target	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
denoted	NN	O	O
the	NN	O	O
C-site	NN	O	B-DNA
,	NN	O	O
was	NN	O	O
shown	NN	O	O
by	NN	O	O
several	NN	O	O
criteria	NN	O	O
,	NN	O	O
including	NN	O	O
cell	NN	O	O
distribution	NN	O	O
studies	NN	O	O
,	NN	O	O
stimulation	NN	O	O
experiments	NN	O	O
,	NN	O	O
supershift	NN	O	O
assays	NN	O	O
,	NN	O	O
and	NN	O	O
cross-competition	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
to	NN	O	O
bind	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
C-site	NN	O	B-DNA
that	NN	O	O
prevented	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
was	NN	O	O
sufficient	NN	O	O
strikingly	NN	O	O
to	NN	O	O
reduce	NN	O	O
inducible	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
CD45R0	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
the	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
IFN-gamma	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
vivo	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
protein/DNA	NN	O	O
interactions	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
a	NN	O	O
model	NN	O	O
where	NN	O	O
basal	NN	O	O
transcription	NN	O	O
is	NN	O	O
maintained	NN	O	O
by	NN	O	O
binding	NN	O	O
of	NN	O	O
various	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
whereas	NN	O	O
PMA+	NN	O	O
PHA	NN	O	B-protein
-inducible	NN	O	O
IFN-gamma	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
CD45R0	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
binding	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
to	NN	O	O
the	NN	O	O
C-site	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
domain	NN	O	O
requirements	NN	O	O
for	NN	O	O
disruption	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
latency	NN	O	O
by	NN	O	O
ZEBRA	NN	O	B-protein
.	NN	O	O

Latent	NN	O	O
infection	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
can	NN	O	O
be	NN	O	O
disrupted	NN	O	O
by	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
ZEBRA	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

ZEBRA	NN	O	B-protein
,	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
,	NN	O	O
initiates	NN	O	O
the	NN	O	O
EBV	NN	O	O
lytic	NN	O	O
cascade	NN	O	O
by	NN	O	O
activating	NN	O	O
viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

ZEBRA	NN	O	B-protein
is	NN	O	O
also	NN	O	O
indispensable	NN	O	O
for	NN	O	O
viral	NN	O	O
replication	NN	O	O
and	NN	O	O
binds	NN	O	O
directly	NN	O	O
to	NN	O	O
the	NN	O	O
EBV	NN	O	O
lytic	NN	O	O
origin	NN	O	O
of	NN	O	O
replication	NN	O	O
.	NN	O	O

The	NN	O	O
studies	NN	O	O
described	NN	O	O
herein	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
domain	NN	O	O
.	NN	O	O

ZEBRA	NN	O	B-protein
activation	NN	O	O
can	NN	O	O
be	NN	O	O
replaced	NN	O	O
by	NN	O	O
a	NN	O	O
heterologous	NN	O	B-protein
acidic	NN	O	I-protein
,	NN	O	I-protein
proline-rich	NN	O	I-protein
,	NN	O	I-protein
or	NN	O	I-protein
glutamine-rich	NN	O	I-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

ZEBRA	NN	O	B-protein
activation	NN	O	O
domain	NN	O	O
swap	NN	O	O
constructs	NN	O	O
retain	NN	O	O
ZEBRA	NN	O	B-protein
's	NN	O	O
native	NN	O	O
abilities	NN	O	O
to	NN	O	O
activate	NN	O	O
specific	NN	O	O
EBV	NN	O	B-DNA
promoters	NN	O	I-DNA
,	NN	O	O
to	NN	O	O
disrupt	NN	O	O
EBV	NN	O	O
latency	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
stimulate	NN	O	O
replication	NN	O	O
at	NN	O	O
the	NN	O	O
EBV	NN	O	B-DNA
lytic	NN	O	I-DNA
origin	NN	O	I-DNA
.	NN	O	O

Additional	NN	O	O
work	NN	O	O
,	NN	O	O
employing	NN	O	O
sequential	NN	O	O
and	NN	O	O
internal	NN	O	O
deletions	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-protein
's	NN	O	O
N-terminal	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
indicates	NN	O	O
that	NN	O	O
its	NN	O	O
separate	NN	O	O
activities	NN	O	O
are	NN	O	O
not	NN	O	O
attributable	NN	O	O
to	NN	O	O
specific	NN	O	O
subdomains	NN	O	B-protein
but	NN	O	O
are	NN	O	O
spread	NN	O	O
throughout	NN	O	O
its	NN	O	O
N	NN	O	B-protein
terminus	NN	O	I-protein
and	NN	O	O
therefore	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
inactivated	NN	O	O
by	NN	O	O
deleting	NN	O	O
localized	NN	O	O
regions	NN	O	O
.	NN	O	O

-DOCSTART-	O

CholecystokininB	NN	O	B-protein
receptor	NN	O	I-protein
from	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
lymphoblastic	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
involved	NN	O	O
in	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
-responsive	NN	O	O
gene	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
cholecystokinin	NN	O	B-protein
(	NN	O	I-protein
CCK	NN	O	I-protein
(	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
lymphoblastic	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
trophic	NN	O	O
effect	NN	O	O
resulting	NN	O	O
from	NN	O	O
activation	NN	O	O
of	NN	O	O
such	NN	O	O
a	NN	O	O
receptor	NN	O	O
by	NN	O	O
using	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
strategy	NN	O	O
.	NN	O	O

For	NN	O	O
this	NN	O	O
purpose	NN	O	O
,	NN	O	O
we	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
plasmid	NN	O	I-DNA
p	NN	O	I-DNA
[	NN	O	I-DNA
(	NN	O	I-DNA
TRE	NN	O	I-DNA
)	NN	O	I-DNA
3-tk-Luc	NN	O	I-DNA
]	NN	O	I-DNA
and	NN	O	O
found	NN	O	O
that	NN	O	O
CCK-8	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
dose-dependently	NN	O	O
induce	NN	O	O
luciferase	NN	O	B-protein
expression	NN	O	O
related	NN	O	O
to	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
with	NN	O	O
a	NN	O	O
maximal	NN	O	O
response	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
obtained	NN	O	O
with	NN	O	O
compounds	NN	O	O
known	NN	O	O
to	NN	O	O
activate	NN	O	O
AP-1	NN	O	B-protein
complex	NN	O	I-protein
(	NN	O	O
quantitatively	NN	O	O
,	NN	O	O
the	NN	O	O
same	NN	O	O
level	NN	O	O
of	NN	O	O
induction	NN	O	O
was	NN	O	O
obtained	NN	O	O
with	NN	O	O
1	NN	O	O
nM	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
,	NN	O	O
100	NN	O	O
microM	NN	O	O
diacylglycerol	NN	O	O
,	NN	O	O
or	NN	O	O
4	NN	O	O
nM	NN	O	O
epidermal	NN	O	O
growth	NN	O	O
factor	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
CCK	NN	O	B-protein
(	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
in	NN	O	O
such	NN	O	O
a	NN	O	O
stimulation	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
the	NN	O	O
inhibiting	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
selective	NN	O	O
CCK	NN	O	B-protein
(	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
antagonist	NN	O	O
PD-135	NN	O	O
,	NN	O	O
158	NN	O	O
.	NN	O	O

This	NN	O	O
effect	NN	O	O
was	NN	O	O
confirmed	NN	O	O
in	NN	O	O
COS-7	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
cDNA	NN	O	O
of	NN	O	O
CCK	NN	O	B-protein
(	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
cloned	NN	O	O
from	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
-dependent	NN	O	O
luciferase	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
tested	NN	O	O
two	NN	O	O
specific	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
serine/threonine	NN	O	B-protein
phosphatases-1	NN	O	I-protein
and	NN	O	I-protein
-2A	NN	O	I-protein
:	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
and	NN	O	O
calyculin	NN	O	O
A	NN	O	O
.	NN	O	O

These	NN	O	O
compounds	NN	O	O
strongly	NN	O	O
increased	NN	O	O
the	NN	O	O
phorbol-12-myristate-13-acetate	NN	O	O
response	NN	O	O
,	NN	O	O
whereas	NN	O	O
we	NN	O	O
have	NN	O	O
not	NN	O	O
observed	NN	O	O
a	NN	O	O
contribution	NN	O	O
of	NN	O	O
phosphatase	NN	O	O
inhibitors	NN	O	O
on	NN	O	O
a	NN	O	O
CCK-8-induced	NN	O	O
luciferase	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

To	NN	O	O
confirm	NN	O	O
that	NN	O	O
CCK	NN	O	B-protein
(	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
receptors	NN	O	I-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
AP-1	NN	O	B-protein
response	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
CCK-8	NN	O	O
effect	NN	O	O
on	NN	O	O
interleukin-2	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
a	NN	O	O
natural	NN	O	O
endogenous	NN	O	B-DNA
gene	NN	O	I-DNA
regulated	NN	O	O
by	NN	O	O
several	NN	O	O
factors	NN	O	O
,	NN	O	O
including	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
activated	NN	O	O
by	NN	O	O
phorbol-12-myristate-13-acetate	NN	O	O
and	NN	O	O
phytohemagglutinin	NN	O	B-protein
,	NN	O	O
CCK-8	NN	O	O
induced	NN	O	O
IL-2	NN	O	O
expression	NN	O	O
.	NN	O	O

This	NN	O	O
induction	NN	O	O
was	NN	O	O
abolished	NN	O	O
by	NN	O	O
PD-135	NN	O	O
,	NN	O	O
158	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
CCK-8	NN	O	O
exerts	NN	O	O
a	NN	O	O
trophic	NN	O	O
effect	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
through	NN	O	O
stimulation	NN	O	O
of	NN	O	O
CCK	NN	O	B-protein
(	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
receptors	NN	O	I-protein
by	NN	O	O
modulation	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
AP-1-regulated	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
conserved	NN	O	O
tissue-specific	NN	O	O
structure	NN	O	O
at	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
T-cell	NN	O	I-DNA
receptor	NN	O	I-DNA
beta-chain	NN	O	I-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
T-cell	NN	O	B-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
TCR	NN	O	I-DNA
)	NN	O	I-DNA
beta-chain	NN	O	I-DNA
promoters	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
characterized	NN	O	O
as	NN	O	O
nonstructured	NN	O	B-DNA
basal	NN	O	I-DNA
promoters	NN	O	I-DNA
that	NN	O	O
carry	NN	O	O
a	NN	O	O
single	NN	O	O
conserved	NN	O	O
ubiquitous	NN	O	B-DNA
cyclic	NN	O	I-DNA
AMP-responsive	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
investigation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
TCR	NN	O	I-DNA
beta	NN	O	I-DNA
gene	NN	O	I-DNA
uncovers	NN	O	O
a	NN	O	O
surprisingly	NN	O	O
complex	NN	O	O
and	NN	O	O
tissue-specific	NN	O	O
structure	NN	O	O
at	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
Vbeta	NN	O	I-DNA
8.1	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
core	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
(	NN	O	O
positions	NN	O	B-DNA
-42	NN	O	I-DNA
to	NN	O	I-DNA
+11	NN	O	I-DNA
)	NN	O	O
is	NN	O	O
recognized	NN	O	O
by	NN	O	O
the	NN	O	O
lymphoid	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
Ets-1	NN	O	B-protein
,	NN	O	O
LEF1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
AML1	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
CREB/ATF-1	NN	O	B-protein
,	NN	O	O
as	NN	O	O
is	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
gel	NN	O	O
shift	NN	O	O
and	NN	O	O
footprinting	NN	O	O
experiments	NN	O	O
.	NN	O	O

With	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
LEF1	NN	O	B-protein
,	NN	O	O
these	NN	O	O
factors	NN	O	O
activate	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Binding	NN	O	B-DNA
sites	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
core	NN	O	O
region	NN	O	O
show	NN	O	O
little	NN	O	O
conservation	NN	O	O
with	NN	O	O
consensus	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

Nonetheless	NN	O	O
,	NN	O	O
CREB	NN	O	B-protein
,	NN	O	O
Ets-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
AML1	NN	O	B-protein
bind	NN	O	O
and	NN	O	O
activate	NN	O	O
cooperatively	NN	O	O
and	NN	O	O
very	NN	O	O
efficiently	NN	O	O
through	NN	O	O
the	NN	O	O
nonconsensus	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Moderate	NN	O	O
ubiquitous	NN	O	O
activation	NN	O	O
is	NN	O	O
further	NN	O	O
induced	NN	O	O
by	NN	O	O
CREB/ATF	NN	O	B-protein
and	NN	O	I-protein
Sp1	NN	O	I-protein
factors	NN	O	I-protein
through	NN	O	O
proximal	NN	O	B-DNA
upstream	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
tissue-specific	NN	O	B-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
structure	NN	O	I-DNA
is	NN	O	O
apparently	NN	O	O
conserved	NN	O	O
in	NN	O	O
other	NN	O	O
T-cell-specifically	NN	O	B-DNA
expressed	NN	O	I-DNA
genes	NN	O	I-DNA
such	NN	O	O
as	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
both	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
have	NN	O	O
a	NN	O	O
complex	NN	O	O
tissue-specific	NN	O	O
structure	NN	O	O
whose	NN	O	O
functional	NN	O	O
interplay	NN	O	O
potentiates	NN	O	O
T-cell-specific	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
GM-CSF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
subunit	NN	O	I-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
both	NN	O	O
GM-CSF	NN	O	B-protein
-mediated	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
regulates	NN	O	O
differentiation	NN	O	O
,	NN	O	O
survival	NN	O	O
,	NN	O	O
and	NN	O	O
proliferation	NN	O	O
of	NN	O	O
colony-forming	NN	O	B-cell_line
unit-granulocyte-macrophage	NN	O	I-cell_line
progenitor	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
biologic	NN	O	O
actions	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
binding	NN	O	O
to	NN	O	O
a	NN	O	O
specific	NN	O	O
receptor	NN	O	O
consisting	NN	O	O
of	NN	O	O
two	NN	O	O
chains	NN	O	O
designated	NN	O	O
as	NN	O	O
alpha	NN	O	B-protein
and	NN	O	I-protein
beta	NN	O	I-protein
subunits	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
FDC-P1-derived	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
WT-19	NN	O	B-cell_line
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	B-protein
GM-CSF	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
beta	NN	O	I-protein
subunits	NN	O	I-protein
(	NN	O	O
GM-CSFRalpha	NN	O	B-protein
and	NN	O	I-protein
beta	NN	O	I-protein
)	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
human	NN	O	B-protein
GM-CSF	NN	O	I-protein
(	NN	O	O
hGM-CSF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

By	NN	O	O
expressing	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
GM-CSFRalpha	NN	O	B-protein
mutants	NN	O	I-protein
in	NN	O	O
WT19	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
have	NN	O	O
determined	NN	O	O
the	NN	O	O
amino	NN	O	B-protein
acid	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
the	NN	O	O
GM-CSFRalpha	NN	O	B-protein
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
proliferation	NN	O	O
,	NN	O	O
and	NN	O	O
survival	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
membrane	NN	O	B-protein
proximal	NN	O	I-protein
proline-rich	NN	O	I-protein
domain	NN	O	I-protein
and	NN	O	O
adjacent	NN	O	B-protein
16	NN	O	I-protein
residues	NN	O	I-protein
are	NN	O	O
essential	NN	O	O
for	NN	O	O
both	NN	O	O
hGM-CSF	NN	O	B-protein
-dependent	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
GM-CSFRalpha	NN	O	B-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
was	NN	O	O
not	NN	O	O
necessary	NN	O	O
for	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
mediated	NN	O	O
by	NN	O	O
hGM-CSF	NN	O	B-protein
,	NN	O	O
but	NN	O	O
the	NN	O	O
removal	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
severely	NN	O	O
impaired	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
hGM-CSF	NN	O	B-protein
to	NN	O	O
support	NN	O	O
cell	NN	O	O
survival	NN	O	O
.	NN	O	O

While	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
JAK2	NN	O	B-protein
,	NN	O	O
Shc	NN	O	B-protein
,	NN	O	O
Erk	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT5	NN	O	B-protein
proteins	NN	O	I-protein
correlated	NN	O	O
with	NN	O	O
hGM-CSF	NN	O	B-protein
-mediated	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
occurred	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
these	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

gamma-Interferon	NN	O	B-protein
-induced	NN	O	O
resistance	NN	O	O
to	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2	NN	O	O
D3	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
:	NN	O	O
a	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
hypercalcemia	NN	O	O
of	NN	O	O
various	NN	O	O
granulomatoses	NN	O	O
.	NN	O	O

The	NN	O	O
hypercalcemia	NN	O	O
of	NN	O	O
various	NN	O	O
granulomatoses	NN	O	O
is	NN	O	O
caused	NN	O	O
by	NN	O	O
endogenous	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D	NN	O	O
[	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
]	NN	O	O
overproduction	NN	O	O
by	NN	O	O
disease-activated	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
inability	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
to	NN	O	O
suppress	NN	O	O
its	NN	O	O
synthesis	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
contrasts	NN	O	O
with	NN	O	O
the	NN	O	O
tight	NN	O	O
control	NN	O	O
of	NN	O	O
its	NN	O	O
production	NN	O	O
in	NN	O	O
macrophage	NN	O	O
precursors	NN	O	O
,	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
(	NN	O	O
PBM	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
whether	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
resistance	NN	O	O
develops	NN	O	O
as	NN	O	O
PBM	NN	O	B-cell_type
differentiate	NN	O	O
to	NN	O	O
macrophages	NN	O	B-cell_type
or	NN	O	O
with	NN	O	O
macrophage	NN	O	O
activation	NN	O	O
.	NN	O	O

Normal	NN	O	O
human	NN	O	B-cell_type
pulmonary	NN	O	I-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
(	NN	O	O
PAM	NN	O	B-cell_type
)	NN	O	O
are	NN	O	O
less	NN	O	O
sensitive	NN	O	O
to	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
than	NN	O	O
PBM	NN	O	B-cell_type
,	NN	O	O
despite	NN	O	O
similar	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
content	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
both	NN	O	O
PBM	NN	O	B-cell_type
and	NN	O	O
PAM	NN	O	B-cell_type
respond	NN	O	O
to	NN	O	O
exogenous	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
synthesis	NN	O	O
and	NN	O	O
inducing	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
degradation	NN	O	O
through	NN	O	O
enhancement	NN	O	O
of	NN	O	O
24-hydroxylase	NN	O	B-protein
mRNA	NN	O	O
levels	NN	O	O
and	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	B-cell_line
mimics	NN	O	O
PAM	NN	O	B-cell_type
in	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
synthesis	NN	O	O
and	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
exogenous	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
.	NN	O	O

We	NN	O	O
utilized	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
examine	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
with	NN	O	O
macrophage	NN	O	O
activation	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
gamma-interferon	NN	O	B-protein
(	NN	O	O
gamma-IFN	NN	O	B-protein
)	NN	O	O
enhances	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
synthesis	NN	O	O
30-fold	NN	O	O
,	NN	O	O
blocks	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
suppression	NN	O	O
of	NN	O	O
its	NN	O	O
synthesis	NN	O	O
,	NN	O	O
and	NN	O	O
reduces	NN	O	O
by	NN	O	O
42.2	NN	O	O
%	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
induction	NN	O	O
of	NN	O	O
its	NN	O	O
degradation	NN	O	O
.	NN	O	O

The	NN	O	O
antagonistic	NN	O	O
effects	NN	O	O
of	NN	O	O
gamma-IFN	NN	O	B-protein
are	NN	O	O
not	NN	O	O
merely	NN	O	O
restricted	NN	O	O
to	NN	O	O
enzymatic	NN	O	O
activities	NN	O	O
.	NN	O	O

In	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
normal	NN	O	O
PBM	NN	O	B-cell_type
,	NN	O	O
gamma-IFN	NN	O	B-protein
inhibits	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
induction	NN	O	O
of	NN	O	O
24-hydroxylase	NN	O	B-protein
mRNA	NN	O	O
levels	NN	O	O
without	NN	O	O
reducing	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
,	NN	O	O
suggesting	NN	O	O
gamma-IFN	NN	O	B-protein
inhibition	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
transactivating	NN	O	O
function	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
explain	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
overproduction	NN	O	O
in	NN	O	O
granulomatoses	NN	O	O
and	NN	O	O
demonstrate	NN	O	O
potent	NN	O	O
inhibition	NN	O	O
by	NN	O	O
gamma-IFN	NN	O	B-protein
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
action	NN	O	O
in	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
cloning	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
,	NN	O	O
CHEMR1	NN	O	B-DNA
,	NN	O	O
encoding	NN	O	O
a	NN	O	O
chemokine	NN	O	B-protein
receptor	NN	O	I-protein
with	NN	O	O
a	NN	O	O
homology	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-protein
C-C	NN	O	I-protein
chemokine	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
CCR-4	NN	O	B-protein
.	NN	O	O

Chemokines	NN	O	B-protein
refer	NN	O	O
to	NN	O	O
a	NN	O	O
rapidly	NN	O	O
expanding	NN	O	O
family	NN	O	O
of	NN	O	O
small	NN	O	O
cytokines	NN	O	B-protein
whose	NN	O	O
primary	NN	O	O
function	NN	O	O
is	NN	O	O
recruitment	NN	O	O
of	NN	O	O
leukocytes	NN	O	O
to	NN	O	O
inflammatory	NN	O	O
sites	NN	O	O
.	NN	O	O

These	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
seven-transmembrane-domain	NN	O	B-protein
containing	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

A	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
,	NN	O	O
CHEMR1	NN	O	B-DNA
,	NN	O	O
resembling	NN	O	O
the	NN	O	O
typical	NN	O	O
G	NN	O	B-protein
protein-coupled	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
was	NN	O	O
isolated	NN	O	O
from	NN	O	O
a	NN	O	O
mouse	NN	O	B-DNA
cytotoxic	NN	O	I-DNA
T-lymphocyte	NN	O	I-DNA
(	NN	O	I-DNA
CTL	NN	O	I-DNA
)	NN	O	I-DNA
library	NN	O	I-DNA
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
in	NN	O	O
mouse	NN	O	O
cell	NN	O	O
lines	NN	O	O
suggests	NN	O	O
that	NN	O	O
its	NN	O	O
expression	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cells	NN	O	O
,	NN	O	O
including	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
CHEMR1	NN	O	B-DNA
gene	NN	O	I-DNA
Scya3r2	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
single-copy	NN	O	B-DNA
gene	NN	O	I-DNA
whose	NN	O	O
open	NN	O	O
reading	NN	O	O
frame	NN	O	O
may	NN	O	O
be	NN	O	O
in	NN	O	O
a	NN	O	O
single	NN	O	O
exon	NN	O	O
and	NN	O	O
maps	NN	O	O
to	NN	O	O
the	NN	O	O
distal	NN	O	O
region	NN	O	O
of	NN	O	O
mouse	NN	O	B-DNA
Chr	NN	O	I-DNA
9	NN	O	I-DNA
where	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
macrophage	NN	O	I-DNA
inflammatory	NN	O	I-DNA
protein-1alpha	NN	O	I-DNA
(	NN	O	I-DNA
MIP-1alpha	NN	O	I-DNA
)	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
Scya3r	NN	O	B-DNA
and	NN	O	O
two	NN	O	O
related	NN	O	O
C-C	NN	O	B-DNA
chemokine	NN	O	I-DNA
receptor-like	NN	O	I-DNA
genes	NN	O	I-DNA
reside	NN	O	O
.	NN	O	O

Amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
comparison	NN	O	O
shows	NN	O	O
that	NN	O	O
CHEMR1	NN	O	B-DNA
is	NN	O	O
84	NN	O	O
%	NN	O	O
identical	NN	O	O
to	NN	O	O
human	NN	O	B-protein
CCR-4	NN	O	I-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
CHEMR1	NN	O	B-DNA
is	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
mouse	NN	O	B-protein
CCR-4	NN	O	I-protein
.	NN	O	O

Binding	NN	O	O
assays	NN	O	O
using	NN	O	O
125I-labeled	NN	O	B-protein
C-C	NN	O	I-protein
chemokines	NN	O	I-protein
in	NN	O	O
mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
indicated	NN	O	O
that	NN	O	O
CHEMR1	NN	O	B-DNA
did	NN	O	O
not	NN	O	O
bind	NN	O	O
MIP-1alpha	NN	O	O
,	NN	O	O
RANTES	NN	O	O
,	NN	O	O
or	NN	O	O
MIP-1beta	NN	O	O
,	NN	O	O
whereas	NN	O	O
CCR-1	NN	O	B-protein
binds	NN	O	O
MIP-1alpha	NN	O	B-protein
and	NN	O	O
RANTES	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
result	NN	O	O
is	NN	O	O
different	NN	O	O
from	NN	O	O
the	NN	O	O
reported	NN	O	O
properties	NN	O	O
of	NN	O	O
human	NN	O	B-protein
CCR-4	NN	O	I-protein
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
CHEMR1	NN	O	B-DNA
may	NN	O	O
be	NN	O	O
a	NN	O	O
receptor	NN	O	O
for	NN	O	O
unidentified	NN	O	O
C-C	NN	O	B-protein
chemokine	NN	O	I-protein
or	NN	O	O
a	NN	O	O
low-affinity	NN	O	O
receptor	NN	O	O
for	NN	O	O
MIP-1alpha	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Transactivation	NN	O	O
by	NN	O	O
CIITA	NN	O	B-protein
,	NN	O	O
the	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
bare	NN	O	I-protein
lymphocyte	NN	O	I-protein
syndrome-associated	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
requires	NN	O	O
participation	NN	O	O
of	NN	O	O
multiple	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
TATA	NN	O	B-protein
box	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

CIITA	NN	O	B-protein
is	NN	O	O
a	NN	O	O
positive	NN	O	O
regulator	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
major	NN	O	I-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
that	NN	O	O
has	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
defective	NN	O	O
in	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
five	NN	O	O
complementation	NN	O	O
groups	NN	O	O
of	NN	O	O
class	NN	O	B-cell_line
II	NN	O	I-cell_line
major	NN	O	I-cell_line
histocompatibility	NN	O	I-cell_line
complex-negative	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Its	NN	O	O
N-terminal	NN	O	B-protein
region	NN	O	I-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
activating	NN	O	O
transcription	NN	O	O
from	NN	O	O
a	NN	O	O
reporter	NN	O	O
gene	NN	O	O
when	NN	O	O
fused	NN	O	O
to	NN	O	O
a	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
transactivation	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
CIITA	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
by	NN	O	O
studying	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
and	NN	O	O
elongation	NN	O	O
.	NN	O	O

Specifically	NN	O	O
the	NN	O	O
altered	NN	O	O
specificity	NN	O	O
TBP	NN	O	O
(	NN	O	O
TATA	NN	O	B-protein
box	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
assay	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
CIITA	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
to	NN	O	O
mutations	NN	O	O
in	NN	O	O
TBP	NN	O	B-protein
known	NN	O	O
to	NN	O	O
disrupt	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
its	NN	O	O
associated	NN	O	B-protein
general	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Transactivation	NN	O	O
by	NN	O	O
CIITA	NN	O	B-protein
was	NN	O	O
extremely	NN	O	O
sensitive	NN	O	O
to	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
TBP	NN	O	B-protein
that	NN	O	O
in	NN	O	O
yeast	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
abolish	NN	O	O
VP16	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
but	NN	O	O
leaves	NN	O	O
basal	NN	O	O
transcription	NN	O	O
unaffected	NN	O	O
.	NN	O	O

A	NN	O	O
TBP	NN	O	O
mutant	NN	O	O
defective	NN	O	O
in	NN	O	O
interaction	NN	O	O
with	NN	O	O
TBP-associated	NN	O	B-protein
factor	NN	O	I-protein
TAFII250	NN	O	B-protein
also	NN	O	O
failed	NN	O	O
to	NN	O	O
mediate	NN	O	O
transactivation	NN	O	O
through	NN	O	O
the	NN	O	O
CIITA	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Certain	NN	O	O
interactions	NN	O	O
between	NN	O	O
TBP	NN	O	B-protein
and	NN	O	O
general	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
specifically	NN	O	O
required	NN	O	O
for	NN	O	O
acidic	NN	O	B-protein
activation	NN	O	I-protein
domains	NN	O	I-protein
were	NN	O	O
also	NN	O	O
required	NN	O	O
for	NN	O	O
CIITA	NN	O	B-protein
-mediated	NN	O	O
transactivation	NN	O	O
to	NN	O	O
reach	NN	O	O
its	NN	O	O
full	NN	O	O
potential	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
like	NN	O	O
VP16	NN	O	O
,	NN	O	O
CIITA	NN	O	B-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
stimulate	NN	O	O
elongation	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

Overall	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
transactivation	NN	O	O
by	NN	O	O
the	NN	O	O
human	NN	O	B-protein
B-cell-specific	NN	O	I-protein
CIITA	NN	O	I-protein
is	NN	O	O
very	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
herpes	NN	O	B-protein
virus	NN	O	I-protein
transactivator	NN	O	I-protein
VP16	NN	O	B-protein
in	NN	O	O
the	NN	O	O
ways	NN	O	O
that	NN	O	O
have	NN	O	O
been	NN	O	O
tested	NN	O	O
.	NN	O	O

-DOCSTART-	O

Relief	NN	O	O
of	NN	O	O
cyclin	NN	O	B-protein
A	NN	O	I-protein
gene	NN	O	O
transcriptional	NN	O	O
inhibition	NN	O	O
during	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
primary	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
via	NN	O	O
CD2	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

Cyclin	NN	O	B-protein
A	NN	O	I-protein
transcription	NN	O	O
is	NN	O	O
cell	NN	O	O
cycle	NN	O	O
regulated	NN	O	O
and	NN	O	O
induced	NN	O	O
by	NN	O	O
cell	NN	O	O
proliferative	NN	O	O
signals	NN	O	O
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
underlined	NN	O	O
in	NN	O	O
this	NN	O	O
regulation	NN	O	O
in	NN	O	O
normal	NN	O	O
human	NN	O	O
cells	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
analysed	NN	O	O
in	NN	O	O
vivo	NN	O	O
protein-DNA	NN	O	O
interactions	NN	O	O
at	NN	O	O
the	NN	O	O
Cyclin	NN	O	B-protein
A	NN	O	I-protein
locus	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
purified	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
by	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
directed	NN	O	O
at	NN	O	O
CD2	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
gives	NN	O	O
rise	NN	O	O
to	NN	O	O
a	NN	O	O
long	NN	O	O
lasting	NN	O	O
proliferation	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
accessory	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cyclin	NN	O	B-protein
A	NN	O	I-protein
was	NN	O	O
observed	NN	O	O
after	NN	O	O
4	NN	O	O
days	NN	O	O
of	NN	O	O
costimulation	NN	O	O
with	NN	O	O
anti	NN	O	B-protein
CD2	NN	O	I-protein
+	NN	O	O
CD28	NN	O	B-protein
whereas	NN	O	O
stimulation	NN	O	O
by	NN	O	O
anti	NN	O	B-protein
CD2	NN	O	I-protein
or	NN	O	O
anti	NN	O	B-protein
CD28	NN	O	I-protein
alone	NN	O	O
was	NN	O	O
not	NN	O	O
effective	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
genomic	NN	O	O
DMS	NN	O	O
footprinting	NN	O	O
revealed	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
protein	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
two	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
constitutively	NN	O	O
occupied	NN	O	O
.	NN	O	O

They	NN	O	O
bind	NN	O	O
in	NN	O	O
vitro	NN	O	O
respectively	NN	O	O
ATF-1	NN	O	B-protein
and	NN	O	O
NF-Y	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
third	NN	O	O
site	NN	O	O
was	NN	O	O
occupied	NN	O	O
in	NN	O	O
quiescent	NN	O	O
cells	NN	O	O
or	NN	O	O
in	NN	O	O
cells	NN	O	O
stimulated	NN	O	O
by	NN	O	O
anti	NN	O	B-protein
CD2	NN	O	I-protein
or	NN	O	O
anti	NN	O	B-protein
CD28	NN	O	I-protein
alone	NN	O	O
.	NN	O	O

The	NN	O	O
mitogenic	NN	O	O
combination	NN	O	O
of	NN	O	O
anti	NN	O	B-protein
CD2	NN	O	I-protein
+	NN	O	O
anti	NN	O	B-protein
CD28	NN	O	I-protein
released	NN	O	O
the	NN	O	O
footprint	NN	O	O
as	NN	O	O
cells	NN	O	O
were	NN	O	O
committed	NN	O	O
to	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
theses	NN	O	O
results	NN	O	O
,	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
from	NN	O	O
quiescent	NN	O	O
cells	NN	O	O
formed	NN	O	O
a	NN	O	O
specific	NN	O	O
complex	NN	O	O
with	NN	O	O
this	NN	O	O
element	NN	O	O
,	NN	O	O
whereas	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
from	NN	O	O
cells	NN	O	O
treated	NN	O	O
with	NN	O	O
anti	NN	O	B-protein
CD2	NN	O	I-protein
+	NN	O	O
anti	NN	O	B-protein
CD28	NN	O	I-protein
failed	NN	O	O
to	NN	O	O
do	NN	O	O
so	NN	O	O
after	NN	O	O
cells	NN	O	O
entered	NN	O	O
a	NN	O	O
proliferative	NN	O	O
state	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
enhancer-blocking	NN	O	B-DNA
element	NN	O	I-DNA
between	NN	O	O
alpha	NN	O	B-DNA
and	NN	O	I-DNA
delta	NN	O	I-DNA
gene	NN	O	I-DNA
segments	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha/delta	NN	O	I-DNA
locus	NN	O	I-DNA
.	NN	O	O

T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
TCR	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
delta	NN	O	O
gene	NN	O	O
segments	NN	O	O
are	NN	O	O
organized	NN	O	O
within	NN	O	O
a	NN	O	O
single	NN	O	O
genetic	NN	O	O
locus	NN	O	O
but	NN	O	O
are	NN	O	O
differentially	NN	O	O
regulated	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

An	NN	O	O
enhancer-blocking	NN	O	B-DNA
element	NN	O	I-DNA
(	NN	O	O
BEAD-1	NN	O	B-DNA
,	NN	O	O
for	NN	O	O
blocking	NN	O	O
element	NN	O	B-DNA
alpha/delta	NN	O	I-DNA
1	NN	O	I-DNA
)	NN	O	O
was	NN	O	O
localized	NN	O	O
to	NN	O	O
a	NN	O	O
2.0-kb	NN	O	B-DNA
region	NN	O	I-DNA
3	NN	O	O
'	NN	O	O
of	NN	O	O
TCR	NN	O	B-DNA
delta	NN	O	I-DNA
gene	NN	O	I-DNA
segments	NN	O	I-DNA
and	NN	O	O
5	NN	O	O
'	NN	O	O
of	NN	O	O
TCR	NN	O	B-DNA
alpha	NN	O	I-DNA
joining	NN	O	I-DNA
gene	NN	O	I-DNA
segments	NN	O	I-DNA
within	NN	O	O
this	NN	O	O
locus	NN	O	O
.	NN	O	O

BEAD-1	NN	O	B-DNA
blocked	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
delta	NN	O	I-DNA
enhancer	NN	O	I-DNA
(	NN	O	O
Edelta	NN	O	B-DNA
)	NN	O	O
to	NN	O	O
activate	NN	O	O
a	NN	O	O
promoter	NN	O	O
when	NN	O	O
located	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
in	NN	O	O
a	NN	O	O
chromatin-integrated	NN	O	B-DNA
construct	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
BEAD-1	NN	O	B-DNA
functions	NN	O	O
as	NN	O	O
a	NN	O	O
boundary	NN	O	O
that	NN	O	O
separates	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
alpha/delta	NN	O	I-DNA
locus	NN	O	I-DNA
into	NN	O	O
distinct	NN	O	O
regulatory	NN	O	B-DNA
domains	NN	O	I-DNA
controlled	NN	O	O
by	NN	O	O
Edelta	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
alpha	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
that	NN	O	O
it	NN	O	O
prevents	NN	O	O
Edelta	NN	O	B-DNA
from	NN	O	O
opening	NN	O	O
the	NN	O	O
chromatin	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
alpha	NN	O	I-DNA
joining	NN	O	O
gene	NN	O	O
segments	NN	O	O
for	NN	O	O
VDJ	NN	O	O
recombination	NN	O	O
at	NN	O	O
an	NN	O	O
early	NN	O	O
stage	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Uncoupling	NN	O	O
of	NN	O	O
cell	NN	O	O
cycle	NN	O	O
arrest	NN	O	O
from	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
markers	NN	O	O
in	NN	O	O
HL60	NN	O	B-cell_line
cell	NN	O	I-cell_line
variants	NN	O	I-cell_line
.	NN	O	O

Differentiation	NN	O	O
generally	NN	O	O
leads	NN	O	O
to	NN	O	O
cell	NN	O	O
cycle	NN	O	O
arrest	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
HL60	NN	O	I-cell_line
cells	NN	O	I-cell_line
respond	NN	O	O
to	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
1	NN	O	O
,	NN	O	O
25D3	NN	O	O
)	NN	O	O
by	NN	O	O
expressing	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
markers	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	O
phenotype	NN	O	O
and	NN	O	O
become	NN	O	O
arrested	NN	O	O
predominantly	NN	O	O
in	NN	O	O
the	NN	O	O
G1	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
reported	NN	O	O
a	NN	O	O
series	NN	O	O
(	NN	O	O
A	NN	O	O
)	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25D3-resistant	NN	O	O
variants	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
which	NN	O	O
proliferate	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25D3	NN	O	O
and	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
differentiation	NN	O	O
markers	NN	O	O
(	NN	O	O
Exp.	NN	O	O
Cell	NN	O	O
Res.	NN	O	O
224	NN	O	O
,	NN	O	O
312	NN	O	O
,	NN	O	O
1996	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
describe	NN	O	O
another	NN	O	O
series	NN	O	O
(	NN	O	O
B	NN	O	O
)	NN	O	O
of	NN	O	O
such	NN	O	O
variants	NN	O	O
,	NN	O	O
which	NN	O	O
differ	NN	O	O
from	NN	O	O
A	NN	O	O
series	NN	O	O
cells	NN	O	O
grown	NN	O	O
in	NN	O	O
similar	NN	O	O
concentrations	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25D3	NN	O	O
in	NN	O	O
that	NN	O	O
they	NN	O	O
express	NN	O	O
the	NN	O	O
CD14	NN	O	B-protein
antigen	NN	O	I-protein
and	NN	O	O
nonspecific	NN	O	O
esterase	NN	O	B-protein
,	NN	O	O
characteristic	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte	NN	O	O
,	NN	O	O
while	NN	O	O
continuing	NN	O	O
to	NN	O	O
proliferate	NN	O	O
and	NN	O	O
they	NN	O	O
develop	NN	O	O
hypotetraploid	NN	O	O
DNA	NN	O	O
(	NN	O	O
4C	NN	O	O
)	NN	O	O
content	NN	O	O
at	NN	O	O
higher	NN	O	O
concentrations	NN	O	O
of	NN	O	O
ambient	NN	O	O
1	NN	O	O
,	NN	O	O
25D3	NN	O	O
than	NN	O	O
the	NN	O	O
A	NN	O	O
series	NN	O	O
cells	NN	O	O
.	NN	O	O

Cells	NN	O	O
in	NN	O	O
the	NN	O	O
B	NN	O	O
series	NN	O	O
with	NN	O	O
4C	NN	O	O
DNA	NN	O	O
content	NN	O	O
(	NN	O	O
100B	NN	O	O
and	NN	O	O
200B	NN	O	O
)	NN	O	O
also	NN	O	O
differed	NN	O	O
from	NN	O	O
the	NN	O	O
A	NN	O	B-cell_line
series	NN	O	I-cell_line
4C	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
the	NN	O	O
full-length	NN	O	O
Sp1	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
B	NN	O	O
series	NN	O	O
cells	NN	O	O
resembled	NN	O	O
the	NN	O	O
A	NN	O	O
series	NN	O	O
cells	NN	O	O
in	NN	O	O
exhibiting	NN	O	O
faster	NN	O	O
growth	NN	O	O
rates	NN	O	O
than	NN	O	O
the	NN	O	O
parental	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
showed	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
retinoid	NN	O	B-protein
receptor	NN	O	I-protein
X	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
initial	NN	O	O
steps	NN	O	O
in	NN	O	O
the	NN	O	O
1	NN	O	O
,	NN	O	O
25D3	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
are	NN	O	O
intact	NN	O	O
in	NN	O	O
B	NN	O	O
series	NN	O	O
resistant	NN	O	O
cells	NN	O	O
and	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
early	NN	O	O
markers	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
progression	NN	O	O
to	NN	O	O
subsequent	NN	O	O
events	NN	O	O
which	NN	O	O
comprise	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
arrest	NN	O	O
is	NN	O	O
halted	NN	O	O
during	NN	O	O
the	NN	O	O
adaptation	NN	O	O
to	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25D3	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
availability	NN	O	O
of	NN	O	O
these	NN	O	O
variant	NN	O	O
cells	NN	O	O
should	NN	O	O
provide	NN	O	O
a	NN	O	O
system	NN	O	O
for	NN	O	O
studying	NN	O	O
the	NN	O	O
link	NN	O	O
between	NN	O	O
differentiation	NN	O	O
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
arrest	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
replication	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
a	NN	O	O
novel	NN	O	O
combination	NN	O	O
of	NN	O	O
anti-Tat	NN	O	B-protein
single-chain	NN	O	I-protein
intrabodies	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
antagonists	NN	O	O
.	NN	O	O

Human	NN	O	B-protein
immunodeficiency	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
HIV-1	NN	O	I-protein
)	NN	O	I-protein
Tat	NN	O	I-protein
,	NN	O	O
an	NN	O	O
early	NN	O	O
regulatory	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
replication	NN	O	O
,	NN	O	O
transactivates	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
via	NN	O	O
its	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
transactivation	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
TAR	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
,	NN	O	O
along	NN	O	O
with	NN	O	O
other	NN	O	O
cellular	NN	O	O
factors	NN	O	O
,	NN	O	O
increases	NN	O	O
viral	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
and	NN	O	O
elongation	NN	O	O
.	NN	O	O

Tat	NN	O	O
also	NN	O	O
superactivates	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
promoter	NN	O	I-DNA
through	NN	O	O
a	NN	O	O
TAR	NN	O	B-DNA
-independent	NN	O	O
mechanism	NN	O	O
,	NN	O	O
including	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor	NN	O	O
alpha-induced	NN	O	O
and	NN	O	O
protein	NN	O	O
kinase	NN	O	O
C	NN	O	O
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
Tat	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
cooperatively	NN	O	O
down-regulate	NN	O	O
this	NN	O	O
Tat	NN	O	B-protein
-mediated	NN	O	O
LTR	NN	O	B-DNA
superactivation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
a	NN	O	O
combined	NN	O	O
pharmacologic	NN	O	O
and	NN	O	O
genetic	NN	O	O
strategy	NN	O	O
using	NN	O	O
two	NN	O	O
PKC	NN	O	B-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
pentoxifylline	NN	O	O
(	NN	O	O
PTX	NN	O	O
)	NN	O	O
and	NN	O	O
Go-6976	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
stably	NN	O	O
expressed	NN	O	O
anti-Tat	NN	O	B-protein
single-chain	NN	O	I-protein
intracellular	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
sFv	NN	O	B-protein
intrabody	NN	O	I-protein
)	NN	O	O
was	NN	O	O
employed	NN	O	O
to	NN	O	O
obtain	NN	O	O
cooperative	NN	O	O
inhibition	NN	O	O
of	NN	O	O
both	NN	O	O
HIV-1	NN	O	O
LTR	NN	O	B-DNA
-driven	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
PTX	NN	O	O
and	NN	O	O
Go-6976	NN	O	O
resulted	NN	O	O
in	NN	O	O
cooperative	NN	O	O
inhibition	NN	O	O
of	NN	O	O
both	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
-driven	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
combined	NN	O	O
use	NN	O	O
of	NN	O	O
anti-Tat	NN	O	B-protein
sFv	NN	O	I-protein
intrabodies	NN	O	I-protein
and	NN	O	O
the	NN	O	O
two	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibitors	NN	O	O
retained	NN	O	O
the	NN	O	O
virus	NN	O	O
in	NN	O	O
the	NN	O	O
latent	NN	O	O
state	NN	O	O
for	NN	O	O
as	NN	O	O
long	NN	O	O
as	NN	O	O
45	NN	O	O
days	NN	O	O
.	NN	O	O

The	NN	O	O
combined	NN	O	O
treatment	NN	O	O
resulted	NN	O	O
in	NN	O	O
more	NN	O	O
durable	NN	O	O
inhibition	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
than	NN	O	O
was	NN	O	O
seen	NN	O	O
with	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibitors	NN	O	O
alone	NN	O	O
or	NN	O	O
the	NN	O	O
anti-	NN	O	O
Tat	NN	O	B-protein
sFv	NN	O	I-protein
intrabodies	NN	O	I-protein
alone	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
in	NN	O	O
future	NN	O	O
clinical	NN	O	O
gene	NN	O	O
therapy	NN	O	O
trials	NN	O	O
,	NN	O	O
a	NN	O	O
combined	NN	O	O
pharmacologic	NN	O	O
and	NN	O	O
genetic	NN	O	O
strategy	NN	O	O
like	NN	O	O
the	NN	O	O
one	NN	O	O
reported	NN	O	O
here	NN	O	O
may	NN	O	O
improve	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
transduced	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
prolong	NN	O	O
clinical	NN	O	O
benefit	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
immune-mediated	NN	O	O
disease	NN	O	O
.	NN	O	O

A	NN	O	O
large	NN	O	O
amount	NN	O	O
of	NN	O	O
detailed	NN	O	O
information	NN	O	O
about	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
proteins	NN	O	I-protein
regulating	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
the	NN	O	O
cellular	NN	O	O
response	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
has	NN	O	O
emerged	NN	O	O
recently	NN	O	O
.	NN	O	O

Several	NN	O	O
small	NN	O	O
molecules	NN	O	O
,	NN	O	O
an	NN	O	O
antisense	NN	O	B-DNA
oligonucleotide	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
gene	NN	O	O
therapeutic	NN	O	O
agents	NN	O	O
that	NN	O	O
inhibit	NN	O	O
NF-kappa	NN	O	B-protein
b	NN	O	I-protein
activation	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

Despite	NN	O	O
this	NN	O	O
,	NN	O	O
there	NN	O	O
are	NN	O	O
still	NN	O	O
significant	NN	O	O
gaps	NN	O	O
in	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
this	NN	O	O
process	NN	O	O
and	NN	O	O
its	NN	O	O
consequences	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
selectively	NN	O	O
regulating	NN	O	O
cytokine	NN	O	O
production	NN	O	O
by	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
subsets	NN	O	I-cell_type
is	NN	O	O
at	NN	O	O
a	NN	O	O
very	NN	O	O
early	NN	O	O
stage	NN	O	O
.	NN	O	O

Three	NN	O	O
interacting	NN	O	B-protein
proteins	NN	O	I-protein
have	NN	O	O
recently	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
contribute	NN	O	O
to	NN	O	O
subset-restricted	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

There	NN	O	O
are	NN	O	O
other	NN	O	O
elements	NN	O	O
regulating	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
relative	NN	O	O
importance	NN	O	O
of	NN	O	O
these	NN	O	O
recently	NN	O	O
identified	NN	O	O
proteins	NN	O	O
has	NN	O	O
yet	NN	O	O
to	NN	O	O
be	NN	O	O
determined	NN	O	O
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
Rel	NN	O	B-protein
,	NN	O	I-protein
Fos	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
Jun	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
binding	NN	O	O
activity	NN	O	O
to	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
CD28	NN	O	I-DNA
response	NN	O	I-DNA
element/AP-1	NN	O	I-DNA
sequence	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CD28	NN	O	B-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
costimulatory	NN	O	B-protein
molecule	NN	O	I-protein
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Costimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
both	NN	O	O
the	NN	O	O
Ag	NN	O	O
receptor	NN	O	O
and	NN	O	O
CD28	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
high	NN	O	O
level	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
vital	NN	O	O
to	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
an	NN	O	O
immune	NN	O	O
response	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Previous	NN	O	O
reports	NN	O	O
have	NN	O	O
suggested	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
stimulation	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
up-regulating	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	I-protein
NF-kappaB	NN	O	I-protein
)	NN	O	I-protein
/Rel	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
an	NN	O	O
uncharacterized	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
called	NN	O	O
CD28	NN	O	B-protein
response	NN	O	O
complex	NN	O	O
.	NN	O	O

While	NN	O	O
several	NN	O	O
lines	NN	O	O
of	NN	O	O
investigation	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
make	NN	O	O
up	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
response	NN	O	O
complex	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
other	NN	O	O
work	NN	O	O
has	NN	O	O
not	NN	O	O
supported	NN	O	O
this	NN	O	O
conclusion	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
CD28	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CD28RE	NN	O	B-DNA
)	NN	O	O
does	NN	O	O
not	NN	O	O
function	NN	O	O
independently	NN	O	O
but	NN	O	O
works	NN	O	O
instead	NN	O	O
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
the	NN	O	O
adjacent	NN	O	B-DNA
promoter	NN	O	I-DNA
proximal	NN	O	I-DNA
AP-1-binding	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
is	NN	O	O
confirmed	NN	O	O
here	NN	O	O
.	NN	O	O

Also	NN	O	O
in	NN	O	O
the	NN	O	O
current	NN	O	O
study	NN	O	O
,	NN	O	O
binding	NN	O	O
activity	NN	O	O
to	NN	O	O
the	NN	O	O
CD28RE/AP-1	NN	O	B-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
evaluated	NN	O	O
.	NN	O	O

Although	NN	O	O
four	NN	O	O
specific	NN	O	O
complexes	NN	O	O
can	NN	O	O
be	NN	O	O
detected	NN	O	O
binding	NN	O	O
to	NN	O	O
this	NN	O	O
sequence	NN	O	O
,	NN	O	O
only	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
complexes	NN	O	O
is	NN	O	O
specific	NN	O	O
for	NN	O	O
both	NN	O	O
the	NN	O	O
CD28RE	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
adjacent	NN	O	B-DNA
AP-1	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Of	NN	O	O
the	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
tested	NN	O	O
,	NN	O	O
this	NN	O	O
CD28RE/AP-1-specific	NN	O	B-protein
complex	NN	O	I-protein
contains	NN	O	O
predominantly	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
despite	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
both	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
RelA	NN	O	B-protein
can	NN	O	O
efficiently	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
CD28RE	NN	O	B-DNA
.	NN	O	O

c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
are	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
this	NN	O	O
CD28RE/AP-1-specific	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
functional	NN	O	O
complexes	NN	O	O
encompassing	NN	O	O
both	NN	O	O
the	NN	O	O
CD28RE	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
AP-1-binding	NN	O	B-DNA
sites	NN	O	I-DNA
influence	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
CD28	NN	O	B-protein
-costimulated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Inducible	NN	O	O
expression	NN	O	O
and	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
coactivator	NN	O	B-protein
BOB.1/OBF.1	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

BOB.1/OBF.1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
coactivator	NN	O	I-protein
that	NN	O	O
is	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
Oct1	NN	O	O
and	NN	O	O
Oct2	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Upon	NN	O	O
activation	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
primary	NN	O	B-cell_type
murine	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
,	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
transactivation	NN	O	O
function	NN	O	O
were	NN	O	O
induced	NN	O	O
.	NN	O	O

BOB.1/OBF.1	NN	O	B-protein
was	NN	O	O
phosphorylated	NN	O	O
at	NN	O	O
Ser184	NN	O	O
both	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
modification	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
inducible	NN	O	O
activation	NN	O	O
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
Ser184	NN	O	O
also	NN	O	O
diminished	NN	O	O
transactivation	NN	O	O
function	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
activating	NN	O	O
phosphorylation	NN	O	O
that	NN	O	O
is	NN	O	O
inducible	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
constitutively	NN	O	O
present	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
coactivator	NN	O	I-protein
that	NN	O	O
is	NN	O	O
critically	NN	O	O
regulated	NN	O	O
by	NN	O	O
posttranslational	NN	O	O
modifications	NN	O	O
to	NN	O	O
mediate	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Increased	NN	O	O
expression	NN	O	O
of	NN	O	O
Gs	NN	O	B-protein
(	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
enhances	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
adenylyl	NN	O	B-protein
cyclase	NN	O	I-protein
signal	NN	O	O
transduction	NN	O	O
cascade	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
stimulatory	NN	O	O
G	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
can	NN	O	O
vary	NN	O	O
over	NN	O	O
a	NN	O	O
3-fold	NN	O	O
range	NN	O	O
in	NN	O	O
human	NN	O	O
tissues	NN	O	O
and	NN	O	O
in	NN	O	O
rodent	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
the	NN	O	O
offspring	NN	O	O
of	NN	O	O
alcoholics	NN	O	O
have	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
certain	NN	O	O
tissues	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
offspring	NN	O	O
of	NN	O	O
nonalcoholics	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
research	NN	O	O
was	NN	O	O
to	NN	O	O
test	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
a	NN	O	O
causal	NN	O	O
relationship	NN	O	O
exists	NN	O	O
between	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
adenylyl	NN	O	B-protein
cyclase	NN	O	I-protein
(	NN	O	O
AC	NN	O	O
)	NN	O	O
cascade	NN	O	O
.	NN	O	O

The	NN	O	O
methodology	NN	O	O
employed	NN	O	O
transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
HEK	NN	O	B-cell_line
293	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
a	NN	O	O
cDNA	NN	O	O
for	NN	O	O
the	NN	O	O
52-kDa	NN	O	O
form	NN	O	O
of	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
under	NN	O	O
regulation	NN	O	O
by	NN	O	O
inducible	NN	O	B-DNA
metallothionein	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

Transfectants	NN	O	O
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
varying	NN	O	O
concentrations	NN	O	O
(	NN	O	O
0-125	NN	O	O
microM	NN	O	O
)	NN	O	O
of	NN	O	O
zinc	NN	O	O
sulfate	NN	O	O
that	NN	O	O
produced	NN	O	O
a	NN	O	O
3-fold	NN	O	O
range	NN	O	O
of	NN	O	O
membrane	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
range	NN	O	O
of	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
expression	NN	O	O
produced	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
mimic	NN	O	O
a	NN	O	O
physiologically	NN	O	O
relevant	NN	O	O
spectrum	NN	O	O
of	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
membranes	NN	O	O
derived	NN	O	O
from	NN	O	O
human	NN	O	O
tissues	NN	O	O
and	NN	O	O
rat	NN	O	O
brain	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
observed	NN	O	O
that	NN	O	O
induction	NN	O	O
of	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
expression	NN	O	O
increased	NN	O	O
constitutive	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
stimulated	NN	O	O
cAMP	NN	O	O
accumulation	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
expression	NN	O	O
increased	NN	O	O
events	NN	O	O
distal	NN	O	O
to	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
cAMP	NN	O	O
including	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
,	NN	O	O
cAMP	NN	O	B-protein
response	NN	O	I-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
cAMP-dependent	NN	O	B-DNA
reporter	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
these	NN	O	O
studies	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
expression	NN	O	O
has	NN	O	O
a	NN	O	O
marked	NN	O	O
impact	NN	O	O
on	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
AC	NN	O	O
cascade	NN	O	O
from	NN	O	O
the	NN	O	O
membrane	NN	O	O
through	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
individuals	NN	O	O
who	NN	O	O
differ	NN	O	O
in	NN	O	O
G	NN	O	B-protein
(	NN	O	I-protein
S	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
expression	NN	O	O
may	NN	O	O
also	NN	O	O
differ	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
certain	NN	O	O
cAMP-dependent	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Thrombin	NN	O	B-protein
generation	NN	O	O
by	NN	O	O
apoptotic	NN	O	B-cell_type
vascular	NN	O	I-cell_type
smooth	NN	O	I-cell_type
muscle	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Thrombin	NN	O	B-protein
activation	NN	O	O
requires	NN	O	O
assembly	NN	O	O
of	NN	O	O
a	NN	O	O
prothrombinase	NN	O	B-protein
complex	NN	O	I-protein
of	NN	O	O
activated	NN	O	O
coagulation	NN	O	B-protein
factors	NN	O	I-protein
on	NN	O	O
an	NN	O	O
anionic	NN	O	O
phospholipid	NN	O	O
surface	NN	O	O
,	NN	O	O
classically	NN	O	O
provided	NN	O	O
by	NN	O	O
activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
anionic	NN	O	O
phosphatidylserine	NN	O	O
is	NN	O	O
exposed	NN	O	O
by	NN	O	O
rat	NN	O	B-cell_type
vascular	NN	O	I-cell_type
smooth	NN	O	I-cell_type
muscle	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
VSMCs	NN	O	B-cell_type
)	NN	O	O
undergoing	NN	O	O
apoptosis	NN	O	O
after	NN	O	O
serum	NN	O	O
withdrawal	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
chromogenic	NN	O	O
assay	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
thrombin	NN	O	B-protein
generation	NN	O	O
by	NN	O	O
apoptotic	NN	O	B-cell_type
VSMCs	NN	O	I-cell_type
expressing	NN	O	O
c-myc	NN	O	B-protein
(	NN	O	O
VSMC-myc	NN	O	B-cell_line
)	NN	O	O
with	NN	O	O
an	NN	O	O
area	NN	O	O
under	NN	O	O
the	NN	O	O
thrombin	NN	O	B-protein
-generation	NN	O	O
curve	NN	O	O
(	NN	O	O
AUC	NN	O	O
)	NN	O	O
of	NN	O	O
305	NN	O	O
+/-	NN	O	O
17	NN	O	O
nmol	NN	O	O
x	NN	O	O
min/L	NN	O	O
and	NN	O	O
a	NN	O	O
peak	NN	O	O
thrombin	NN	O	B-protein
(	NN	O	O
PT	NN	O	O
)	NN	O	O
of	NN	O	O
154	NN	O	O
+/-	NN	O	O
9	NN	O	O
nmol/L	NN	O	O
.	NN	O	O

The	NN	O	O
thrombin	NN	O	B-protein
-generating	NN	O	O
potential	NN	O	O
of	NN	O	O
the	NN	O	O
apoptotic	NN	O	B-cell_line
VSMC-myc	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
greater	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
unactivated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
(	NN	O	O
P	NN	O	O
=	NN	O	O
.003	NN	O	O
for	NN	O	O
AUC	NN	O	O
;	NN	O	O
P	NN	O	O
=	NN	O	O
.0002	NN	O	O
for	NN	O	O
PT	NN	O	O
)	NN	O	O
and	NN	O	O
similar	NN	O	O
to	NN	O	O
calcium-ionophore	NN	O	O
activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
(	NN	O	O
AUC	NN	O	O
of	NN	O	O
332	NN	O	O
+/-	NN	O	O
15	NN	O	O
nmol	NN	O	O
x	NN	O	O
min/L	NN	O	O
,	NN	O	O
P	NN	O	O
=	NN	O	O
.3	NN	O	O
;	NN	O	O
PT	NN	O	O
of	NN	O	O
172	NN	O	O
+/-	NN	O	O
8	NN	O	O
nmol/L	NN	O	O
,	NN	O	O
P	NN	O	O
=	NN	O	O
.2	NN	O	O
)	NN	O	O
.	NN	O	O

Thrombin	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
also	NN	O	O
seen	NN	O	O
with	NN	O	O
apoptotic	NN	O	B-cell_type
human	NN	O	I-cell_type
VSMCs	NN	O	I-cell_type
(	NN	O	O
AUC	NN	O	O
of	NN	O	O
211	NN	O	O
+/-	NN	O	O
8	NN	O	O
nmol	NN	O	O
x	NN	O	O
min/L	NN	O	O
;	NN	O	O
PT	NN	O	O
of	NN	O	O
103	NN	O	O
+/-	NN	O	O
4	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
and	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
annexin	NN	O	O
V	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
.0001	NN	O	O
for	NN	O	O
AUC	NN	O	O
and	NN	O	O
PT	NN	O	O
)	NN	O	O
.	NN	O	O

VSMC-myc	NN	O	B-cell_line
cells	NN	O	I-cell_line
maintained	NN	O	O
in	NN	O	O
serum	NN	O	O
generated	NN	O	O
less	NN	O	O
thrombin	NN	O	B-protein
than	NN	O	O
after	NN	O	O
serum	NN	O	O
withdrawal	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.0002	NN	O	O
for	NN	O	O
AUC	NN	O	O
and	NN	O	O
PT	NN	O	O
)	NN	O	O
.	NN	O	O

VSMCs	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
human	NN	O	O
coronary	NN	O	O
atherosclerotic	NN	O	O
plaques	NN	O	O
that	NN	O	O
apoptose	NN	O	O
even	NN	O	O
in	NN	O	O
serum	NN	O	O
also	NN	O	O
generated	NN	O	O
thrombin	NN	O	B-protein
(	NN	O	O
AUC	NN	O	O
of	NN	O	O
260	NN	O	O
+/-	NN	O	O
2	NN	O	O
nmol	NN	O	O
x	NN	O	O
min/L	NN	O	O
;	NN	O	O
PT	NN	O	O
of	NN	O	O
128	NN	O	O
+/-	NN	O	O
4	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
apoptotic	NN	O	B-cell_type
VSMCs	NN	O	I-cell_type
possess	NN	O	O
a	NN	O	O
significant	NN	O	O
thrombin	NN	O	B-protein
-generating	NN	O	O
capacity	NN	O	O
secondary	NN	O	O
to	NN	O	O
phosphatidylserine	NN	O	O
exposure	NN	O	O
.	NN	O	O

Apoptotic	NN	O	O
cells	NN	O	O
within	NN	O	O
atherosclerotic	NN	O	O
plaques	NN	O	O
may	NN	O	O
allow	NN	O	O
local	NN	O	O
thrombin	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
thereby	NN	O	O
contributing	NN	O	O
to	NN	O	O
disease	NN	O	O
progression	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
isotype-specific	NN	O	O
activator	NN	O	O
of	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
(	NN	O	I-DNA
MHC	NN	O	I-DNA
)	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
one	NN	O	O
type	NN	O	O
of	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
combined	NN	O	O
immunodeficiency	NN	O	O
have	NN	O	O
mutations	NN	O	O
in	NN	O	O
a	NN	O	O
gene	NN	O	O
termed	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
coordinately	NN	O	O
controls	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
major	NN	O	O
human	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
HLA-DR	NN	O	B-DNA
,	NN	O	I-DNA
-DQ	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
-DP	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
experimentally	NN	O	O
derived	NN	O	O
B-lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
clone	NN	O	B-cell_line
13	NN	O	I-cell_line
,	NN	O	O
expresses	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
HLADQ	NN	O	B-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-DNA
and	NN	O	O
HLA-DP	NN	O	B-DNA
,	NN	O	O
despite	NN	O	O
its	NN	O	O
mapping	NN	O	O
by	NN	O	O
complementation	NN	O	O
analysis	NN	O	O
to	NN	O	O
this	NN	O	O
group	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
possible	NN	O	O
that	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
clone	NN	O	B-DNA
13	NN	O	I-DNA
CIITA	NN	O	I-DNA
alleles	NN	O	I-DNA
bore	NN	O	O
a	NN	O	O
mutation	NN	O	O
that	NN	O	O
allowed	NN	O	O
HLA-DQ	NN	O	B-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
HLA-DR	NN	O	O
or	NN	O	O
-DP	NN	O	O
transcription	NN	O	O
.	NN	O	O

Alternatively	NN	O	O
,	NN	O	O
another	NN	O	O
factor	NN	O	O
,	NN	O	O
distinct	NN	O	O
from	NN	O	O
CIITA	NN	O	B-protein
,	NN	O	O
might	NN	O	O
control	NN	O	O
HLA-DQ	NN	O	B-DNA
expression	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
CIITA	NN	O	B-DNA
cDNAs	NN	O	I-DNA
derived	NN	O	O
by	NN	O	O
reverse	NN	O	O
transcriptase	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
from	NN	O	O
clone	NN	O	B-cell_line
13	NN	O	I-cell_line
do	NN	O	O
not	NN	O	O
restore	NN	O	O
expression	NN	O	O
of	NN	O	O
HLA-DQ	NN	O	B-DNA
in	NN	O	O
another	NN	O	O
CIITA-deficient	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
RJ2.2.5	NN	O	B-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
no	NN	O	O
CIITA	NN	O	B-protein
protein	NN	O	O
is	NN	O	O
detectable	NN	O	O
in	NN	O	O
clone	NN	O	B-cell_line
13	NN	O	I-cell_line
nuclear	NN	O	O
extracts	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
somatic	NN	O	O
cell	NN	O	O
fusion	NN	O	O
between	NN	O	O
clone	NN	O	B-cell_line
13	NN	O	I-cell_line
and	NN	O	O
RJ2.2.5	NN	O	B-cell_line
restored	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
HLA-DQ	NN	O	B-DNA
haplotype	NN	O	O
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
RJ2.2.5	NN	O	B-DNA
DQB	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
an	NN	O	O
HLA-DQ	NN	O	B-protein
isotype-specific	NN	O	I-protein
trans-acting	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
functions	NN	O	O
independently	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Homodimerization	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
interleukin	NN	O	I-protein
4	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
induces	NN	O	O
Cepsilon	NN	O	B-RNA
germline	NN	O	I-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
receptor	NN	O	I-protein
gamma	NN	O	I-protein
chain	NN	O	I-protein
.	NN	O	O

The	NN	O	O
cytokines	NN	O	B-protein
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-4	NN	O	I-protein
and	NN	O	O
IL-13	NN	O	B-protein
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
inducing	NN	O	O
Cepsilon	NN	O	B-RNA
germline	NN	O	I-RNA
transcripts	NN	O	I-RNA
and	NN	O	O
IgE	NN	O	B-protein
isotype	NN	O	I-protein
switching	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
IL-4	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-4R	NN	O	B-protein
)	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
composed	NN	O	O
of	NN	O	O
two	NN	O	O
chains	NN	O	O
,	NN	O	O
the	NN	O	O
IL-4-binding	NN	O	B-protein
IL-4Ralpha	NN	O	I-protein
chain	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
shared	NN	O	O
with	NN	O	O
the	NN	O	O
IL-13R	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
IL-2Rgamma	NN	O	B-protein
(	NN	O	I-protein
gammac	NN	O	I-protein
)	NN	O	I-protein
chain	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
shared	NN	O	O
with	NN	O	O
IL-7R	NN	O	B-protein
,	NN	O	O
IL-9R	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-15R	NN	O	B-protein
.	NN	O	O

IL-4	NN	O	B-protein
induces	NN	O	O
Cepsilon	NN	O	B-RNA
germline	NN	O	I-RNA
transcripts	NN	O	I-RNA
and	NN	O	O
IgE	NN	O	B-protein
isotype	NN	O	I-protein
switching	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
gammac	NN	O	B-protein
chain	NN	O	I-protein
deficiency	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
Cepsilon	NN	O	B-RNA
germline	NN	O	I-RNA
transcripts	NN	O	I-RNA
by	NN	O	O
IL-4	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
lack	NN	O	O
the	NN	O	O
gammac	NN	O	B-protein
chain	NN	O	I-protein
may	NN	O	O
involve	NN	O	O
signaling	NN	O	O
via	NN	O	O
the	NN	O	O
IL-13R	NN	O	B-protein
.	NN	O	O

Alternatively	NN	O	O
,	NN	O	O
the	NN	O	O
IL-4Ralpha	NN	O	B-protein
chain	NN	O	I-protein
may	NN	O	O
transduce	NN	O	O
intracellular	NN	O	O
signals	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
Cepsilon	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
independently	NN	O	O
of	NN	O	O
its	NN	O	O
association	NN	O	O
with	NN	O	O
other	NN	O	O
chains	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
ligand-induced	NN	O	O
homodimerization	NN	O	O
of	NN	O	O
chimeric	NN	O	B-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
consisting	NN	O	O
of	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
and	NN	O	I-protein
transmembrane	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
the	NN	O	O
erythropoietin	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
of	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
IL-4Ralpha	NN	O	B-protein
induces	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
Jak1	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
,	NN	O	O
STAT6	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
and	NN	O	O
Cepsilon	NN	O	B-RNA
germline	NN	O	I-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
BJAB	NN	O	I-cell_line
.	NN	O	O

Disruption	NN	O	O
of	NN	O	O
the	NN	O	O
Jak1-binding	NN	O	B-protein
proline-rich	NN	O	I-protein
Box1	NN	O	I-protein
region	NN	O	I-protein
of	NN	O	O
IL-4Ralpha	NN	O	B-protein
abolished	NN	O	O
signaling	NN	O	O
by	NN	O	O
this	NN	O	O
chimeric	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
chimeric	NN	O	B-protein
CD8alpha/IL-4Ralpha	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
expressed	NN	O	O
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
as	NN	O	O
a	NN	O	O
homodimer	NN	O	B-protein
,	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
Cepsilon	NN	O	B-RNA
germline	NN	O	I-RNA
transcripts	NN	O	I-RNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
homodimerization	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4Ralpha	NN	O	B-protein
chain	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
transduce	NN	O	O
Jak1	NN	O	B-protein
-dependent	NN	O	O
intracellular	NN	O	O
signals	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
IgE	NN	O	B-protein
isotype	NN	O	I-protein
switching	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
activity	NN	O	O
and	NN	O	O
constitutive	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
the	NN	O	O
ETS	NN	O	B-protein
protein	NN	O	I-protein
Elf-1	NN	O	B-protein
.	NN	O	O

Elf-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
lymphoid-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
belongs	NN	O	O
to	NN	O	O
the	NN	O	O
ETS	NN	O	B-protein
protein	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

It	NN	O	O
can	NN	O	O
bind	NN	O	O
to	NN	O	O
DNA	NN	O	O
target	NN	O	O
sequences	NN	O	O
within	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Elf-1	NN	O	B-protein
is	NN	O	O
constitutively	NN	O	O
localized	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
which	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
86-265	NN	O	I-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
Gal4-Elf-1	NN	O	B-protein
fusion	NN	O	I-protein
proteins	NN	O	I-protein
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
N-terminal	NN	O	O
86	NN	O	O
amino	NN	O	O
acids	NN	O	O
of	NN	O	O
Elf-1	NN	O	B-protein
contain	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
attenuated	NN	O	O
by	NN	O	O
an	NN	O	O
internal	NN	O	B-protein
repression	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
Elf-1	NN	O	B-protein
interacts	NN	O	O
specifically	NN	O	O
with	NN	O	O
the	NN	O	O
E74	NN	O	B-DNA
target	NN	O	I-DNA
sequence	NN	O	I-DNA
and	NN	O	O
can	NN	O	O
stimulate	NN	O	O
transcription	NN	O	O
driven	NN	O	O
by	NN	O	O
the	NN	O	O
E74	NN	O	B-DNA
site	NN	O	I-DNA
independent	NN	O	O
of	NN	O	O
mitogenic	NN	O	O
signaling	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Elf-1	NN	O	B-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
stimulate	NN	O	O
gene	NN	O	O
transcription	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
and	NN	O	O
activity	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
transduced	NN	O	O
with	NN	O	O
HTLV-I	NN	O	B-protein
Tax	NN	O	I-protein
mutants	NN	O	I-protein
with	NN	O	O
different	NN	O	O
trans-activating	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

Tax1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
trans-activator	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
,	NN	O	O
induces	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
cellular	NN	O	O
genes	NN	O	O
through	NN	O	O
interaction	NN	O	O
with	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
distinct	NN	O	O
cellular	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
;	NN	O	O
CREB/ATF	NN	O	B-protein
,	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
SRF	NN	O	B-protein
.	NN	O	O

This	NN	O	O
Tax1	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
critical	NN	O	O
event	NN	O	O
in	NN	O	O
T-cell	NN	O	O
transformation	NN	O	O
by	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
each	NN	O	O
Tax1	NN	O	B-protein
-inducible	NN	O	O
transcriptional	NN	O	O
pathway	NN	O	O
in	NN	O	O
T-cell	NN	O	O
transformation	NN	O	O
,	NN	O	O
we	NN	O	O
introduced	NN	O	O
Tax1	NN	O	B-protein
mutants	NN	O	I-protein
with	NN	O	O
different	NN	O	O
trans-activating	NN	O	O
phenotypes	NN	O	O
into	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
PBL	NN	O	B-cell_type
)	NN	O	O
by	NN	O	O
retroviral	NN	O	O
vectors	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
these	NN	O	O
PBLs	NN	O	B-cell_type
revealed	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
pathway	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
promote	NN	O	O
the	NN	O	O
growth	NN	O	O
response	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
for	NN	O	O
the	NN	O	O
clonal	NN	O	O
expansion	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
characteristic	NN	O	O
result	NN	O	O
of	NN	O	O
HTLV-I	NN	O	O
infection	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
CREB/ATF	NN	O	O
and	NN	O	O
SRF	NN	O	O
pathways	NN	O	O
is	NN	O	O
also	NN	O	O
required	NN	O	O
.	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	O
virus	NN	O	O
binding	NN	O	O
to	NN	O	O
CD21	NN	O	B-protein
activates	NN	O	O
the	NN	O	O
initial	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
via	NN	O	O
NF-kappaB	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
oncogenic	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
,	NN	O	O
binds	NN	O	O
to	NN	O	O
and	NN	O	O
infects	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
via	NN	O	O
CD21	NN	O	B-protein
,	NN	O	O
the	NN	O	O
CR2	NN	O	B-protein
complement	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
enable	NN	O	O
EBV	NN	O	O
to	NN	O	O
infect	NN	O	O
nonactivated	NN	O	O
,	NN	O	O
noncycling	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
provide	NN	O	O
compelling	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
sequence	NN	O	O
of	NN	O	O
events	NN	O	O
in	NN	O	O
which	NN	O	O
EBV	NN	O	O
binding	NN	O	O
to	NN	O	O
CD21	NN	O	B-protein
on	NN	O	O
purified	NN	O	O
resting	NN	O	B-cell_type
human	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
rapidly	NN	O	O
activates	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
,	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
binds	NN	O	O
to	NN	O	O
and	NN	O	O
mediates	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
Wp	NN	O	O
,	NN	O	O
the	NN	O	O
initial	NN	O	O
viral	NN	O	B-DNA
latent	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
EBV	NN	O	O
binding	NN	O	O
to	NN	O	O
its	NN	O	O
cellular	NN	O	O
receptor	NN	O	O
on	NN	O	O
resting	NN	O	O
B	NN	O	O
cells	NN	O	O
triggers	NN	O	O
an	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
which	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Synthetic	NN	O	O
glucocorticoids	NN	O	O
that	NN	O	O
dissociate	NN	O	O
transactivation	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
transrepression	NN	O	O
exhibit	NN	O	O
antiinflammatory	NN	O	O
activity	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Some	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
potent	NN	O	O
antiinflammatory	NN	O	O
and	NN	O	O
immunosuppressive	NN	O	O
agents	NN	O	O
are	NN	O	O
synthetic	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
major	NN	O	O
side	NN	O	O
effects	NN	O	O
severely	NN	O	O
limit	NN	O	O
their	NN	O	O
therapeutic	NN	O	O
use	NN	O	O
.	NN	O	O

The	NN	O	O
development	NN	O	O
of	NN	O	O
improved	NN	O	O
glucocorticoid-based	NN	O	O
drugs	NN	O	O
will	NN	O	O
require	NN	O	O
the	NN	O	O
separation	NN	O	O
of	NN	O	O
beneficial	NN	O	O
from	NN	O	O
deleterious	NN	O	O
effects	NN	O	O
.	NN	O	O

One	NN	O	O
possibility	NN	O	O
toward	NN	O	O
this	NN	O	O
goal	NN	O	O
is	NN	O	O
to	NN	O	O
try	NN	O	O
to	NN	O	O
dissociate	NN	O	O
two	NN	O	O
main	NN	O	O
activities	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
i.e.	NN	O	O
transactivation	NN	O	O
and	NN	O	O
transrepression	NN	O	O
.	NN	O	O

Screening	NN	O	O
of	NN	O	O
a	NN	O	O
library	NN	O	O
of	NN	O	O
compounds	NN	O	O
using	NN	O	O
transactivation	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
transrepression	NN	O	O
models	NN	O	O
in	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
cells	NN	O	O
identified	NN	O	O
dissociated	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
which	NN	O	O
exert	NN	O	O
strong	NN	O	O
AP-1	NN	O	B-protein
inhibition	NN	O	O
but	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
transactivation	NN	O	O
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
despite	NN	O	O
high	NN	O	O
ligand	NN	O	O
binding	NN	O	O
affinity	NN	O	O
,	NN	O	O
the	NN	O	O
prototypic	NN	O	O
dissociated	NN	O	O
compound	NN	O	O
,	NN	O	O
RU24858	NN	O	O
,	NN	O	O
acted	NN	O	O
as	NN	O	O
a	NN	O	O
weak	NN	O	O
agonist	NN	O	O
and	NN	O	O
did	NN	O	O
not	NN	O	O
efficiently	NN	O	O
antagonize	NN	O	O
dexamethasone-induced	NN	O	O
transcription	NN	O	O
in	NN	O	O
transfected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Similar	NN	O	O
results	NN	O	O
were	NN	O	O
obtained	NN	O	O
in	NN	O	O
hepatic	NN	O	B-cell_line
HTC	NN	O	I-cell_line
cells	NN	O	I-cell_line
for	NN	O	O
the	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
tyrosine	NN	O	I-DNA
amino	NN	O	I-DNA
transferase	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
TAT	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
enzymes	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
glucocorticoid-dependent	NN	O	O
stimulation	NN	O	O
of	NN	O	O
neoglucogenesis	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
whether	NN	O	O
dissociated	NN	O	O
glucocorticoids	NN	O	O
retained	NN	O	O
the	NN	O	O
antiinflammatory	NN	O	O
and	NN	O	O
immunosuppressive	NN	O	O
potential	NN	O	O
of	NN	O	O
classic	NN	O	O
glucocorticoids	NN	O	O
,	NN	O	O
several	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
models	NN	O	O
were	NN	O	O
used	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
secretion	NN	O	O
of	NN	O	O
the	NN	O	O
proinflammatory	NN	O	B-protein
lymphokine	NN	O	I-protein
interleukin-1beta	NN	O	I-protein
was	NN	O	O
severely	NN	O	O
inhibited	NN	O	O
by	NN	O	O
dissociated	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
THP	NN	O	I-cell_line
1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
in	NN	O	O
two	NN	O	O
in	NN	O	O
vivo	NN	O	O
models	NN	O	O
,	NN	O	O
these	NN	O	O
compounds	NN	O	O
exerted	NN	O	O
an	NN	O	O
antiinflammatory	NN	O	O
and	NN	O	O
immunosuppressive	NN	O	O
activity	NN	O	O
as	NN	O	O
potent	NN	O	O
as	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
classic	NN	O	O
glucocorticoid	NN	O	O
prednisolone	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
an	NN	O	O
improvement	NN	O	O
of	NN	O	O
antiinflammatory	NN	O	O
and	NN	O	O
immunosuppressive	NN	O	O
therapies	NN	O	O
and	NN	O	O
provide	NN	O	O
a	NN	O	O
novel	NN	O	O
concept	NN	O	O
for	NN	O	O
drug	NN	O	O
discovery	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
footprinting	NN	O	O
and	NN	O	O
mutational	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
CD19	NN	O	I-DNA
promoter	NN	O	I-DNA
reveal	NN	O	O
important	NN	O	O
roles	NN	O	O
for	NN	O	O
an	NN	O	O
SP1/Egr-1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
novel	NN	O	O
site	NN	O	O
termed	NN	O	O
the	NN	O	O
PyG	NN	O	B-DNA
box	NN	O	I-DNA
.	NN	O	O

CD19	NN	O	B-protein
expression	NN	O	O
begins	NN	O	O
at	NN	O	O
the	NN	O	O
pro-B	NN	O	O
cell	NN	O	O
stage	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

As	NN	O	O
such	NN	O	O
it	NN	O	O
serves	NN	O	O
as	NN	O	O
a	NN	O	O
good	NN	O	O
prototype	NN	O	O
for	NN	O	O
B	NN	O	B-DNA
cell-specific	NN	O	I-DNA
genes	NN	O	I-DNA
whose	NN	O	O
expression	NN	O	O
begins	NN	O	O
shortly	NN	O	O
after	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
controlling	NN	O	O
CD19	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
and	NN	O	O
functionally	NN	O	O
characterized	NN	O	O
the	NN	O	O
CD19	NN	O	B-DNA
promoter	NN	O	I-DNA
using	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprinting	NN	O	O
,	NN	O	O
gel	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
and	NN	O	O
transfection	NN	O	O
studies	NN	O	O
.	NN	O	O

Reporter	NN	O	O
constructs	NN	O	O
spanning	NN	O	O
portions	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
identified	NN	O	O
a	NN	O	O
region	NN	O	B-DNA
between	NN	O	I-DNA
-85	NN	O	I-DNA
and	NN	O	I-DNA
-200	NN	O	I-DNA
that	NN	O	O
produced	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
reporter	NN	O	O
gene	NN	O	O
activity	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
footprinting	NN	O	O
identified	NN	O	O
protected	NN	O	O
regions	NN	O	O
over	NN	O	O
the	NN	O	O
known	NN	O	O
high	NN	O	O
affinity	NN	O	O
B	NN	O	B-DNA
cell	NN	O	I-DNA
lineage-specific	NN	O	I-DNA
activator	NN	O	I-DNA
protein	NN	O	I-DNA
(	NN	O	I-DNA
BSAP	NN	O	I-DNA
)	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
low	NN	O	B-DNA
affinity	NN	O	I-DNA
BSAP	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
SP1/Egr-1	NN	O	B-DNA
site	NN	O	I-DNA
termed	NN	O	O
the	NN	O	O
CD19	NN	O	B-DNA
GC	NN	O	I-DNA
box	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
two	NN	O	O
novel	NN	O	O
sites	NN	O	O
named	NN	O	O
the	NN	O	O
AT	NN	O	B-DNA
box	NN	O	I-DNA
and	NN	O	O
PyG	NN	O	B-DNA
box	NN	O	I-DNA
.	NN	O	O

Phorbol	NN	O	O
ester	NN	O	O
treatment	NN	O	O
of	NN	O	O
a	NN	O	O
pre-B	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
up-regulated	NN	O	O
CD19	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
induced	NN	O	O
Egr-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
enhanced	NN	O	O
the	NN	O	O
footprint	NN	O	O
over	NN	O	O
the	NN	O	O
GC	NN	O	O
box	NN	O	O
.	NN	O	O

Gel	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
SP1	NN	O	O
and	NN	O	O
Egr-1	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
CD19	NN	O	B-DNA
GC	NN	O	I-DNA
box	NN	O	I-DNA
,	NN	O	O
while	NN	O	O
unknown	NN	O	O
nuclear	NN	O	O
proteins	NN	O	O
bound	NN	O	O
the	NN	O	O
PyG	NN	O	B-DNA
and	NN	O	I-DNA
AT	NN	O	I-DNA
boxes	NN	O	I-DNA
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
AT	NN	O	B-DNA
box	NN	O	I-DNA
or	NN	O	O
in	NN	O	O
the	NN	O	O
BSAP	NN	O	B-DNA
sites	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
affect	NN	O	O
CD19	NN	O	B-protein
reporter	NN	O	B-DNA
construct	NN	O	I-DNA
activity	NN	O	O
,	NN	O	O
while	NN	O	O
a	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
GC	NN	O	B-DNA
box	NN	O	I-DNA
reduced	NN	O	O
it	NN	O	O
modestly	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
PyG	NN	O	B-DNA
box	NN	O	I-DNA
mutation	NN	O	O
reduced	NN	O	O
it	NN	O	O
dramatically	NN	O	O
.	NN	O	O

BSAP	NN	O	B-protein
failed	NN	O	O
to	NN	O	O
trans-activate	NN	O	O
CD19	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
non-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
cis	NN	O	O
elements	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
PyG	NN	O	B-DNA
and	NN	O	I-DNA
GC	NN	O	I-DNA
boxes	NN	O	I-DNA
are	NN	O	O
also	NN	O	O
necessary	NN	O	O
for	NN	O	O
high	NN	O	O
level	NN	O	O
CD19	NN	O	B-DNA
promoter	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

EBF	NN	O	B-protein
and	NN	O	O
E47	NN	O	B-protein
collaborate	NN	O	O
to	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
surrogate	NN	O	I-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Early	NN	O	B-protein
B	NN	O	I-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EBF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
E47	NN	O	B-protein
participate	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
early	NN	O	O
B	NN	O	O
lymphocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

With	NN	O	O
the	NN	O	O
aim	NN	O	O
of	NN	O	O
identifying	NN	O	O
genetic	NN	O	O
targets	NN	O	O
for	NN	O	O
these	NN	O	O
transcription	NN	O	O
factors	NN	O	O
,	NN	O	O
we	NN	O	O
stably	NN	O	O
transfected	NN	O	B-DNA
cDNAs	NN	O	I-DNA
encoding	NN	O	O
EBF	NN	O	B-protein
or	NN	O	O
a	NN	O	O
covalent	NN	O	O
homodimer	NN	O	O
of	NN	O	O
E47	NN	O	B-protein
,	NN	O	O
individually	NN	O	O
or	NN	O	O
together	NN	O	O
,	NN	O	O
into	NN	O	O
immature	NN	O	B-cell_line
hematopoietic	NN	O	I-cell_line
Ba/F3	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
lack	NN	O	O
both	NN	O	O
factors	NN	O	O
.	NN	O	O

In	NN	O	O
combination	NN	O	O
,	NN	O	O
EBF	NN	O	B-protein
and	NN	O	O
E47	NN	O	B-protein
induce	NN	O	O
efficient	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
surrogate	NN	O	I-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
lambda5	NN	O	B-DNA
and	NN	O	O
VpreB	NN	O	B-DNA
,	NN	O	O
whereas	NN	O	O
other	NN	O	O
pre-B	NN	O	B-DNA
cell-specific	NN	O	I-DNA
genes	NN	O	I-DNA
remain	NN	O	O
silent	NN	O	O
.	NN	O	O

Multiple	NN	O	O
functionally	NN	O	O
important	NN	O	O
EBF	NN	O	B-DNA
and	NN	O	I-DNA
E47	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
lambda5	NN	O	B-DNA
promoter/enhancer	NN	O	I-DNA
region	NN	O	I-DNA
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
lambda5	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
direct	NN	O	O
genetic	NN	O	O
target	NN	O	O
for	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
EBF	NN	O	B-protein
and	NN	O	O
E47	NN	O	B-protein
synergize	NN	O	O
to	NN	O	O
activate	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
genes	NN	O	O
that	NN	O	O
define	NN	O	O
an	NN	O	O
early	NN	O	O
stage	NN	O	O
of	NN	O	O
the	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Cytokines	NN	O	B-protein
:	NN	O	O
shared	NN	O	O
receptors	NN	O	B-protein
,	NN	O	O
distinct	NN	O	O
functions	NN	O	O
.	NN	O	O

That	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
used	NN	O	O
by	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
IL-2	NN	O	B-protein
and	NN	O	O
IL-15	NN	O	B-protein
are	NN	O	O
identical	NN	O	O
would	NN	O	O
suggest	NN	O	O
that	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
have	NN	O	O
redundant	NN	O	O
roles	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
development	NN	O	O
;	NN	O	O
instead	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
is	NN	O	O
the	NN	O	O
guardian	NN	O	O
of	NN	O	O
thymus-derived	NN	O	O
T-cell	NN	O	O
homeostasis	NN	O	O
,	NN	O	O
while	NN	O	O
interleukin-15	NN	O	B-protein
promotes	NN	O	O
extrathymic	NN	O	O
development	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
IL-4	NN	O	B-protein
receptor	NN	O	I-protein
alpha-chain	NN	O	I-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
JAK-1	NN	O	B-protein
and	NN	O	O
STAT6	NN	O	B-protein
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
common	NN	O	B-protein
gamma-chain	NN	O	I-protein
(	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
functional	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
,	NN	O	O
yet	NN	O	O
cells	NN	O	O
lacking	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
are	NN	O	O
able	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

This	NN	O	O
has	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
suggestion	NN	O	O
that	NN	O	O
a	NN	O	O
surrogate	NN	O	B-protein
gamma'-chain	NN	O	I-protein
,	NN	O	O
which	NN	O	O
can	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
to	NN	O	O
mediate	NN	O	O
signaling	NN	O	O
,	NN	O	O
is	NN	O	O
expressed	NN	O	O
on	NN	O	O
cells	NN	O	O
lacking	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

An	NN	O	O
alternative	NN	O	O
possibility	NN	O	O
is	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
transduce	NN	O	O
signals	NN	O	O
by	NN	O	O
homodimerization	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
this	NN	O	O
latter	NN	O	O
possibility	NN	O	O
,	NN	O	O
a	NN	O	O
chimeric	NN	O	B-protein
receptor	NN	O	I-protein
containing	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
c-kit	NN	O	B-protein
(	NN	O	O
the	NN	O	O
stem	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
SCF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
and	NN	O	I-protein
transmembrane	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
was	NN	O	O
generated	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
receptor	NN	O	I-protein
kit/IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
with	NN	O	O
SCF	NN	O	B-protein
induces	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
alpha-associated	NN	O	I-protein
kinase	NN	O	I-protein
JAK-1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
STAT6	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
JAK-3	NN	O	B-protein
,	NN	O	O
which	NN	O	O
associates	NN	O	O
with	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
is	NN	O	O
not	NN	O	O
induced	NN	O	O
by	NN	O	O
SCF	NN	O	B-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

SCF	NN	O	B-protein
-mediated	NN	O	O
ligation	NN	O	O
of	NN	O	O
kit/IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
elicit	NN	O	O
IL-4	NN	O	B-protein
-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
including	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
CD23	NN	O	B-protein
and	NN	O	O
synthesis	NN	O	O
of	NN	O	O
germ-line	NN	O	B-RNA
epsilon	NN	O	I-RNA
transcripts	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
CTLL2	NN	O	I-cell_line
,	NN	O	O
ligation	NN	O	O
of	NN	O	O
kit/IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
induces	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
in	NN	O	O
JAK-1-deficient	NN	O	B-cell_line
HeLa	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
STAT6	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
is	NN	O	O
completely	NN	O	O
abolished	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
IL-4R	NN	O	B-protein
alpha	NN	O	I-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
activate	NN	O	O
JAK-1	NN	O	B-protein
and	NN	O	O
STAT6	NN	O	B-protein
and	NN	O	O
to	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-4	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
thus	NN	O	O
identifying	NN	O	O
a	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
IL-4	NN	O	B-protein
signaling	NN	O	O
can	NN	O	O
proceed	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
JAK-3	NN	O	B-protein
and	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Histamine	NN	O	O
modulates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
through	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
production	NN	O	O
via	NN	O	O
the	NN	O	O
H2	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U937	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
histamine	NN	O	O
and	NN	O	O
its	NN	O	O
agonists	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	I-DNA
c-myc	NN	O	I-DNA
proto-oncogenes	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
translational	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
U937	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Histamine	NN	O	O
transiently	NN	O	O
increased	NN	O	O
cAMP	NN	O	O
and	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
through	NN	O	O
H2	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Dibutyryl	NN	O	O
cAMP	NN	O	O
also	NN	O	O
increased	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	B-protein
,	NN	O	O
and	NN	O	O
levels	NN	O	O
remained	NN	O	O
elevated	NN	O	O
even	NN	O	O
after	NN	O	O
12	NN	O	O
hr	NN	O	O
of	NN	O	O
treatment	NN	O	O
.	NN	O	O

Dose-dependence	NN	O	O
studies	NN	O	O
using	NN	O	O
histamine	NN	O	O
and	NN	O	O
dimaprit	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
EC50	NN	O	O
values	NN	O	O
for	NN	O	O
cAMP	NN	O	O
production	NN	O	O
and	NN	O	O
c-fos	NN	O	B-DNA
increase	NN	O	O
were	NN	O	O
similar	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
cAMP	NN	O	O
might	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
c-fos	NN	O	B-DNA
induction	NN	O	O
via	NN	O	O
H2	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
studies	NN	O	O
carried	NN	O	O
out	NN	O	O
using	NN	O	O
H7	NN	O	B-protein
,	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A/protein	NN	O	I-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
inhibitor	NN	O	I-protein
,	NN	O	O
blocked	NN	O	O
c-fos	NN	O	B-DNA
induction	NN	O	O
,	NN	O	O
whereas	NN	O	O
no	NN	O	O
effect	NN	O	O
was	NN	O	O
observed	NN	O	O
with	NN	O	O
bisindolylmaleimide	NN	O	O
,	NN	O	O
a	NN	O	O
specific	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
inhibitor	NN	O	O
.	NN	O	O

No	NN	O	O
modification	NN	O	O
of	NN	O	O
c-myc	NN	O	B-DNA
expression	NN	O	O
could	NN	O	O
be	NN	O	O
detected	NN	O	O
on	NN	O	O
treatment	NN	O	O
with	NN	O	O
histamine	NN	O	O
or	NN	O	O
its	NN	O	O
analogues	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
dibutyryl	NN	O	O
cAMP	NN	O	O
induced	NN	O	O
a	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
this	NN	O	O
proto-oncogene	NN	O	B-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
dibutyryl	NN	O	O
cAMP	NN	O	O
inhibited	NN	O	O
cell	NN	O	O
growth	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
whereas	NN	O	O
histamine	NN	O	O
failed	NN	O	O
to	NN	O	O
affect	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Cells	NN	O	O
pretreated	NN	O	O
with	NN	O	O
dimaprit	NN	O	O
showed	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
cAMP	NN	O	O
response	NN	O	O
to	NN	O	O
subsequent	NN	O	O
addition	NN	O	O
of	NN	O	O
H2	NN	O	O
agonists	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
cAMP	NN	O	O
response	NN	O	O
to	NN	O	O
prostaglandin	NN	O	O
E2	NN	O	O
remained	NN	O	O
unaltered	NN	O	O
.	NN	O	O

This	NN	O	O
homologous	NN	O	O
mechanism	NN	O	O
of	NN	O	O
H2	NN	O	B-protein
receptor	NN	O	I-protein
desensitization	NN	O	O
was	NN	O	O
time	NN	O	O
dependent	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
histamine	NN	O	O
activates	NN	O	O
several	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
differentiation	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
cAMP	NN	O	O
and	NN	O	O
c-fos	NN	O	B-DNA
production	NN	O	O
,	NN	O	O
but	NN	O	O
fails	NN	O	O
to	NN	O	O
promote	NN	O	O
differentiation	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
apparently	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
rapid	NN	O	O
desensitization	NN	O	O
of	NN	O	O
H2	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
leukocytes	NN	O	B-cell_type
by	NN	O	O
binding	NN	O	O
of	NN	O	O
thrombin-stimulated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
tether	NN	O	O
and	NN	O	O
activate	NN	O	O
myeloid	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
potential	NN	O	O
relevance	NN	O	O
of	NN	O	O
this	NN	O	O
mechanism	NN	O	O
in	NN	O	O
acute	NN	O	O
myocardial	NN	O	O
infarction	NN	O	O
(	NN	O	O
AMI	NN	O	O
)	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
cytokine	NN	O	B-protein
induction	NN	O	O
by	NN	O	O
leukocyte-platelet	NN	O	O
adhesion	NN	O	O
and	NN	O	O
the	NN	O	O
occurrence	NN	O	O
of	NN	O	O
leukocyte-platelet	NN	O	O
conjugates	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
AMI	NN	O	O
.	NN	O	O

METHODS	NN	O	O
AND	NN	O	O
RESULTS	NN	O	O
:	NN	O	O
We	NN	O	O
obtained	NN	O	O
peripheral	NN	O	O
venous	NN	O	O
blood	NN	O	O
samples	NN	O	O
in	NN	O	O
20	NN	O	O
patients	NN	O	O
with	NN	O	O
AMI	NN	O	O
before	NN	O	O
and	NN	O	O
daily	NN	O	O
for	NN	O	O
5	NN	O	O
days	NN	O	O
after	NN	O	O
direct	NN	O	O
percutaneous	NN	O	O
transluminal	NN	O	O
coronary	NN	O	O
angioplasty	NN	O	O
(	NN	O	O
PTCA	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
20	NN	O	O
patients	NN	O	O
undergoing	NN	O	O
elective	NN	O	O
PTCA	NN	O	O
.	NN	O	O

Throughout	NN	O	O
the	NN	O	O
study	NN	O	O
period	NN	O	O
,	NN	O	O
CD41	NN	O	O
immunofluorescence	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
(	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
)	NN	O	O
revealed	NN	O	O
increased	NN	O	O
leukocyte-platelet	NN	O	O
adhesion	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
AMI	NN	O	O
compared	NN	O	O
with	NN	O	O
control	NN	O	O
patients	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SE	NN	O	O
of	NN	O	O
fluorescence	NN	O	O
[	NN	O	O
channels	NN	O	O
]	NN	O	O
before	NN	O	O
PTCA	NN	O	O
:	NN	O	O
77	NN	O	O
+/-	NN	O	O
16	NN	O	O
versus	NN	O	O
35	NN	O	O
+/-	NN	O	O
9	NN	O	O
;	NN	O	O
P	NN	O	O
=	NN	O	O
.003	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
,	NN	O	O
thrombin-stimulated	NN	O	B-cell_type
fixed	NN	O	I-cell_type
platelets	NN	O	I-cell_type
bound	NN	O	O
to	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Within	NN	O	O
2	NN	O	O
hours	NN	O	O
,	NN	O	O
this	NN	O	O
resulted	NN	O	O
in	NN	O	O
increased	NN	O	O
mRNA	NN	O	O
for	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	I-protein
1	NN	O	I-protein
beta	NN	O	I-protein
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
and	NN	O	O
monocyte	NN	O	B-protein
chemoattractant	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MCP	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
in	NN	O	O
unfractionated	NN	O	O
leukocytes	NN	O	B-cell_type
.	NN	O	O

After	NN	O	O
4	NN	O	O
hours	NN	O	O
,	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
IL-8	NN	O	B-protein
concentration	NN	O	O
of	NN	O	O
the	NN	O	O
cell-free	NN	O	O
supernatant	NN	O	O
had	NN	O	O
increased	NN	O	O
by	NN	O	O
268	NN	O	O
+/-	NN	O	O
36	NN	O	O
%	NN	O	O
and	NN	O	O
210	NN	O	O
+/-	NN	O	O
7	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
and	NN	O	O
cellular	NN	O	O
MCP-1	NN	O	B-protein
content	NN	O	O
had	NN	O	O
increased	NN	O	O
by	NN	O	O
170	NN	O	O
+/-	NN	O	O
8	NN	O	O
%	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
to	NN	O	O
adherent	NN	O	O
monocytes	NN	O	B-cell_type
had	NN	O	O
a	NN	O	O
similar	NN	O	O
effect	NN	O	O
and	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
binding	NN	O	O
by	NN	O	O
anti-P	NN	O	B-protein
selectin	NN	O	I-protein
antibodies	NN	O	I-protein
reduced	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
on	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
In	NN	O	O
patients	NN	O	O
with	NN	O	O
AMI	NN	O	O
,	NN	O	O
leukocyte-platelet	NN	O	O
adhesion	NN	O	O
is	NN	O	O
increased	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
induces	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
and	NN	O	O
MCP-1	NN	O	B-protein
in	NN	O	O
leukocytes	NN	O	B-cell_type
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
leukocyte-platelet	NN	O	O
adhesion	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
in	NN	O	O
AMI	NN	O	O
.	NN	O	O

-DOCSTART-	O

Immediate	NN	O	O
early	NN	O	O
and	NN	O	O
early	NN	O	B-protein
lytic	NN	O	I-protein
cycle	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
frequent	NN	O	O
targets	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	O
virus-induced	NN	O	O
cytotoxic	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
.	NN	O	O

Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
human	NN	O	O
gamma-herpesvirus	NN	O	O
,	NN	O	O
can	NN	O	O
establish	NN	O	O
both	NN	O	O
nonproductive	NN	O	O
(	NN	O	O
latent	NN	O	O
)	NN	O	O
and	NN	O	O
productive	NN	O	O
(	NN	O	O
lytic	NN	O	O
)	NN	O	O
infections	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
CD8+	NN	O	O
cytotoxic	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
(	NN	O	O
CTL	NN	O	O
)	NN	O	O
response	NN	O	O
to	NN	O	O
latently	NN	O	B-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
well	NN	O	O
characterized	NN	O	O
,	NN	O	O
very	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
T	NN	O	O
cell	NN	O	O
controls	NN	O	O
over	NN	O	O
lytic	NN	O	O
infection	NN	O	O
;	NN	O	O
this	NN	O	O
imbalance	NN	O	O
in	NN	O	O
our	NN	O	O
understanding	NN	O	O
belies	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
virus-replicative	NN	O	O
lesions	NN	O	O
in	NN	O	O
several	NN	O	O
aspects	NN	O	O
of	NN	O	O
EBV	NN	O	O
disease	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
work	NN	O	O
shows	NN	O	O
that	NN	O	O
the	NN	O	O
primary	NN	O	O
CD8+	NN	O	O
CTL	NN	O	O
response	NN	O	O
to	NN	O	O
EBV	NN	O	O
in	NN	O	O
infectious	NN	O	O
mononucleosis	NN	O	O
patients	NN	O	O
contains	NN	O	O
multiple	NN	O	O
lytic	NN	O	O
antigen-specific	NN	O	O
reactivities	NN	O	O
at	NN	O	O
levels	NN	O	O
at	NN	O	O
least	NN	O	O
as	NN	O	O
high	NN	O	O
as	NN	O	O
those	NN	O	O
seen	NN	O	O
against	NN	O	O
latent	NN	O	B-protein
antigens	NN	O	I-protein
;	NN	O	O
similar	NN	O	O
reactivities	NN	O	O
are	NN	O	O
also	NN	O	O
detectable	NN	O	O
in	NN	O	O
CTL	NN	O	O
memory	NN	O	O
.	NN	O	O

Clonal	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
individual	NN	O	O
responses	NN	O	O
to	NN	O	O
the	NN	O	O
two	NN	O	O
immediate	NN	O	B-protein
early	NN	O	I-protein
proteins	NN	O	I-protein
BZLF1	NN	O	B-protein
and	NN	O	O
BRLF1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
to	NN	O	O
three	NN	O	O
(	NN	O	O
BMLF1	NN	O	B-protein
,	NN	O	O
BMRF1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
BALF2	NN	O	B-protein
)	NN	O	O
of	NN	O	O
the	NN	O	O
six	NN	O	O
early	NN	O	B-protein
proteins	NN	O	I-protein
tested	NN	O	O
.	NN	O	O

In	NN	O	O
several	NN	O	O
cases	NN	O	O
,	NN	O	O
the	NN	O	O
peptide	NN	O	O
epitope	NN	O	O
and	NN	O	O
HLA-restricting	NN	O	O
determinant	NN	O	O
recognized	NN	O	O
by	NN	O	O
these	NN	O	O
CTLs	NN	O	B-cell_type
has	NN	O	O
been	NN	O	O
defined	NN	O	O
,	NN	O	O
one	NN	O	O
unusual	NN	O	O
feature	NN	O	O
being	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
responses	NN	O	O
restricted	NN	O	O
through	NN	O	O
HLA-C	NN	O	B-DNA
alleles	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
work	NN	O	O
strongly	NN	O	O
suggests	NN	O	O
that	NN	O	O
EBV-replicative	NN	O	O
lesions	NN	O	O
are	NN	O	O
subject	NN	O	O
to	NN	O	O
direct	NN	O	O
CTL	NN	O	O
control	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
that	NN	O	O
immediate	NN	O	B-protein
early	NN	O	I-protein
and	NN	O	I-protein
early	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
frequently	NN	O	O
the	NN	O	O
immunodominant	NN	O	O
targets	NN	O	O
.	NN	O	O

This	NN	O	O
contrasts	NN	O	O
with	NN	O	O
findings	NN	O	O
in	NN	O	O
alpha-	NN	O	O
and	NN	O	O
beta-	NN	O	O
herpesvirus	NN	O	O
systems	NN	O	O
(	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
,	NN	O	O
cytomegalovirus	NN	O	O
)	NN	O	O
where	NN	O	O
viral	NN	O	O
interference	NN	O	O
with	NN	O	O
the	NN	O	O
antigen-processing	NN	O	O
pathway	NN	O	O
during	NN	O	O
lytic	NN	O	O
infection	NN	O	O
renders	NN	O	O
immediate	NN	O	B-protein
early	NN	O	I-protein
and	NN	O	I-protein
early	NN	O	I-protein
proteins	NN	O	I-protein
much	NN	O	O
less	NN	O	O
immunogenic	NN	O	O
.	NN	O	O

The	NN	O	O
unique	NN	O	O
capacity	NN	O	O
of	NN	O	O
gamma-herpesvirus	NN	O	O
to	NN	O	O
amplify	NN	O	O
the	NN	O	O
viral	NN	O	O
load	NN	O	O
in	NN	O	O
vivo	NN	O	O
through	NN	O	O
a	NN	O	O
latent	NN	O	O
growth-transforming	NN	O	O
infection	NN	O	O
may	NN	O	O
have	NN	O	O
rendered	NN	O	O
these	NN	O	O
agents	NN	O	O
less	NN	O	O
dependent	NN	O	O
upon	NN	O	O
viral	NN	O	O
replication	NN	O	O
as	NN	O	O
a	NN	O	O
means	NN	O	O
of	NN	O	O
successfully	NN	O	O
colonizing	NN	O	O
their	NN	O	O
hosts	NN	O	O
.	NN	O	O

-DOCSTART-	O

Triggering	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
antigens	NN	O	I-protein
differentially	NN	O	O
modulates	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
release	NN	O	O
by	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
at	NN	O	O
distinct	NN	O	O
stage	NN	O	O
of	NN	O	O
maturation	NN	O	O
.	NN	O	O

Triggering	NN	O	O
of	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
by	NN	O	O
the	NN	O	O
anti-HLA-DR	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	I-protein
mAb	NN	O	I-protein
)	NN	O	I-protein
L243	NN	O	I-protein
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
and	NN	O	O
differentially	NN	O	O
enhanced	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
by	NN	O	O
the	NN	O	O
non-Hodgkin	NN	O	B-cell_type
's	NN	O	I-cell_type
lymphoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
Ri-I	NN	O	B-cell_line
,	NN	O	O
Ci-I	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
Sc-I	NN	O	B-cell_line
,	NN	O	O
which	NN	O	O
are	NN	O	O
at	NN	O	O
a	NN	O	O
distinct	NN	O	O
stage	NN	O	O
of	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
more	NN	O	O
mature	NN	O	O
Burkitt	NN	O	B-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
Raji	NN	O	I-cell_line
;	NN	O	O
in	NN	O	O
contrast	NN	O	O
,	NN	O	O
it	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
by	NN	O	O
the	NN	O	O
pre-B	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
Nalm-6	NN	O	B-cell_line
and	NN	O	O
BV173	NN	O	B-cell_line
.	NN	O	O

TNF-alpha	NN	O	B-protein
release	NN	O	O
peaked	NN	O	O
at	NN	O	O
24	NN	O	O
h	NN	O	O
and	NN	O	O
decreased	NN	O	O
thereafter	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
was	NN	O	O
dose	NN	O	O
dependent	NN	O	O
and	NN	O	O
preceded	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
detectable	NN	O	O
after	NN	O	O
3	NN	O	O
h	NN	O	O
of	NN	O	O
stimulation	NN	O	O
with	NN	O	O
mAb	NN	O	B-protein
L243	NN	O	I-protein
.	NN	O	O

Secreted	NN	O	O
TNF-alpha	NN	O	B-protein
mediated	NN	O	O
the	NN	O	O
enhancement	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
and	NN	O	O
activator	NN	O	O
protein-1	NN	O	O
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
binding	NN	O	O
activity	NN	O	O
;	NN	O	O
in	NN	O	O
fact	NN	O	O
,	NN	O	O
the	NN	O	O
triggering	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
antigens	NN	O	I-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
antihuman	NN	O	B-protein
TNF-alpha-neutralizing	NN	O	I-protein
antibodies	NN	O	I-protein
did	NN	O	O
not	NN	O	O
upregulate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
released	NN	O	O
TNF-alpha	NN	O	B-protein
was	NN	O	O
not	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
homotypic	NN	O	O
aggregation	NN	O	O
of	NN	O	O
Ri-I	NN	O	B-cell_line
,	NN	O	O
Ci-I	NN	O	B-cell_line
,	NN	O	O
Sc-I	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
Raji	NN	O	B-cell_line
cells	NN	O	I-cell_line
induced	NN	O	O
by	NN	O	O
mAb	NN	O	B-protein
L243	NN	O	I-protein
,	NN	O	O
and	NN	O	O
it	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
investigated	NN	O	O
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
:	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
release	NN	O	O
TNF-alpha	NN	O	B-protein
after	NN	O	O
triggering	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
antigens	NN	O	I-protein
depends	NN	O	O
on	NN	O	O
their	NN	O	O
stage	NN	O	O
of	NN	O	O
differentiation	NN	O	O
;	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
levels	NN	O	O
of	NN	O	O
released	NN	O	O
TNF-alpha	NN	O	B-protein
seem	NN	O	O
to	NN	O	O
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
stage	NN	O	O
of	NN	O	O
B-cell	NN	O	O
maturation	NN	O	O
but	NN	O	O
do	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
amounts	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
HLA-DR	NN	O	I-protein
antigens	NN	O	I-protein
;	NN	O	O
(	NN	O	O
c	NN	O	O
)	NN	O	O
secreted	NN	O	O
TNF-alpha	NN	O	B-protein
regulates	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
by	NN	O	O
an	NN	O	O
autocrine	NN	O	O
loop	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
d	NN	O	O
)	NN	O	O
intracellular	NN	O	O
signals	NN	O	O
mediating	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
by	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
regulating	NN	O	O
homotypic	NN	O	O
aggregation	NN	O	O
and	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-3	NN	O	I-protein
is	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_line
Th1	NN	O	I-cell_line
and	NN	O	I-cell_line
Th2	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
controls	NN	O	O
Th2-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-5	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Interleukin-5	NN	O	B-protein
(	NN	O	O
IL-5	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
produced	NN	O	O
by	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
2	NN	O	I-cell_type
(	NN	O	I-cell_type
Th2	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
eosinophilia	NN	O	O
in	NN	O	O
asthma	NN	O	O
.	NN	O	O

Despite	NN	O	O
increasing	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
outcome	NN	O	O
of	NN	O	O
many	NN	O	O
diseases	NN	O	O
is	NN	O	O
determined	NN	O	O
by	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
subsets	NN	O	O
of	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
Th1	NN	O	B-cell_type
and	NN	O	O
Th2	NN	O	B-cell_type
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
Th1-	NN	O	O
and	NN	O	O
Th2-	NN	O	O
specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
established	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-3	NN	O	B-protein
in	NN	O	O
IL-5	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
in	NN	O	O
EL-4	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
express	NN	O	O
both	NN	O	O
Th1-	NN	O	B-protein
and	NN	O	I-protein
Th2-type	NN	O	I-protein
cytokines	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
reported	NN	O	O
here	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
GATA-3	NN	O	B-protein
is	NN	O	O
critical	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-5	NN	O	B-protein
gene	NN	O	O
in	NN	O	O
bona	NN	O	B-cell_type
fide	NN	O	I-cell_type
Th2	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Whereas	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
GATA-3	NN	O	B-protein
site	NN	O	O
abolished	NN	O	O
antigen-	NN	O	O
or	NN	O	O
cAMP-	NN	O	O
stimulated	NN	O	O
IL-5	NN	O	O
promoter	NN	O	O
activation	NN	O	O
in	NN	O	O
Th2	NN	O	O
cells	NN	O	O
,	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
in	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
in	NN	O	O
a	NN	O	O
non-lymphoid	NN	O	B-cell_line
,	NN	O	I-cell_line
non-IL-5-producing	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
activated	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

During	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
T	NN	O	O
cell	NN	O	O
receptor	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
,	NN	O	O
GATA-3	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
up-regulated	NN	O	O
in	NN	O	O
developing	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
was	NN	O	O
down-regulated	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
antigen-	NN	O	B-cell_type
or	NN	O	I-cell_type
cAMP-activated	NN	O	I-cell_type
Th2	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
but	NN	O	O
not	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
expressed	NN	O	O
the	NN	O	O
GATA-3	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
GATA-3	NN	O	B-protein
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
balance	NN	O	O
between	NN	O	O
Th1	NN	O	B-cell_type
and	NN	O	O
Th2	NN	O	B-cell_type
subsets	NN	O	I-cell_type
in	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
activity	NN	O	O
has	NN	O	O
therapeutic	NN	O	O
potential	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
asthma	NN	O	O
and	NN	O	O
other	NN	O	O
hypereosinophilic	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
shortened	NN	O	O
life	NN	O	O
span	NN	O	O
of	NN	O	O
EKLF-/-	NN	O	B-cell_type
adult	NN	O	I-cell_type
erythrocytes	NN	O	I-cell_type
,	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
deficiency	NN	O	O
of	NN	O	O
beta-globin	NN	O	B-protein
chains	NN	O	I-protein
,	NN	O	O
is	NN	O	O
ameliorated	NN	O	O
by	NN	O	O
human	NN	O	B-protein
gamma-globin	NN	O	I-protein
chains	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
homologous	NN	O	O
recombination	NN	O	O
,	NN	O	O
both	NN	O	O
EKLF	NN	O	B-DNA
alleles	NN	O	I-DNA
in	NN	O	O
murine	NN	O	B-cell_line
embryonic	NN	O	I-cell_line
stem	NN	O	I-cell_line
(	NN	O	I-cell_line
ES	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
inactivated	NN	O	O
.	NN	O	O

These	NN	O	O
EKLF-/-	NN	O	B-cell_line
ES	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
capable	NN	O	O
of	NN	O	O
undergoing	NN	O	O
in	NN	O	O
vitro	NN	O	O
differentiation	NN	O	O
to	NN	O	O
form	NN	O	O
definitive	NN	O	O
erythroid	NN	O	O
colonies	NN	O	O
that	NN	O	O
were	NN	O	O
similar	NN	O	O
in	NN	O	O
size	NN	O	O
and	NN	O	O
number	NN	O	O
to	NN	O	O
those	NN	O	O
formed	NN	O	O
by	NN	O	O
wild-type	NN	O	B-cell_type
ES	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
EKLF-/-	NN	O	B-cell_line
colonies	NN	O	I-cell_line
were	NN	O	O
poorly	NN	O	O
hemoglobinized	NN	O	O
and	NN	O	O
enucleated	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
in	NN	O	O
these	NN	O	O
colonies	NN	O	O
contained	NN	O	O
numerous	NN	O	O
Heinz	NN	O	O
bodies	NN	O	O
.	NN	O	O

Reverse	NN	O	O
transcriptase-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
analyses	NN	O	O
revealed	NN	O	O
that	NN	O	O
adult	NN	O	O
and	NN	O	O
embryonic	NN	O	B-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
were	NN	O	O
appropriately	NN	O	O
regulated	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
beta	NN	O	B-protein
h1-globin	NN	O	I-protein
,	NN	O	O
which	NN	O	O
continued	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
at	NN	O	O
a	NN	O	O
very	NN	O	O
low	NN	O	O
level	NN	O	O
.	NN	O	O

The	NN	O	O
ratio	NN	O	O
of	NN	O	O
adult	NN	O	B-RNA
beta-globin/alpha-globin	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
mutant	NN	O	B-cell_line
ES	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
1/15	NN	O	O
of	NN	O	O
that	NN	O	O
in	NN	O	O
wild-type	NN	O	B-cell_type
ES	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

When	NN	O	O
the	NN	O	O
EKLF-/-	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
injected	NN	O	O
into	NN	O	O
blastocysts	NN	O	O
,	NN	O	O
they	NN	O	O
did	NN	O	O
not	NN	O	O
contribute	NN	O	O
at	NN	O	O
a	NN	O	O
detectable	NN	O	O
level	NN	O	O
to	NN	O	O
the	NN	O	O
mature	NN	O	O
erythrocyte	NN	O	O
compartment	NN	O	O
of	NN	O	O
the	NN	O	O
chimeric	NN	O	O
animals	NN	O	O
,	NN	O	O
based	NN	O	O
on	NN	O	O
analysis	NN	O	O
of	NN	O	O
glucose	NN	O	B-protein
phosphate	NN	O	I-protein
isomerase-1	NN	O	I-protein
(	NN	O	I-protein
GPI-1	NN	O	I-protein
)	NN	O	I-protein
isozymes	NN	O	I-protein
and	NN	O	O
hemoglobins	NN	O	O
that	NN	O	O
distinguish	NN	O	O
ES	NN	O	B-cell_type
cell-derived	NN	O	I-cell_type
erythrocytes	NN	O	I-cell_type
from	NN	O	O
host	NN	O	O
blastocyst-derived	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
semiquantitative	NN	O	O
RT-PCR	NN	O	O
analysis	NN	O	O
of	NN	O	O
RNA	NN	O	O
from	NN	O	O
reticulocytes	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
same	NN	O	O
chimeric	NN	O	O
animals	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
ES	NN	O	B-cell_type
cell-derived	NN	O	I-cell_type
reticulocytes	NN	O	I-cell_type
were	NN	O	O
present	NN	O	O
at	NN	O	O
a	NN	O	O
level	NN	O	O
of	NN	O	O
6	NN	O	O
%	NN	O	O
to	NN	O	O
8	NN	O	O
%	NN	O	O
.	NN	O	O

This	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
EKLF-/-	NN	O	B-cell_line
erythrocytes	NN	O	I-cell_line
in	NN	O	O
adult	NN	O	O
animals	NN	O	O
must	NN	O	O
be	NN	O	O
short-lived	NN	O	O
,	NN	O	O
apparently	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
imbalance	NN	O	O
of	NN	O	O
beta-	NN	O	B-protein
versus	NN	O	I-protein
alpha-globin	NN	O	I-protein
chains	NN	O	I-protein
,	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
precipitation	NN	O	O
of	NN	O	O
excess	NN	O	O
alpha-globin	NN	O	B-protein
chains	NN	O	I-protein
to	NN	O	O
form	NN	O	O
Heinz	NN	O	O
bodies	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
the	NN	O	O
short	NN	O	O
life	NN	O	O
span	NN	O	O
was	NN	O	O
ameliorated	NN	O	O
by	NN	O	O
introduction	NN	O	O
into	NN	O	O
the	NN	O	O
EKLF-/-	NN	O	B-cell_line
ES	NN	O	I-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
LCR/gamma-globin	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
ES	NN	O	B-cell_type
cell-derived	NN	O	I-cell_type
reticulocytes	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
mature	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
blood	NN	O	O
of	NN	O	O
the	NN	O	O
chimeric	NN	O	O
animals	NN	O	O
.	NN	O	O

-DOCSTART-	O

RP1	NN	O	B-DNA
,	NN	O	O
a	NN	O	O
new	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
adenomatous	NN	O	B-DNA
polyposis	NN	O	I-DNA
coli-binding	NN	O	I-DNA
EB1-like	NN	O	I-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cross-linking	NN	O	O
of	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
and	NN	O	I-protein
CD28	NN	O	I-protein
molecules	NN	O	I-protein
on	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
represents	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
effective	NN	O	O
signals	NN	O	O
for	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
activation	NN	O	O
and	NN	O	O
triggering	NN	O	O
of	NN	O	O
their	NN	O	O
cytotoxic	NN	O	O
effector	NN	O	O
function	NN	O	O
.	NN	O	O

To	NN	O	O
identify	NN	O	O
genes	NN	O	O
that	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
after	NN	O	O
stimulation	NN	O	O
,	NN	O	O
mRNA	NN	O	O
from	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
that	NN	O	O
had	NN	O	O
been	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
simultaneous	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
trigger	NN	O	O
molecules	NN	O	O
was	NN	O	O
transcribed	NN	O	O
for	NN	O	O
a	NN	O	O
differential	NN	O	O
mRNA	NN	O	O
display	NN	O	O
analysis	NN	O	O
into	NN	O	O
cDNA	NN	O	O
and	NN	O	O
was	NN	O	O
compared	NN	O	O
with	NN	O	O
cDNA	NN	O	O
from	NN	O	O
CD28	NN	O	B-protein
-or	NN	O	O
CD3	NN	O	B-protein
-activated	NN	O	O
or	NN	O	O
resting	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Differential	NN	O	O
expression	NN	O	O
was	NN	O	O
confirmed	NN	O	O
subsequently	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
cDNA	NN	O	B-DNA
fragments	NN	O	I-DNA
expressed	NN	O	O
specifically	NN	O	O
in	NN	O	O
CD3	NN	O	B-protein
-and	NN	O	O
CD28	NN	O	B-protein
-activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
designated	NN	O	O
RP1	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
predictive	NN	O	B-DNA
protein-coding	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
RP1	NN	O	B-DNA
had	NN	O	O
a	NN	O	O
significant	NN	O	O
homology	NN	O	O
to	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
recently	NN	O	O
found	NN	O	O
adenomatous	NN	O	B-DNA
polyposis	NN	O	I-DNA
coli	NN	O	I-DNA
(	NN	O	I-DNA
APC	NN	O	I-DNA
)	NN	O	I-DNA
protein-binding	NN	O	I-DNA
EB1	NN	O	I-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
codes	NN	O	O
for	NN	O	O
yet	NN	O	O
unknown	NN	O	O
protein	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
.	NN	O	O

Bacterially	NN	O	O
expressed	NN	O	O
RP1	NN	O	B-protein
protein	NN	O	I-protein
revealed	NN	O	O
specific	NN	O	O
binding	NN	O	O
to	NN	O	O
wild-type	NN	O	B-protein
but	NN	O	O
not	NN	O	O
to	NN	O	O
mutated	NN	O	B-protein
APC	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
rapid	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
RP1	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
properly	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
gene	NN	O	O
might	NN	O	O
belong	NN	O	O
to	NN	O	O
the	NN	O	O
immediate/early	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
controls	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
cascade	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
.	NN	O	O

As	NN	O	O
the	NN	O	O
expression	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
RP1	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
a	NN	O	O
spectrum	NN	O	O
of	NN	O	O
tumor-derived	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
proliferative	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
,	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
EB1-like	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
may	NN	O	O
not	NN	O	O
only	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
tumorigenesis	NN	O	O
of	NN	O	O
colorectal	NN	O	O
cancers	NN	O	O
but	NN	O	O
may	NN	O	O
also	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
proliferative	NN	O	O
control	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Cyclosporin	NN	O	O
A	NN	O	O
interferes	NN	O	O
with	NN	O	O
the	NN	O	O
inducible	NN	O	O
degradation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibitors	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
the	NN	O	O
processing	NN	O	O
of	NN	O	O
p105/	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
controls	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
numerous	NN	O	O
cytokine	NN	O	B-DNA
promoters	NN	O	I-DNA
during	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
by	NN	O	O
the	NN	O	O
immunosuppressants	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
and	NN	O	O
FK506	NN	O	O
exerts	NN	O	O
a	NN	O	O
suppressive	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
these	NN	O	O
NF-kappa	NN	O	B-DNA
B-controlled	NN	O	I-DNA
cytokine	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
show	NN	O	O
for	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
human	NN	O	B-cell_type
and	NN	O	I-cell_type
mouse	NN	O	I-cell_type
primary	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
that	NN	O	O
this	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
impaired	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
Rel	NN	O	B-protein
proteins	NN	O	I-protein
c-Rel	NN	O	B-protein
,	NN	O	O
RelA/p65	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B1/p50	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
nuclear	NN	O	O
appearance	NN	O	O
of	NN	O	O
RelB	NN	O	B-protein
remains	NN	O	O
unaffected	NN	O	O
.	NN	O	O

CsA	NN	O	O
does	NN	O	O
not	NN	O	O
interfere	NN	O	O
with	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
Rel	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
but	NN	O	O
prevents	NN	O	O
the	NN	O	O
inducible	NN	O	O
degradation	NN	O	O
of	NN	O	O
cytosolic	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
inhibitors	NN	O	I-protein
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
upon	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

CsA	NN	O	O
neither	NN	O	O
inhibits	NN	O	O
the	NN	O	O
processing	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
precursor	NN	O	I-protein
p105	NN	O	B-protein
to	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
nor	NN	O	O
does	NN	O	O
it	NN	O	O
``	NN	O	O
stabilize	NN	O	O
''	NN	O	O
the	NN	O	O
C-terminal	NN	O	O
portion	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
gamma	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
degraded	NN	O	O
during	NN	O	O
p105	NN	O	B-protein
processing	NN	O	O
to	NN	O	O
mature	NN	O	B-protein
p50	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
CsA	NN	O	O
interferes	NN	O	O
with	NN	O	O
a	NN	O	O
specific	NN	O	O
event	NN	O	O
in	NN	O	O
the	NN	O	O
signal-induced	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
beta	NN	O	I-protein
,	NN	O	O
but	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
processing	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B1/p105	NN	O	I-protein
to	NN	O	O
p50	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
by	NN	O	O
beta-amyloid	NN	O	O
peptides	NN	O	O
and	NN	O	O
interferon-gamma	NN	O	B-protein
in	NN	O	O
murine	NN	O	O
microglia	NN	O	O
.	NN	O	O

An	NN	O	O
increasing	NN	O	O
body	NN	O	O
of	NN	O	O
evidence	NN	O	O
suggests	NN	O	O
that	NN	O	O
amyloid-beta	NN	O	O
(	NN	O	O
A	NN	O	O
beta	NN	O	O
)	NN	O	O
peptides	NN	O	O
and	NN	O	O
microglia	NN	O	B-cell_type
are	NN	O	O
crucially	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
Alzheimer	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
effort	NN	O	O
to	NN	O	O
further	NN	O	O
elucidate	NN	O	O
the	NN	O	O
biological	NN	O	O
effects	NN	O	O
of	NN	O	O
A	NN	O	O
beta	NN	O	O
towards	NN	O	O
microglia	NN	O	B-cell_type
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
A	NN	O	O
beta	NN	O	O
peptides	NN	O	O
to	NN	O	O
activate	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
N9	NN	O	B-cell_line
murine	NN	O	I-cell_line
microglial	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Co-stimulation	NN	O	O
of	NN	O	O
microglia	NN	O	B-cell_type
with	NN	O	O
suboptimal	NN	O	O
concentrations	NN	O	O
of	NN	O	O
A	NN	O	O
beta	NN	O	O
(	NN	O	O
25-35	NN	O	O
)	NN	O	O
and	NN	O	O
100	NN	O	O
U/ml	NN	O	O
IFN	NN	O	B-protein
gamma	NN	O	I-protein
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
a	NN	O	O
specific	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
in	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

This	NN	O	O
response	NN	O	O
required	NN	O	O
at	NN	O	O
least	NN	O	O
120	NN	O	O
min	NN	O	O
to	NN	O	O
be	NN	O	O
evident	NN	O	O
and	NN	O	O
supershift	NN	O	O
experiments	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
contains	NN	O	O
both	NN	O	O
RelA	NN	O	B-protein
and	NN	O	O
p50	NN	O	B-protein
.	NN	O	O

Accordingly	NN	O	O
,	NN	O	O
immunoblot	NN	O	O
experiments	NN	O	O
showed	NN	O	O
that	NN	O	O
amongst	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
/Rel	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
RelA	NN	O	B-protein
and	NN	O	O
p50	NN	O	B-protein
are	NN	O	O
mobilized	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
following	NN	O	O
microglial	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
with	NN	O	O
A	NN	O	O
beta	NN	O	O
(	NN	O	O
25-35	NN	O	O
)	NN	O	O
plus	NN	O	O
IFN	NN	O	B-protein
gamma	NN	O	I-protein
.	NN	O	O

Higher	NN	O	O
concentrations	NN	O	O
of	NN	O	O
A	NN	O	O
beta	NN	O	O
(	NN	O	O
25-35	NN	O	O
)	NN	O	O
were	NN	O	O
effective	NN	O	O
by	NN	O	O
themselves	NN	O	O
in	NN	O	O
inducing	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
both	NN	O	O
in	NN	O	O
the	NN	O	O
N9	NN	O	B-cell_line
microglial	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
rat	NN	O	B-cell_line
primary	NN	O	I-cell_line
microglia	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

For	NN	O	O
purposes	NN	O	O
of	NN	O	O
comparison	NN	O	O
,	NN	O	O
microglia	NN	O	B-cell_type
were	NN	O	O
also	NN	O	O
stimulated	NN	O	O
with	NN	O	O
bacterial	NN	O	O
LPS	NN	O	O
,	NN	O	O
a	NN	O	O
known	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inducer	NN	O	O
.	NN	O	O

As	NN	O	O
expected	NN	O	O
,	NN	O	O
LPS	NN	O	O
strongly	NN	O	O
induced	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
two	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA-binding	NN	O	O
activities	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
RelA/p50	NN	O	B-protein
.	NN	O	O

The	NN	O	O
LPS	NN	O	O
response	NN	O	O
was	NN	O	O
also	NN	O	O
more	NN	O	O
rapid	NN	O	O
,	NN	O	O
as	NN	O	O
it	NN	O	O
was	NN	O	O
already	NN	O	O
evident	NN	O	O
by	NN	O	O
40	NN	O	O
min	NN	O	O
and	NN	O	O
remained	NN	O	O
sustained	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
3	NN	O	O
h	NN	O	O
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
might	NN	O	O
constitute	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
kappa	NN	O	B-DNA
B-dependent	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
microglia	NN	O	B-cell_type
stimulated	NN	O	O
by	NN	O	O
A	NN	O	O
beta	NN	O	O
peptides	NN	O	O
and	NN	O	O
IFN	NN	O	B-protein
gamma	NN	O	I-protein
,	NN	O	O
or	NN	O	O
by	NN	O	O
LPS	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid-mediated	NN	O	O
repression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
human	NN	O	O
arteritis-SCID	NN	O	B-protein
chimeras	NN	O	I-protein
.	NN	O	O

Giant	NN	O	O
cell	NN	O	O
arteritis	NN	O	O
(	NN	O	O
GCA	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
vasculitic	NN	O	O
syndrome	NN	O	O
that	NN	O	O
preferentially	NN	O	O
affects	NN	O	O
medium	NN	O	O
and	NN	O	O
large-sized	NN	O	O
arteries	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	O
therapy	NN	O	O
resolves	NN	O	O
clinical	NN	O	O
symptoms	NN	O	O
within	NN	O	O
hours	NN	O	O
to	NN	O	O
days	NN	O	O
,	NN	O	O
but	NN	O	O
therapy	NN	O	O
has	NN	O	O
to	NN	O	O
be	NN	O	O
continued	NN	O	O
over	NN	O	O
several	NN	O	O
years	NN	O	O
to	NN	O	O
prevent	NN	O	O
disease	NN	O	O
relapses	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
whether	NN	O	O
and	NN	O	O
how	NN	O	O
glucocorticoids	NN	O	O
affect	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
infiltrate	NN	O	O
or	NN	O	O
why	NN	O	O
the	NN	O	O
disease	NN	O	O
persists	NN	O	O
subclinically	NN	O	O
despite	NN	O	O
chronic	NN	O	O
treatment	NN	O	O
.	NN	O	O

GCA	NN	O	O
is	NN	O	O
self-sustained	NN	O	O
in	NN	O	O
temporal	NN	O	O
arteries	NN	O	O
engrafted	NN	O	O
into	NN	O	O
SCID	NN	O	O
mice	NN	O	O
,	NN	O	O
providing	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
action	NN	O	O
and	NN	O	O
limitations	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
can	NN	O	O
be	NN	O	O
examined	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Administration	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
to	NN	O	O
temporal	NN	O	B-cell_line
artery-SCID	NN	O	I-cell_line
chimeras	NN	O	I-cell_line
for	NN	O	O
1	NN	O	O
wk	NN	O	O
induced	NN	O	O
a	NN	O	O
partial	NN	O	O
suppression	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
and	NN	O	O
macrophage	NN	O	O
function	NN	O	O
as	NN	O	O
indicated	NN	O	O
by	NN	O	O
the	NN	O	O
reduced	NN	O	O
tissue	NN	O	O
concentrations	NN	O	O
of	NN	O	O
IL-2	NN	O	B-RNA
,	NN	O	I-RNA
IL-1beta	NN	O	I-RNA
,	NN	O	I-RNA
and	NN	O	I-RNA
IL-6	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
diminished	NN	O	O
expression	NN	O	O
of	NN	O	O
inducible	NN	O	O
NO	NN	O	B-protein
synthase	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
synthesis	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
only	NN	O	O
slightly	NN	O	O
decreased	NN	O	O
,	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
TGF-beta1	NN	O	B-protein
was	NN	O	O
unaffected	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
correlated	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IkappaBalpha	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
blockade	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
xenotransplanted	NN	O	O
tissue	NN	O	O
.	NN	O	O

Dose-response	NN	O	O
experiments	NN	O	O
suggested	NN	O	O
that	NN	O	O
steroid	NN	O	O
doses	NN	O	O
currently	NN	O	O
used	NN	O	O
in	NN	O	O
clinical	NN	O	O
medicine	NN	O	O
are	NN	O	O
suboptimal	NN	O	O
in	NN	O	O
repressing	NN	O	O
NFkappaB	NN	O	B-protein
-mediated	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
lesions	NN	O	O
.	NN	O	O

Chronic	NN	O	O
steroid	NN	O	O
therapy	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
deplete	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
products	NN	O	I-protein
IL-2	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
tissue-infiltrating	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
was	NN	O	O
only	NN	O	O
partially	NN	O	O
affected	NN	O	O
.	NN	O	O

IL-1beta	NN	O	B-protein
transcription	NN	O	O
was	NN	O	O
abrogated	NN	O	O
;	NN	O	O
in	NN	O	O
contrast	NN	O	O
,	NN	O	O
TGF-beta1	NN	O	B-RNA
mRNA	NN	O	I-RNA
synthesis	NN	O	O
was	NN	O	O
steroid	NN	O	O
resistant	NN	O	O
.	NN	O	O

The	NN	O	O
persistence	NN	O	O
of	NN	O	O
TGF-beta1-transcribing	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
despite	NN	O	O
paralysis	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
function	NN	O	O
,	NN	O	O
may	NN	O	O
provide	NN	O	O
an	NN	O	O
explanation	NN	O	O
for	NN	O	O
the	NN	O	O
chronicity	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
identify	NN	O	O
a	NN	O	O
novel	NN	O	O
therapeutic	NN	O	O
target	NN	O	O
in	NN	O	O
this	NN	O	O
inflammatory	NN	O	O
vasculopathy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Defective	NN	O	O
survival	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
expressing	NN	O	O
a	NN	O	O
catalytically	NN	O	B-protein
inactive	NN	O	I-protein
form	NN	O	I-protein
of	NN	O	O
Ca2+/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
IV	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
generated	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
that	NN	O	O
express	NN	O	O
a	NN	O	O
catalytically	NN	O	B-protein
inactive	NN	O	I-protein
form	NN	O	I-protein
of	NN	O	O
Ca2+/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
IV	NN	O	I-protein
(	NN	O	O
CaMKIV	NN	O	B-protein
)	NN	O	O
specifically	NN	O	O
in	NN	O	O
thymic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
thymic	NN	O	O
cellularity	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
the	NN	O	O
remaining	NN	O	O
cells	NN	O	O
is	NN	O	O
normal	NN	O	O
based	NN	O	O
on	NN	O	O
evaluation	NN	O	O
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
and	NN	O	O
CD8	NN	O	B-protein
cell	NN	O	B-protein
surface	NN	O	I-protein
antigens	NN	O	I-protein
that	NN	O	O
are	NN	O	O
used	NN	O	O
to	NN	O	O
gauge	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

Isolated	NN	O	O
thymic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
also	NN	O	O
show	NN	O	O
a	NN	O	O
dramatically	NN	O	O
decreased	NN	O	O
survival	NN	O	O
rate	NN	O	O
when	NN	O	O
evaluated	NN	O	O
in	NN	O	O
culture	NN	O	O
under	NN	O	O
conditions	NN	O	O
that	NN	O	O
do	NN	O	O
not	NN	O	O
favor	NN	O	O
activation	NN	O	O
.	NN	O	O

When	NN	O	O
challenged	NN	O	O
with	NN	O	O
an	NN	O	O
activating	NN	O	O
stimulus	NN	O	O
such	NN	O	O
as	NN	O	O
alpha-CD3	NN	O	B-protein
or	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
plus	NN	O	O
ionophore	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
are	NN	O	O
severely	NN	O	O
compromised	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
produce	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Reduction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
is	NN	O	O
secondary	NN	O	O
to	NN	O	O
the	NN	O	O
inability	NN	O	O
to	NN	O	O
phosphorylate	NN	O	O
the	NN	O	O
cAMP	NN	O	B-protein
response	NN	O	I-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
CREB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
genes	NN	O	I-DNA
such	NN	O	O
as	NN	O	O
Fos	NN	O	B-DNA
B	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
required	NN	O	O
to	NN	O	O
transactivate	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Because	NN	O	O
transgene	NN	O	O
expression	NN	O	O
was	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
lck	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
this	NN	O	O
promoter	NN	O	O
is	NN	O	O
inactivated	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
exit	NN	O	O
the	NN	O	O
thymus	NN	O	O
,	NN	O	O
the	NN	O	O
mutant	NN	O	B-protein
hCaMKIV	NN	O	I-protein
is	NN	O	O
not	NN	O	O
present	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
present	NN	O	O
in	NN	O	O
the	NN	O	O
spleen	NN	O	O
can	NN	O	O
be	NN	O	O
activated	NN	O	O
normally	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
either	NN	O	O
stimulus	NN	O	O
mentioned	NN	O	O
above	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
inactive	NN	O	B-protein
CaMKIV	NN	O	I-protein
on	NN	O	O
activation	NN	O	O
are	NN	O	O
reversible	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
CaMKIV	NN	O	B-protein
may	NN	O	O
represent	NN	O	O
a	NN	O	O
physiologically	NN	O	O
relevant	NN	O	O
CREB	NN	O	B-protein
kinase	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
the	NN	O	O
enzyme	NN	O	O
is	NN	O	O
also	NN	O	O
required	NN	O	O
to	NN	O	O
ensure	NN	O	O
normal	NN	O	O
expansion	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
.	NN	O	O

Whereas	NN	O	O
the	NN	O	O
pathway	NN	O	O
responsible	NN	O	O
for	NN	O	O
this	NN	O	O
latter	NN	O	O
role	NN	O	O
is	NN	O	O
yet	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
unlikely	NN	O	O
to	NN	O	O
include	NN	O	O
CREB	NN	O	B-protein
phosphorylation	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
new	NN	O	O
mouse	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
SRG3	NN	O	B-DNA
,	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
SWI3	NN	O	B-DNA
of	NN	O	O
Saccharomyces	NN	O	O
cerevisiae	NN	O	O
,	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
a	NN	O	O
thymoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
isolated	NN	O	O
a	NN	O	O
new	NN	O	O
mouse	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
highly	NN	O	O
expressed	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
,	NN	O	O
testis	NN	O	O
,	NN	O	O
and	NN	O	O
brain	NN	O	O
.	NN	O	O

This	NN	O	O
gene	NN	O	O
,	NN	O	O
SRG3	NN	O	B-DNA
,	NN	O	O
showed	NN	O	O
a	NN	O	O
significant	NN	O	O
sequence	NN	O	O
homology	NN	O	O
to	NN	O	O
SWI3	NN	O	B-DNA
,	NN	O	O
a	NN	O	O
yeast	NN	O	B-DNA
transcriptional	NN	O	I-DNA
activator	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
its	NN	O	O
human	NN	O	B-DNA
homolog	NN	O	I-DNA
BAF155	NN	O	I-DNA
.	NN	O	O

SRG3	NN	O	B-DNA
encodes	NN	O	O
1	NN	O	O
,	NN	O	O
100	NN	O	O
amino	NN	O	O
acids	NN	O	O
and	NN	O	O
has	NN	O	O
33-47	NN	O	O
%	NN	O	O
identity	NN	O	O
with	NN	O	O
SWI3	NN	O	B-protein
protein	NN	O	I-protein
over	NN	O	O
three	NN	O	O
regions	NN	O	O
.	NN	O	O

The	NN	O	O
SRG3	NN	O	B-protein
protein	NN	O	I-protein
contains	NN	O	O
an	NN	O	O
acidic	NN	O	B-protein
NH2	NN	O	I-protein
terminus	NN	O	I-protein
,	NN	O	O
a	NN	O	O
myb-like	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
a	NN	O	O
leucine-zipper	NN	O	B-protein
motif	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
proline-	NN	O	B-protein
and	NN	O	I-protein
glutamine-rich	NN	O	I-protein
region	NN	O	I-protein
at	NN	O	O
its	NN	O	O
COOH	NN	O	B-protein
terminus	NN	O	I-protein
.	NN	O	O

Rabbit	NN	O	O
antiserum	NN	O	O
raised	NN	O	O
against	NN	O	O
a	NN	O	O
COOH-terminal	NN	O	O
polypeptide	NN	O	O
of	NN	O	O
the	NN	O	O
SRG3	NN	O	B-DNA
recognized	NN	O	O
a	NN	O	O
protein	NN	O	O
with	NN	O	O
an	NN	O	O
apparent	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
155	NN	O	O
kD	NN	O	O
.	NN	O	O

The	NN	O	O
serum	NN	O	O
also	NN	O	O
detected	NN	O	O
a	NN	O	O
170-kD	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
mouse	NN	O	O
homologue	NN	O	O
of	NN	O	O
human	NN	O	B-protein
BAF170	NN	O	I-protein
.	NN	O	O

Immunoprecipitation	NN	O	O
of	NN	O	O
cell	NN	O	O
extract	NN	O	O
with	NN	O	O
the	NN	O	O
antiserum	NN	O	O
against	NN	O	O
the	NN	O	O
mouse	NN	O	B-protein
SRG3	NN	O	I-protein
also	NN	O	O
brought	NN	O	O
down	NN	O	O
a	NN	O	O
195-kD	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
could	NN	O	O
be	NN	O	O
recognized	NN	O	O
by	NN	O	O
an	NN	O	O
antiserum	NN	O	O
raised	NN	O	O
against	NN	O	O
human	NN	O	B-protein
SWI2	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
SRG3	NN	O	B-protein
protein	NN	O	I-protein
associates	NN	O	O
with	NN	O	O
a	NN	O	O
mouse	NN	O	B-protein
SWI2	NN	O	I-protein
.	NN	O	O

The	NN	O	O
SRG3	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
about	NN	O	O
three	NN	O	O
times	NN	O	O
higher	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
than	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
anti-sense	NN	O	B-protein
RNA	NN	O	I-protein
to	NN	O	O
SRG3	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
a	NN	O	O
thymoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
S49.1	NN	O	B-cell_line
,	NN	O	O
reduced	NN	O	O
the	NN	O	O
expression	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
SRG3	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
decreased	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
cell	NN	O	O
death	NN	O	O
induced	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
SRG3	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
glucocorticoid-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
the	NN	O	O
thymoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
implicates	NN	O	O
that	NN	O	O
the	NN	O	O
SRG3	NN	O	B-DNA
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
regulatory	NN	O	O
role	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
in	NN	O	O
thymus	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
collagenase-1	NN	O	B-protein
in	NN	O	O
differentiated	NN	O	B-cell_line
monocyte-like	NN	O	I-cell_line
(	NN	O	I-cell_line
U937	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
an	NN	O	O
upstream	NN	O	B-DNA
C/EBP-beta	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
site	NN	O	O
and	NN	O	O
a	NN	O	O
distal	NN	O	B-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
regulate	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
collagenase-1	NN	O	B-protein
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
expression	NN	O	I-DNA
constructs	NN	O	I-DNA
containing	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
collagenase-1	NN	O	I-DNA
promoter	NN	O	I-DNA
were	NN	O	O
stably	NN	O	O
or	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
into	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
reporter	NN	O	O
activity	NN	O	O
was	NN	O	O
assessed	NN	O	O
at	NN	O	O
various	NN	O	O
times	NN	O	O
after	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
-mediated	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Rapid	NN	O	O
and	NN	O	O
strong	NN	O	O
induction	NN	O	O
of	NN	O	O
promoter	NN	O	O
activity	NN	O	O
was	NN	O	O
lost	NN	O	O
in	NN	O	O
constructs	NN	O	O
with	NN	O	O
a	NN	O	O
mutant	NN	O	B-DNA
AP-1	NN	O	I-DNA
element	NN	O	I-DNA
;	NN	O	O
however	NN	O	O
,	NN	O	O
at	NN	O	O
16-96	NN	O	O
h	NN	O	O
post-PMA	NN	O	O
,	NN	O	O
the	NN	O	O
mutant	NN	O	B-DNA
collagenase-1	NN	O	I-DNA
promoter	NN	O	I-DNA
displayed	NN	O	O
AP-1	NN	O	B-protein
independent	NN	O	O
PMA-mediated	NN	O	O
transactivation	NN	O	O
.	NN	O	O

The	NN	O	O
AP-1	NN	O	B-DNA
mutant	NN	O	I-DNA
constructs	NN	O	I-DNA
also	NN	O	O
showed	NN	O	O
delayed	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
in	NN	O	O
PMA-treated	NN	O	B-cell_line
fibroblasts	NN	O	I-cell_line
.	NN	O	O

Western	NN	O	O
and	NN	O	O
supershift	NN	O	O
analyses	NN	O	O
indicated	NN	O	O
that	NN	O	O
functional	NN	O	B-protein
Jun	NN	O	I-protein
and	NN	O	I-protein
Fos	NN	O	I-protein
proteins	NN	O	I-protein
were	NN	O	O
present	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
PMA-differentiated	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Promoter	NN	O	O
deletion	NN	O	O
constructs	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
distal	NN	O	B-DNA
promoter	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
regulating	NN	O	O
collagenase-1	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
Western	NN	O	O
,	NN	O	O
supershift	NN	O	O
,	NN	O	O
and	NN	O	O
promoter	NN	O	O
deletion	NN	O	O
analyses	NN	O	O
suggested	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
CCAAT/enhancer-binding	NN	O	B-DNA
protein-beta	NN	O	I-DNA
(	NN	O	I-DNA
C/EBP-beta	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
between	NN	O	O
-2010	NN	O	B-DNA
and	NN	O	I-DNA
-1954	NN	O	I-DNA
in	NN	O	O
regulating	NN	O	O
transcription	NN	O	O
of	NN	O	O
collagenase-1	NN	O	B-protein
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
distinct	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
,	NN	O	O
acting	NN	O	O
somewhat	NN	O	O
independently	NN	O	O
of	NN	O	O
each	NN	O	O
other	NN	O	O
,	NN	O	O
control	NN	O	O
expression	NN	O	O
of	NN	O	O
collagenase-1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
rapid	NN	O	O
PMA-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
collagenase-1	NN	O	B-protein
transcription	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
mechanism	NN	O	O
distinct	NN	O	O
from	NN	O	O
that	NN	O	O
regulating	NN	O	O
the	NN	O	O
sustained	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
proteinase	NN	O	B-protein
in	NN	O	O
activated	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Quantification	NN	O	O
of	NN	O	O
vitamin	NN	O	B-RNA
D	NN	O	I-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
competitive	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
:	NN	O	O
lack	NN	O	O
of	NN	O	O
correspondence	NN	O	O
with	NN	O	O
common	NN	O	B-DNA
allelic	NN	O	I-DNA
variants	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
recently	NN	O	O
claimed	NN	O	O
that	NN	O	O
polymorphism	NN	O	O
for	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
influences	NN	O	O
several	NN	O	O
aspects	NN	O	O
of	NN	O	O
calcium	NN	O	O
and	NN	O	O
bone	NN	O	O
metabolism	NN	O	O
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
the	NN	O	O
physiologic	NN	O	O
plausibility	NN	O	O
of	NN	O	O
these	NN	O	O
claims	NN	O	O
,	NN	O	O
we	NN	O	O
compared	NN	O	O
the	NN	O	O
abundance	NN	O	O
of	NN	O	O
the	NN	O	O
VDR	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
between	NN	O	O
different	NN	O	O
VDR	NN	O	B-protein
genotypes	NN	O	I-protein
using	NN	O	O
a	NN	O	O
quantitative	NN	O	O
reverse	NN	O	O
transcribed	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction-based	NN	O	O
method	NN	O	O
.	NN	O	O

The	NN	O	O
method	NN	O	O
is	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
coamplification	NN	O	O
of	NN	O	O
VDR	NN	O	B-DNA
cDNA	NN	O	I-DNA
and	NN	O	O
an	NN	O	O
internal	NN	O	O
standard	NN	O	O
consisting	NN	O	O
of	NN	O	O
known	NN	O	O
concentrations	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
VDR	NN	O	I-DNA
CDNA	NN	O	I-DNA
mutated	NN	O	O
at	NN	O	O
a	NN	O	O
BglII	NN	O	B-DNA
restriction	NN	O	I-DNA
site	NN	O	I-DNA
;	NN	O	O
the	NN	O	O
interassay	NN	O	O
coefficient	NN	O	O
of	NN	O	O
variation	NN	O	O
is	NN	O	O
11	NN	O	O
%	NN	O	O
.	NN	O	O

To	NN	O	O
validate	NN	O	O
the	NN	O	O
method	NN	O	O
,	NN	O	O
we	NN	O	O
made	NN	O	O
use	NN	O	O
of	NN	O	O
earlier	NN	O	O
receptor	NN	O	O
binding	NN	O	O
studies	NN	O	O
indicating	NN	O	O
that	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
activated	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
resting	NN	O	O
,	NN	O	O
lymphocytes	NN	O	B-cell_type
expressed	NN	O	O
the	NN	O	O
VDR	NN	O	B-protein
.	NN	O	O

The	NN	O	O
concentration	NN	O	O
of	NN	O	O
the	NN	O	O
VDR	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
g/g	NN	O	O
of	NN	O	O
total	NN	O	O
RNA	NN	O	O
in	NN	O	O
cell-sorted	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
in	NN	O	B-cell_type
vitro	NN	O	I-cell_type
activated	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
only	NN	O	O
10	NN	O	O
(	NN	O	O
-12	NN	O	O
)	NN	O	O
g/g	NN	O	O
of	NN	O	O
total	NN	O	O
mRNA	NN	O	O
in	NN	O	O
resting	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
establishing	NN	O	O
that	NN	O	O
the	NN	O	O
VDR	NN	O	B-RNA
mRNA	NN	O	I-RNA
determined	NN	O	O
by	NN	O	O
our	NN	O	O
method	NN	O	O
in	NN	O	O
PBMCs	NN	O	B-cell_type
is	NN	O	O
due	NN	O	O
to	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Following	NN	O	O
an	NN	O	O
initial	NN	O	O
genotype	NN	O	O
screening	NN	O	O
of	NN	O	O
85	NN	O	O
normal	NN	O	O
volunteers	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
or	NN	O	O
restriction	NN	O	B-DNA
fragment	NN	O	I-DNA
length	NN	O	I-DNA
polymorphism	NN	O	I-DNA
analysis	NN	O	O
,	NN	O	O
14	NN	O	O
individuals	NN	O	O
with	NN	O	O
the	NN	O	O
Bb	NN	O	O
genotype	NN	O	O
,	NN	O	O
12	NN	O	O
with	NN	O	O
the	NN	O	O
bb	NN	O	O
genotype	NN	O	O
,	NN	O	O
and	NN	O	O
12	NN	O	O
with	NN	O	O
the	NN	O	O
BB	NN	O	O
genotype	NN	O	O
were	NN	O	O
selected	NN	O	O
.	NN	O	O

The	NN	O	O
concentration	NN	O	O
of	NN	O	O
the	NN	O	O
VDR	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
corrected	NN	O	O
for	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
was	NN	O	O
similar	NN	O	O
among	NN	O	O
the	NN	O	O
three	NN	O	O
genotype	NN	O	O
groups	NN	O	O
,	NN	O	O
as	NN	O	O
were	NN	O	O
the	NN	O	O
other	NN	O	O
variables	NN	O	O
examined	NN	O	O
:	NN	O	O
serum	NN	O	B-protein
calcitriol	NN	O	I-protein
,	NN	O	O
serum	NN	O	B-protein
osteocalcin	NN	O	I-protein
,	NN	O	O
and	NN	O	O
vertebral	NN	O	O
and	NN	O	O
hip	NN	O	O
bone	NN	O	O
density	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
VDR	NN	O	B-protein
polymorphism	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
abundance	NN	O	O
of	NN	O	O
the	NN	O	O
VDR	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

-DOCSTART-	O

Oncogenic	NN	O	O
forms	NN	O	O
of	NN	O	O
NOTCH1	NN	O	B-protein
lacking	NN	O	O
either	NN	O	O
the	NN	O	O
primary	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
for	NN	O	O
RBP-Jkappa	NN	O	B-protein
or	NN	O	O
nuclear	NN	O	B-protein
localization	NN	O	I-protein
sequences	NN	O	I-protein
retain	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
associate	NN	O	O
with	NN	O	O
RBP-Jkappa	NN	O	B-protein
and	NN	O	O
activate	NN	O	O
transcription	NN	O	O
.	NN	O	O

Truncated	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
NOTCH1	NN	O	B-protein
transmembrane	NN	O	I-protein
receptor	NN	O	I-protein
engineered	NN	O	O
to	NN	O	O
resemble	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
NOTCH1	NN	O	B-protein
found	NN	O	O
in	NN	O	O
certain	NN	O	O
cases	NN	O	O
of	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia/lymphoma	NN	O	O
(	NN	O	O
T-ALL	NN	O	O
)	NN	O	O
efficiently	NN	O	O
induce	NN	O	O
T-ALL	NN	O	O
when	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
bone	NN	O	O
marrow	NN	O	O
of	NN	O	O
mice	NN	O	O
.	NN	O	O

Unlike	NN	O	O
full-sized	NN	O	O
NOTCH1	NN	O	B-protein
,	NN	O	O
two	NN	O	O
such	NN	O	O
truncated	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
either	NN	O	O
lacking	NN	O	O
a	NN	O	O
major	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
domain	NN	O	I-protein
(	NN	O	O
DeltaE	NN	O	B-protein
)	NN	O	O
or	NN	O	O
consisting	NN	O	O
only	NN	O	O
of	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
domain	NN	O	I-protein
(	NN	O	O
ICN	NN	O	B-protein
)	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
in	NN	O	O
cultured	NN	O	O
cells	NN	O	O
,	NN	O	O
presumably	NN	O	O
through	NN	O	O
RBP-Jkappa	NN	O	B-protein
response	NN	O	O
elements	NN	O	O
within	NN	O	O
DNA	NN	O	O
.	NN	O	O

Both	NN	O	O
truncated	NN	O	O
forms	NN	O	O
also	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
RBP-Jkappa	NN	O	I-protein
in	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
from	NN	O	O
human	NN	O	B-cell_line
and	NN	O	I-cell_line
murine	NN	O	I-cell_line
T-ALL	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Transcriptional	NN	O	O
activation	NN	O	O
required	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
weak	NN	O	O
RBP-Jkappa-binding	NN	O	B-protein
site	NN	O	I-protein
within	NN	O	O
the	NN	O	O
NOTCH1	NN	O	B-protein
ankyrin	NN	O	I-protein
repeat	NN	O	I-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
a	NN	O	O
second	NN	O	O
,	NN	O	O
stronger	NN	O	O
RBP-Jkappa-binding	NN	O	B-protein
site	NN	O	I-protein
,	NN	O	O
which	NN	O	O
lies	NN	O	O
within	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
domain	NN	O	I-protein
close	NN	O	O
to	NN	O	O
the	NN	O	O
transmembrane	NN	O	O
region	NN	O	O
and	NN	O	O
significantly	NN	O	O
augments	NN	O	O
association	NN	O	O
with	NN	O	O
RBP-Jkappa	NN	O	B-protein
,	NN	O	O
was	NN	O	O
not	NN	O	O
needed	NN	O	O
for	NN	O	O
oncogenesis	NN	O	O
or	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

While	NN	O	O
ICN	NN	O	B-protein
appeared	NN	O	O
primarily	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
DeltaE	NN	O	B-protein
localized	NN	O	O
to	NN	O	O
cytoplasmic	NN	O	O
and	NN	O	O
nuclear	NN	O	O
membranes	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
intranuclear	NN	O	O
localization	NN	O	O
is	NN	O	O
not	NN	O	O
essential	NN	O	O
for	NN	O	O
oncogenesis	NN	O	O
or	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
support	NN	O	O
of	NN	O	O
this	NN	O	O
interpretation	NN	O	O
,	NN	O	O
mutation	NN	O	O
of	NN	O	O
putative	NN	O	O
nuclear	NN	O	B-protein
localization	NN	O	I-protein
sequences	NN	O	I-protein
decreased	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
and	NN	O	O
increased	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
membrane-bound	NN	O	B-protein
DeltaE	NN	O	I-protein
.	NN	O	O

Transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
this	NN	O	O
mutant	NN	O	O
form	NN	O	O
of	NN	O	O
membrane-bound	NN	O	B-protein
DeltaE	NN	O	I-protein
was	NN	O	O
approximately	NN	O	O
equivalent	NN	O	O
to	NN	O	O
that	NN	O	O
produced	NN	O	O
by	NN	O	O
intranuclear	NN	O	B-protein
ICN	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
most	NN	O	O
consistent	NN	O	O
with	NN	O	O
NOTCH1	NN	O	B-protein
oncogenesis	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
being	NN	O	O
independent	NN	O	O
of	NN	O	O
association	NN	O	O
with	NN	O	O
RBP-Jkappa	NN	O	B-protein
at	NN	O	O
promoter	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Sp	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
preferentially	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
proximal	NN	O	I-DNA
repeat	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Human	NN	O	O
T	NN	O	O
cell	NN	O	O
lymphotropic	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
encodes	NN	O	O
the	NN	O	O
transactivator	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
Tax	NN	O	B-protein
,	NN	O	O
which	NN	O	O
facilitates	NN	O	O
viral	NN	O	O
transcription	NN	O	O
from	NN	O	O
three	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeated	NN	O	I-DNA
elements	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
U3	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
the	NN	O	O
basal	NN	O	B-protein
factors	NN	O	I-protein
interacting	NN	O	O
with	NN	O	O
the	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeat	NN	O	I-DNA
elements	NN	O	I-DNA
through	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
(	NN	O	O
EMS	NN	O	O
)	NN	O	O
analyses	NN	O	O
has	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
DNA-protein	NN	O	B-protein
complexes	NN	O	I-protein
common	NN	O	O
to	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeats	NN	O	I-DNA
(	NN	O	O
C1-C3	NN	O	B-DNA
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
three	NN	O	O
DNA-protein	NN	O	B-protein
complexes	NN	O	I-protein
specific	NN	O	O
to	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
proximal	NN	O	I-DNA
(	NN	O	I-DNA
pp	NN	O	I-DNA
)	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
U1	NN	O	B-DNA
(	NN	O	O
U1A/U1B	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
U2	NN	O	B-DNA
;	NN	O	O
1-4	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
have	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
individual	NN	O	O
repeats	NN	O	O
are	NN	O	O
not	NN	O	O
identical	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
with	NN	O	O
which	NN	O	O
they	NN	O	O
interact	NN	O	O
.	NN	O	O

EMS	NN	O	O
analyses	NN	O	O
utilizing	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
mutated	NN	O	B-DNA
pp	NN	O	I-DNA
repeat	NN	O	I-DNA
elements	NN	O	I-DNA
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
requirements	NN	O	O
for	NN	O	O
U1	NN	O	B-DNA
(	NN	O	O
U1A/U1B	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
U2	NN	O	B-DNA
formation	NN	O	O
are	NN	O	O
separable	NN	O	O
from	NN	O	O
those	NN	O	O
required	NN	O	O
for	NN	O	O
C1-C3	NN	O	B-DNA
formation	NN	O	O
.	NN	O	O

Competition	NN	O	O
EMS	NN	O	O
analyses	NN	O	O
utilizing	NN	O	O
Sp1	NN	O	O
and	NN	O	O
CREB	NN	O	O
binding	NN	O	O
site	NN	O	O
oligonucleotides	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Sp	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
are	NN	O	O
critical	NN	O	O
components	NN	O	O
of	NN	O	O
U1	NN	O	B-DNA
(	NN	O	O
U1A/U1B	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
U2	NN	O	B-DNA
and	NN	O	O
that	NN	O	O
ATF/CREB	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
are	NN	O	O
critical	NN	O	O
components	NN	O	O
of	NN	O	O
C1-C3	NN	O	B-DNA
.	NN	O	O

EMS	NN	O	O
supershift	NN	O	O
analyses	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
Sp1	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
U1A	NN	O	O
formation	NN	O	O
while	NN	O	O
Sp3	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
U1B	NN	O	O
and	NN	O	O
U2	NN	O	O
formation	NN	O	O
.	NN	O	O

EMS	NN	O	O
analyses	NN	O	O
performed	NN	O	O
with	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
Tax-expressing	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
HTLV-I-transformed	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
demonstrate	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
prevents	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
U1	NN	O	B-DNA
(	NN	O	O
U1A/U1B	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
U2	NN	O	B-DNA
DNA-protein	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
Tax	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
Sp	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
with	NN	O	O
the	NN	O	O
pp	NN	O	B-DNA
repeat	NN	O	I-DNA
.	NN	O	O

Based	NN	O	O
on	NN	O	O
these	NN	O	O
observations	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
possible	NN	O	O
that	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
Sp	NN	O	B-protein
and	NN	O	O
ATF/CREB	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
with	NN	O	O
the	NN	O	O
pp	NN	O	B-DNA
repeat	NN	O	I-DNA
during	NN	O	O
basal	NN	O	O
and	NN	O	O
Tax	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
the	NN	O	O
initial	NN	O	O
stages	NN	O	O
of	NN	O	O
virus	NN	O	O
infection	NN	O	O
or	NN	O	O
during	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
latent	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lineage-	NN	O	O
and	NN	O	O
stage-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
runt	NN	O	B-protein
box	NN	O	I-protein
polypeptides	NN	O	I-protein
in	NN	O	O
primitive	NN	O	O
and	NN	O	O
definitive	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

Translocations	NN	O	O
involving	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CBFA2	NN	O	I-DNA
locus	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
leukemia	NN	O	O
.	NN	O	O

This	NN	O	O
gene	NN	O	O
,	NN	O	O
originally	NN	O	O
named	NN	O	O
AML1	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
a	NN	O	O
human	NN	O	O
homologue	NN	O	O
of	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-DNA
gene	NN	O	I-DNA
runt	NN	O	I-DNA
that	NN	O	O
controls	NN	O	O
early	NN	O	O
events	NN	O	O
in	NN	O	O
fly	NN	O	O
embryogenesis	NN	O	O
.	NN	O	O

To	NN	O	O
clarify	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
mammalian	NN	O	B-protein
runt	NN	O	I-protein
products	NN	O	I-protein
in	NN	O	O
normal	NN	O	O
and	NN	O	O
leukemic	NN	O	O
hematopoiesis	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
their	NN	O	O
pattern	NN	O	O
of	NN	O	O
expression	NN	O	O
in	NN	O	O
mouse	NN	O	O
hematopoietic	NN	O	O
tissues	NN	O	O
in	NN	O	O
the	NN	O	O
adult	NN	O	O
and	NN	O	O
during	NN	O	O
ontogeny	NN	O	O
using	NN	O	O
an	NN	O	O
anti-runt	NN	O	O
box	NN	O	O
antiserum	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
adult	NN	O	O
bone	NN	O	O
marrow	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
expression	NN	O	O
of	NN	O	O
runt	NN	O	O
polypeptides	NN	O	O
in	NN	O	O
differentiating	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Within	NN	O	O
the	NN	O	O
erythroid	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
runt	NN	O	O
expression	NN	O	O
is	NN	O	O
biphasic	NN	O	O
,	NN	O	O
clearly	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
erythroblasts	NN	O	B-cell_type
of	NN	O	O
early	NN	O	O
blood	NN	O	O
islands	NN	O	O
and	NN	O	O
of	NN	O	O
the	NN	O	O
fetal	NN	O	O
liver	NN	O	O
,	NN	O	O
but	NN	O	O
absent	NN	O	O
in	NN	O	O
the	NN	O	O
adult	NN	O	O
.	NN	O	O

Biochemical	NN	O	O
analysis	NN	O	O
by	NN	O	O
Western	NN	O	O
blotting	NN	O	O
of	NN	O	O
fetal	NN	O	B-cell_type
and	NN	O	I-cell_type
adult	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
populations	NN	O	I-cell_type
shows	NN	O	O
several	NN	O	O
runt	NN	O	B-protein
isoforms	NN	O	I-protein
.	NN	O	O

At	NN	O	O
least	NN	O	O
one	NN	O	O
of	NN	O	O
them	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
myeloid	NN	O	O
specific	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription-3	NN	O	I-protein
(	NN	O	O
STAT3	NN	O	B-protein
)	NN	O	O
is	NN	O	O
constitutively	NN	O	O
activated	NN	O	O
in	NN	O	O
normal	NN	O	O
,	NN	O	O
self-renewing	NN	O	B-cell_type
B-1	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
only	NN	O	O
inducibly	NN	O	O
expressed	NN	O	O
in	NN	O	O
conventional	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Cytokine	NN	O	B-protein
and	NN	O	O
growth	NN	O	B-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
engagement	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
rapid	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
latent	NN	O	O
,	NN	O	O
cytosolic	NN	O	B-protein
signal	NN	O	I-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
which	NN	O	O
then	NN	O	O
translocate	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
where	NN	O	O
they	NN	O	O
regulate	NN	O	O
transcriptional	NN	O	O
events	NN	O	O
from	NN	O	O
specific	NN	O	B-DNA
promoter	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

STAT3	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
particular	NN	O	O
has	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
Abl	NN	O	B-protein
,	NN	O	O
Src	NN	O	B-protein
,	NN	O	O
and	NN	O	O
HTLV-1	NN	O	O
transformation	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

B-1	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
are	NN	O	O
self-renewing	NN	O	O
,	NN	O	O
CD5+	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
display	NN	O	O
a	NN	O	O
propensity	NN	O	O
for	NN	O	O
malignant	NN	O	O
transformation	NN	O	O
and	NN	O	O
are	NN	O	O
the	NN	O	O
normal	NN	O	O
counterpart	NN	O	O
to	NN	O	O
human	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemias	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
B-1	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
characterized	NN	O	O
by	NN	O	O
aberrant	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
,	NN	O	O
including	NN	O	O
hyperresponsiveness	NN	O	O
to	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
PKC	NN	O	O
agonists	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
B-1	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
constitutively	NN	O	O
express	NN	O	O
nuclear	NN	O	B-protein
activated	NN	O	I-protein
STAT3	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
not	NN	O	O
expressed	NN	O	O
by	NN	O	O
unmanipulated	NN	O	B-cell_type
conventional	NN	O	I-cell_type
(	NN	O	I-cell_type
B-2	NN	O	I-cell_type
)	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
STAT3	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
induced	NN	O	O
in	NN	O	O
B-2	NN	O	B-cell_type
cells	NN	O	I-cell_type
after	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
engagement	NN	O	O
in	NN	O	O
a	NN	O	O
delayed	NN	O	O
fashion	NN	O	O
(	NN	O	O
after	NN	O	O
3	NN	O	O
h	NN	O	O
)	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
both	NN	O	O
the	NN	O	O
serine/threonine	NN	O	O
protein	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
H-7	NN	O	O
and	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
drug	NN	O	O
rapamycin	NN	O	O
and	NN	O	O
requires	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
novel	NN	O	O
coupling	NN	O	O
between	NN	O	O
sIg	NN	O	B-protein
and	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
differs	NN	O	O
from	NN	O	O
the	NN	O	O
classical	NN	O	O
paradigm	NN	O	O
for	NN	O	O
STAT	NN	O	B-protein
induction	NN	O	O
by	NN	O	O
cytokine	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
inability	NN	O	O
of	NN	O	O
prolonged	NN	O	O
stimulation	NN	O	O
of	NN	O	O
conventional	NN	O	O
B-2	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
anti-Ig	NN	O	B-protein
,	NN	O	O
a	NN	O	O
treatment	NN	O	O
sufficient	NN	O	O
to	NN	O	O
induce	NN	O	O
CD5	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
sustained	NN	O	O
STAT3	NN	O	B-protein
activation	NN	O	O
suggests	NN	O	O
that	NN	O	O
STAT3	NN	O	B-protein
is	NN	O	O
a	NN	O	O
specific	NN	O	O
nuclear	NN	O	O
marker	NN	O	O
for	NN	O	O
B-1	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
STAT3	NN	O	B-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
B	NN	O	O
cell	NN	O	O
antigen-specific	NN	O	O
signaling	NN	O	O
responses	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
normal	NN	O	O
cell	NN	O	O
population	NN	O	O
exhibiting	NN	O	O
intrinsic	NN	O	O
proliferative	NN	O	O
behavior	NN	O	O
.	NN	O	O

-DOCSTART-	O

Significance	NN	O	O
of	NN	O	O
quantitative	NN	O	O
analysis	NN	O	O
of	NN	O	O
AML1/ETO	NN	O	B-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
t	NN	O	O
(	NN	O	O
8	NN	O	O
;	NN	O	O
21	NN	O	O
)	NN	O	O
acute	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Autologous	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
stem	NN	O	I-cell_type
cell	NN	O	I-cell_type
transplantation	NN	O	O
(	NN	O	O
PBSCT	NN	O	O
)	NN	O	O
is	NN	O	O
replacing	NN	O	O
autologous	NN	O	O
bone	NN	O	O
marrow	NN	O	O
transplantation	NN	O	O
(	NN	O	O
BMT	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
leukemia	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
potential	NN	O	O
advantages	NN	O	O
of	NN	O	O
autologous	NN	O	O
PBSCT	NN	O	O
is	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBSC	NN	O	B-cell_type
)	NN	O	O
are	NN	O	O
less	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
contaminated	NN	O	O
by	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
than	NN	O	O
bone	NN	O	O
marrow	NN	O	O
grafts	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
major	NN	O	O
problem	NN	O	O
still	NN	O	O
remains	NN	O	O
the	NN	O	O
high	NN	O	O
incidence	NN	O	O
of	NN	O	O
leukemic	NN	O	O
relapse	NN	O	O
following	NN	O	O
autologous	NN	O	O
PBSCT	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
reinfusion	NN	O	O
of	NN	O	O
PBSC	NN	O	B-cell_type
contaminated	NN	O	O
by	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
quantitative	NN	O	O
assay	NN	O	O
using	NN	O	O
competitive	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
polymerase	NN	O	I-protein
chain	NN	O	O
reaction	NN	O	O
that	NN	O	O
estimates	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
AML1/ETO	NN	O	B-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
t	NN	O	O
(	NN	O	O
8	NN	O	O
;	NN	O	O
21	NN	O	O
)	NN	O	O
acute	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
(	NN	O	O
AML	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
leukemic	NN	O	O
cell	NN	O	O
contamination	NN	O	O
in	NN	O	O
PBSC	NN	O	B-cell_type
harvests	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
monitor	NN	O	O
minimal	NN	O	O
residual	NN	O	O
disease	NN	O	O
(	NN	O	O
MRD	NN	O	O
)	NN	O	O
quantitatively	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
t	NN	O	O
(	NN	O	O
8	NN	O	O
;	NN	O	O
21	NN	O	O
)	NN	O	O
AML	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
although	NN	O	O
PBSC	NN	O	B-cell_type
harvests	NN	O	O
collected	NN	O	O
after	NN	O	O
consolidation	NN	O	O
chemotherapy	NN	O	O
are	NN	O	O
contaminated	NN	O	O
by	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
leukemic	NN	O	O
cell	NN	O	O
contamination	NN	O	O
decreases	NN	O	O
with	NN	O	O
repeated	NN	O	O
cycles	NN	O	O
of	NN	O	O
chemotherapy	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
MRD	NN	O	O
in	NN	O	O
PBSC	NN	O	B-cell_type
harvests	NN	O	O
is	NN	O	O
less	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
corresponding	NN	O	O
bone	NN	O	O
marrow	NN	O	O
obtained	NN	O	O
on	NN	O	O
the	NN	O	O
day	NN	O	O
of	NN	O	O
the	NN	O	O
PBSC	NN	O	B-cell_type
collection	NN	O	O
.	NN	O	O

There	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
no	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
AML1/ETO	NN	O	B-RNA
transcripts	NN	O	I-RNA
found	NN	O	O
in	NN	O	O
the	NN	O	O
infused	NN	O	O
PBSC	NN	O	B-cell_type
harvests	NN	O	O
and	NN	O	O
the	NN	O	O
incidence	NN	O	O
of	NN	O	O
leukemic	NN	O	O
relapse	NN	O	O
following	NN	O	O
autologous	NN	O	O
PBSCT	NN	O	O
in	NN	O	O
our	NN	O	O
study	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
substantial	NN	O	O
decrease	NN	O	O
of	NN	O	O
AML1/ETO	NN	O	B-RNA
transcripts	NN	O	I-RNA
was	NN	O	O
seen	NN	O	O
following	NN	O	O
autologous	NN	O	O
PBSCT	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
quantitative	NN	O	O
analysis	NN	O	O
of	NN	O	O
AML1/ETO	NN	O	B-RNA
transcripts	NN	O	I-RNA
may	NN	O	O
be	NN	O	O
clinically	NN	O	O
useful	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
t	NN	O	O
(	NN	O	O
8	NN	O	O
;	NN	O	O
21	NN	O	O
)	NN	O	O
AML	NN	O	O
.	NN	O	O

-DOCSTART-	O

Upregulation	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
and	NN	O	I-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
human	NN	O	B-protein
immunodeficiency	NN	O	I-protein
virus-1	NN	O	I-protein
Tat	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
regulatory	NN	O	B-protein
Tat	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type-1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
viral	NN	O	O
replication	NN	O	O
and	NN	O	O
also	NN	O	O
shows	NN	O	O
pleiotropic	NN	O	O
activities	NN	O	O
on	NN	O	O
various	NN	O	O
cell	NN	O	O
functions	NN	O	O
.	NN	O	O

To	NN	O	O
get	NN	O	O
further	NN	O	O
insights	NN	O	O
into	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
the	NN	O	O
biological	NN	O	O
activity	NN	O	O
of	NN	O	O
Tat	NN	O	B-protein
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
endogenous	NN	O	O
and	NN	O	O
exogenous	NN	O	O
Tat	NN	O	B-protein
protein	NN	O	I-protein
on	NN	O	O
c-fos	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_line
lymphoblastoid	NN	O	I-cell_line
(	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
)	NN	O	I-cell_line
and	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
(	NN	O	I-cell_line
U937	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

Transient	NN	O	O
cotransfection	NN	O	O
of	NN	O	O
tat	NN	O	B-DNA
cDNA	NN	O	I-DNA
in	NN	O	O
sense	NN	O	O
orientation	NN	O	O
(	NN	O	O
tat/S	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
together	NN	O	O
with	NN	O	O
a	NN	O	O
plasmid	NN	O	B-DNA
containing	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
FC3	NN	O	B-DNA
,	NN	O	O
from-	NN	O	O
711	NN	O	B-DNA
to	NN	O	I-DNA
+42	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
front	NN	O	O
of	NN	O	O
the	NN	O	O
bacterial	NN	O	B-DNA
chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
significantly	NN	O	O
enhanced	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
activated	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
15	NN	O	O
%	NN	O	O
fetal	NN	O	O
calf	NN	O	O
serum	NN	O	O
(	NN	O	O
FCS	NN	O	O
)	NN	O	O
or	NN	O	O
5	NN	O	O
micrograms/mL	NN	O	O
phytohemagglutinin	NN	O	B-protein
plus	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
mol/L	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
activated	NN	O	O
by	NN	O	O
15	NN	O	O
%	NN	O	O
FCS	NN	O	O
or	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
mol/L	NN	O	O
PMA	NN	O	O
.	NN	O	O

This	NN	O	O
effect	NN	O	O
was	NN	O	O
specifically	NN	O	O
due	NN	O	O
to	NN	O	O
Tat	NN	O	B-protein
,	NN	O	O
since	NN	O	O
Jurkat	NN	O	B-cell_line
and	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
cotransfected	NN	O	O
either	NN	O	O
with	NN	O	O
tat	NN	O	B-DNA
cDNA	NN	O	I-DNA
in	NN	O	O
antisense	NN	O	O
orientation	NN	O	O
(	NN	O	O
tat/AS	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
tat	NN	O	B-DNA
carrying	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
aminoacid	NN	O	B-protein
cys22-gly22	NN	O	I-protein
(	NN	O	O
tat	NN	O	B-DNA
22/S	NN	O	I-DNA
)	NN	O	O
or	NN	O	O
with	NN	O	O
the	NN	O	O
backbone	NN	O	B-DNA
vector	NN	O	I-DNA
alone	NN	O	I-DNA
(	NN	O	O
pRPneo-SL3	NN	O	B-DNA
)	NN	O	O
did	NN	O	O
not	NN	O	O
show	NN	O	O
any	NN	O	O
significant	NN	O	O
difference	NN	O	O
in	NN	O	O
c-fos	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
FC3	NN	O	B-DNA
plasmid	NN	O	I-DNA
alone	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
deletion	NN	O	O
mutants	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
minimal	NN	O	B-DNA
DNA	NN	O	I-DNA
sequence	NN	O	I-DNA
required	NN	O	O
for	NN	O	O
Tat	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
located	NN	O	O
between	NN	O	O
nucleotides	NN	O	B-DNA
-404/-220	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
the	NN	O	O
serum	NN	O	B-DNA
responsive	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
SRE	NN	O	B-DNA
,	NN	O	O
-317/-288	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
present	NN	O	O
within	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
was	NN	O	O
still	NN	O	O
responsive	NN	O	O
to	NN	O	O
Tat	NN	O	B-protein
.	NN	O	O

A	NN	O	O
single	NN	O	B-DNA
point	NN	O	I-DNA
mutation	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
SRE	NN	O	B-DNA
completely	NN	O	O
abrogated	NN	O	O
the	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
tat/S	NN	O	B-DNA
.	NN	O	O

Exogenous	NN	O	O
recombinant	NN	O	B-protein
Tat	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
also	NN	O	O
able	NN	O	O
to	NN	O	O
upregulate	NN	O	O
c-fos	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
serum-activated	NN	O	O
Jurkat	NN	O	O
and	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
endogenous	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
c-Fos	NN	O	B-protein
protein	NN	O	O
synthesis	NN	O	O
in	NN	O	O
both	NN	O	O
serum-activated	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
primary	NN	O	B-cell_type
PBMC	NN	O	I-cell_type
.	NN	O	O

c-Fos	NN	O	B-protein
protein	NN	O	O
was	NN	O	O
shown	NN	O	O
essential	NN	O	O
for	NN	O	O
an	NN	O	O
optimal	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
by	NN	O	O
Tat	NN	O	B-protein
:	NN	O	O
incubation	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
antisense	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
sense	NN	O	O
,	NN	O	O
c-fos	NN	O	O
oligonucleotides	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
either	NN	O	O
the	NN	O	O
Tat	NN	O	B-protein
-enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
LTR-CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
or	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
gag	NN	O	B-protein
p24	NN	O	I-protein
in	NN	O	O
the	NN	O	O
culture	NN	O	O
supernatants	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
PBMC	NN	O	B-cell_type
acutely	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
upregulation	NN	O	O
mediated	NN	O	O
by	NN	O	O
Tat	NN	O	B-protein
might	NN	O	O
play	NN	O	O
a	NN	O	O
significant	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
viral	NN	O	O
gene	NN	O	O
transactivation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cytokine	NN	O	B-protein
signal	NN	O	I-protein
networks	NN	O	I-protein
and	NN	O	O
a	NN	O	O
new	NN	O	O
era	NN	O	O
in	NN	O	O
biomedical	NN	O	O
research	NN	O	O
.	NN	O	O

Elucidation	NN	O	O
of	NN	O	O
the	NN	O	O
biochemical	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
that	NN	O	O
regulate	NN	O	O
transcription	NN	O	O
and	NN	O	O
replication	NN	O	O
is	NN	O	O
the	NN	O	O
focus	NN	O	O
of	NN	O	O
attention	NN	O	O
in	NN	O	O
molecular	NN	O	O
biology	NN	O	O
.	NN	O	O

This	NN	O	O
research	NN	O	O
may	NN	O	O
make	NN	O	O
feasible	NN	O	O
manipulation	NN	O	O
of	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
would	NN	O	O
have	NN	O	O
profound	NN	O	O
implication	NN	O	O
in	NN	O	O
biomedical	NN	O	O
research	NN	O	O
on	NN	O	O
cell	NN	O	O
and	NN	O	O
gene	NN	O	O
therapy	NN	O	O
,	NN	O	O
and	NN	O	O
development	NN	O	O
of	NN	O	O
pharmaceutical	NN	O	O
products	NN	O	O
.	NN	O	O

Cytokines	NN	O	B-protein
control	NN	O	O
growth	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
death	NN	O	O
,	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
cells	NN	O	O
of	NN	O	O
lymphocytic	NN	O	B-cell_type
,	NN	O	I-cell_type
hemopoietic	NN	O	I-cell_type
systems	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
together	NN	O	O
with	NN	O	O
nerve	NN	O	B-cell_type
cells	NN	O	I-cell_type
provide	NN	O	O
a	NN	O	O
pertinent	NN	O	O
model	NN	O	O
to	NN	O	O
study	NN	O	O
intercellular	NN	O	O
communications	NN	O	O
and	NN	O	O
intercellular	NN	O	O
signal	NN	O	O
networks	NN	O	O
.	NN	O	O

This	NN	O	O
review	NN	O	O
outlines	NN	O	O
general	NN	O	O
features	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
and	NN	O	O
several	NN	O	O
aspects	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
networks	NN	O	I-protein
are	NN	O	O
discussed	NN	O	O
with	NN	O	O
emphasis	NN	O	O
on	NN	O	O
:	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
Th1	NN	O	O
and	NN	O	O
Th2-specific	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
their	NN	O	O
receptors	NN	O	O
in	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
hemopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
manipulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
networks	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
tissue	NN	O	B-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Role	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
,	NN	O	O
and	NN	O	O
Sp1	NN	O	B-protein
proteins	NN	O	O
in	NN	O	O
uninduced	NN	O	O
and	NN	O	O
lipopolysaccharide-induced	NN	O	O
expression	NN	O	O
.	NN	O	O

Tissue	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
TF	NN	O	O
)	NN	O	O
expression	NN	O	O
by	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
during	NN	O	O
sepsis	NN	O	O
initiates	NN	O	O
intravascular	NN	O	O
thrombosis	NN	O	O
.	NN	O	O

Bacterial	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
rapidly	NN	O	O
induces	NN	O	O
TF	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
TF	NN	O	I-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	O
factors	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
c-Rel/p65	NN	O	B-protein
,	NN	O	O
Egr-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Sp1	NN	O	B-protein
.	NN	O	O

NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
proteins	NN	O	I-protein
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
physically	NN	O	O
interact	NN	O	O
with	NN	O	O
both	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	I-protein
Sp1	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
uninduced	NN	O	O
and	NN	O	O
LPS-induced	NN	O	O
TF	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
THP-1	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Deletional	NN	O	O
analysis	NN	O	O
indicated	NN	O	O
that	NN	O	O
five	NN	O	O
Sp1	NN	O	B-DNA
sites	NN	O	I-DNA
mediated	NN	O	O
basal	NN	O	O
expression	NN	O	O
in	NN	O	O
uninduced	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
two	NN	O	O
AP-1	NN	O	B-DNA
sites	NN	O	I-DNA
bound	NN	O	O
c-Fos/c-Jun	NN	O	B-protein
heterodimers	NN	O	I-protein
in	NN	O	O
both	NN	O	O
unstimulated	NN	O	B-cell_type
and	NN	O	I-cell_type
LPS-stimulated	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Maximal	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
TF	NN	O	B-DNA
promoter	NN	O	I-DNA
required	NN	O	O
the	NN	O	O
two	NN	O	O
AP-1	NN	O	B-protein
sites	NN	O	O
and	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
LPS	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Disruption	NN	O	O
of	NN	O	O
the	NN	O	O
conserved	NN	O	O
spacing	NN	O	O
between	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
AP-1	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
abolished	NN	O	O
LPS	NN	O	O
induction	NN	O	O
.	NN	O	O

Replacement	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
AP-1	NN	O	B-DNA
sites	NN	O	I-DNA
with	NN	O	O
intrinsically	NN	O	O
bent	NN	O	O
DNA	NN	O	O
partially	NN	O	O
restored	NN	O	O
LPS	NN	O	O
induction	NN	O	O
,	NN	O	O
suggesting	NN	O	O
an	NN	O	O
additional	NN	O	O
structural	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

Synergistic	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
LPS	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
Drosophila	NN	O	B-cell_line
Schneider	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
coexpression	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p65	NN	O	B-protein
or	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
required	NN	O	O
the	NN	O	O
transactivation	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicated	NN	O	O
that	NN	O	O
c-Fos	NN	O	B-protein
/c-Jun	NN	O	B-protein
,	NN	O	O
c-Rel	NN	O	B-protein
/p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Sp1	NN	O	B-protein
regulate	NN	O	O
TF	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
growth	NN	O	O
hormone	NN	O	O
on	NN	O	O
TNF-alpha	NN	O	B-protein
secretion	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
translocation	NN	O	O
in	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Several	NN	O	O
studies	NN	O	O
have	NN	O	O
pointed	NN	O	O
to	NN	O	O
a	NN	O	O
link	NN	O	O
between	NN	O	O
immune	NN	O	O
and	NN	O	O
endocrine	NN	O	O
systems	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
regulatory	NN	O	O
function	NN	O	O
of	NN	O	O
GH	NN	O	O
on	NN	O	O
monocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
human	NN	O	O
THP-1	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
engineered	NN	O	O
by	NN	O	O
gene	NN	O	O
transfer	NN	O	O
to	NN	O	O
constitutively	NN	O	O
produce	NN	O	O
human	NN	O	O
growth	NN	O	O
hormone	NN	O	O
(	NN	O	O
hGH	NN	O	O
)	NN	O	O
,	NN	O	O
secreted	NN	O	O
depressed	NN	O	O
amounts	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
challenge	NN	O	O
by	NN	O	O
LPS	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
GH	NN	O	O
appears	NN	O	O
to	NN	O	O
occur	NN	O	O
in	NN	O	O
an	NN	O	O
autocrine	NN	O	O
fashion	NN	O	O
,	NN	O	O
since	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
TNF-alpha	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
constitutive	NN	O	O
GH	NN	O	O
production	NN	O	O
could	NN	O	O
be	NN	O	O
abolished	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
anti-hGH	NN	O	B-protein
mAb	NN	O	I-protein
.	NN	O	O

The	NN	O	O
GH-induced	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
using	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
by	NN	O	O
THP-1-hGH-transfected	NN	O	B-cell_line
cells	NN	O	I-cell_line
cultured	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
LPS	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
a	NN	O	O
selective	NN	O	O
pathway	NN	O	O
,	NN	O	O
since	NN	O	O
no	NN	O	O
inhibition	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
was	NN	O	O
observed	NN	O	O
when	NN	O	O
cells	NN	O	O
were	NN	O	O
cultured	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
PMA	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
LPS-stimulated	NN	O	B-cell_line
THP-1-hGH	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
.	NN	O	O

The	NN	O	O
capacity	NN	O	O
of	NN	O	O
GH	NN	O	O
to	NN	O	O
inhibit	NN	O	O
LPS-induced	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
without	NN	O	O
altering	NN	O	O
other	NN	O	O
pathways	NN	O	O
leading	NN	O	O
to	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
may	NN	O	O
be	NN	O	O
of	NN	O	O
potential	NN	O	O
relevance	NN	O	O
in	NN	O	O
septic	NN	O	O
shock	NN	O	O
,	NN	O	O
since	NN	O	O
GH	NN	O	O
is	NN	O	O
available	NN	O	O
for	NN	O	O
clinical	NN	O	O
use	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
lipopolysaccharide	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
fibroblast	NN	O	O
conditioned	NN	O	O
medium	NN	O	O
and	NN	O	O
exogenous	NN	O	O
PGE2	NN	O	O
.	NN	O	O

The	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
crucially	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
gene	NN	O	O
activation	NN	O	O
of	NN	O	O
several	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
including	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
fibroblast	NN	O	O
conditioned	NN	O	O
medium	NN	O	O
(	NN	O	O
FCM	NN	O	O
)	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
inhibit	NN	O	O
both	NN	O	O
TNF	NN	O	B-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
and	NN	O	O
protein	NN	O	O
release	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_line
blood-derived	NN	O	I-cell_line
human	NN	O	I-cell_line
monocytes	NN	O	I-cell_line
(	NN	O	O
PBM	NN	O	B-cell_line
)	NN	O	O
stimulated	NN	O	O
with	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
FCM	NN	O	O
on	NN	O	O
the	NN	O	O
LPS-induced	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
electrophoretic	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
FCM	NN	O	O
strongly	NN	O	O
inhibits	NN	O	O
the	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
PBM	NN	O	B-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
exogenous	NN	O	O
PGE2	NN	O	O
mimics	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
FCM	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
FCM	NN	O	O
produced	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
indomethacin	NN	O	O
does	NN	O	O
not	NN	O	O
inhibit	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
LPS	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
lend	NN	O	O
further	NN	O	O
support	NN	O	O
to	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
inflammatory	NN	O	O
and	NN	O	O
immune	NN	O	O
responses	NN	O	O
of	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
may	NN	O	O
be	NN	O	O
modulated	NN	O	O
at	NN	O	O
the	NN	O	O
molecular	NN	O	O
level	NN	O	O
by	NN	O	O
signals	NN	O	O
originating	NN	O	O
from	NN	O	O
tissue	NN	O	B-cell_type
structural	NN	O	I-cell_type
cells	NN	O	I-cell_type
such	NN	O	O
as	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-protein
neutrophil	NN	O	I-protein
elastase	NN	O	I-protein
proteolytically	NN	O	O
activates	NN	O	O
the	NN	O	O
platelet	NN	O	B-protein
integrin	NN	O	I-protein
alphaIIbbeta3	NN	O	B-protein
through	NN	O	O
cleavage	NN	O	O
of	NN	O	O
the	NN	O	O
carboxyl	NN	O	B-protein
terminus	NN	O	I-protein
of	NN	O	O
the	NN	O	O
alphaIIb	NN	O	B-protein
subunit	NN	O	I-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
.	NN	O	O

Involvement	NN	O	O
in	NN	O	O
the	NN	O	O
potentiation	NN	O	O
of	NN	O	O
platelet	NN	O	O
aggregation	NN	O	O
.	NN	O	O

Neutrophil	NN	O	B-protein
elastase	NN	O	I-protein
(	NN	O	O
NE	NN	O	B-protein
)	NN	O	O
and	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
are	NN	O	O
two	NN	O	O
serine	NN	O	B-protein
proteinases	NN	O	I-protein
released	NN	O	O
concomitantly	NN	O	O
by	NN	O	O
stimulated	NN	O	O
polymorphonuclear	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
while	NN	O	O
NE	NN	O	B-protein
by	NN	O	O
itself	NN	O	O
does	NN	O	O
not	NN	O	O
activate	NN	O	O
human	NN	O	B-cell_type
platelets	NN	O	I-cell_type
,	NN	O	O
it	NN	O	O
strongly	NN	O	O
enhances	NN	O	O
the	NN	O	O
weak	NN	O	O
aggregation	NN	O	O
induced	NN	O	O
by	NN	O	O
a	NN	O	O
threshold	NN	O	O
concentration	NN	O	O
of	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
(	NN	O	O
threshold	NN	O	O
of	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
)	NN	O	O
(	NN	O	O
Renesto	NN	O	O
,	NN	O	O
P.	NN	O	O
,	NN	O	O
and	NN	O	O
Chignard	NN	O	O
,	NN	O	O
M.	NN	O	O
(	NN	O	O
1993	NN	O	O
)	NN	O	O
Blood	NN	O	O
82	NN	O	O
,	NN	O	O
139-144	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
delineate	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
this	NN	O	O
potentiation	NN	O	O
process	NN	O	O
.	NN	O	O

Two	NN	O	O
main	NN	O	O
pieces	NN	O	O
of	NN	O	O
data	NN	O	O
prompted	NN	O	O
us	NN	O	O
to	NN	O	O
focus	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
platelet	NN	O	B-protein
fibrinogen	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
the	NN	O	O
alphaIIbbeta3	NN	O	B-protein
integrin	NN	O	I-protein
.	NN	O	O

First	NN	O	O
,	NN	O	O
previous	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
this	NN	O	O
integrin	NN	O	B-protein
to	NN	O	O
be	NN	O	O
particularly	NN	O	O
prone	NN	O	O
to	NN	O	O
proteolytic	NN	O	O
regulation	NN	O	O
of	NN	O	O
its	NN	O	O
function	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
potentiating	NN	O	O
activity	NN	O	O
of	NN	O	O
NE	NN	O	B-protein
on	NN	O	O
the	NN	O	O
threshold	NN	O	O
of	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
-induced	NN	O	O
platelet	NN	O	O
aggregation	NN	O	O
was	NN	O	O
strictly	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
fibrinogen	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
analysis	NN	O	O
,	NN	O	O
NE	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
trigger	NN	O	O
a	NN	O	O
time-dependent	NN	O	O
binding	NN	O	O
of	NN	O	O
PAC-1	NN	O	B-protein
and	NN	O	O
AP-5	NN	O	B-protein
,	NN	O	O
two	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
specific	NN	O	O
for	NN	O	O
the	NN	O	O
activated	NN	O	O
and	NN	O	O
ligand-occupied	NN	O	O
conformers	NN	O	O
of	NN	O	O
alphaIIbbeta3	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
potentiated	NN	O	O
aggregation	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
result	NN	O	O
from	NN	O	O
an	NN	O	O
increased	NN	O	O
capacity	NN	O	O
of	NN	O	O
platelets	NN	O	O
to	NN	O	O
bind	NN	O	O
fibrinogen	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
NE	NN	O	B-protein
and	NN	O	O
threshold	NN	O	O
of	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
increased	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
PAC-1	NN	O	B-protein
approximately	NN	O	O
5.5-fold	NN	O	O
over	NN	O	O
basal	NN	O	O
values	NN	O	O
measured	NN	O	O
on	NN	O	O
nontreated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
this	NN	O	O
binding	NN	O	O
raised	NN	O	O
only	NN	O	O
by	NN	O	O
approximately	NN	O	O
3-fold	NN	O	O
in	NN	O	O
threshold	NN	O	O
of	NN	O	O
cathepsin	NN	O	B-cell_type
G-stimulated	NN	O	I-cell_type
platelets	NN	O	I-cell_type
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
phosphatidic	NN	O	O
acid	NN	O	O
accumulation	NN	O	O
,	NN	O	O
pleckstrin	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
and	NN	O	O
calcium	NN	O	O
mobilization	NN	O	O
produced	NN	O	O
by	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
NE	NN	O	B-protein
and	NN	O	O
threshold	NN	O	O
of	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
were	NN	O	O
not	NN	O	O
significantly	NN	O	O
different	NN	O	O
from	NN	O	O
those	NN	O	O
measured	NN	O	O
with	NN	O	O
threshold	NN	O	O
of	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
alone	NN	O	O
(	NN	O	O
p	NN	O	O
>	NN	O	O
0.05	NN	O	O
)	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
phospholipase	NN	O	B-protein
C	NN	O	I-protein
/protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
pathway	NN	O	O
is	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
potentiation	NN	O	O
of	NN	O	O
aggregation	NN	O	O
.	NN	O	O

The	NN	O	O
foregoing	NN	O	O
data	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
requirement	NN	O	O
of	NN	O	O
catalytically	NN	O	O
active	NN	O	O
NE	NN	O	B-protein
to	NN	O	O
trigger	NN	O	O
alphaIIbbeta3	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
potentiate	NN	O	O
threshold	NN	O	O
of	NN	O	O
cathepsin	NN	O	B-protein
G	NN	O	I-protein
-initiated	NN	O	O
platelet	NN	O	O
aggregation	NN	O	O
,	NN	O	O
led	NN	O	O
us	NN	O	O
to	NN	O	O
examine	NN	O	O
whether	NN	O	O
the	NN	O	O
structure	NN	O	O
of	NN	O	O
this	NN	O	O
integrin	NN	O	B-protein
was	NN	O	O
affected	NN	O	O
by	NN	O	O
NE	NN	O	B-protein
.	NN	O	O

Immunoblot	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
a	NN	O	O
limited	NN	O	O
proteolysis	NN	O	O
of	NN	O	O
the	NN	O	O
carboxyl	NN	O	B-protein
terminus	NN	O	I-protein
of	NN	O	O
the	NN	O	O
alphaIIb	NN	O	B-protein
subunit	NN	O	I-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
alphaIIbH	NN	O	B-protein
)	NN	O	O
,	NN	O	O
as	NN	O	O
judged	NN	O	O
by	NN	O	O
the	NN	O	O
disappearance	NN	O	O
of	NN	O	O
the	NN	O	O
epitope	NN	O	O
for	NN	O	O
the	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
PMI-1	NN	O	I-protein
.	NN	O	O

Mass	NN	O	O
spectrometry	NN	O	O
studies	NN	O	O
performed	NN	O	O
on	NN	O	O
a	NN	O	O
synthetic	NN	O	O
peptide	NN	O	O
mapping	NN	O	O
over	NN	O	O
the	NN	O	O
cleavage	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
alphaIIbH	NN	O	B-protein
predicted	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
proteolysis	NN	O	O
as	NN	O	O
located	NN	O	O
between	NN	O	O
Val837	NN	O	O
and	NN	O	O
Asp838	NN	O	O
.	NN	O	O

Treatment	NN	O	O
by	NN	O	O
NE	NN	O	B-protein
of	NN	O	O
ATP-depleted	NN	O	B-cell_type
platelets	NN	O	I-cell_type
or	NN	O	O
Chinese	NN	O	B-cell_line
hamster	NN	O	I-cell_line
ovary	NN	O	I-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
human	NN	O	B-protein
recombinant	NN	O	I-protein
alphaIIbbeta3	NN	O	I-protein
clearly	NN	O	O
established	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
integrin	NN	O	B-protein
was	NN	O	O
independent	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
and	NN	O	O
was	NN	O	O
concomitant	NN	O	O
with	NN	O	O
the	NN	O	O
proteolysis	NN	O	O
of	NN	O	O
alphaIIbH	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

AP-1	NN	O	B-protein
derived	NN	O	O
from	NN	O	O
mature	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
astrocytes	NN	O	B-cell_type
preferentially	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
promoter	NN	O	I-DNA
central	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
interacting	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
lymphotropic	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
I	NN	O	I-DNA
(	NN	O	I-DNA
HTLV-I	NN	O	I-DNA
)	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
is	NN	O	O
essential	NN	O	O
to	NN	O	O
dissecting	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
viral	NN	O	O
transcription	NN	O	O
that	NN	O	O
may	NN	O	O
be	NN	O	O
pertinent	NN	O	O
to	NN	O	O
the	NN	O	O
oncogenic	NN	O	O
and	NN	O	O
neuropathogenic	NN	O	O
processes	NN	O	O
associated	NN	O	O
with	NN	O	O
HTLV-I	NN	O	O
infection	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
immune	NN	O	O
and	NN	O	O
nervous	NN	O	O
systems	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
(	NN	O	O
EMS	NN	O	O
)	NN	O	O
analyses	NN	O	O
utilizing	NN	O	O
oligonucleotides	NN	O	O
homologous	NN	O	O
to	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeat	NN	O	I-DNA
elements	NN	O	I-DNA
reacted	NN	O	O
with	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
derived	NN	O	O
from	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
of	NN	O	O
lymphocytic	NN	O	B-cell_type
,	NN	O	I-cell_type
monocytic	NN	O	I-cell_type
,	NN	O	I-cell_type
neuronal	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
glial	NN	O	I-cell_type
cell	NN	O	I-cell_type
origin	NN	O	I-cell_type
have	NN	O	O
demonstrated	NN	O	O
differential	NN	O	O
binding	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
three	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeats	NN	O	I-DNA
(	NN	O	O
1-4	NN	O	O
)	NN	O	O
.	NN	O	O

ATF/CREB	NN	O	B-protein
and	NN	O	O
Sp	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
interacted	NN	O	O
with	NN	O	O
the	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeats	NN	O	I-DNA
to	NN	O	O
form	NN	O	O
DNA-protein	NN	O	B-protein
complexes	NN	O	I-protein
common	NN	O	O
to	NN	O	O
all	NN	O	O
cell	NN	O	O
types	NN	O	O
examined	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
unique	NN	O	O
DNA-protein	NN	O	B-protein
complex	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
when	NN	O	O
the	NN	O	O
promoter	NN	O	O
central	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeat	NN	O	I-DNA
was	NN	O	O
reacted	NN	O	O
with	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
derived	NN	O	O
from	NN	O	O
either	NN	O	O
the	NN	O	O
U-373	NN	O	B-cell_line
MG	NN	O	I-cell_line
glioblastoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
or	NN	O	O
the	NN	O	O
THP-1	NN	O	B-cell_line
mature	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Based	NN	O	O
on	NN	O	O
nucleotide	NN	O	O
sequence	NN	O	O
requirements	NN	O	O
and	NN	O	O
immunoreactivity	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
this	NN	O	O
DNA-protein	NN	O	B-protein
complex	NN	O	I-protein
is	NN	O	O
comprised	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
components	NN	O	O
,	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

HIV	NN	O	O
does	NN	O	O
not	NN	O	O
replicate	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
CD4	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
CD3/CD28	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
tropic	NN	O	O
strain	NN	O	O
of	NN	O	O
HIV	NN	O	O
,	NN	O	O
LAI	NN	O	O
,	NN	O	O
does	NN	O	O
not	NN	O	O
replicate	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
CD4	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
cross-linking	NN	O	O
CD3	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
LAI	NN	O	O
replicates	NN	O	O
well	NN	O	O
in	NN	O	O
memory	NN	O	B-cell_type
CD4	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
way	NN	O	O
.	NN	O	O

Unlike	NN	O	O
this	NN	O	O
physiologically	NN	O	O
relevant	NN	O	O
stimulation	NN	O	O
,	NN	O	O
PHA	NN	O	B-protein
stimulates	NN	O	O
productive	NN	O	O
LAI	NN	O	O
replication	NN	O	O
in	NN	O	O
both	NN	O	O
naive	NN	O	B-cell_type
and	NN	O	I-cell_type
memory	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
studies	NN	O	O
were	NN	O	O
conducted	NN	O	O
with	NN	O	O
highly	NN	O	O
purified	NN	O	O
(	NN	O	O
FACS-isolated	NN	O	O
)	NN	O	O
subsets	NN	O	O
of	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
identified	NN	O	O
by	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
CD45RA	NN	O	B-protein
and	NN	O	O
CD62L	NN	O	B-protein
.	NN	O	O

Remixing	NN	O	O
of	NN	O	O
purified	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
showed	NN	O	O
that	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
do	NN	O	O
not	NN	O	O
suppress	NN	O	O
LAI	NN	O	O
replication	NN	O	O
in	NN	O	O
memory	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
memory	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
do	NN	O	O
not	NN	O	O
restore	NN	O	O
LAI	NN	O	O
expression	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
suppression	NN	O	O
of	NN	O	O
productive	NN	O	O
LAI	NN	O	O
replication	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
differential	NN	O	O
expression	NN	O	O
of	NN	O	O
viral	NN	O	B-protein
coreceptors	NN	O	I-protein
,	NN	O	O
nor	NN	O	O
is	NN	O	O
it	NN	O	O
due	NN	O	O
to	NN	O	O
inhibition	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
important	NN	O	O
HIV	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
and	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
.	NN	O	O

The	NN	O	O
inherent	NN	O	O
resistance	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
productive	NN	O	O
HIV	NN	O	O
infection	NN	O	O
,	NN	O	O
coupled	NN	O	O
with	NN	O	O
their	NN	O	O
proliferative	NN	O	O
advantage	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
here	NN	O	O
,	NN	O	O
provides	NN	O	O
a	NN	O	O
sound	NN	O	O
basis	NN	O	O
for	NN	O	O
proposed	NN	O	O
clinical	NN	O	O
therapies	NN	O	O
using	NN	O	O
ex	NN	O	O
vivo	NN	O	O
expansion	NN	O	O
and	NN	O	O
reinfusion	NN	O	O
of	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
HIV-infected	NN	O	O
adults	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
PMLRARalpha	NN	O	B-DNA
transgene	NN	O	I-DNA
initiates	NN	O	O
murine	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

The	NN	O	O
malignant	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
contain	NN	O	O
a	NN	O	O
reciprocal	NN	O	O
chromosomal	NN	O	O
translocation	NN	O	O
that	NN	O	O
fuses	NN	O	O
the	NN	O	O
promyelocytic	NN	O	B-DNA
leukemia	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
PML	NN	O	O
)	NN	O	O
with	NN	O	O
the	NN	O	O
retinoic	NN	O	B-DNA
acid	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
the	NN	O	O
chimera	NN	O	B-protein
PMLRAR	NN	O	I-protein
alpha	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
leukemogenesis	NN	O	O
,	NN	O	O
we	NN	O	O
expressed	NN	O	O
a	NN	O	O
PMLRAR	NN	O	B-DNA
alpha	NN	O	I-DNA
cDNA	NN	O	I-DNA
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

PMLRAR	NN	O	O
alpha	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
exhibited	NN	O	O
impaired	NN	O	O
neutrophil	NN	O	O
maturation	NN	O	O
early	NN	O	O
in	NN	O	O
life	NN	O	O
,	NN	O	O
which	NN	O	O
progressed	NN	O	O
at	NN	O	O
a	NN	O	O
low	NN	O	O
frequency	NN	O	O
over	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
several	NN	O	O
months	NN	O	O
to	NN	O	O
overt	NN	O	O
APL	NN	O	O
.	NN	O	O

Both	NN	O	O
the	NN	O	O
preleukemic	NN	O	O
state	NN	O	O
and	NN	O	O
the	NN	O	O
leukemia	NN	O	O
could	NN	O	O
be	NN	O	O
transplanted	NN	O	O
to	NN	O	O
nontransgenic	NN	O	O
mice	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
transplanted	NN	O	O
preleukemia	NN	O	O
could	NN	O	O
progress	NN	O	O
to	NN	O	O
APL	NN	O	O
.	NN	O	O

The	NN	O	O
APL	NN	O	O
recapitulated	NN	O	O
features	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
disease	NN	O	O
,	NN	O	O
including	NN	O	O
a	NN	O	O
response	NN	O	O
to	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
.	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
caused	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
differentiate	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
eliciting	NN	O	O
remissions	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
preleukemic	NN	O	O
state	NN	O	O
and	NN	O	O
APL	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
PMLRAR	NN	O	B-protein
alpha	NN	O	I-protein
impairs	NN	O	O
neutrophil	NN	O	O
differentiation	NN	O	O
and	NN	O	O
initiates	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
APL	NN	O	O
.	NN	O	O

The	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
described	NN	O	O
here	NN	O	O
provide	NN	O	O
an	NN	O	O
apparently	NN	O	O
accurate	NN	O	O
model	NN	O	O
for	NN	O	O
human	NN	O	O
APL	NN	O	O
that	NN	O	O
includes	NN	O	O
clear	NN	O	O
evidence	NN	O	O
of	NN	O	O
tumor	NN	O	O
progression	NN	O	O
.	NN	O	O

The	NN	O	O
model	NN	O	O
should	NN	O	O
be	NN	O	O
useful	NN	O	O
for	NN	O	O
exploring	NN	O	O
the	NN	O	O
molecular	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
APL	NN	O	O
and	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
the	NN	O	O
therapeutic	NN	O	O
response	NN	O	O
to	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
for	NN	O	O
preclinical	NN	O	O
studies	NN	O	O
of	NN	O	O
therapeutic	NN	O	O
regimens	NN	O	O
.	NN	O	O

-DOCSTART-	O

DNA	NN	O	O
methylation	NN	O	O
changes	NN	O	O
in	NN	O	O
hematologic	NN	O	O
malignancies	NN	O	O
:	NN	O	O
biologic	NN	O	O
and	NN	O	O
clinical	NN	O	O
implications	NN	O	O
.	NN	O	O

DNA	NN	O	O
methylation	NN	O	O
changes	NN	O	O
are	NN	O	O
among	NN	O	O
the	NN	O	O
most	NN	O	O
common	NN	O	O
detectable	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
human	NN	O	O
neoplasia	NN	O	O
.	NN	O	O

Hypermethylation	NN	O	O
within	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
selected	NN	O	O
genes	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
especially	NN	O	O
common	NN	O	O
in	NN	O	O
all	NN	O	O
types	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
neoplasms	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
is	NN	O	O
usually	NN	O	O
associated	NN	O	O
with	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
involved	NN	O	O
gene	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
.	NN	O	O

Such	NN	O	O
hypermethylation-associated	NN	O	O
silencing	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
for	NN	O	O
several	NN	O	O
genes	NN	O	O
regulating	NN	O	O
the	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
including	NN	O	O
the	NN	O	O
estrogen	NN	O	B-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
ER	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
P15	NN	O	O
,	NN	O	O
P16	NN	O	O
and	NN	O	O
others	NN	O	O
.	NN	O	O

Hypermethylation	NN	O	O
within	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
some	NN	O	O
genes	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
early	NN	O	O
event	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
neoplasia	NN	O	O
(	NN	O	O
ER	NN	O	B-DNA
,	NN	O	O
P15	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
while	NN	O	O
other	NN	O	O
genes	NN	O	O
seem	NN	O	O
to	NN	O	O
become	NN	O	O
methylated	NN	O	O
during	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
leukemias	NN	O	O
(	NN	O	O
HIC1	NN	O	O
,	NN	O	O
c-abl	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
high	NN	O	O
prevalence	NN	O	O
of	NN	O	O
promoter	NN	O	O
methylation	NN	O	O
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
molecular	NN	O	O
abnormality	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
monitor	NN	O	O
disease	NN	O	O
activity	NN	O	O
during	NN	O	O
therapy	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
new	NN	O	O
technology	NN	O	O
allows	NN	O	O
the	NN	O	O
sensitive	NN	O	O
identification	NN	O	O
of	NN	O	O
gene	NN	O	O
hypermethylation	NN	O	O
in	NN	O	O
a	NN	O	O
background	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
possible	NN	O	O
new	NN	O	O
strategies	NN	O	O
for	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
minimal	NN	O	O
residual	NN	O	O
disease	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
reactivation	NN	O	O
of	NN	O	O
tumor-suppressor	NN	O	O
gene	NN	O	O
expression	NN	O	O
through	NN	O	O
pharmacologic	NN	O	O
inhibition	NN	O	O
of	NN	O	O
DNA	NN	O	B-protein
methyltransferase	NN	O	I-protein
and	NN	O	O
resultant	NN	O	O
DNA	NN	O	O
demethylation	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
promising	NN	O	O
new	NN	O	O
avenue	NN	O	O
of	NN	O	O
therapy	NN	O	O
in	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

Redox	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
pathway	NN	O	O
during	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
an	NN	O	O
obligatory	NN	O	O
requirement	NN	O	O
for	NN	O	O
intracellular	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
generation	NN	O	O
during	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
have	NN	O	O
proposed	NN	O	O
that	NN	O	O
intracellular	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
may	NN	O	O
act	NN	O	O
as	NN	O	O
signalling	NN	O	O
agents	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
certain	NN	O	O
cellular	NN	O	O
processes	NN	O	O
,	NN	O	O
for	NN	O	O
example	NN	O	O
,	NN	O	O
during	NN	O	O
cell	NN	O	O
cycle	NN	O	O
entry	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
been	NN	O	O
interested	NN	O	O
to	NN	O	O
determine	NN	O	O
which	NN	O	O
,	NN	O	O
if	NN	O	O
any	NN	O	O
,	NN	O	O
cell	NN	O	O
cycle	NN	O	O
entry	NN	O	O
events	NN	O	O
are	NN	O	O
affected	NN	O	O
by	NN	O	O
oxidative	NN	O	O
signalling	NN	O	O
.	NN	O	O

In	NN	O	O
earlier	NN	O	O
studies	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
as	NN	O	O
molecular	NN	O	O
targets	NN	O	O
for	NN	O	O
oxidative	NN	O	O
signalling	NN	O	O
processes	NN	O	O
during	NN	O	O
cell	NN	O	O
cycle	NN	O	O
entry	NN	O	O
,	NN	O	O
and	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
oxidative	NN	O	O
signalling	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
early	NN	O	O
changes	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
the	NN	O	O
G0	NN	O	O
to	NN	O	O
G1	NN	O	O
phase	NN	O	O
transition	NN	O	O
.	NN	O	O

To	NN	O	O
extend	NN	O	O
these	NN	O	O
initial	NN	O	O
observations	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
antioxidant	NN	O	O
treatment	NN	O	O
on	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
erk1	NN	O	B-protein
and	NN	O	O
erk2	NN	O	B-protein
,	NN	O	O
as	NN	O	O
members	NN	O	O
of	NN	O	O
a	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
known	NN	O	O
to	NN	O	O
directly	NN	O	O
regulate	NN	O	O
transcription	NN	O	O
factor	NN	O	O
function	NN	O	O
.	NN	O	O

Using	NN	O	O
as	NN	O	O
a	NN	O	O
probe	NN	O	O
cysteamine	NN	O	O
,	NN	O	O
an	NN	O	O
aminothiol	NN	O	O
compound	NN	O	O
with	NN	O	O
both	NN	O	O
antioxidant	NN	O	O
and	NN	O	O
antiproliferative	NN	O	O
activity	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
erk2	NN	O	B-protein
,	NN	O	O
a	NN	O	O
key	NN	O	O
element	NN	O	O
of	NN	O	O
the	NN	O	O
MAP	NN	O	O
kinase	NN	O	O
pathway	NN	O	O
,	NN	O	O
as	NN	O	O
being	NN	O	O
responsive	NN	O	O
to	NN	O	O
oxidative	NN	O	O
signalling	NN	O	O
during	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
provide	NN	O	O
further	NN	O	O
evidence	NN	O	O
to	NN	O	O
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
intracellular	NN	O	O
oxidant	NN	O	O
generation	NN	O	O
as	NN	O	O
a	NN	O	O
regulatory	NN	O	O
mechanism	NN	O	O
during	NN	O	O
cell	NN	O	O
cycle	NN	O	O
entry	NN	O	O
,	NN	O	O
and	NN	O	O
establish	NN	O	O
a	NN	O	O
link	NN	O	O
between	NN	O	O
oxidative	NN	O	O
signalling	NN	O	O
and	NN	O	O
other	NN	O	O
aspects	NN	O	O
of	NN	O	O
the	NN	O	O
intracellular	NN	O	O
signalling	NN	O	O
network	NN	O	O
that	NN	O	O
is	NN	O	O
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Physical	NN	O	O
and	NN	O	O
functional	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
lymphotropic	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
Tax1	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
the	NN	O	O
CCAAT	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
NF-Y	NN	O	B-protein
.	NN	O	O

Tax1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
potent	NN	O	O
activator	NN	O	O
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
lymphotropic	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
transcription	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
modulate	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Tax1	NN	O	B-protein
does	NN	O	O
not	NN	O	O
bind	NN	O	O
DNA	NN	O	O
directly	NN	O	O
but	NN	O	O
regulates	NN	O	O
transcription	NN	O	O
through	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
with	NN	O	O
sequence-specific	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
the	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
system	NN	O	O
to	NN	O	O
screen	NN	O	O
for	NN	O	O
proteins	NN	O	O
which	NN	O	O
interact	NN	O	O
with	NN	O	O
Tax1	NN	O	B-protein
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
the	NN	O	O
B	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
CCAAT	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
NF-Y	NN	O	B-protein
from	NN	O	O
a	NN	O	O
HeLa	NN	O	B-DNA
cDNA	NN	O	I-DNA
library	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
interaction	NN	O	O
of	NN	O	O
Tax1	NN	O	B-protein
with	NN	O	O
NF-YB	NN	O	B-protein
was	NN	O	O
specific	NN	O	O
in	NN	O	O
that	NN	O	O
NF-YB	NN	O	B-protein
did	NN	O	O
not	NN	O	O
interact	NN	O	O
with	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
human	NN	O	B-protein
immunodeficiency	NN	O	I-protein
virus	NN	O	I-protein
Tat	NN	O	I-protein
,	NN	O	O
human	NN	O	B-protein
papillomavirus	NN	O	I-protein
E6	NN	O	I-protein
,	NN	O	O
and	NN	O	O
Bicoid	NN	O	B-protein
,	NN	O	O
or	NN	O	O
with	NN	O	O
the	NN	O	O
M7	NN	O	B-protein
(	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
29CP-AS	NN	O	I-protein
)	NN	O	I-protein
Tax1	NN	O	I-protein
mutant	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
NF-YB	NN	O	B-protein
did	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
Tax1	NN	O	I-protein
mutants	NN	O	I-protein
M22	NN	O	B-protein
(	NN	O	O
130TL-AS	NN	O	B-protein
)	NN	O	O
and	NN	O	O
M47	NN	O	B-protein
(	NN	O	O
319LL-RS	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
vitro-translated	NN	O	O
NF-YB	NN	O	B-protein
specifically	NN	O	O
bound	NN	O	O
to	NN	O	O
a	NN	O	O
glutathione	NN	O	B-protein
S-transferase-Tax1	NN	O	I-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Further	NN	O	O
,	NN	O	O
Tax1	NN	O	B-protein
coimmunoprecipitated	NN	O	O
with	NN	O	O
NF-Y	NN	O	B-protein
from	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
HTLV-1-transformed	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
providing	NN	O	O
evidence	NN	O	O
for	NN	O	O
in	NN	O	O
vivo	NN	O	O
interaction	NN	O	O
of	NN	O	O
Tax1	NN	O	B-protein
and	NN	O	O
NF-YB	NN	O	B-protein
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Tax1	NN	O	B-protein
specifically	NN	O	O
activated	NN	O	O
the	NN	O	O
NF-Y-responsive	NN	O	B-DNA
DQbeta	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
minimal	NN	O	B-DNA
promoter	NN	O	I-DNA
which	NN	O	O
contains	NN	O	O
only	NN	O	O
the	NN	O	O
Y-box	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
Y-box	NN	O	B-DNA
element	NN	O	I-DNA
alone	NN	O	O
abrogated	NN	O	O
Tax1	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
Tax1	NN	O	B-protein
interacts	NN	O	O
with	NN	O	O
NF-Y	NN	O	B-protein
through	NN	O	O
the	NN	O	O
B	NN	O	B-protein
subunit	NN	O	I-protein
and	NN	O	O
that	NN	O	O
this	NN	O	O
interaction	NN	O	O
results	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Through	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
and	NN	O	O
other	NN	O	O
NF-Y	NN	O	B-protein
driven	NN	O	O
promoters	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
Tax1	NN	O	B-protein
-NF-Y	NN	O	B-protein
interaction	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
causing	NN	O	O
cellular	NN	O	O
transformation	NN	O	O
and	NN	O	O
HTLV-1	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

High	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
in	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
and	NN	O	O
human	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
virus	NN	O	I-cell_type
type	NN	O	I-cell_type
I-infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
possible	NN	O	O
enhancement	NN	O	O
of	NN	O	O
DNA-binding	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
by	NN	O	O
the	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
I	NN	O	I-protein
transactivating	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
E2F	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
cellular	NN	O	B-DNA
promoters	NN	O	I-DNA
of	NN	O	O
certain	NN	O	O
growth-	NN	O	B-DNA
and	NN	O	I-DNA
cell	NN	O	I-DNA
cycle-controlling	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
forms	NN	O	O
distinct	NN	O	O
heteromeric	NN	O	B-protein
complexes	NN	O	I-protein
with	NN	O	O
other	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

It	NN	O	O
has	NN	O	O
therefore	NN	O	O
been	NN	O	O
proposed	NN	O	O
that	NN	O	O
E2F	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
control	NN	O	O
.	NN	O	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
etiological	NN	O	O
agent	NN	O	O
of	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ATL	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
by	NN	O	O
mobility-shift	NN	O	O
assay	NN	O	O
that	NN	O	O
E2F-containing	NN	O	B-protein
DNA-binding	NN	O	I-protein
complexes	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
in	NN	O	O
HTLV-I-infected	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
ATL	NN	O	O
patients	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
an	NN	O	O
uninfected	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
Jurkat	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
normal	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
encoded	NN	O	O
by	NN	O	O
HTLV-I	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
transcription	NN	O	O
activator	NN	O	O
of	NN	O	O
viral	NN	O	O
and	NN	O	O
several	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
can	NN	O	O
induce	NN	O	O
the	NN	O	O
E2F-containing	NN	O	B-protein
DNA-binding	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Tax	NN	O	B-protein
,	NN	O	O
through	NN	O	O
enhancement	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
,	NN	O	O
may	NN	O	O
be	NN	O	O
capable	NN	O	O
of	NN	O	O
regulating	NN	O	O
cellular	NN	O	O
gene	NN	O	O
expression	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and	NN	O	O
transformation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
ATL	NN	O	O
.	NN	O	O

This	NN	O	O
activity	NN	O	O
may	NN	O	O
be	NN	O	O
relevant	NN	O	O
to	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
whereby	NN	O	O
HTLV-I	NN	O	O
which	NN	O	O
does	NN	O	O
not	NN	O	O
contain	NN	O	O
oncogenes	NN	O	B-DNA
induces	NN	O	O
neoplasia	NN	O	O
.	NN	O	O

-DOCSTART-	O

Dissociation	NN	O	O
of	NN	O	O
the	NN	O	O
Jak	NN	O	B-protein
kinase	NN	O	I-protein
pathway	NN	O	O
from	NN	O	O
G-CSF	NN	O	B-protein
receptor	NN	O	I-protein
signaling	NN	O	O
in	NN	O	O
neutrophils	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
G-CSFR	NN	O	B-protein
)	NN	O	O
induces	NN	O	O
rapid	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
multiple	NN	O	O
intracellular	NN	O	B-protein
substrates	NN	O	I-protein
in	NN	O	O
proliferating	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
nonproliferating	NN	O	B-cell_type
,	NN	O	I-cell_type
terminally	NN	O	I-cell_type
differentiated	NN	O	I-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
kinases	NN	O	O
that	NN	O	O
couple	NN	O	O
ligand	NN	O	O
binding	NN	O	O
to	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
substrates	NN	O	I-protein
by	NN	O	O
the	NN	O	O
G-CSFR	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
specific	NN	O	O
functional	NN	O	O
programs	NN	O	O
are	NN	O	O
largely	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
early	NN	O	O
signaling	NN	O	O
events	NN	O	O
in	NN	O	O
proliferating	NN	O	B-cell_type
and	NN	O	O
terminally	NN	O	B-cell_type
differentiated	NN	O	I-cell_type
cells	NN	O	I-cell_type
following	NN	O	O
G-CSF	NN	O	B-protein
stimulation	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
identical	NN	O	O
signaling	NN	O	O
cascades	NN	O	O
are	NN	O	O
activated	NN	O	O
.	NN	O	O

In	NN	O	O
murine	NN	O	B-cell_line
Ba/F3	NN	O	I-cell_line
cells	NN	O	I-cell_line
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	B-protein
G-CSFR	NN	O	I-protein
and	NN	O	O
NFS-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
constitutively	NN	O	O
expressing	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
G-CSFR	NN	O	I-protein
,	NN	O	O
G-CSF	NN	O	B-protein
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
Jak1	NN	O	B-protein
,	NN	O	O
Jak2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Tyk2	NN	O	B-protein
.	NN	O	O

Tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
Stat1	NN	O	B-protein
was	NN	O	O
also	NN	O	O
detected	NN	O	O
following	NN	O	O
G-CSF	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
mitogenically	NN	O	B-protein
incompetent	NN	O	I-protein
human	NN	O	I-protein
G-CSFR	NN	O	I-protein
mutant	NN	O	I-protein
in	NN	O	O
which	NN	O	O
Pro639	NN	O	O
and	NN	O	O
Pro641	NN	O	O
were	NN	O	O
substituted	NN	O	O
by	NN	O	O
alanine	NN	O	O
,	NN	O	O
the	NN	O	O
box	NN	O	B-protein
1	NN	O	I-protein
PDP	NN	O	I-protein
motif	NN	O	I-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
Jak	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
G-CSFR	NN	O	B-protein
,	NN	O	O
and	NN	O	O
recruitment	NN	O	O
of	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Notably	NN	O	O
,	NN	O	O
no	NN	O	O
activation	NN	O	O
of	NN	O	O
Jak1	NN	O	B-protein
,	NN	O	O
Jak2	NN	O	B-protein
,	NN	O	O
Tyk2	NN	O	B-protein
,	NN	O	O
Stat1	NN	O	B-protein
,	NN	O	O
or	NN	O	O
Stat3	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
following	NN	O	O
G-CSF	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
detectable	NN	O	O
activation	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
recently	NN	O	O
cloned	NN	O	O
Jak3	NN	O	B-protein
kinase	NN	O	I-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
as	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
undergo	NN	O	O
terminal	NN	O	O
neutrophilic	NN	O	O
maturation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
a	NN	O	O
lack	NN	O	O
of	NN	O	O
involvement	NN	O	O
of	NN	O	O
Jak	NN	O	B-protein
kinases	NN	O	I-protein
in	NN	O	O
signaling	NN	O	O
by	NN	O	O
the	NN	O	O
G-CSFR	NN	O	B-protein
in	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
utilization	NN	O	O
of	NN	O	O
alternative	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
in	NN	O	O
proliferating	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
Jak	NN	O	B-protein
-Stat	NN	O	B-protein
pathway	NN	O	O
correlates	NN	O	O
with	NN	O	O
proliferative	NN	O	O
signaling	NN	O	O
by	NN	O	O
the	NN	O	O
G-CSFR	NN	O	B-protein
and	NN	O	O
requires	NN	O	O
the	NN	O	O
membrane-proximal	NN	O	B-protein
box	NN	O	I-protein
1	NN	O	I-protein
PXP	NN	O	I-protein
motif	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
conserved	NN	O	O
in	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Impaired	NN	O	O
induction	NN	O	O
of	NN	O	O
c-fos/c-jun	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
binding	NN	O	O
distinct	NN	O	O
c-fos/c-jun	NN	O	B-DNA
promoter	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
aging	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
factor	NN	O	I-protein
c-Fos/c-Jun	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
responsiveness	NN	O	O
and	NN	O	O
is	NN	O	O
often	NN	O	O
impaired	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
aging	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
aberrancies	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
c-fos/c-jun	NN	O	B-DNA
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
or	NN	O	O
protein	NN	O	O
level	NN	O	O
might	NN	O	O
underlie	NN	O	O
the	NN	O	O
age-associated	NN	O	O
impairments	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Whereas	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
young	NN	O	O
subjects	NN	O	O
stimulated	NN	O	O
with	NN	O	O
cross-linked	NN	O	B-protein
anti-CD3epsilon	NN	O	I-protein
mAb	NN	O	I-protein
OKT3	NN	O	B-protein
plus	NN	O	O
PMA	NN	O	O
or	NN	O	O
with	NN	O	O
the	NN	O	O
lectin	NN	O	B-protein
PHA	NN	O	B-protein
plus	NN	O	O
PMA	NN	O	O
demonstrated	NN	O	O
considerable	NN	O	O
increases	NN	O	O
in	NN	O	O
c-Fos	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
but	NN	O	O
not	NN	O	O
c-Jun	NN	O	B-protein
was	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
in	NN	O	O
stimulated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
certain	NN	O	O
elderly	NN	O	O
subjects	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
RNase	NN	O	B-protein
protection	NN	O	O
assays	NN	O	O
revealed	NN	O	O
that	NN	O	O
anti-CD3/	NN	O	O
PMA-stimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
a	NN	O	O
substantial	NN	O	O
proportion	NN	O	O
of	NN	O	O
elderly	NN	O	O
subjects	NN	O	O
exhibited	NN	O	O
decreased	NN	O	O
levels	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
and/or	NN	O	I-RNA
c-jun	NN	O	I-RNA
mRNA	NN	O	I-RNA
compared	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
young	NN	O	O
subjects	NN	O	O
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
recognizing	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
consensus	NN	O	I-DNA
TRE	NN	O	I-DNA
motif	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
c-jun	NN	O	I-DNA
TRE-like	NN	O	I-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
serum	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
SRE	NN	O	B-DNA
)	NN	O	O
were	NN	O	O
determined	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
and	NN	O	I-cell_type
stimulated	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
young	NN	O	O
subjects	NN	O	O
resulted	NN	O	O
in	NN	O	O
coordinated	NN	O	O
increases	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
complexes	NN	O	I-protein
binding	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
TRE	NN	O	I-DNA
,	NN	O	O
c-jun	NN	O	B-DNA
TRE	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
c-fos	NN	O	B-DNA
SRE	NN	O	I-DNA
DNA	NN	O	I-DNA
sequence	NN	O	I-DNA
motifs	NN	O	I-DNA
,	NN	O	O
age-related	NN	O	O
reductions	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
were	NN	O	O
accompanied	NN	O	O
by	NN	O	O
decreased	NN	O	O
levels	NN	O	O
of	NN	O	O
c-jun	NN	O	B-protein
TRE	NN	O	I-protein
and	NN	O	I-protein
c-fos	NN	O	I-protein
SRE	NN	O	I-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
complexes	NN	O	I-protein
binding	NN	O	O
the	NN	O	O
SRE	NN	O	B-DNA
motif	NN	O	I-DNA
induced	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
young	NN	O	O
and	NN	O	O
elderly	NN	O	O
subjects	NN	O	O
contained	NN	O	O
serum	NN	O	B-protein
response	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
Elk-1	NN	O	B-protein
pointing	NN	O	O
toward	NN	O	O
age-related	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
distinct	NN	O	O
from	NN	O	O
c-jun/AP-1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
underlying	NN	O	O
aberrancies	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
c-fos/c-jun	NN	O	B-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
their	NN	O	O
nuclear	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
age-related	NN	O	O
impairments	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Retinoic	NN	O	O
acid-induced	NN	O	O
modulation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
production	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	O
expression	NN	O	O
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
has	NN	O	O
important	NN	O	O
immune-modulating	NN	O	O
effects	NN	O	O
on	NN	O	O
both	NN	O	O
T	NN	O	O
and	NN	O	O
B	NN	O	O
cell	NN	O	O
function	NN	O	O
.	NN	O	O

Our	NN	O	O
laboratory	NN	O	O
has	NN	O	O
shown	NN	O	O
that	NN	O	O
RA	NN	O	O
can	NN	O	O
enhance	NN	O	O
in	NN	O	O
vitro	NN	O	O
polyclonal	NN	O	O
B	NN	O	O
cell	NN	O	O
immunoglobulin	NN	O	B-protein
(	NN	O	O
Ig	NN	O	O
)	NN	O	O
response	NN	O	O
.	NN	O	O

Investigating	NN	O	O
cytokines	NN	O	B-protein
known	NN	O	O
to	NN	O	O
affect	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
recently	NN	O	O
shown	NN	O	O
that	NN	O	O
IL-6	NN	O	B-protein
production	NN	O	O
is	NN	O	O
augmented	NN	O	O
by	NN	O	O
RA	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
immune	NN	O	O
modulating	NN	O	O
effects	NN	O	O
of	NN	O	O
RA	NN	O	O
on	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
another	NN	O	O
important	NN	O	O
cytokine	NN	O	B-protein
for	NN	O	O
B	NN	O	O
cell	NN	O	O
immunoglobulin	NN	O	B-protein
production	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
the	NN	O	O
RA	NN	O	B-protein
nuclear	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Purified	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
obtained	NN	O	O
from	NN	O	O
adenoidal	NN	O	O
tissues	NN	O	O
,	NN	O	O
and	NN	O	O
incubated	NN	O	O
with	NN	O	O
RA	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
or	NN	O	O
DMSO	NN	O	O
solvent/media	NN	O	O
control	NN	O	O
for	NN	O	O
0	NN	O	O
,	NN	O	O
6-8	NN	O	O
,	NN	O	O
and	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

Total	NN	O	O
mRNA	NN	O	B-RNA
was	NN	O	O
extracted	NN	O	O
from	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
using	NN	O	O
RT-PCR	NN	O	O
,	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
RA	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
)	NN	O	O
-	NN	O	O
alpha	NN	O	B-RNA
,	NN	O	O
beta	NN	O	B-RNA
,	NN	O	O
gamma	NN	O	B-RNA
mRNA	NN	O	I-RNA
were	NN	O	O
determined	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
RA	NN	O	O
on	NN	O	O
IL-2-alpha	NN	O	O
receptor	NN	O	O
expression	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
These	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
RA	NN	O	O
can	NN	O	O
augment	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
production	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
a	NN	O	O
possible	NN	O	O
paracrine	NN	O	O
effect	NN	O	O
on	NN	O	O
IL-2R-alpha	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

These	NN	O	O
changes	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
RAR-alpha	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
may	NN	O	O
be	NN	O	O
another	NN	O	O
important	NN	O	O
cytokine	NN	O	B-protein
modulated	NN	O	O
by	NN	O	O
RA	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
an	NN	O	O
SAF-like	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
serum	NN	O	B-DNA
amyloid	NN	O	I-DNA
A	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
lipopolysaccharide	NN	O	O
.	NN	O	O

Serum	NN	O	B-protein
amyloid	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	O
SAA	NN	O	B-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
linked	NN	O	O
to	NN	O	O
atherosclerosis	NN	O	O
because	NN	O	O
of	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
remodel	NN	O	O
high-density	NN	O	O
lipoprotein	NN	O	O
by	NN	O	O
the	NN	O	O
depletion	NN	O	O
of	NN	O	O
apolipoprotein	NN	O	O
A1	NN	O	O
,	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
bind	NN	O	O
cholesterol	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
presence	NN	O	O
in	NN	O	O
the	NN	O	O
atherosclerotic	NN	O	O
plaques	NN	O	O
of	NN	O	O
coronary	NN	O	O
and	NN	O	O
carotid	NN	O	O
arteries	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
induction	NN	O	O
mechanism	NN	O	O
of	NN	O	O
SAA	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
THP-1	NN	O	B-cell_line
monocyte/macrophage	NN	O	I-cell_line
cells	NN	O	I-cell_line
which	NN	O	O
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
atherosclerotic	NN	O	O
fatty	NN	O	O
streak	NN	O	O
and	NN	O	O
plaque	NN	O	O
formation	NN	O	O
.	NN	O	O

We	NN	O	O
and	NN	O	O
others	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
SAA	NN	O	B-protein
gene	NN	O	O
is	NN	O	O
induced	NN	O	O
in	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

By	NN	O	O
promoter	NN	O	O
function	NN	O	O
analysis	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
SAA	NN	O	B-DNA
promoter	NN	O	I-DNA
sequence	NN	O	I-DNA
between	NN	O	O
-280	NN	O	O
and	NN	O	O
-226	NN	O	O
can	NN	O	O
confer	NN	O	O
LPS	NN	O	O
responsiveness	NN	O	O
.	NN	O	O

Gel	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
detected	NN	O	O
an	NN	O	O
induced	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
by	NN	O	O
UV	NN	O	O
cross-linking	NN	O	O
,	NN	O	O
Southwestern	NN	O	O
blot	NN	O	O
,	NN	O	O
and	NN	O	O
antibody	NN	O	O
ablation/supershift	NN	O	O
assays	NN	O	O
revealed	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
a	NN	O	O
recently	NN	O	O
reported	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
designated	NN	O	O
SAF	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
LPS-mediated	NN	O	O
SAA	NN	O	B-protein
gene	NN	O	O
induction	NN	O	O
in	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
primarily	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
SAF	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cytomegalovirus	NN	O	B-DNA
immediate	NN	O	I-DNA
early	NN	O	I-DNA
genes	NN	O	I-DNA
upregulate	NN	O	O
interleukin-6	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
The	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
genes	NN	O	I-DNA
(	NN	O	O
IE	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
human	NN	O	O
cytomegalovirus	NN	O	O
(	NN	O	O
CMV	NN	O	O
)	NN	O	O
can	NN	O	O
be	NN	O	O
expressed	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
regulate	NN	O	O
viral	NN	O	B-DNA
and	NN	O	I-DNA
cellular	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Interleukin-6	NN	O	B-protein
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
plays	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
numerous	NN	O	O
inflammatory	NN	O	O
and	NN	O	O
immune	NN	O	O
processes	NN	O	O
.	NN	O	O

Interleukin-6	NN	O	B-protein
levels	NN	O	O
are	NN	O	O
increased	NN	O	O
in	NN	O	O
lung	NN	O	O
transplant	NN	O	O
patients	NN	O	O
clinically	NN	O	O
diagnosed	NN	O	O
with	NN	O	O
CMV	NN	O	O
pneumonitis	NN	O	O
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
various	NN	O	O
stimuli	NN	O	O
that	NN	O	O
include	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
viruses	NN	O	O
,	NN	O	O
and	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

These	NN	O	O
studies	NN	O	O
examined	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
CMV	NN	O	O
IE	NN	O	O
gene	NN	O	O
products	NN	O	O
to	NN	O	O
modulate	NN	O	O
IL-6	NN	O	B-protein
production	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
were	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
CMV	NN	O	B-DNA
IE	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Interleukin-6	NN	O	B-protein
protein	NN	O	O
and	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
were	NN	O	O
measured	NN	O	O
in	NN	O	O
control	NN	O	O
and	NN	O	O
CMV	NN	O	B-cell_type
immediate	NN	O	I-cell_type
early	NN	O	I-cell_type
transfected	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
CMV	NN	O	B-DNA
IE	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
IL-6	NN	O	B-DNA
chloramphenicol	NN	O	I-DNA
acetyl	NN	O	I-DNA
transferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
or	NN	O	I-DNA
IL-6	NN	O	I-DNA
luciferase	NN	O	I-DNA
constructs	NN	O	I-DNA
were	NN	O	O
used	NN	O	O
to	NN	O	O
study	NN	O	O
IL-6	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Interleukin-6	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
mRNA	NN	O	O
production	NN	O	O
were	NN	O	O
significantly	NN	O	O
increased	NN	O	O
in	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
CMV	NN	O	B-DNA
IE	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
stimulated	NN	O	O
with	NN	O	O
LPS	NN	O	O
compared	NN	O	O
to	NN	O	O
LPS-stimulated	NN	O	B-cell_line
control	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Cytomegalovirus	NN	O	B-protein
IE	NN	O	I-protein
gene	NN	O	I-protein
products	NN	O	I-protein
significantly	NN	O	O
enhanced	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
of	NN	O	O
IL-6	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
both	NN	O	O
IL-6	NN	O	B-protein
CAT	NN	O	O
and	NN	O	O
IL-6	NN	O	O
luciferase	NN	O	O
assays	NN	O	O
.	NN	O	O

A	NN	O	O
deletion	NN	O	O
construct	NN	O	O
that	NN	O	O
contains	NN	O	O
a	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
but	NN	O	O
is	NN	O	O
missing	NN	O	O
the	NN	O	O
multiple	NN	O	O
response	NN	O	O
region	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
continued	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-6	NN	O	B-protein
luciferase	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
LPS-stimulated	NN	O	B-cell_line
CMV	NN	O	I-cell_line
transfected	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Cytomegalovirus	NN	O	B-protein
immediate	NN	O	I-protein
early	NN	O	I-protein
gene	NN	O	I-protein
products	NN	O	I-protein
significantly	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
in	NN	O	O
LPS-stimulated	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-6	NN	O	B-protein
luciferase	NN	O	B-protein
activity	NN	O	O
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
multiple	NN	O	O
response	NN	O	O
region	NN	O	O
,	NN	O	O
the	NN	O	O
area	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
promoter	NN	O	I-DNA
responsive	NN	O	O
to	NN	O	O
IL-1	NN	O	B-protein
,	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
cyclic	NN	O	O
amp	NN	O	O
,	NN	O	O
and	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
CMV	NN	O	B-protein
IE	NN	O	I-protein
gene	NN	O	I-protein
products	NN	O	I-protein
to	NN	O	O
enhance	NN	O	O
IL-6	NN	O	B-protein
production	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
immune	NN	O	O
inflammatory	NN	O	O
states	NN	O	O
associated	NN	O	O
with	NN	O	O
CMV	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
relA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
subunit	NN	O	I-protein
expression	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
to	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
evaluated	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
subunits	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
(	NN	O	O
MDMs	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
consisting	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

An	NN	O	O
additional	NN	O	O
complex	NN	O	O
composed	NN	O	O
of	NN	O	O
p50/RelA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
heterodimers	NN	O	I-protein
appeared	NN	O	O
only	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
7-day	NN	O	O
MDMs	NN	O	B-cell_type
.	NN	O	O

Immunoblot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
p50	NN	O	O
subunit	NN	O	O
was	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
MDMs	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
RelA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
was	NN	O	O
barely	NN	O	O
detectable	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
its	NN	O	O
level	NN	O	O
increased	NN	O	O
markedly	NN	O	O
in	NN	O	O
MDMs	NN	O	B-cell_type
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
RelA	NN	O	B-RNA
(	NN	O	I-RNA
p65	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
RelA	NN	O	B-RNA
(	NN	O	I-RNA
p65	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
in	NN	O	O
MDMs	NN	O	B-cell_type
,	NN	O	O
compared	NN	O	O
with	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
MDMs	NN	O	B-cell_type
,	NN	O	O
an	NN	O	O
upregulation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
synthesis	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
M	NN	O	O
(	NN	O	O
r	NN	O	O
)	NN	O	O
40	NN	O	O
,	NN	O	O
000	NN	O	O
form	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
were	NN	O	O
also	NN	O	O
observed	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
active	NN	O	O
p50/RelA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
heterodimers	NN	O	I-protein
with	NN	O	O
the	NN	O	O
capacity	NN	O	O
to	NN	O	O
activate	NN	O	O
target	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
parallel	NN	O	O
induction	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
synthesis	NN	O	O
may	NN	O	O
allow	NN	O	O
for	NN	O	O
the	NN	O	O
continuous	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	O
reservoir	NN	O	O
of	NN	O	O
p50/RelA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
complexes	NN	O	I-protein
that	NN	O	O
are	NN	O	O
readily	NN	O	O
available	NN	O	O
for	NN	O	O
inducer-mediated	NN	O	O
stimulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
Rel-A	NN	O	I-protein
and	NN	O	I-protein
c-Rel	NN	O	I-protein
protein	NN	O	I-protein
complexes	NN	O	I-protein
are	NN	O	O
differentially	NN	O	O
distributed	NN	O	O
within	NN	O	O
human	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

Nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
/Rel	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
inducible	NN	O	O
transcriptional	NN	O	B-protein
regulators	NN	O	I-protein
of	NN	O	O
numerous	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

They	NN	O	O
are	NN	O	O
particularly	NN	O	O
abundant	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
and	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
reported	NN	O	O
previously	NN	O	O
that	NN	O	O
a	NN	O	O
nuclear	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
freshly	NN	O	O
extracted	NN	O	O
human	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
in	NN	O	O
vitro	NN	O	O
treatment	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
extracted	NN	O	O
from	NN	O	O
human	NN	O	O
thymocyte	NN	O	O
nuclei	NN	O	O
as	NN	O	O
being	NN	O	O
p50/p65	NN	O	B-protein
and	NN	O	I-protein
p50/c-Rel	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Immunochemical	NN	O	O
and	NN	O	O
immunofluorescent	NN	O	O
staining	NN	O	O
of	NN	O	O
thymus	NN	O	O
sections	NN	O	O
using	NN	O	O
specific	NN	O	B-protein
Abs	NN	O	I-protein
allowed	NN	O	O
visualization	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
both	NN	O	O
thymocytes	NN	O	B-cell_type
and	NN	O	O
nonthymocyte	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
detection	NN	O	O
suggested	NN	O	O
a	NN	O	O
preferential	NN	O	O
activation	NN	O	O
of	NN	O	O
p50/c-Rel	NN	O	B-protein
in	NN	O	O
medullary	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
p50/p65	NN	O	B-protein
was	NN	O	O
present	NN	O	O
in	NN	O	O
both	NN	O	O
cortical	NN	O	O
and	NN	O	O
medullary	NN	O	O
regions	NN	O	O
of	NN	O	O
human	NN	O	O
thymus	NN	O	O
lobules	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
intensity	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
labeling	NN	O	O
was	NN	O	O
much	NN	O	O
higher	NN	O	O
in	NN	O	O
several	NN	O	O
thymocytes	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
medulla	NN	O	O
.	NN	O	O

p65	NN	O	O
,	NN	O	O
p50	NN	O	O
,	NN	O	O
and	NN	O	O
c-Rel	NN	O	O
activities	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
both	NN	O	O
CD4-	NN	O	B-cell_type
and	NN	O	I-cell_type
CD8-positive	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
p65	NN	O	B-protein
and	NN	O	I-protein
c-Rel	NN	O	I-protein
complexes	NN	O	I-protein
play	NN	O	O
distinct	NN	O	O
roles	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
that	NN	O	O
both	NN	O	O
forms	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
play	NN	O	O
critical	NN	O	O
roles	NN	O	O
during	NN	O	O
late	NN	O	O
stages	NN	O	O
of	NN	O	O
the	NN	O	O
intrathymic	NN	O	O
maturation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

ALY	NN	O	B-protein
,	NN	O	O
a	NN	O	O
context-dependent	NN	O	B-protein
coactivator	NN	O	I-protein
of	NN	O	O
LEF-1	NN	O	B-protein
and	NN	O	O
AML-1	NN	O	B-protein
,	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
TCRalpha	NN	O	B-DNA
enhancer	NN	O	I-DNA
function	NN	O	O
.	NN	O	O

LEF-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
transcription	NN	O	O
factor	NN	O	O
that	NN	O	O
participates	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
TCR	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
by	NN	O	O
facilitating	NN	O	O
the	NN	O	O
assembly	NN	O	O
of	NN	O	O
multiple	NN	O	B-protein
proteins	NN	O	I-protein
into	NN	O	O
a	NN	O	O
higher	NN	O	O
order	NN	O	O
nucleoprotein	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

The	NN	O	O
function	NN	O	O
of	NN	O	O
LEF-1	NN	O	B-protein
is	NN	O	O
dependent	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
HMG	NN	O	B-protein
domain	NN	O	I-protein
that	NN	O	O
induces	NN	O	O
a	NN	O	O
sharp	NN	O	O
bend	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
helix	NN	O	O
,	NN	O	O
and	NN	O	O
on	NN	O	O
an	NN	O	O
activation	NN	O	B-protein
domain	NN	O	I-protein
that	NN	O	O
stimulates	NN	O	O
transcription	NN	O	O
only	NN	O	O
in	NN	O	O
a	NN	O	O
specific	NN	O	O
context	NN	O	O
of	NN	O	O
other	NN	O	O
enhancer-binding	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

With	NN	O	O
the	NN	O	O
aim	NN	O	O
of	NN	O	O
gaining	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
context-dependent	NN	O	B-protein
activation	NN	O	I-protein
domains	NN	O	I-protein
,	NN	O	O
we	NN	O	O
cloned	NN	O	O
ALY	NN	O	B-protein
,	NN	O	O
a	NN	O	O
novel	NN	O	O
LEF-1-interacting	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

ALY	NN	O	B-protein
is	NN	O	O
a	NN	O	O
ubiquitously	NN	O	O
expressed	NN	O	O
,	NN	O	O
nuclear	NN	O	O
protein	NN	O	O
that	NN	O	O
specifically	NN	O	O
associates	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
LEF-1	NN	O	B-protein
and	NN	O	O
AML-1	NN	O	B-protein
(	NN	O	O
CBF	NN	O	B-protein
alpha2	NN	O	I-protein
,	NN	O	O
PEBP2	NN	O	B-protein
alpha	NN	O	I-protein
(	NN	O	O
B	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
another	NN	O	O
protein	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
alpha	NN	O	I-protein
enhancer	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
ALY	NN	O	B-protein
can	NN	O	O
increase	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
both	NN	O	O
LEF-1	NN	O	B-protein
and	NN	O	O
AML	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
ALY	NN	O	B-protein
stimulates	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
alpha	NN	O	I-protein
enhancer	NN	O	I-protein
complex	NN	O	I-protein
reconstituted	NN	O	O
in	NN	O	O
transfected	NN	O	B-cell_line
nonlymphoid	NN	O	I-cell_line
HeLa	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
ALY	NN	O	B-protein
by	NN	O	O
anti-sense	NN	O	O
oligonucleotides	NN	O	O
virtually	NN	O	O
eliminates	NN	O	O
TCR	NN	O	B-DNA
alpha	NN	O	I-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
LEF-1	NN	O	B-protein
,	NN	O	O
ALY	NN	O	B-protein
can	NN	O	O
stimulate	NN	O	O
transcription	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
alpha	NN	O	I-DNA
enhancer	NN	O	I-DNA
but	NN	O	O
apparently	NN	O	O
not	NN	O	O
when	NN	O	O
tethered	NN	O	O
to	NN	O	O
DNA	NN	O	O
through	NN	O	O
an	NN	O	O
heterologous	NN	O	B-protein
DNA-binding	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
ALY	NN	O	B-protein
mediates	NN	O	O
context-dependent	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
facilitating	NN	O	O
the	NN	O	O
functional	NN	O	O
collaboration	NN	O	O
of	NN	O	O
multiple	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
alpha	NN	O	I-protein
enhancer	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

AML1a	NN	O	B-protein
but	NN	O	O
not	NN	O	O
AML1b	NN	O	B-protein
inhibits	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
sodium	NN	O	O
butyrate	NN	O	O
and	NN	O	O
enhances	NN	O	O
the	NN	O	O
megakaryocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

AML1	NN	O	B-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
cells	NN	O	O
along	NN	O	O
erythroid	NN	O	B-cell_type
and/or	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
,	NN	O	O
because	NN	O	O
a	NN	O	O
significant	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
AML1	NN	O	B-protein
gene	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

We	NN	O	O
overexpressed	NN	O	O
AML1a	NN	O	B-protein
(	NN	O	O
without	NN	O	O
the	NN	O	O
transcription-activating	NN	O	B-protein
domain	NN	O	I-protein
)	NN	O	O
and	NN	O	O
AML1b	NN	O	B-protein
(	NN	O	O
with	NN	O	O
the	NN	O	O
domain	NN	O	O
)	NN	O	O
proteins	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
into	NN	O	O
hemoglobin-producing	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
megakaryocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
AML1a-transfected	NN	O	B-cell_line
K562	NN	O	I-cell_line
cells	NN	O	I-cell_line
had	NN	O	O
a	NN	O	O
reduced	NN	O	O
capacity	NN	O	O
to	NN	O	O
differentiate	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
sodium	NN	O	O
n-butyrate	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
other	NN	O	O
inducers	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
hemin	NN	O	O
,	NN	O	O
1-beta-D-arabinofuranosylcytosine	NN	O	O
,	NN	O	O
and	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
.	NN	O	O

The	NN	O	O
AML1	NN	O	B-protein
antisense	NN	O	O
oligodeoxynucleotide	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
sense	NN	O	O
oligomer	NN	O	O
recovered	NN	O	O
its	NN	O	O
differentiation-inducing	NN	O	O
capacity	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
butyrate	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
AML1b	NN	O	B-protein
conferred	NN	O	O
a	NN	O	O
similar	NN	O	O
differentiation-inducing	NN	O	O
capacity	NN	O	O
upon	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
transfected	NN	O	O
with	NN	O	O
vector	NN	O	O
alone	NN	O	O
.	NN	O	O

AML1a	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
enhanced	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
megakaryocytic	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
AML1	NN	O	B-protein
proteins	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
erythroid	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
sequence	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
estrogen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
an	NN	O	O
ER-negative	NN	O	B-cell_line
breast	NN	O	I-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Given	NN	O	O
the	NN	O	O
important	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
and	NN	O	O
physiology	NN	O	O
of	NN	O	O
the	NN	O	O
breast	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
essential	NN	O	O
to	NN	O	O
delineate	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
responsible	NN	O	O
for	NN	O	O
its	NN	O	O
failed	NN	O	O
expression	NN	O	O
in	NN	O	O
some	NN	O	O
breast	NN	O	O
tumors	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
a	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
ER	NN	O	B-protein
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
ER-positive	NN	O	B-cell_line
MCF-7	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
ER-negative	NN	O	B-cell_line
MDA-MB-231	NN	O	I-cell_line
breast	NN	O	I-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
to	NN	O	O
determine	NN	O	O
if	NN	O	O
sequence	NN	O	O
alterations	NN	O	O
in	NN	O	O
this	NN	O	O
region	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
ER	NN	O	B-protein
-negative	NN	O	O
phenotype	NN	O	O
of	NN	O	O
some	NN	O	O
tumors	NN	O	O
.	NN	O	O

From	NN	O	O
this	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
variations	NN	O	O
between	NN	O	O
the	NN	O	O
sequences	NN	O	O
,	NN	O	O
two	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
determined	NN	O	O
to	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
50	NN	O	O
%	NN	O	O
decrease	NN	O	O
in	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
by	NN	O	O
azelastine	NN	O	O
hydrochloride	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
involved	NN	O	O
in	NN	O	O
generation	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
.	NN	O	O

The	NN	O	O
influence	NN	O	O
of	NN	O	O
the	NN	O	O
anti-allergy	NN	O	O
agent	NN	O	O
azelastine	NN	O	O
hydrochloride	NN	O	O
(	NN	O	O
Azeptin	NN	O	O
)	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
(	NN	O	O
NO	NN	O	O
)	NN	O	O
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
various	NN	O	O
kinds	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
and	NN	O	I-cell_type
mouse	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Azeptin	NN	O	O
dose-dependently	NN	O	O
suppressed	NN	O	O
both	NN	O	O
DNA	NN	O	O
and	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
gingival	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
(	NN	O	O
HF	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
also	NN	O	O
suppressed	NN	O	O
blastogenesis	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
PBL	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O
Generation	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
,	NN	O	O
interleukin	NN	O	B-protein
1-beta	NN	O	I-protein
,	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
interleukin-6	NN	O	B-protein
from	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
M	NN	O	O
Azeptin-treated	NN	O	O
PBL	NN	O	B-cell_type
and	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
(	NN	O	O
HM	NN	O	B-cell_type
)	NN	O	O
was	NN	O	O
decreased	NN	O	O
to	NN	O	O
approximately	NN	O	O
1/3	NN	O	O
to	NN	O	O
2/3	NN	O	O
of	NN	O	O
the	NN	O	O
control	NN	O	O
levels	NN	O	O
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
with	NN	O	O
the	NN	O	O
decreased	NN	O	O
cytokine	NN	O	B-protein
generation	NN	O	O
,	NN	O	O
each	NN	O	O
cytokine	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
less	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
M	NN	O	O
Azeptin	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
both	NN	O	O
inducible	NN	O	O
nitric	NN	O	B-protein
oxide	NN	O	I-protein
synthase	NN	O	I-protein
-mRNA	NN	O	O
level	NN	O	O
and	NN	O	O
NO	NN	O	O
generation	NN	O	O
in	NN	O	O
mouse	NN	O	B-cell_type
peritoneal	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
were	NN	O	O
suppressed	NN	O	O
by	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
M	NN	O	O
Azeptin	NN	O	O
.	NN	O	O

Being	NN	O	O
compatible	NN	O	O
with	NN	O	O
those	NN	O	O
results	NN	O	O
,	NN	O	O
Azeptin	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
suppressed	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
PBL	NN	O	B-cell_type
,	NN	O	O
HM	NN	O	O
and	NN	O	O
HF	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
appear	NN	O	O
to	NN	O	O
indicate	NN	O	O
that	NN	O	O
suppression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
and	NN	O	O
NO	NN	O	O
generation	NN	O	O
by	NN	O	O
Azeptin	NN	O	O
results	NN	O	O
at	NN	O	O
least	NN	O	O
partially	NN	O	O
from	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
interaction	NN	O	O
of	NN	O	O
nuclear	NN	O	O
factors	NN	O	O
with	NN	O	O
the	NN	O	O
PRE-I	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
different	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
subsets	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
immunomodulatory	NN	O	O
cytokine	NN	O	B-protein
IL-4	NN	O	B-protein
affects	NN	O	O
cells	NN	O	O
of	NN	O	O
most	NN	O	O
hemopoietic	NN	O	B-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

IL-4	NN	O	B-protein
is	NN	O	O
secreted	NN	O	O
by	NN	O	O
activated	NN	O	O
Th2	NN	O	B-cell_type
but	NN	O	O
not	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
plays	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
by	NN	O	O
modulating	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
Th	NN	O	I-cell_type
cells	NN	O	I-cell_type
toward	NN	O	O
the	NN	O	O
Th2	NN	O	B-cell_type
phenotype	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
identified	NN	O	O
an	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
PRE-I	NN	O	B-DNA
,	NN	O	O
that	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
responsible	NN	O	O
for	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
gene	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
PRE-I	NN	O	B-DNA
site	NN	O	I-DNA
and	NN	O	O
compared	NN	O	O
the	NN	O	O
binding	NN	O	O
activities	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
to	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
promoter	NN	O	O
of	NN	O	O
Th1	NN	O	B-cell_type
and	NN	O	I-cell_type
Th2	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
PRE-I	NN	O	B-DNA
interacts	NN	O	O
with	NN	O	O
PMA	NN	O	O
-and	NN	O	O
PMA/ionomycin-inducible	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A-sensitive	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
anti-C/EBPbeta	NN	O	B-protein
(	NN	O	O
NF-IL6	NN	O	B-protein
)	NN	O	O
,	NN	O	O
anti-C/EBPdelta	NN	O	B-protein
(	NN	O	O
NF-IL6beta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
anti-NF-ATc	NN	O	B-protein
,	NN	O	O
anti-NF-ATp	NN	O	B-protein
,	NN	O	O
anti-Fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
anti-Jun	NN	O	B-protein
Abs	NN	O	I-protein
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
previously	NN	O	O
identified	NN	O	O
PRE-I	NN	O	B-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
POS-1	NN	O	B-protein
is	NN	O	O
composed	NN	O	O
of	NN	O	O
different	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
different	NN	O	O
Th	NN	O	O
cell	NN	O	O
subsets	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
IL-4-producing	NN	O	B-cell_line
Th0-like	NN	O	I-cell_line
human	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
and	NN	O	O
mouse	NN	O	B-cell_line
EL-4	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
POS-1	NN	O	B-protein
(	NN	O	O
designated	NN	O	O
POS-1a	NN	O	B-protein
)	NN	O	O
contains	NN	O	O
NF-IL6beta	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
mouse	NN	O	B-cell_line
Th2	NN	O	I-cell_line
D10	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
Th2	NN	O	I-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
POS-1	NN	O	B-protein
(	NN	O	O
designated	NN	O	O
POS-1b	NN	O	B-protein
)	NN	O	O
contains	NN	O	O
NF-IL6beta	NN	O	B-protein
,	NN	O	O
Jun	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-ATc/p	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
POS-1	NN	O	B-protein
was	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
Th1	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
PRE-I	NN	O	B-DNA
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
differential	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
levels	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
human	NN	O	O
epsilon	NN	O	O
germline	NN	O	B-DNA
transcription	NN	O	O
:	NN	O	O
role	NN	O	O
of	NN	O	O
B-cell-specific	NN	O	B-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Germline	NN	O	B-protein
transcripts	NN	O	I-protein
initiate	NN	O	O
from	NN	O	O
promoters	NN	O	B-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
switch	NN	O	I-DNA
region	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
are	NN	O	O
necessary	NN	O	O
to	NN	O	O
target	NN	O	O
the	NN	O	O
appropriate	NN	O	O
switch	NN	O	O
region	NN	O	O
for	NN	O	O
recombination	NN	O	O
and	NN	O	O
switching	NN	O	O
.	NN	O	O

Different	NN	O	O
cytokines	NN	O	B-protein
activate	NN	O	O
transcription	NN	O	O
at	NN	O	O
the	NN	O	O
appropriate	NN	O	O
germline	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Because	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
B-cell-specific	NN	O	B-protein
activator	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
BSAP	NN	O	B-protein
)	NN	O	O
are	NN	O	O
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
several	NN	O	O
switch	NN	O	O
regions	NN	O	O
in	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
cluster	NN	O	I-DNA
,	NN	O	O
BSAP	NN	O	B-protein
might	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
germline	NN	O	B-DNA
transcription	NN	O	O
and	NN	O	O
isotype	NN	O	O
switching	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
whether	NN	O	O
BSAP	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
epsilon	NN	O	O
germline	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
BSAP	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
both	NN	O	O
IL-4	NN	O	B-protein
-dependent	NN	O	O
induction	NN	O	O
and	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
upregulation	NN	O	O
of	NN	O	O
human	NN	O	O
epsilon	NN	O	O
germline	NN	O	B-DNA
transcription	NN	O	O
.	NN	O	O

BSAP	NN	O	B-protein
is	NN	O	O
unique	NN	O	O
among	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
epsilon	NN	O	O
germline	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
because	NN	O	O
it	NN	O	O
is	NN	O	O
B	NN	O	O
cell	NN	O	O
specific	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
at	NN	O	O
the	NN	O	O
merging	NN	O	O
point	NN	O	O
of	NN	O	O
two	NN	O	O
signalling	NN	O	O
pathways	NN	O	O
that	NN	O	O
are	NN	O	O
critical	NN	O	O
for	NN	O	O
IgE	NN	O	B-protein
switching	NN	O	O
.	NN	O	O

-DOCSTART-	O

Experimental	NN	O	O
expression	NN	O	O
in	NN	O	O
mice	NN	O	O
and	NN	O	O
spontaneous	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	O
SLE	NN	O	O
of	NN	O	O
polyomavirus	NN	O	B-protein
T-antigen	NN	O	I-protein
.	NN	O	O

A	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	O
DNA	NN	O	O
and	NN	O	O
eukaryotic	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
experimental	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
polyomavirus	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
T-antigen	NN	O	I-protein
has	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
induce	NN	O	O
anti-DNA	NN	O	B-protein
antibodies	NN	O	I-protein
in	NN	O	O
mice	NN	O	O
.	NN	O	O

Two	NN	O	O
sets	NN	O	O
of	NN	O	O
independent	NN	O	O
evidences	NN	O	O
are	NN	O	O
presented	NN	O	O
here	NN	O	O
that	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
biological	NN	O	O
relevance	NN	O	O
for	NN	O	O
this	NN	O	O
model	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
results	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
mice	NN	O	O
inoculated	NN	O	O
with	NN	O	O
T-antigen-expressing	NN	O	B-DNA
plasmids	NN	O	I-DNA
produced	NN	O	O
antibodies	NN	O	B-protein
,	NN	O	O
not	NN	O	O
only	NN	O	O
to	NN	O	O
T-antigen	NN	O	B-protein
and	NN	O	O
DNA	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
to	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
eukaryotic	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
TATA-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
TBP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
the	NN	O	O
cAMP-response-element-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Secondly	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
polyomavirus	NN	O	O
reactivation	NN	O	O
occurs	NN	O	O
in	NN	O	O
SLE	NN	O	O
patients	NN	O	O
,	NN	O	O
and	NN	O	O
whether	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	O
T-antigen	NN	O	B-protein
,	NN	O	O
DNA	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
TBP	NN	O	B-protein
and	NN	O	O
CREB	NN	O	B-protein
are	NN	O	O
linked	NN	O	O
to	NN	O	O
such	NN	O	O
events	NN	O	O
.	NN	O	O

Both	NN	O	O
within	NN	O	O
and	NN	O	O
among	NN	O	O
these	NN	O	O
SLE	NN	O	O
patients	NN	O	O
,	NN	O	O
frequent	NN	O	O
polyomavirus	NN	O	O
reactivations	NN	O	O
were	NN	O	O
observed	NN	O	O
that	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
certain	NN	O	O
rearrangements	NN	O	O
of	NN	O	O
the	NN	O	O
noncoding	NN	O	O
control	NN	O	O
regions	NN	O	O
,	NN	O	O
nor	NN	O	O
by	NN	O	O
corticosteroid	NN	O	O
treatment	NN	O	O
.	NN	O	O

Linked	NN	O	O
to	NN	O	O
these	NN	O	O
events	NN	O	O
,	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	O
T-antigen	NN	O	B-protein
,	NN	O	O
DNA	NN	O	O
,	NN	O	O
TBP	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CREB	NN	O	B-protein
were	NN	O	O
detected	NN	O	O
,	NN	O	O
identical	NN	O	O
to	NN	O	O
what	NN	O	O
we	NN	O	O
observed	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

Antibodies	NN	O	O
recognizing	NN	O	O
double-stranded	NN	O	O
DNA	NN	O	O
were	NN	O	O
confined	NN	O	O
to	NN	O	O
patients	NN	O	O
with	NN	O	O
frequent	NN	O	O
polyomavirus	NN	O	O
reactivations	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
described	NN	O	O
here	NN	O	O
indicate	NN	O	O
that	NN	O	O
cognate	NN	O	O
interaction	NN	O	O
of	NN	O	O
B	NN	O	O
cells	NN	O	O
recognizing	NN	O	O
DNA	NN	O	O
or	NN	O	O
DNA-associated	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
recognizing	NN	O	O
T	NN	O	B-protein
antigen	NN	O	I-protein
had	NN	O	O
taken	NN	O	O
place	NN	O	O
as	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
complex	NN	O	O
formation	NN	O	O
between	NN	O	O
T	NN	O	B-protein
ag	NN	O	I-protein
and	NN	O	O
DNA	NN	O	O
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
polyomavirus	NN	O	O
reactivations	NN	O	O
.	NN	O	O

-DOCSTART-	O

Jak3	NN	O	B-protein
is	NN	O	O
associated	NN	O	O
with	NN	O	O
CD40	NN	O	B-protein
and	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
CD40	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CD40	NN	O	B-protein
is	NN	O	O
a	NN	O	O
receptor	NN	O	O
that	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
survival	NN	O	O
,	NN	O	O
growth	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
isotype	NN	O	O
switching	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
CD40	NN	O	B-protein
lacks	NN	O	O
intrinsic	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
activity	NN	O	O
,	NN	O	O
its	NN	O	O
ligation	NN	O	O
induces	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
several	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
events	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
engagement	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
Jak3	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
of	NN	O	O
STAT3	NN	O	B-protein
.	NN	O	O

Jak3	NN	O	B-protein
is	NN	O	O
constitutively	NN	O	O
associated	NN	O	O
with	NN	O	O
CD40	NN	O	B-protein
,	NN	O	O
and	NN	O	O
this	NN	O	O
interaction	NN	O	O
requires	NN	O	O
a	NN	O	O
proline-rich	NN	O	B-protein
sequence	NN	O	I-protein
in	NN	O	O
the	NN	O	O
membrane-proximal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
CD40	NN	O	B-protein
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
this	NN	O	O
sequence	NN	O	O
abolishes	NN	O	O
the	NN	O	O
capacity	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
CD23	NN	O	B-protein
,	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
lymphotoxin-alpha	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
signaling	NN	O	O
through	NN	O	O
Jak3	NN	O	B-protein
is	NN	O	O
activated	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
and	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
functions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
required	NN	O	O
for	NN	O	O
lymphoid	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
.	NN	O	O

Intimate	NN	O	O
interactions	NN	O	O
between	NN	O	O
multipotential	NN	O	O
hemopoietic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
their	NN	O	O
microenvironment	NN	O	O
work	NN	O	O
towards	NN	O	O
redefining	NN	O	O
the	NN	O	O
identity	NN	O	O
and	NN	O	O
the	NN	O	O
differentiative	NN	O	O
fate	NN	O	O
of	NN	O	O
these	NN	O	O
primitive	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Molecular	NN	O	O
cues	NN	O	O
delivered	NN	O	O
by	NN	O	O
the	NN	O	O
microenvironment	NN	O	O
frequently	NN	O	O
act	NN	O	O
in	NN	O	O
an	NN	O	O
instructive	NN	O	O
fashion	NN	O	O
by	NN	O	O
initiating	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
that	NN	O	O
ultimately	NN	O	O
target	NN	O	O
a	NN	O	O
select	NN	O	O
group	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

These	NN	O	O
transcriptional	NN	O	O
regulators	NN	O	O
in	NN	O	O
turn	NN	O	O
trigger	NN	O	O
a	NN	O	O
cascade	NN	O	O
of	NN	O	O
genetic	NN	O	O
changes	NN	O	O
that	NN	O	O
ultimately	NN	O	O
determine	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
during	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Gene	NN	O	O
inactivation	NN	O	O
studies	NN	O	O
on	NN	O	O
the	NN	O	O
PU.1	NN	O	B-DNA
,	NN	O	I-DNA
Ikaros	NN	O	I-DNA
and	NN	O	I-DNA
GATA-3	NN	O	I-DNA
genes	NN	O	I-DNA
have	NN	O	O
revealed	NN	O	O
that	NN	O	O
their	NN	O	O
encoded	NN	O	O
factors	NN	O	O
are	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
earliest	NN	O	O
commitment	NN	O	O
step	NN	O	O
into	NN	O	O
the	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Interleukin-4	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
X-linked	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
.	NN	O	O

Interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
cytokine	NN	O	O
for	NN	O	O
B	NN	O	O
and	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
function	NN	O	O
and	NN	O	O
mediates	NN	O	O
its	NN	O	O
effects	NN	O	O
via	NN	O	O
a	NN	O	O
receptor	NN	O	O
that	NN	O	O
contains	NN	O	O
gammac	NN	O	B-protein
.	NN	O	O

B	NN	O	B-cell_type
cells	NN	O	I-cell_type
derived	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
X-linked	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
(	NN	O	O
X-SCID	NN	O	O
)	NN	O	O
are	NN	O	O
deficient	NN	O	O
in	NN	O	O
gammac	NN	O	O
and	NN	O	O
provide	NN	O	O
a	NN	O	O
useful	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
to	NN	O	O
dissect	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
subunit	NN	O	O
in	NN	O	O
IL-4	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
although	NN	O	O
IL-4	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
X-SCID	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
did	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
Janus	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase-3	NN	O	I-protein
(	NN	O	O
JAK3	NN	O	B-protein
)	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
other	NN	O	O
IL-4	NN	O	B-protein
substrates	NN	O	I-protein
including	NN	O	O
JAK1	NN	O	B-protein
and	NN	O	O
IRS-1	NN	O	B-protein
were	NN	O	O
phosphorylated	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
we	NN	O	O
detected	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
6	NN	O	I-protein
(	NN	O	O
STAT6	NN	O	B-protein
)	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
X-SCID	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
gammac	NN	O	B-protein
mutations	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
reconstitution	NN	O	O
of	NN	O	O
these	NN	O	O
X-SCID	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
gammac	NN	O	B-protein
enhanced	NN	O	O
IL-4	NN	O	B-protein
-mediated	NN	O	O
responses	NN	O	O
including	NN	O	O
STAT6	NN	O	B-protein
phosphorylation	NN	O	O
and	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
resulted	NN	O	O
in	NN	O	O
increased	NN	O	O
CD23	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
gammac	NN	O	B-protein
is	NN	O	O
not	NN	O	O
necessary	NN	O	O
to	NN	O	O
trigger	NN	O	O
IL-4	NN	O	B-protein
-mediated	NN	O	O
responses	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
its	NN	O	O
presence	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
optimal	NN	O	O
IL-4	NN	O	B-protein
-signaling	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
two	NN	O	O
distinct	NN	O	O
IL-4	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
exist	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cell-to-cell	NN	O	O
contact	NN	O	O
activates	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
1	NN	O	O
through	NN	O	O
its	NN	O	O
kappaB	NN	O	B-DNA
motif	NN	O	I-DNA
.	NN	O	O

Cell-to-cell	NN	O	O
contact	NN	O	O
between	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
transfected	NN	O	B-cell_line
human	NN	O	I-cell_line
colonic	NN	O	I-cell_line
carcinoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
HT29	NN	O	I-cell_line
activates	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeats	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
.	NN	O	O

HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
transcription	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
complex	NN	O	O
array	NN	O	O
of	NN	O	O
virus-encoded	NN	O	B-protein
and	NN	O	I-protein
cellular	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
various	NN	O	O
constructs	NN	O	O
expressing	NN	O	O
a	NN	O	O
lacZ	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
intact	NN	O	O
or	NN	O	O
three	NN	O	O
deleted	NN	O	O
forms	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
obtained	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
kappaB	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
located	NN	O	O
in	NN	O	O
the	NN	O	O
U3	NN	O	B-DNA
region	NN	O	I-DNA
are	NN	O	O
involved	NN	O	O
in	NN	O	O
cell-to-cell	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

Cell-to-cell	NN	O	O
contact	NN	O	O
activates	NN	O	O
in	NN	O	O
vitro	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	I-protein
NF-kappaB	NN	O	I-protein
)	NN	O	I-protein
p50/p65	NN	O	I-protein
heterodimer	NN	O	I-protein
to	NN	O	O
an	NN	O	O
HIV-1	NN	O	O
kappaB	NN	O	O
oligonucleotide	NN	O	O
.	NN	O	O

Cell-to-cell	NN	O	O
contact	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
was	NN	O	O
only	NN	O	O
partially	NN	O	O
inhibited	NN	O	O
by	NN	O	O
100	NN	O	O
microM	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
and	NN	O	O
was	NN	O	O
not	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
inhibitor	NN	O	I-protein
kappaB	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

NF-kappaB	NN	O	B-protein
nuclear	NN	O	O
activation	NN	O	O
was	NN	O	O
not	NN	O	O
detectable	NN	O	O
before	NN	O	O
1	NN	O	O
h	NN	O	O
after	NN	O	O
cell	NN	O	O
contact	NN	O	O
and	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
nucleotide	NN	O	B-DNA
sequences	NN	O	I-DNA
that	NN	O	O
regulate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
MCF13	NN	O	O
murine	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
in	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
region	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
(	NN	O	O
DEN	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
mink	NN	O	O
cell	NN	O	O
focus-forming	NN	O	O
murine	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
viral	NN	O	O
pathogenicity	NN	O	O
.	NN	O	O

Another	NN	O	O
important	NN	O	O
activity	NN	O	O
of	NN	O	O
DEN	NN	O	B-DNA
is	NN	O	O
its	NN	O	O
control	NN	O	O
of	NN	O	O
transcription	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
we	NN	O	O
have	NN	O	O
determined	NN	O	O
that	NN	O	O
an	NN	O	O
NF-kappaB	NN	O	B-protein
site	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
this	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Possible	NN	O	O
role	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
germline	NN	O	B-DNA
C	NN	O	O
epsilon	NN	O	O
transcription	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
Burkitt	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
plays	NN	O	O
a	NN	O	O
broad	NN	O	O
role	NN	O	O
in	NN	O	O
gene	NN	O	O
regulation	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
evident	NN	O	O
whether	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
acts	NN	O	O
as	NN	O	O
a	NN	O	O
messenger	NN	O	O
system	NN	O	O
for	NN	O	O
germline	NN	O	B-DNA
C	NN	O	O
epsilon	NN	O	O
transcription	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
signaling	NN	O	O
cascade	NN	O	O
triggered	NN	O	O
by	NN	O	O
interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
or	NN	O	O
anti-CD40	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
mAb	NN	O	B-protein
)	NN	O	O
participates	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
responsible	NN	O	O
for	NN	O	O
germline	NN	O	B-DNA
C	NN	O	O
epsilon	NN	O	O
transcription	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
Burkitt	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
DND39	NN	O	B-cell_line
.	NN	O	O

Both	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
anti-	NN	O	O
CD40	NN	O	B-protein
mAb	NN	O	I-protein
induced	NN	O	O
activation	NN	O	O
of	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
(	NN	O	O
PI3-kinase	NN	O	B-protein
)	NN	O	O
,	NN	O	O
translocation	NN	O	O
of	NN	O	O
a	NN	O	O
zeta	NN	O	O
isoform	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
,	NN	O	O
and	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

All	NN	O	O
such	NN	O	O
events	NN	O	O
were	NN	O	O
abrogated	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
LY294002	NN	O	O
,	NN	O	O
a	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
PI3-kinase	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
N-acetyl-L-cysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
antioxidant	NN	O	O
,	NN	O	O
decreased	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
caused	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
anti-CD40	NN	O	B-protein
mAb	NN	O	I-protein
,	NN	O	O
or	NN	O	O
their	NN	O	O
combination	NN	O	O
.	NN	O	O

NAC	NN	O	O
was	NN	O	O
also	NN	O	O
effective	NN	O	O
in	NN	O	O
diminishing	NN	O	O
germline	NN	O	B-DNA
C	NN	O	O
epsilon	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
potency	NN	O	O
was	NN	O	O
higher	NN	O	O
in	NN	O	O
cultures	NN	O	O
costimulated	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
anti-CD40	NN	O	B-protein
mAb	NN	O	I-protein
than	NN	O	O
in	NN	O	O
those	NN	O	O
stimulated	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
ligation	NN	O	O
of	NN	O	O
CD40	NN	O	B-protein
induce	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
expression	NN	O	O
via	NN	O	O
at	NN	O	O
least	NN	O	O
a	NN	O	O
mechanism	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
PI3-kinase	NN	O	B-protein
pathway	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
sensitive	NN	O	O
to	NN	O	O
NAC	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
germline	NN	O	B-DNA
C	NN	O	O
epsilon	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differentiation	NN	O	O
of	NN	O	O
U-937	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
etoposide	NN	O	O
and	NN	O	O
ICRF-193	NN	O	O
,	NN	O	O
two	NN	O	O
antitumour	NN	O	O
DNA	NN	O	B-protein
topoisomerase	NN	O	I-protein
II	NN	O	I-protein
inhibitors	NN	O	O
with	NN	O	O
different	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
action	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
compared	NN	O	O
the	NN	O	O
action	NN	O	O
on	NN	O	O
U-937	NN	O	B-cell_line
human	NN	O	I-cell_line
promonocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
two	NN	O	O
DNA	NN	O	B-protein
topoisomerase	NN	O	I-protein
II	NN	O	I-protein
inhibitors	NN	O	O
,	NN	O	O
namely	NN	O	O
the	NN	O	O
epipodophyllotoxin	NN	O	O
etoposide	NN	O	O
and	NN	O	O
the	NN	O	O
bisdioxopiperazine	NN	O	O
ICRF-193	NN	O	O
.	NN	O	O

One	NN	O	O
hour	NN	O	O
pulse-treatment	NN	O	O
with	NN	O	O
3	NN	O	O
microM	NN	O	O
etoposide	NN	O	O
caused	NN	O	O
topoisomerase	NN	O	B-protein
associated	NN	O	O
,	NN	O	O
primary	NN	O	O
DNA	NN	O	O
breakage	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
rapidly	NN	O	O
followed	NN	O	O
by	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
these	NN	O	O
effects	NN	O	O
were	NN	O	O
not	NN	O	O
observed	NN	O	O
upon	NN	O	O
pulse-treatment	NN	O	O
with	NN	O	O
6	NN	O	O
microM	NN	O	O
ICRF-193	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
continuous	NN	O	O
treatments	NN	O	O
with	NN	O	O
subcytotoxic	NN	O	O
concentrations	NN	O	O
of	NN	O	O
etoposide	NN	O	O
(	NN	O	O
0.15	NN	O	O
microM	NN	O	O
)	NN	O	O
and	NN	O	O
ICRF-193	NN	O	O
(	NN	O	O
0.3	NN	O	O
microM	NN	O	O
)	NN	O	O
produced	NN	O	O
several	NN	O	O
similar	NN	O	O
effects	NN	O	O
,	NN	O	O
namely	NN	O	O
decreased	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
accumulation	NN	O	O
of	NN	O	O
cells	NN	O	O
at	NN	O	O
G2	NN	O	O
,	NN	O	O
increase	NN	O	O
in	NN	O	O
cell	NN	O	O
mass	NN	O	O
,	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Under	NN	O	O
these	NN	O	O
conditions	NN	O	O
,	NN	O	O
etoposide	NN	O	O
produced	NN	O	O
a	NN	O	O
biphasic	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
,	NN	O	O
which	NN	O	O
consisted	NN	O	O
in	NN	O	O
an	NN	O	O
early	NN	O	O
transient	NN	O	O
activation	NN	O	O
(	NN	O	O
from	NN	O	O
hours	NN	O	O
1	NN	O	O
to	NN	O	O
6	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
membrane-bound	NN	O	B-protein
enzyme	NN	O	I-protein
followed	NN	O	O
by	NN	O	O
a	NN	O	O
later	NN	O	O
activation	NN	O	O
(	NN	O	O
hour	NN	O	O
48	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
total	NN	O	O
,	NN	O	O
membrane-bound	NN	O	B-protein
and	NN	O	I-protein
cytosolic	NN	O	I-protein
enzyme	NN	O	I-protein
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
ICRF-193	NN	O	O
only	NN	O	O
provoked	NN	O	O
a	NN	O	O
late	NN	O	O
activation	NN	O	O
(	NN	O	O
from	NN	O	O
hours	NN	O	O
72	NN	O	O
to	NN	O	O
96	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
total	NN	O	O
enzyme	NN	O	O
.	NN	O	O

When	NN	O	O
used	NN	O	O
at	NN	O	O
differentiation-inducing	NN	O	O
concentrations	NN	O	O
,	NN	O	O
both	NN	O	O
topoisomerase	NN	O	B-protein
inhibitors	NN	O	O
caused	NN	O	O
a	NN	O	O
great	NN	O	O
stimulation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
with	NN	O	O
maximum	NN	O	O
value	NN	O	O
at	NN	O	O
hour	NN	O	O
12	NN	O	O
in	NN	O	O
etoposide-treated	NN	O	O
cells	NN	O	O
and	NN	O	O
at	NN	O	O
hour	NN	O	O
48	NN	O	O
in	NN	O	O
ICRF-193-treated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
(	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
EGR-1	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
was	NN	O	O
little	NN	O	O
affected	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
topoisomerase	NN	O	O
II	NN	O	O
inhibitors	NN	O	O
may	NN	O	O
induce	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
promonocytic	NN	O	O
cells	NN	O	O
,	NN	O	O
independently	NN	O	O
of	NN	O	O
their	NN	O	O
capacity	NN	O	O
to	NN	O	O
cause	NN	O	O
DNA	NN	O	O
strand	NN	O	O
breaks	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
there	NN	O	O
are	NN	O	O
other	NN	O	O
effects	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
early	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
,	NN	O	O
which	NN	O	O
are	NN	O	O
probably	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
primary	NN	O	O
DNA	NN	O	O
breakage	NN	O	O
by	NN	O	O
some	NN	O	O
anti-	NN	O	O
topoisomerase	NN	O	B-protein
drugs	NN	O	O
.	NN	O	O

-DOCSTART-	O

GATA-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
down-regulated	NN	O	O
in	NN	O	O
late	NN	O	O
S	NN	O	O
phase	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
set	NN	O	O
out	NN	O	O
to	NN	O	O
test	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
tissue-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
that	NN	O	O
relies	NN	O	O
on	NN	O	O
the	NN	O	O
early	NN	O	O
replication	NN	O	O
of	NN	O	O
expressed	NN	O	O
genes	NN	O	O
to	NN	O	O
sequester	NN	O	O
limiting	NN	O	O
activating	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
an	NN	O	O
erythroid	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
have	NN	O	O
tested	NN	O	O
the	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
lineage-restricted	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
through	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
GATA-1	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
low	NN	O	O
in	NN	O	O
G1	NN	O	O
,	NN	O	O
peaks	NN	O	O
in	NN	O	O
mid-S	NN	O	O
phase	NN	O	O
,	NN	O	O
and	NN	O	O
then	NN	O	O
decreases	NN	O	O
in	NN	O	O
G2/M	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
activities	NN	O	O
of	NN	O	O
two	NN	O	O
ubiquitous	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
Oct1	NN	O	B-protein
and	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
remain	NN	O	O
high	NN	O	O
in	NN	O	O
G2/M	NN	O	O
.	NN	O	O

GATA-1	NN	O	B-protein
protein	NN	O	O
and	NN	O	O
mRNA	NN	O	B-RNA
vary	NN	O	O
in	NN	O	O
a	NN	O	O
similar	NN	O	O
manner	NN	O	O
through	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
or	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
its	NN	O	O
message	NN	O	O
is	NN	O	O
regulated	NN	O	O
.	NN	O	O

Although	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
or	NN	O	O
DNA	NN	O	O
replication	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
peak	NN	O	O
in	NN	O	O
S	NN	O	O
phase	NN	O	O
,	NN	O	O
this	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
example	NN	O	O
of	NN	O	O
a	NN	O	O
lineage-restricted	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
displaying	NN	O	O
S	NN	O	O
phase-specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

One	NN	O	O
interpretation	NN	O	O
of	NN	O	O
these	NN	O	O
data	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
peak	NN	O	O
in	NN	O	O
GATA-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
amplifies	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
early	NN	O	O
replication	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
erythroid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
same	NN	O	O
time	NN	O	O
as	NN	O	O
preventing	NN	O	O
activation	NN	O	O
of	NN	O	O
non-erythroid	NN	O	B-DNA
genes	NN	O	I-DNA
containing	NN	O	O
GATA-responsive	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
may	NN	O	O
also	NN	O	O
relate	NN	O	O
to	NN	O	O
recent	NN	O	O
data	NN	O	O
implicating	NN	O	O
GATA-1	NN	O	B-protein
function	NN	O	O
in	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
pathway	NN	O	O
by	NN	O	O
inflammatory	NN	O	O
stimuli	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

Activated	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
have	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
upregulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
genes	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
those	NN	O	O
encoding	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
chemokines	NN	O	B-protein
,	NN	O	O
and	NN	O	O
to	NN	O	O
subsequently	NN	O	O
release	NN	O	O
the	NN	O	O
corresponding	NN	O	O
proteins	NN	O	O
.	NN	O	O

Although	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
date	NN	O	O
concerning	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
it	NN	O	O
is	NN	O	O
noteworthy	NN	O	O
that	NN	O	O
many	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
depend	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
for	NN	O	O
inducible	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
investigated	NN	O	O
whether	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
their	NN	O	O
fate	NN	O	O
on	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
that	NN	O	O
dimers	NN	O	O
consisting	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
NFkappaB1	NN	O	I-protein
,	NN	O	O
p65	NN	O	B-protein
RelA	NN	O	I-protein
,	NN	O	O
and/or	NN	O	O
c-Rel	NN	O	B-protein
are	NN	O	O
present	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
and	NN	O	O
that	NN	O	O
the	NN	O	O
greater	NN	O	O
part	NN	O	O
of	NN	O	O
these	NN	O	O
protein	NN	O	B-protein
complexes	NN	O	I-protein
is	NN	O	O
physically	NN	O	O
associated	NN	O	O
with	NN	O	O
cytoplasmic	NN	O	B-protein
IkappaB-alpha	NN	O	I-protein
in	NN	O	O
resting	NN	O	O
cells	NN	O	O
.	NN	O	O

Following	NN	O	O
neutrophil	NN	O	O
stimulation	NN	O	O
with	NN	O	O
proinflammatory	NN	O	O
agonists	NN	O	O
(	NN	O	O
such	NN	O	O
as	NN	O	O
lipopolysaccharide	NN	O	O
[	NN	O	O
LPS	NN	O	O
]	NN	O	O
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
[	NN	O	O
TNF-alpha	NN	O	B-protein
]	NN	O	O
,	NN	O	O
and	NN	O	O
fMet-Leu-Phe	NN	O	O
)	NN	O	O
that	NN	O	O
induce	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
chemokines	NN	O	B-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
proteins	NN	O	I-protein
translocated	NN	O	O
to	NN	O	O
nuclear	NN	O	O
fractions	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
transient	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
in	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

The	NN	O	O
onset	NN	O	O
of	NN	O	O
both	NN	O	O
processes	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
closely	NN	O	O
paralleled	NN	O	O
by	NN	O	O
,	NN	O	O
and	NN	O	O
dependent	NN	O	O
on	NN	O	O
,	NN	O	O
IkappaB-alpha	NN	O	B-protein
degradation	NN	O	O
.	NN	O	O

Proinflammatory	NN	O	B-cell_type
neutrophil	NN	O	I-cell_type
stimuli	NN	O	O
also	NN	O	O
promoted	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcripts	NN	O	I-RNA
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
reexpression	NN	O	O
of	NN	O	O
the	NN	O	O
IkappaB-alpha	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

To	NN	O	O
our	NN	O	O
knowledge	NN	O	O
,	NN	O	O
this	NN	O	O
constitutes	NN	O	O
the	NN	O	O
first	NN	O	O
indication	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
may	NN	O	O
underlie	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
proinflammatory	NN	O	O
stimuli	NN	O	O
towards	NN	O	O
human	NN	O	O
neutrophil	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
,	NN	O	O
as	NN	O	O
such	NN	O	O
,	NN	O	O
adds	NN	O	O
a	NN	O	O
new	NN	O	O
facet	NN	O	O
to	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
neutrophil	NN	O	B-cell_type
biology	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Two	NN	O	O
distinct	NN	O	O
pathways	NN	O	O
of	NN	O	O
interleukin-5	NN	O	B-protein
synthesis	NN	O	O
in	NN	O	O
allergen-specific	NN	O	B-cell_line
human	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
are	NN	O	O
suppressed	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
have	NN	O	O
long	NN	O	O
been	NN	O	O
used	NN	O	O
as	NN	O	O
the	NN	O	O
most	NN	O	O
effective	NN	O	O
agents	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
allergic	NN	O	O
diseases	NN	O	O
accompanied	NN	O	O
by	NN	O	O
eosinophilia	NN	O	O
such	NN	O	O
as	NN	O	O
chronic	NN	O	O
asthma	NN	O	O
and	NN	O	O
atopic	NN	O	O
dermatitis	NN	O	O
.	NN	O	O

The	NN	O	O
development	NN	O	O
of	NN	O	O
chronic	NN	O	O
eosinophilic	NN	O	O
inflammation	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
interleukin-5	NN	O	B-protein
(	NN	O	O
IL-5	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
selective	NN	O	B-protein
eosinophil-activating	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
produced	NN	O	O
by	NN	O	O
helper	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
delineate	NN	O	O
the	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
human	NN	O	O
IL-5	NN	O	B-protein
synthesis	NN	O	O
,	NN	O	O
we	NN	O	O
established	NN	O	O
allergen-specific	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
from	NN	O	O
asthmatic	NN	O	O
patients	NN	O	O
.	NN	O	O

GC	NN	O	O
efficiently	NN	O	O
suppressed	NN	O	O
IL-5	NN	O	B-protein
synthesis	NN	O	O
of	NN	O	O
T-cell	NN	O	O
clones	NN	O	O
activated	NN	O	O
via	NN	O	O
either	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
or	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-2R	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
IL-5	NN	O	B-RNA
mRNA	NN	O	I-RNA
upon	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
IL-2R	NN	O	B-protein
stimulation	NN	O	O
was	NN	O	O
totally	NN	O	O
inhibited	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

Human	NN	O	O
IL-5	NN	O	B-protein
promoter/enhancer-luciferase	NN	O	O
gene	NN	O	O
construct	NN	O	O
transfected	NN	O	O
to	NN	O	O
T-cell	NN	O	B-cell_line
clones	NN	O	I-cell_line
was	NN	O	O
transcribed	NN	O	O
on	NN	O	O
either	NN	O	O
TCR	NN	O	B-protein
or	NN	O	O
IL-2R	NN	O	B-protein
stimulation	NN	O	O
and	NN	O	O
was	NN	O	O
clearly	NN	O	O
downregulated	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
approximately	NN	O	O
500-bp	NN	O	B-DNA
human	NN	O	I-DNA
IL-5	NN	O	I-DNA
gene	NN	O	I-DNA
segment	NN	O	I-DNA
located	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
coding	NN	O	O
region	NN	O	O
contains	NN	O	O
activation-inducible	NN	O	B-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
regulation	NN	O	O
by	NN	O	O
GC	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
analysis	NN	O	O
suggested	NN	O	O
that	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
are	NN	O	O
among	NN	O	O
the	NN	O	O
possible	NN	O	O
targets	NN	O	O
of	NN	O	O
GC	NN	O	O
actions	NN	O	O
on	NN	O	O
TCR-stimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NF-AT	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
were	NN	O	O
not	NN	O	O
significantly	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
showing	NN	O	O
that	NN	O	O
GC	NN	O	O
suppressed	NN	O	O
IL-5	NN	O	B-protein
production	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
activated	NN	O	O
by	NN	O	O
two	NN	O	O
distinct	NN	O	O
stimuli	NN	O	O
,	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
IL-2R	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
underscore	NN	O	O
the	NN	O	O
efficacy	NN	O	O
of	NN	O	O
GC	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
allergic	NN	O	O
diseases	NN	O	O
via	NN	O	O
suppression	NN	O	O
of	NN	O	O
T-cell	NN	O	O
IL-5	NN	O	B-protein
synthesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Itk	NN	O	B-protein
,	NN	O	O
a	NN	O	O
T	NN	O	B-protein
cell-specific	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
CD2-mediated	NN	O	B-DNA
interleukin-2	NN	O	I-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
functional	NN	O	O
role	NN	O	O
of	NN	O	O
Itk	NN	O	B-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
Tec	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
either	NN	O	O
CD2	NN	O	B-protein
or	NN	O	O
T	NN	O	O
cell	NN	O	O
receptor	NN	O	O
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
/CD3	NN	O	O
on	NN	O	O
Tcells	NN	O	O
by	NN	O	O
monoclonal	NN	O	O
antibody-mediated	NN	O	O
cross-linking	NN	O	O
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Itk	NN	O	B-protein
,	NN	O	O
which	NN	O	O
was	NN	O	O
maximal	NN	O	O
as	NN	O	O
early	NN	O	O
as	NN	O	O
1	NN	O	O
min	NN	O	O
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
anti-	NN	O	O
Itk	NN	O	B-protein
immunoprecipitate	NN	O	O
was	NN	O	O
significantly	NN	O	O
activated	NN	O	O
upon	NN	O	O
these	NN	O	O
stimulations	NN	O	O
.	NN	O	O

Interleukin-2	NN	O	O
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
promoter	NN	O	O
activity	NN	O	O
stimulated	NN	O	O
by	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
,	NN	O	O
TCR	NN	O	B-protein
/CD3	NN	O	O
,	NN	O	O
and	NN	O	O
CD28	NN	O	B-protein
with	NN	O	O
antibodies	NN	O	B-protein
was	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
by	NN	O	O
transient	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
Itk	NN	O	B-protein
mutant	NN	O	I-protein
lacking	NN	O	O
the	NN	O	O
kinase	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
reduction	NN	O	O
paralleled	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
wild-type	NN	O	I-protein
Itk	NN	O	I-protein
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
or	NN	O	O
TCR/CD3	NN	O	B-protein
induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
of	NN	O	O
which	NN	O	O
is	NN	O	O
included	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
was	NN	O	O
also	NN	O	O
impaired	NN	O	O
by	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Itk	NN	O	B-protein
mutant	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Itk	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
critical	NN	O	O
involvement	NN	O	O
of	NN	O	O
Itk	NN	O	B-protein
in	NN	O	O
the	NN	O	O
initial	NN	O	O
stage	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
by	NN	O	O
mediating	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
TCR/CD3	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
CD2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
LAZ3/BCL6	NN	O	B-protein
in	NN	O	O
follicular	NN	O	O
center	NN	O	O
(	NN	O	O
FC	NN	O	O
)	NN	O	O
B	NN	O	O
cells	NN	O	O
of	NN	O	O
reactive	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
and	NN	O	O
FC-derived	NN	O	O
non-Hodgkin	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

Chromosomal	NN	O	O
translocation	NN	O	O
resulting	NN	O	O
in	NN	O	O
abnormal	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
LAZ3/BCL6	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
tumorigenesis	NN	O	O
of	NN	O	O
non-Hodgkin	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
NHL	NN	O	O
)	NN	O	O
.	NN	O	O

Therefore	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
LAZ3/BCL6	NN	O	B-protein
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
with	NN	O	O
synthetic	NN	O	O
oligonucleotide	NN	O	O
probes	NN	O	O
in	NN	O	O
frozen	NN	O	O
tissue	NN	O	O
sections	NN	O	O
from	NN	O	O
five	NN	O	O
reactive	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
and	NN	O	O
38	NN	O	O
B	NN	O	O
cell	NN	O	O
and	NN	O	O
non-B	NN	O	O
NHL	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
LAZ3/BCL6	NN	O	B-protein
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
on	NN	O	O
multiple	NN	O	O
human	NN	O	O
tissues	NN	O	O
.	NN	O	O

The	NN	O	O
LAZ3/BCL6	NN	O	B-RNA
transcript	NN	O	I-RNA
was	NN	O	O
found	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
tissues	NN	O	O
,	NN	O	O
including	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
,	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
weakly	NN	O	O
in	NN	O	O
normal	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
tumor	NN	O	O
samples	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
LAZ3/BCL6	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
in	NN	O	O
68	NN	O	O
%	NN	O	O
of	NN	O	O
all	NN	O	O
B	NN	O	O
cell	NN	O	O
NHL	NN	O	O
and	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
non-B	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

All	NN	O	O
cases	NN	O	O
of	NN	O	O
follicular	NN	O	O
,	NN	O	O
mixed	NN	O	O
small	NN	O	O
and	NN	O	O
large	NN	O	O
cell	NN	O	O
lymphomas	NN	O	O
showed	NN	O	O
LAZ3/BCL6	NN	O	B-protein
expression	NN	O	O
confined	NN	O	O
to	NN	O	O
the	NN	O	O
neoplastic	NN	O	O
follicles	NN	O	O
.	NN	O	O

A	NN	O	O
follicular	NN	O	O
expression	NN	O	O
pattern	NN	O	O
was	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
all	NN	O	O
non-malignant	NN	O	O
reactive	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
LAZ3/BCL6	NN	O	B-protein
does	NN	O	O
not	NN	O	O
correlate	NN	O	O
to	NN	O	O
malignancy	NN	O	O
,	NN	O	O
but	NN	O	O
reflects	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
germinal	NN	O	O
centers	NN	O	O
.	NN	O	O

-DOCSTART-	O

Jak1	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
mediating	NN	O	O
interleukin-4-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
insulin	NN	O	O
receptor	NN	O	O
substrate	NN	O	O
and	NN	O	O
Stat6	NN	O	B-protein
signaling	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Jak1	NN	O	B-protein
,	NN	O	I-protein
Jak2	NN	O	I-protein
,	NN	O	I-protein
Jak3	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
Fes	NN	O	I-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
have	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
undergo	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-4	NN	O	I-protein
stimulation	NN	O	O
in	NN	O	O
different	NN	O	O
cell	NN	O	O
systems	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
clear	NN	O	O
which	NN	O	O
,	NN	O	O
if	NN	O	O
any	NN	O	O
,	NN	O	O
of	NN	O	O
these	NN	O	O
kinases	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
initiating	NN	O	O
IL-4-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
intracellular	NN	O	B-protein
substrates	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
utilized	NN	O	O
a	NN	O	O
mutant	NN	O	B-cell_line
Jak1-deficient	NN	O	I-cell_line
HeLa	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
E1C3	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
its	NN	O	O
parental	NN	O	B-cell_line
Jak1-expressing	NN	O	I-cell_line
counterpart	NN	O	I-cell_line
,	NN	O	O
1D4	NN	O	B-cell_line
,	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Jak1	NN	O	B-protein
in	NN	O	O
mediating	NN	O	O
IL-4-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
events	NN	O	O
.	NN	O	O

IL-4	NN	O	O
treatment	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
insulin	NN	O	B-protein
receptor	NN	O	I-protein
substrate	NN	O	I-protein
(	NN	O	I-protein
IRS	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
and	NN	O	O
IRS-2	NN	O	B-protein
in	NN	O	O
1D4	NN	O	B-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
E1C3	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

IL-4	NN	O	B-protein
-mediated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
was	NN	O	O
pronounced	NN	O	O
in	NN	O	O
1D4	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
no	NN	O	O
IL-4	NN	O	B-protein
-induced	NN	O	O
Stat6	NN	O	B-protein
phosphorylation	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
E1C3	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

IL-4	NN	O	B-protein
also	NN	O	O
induced	NN	O	O
Stat6	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
from	NN	O	O
lysates	NN	O	O
of	NN	O	O
1D4	NN	O	B-cell_line
but	NN	O	O
not	NN	O	O
E1C3	NN	O	B-cell_line
cells	NN	O	I-cell_line
utilizing	NN	O	O
a	NN	O	O
radiolabeled	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
germline	NN	O	I-DNA
epsilon	NN	O	I-DNA
promotor	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
Iepsilon	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
an	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

Reconstitution	NN	O	O
of	NN	O	O
Jak1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
E1C3	NN	O	B-cell_line
cells	NN	O	I-cell_line
restored	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
IRS	NN	O	B-DNA
and	NN	O	O
Stat6	NN	O	B-protein
tyrosine	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
Jak1	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
mediating	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
crucial	NN	O	O
molecules	NN	O	O
involved	NN	O	O
in	NN	O	O
IL-4	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

-DOCSTART-	O

Oxidant-regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
chronically	NN	O	O
inflamed	NN	O	O
intestine	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
becoming	NN	O	O
increasingly	NN	O	O
apparent	NN	O	O
that	NN	O	O
the	NN	O	O
chronic	NN	O	O
gut	NN	O	O
inflammation	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
idiopathic	NN	O	O
inflammatory	NN	O	O
bowel	NN	O	O
diseases	NN	O	O
(	NN	O	O
e.g.	NN	O	O
ulcerative	NN	O	O
colitis	NN	O	O
,	NN	O	O
Crohn	NN	O	O
's	NN	O	O
disease	NN	O	O
)	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
enhanced	NN	O	O
production	NN	O	O
of	NN	O	O
leukocyte-derived	NN	O	O
oxidants	NN	O	O
.	NN	O	O

Oxidants	NN	O	O
such	NN	O	O
as	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
activate	NN	O	O
certain	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
beta	NN	O	I-protein
.	NN	O	O

Nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
kB	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
ubiquitous	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
pleiotropic	NN	O	B-protein
regulator	NN	O	I-protein
of	NN	O	O
numerous	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

This	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
activated	NN	O	O
via	NN	O	O
the	NN	O	O
selective	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
ubiquination	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
its	NN	O	O
inhibitor	NN	O	B-protein
protein	NN	O	I-protein
I-kB	NN	O	I-protein
thereby	NN	O	O
allowing	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
where	NN	O	O
it	NN	O	O
upregulates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
(	NN	O	O
e.g	NN	O	O
.	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
VCAM-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
IL-1	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
and	NN	O	O
enzymes	NN	O	B-protein
(	NN	O	O
iNOS	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
proteolytic	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
post-translationally	NN	O	O
modified	NN	O	O
I-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
26S	NN	O	B-protein
proteasome	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Based	NN	O	O
upon	NN	O	O
work	NN	O	O
from	NN	O	O
our	NN	O	O
laboratory	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
produces	NN	O	O
significant	NN	O	O
anti	NN	O	O
inflammatory	NN	O	O
activity	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
certain	NN	O	O
pro-inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
and	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Generation	NN	O	O
of	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
against	NN	O	O
immunorecessive	NN	O	B-protein
epitopes	NN	O	I-protein
after	NN	O	O
multiple	NN	O	O
immunizations	NN	O	O
with	NN	O	O
adenovirus	NN	O	O
vectors	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
haplotype	NN	O	O
.	NN	O	O

Currently	NN	O	O
,	NN	O	O
adenovirus	NN	O	O
(	NN	O	O
Ad	NN	O	O
)	NN	O	O
is	NN	O	O
being	NN	O	O
considered	NN	O	O
as	NN	O	O
a	NN	O	O
vector	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
cystic	NN	O	O
fibrosis	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
other	NN	O	O
diseases	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
(	NN	O	O
CTL	NN	O	B-cell_type
)	NN	O	O
response	NN	O	O
to	NN	O	O
Ad	NN	O	O
could	NN	O	O
limit	NN	O	O
the	NN	O	O
effectiveness	NN	O	O
of	NN	O	O
such	NN	O	O
approaches	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
CTL	NN	O	B-cell_type
response	NN	O	O
to	NN	O	O
virus	NN	O	O
infection	NN	O	O
is	NN	O	O
often	NN	O	O
focused	NN	O	O
on	NN	O	O
one	NN	O	O
or	NN	O	O
a	NN	O	O
few	NN	O	O
immunodominant	NN	O	B-protein
epitopes	NN	O	I-protein
,	NN	O	O
one	NN	O	O
approach	NN	O	O
to	NN	O	O
circumvent	NN	O	O
this	NN	O	O
response	NN	O	O
is	NN	O	O
to	NN	O	O
create	NN	O	O
vectors	NN	O	O
that	NN	O	O
lack	NN	O	O
these	NN	O	O
immunodominant	NN	O	B-protein
epitopes	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effectiveness	NN	O	O
of	NN	O	O
this	NN	O	O
approach	NN	O	O
was	NN	O	O
tested	NN	O	O
by	NN	O	O
immunizing	NN	O	O
mice	NN	O	O
with	NN	O	O
human	NN	O	O
group	NN	O	O
C	NN	O	O
adenoviruses	NN	O	O
.	NN	O	O

Three	NN	O	O
mouse	NN	O	O
strains	NN	O	O
(	NN	O	O
C57BL/10SnJ	NN	O	O
[	NN	O	O
H-2b	NN	O	O
]	NN	O	O
,	NN	O	O
C3HeB/FeJ	NN	O	O
[	NN	O	O
H-2k	NN	O	O
]	NN	O	O
,	NN	O	O
and	NN	O	O
BALB/cByJ	NN	O	O
[	NN	O	O
H-2d	NN	O	O
]	NN	O	O
)	NN	O	O
were	NN	O	O
immunized	NN	O	O
with	NN	O	O
wild-type	NN	O	O
Ad	NN	O	O
or	NN	O	O
Ad	NN	O	O
vectors	NN	O	O
lacking	NN	O	O
the	NN	O	O
immunodominant	NN	O	B-protein
antigen	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
CTL	NN	O	B-cell_type
responses	NN	O	O
were	NN	O	O
measured	NN	O	O
.	NN	O	O

In	NN	O	O
C57BL/10	NN	O	O
(	NN	O	O
B10	NN	O	O
)	NN	O	O
mice	NN	O	O
,	NN	O	O
a	NN	O	O
single	NN	O	O
inoculation	NN	O	O
intraperitoneally	NN	O	O
(	NN	O	O
i.p.	NN	O	O
)	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
recognition	NN	O	O
of	NN	O	O
an	NN	O	O
immunodominant	NN	O	B-protein
antigen	NN	O	I-protein
in	NN	O	O
E1A	NN	O	B-DNA
.	NN	O	O

When	NN	O	O
B10	NN	O	O
mice	NN	O	O
were	NN	O	O
inoculated	NN	O	O
multiple	NN	O	O
times	NN	O	O
either	NN	O	O
i.p	NN	O	O
.	NN	O	O

or	NN	O	O
intranasally	NN	O	O
with	NN	O	O
wild-type	NN	O	O
Ad	NN	O	O
or	NN	O	O
an	NN	O	O
Ad	NN	O	O
vector	NN	O	O
lacking	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
E1	NN	O	B-DNA
region	NN	O	I-DNA
,	NN	O	O
subdominant	NN	O	O
epitopes	NN	O	O
outside	NN	O	O
this	NN	O	O
region	NN	O	O
were	NN	O	O
recognized	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
C3H	NN	O	O
mice	NN	O	O
inoculated	NN	O	O
with	NN	O	O
wild-type	NN	O	O
Ad	NN	O	O
recognized	NN	O	O
an	NN	O	O
epitope	NN	O	O
mapping	NN	O	O
within	NN	O	O
E1B	NN	O	B-DNA
.	NN	O	O
When	NN	O	O
inoculated	NN	O	O
twice	NN	O	O
with	NN	O	O
Ad	NN	O	O
vectors	NN	O	O
lacking	NN	O	O
both	NN	O	O
E1A	NN	O	B-DNA
and	NN	O	O
E1B	NN	O	B-DNA
,	NN	O	O
no	NN	O	O
immunorecessive	NN	O	B-protein
epitopes	NN	O	I-protein
were	NN	O	O
recognized	NN	O	O
.	NN	O	O

The	NN	O	O
immune	NN	O	O
response	NN	O	O
to	NN	O	O
Ad	NN	O	O
in	NN	O	O
BALB/c	NN	O	O
mice	NN	O	O
was	NN	O	O
more	NN	O	O
complex	NN	O	O
.	NN	O	O

CTLs	NN	O	B-cell_type
from	NN	O	O
BALB/c	NN	O	O
mice	NN	O	O
inoculated	NN	O	O
i.p.	NN	O	O
with	NN	O	O
wild-type	NN	O	O
Ad	NN	O	O
recognized	NN	O	O
E1B	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
(	NN	O	I-DNA
MHC	NN	O	I-DNA
)	NN	O	I-DNA
class	NN	O	I-DNA
I	NN	O	I-DNA
Dd	NN	O	I-DNA
allele	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
region	NN	O	O
outside	NN	O	O
E1	NN	O	B-DNA
associated	NN	O	O
with	NN	O	O
the	NN	O	O
Kd	NN	O	B-DNA
allele	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
BALB/c	NN	O	O
mice	NN	O	O
were	NN	O	O
inoculated	NN	O	O
with	NN	O	O
E1	NN	O	B-DNA
-deleted	NN	O	O
Ad	NN	O	O
vectors	NN	O	O
,	NN	O	O
only	NN	O	O
the	NN	O	O
immunodominant	NN	O	B-protein
Kd-restricted	NN	O	I-protein
epitope	NN	O	I-protein
was	NN	O	O
recognized	NN	O	O
,	NN	O	O
and	NN	O	O
Dd-restricted	NN	O	O
CTLs	NN	O	B-cell_type
did	NN	O	O
not	NN	O	O
develop	NN	O	O
.	NN	O	O

This	NN	O	O
report	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
emergence	NN	O	O
of	NN	O	O
CTLs	NN	O	B-cell_type
against	NN	O	O
immunorecessive	NN	O	B-protein
epitopes	NN	O	I-protein
following	NN	O	O
multiple	NN	O	O
administrations	NN	O	O
of	NN	O	O
Ad	NN	O	O
vectors	NN	O	O
lacking	NN	O	O
immunodominant	NN	O	O
antigens	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
haplotype	NN	O	O
and	NN	O	O
could	NN	O	O
present	NN	O	O
an	NN	O	O
obstacle	NN	O	O
to	NN	O	O
gene	NN	O	O
therapy	NN	O	O
in	NN	O	O
an	NN	O	O
MHC	NN	O	B-protein
-diverse	NN	O	O
human	NN	O	O
population	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
platelet/endothelial	NN	O	I-DNA
cell	NN	O	I-DNA
adhesion	NN	O	I-DNA
molecule-1	NN	O	I-DNA
promoter	NN	O	I-DNA
:	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
GATA-2	NN	O	B-DNA
binding	NN	O	I-DNA
element	NN	O	I-DNA
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

Platelet/endothelial	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
PECAM-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
130-kD	NN	O	B-protein
member	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
gene	NN	O	I-DNA
superfamily	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
on	NN	O	O
platelets	NN	O	B-cell_type
,	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
certain	NN	O	O
leukocyte	NN	O	B-cell_type
subsets	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
factors	NN	O	O
controlling	NN	O	O
vascular-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
PECAM-1	NN	O	B-protein
,	NN	O	O
we	NN	O	O
cloned	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
PECAM-1	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
analyzed	NN	O	O
its	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

5'-Rapid	NN	O	O
amplification	NN	O	O
of	NN	O	O
cDNA	NN	O	B-DNA
ends	NN	O	I-DNA
(	NN	O	O
5'-RACE	NN	O	B-DNA
)	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
occurred	NN	O	O
at	NN	O	O
several	NN	O	O
closely	NN	O	O
spaced	NN	O	O
nearby	NN	O	O
sites	NN	O	O
originating	NN	O	O
approximately	NN	O	O
204	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
translation	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
sequence	NN	O	O
immediately	NN	O	O
upstream	NN	O	O
from	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
TIS	NN	O	B-DNA
)	NN	O	O
showed	NN	O	O
no	NN	O	O
canonical	NN	O	O
TATA	NN	O	B-DNA
or	NN	O	O
CAAT	NN	O	B-DNA
elements	NN	O	I-DNA
,	NN	O	O
however	NN	O	O
an	NN	O	O
initiator	NN	O	O
element	NN	O	O
commonly	NN	O	O
found	NN	O	O
in	NN	O	O
TATA-less	NN	O	B-DNA
promoters	NN	O	I-DNA
encompassed	NN	O	O
the	NN	O	O
TIS	NN	O	B-DNA
.	NN	O	O

5'-serially	NN	O	B-DNA
truncated	NN	O	I-DNA
PECAM-1	NN	O	I-DNA
promoter	NN	O	I-DNA
segments	NN	O	I-DNA
cloned	NN	O	O
in	NN	O	O
front	NN	O	O
of	NN	O	O
a	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
drove	NN	O	O
transcription	NN	O	O
in	NN	O	O
both	NN	O	O
a	NN	O	O
lineage-	NN	O	O
and	NN	O	O
orientation-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

Putative	NN	O	O
cis-acting	NN	O	B-DNA
control	NN	O	I-DNA
elements	NN	O	I-DNA
present	NN	O	O
within	NN	O	O
a	NN	O	O
300-bp	NN	O	B-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
included	NN	O	O
two	NN	O	O
ets	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
an	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
,	NN	O	O
tandem	NN	O	O
E-box	NN	O	B-DNA
domains	NN	O	I-DNA
,	NN	O	O
two	NN	O	O
GATA-associated	NN	O	B-DNA
sites	NN	O	I-DNA
(	NN	O	O
CACCC	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
AP-2	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
GATA	NN	O	B-DNA
element	NN	O	I-DNA
at	NN	O	O
-24	NN	O	B-DNA
.	NN	O	O

Mutational	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
optimal	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
required	NN	O	O
the	NN	O	O
GATA	NN	O	B-DNA
sequence	NN	O	I-DNA
at	NN	O	O
position	NN	O	O
-24	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
gel-shift	NN	O	O
assays	NN	O	O
further	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
GATA-2	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
bound	NN	O	O
to	NN	O	O
this	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
PECAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Understanding	NN	O	O
the	NN	O	O
cis-	NN	O	B-protein
and	NN	O	I-protein
transacting	NN	O	I-protein
factors	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
PECAM-1	NN	O	B-protein
should	NN	O	O
increase	NN	O	O
our	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
vascular-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
achieved	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
region	NN	O	O
containing	NN	O	O
a	NN	O	O
glucocorticosteroid	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
nGRE	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin-1beta	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
(	NN	O	O
IL-1beta	NN	O	B-protein
)	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
important	NN	O	O
inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
in	NN	O	O
human	NN	O	O
inflammatory	NN	O	O
and	NN	O	O
immunological	NN	O	O
diseases	NN	O	O
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	B-protein
IL-1beta	NN	O	I-protein
gene	NN	O	O
expression	NN	O	O
has	NN	O	O
been	NN	O	O
studied	NN	O	O
for	NN	O	O
several	NN	O	O
years	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
few	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
discovered	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
expression	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
anti-inflammatory	NN	O	O
and	NN	O	O
immunosuppressive	NN	O	O
effects	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
located	NN	O	O
in	NN	O	O
the	NN	O	O
region	NN	O	O
between	NN	O	O
-685	NN	O	B-DNA
and	NN	O	I-DNA
-395	NN	O	I-DNA
.	NN	O	O

Within	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
a	NN	O	O
19-bp	NN	O	B-DNA
nuclear	NN	O	I-DNA
factor	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
-570	NN	O	B-DNA
to	NN	O	I-DNA
-552	NN	O	I-DNA
)	NN	O	O
was	NN	O	O
characterized	NN	O	O
by	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
footprinting	NN	O	O
and	NN	O	O
electromobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

A	NN	O	O
consensus	NN	O	B-DNA
sequence	NN	O	I-DNA
for	NN	O	O
a	NN	O	O
negative	NN	O	B-DNA
glucocorticoid	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
nGRE	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
a	NN	O	O
transcription	NN	O	B-DNA
activator	NN	O	I-DNA
protein-2	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
were	NN	O	O
noted	NN	O	O
within	NN	O	O
this	NN	O	O
footprint	NN	O	O
.	NN	O	O

Functional	NN	O	O
studies	NN	O	O
showed	NN	O	O
a	NN	O	O
2.5-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
promoter	NN	O	O
activity	NN	O	O
when	NN	O	O
this	NN	O	O
19-bp	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
deleted	NN	O	O
in	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
constructs	NN	O	I-DNA
IL-1beta	NN	O	B-protein
/CAT	NN	O	B-protein
and	NN	O	O
IL-1beta	NN	O	B-protein
/SV40	NN	O	B-DNA
promoter	NN	O	I-DNA
/CAT	NN	O	B-protein
.	NN	O	O

Dexamethasone	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
repressed	NN	O	O
chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
(	NN	O	O
CAT	NN	O	B-protein
)	NN	O	O
production	NN	O	O
by	NN	O	O
75	NN	O	O
%	NN	O	O
in	NN	O	O
the	NN	O	O
wild-type	NN	O	B-DNA
fragment	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
in	NN	O	O
a	NN	O	O
deletion	NN	O	B-DNA
mutant	NN	O	I-DNA
lacking	NN	O	O
the	NN	O	O
19-bp	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
protein	NN	O	O
of	NN	O	O
about	NN	O	O
150	NN	O	B-protein
kD	NN	O	I-protein
that	NN	O	O
bound	NN	O	O
to	NN	O	O
this	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
sequence	NN	O	I-DNA
was	NN	O	O
identified	NN	O	O
by	NN	O	O
UV	NN	O	O
cross-linking	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
description	NN	O	O
of	NN	O	O
a	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
responsive	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
a	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Interaction	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
RFX1	NN	O	B-protein
and	NN	O	O
MIBP1	NN	O	B-protein
with	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
motif	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
hepatitis	NN	O	B-DNA
B	NN	O	I-DNA
virus	NN	O	I-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
NRE	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
hepatitis	NN	O	B-DNA
B	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
HBV	NN	O	I-DNA
)	NN	O	I-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
three	NN	O	O
subregions	NN	O	O
which	NN	O	O
act	NN	O	O
synergistically	NN	O	O
to	NN	O	O
suppress	NN	O	O
core	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
subregions	NN	O	O
,	NN	O	O
NRE	NN	O	B-DNA
gamma	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
active	NN	O	O
in	NN	O	O
both	NN	O	O
HeLa	NN	O	B-cell_line
cervical	NN	O	I-cell_line
carcinoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
Huh7	NN	O	B-cell_line
hepatoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
bound	NN	O	O
by	NN	O	O
a	NN	O	O
protein	NN	O	O
factor	NN	O	O
present	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
RFX1	NN	O	B-protein
can	NN	O	O
bind	NN	O	O
to	NN	O	O
NRE	NN	O	B-DNA
gamma	NN	O	I-DNA
and	NN	O	O
transactivate	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
promoter	NN	O	I-DNA
through	NN	O	O
this	NN	O	O
site	NN	O	O
.	NN	O	O

Mutations	NN	O	O
which	NN	O	O
abrogated	NN	O	O
the	NN	O	O
gene-suppressive	NN	O	O
activity	NN	O	O
of	NN	O	O
NRE	NN	O	B-DNA
gamma	NN	O	I-DNA
prevented	NN	O	O
RFX1	NN	O	B-protein
from	NN	O	O
binding	NN	O	O
to	NN	O	O
NRE	NN	O	B-DNA
gamma	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
RFX1	NN	O	B-protein
can	NN	O	O
bind	NN	O	O
simultaneously	NN	O	O
,	NN	O	O
most	NN	O	O
likely	NN	O	O
as	NN	O	O
a	NN	O	O
heterodimer	NN	O	B-protein
,	NN	O	O
with	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
MIBP1	NN	O	B-protein
to	NN	O	O
NRE	NN	O	B-DNA
gamma	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
cloned	NN	O	O
MIBP1	NN	O	B-DNA
gene	NN	O	I-DNA
for	NN	O	O
further	NN	O	O
studies	NN	O	O
,	NN	O	O
we	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
RFX1	NN	O	B-protein
acts	NN	O	O
with	NN	O	O
MIBP1	NN	O	B-protein
to	NN	O	O
negatively	NN	O	O
regulate	NN	O	O
the	NN	O	O
core	NN	O	O
promoter	NN	O	O
activity	NN	O	O
through	NN	O	O
the	NN	O	O
NRE	NN	O	B-DNA
gamma	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
RFX1	NN	O	B-protein
to	NN	O	O
transactivate	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
promoter	NN	O	I-DNA
raises	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
RFX1	NN	O	B-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
dual	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
HBV	NN	O	O
gene	NN	O	O
expression	NN	O	O

-DOCSTART-	O

Use	NN	O	O
of	NN	O	O
arsenic	NN	O	O
trioxide	NN	O	O
(	NN	O	O
As2O3	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
:	NN	O	O
II	NN	O	O
.	NN	O	O

Clinical	NN	O	O
efficacy	NN	O	O
and	NN	O	O
pharmacokinetics	NN	O	O
in	NN	O	O
relapsed	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
therapeutic	NN	O	O
effect	NN	O	O
of	NN	O	O
arsenic	NN	O	O
trioxide	NN	O	O
(	NN	O	O
As2O3	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
acute	NN	O	O
promyelocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
APL	NN	O	O
)	NN	O	O
was	NN	O	O
evaluated	NN	O	O
among	NN	O	O
15	NN	O	O
APL	NN	O	O
patients	NN	O	O
at	NN	O	O
relapse	NN	O	O
after	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
induced	NN	O	O
and	NN	O	O
chemotherapy	NN	O	O
maintained	NN	O	O
complete	NN	O	O
remission	NN	O	O
(	NN	O	O
CR	NN	O	O
)	NN	O	O
.	NN	O	O

As2O3	NN	O	O
was	NN	O	O
administered	NN	O	O
intravenously	NN	O	O
at	NN	O	O
the	NN	O	O
dose	NN	O	O
of	NN	O	O
10	NN	O	O
mg/d	NN	O	O
.	NN	O	O

Clinical	NN	O	O
CR	NN	O	O
was	NN	O	O
achieved	NN	O	O
in	NN	O	O
nine	NN	O	O
of	NN	O	O
10	NN	O	O
(	NN	O	O
90	NN	O	O
%	NN	O	O
)	NN	O	O
patients	NN	O	O
treated	NN	O	O
with	NN	O	O
As2O3	NN	O	O
alone	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
remaining	NN	O	O
five	NN	O	O
patients	NN	O	O
treated	NN	O	O
by	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
As2O3	NN	O	O
and	NN	O	O
low-dose	NN	O	O
chemotherapeutic	NN	O	O
drugs	NN	O	O
or	NN	O	O
ATRA	NN	O	O
.	NN	O	O

During	NN	O	O
the	NN	O	O
treatment	NN	O	O
with	NN	O	O
As2O3	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
bone	NN	O	O
marrow	NN	O	O
depression	NN	O	O
and	NN	O	O
only	NN	O	O
limited	NN	O	O
side	NN	O	O
effects	NN	O	O
were	NN	O	O
encountered	NN	O	O
.	NN	O	O

Pharmacokinetic	NN	O	O
studies	NN	O	O
,	NN	O	O
which	NN	O	O
were	NN	O	O
performed	NN	O	O
in	NN	O	O
eight	NN	O	O
patients	NN	O	O
,	NN	O	O
showed	NN	O	O
that	NN	O	O
after	NN	O	O
a	NN	O	O
peak	NN	O	O
level	NN	O	O
of	NN	O	O
5.54	NN	O	O
micromol/L	NN	O	O
to	NN	O	O
7.30	NN	O	O
micromol/L	NN	O	O
,	NN	O	O
plasma	NN	O	O
arsenic	NN	O	O
was	NN	O	O
rapidly	NN	O	O
eliminated	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
continuous	NN	O	O
administration	NN	O	O
of	NN	O	O
As2O3	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
its	NN	O	O
pharmacokinetic	NN	O	O
behaviors	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
increased	NN	O	O
amounts	NN	O	O
of	NN	O	O
arsenic	NN	O	O
appeared	NN	O	O
in	NN	O	O
the	NN	O	O
urine	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
daily	NN	O	O
excretion	NN	O	O
accounting	NN	O	O
for	NN	O	O
approximately	NN	O	O
1	NN	O	O
%	NN	O	O
to	NN	O	O
8	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
total	NN	O	O
daily	NN	O	O
dose	NN	O	O
administered	NN	O	O
.	NN	O	O

Arsenic	NN	O	O
contents	NN	O	O
in	NN	O	O
hair	NN	O	O
and	NN	O	O
nail	NN	O	O
were	NN	O	O
increased	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
peak	NN	O	O
content	NN	O	O
of	NN	O	O
arsenic	NN	O	O
could	NN	O	O
reach	NN	O	O
2.5	NN	O	O
to	NN	O	O
2.7	NN	O	O
microg/g	NN	O	O
tissue	NN	O	O
at	NN	O	O
CR	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
a	NN	O	O
decline	NN	O	O
of	NN	O	O
the	NN	O	O
arsenic	NN	O	O
content	NN	O	O
in	NN	O	O
hair	NN	O	O
and	NN	O	O
nail	NN	O	O
was	NN	O	O
observed	NN	O	O
after	NN	O	O
withdrawal	NN	O	O
of	NN	O	O
the	NN	O	O
drug	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
As2O3	NN	O	O
treatment	NN	O	O
is	NN	O	O
an	NN	O	O
effective	NN	O	O
and	NN	O	O
relatively	NN	O	O
safe	NN	O	O
drug	NN	O	O
in	NN	O	O
APL	NN	O	O
patients	NN	O	O
refractory	NN	O	O
to	NN	O	O
ATRA	NN	O	O
and	NN	O	O
conventional	NN	O	O
chemotherapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nucleolin	NN	O	B-protein
is	NN	O	O
one	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
switch	NN	O	B-protein
region	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
LR1	NN	O	B-protein
.	NN	O	O

LR1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
,	NN	O	O
sequence-specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
that	NN	O	O
regulates	NN	O	O
transcription	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

LR1	NN	O	B-protein
also	NN	O	O
binds	NN	O	O
Ig	NN	O	B-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
switch	NN	O	O
region	NN	O	O
sequences	NN	O	O
and	NN	O	O
may	NN	O	O
function	NN	O	O
in	NN	O	O
class	NN	O	O
switch	NN	O	O
recombination	NN	O	O
.	NN	O	O

LR1	NN	O	B-protein
contains	NN	O	O
two	NN	O	O
polypeptides	NN	O	O
,	NN	O	O
of	NN	O	O
106	NN	O	O
kDa	NN	O	O
and	NN	O	O
45	NN	O	O
kDa	NN	O	O
,	NN	O	O
and	NN	O	O
here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
the	NN	O	O
106-kDa	NN	O	B-protein
component	NN	O	I-protein
of	NN	O	O
LR1	NN	O	B-protein
is	NN	O	O
nucleolin	NN	O	B-protein
.	NN	O	O

This	NN	O	O
identification	NN	O	O
,	NN	O	O
initially	NN	O	O
made	NN	O	O
by	NN	O	O
microsequence	NN	O	O
analysis	NN	O	O
,	NN	O	O
was	NN	O	O
verified	NN	O	O
by	NN	O	O
showing	NN	O	O
that	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
LR1	NN	O	B-protein
-DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
increased	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
nucleolin	NN	O	B-DNA
cDNA	NN	O	I-DNA
expression	NN	O	I-DNA
construct	NN	O	I-DNA
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
LR1	NN	O	B-protein
-DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
recognized	NN	O	O
by	NN	O	O
antibodies	NN	O	O
raised	NN	O	O
against	NN	O	O
recombinant	NN	O	B-protein
human	NN	O	I-protein
nucleolin	NN	O	I-protein
;	NN	O	O
and	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
transfected	NN	O	O
with	NN	O	O
epitope-tagged	NN	O	B-DNA
nucleolin	NN	O	I-DNA
expression	NN	O	I-DNA
constructs	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
LR1-DNA	NN	O	B-protein
complex	NN	O	I-protein
was	NN	O	O
recognized	NN	O	O
by	NN	O	O
the	NN	O	O
anti-tag	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

Nucleolin	NN	O	B-protein
is	NN	O	O
an	NN	O	O
abundant	NN	O	B-protein
nucleolar	NN	O	I-protein
protein	NN	O	I-protein
which	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
rDNA	NN	O	B-DNA
transcription	NN	O	O
or	NN	O	O
organization	NN	O	O
,	NN	O	O
or	NN	O	O
rRNA	NN	O	B-RNA
processing	NN	O	O
.	NN	O	O

Homology	NN	O	O
between	NN	O	O
nucleolin	NN	O	B-protein
and	NN	O	O
histone	NN	O	B-protein
H1	NN	O	I-protein
suggests	NN	O	O
that	NN	O	O
nucleolin	NN	O	B-protein
may	NN	O	O
alter	NN	O	O
DNA	NN	O	O
organization	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
cell	NN	O	O
cycle	NN	O	O
controls	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
nucleolin	NN	O	B-protein
component	NN	O	O
of	NN	O	O
LR1	NN	O	B-protein
may	NN	O	O
therefore	NN	O	O
function	NN	O	O
to	NN	O	O
organize	NN	O	O
switch	NN	O	B-DNA
regions	NN	O	I-DNA
before	NN	O	O
,	NN	O	O
during	NN	O	O
,	NN	O	O
or	NN	O	O
after	NN	O	O
switch	NN	O	O
recombination	NN	O	O
.	NN	O	O

The	NN	O	O
demonstration	NN	O	O
that	NN	O	O
nucleolin	NN	O	B-protein
is	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
a	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
complex	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
switch	NN	O	O
region	NN	O	O
sequences	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
G-rich	NN	O	B-DNA
switch	NN	O	I-DNA
regions	NN	O	I-DNA
may	NN	O	O
have	NN	O	O
evolved	NN	O	O
from	NN	O	O
rDNA	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Glucocorticoid-resistance	NN	O	O
in	NN	O	O
peripheral-blood	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
does	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
number	NN	O	O
of	NN	O	O
affinity	NN	O	O
of	NN	O	O
glucocorticoid-receptors	NN	O	B-protein
in	NN	O	O
chronic	NN	O	O
renal	NN	O	O
failure	NN	O	O
patients	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
resistance	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
renal	NN	O	O
failure	NN	O	O
(	NN	O	O
CRF	NN	O	O
)	NN	O	O
seriously	NN	O	O
impairs	NN	O	O
successive	NN	O	O
GC	NN	O	O
therapy	NN	O	O
after	NN	O	O
renal	NN	O	O
transplantation	NN	O	O
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
GC-receptor	NN	O	O
(	NN	O	O
GC-R	NN	O	O
)	NN	O	O
parameters	NN	O	O
in	NN	O	O
peripheral-blood	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
PBMC	NN	O	B-cell_type
resistance	NN	O	O
to	NN	O	O
GC	NN	O	O
in	NN	O	O
21	NN	O	O
CRF	NN	O	O
patients	NN	O	O
and	NN	O	O
18	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
.	NN	O	O

Each	NN	O	O
subject	NN	O	O
group	NN	O	O
was	NN	O	O
divided	NN	O	O
into	NN	O	O
two	NN	O	O
subgroups	NN	O	O
according	NN	O	O
to	NN	O	O
PBMC	NN	O	B-cell_type
sensitivity	NN	O	O
to	NN	O	O
prednisolone	NN	O	O
in	NN	O	O
a	NN	O	O
mitogen	NN	O	O
assay	NN	O	O
procedure	NN	O	O
;	NN	O	O
i.e.	NN	O	O
,	NN	O	O
sensitive	NN	O	O
(	NN	O	O
IC50	NN	O	O
<	NN	O	O
381	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
and	NN	O	O
resistant	NN	O	O
(	NN	O	O
IC50	NN	O	O
>	NN	O	O
381	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
groups	NN	O	O
.	NN	O	O

In	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
,	NN	O	O
the	NN	O	O
mean	NN	O	O
GC-R	NN	O	B-protein
Bmax	NN	O	O
and	NN	O	O
Kd	NN	O	O
in	NN	O	O
quiescent	NN	O	O
PBMC	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
GC-sensitive	NN	O	O
group	NN	O	O
were	NN	O	O
2.89	NN	O	O
+/-	NN	O	O
1.23	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
and	NN	O	O
4.00	NN	O	O
+/-	NN	O	O
2.24	NN	O	O
nM	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
Bmax	NN	O	O
in	NN	O	O
these	NN	O	O
subjects	NN	O	O
significantly	NN	O	O
increased	NN	O	O
to	NN	O	O
6.61	NN	O	O
+/-	NN	O	O
2.02	NN	O	O
(	NN	O	O
257.7	NN	O	O
+/-	NN	O	O
107.8	NN	O	O
%	NN	O	O
)	NN	O	O
after	NN	O	O
24	NN	O	O
h	NN	O	O
stimulation	NN	O	O
with	NN	O	O
concanavalin	NN	O	O
A	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
Kd	NN	O	O
change	NN	O	O
was	NN	O	O
not	NN	O	O
significant	NN	O	O
.	NN	O	O

The	NN	O	O
GC-R	NN	O	B-protein
Bmax	NN	O	O
and	NN	O	O
Kd	NN	O	O
in	NN	O	O
quiescent	NN	O	O
PBMC	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
GC-resistant	NN	O	O
group	NN	O	O
were	NN	O	O
5.33	NN	O	O
+/-	NN	O	O
1.37	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
and	NN	O	O
3.20	NN	O	O
+/-	NN	O	O
1.39	NN	O	O
nM	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Both	NN	O	O
of	NN	O	O
these	NN	O	O
parameters	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
did	NN	O	O
not	NN	O	O
change	NN	O	O
significantly	NN	O	O
after	NN	O	O
mitogen	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
significant	NN	O	O
negative	NN	O	O
correlation	NN	O	O
between	NN	O	O
IC50S	NN	O	O
of	NN	O	O
prednisolone	NN	O	O
and	NN	O	O
increase-ratios	NN	O	O
(	NN	O	O
post/pre	NN	O	O
ratio	NN	O	O
)	NN	O	O
of	NN	O	O
Bmax	NN	O	O
after	NN	O	O
mitogen	NN	O	B-protein
stimulation	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
CRF	NN	O	O
patients	NN	O	O
,	NN	O	O
Bmax	NN	O	O
and	NN	O	O
Kd	NN	O	O
in	NN	O	O
quiescent	NN	O	O
PBMC	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
GC-sensitive	NN	O	O
group	NN	O	O
were	NN	O	O
6.04	NN	O	O
+/-	NN	O	O
2.35	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
and	NN	O	O
3.49	NN	O	O
+/-	NN	O	O
1.72	NN	O	O
nM	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
while	NN	O	O
those	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
GC-resistant	NN	O	O
group	NN	O	O
were	NN	O	O
5.13	NN	O	O
+/-	NN	O	O
2.31	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
and	NN	O	O
4.04	NN	O	O
+/-	NN	O	O
1.62	NN	O	O
nM	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
Bmax	NN	O	O
and	NN	O	O
Kd	NN	O	O
were	NN	O	O
not	NN	O	O
significantly	NN	O	O
changed	NN	O	O
after	NN	O	O
mitogen	NN	O	B-protein
stimulation	NN	O	O
in	NN	O	O
both	NN	O	O
subgroups	NN	O	O
of	NN	O	O
CRF	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
correlation	NN	O	O
between	NN	O	O
IC50	NN	O	O
and	NN	O	O
GC-R	NN	O	B-protein
parameters	NN	O	O
in	NN	O	O
CRF	NN	O	O
.	NN	O	O

We	NN	O	O
concluded	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
,	NN	O	O
decreased	NN	O	O
PBMC	NN	O	B-cell_type
capacity	NN	O	O
to	NN	O	O
amplify	NN	O	O
GC-R	NN	O	B-protein
numbers	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogen	NN	O	B-protein
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
GC	NN	O	O
resistance	NN	O	O
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
CRF	NN	O	O
patients	NN	O	O
the	NN	O	O
resistant	NN	O	O
mechanism	NN	O	O
is	NN	O	O
not	NN	O	O
correlated	NN	O	O
with	NN	O	O
GC-R	NN	O	B-protein
parameters	NN	O	O
.	NN	O	O

An	NN	O	O
unknown	NN	O	O
event	NN	O	O
might	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
GC-resistance	NN	O	O
of	NN	O	O
CRF	NN	O	O
.	NN	O	O

-DOCSTART-	O

Alteration	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
serine	NN	O	O
in	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
ZEBRA	NN	O	I-protein
protein	NN	O	I-protein
separates	NN	O	O
its	NN	O	O
functions	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
and	NN	O	O
disruption	NN	O	O
of	NN	O	O
latency	NN	O	O
.	NN	O	O

The	NN	O	O
ZEBRA	NN	O	B-protein
protein	NN	O	I-protein
from	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
activates	NN	O	O
a	NN	O	O
switch	NN	O	O
from	NN	O	O
the	NN	O	O
latent	NN	O	O
to	NN	O	O
the	NN	O	O
lytic	NN	O	O
expression	NN	O	O
program	NN	O	O
of	NN	O	O
the	NN	O	O
virus	NN	O	O
.	NN	O	O

ZEBRA	NN	O	B-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
bZIP	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
is	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
expression	NN	O	O
from	NN	O	O
viral	NN	O	B-DNA
lytic	NN	O	I-DNA
cycle	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
had	NN	O	O
previously	NN	O	O
been	NN	O	O
thought	NN	O	O
that	NN	O	O
ZEBRA	NN	O	B-protein
's	NN	O	O
capacity	NN	O	O
to	NN	O	O
disrupt	NN	O	O
EBV	NN	O	O
latency	NN	O	O
resided	NN	O	O
primarily	NN	O	O
in	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
that	NN	O	O
encode	NN	O	O
products	NN	O	O
required	NN	O	O
for	NN	O	O
lytic	NN	O	O
replication	NN	O	O
.	NN	O	O

We	NN	O	O
generated	NN	O	O
a	NN	O	O
point	NN	O	O
mutant	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-protein
,	NN	O	O
Z	NN	O	B-protein
(	NN	O	I-protein
S186A	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
that	NN	O	O
was	NN	O	O
not	NN	O	O
impaired	NN	O	O
in	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
this	NN	O	O
mutation	NN	O	O
abolished	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
initiate	NN	O	O
the	NN	O	O
viral	NN	O	O
lytic	NN	O	O
cascade	NN	O	O
.	NN	O	O

The	NN	O	O
mutant	NN	O	O
,	NN	O	O
containing	NN	O	O
a	NN	O	O
serine-to-alanine	NN	O	O
substitution	NN	O	O
in	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
protein	NN	O	O
,	NN	O	O
bound	NN	O	O
to	NN	O	O
several	NN	O	O
known	NN	O	O
ZEBRA-binding	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
activated	NN	O	O
transcription	NN	O	O
from	NN	O	O
reporters	NN	O	O
bearing	NN	O	O
known	NN	O	O
ZEBRA-responsive	NN	O	B-DNA
promoters	NN	O	I-DNA
but	NN	O	O
did	NN	O	O
not	NN	O	O
disrupt	NN	O	O
latency	NN	O	O
in	NN	O	O
EBV-infected	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
initiation	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
by	NN	O	O
the	NN	O	O
ZEBRA	NN	O	B-protein
protein	NN	O	I-protein
requires	NN	O	O
a	NN	O	O
function	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
;	NN	O	O
a	NN	O	O
change	NN	O	O
of	NN	O	O
serine	NN	O	O
186	NN	O	O
to	NN	O	O
alanine	NN	O	O
in	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
ZEBRA	NN	O	B-protein
abolished	NN	O	O
this	NN	O	O
additional	NN	O	O
function	NN	O	O
and	NN	O	O
uncovered	NN	O	O
a	NN	O	O
new	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
ZEBRA	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
disruption	NN	O	O
of	NN	O	O
EBV	NN	O	O
latency	NN	O	O
.	NN	O	O

The	NN	O	O
additional	NN	O	O
function	NN	O	O
that	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
initiation	NN	O	O
of	NN	O	O
the	NN	O	O
lytic	NN	O	O
viral	NN	O	O
life	NN	O	O
cycle	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
require	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
serine	NN	O	O
186	NN	O	O
of	NN	O	O
the	NN	O	O
ZEBRA	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
may	NN	O	O
influence	NN	O	O
either	NN	O	O
DNA	NN	O	O
recognition	NN	O	O
or	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
lytic	NN	O	B-DNA
viral	NN	O	I-DNA
promoters	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
chromatinized	NN	O	B-DNA
viral	NN	O	I-DNA
episome	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Immune	NN	O	O
hyperactivation	NN	O	O
of	NN	O	O
HIV-1-infected	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
mediated	NN	O	O
by	NN	O	O
Tat	NN	O	B-protein
and	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus-type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
infection	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
chronic	NN	O	O
state	NN	O	O
of	NN	O	O
immune	NN	O	O
hyperactivation	NN	O	O
in	NN	O	O
patients	NN	O	O
.	NN	O	O

Infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
vitro	NN	O	O
resulted	NN	O	O
in	NN	O	O
increased	NN	O	O
interleukin-2	NN	O	O
(	NN	O	O
IL-2	NN	O	O
)	NN	O	O
secretion	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
via	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
and	NN	O	I-protein
CD28	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-protein
transactivator	NN	O	I-protein
Tat	NN	O	B-protein
recapitulated	NN	O	O
this	NN	O	O
phenotype	NN	O	O
and	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
increased	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
costimulation	NN	O	O
with	NN	O	O
CD3	NN	O	B-protein
plus	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

IL-2	NN	O	B-protein
superinduction	NN	O	O
by	NN	O	O
Tat	NN	O	B-protein
occurred	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
,	NN	O	O
was	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
CD28-responsive	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
was	NN	O	O
exclusively	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
29	NN	O	B-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
second	NN	O	B-DNA
exon	NN	O	I-DNA
of	NN	O	I-DNA
Tat	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Response	NN	O	O
to	NN	O	O
intranasal	NN	O	O
fluticasone	NN	O	O
propionate	NN	O	O
in	NN	O	O
perennial	NN	O	O
allergic	NN	O	O
rhinitis	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
characteristics	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
The	NN	O	O
reduction	NN	O	O
of	NN	O	O
symptoms	NN	O	O
due	NN	O	O
to	NN	O	O
treatment	NN	O	O
with	NN	O	O
corticosteroids	NN	O	O
varies	NN	O	O
among	NN	O	O
patients	NN	O	O
with	NN	O	O
perennial	NN	O	O
rhinitis	NN	O	O
.	NN	O	O

Most	NN	O	O
patients	NN	O	O
will	NN	O	O
respond	NN	O	O
but	NN	O	O
a	NN	O	O
few	NN	O	O
patients	NN	O	O
respond	NN	O	O
less	NN	O	O
to	NN	O	O
these	NN	O	O
drugs	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
To	NN	O	O
investigate	NN	O	O
the	NN	O	O
association	NN	O	O
in	NN	O	O
reduction	NN	O	O
of	NN	O	O
symptoms	NN	O	O
due	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
characteristics	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
perennial	NN	O	O
allergic	NN	O	O
rhinitis	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
studies	NN	O	O
were	NN	O	O
performed	NN	O	O
with	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
production	NN	O	O
of	NN	O	O
mediators	NN	O	B-protein
were	NN	O	O
measured	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
During	NN	O	O
a	NN	O	O
double-blind	NN	O	O
placebo-controlled	NN	O	O
crossover	NN	O	O
study	NN	O	O
,	NN	O	O
200	NN	O	O
micrograms	NN	O	O
fluticasone	NN	O	O
propionate	NN	O	O
aqueous	NN	O	O
nasal	NN	O	O
spray	NN	O	O
(	NN	O	O
in	NN	O	O
the	NN	O	O
active	NN	O	O
treatment	NN	O	O
period	NN	O	O
)	NN	O	O
and	NN	O	O
placebo	NN	O	O
(	NN	O	O
in	NN	O	O
the	NN	O	O
placebo	NN	O	O
treatment	NN	O	O
period	NN	O	O
)	NN	O	O
were	NN	O	O
administered	NN	O	O
twice	NN	O	O
daily	NN	O	O
for	NN	O	O
2	NN	O	O
weeks	NN	O	O
to	NN	O	O
22	NN	O	O
patients	NN	O	O
allergic	NN	O	O
to	NN	O	O
house	NN	O	O
dust	NN	O	O
mite	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
both	NN	O	O
treatment	NN	O	O
periods	NN	O	O
symptoms	NN	O	O
were	NN	O	O
scored	NN	O	O
after	NN	O	O
allergen	NN	O	O
provocation	NN	O	O
(	NN	O	O
100	NN	O	O
,	NN	O	O
1000	NN	O	O
,	NN	O	O
10000	NN	O	O
BU/mL	NN	O	O
)	NN	O	O
and	NN	O	O
during	NN	O	O
the	NN	O	O
9.5	NN	O	O
hours	NN	O	O
after	NN	O	O
this	NN	O	O
challenge	NN	O	O
.	NN	O	O

Receptor	NN	O	O
binding	NN	O	O
studies	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
were	NN	O	O
performed	NN	O	O
with	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Leukotriene	NN	O	O
B4	NN	O	O
produced	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
soluble	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
released	NN	O	O
by	NN	O	O
lymphocytes	NN	O	B-cell_type
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
in	NN	O	O
plasma	NN	O	O
were	NN	O	O
determined	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
No	NN	O	O
significant	NN	O	O
partial	NN	O	O
correlations	NN	O	O
of	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
the	NN	O	O
peripheral	NN	O	B-protein
blood	NN	O	I-protein
mononuclear	NN	O	I-protein
cell	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
6821	NN	O	O
+/-	NN	O	O
5669	NN	O	O
binding	NN	O	O
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
affinity	NN	O	O
(	NN	O	O
Kd	NN	O	O
:	NN	O	O
16.5	NN	O	O
+/-	NN	O	O
13.51	NN	O	O
nmol/L	NN	O	O
)	NN	O	O
for	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
with	NN	O	O
the	NN	O	O
symptom	NN	O	O
score	NN	O	O
(	NN	O	O
placebo	NN	O	O
:	NN	O	O
4.3	NN	O	O
+/-	NN	O	O
2.45	NN	O	O
pts	NN	O	O
;	NN	O	O
fluticasone	NN	O	O
:	NN	O	O
2.4	NN	O	O
+/-	NN	O	O
1.55	NN	O	O
pts	NN	O	O
)	NN	O	O
after	NN	O	O
active	NN	O	O
treatment	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

Also	NN	O	O
no	NN	O	O
significant	NN	O	O
partial	NN	O	O
correlations	NN	O	O
of	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
leukotriene	NN	O	O
B4	NN	O	O
(	NN	O	O
45.6	NN	O	O
+/-	NN	O	O
105.3	NN	O	O
ng/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
)	NN	O	O
produced	NN	O	O
by	NN	O	O
monocytes	NN	O	B-cell_type
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
soluble	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
734	NN	O	O
+/-	NN	O	O
237	NN	O	O
ng/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells	NN	O	O
)	NN	O	O
released	NN	O	O
by	NN	O	O
lymphocytes	NN	O	B-cell_type
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
(	NN	O	O
571	NN	O	O
+/-	NN	O	O
236	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
in	NN	O	O
plasma	NN	O	O
with	NN	O	O
the	NN	O	O
symptom	NN	O	O
score	NN	O	O
after	NN	O	O
active	NN	O	O
treatment	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
The	NN	O	O
reduction	NN	O	O
of	NN	O	O
symptoms	NN	O	O
due	NN	O	O
to	NN	O	O
topical	NN	O	O
fluticasone	NN	O	O
propionate	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
rhinitis	NN	O	O
and	NN	O	O
allergy	NN	O	O
to	NN	O	O
house	NN	O	O
dust	NN	O	O
mite	NN	O	O
is	NN	O	O
not	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
characteristics	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
homodimer	NN	O	I-protein
in	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
decreases	NN	O	O
with	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
differentiation	NN	O	O
:	NN	O	O
a	NN	O	O
possible	NN	O	O
mechanism	NN	O	O
influencing	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
replication	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
mature	NN	O	O
macrophages	NN	O	O
.	NN	O	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
replicates	NN	O	O
more	NN	O	O
efficiently	NN	O	O
in	NN	O	O
vitro	NN	O	O
in	NN	O	O
differentiated	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
than	NN	O	O
in	NN	O	O
freshly	NN	O	B-cell_type
isolated	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
whether	NN	O	O
this	NN	O	O
may	NN	O	O
be	NN	O	O
partly	NN	O	O
explained	NN	O	O
by	NN	O	O
changes	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
with	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
changed	NN	O	O
significantly	NN	O	O
with	NN	O	O
differentiation	NN	O	O
in	NN	O	O
vitro	NN	O	O
to	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
(	NN	O	O
MDMs	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
differentiation	NN	O	O
in	NN	O	O
vivo	NN	O	O
to	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
(	NN	O	O
AMs	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

Freshly	NN	O	O
isolated	NN	O	O
monocytes	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
transcriptionally	NN	O	B-protein
inactive	NN	O	I-protein
p50	NN	O	I-protein
homodimer	NN	O	I-protein
which	NN	O	O
decreased	NN	O	O
with	NN	O	O
time	NN	O	O
in	NN	O	O
culture	NN	O	O
in	NN	O	O
favor	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
p50/p65	NN	O	B-protein
and	NN	O	I-protein
p50/RelB	NN	O	I-protein
heterodimers	NN	O	I-protein
.	NN	O	O

As	NN	O	O
in	NN	O	O
MDMs	NN	O	B-cell_type
,	NN	O	O
AMs	NN	O	B-cell_type
constitutively	NN	O	O
expressed	NN	O	O
p50/p65	NN	O	B-protein
and	NN	O	O
p50/RelB	NN	O	B-protein
although	NN	O	O
at	NN	O	O
lower	NN	O	O
levels	NN	O	O
.	NN	O	O

HIV	NN	O	O
infection	NN	O	O
of	NN	O	O
fresh	NN	O	O
monocytes	NN	O	B-cell_type
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
p50/p65	NN	O	B-protein
as	NN	O	O
seen	NN	O	O
in	NN	O	O
MDMs	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
replacement	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	I-protein
with	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
heterodimers	NN	O	O
following	NN	O	O
time	NN	O	O
in	NN	O	O
culture	NN	O	O
may	NN	O	O
partially	NN	O	O
explain	NN	O	O
the	NN	O	O
progressive	NN	O	O
increase	NN	O	O
in	NN	O	O
susceptibility	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
to	NN	O	O
HIV	NN	O	O
infection	NN	O	O
during	NN	O	O
in	NN	O	O
vitro	NN	O	O
culture	NN	O	O
.	NN	O	O

The	NN	O	O
change	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
components	NN	O	I-protein
with	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
in	NN	O	O
vivo	NN	O	O
may	NN	O	O
also	NN	O	O
explain	NN	O	O
the	NN	O	O
different	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
of	NN	O	O
these	NN	O	O
cell	NN	O	O
populations	NN	O	O
in	NN	O	O
HIV-infected	NN	O	O
individuals	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-independent	NN	O	O
suppression	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
by	NN	O	O
ascorbic	NN	O	O
acid	NN	O	O
.	NN	O	O

Ascorbic	NN	O	O
acid	NN	O	O
(	NN	O	O
ascorbate	NN	O	O
or	NN	O	O
vitamin	NN	O	O
C	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
suppress	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV	NN	O	O
in	NN	O	O
latently	NN	O	B-cell_type
infected	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
following	NN	O	O
stimulation	NN	O	O
with	NN	O	O
a	NN	O	O
tumor	NN	O	O
promoter	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
inflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

To	NN	O	O
assess	NN	O	O
whether	NN	O	O
this	NN	O	O
inhibition	NN	O	O
was	NN	O	O
mediated	NN	O	O
via	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
we	NN	O	O
carried	NN	O	O
out	NN	O	O
gel	NN	O	O
shift	NN	O	O
analysis	NN	O	O
on	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
under	NN	O	O
different	NN	O	O
conditions	NN	O	O
of	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
ascorbate	NN	O	O
,	NN	O	O
N-acetylcysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
zidovudine	NN	O	O
(	NN	O	O
AZT	NN	O	O
)	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
ACH-2	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
NAC	NN	O	O
followed	NN	O	O
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
PMA	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
or	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
(	NN	O	O
H2O2	NN	O	O
)	NN	O	O
resulted	NN	O	O
in	NN	O	O
strong	NN	O	O
suppression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
neither	NN	O	O
ascorbate	NN	O	O
nor	NN	O	O
AZT	NN	O	O
affected	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
under	NN	O	O
all	NN	O	O
three	NN	O	O
induction	NN	O	O
conditions	NN	O	O
in	NN	O	O
the	NN	O	O
ACH-2	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Ascorbate	NN	O	O
and	NN	O	O
AZT	NN	O	O
also	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
following	NN	O	O
TNF-alpha-	NN	O	O
or	NN	O	O
PMA-induced	NN	O	O
stimulation	NN	O	O
of	NN	O	O
U1	NN	O	O
promonocytic	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
HIV	NN	O	O
inhibition	NN	O	O
by	NN	O	O
ascorbate	NN	O	O
is	NN	O	O
not	NN	O	O
mediated	NN	O	O
via	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibition	NN	O	O
,	NN	O	O
unlike	NN	O	O
that	NN	O	O
seen	NN	O	O
with	NN	O	O
other	NN	O	O
antioxidants	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
T	NN	O	B-DNA
cell-specific	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
interleukin-3	NN	O	B-DNA
locus	NN	O	I-DNA
is	NN	O	O
activated	NN	O	O
cooperatively	NN	O	O
by	NN	O	O
Oct	NN	O	B-DNA
and	NN	O	I-DNA
NFAT	NN	O	I-DNA
elements	NN	O	I-DNA
within	NN	O	O
a	NN	O	O
DNase	NN	O	B-DNA
I-hypersensitive	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Interleukin-3	NN	O	B-protein
(	NN	O	O
IL-3	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
primarily	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
identified	NN	O	O
an	NN	O	O
inducible	NN	O	O
T	NN	O	B-DNA
cell-specific	NN	O	I-DNA
enhancer	NN	O	I-DNA
14	NN	O	O
kb	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
responded	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

The	NN	O	O
IL-3	NN	O	B-protein
enhancer	NN	O	O
spanned	NN	O	O
an	NN	O	O
inducible	NN	O	O
cyclosporin	NN	O	B-DNA
A-sensitive	NN	O	I-DNA
DNase	NN	O	I-DNA
I-hypersensitive	NN	O	I-DNA
site	NN	O	I-DNA
found	NN	O	O
only	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Four	NN	O	O
NFAT-like	NN	O	B-DNA
elements	NN	O	I-DNA
exist	NN	O	O
within	NN	O	O
the	NN	O	O
enhancer	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
most	NN	O	O
active	NN	O	O
NFAT-like	NN	O	B-DNA
elements	NN	O	I-DNA
were	NN	O	O
located	NN	O	O
at	NN	O	O
the	NN	O	O
center	NN	O	O
of	NN	O	O
the	NN	O	O
DNase	NN	O	B-DNA
I-hypersensitive	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
NFAT-like	NN	O	B-DNA
elements	NN	O	I-DNA
encompassed	NN	O	O
overlapping	NN	O	O
Oct-	NN	O	B-DNA
and	NN	O	I-DNA
NFATp/c-binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
functioned	NN	O	O
in	NN	O	O
a	NN	O	O
highly	NN	O	O
synergistic	NN	O	O
manner	NN	O	O
.	NN	O	O

We	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
T	NN	O	O
cell-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
partly	NN	O	O
controlled	NN	O	O
through	NN	O	O
the	NN	O	O
enhancer	NN	O	O
by	NN	O	O
cooperation	NN	O	O
between	NN	O	O
Oct	NN	O	B-protein
and	NN	O	I-protein
NFAT	NN	O	I-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Adenovirus	NN	O	B-protein
E1B	NN	O	I-protein
19K	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
efficient	NN	O	O
DNA	NN	O	O
replication	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
adenovirus	NN	O	B-DNA
E1B	NN	O	I-DNA
19K	NN	O	I-DNA
gene	NN	O	I-DNA
plays	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
in	NN	O	O
transformation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
rodent	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
cooperation	NN	O	O
with	NN	O	O
E1A	NN	O	B-protein
and	NN	O	O
in	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
during	NN	O	O
lytic	NN	O	O
infection	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
that	NN	O	O
this	NN	O	O
E1B	NN	O	B-protein
19K	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
not	NN	O	O
necessary	NN	O	O
for	NN	O	O
viral	NN	O	O
DNA	NN	O	O
replication	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
such	NN	O	O
as	NN	O	O
HeLa	NN	O	B-cell_line
and	NN	O	O
KB	NN	O	B-cell_line
.	NN	O	O

We	NN	O	O
reported	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
E1B	NN	O	B-protein
19K	NN	O	I-protein
mutant	NN	O	O
viruses	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
replicate	NN	O	O
efficiently	NN	O	O
in	NN	O	O
a	NN	O	O
monocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
U937	NN	O	B-cell_line
.	NN	O	O

Viral	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
and	NN	O	O
late	NN	O	O
gene	NN	O	O
expression	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
defective	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
infected	NN	O	O
with	NN	O	O
E1B	NN	O	B-protein
19K	NN	O	I-protein
mutants	NN	O	O
compared	NN	O	O
with	NN	O	O
wild-type	NN	O	O
virus	NN	O	O
.	NN	O	O

Early	NN	O	O
viral	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
patterns	NN	O	O
also	NN	O	O
differ	NN	O	O
between	NN	O	O
wild-type	NN	O	B-cell_line
and	NN	O	I-cell_line
dl337-infected	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
defect	NN	O	O
in	NN	O	O
viral	NN	O	O
replication	NN	O	O
could	NN	O	O
be	NN	O	O
complemented	NN	O	O
by	NN	O	O
dl312	NN	O	O
virus	NN	O	O
defective	NN	O	O
in	NN	O	O
E1A	NN	O	B-protein
expression	NN	O	O
4	NN	O	O
days	NN	O	O
after	NN	O	O
infection	NN	O	O
with	NN	O	O
E1B	NN	O	O
mutants	NN	O	O
,	NN	O	O
suggesting	NN	O	O
persistence	NN	O	O
of	NN	O	O
the	NN	O	O
E1B	NN	O	B-DNA
mutant	NN	O	I-DNA
genome	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
infected	NN	O	O
cells	NN	O	O
despite	NN	O	O
defective	NN	O	O
onset	NN	O	O
of	NN	O	O
the	NN	O	O
late	NN	O	O
phase	NN	O	O
of	NN	O	O
replication	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
imply	NN	O	O
that	NN	O	O
E1B	NN	O	B-protein
19K	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
efficient	NN	O	O
viral	NN	O	O
DNA	NN	O	O
replication	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Inefficient	NN	O	O
DNA	NN	O	O
replication	NN	O	O
is	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
another	NN	O	O
monocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
Ras	NN	O	O
and	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
pathway	NN	O	O
by	NN	O	O
terminal	NN	O	B-protein
complement	NN	O	I-protein
complexes	NN	O	I-protein
is	NN	O	O
G	NN	O	B-protein
protein	NN	O	I-protein
dependent	NN	O	O
.	NN	O	O

Assembly	NN	O	O
of	NN	O	O
terminal	NN	O	B-protein
complement	NN	O	I-protein
complexes	NN	O	I-protein
(	NN	O	O
TCC	NN	O	B-protein
)	NN	O	O
C5b-7	NN	O	B-protein
,	NN	O	O
C5b-8	NN	O	B-protein
,	NN	O	O
and	NN	O	O
C5b-9	NN	O	B-protein
on	NN	O	O
target	NN	O	O
cells	NN	O	O
during	NN	O	O
acute	NN	O	O
and	NN	O	O
chronic	NN	O	O
inflammation	NN	O	O
induces	NN	O	O
hydrolysis	NN	O	O
of	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
phospholipids	NN	O	O
and	NN	O	O
heterotrimeric	NN	O	O
G	NN	O	B-protein
protein	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

TCC	NN	O	B-protein
also	NN	O	O
stimulate	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cellular	NN	O	O
activities	NN	O	O
,	NN	O	O
which	NN	O	O
include	NN	O	O
cytokine	NN	O	B-protein
synthesis	NN	O	O
,	NN	O	O
proto-oncogene	NN	O	B-DNA
activation	NN	O	O
,	NN	O	O
and	NN	O	O
mitotic	NN	O	O
signaling	NN	O	O
.	NN	O	O

Now	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
sublytic	NN	O	O
TCC	NN	O	B-protein
induced	NN	O	O
Ras	NN	O	B-protein
,	NN	O	O
Raf-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
extracellular	NN	O	B-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
ERK	NN	O	I-protein
)	NN	O	I-protein
1	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
JY25	NN	O	O
B	NN	O	O
cell	NN	O	O
line	NN	O	O
.	NN	O	O

When	NN	O	O
cells	NN	O	O
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
C5b-9	NN	O	B-protein
,	NN	O	O
GTP-bound	NN	O	B-protein
Ras	NN	O	I-protein
in	NN	O	O
anti-C5b-9	NN	O	B-protein
immunoprecipitates	NN	O	I-protein
was	NN	O	O
increased	NN	O	O
3.2-fold	NN	O	O
at	NN	O	O
2	NN	O	O
min	NN	O	O
,	NN	O	O
while	NN	O	O
GTP-bound	NN	O	B-protein
Ras	NN	O	I-protein
in	NN	O	O
anti-Ras	NN	O	B-protein
immunoprecipitates	NN	O	I-protein
was	NN	O	O
increased	NN	O	O
2-fold	NN	O	O
at	NN	O	O
10	NN	O	O
min	NN	O	O
.	NN	O	O

Both	NN	O	O
C5b-9	NN	O	B-protein
and	NN	O	O
C5b-7	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
C5b6	NN	O	B-protein
,	NN	O	O
increased	NN	O	O
Raf-1	NN	O	B-protein
kinase	NN	O	O
activity	NN	O	O
maximum	NN	O	O
3.3-fold	NN	O	O
at	NN	O	O
2	NN	O	O
min	NN	O	O
and	NN	O	O
2.8-fold	NN	O	O
at	NN	O	O
5	NN	O	O
min	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

ERK1	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
2-fold	NN	O	O
increased	NN	O	O
by	NN	O	O
C5b-9	NN	O	B-protein
at	NN	O	O
2	NN	O	O
min	NN	O	O
and	NN	O	O
by	NN	O	O
C5b-7	NN	O	B-protein
at	NN	O	O
10	NN	O	O
min	NN	O	O
,	NN	O	O
over	NN	O	O
the	NN	O	O
C5b6	NN	O	B-protein
level	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
pathway	NN	O	O
on	NN	O	O
TCC	NN	O	B-protein
-inducible	NN	O	O
mitotic	NN	O	O
signaling	NN	O	O
was	NN	O	O
evaluated	NN	O	O
by	NN	O	O
assessing	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
and	NN	O	O
activator	NN	O	B-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
MAPK/ERK-specific	NN	O	O
inhibitor	NN	O	O
PD	NN	O	O
098	NN	O	O
,	NN	O	O
059	NN	O	O
abolished	NN	O	O
the	NN	O	O
C5b-9	NN	O	B-protein
-induced	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Involvement	NN	O	O
of	NN	O	O
G	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
pathway	NN	O	O
by	NN	O	O
TCC	NN	O	B-protein
was	NN	O	O
indicated	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Raf-1	NN	O	B-protein
and	NN	O	O
ERK1	NN	O	B-protein
kinase	NN	O	O
activity	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
pertussis	NN	O	B-protein
toxin	NN	O	I-protein
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
beta-adrenergic	NN	O	O
receptor	NN	O	O
kinase	NN	O	O
1	NN	O	O
carboxyl-terminal	NN	O	O
peptide	NN	O	O
in	NN	O	O
JY25	NN	O	B-cell_line
cells	NN	O	I-cell_line
also	NN	O	O
inhibited	NN	O	O
Raf-1	NN	O	B-protein
and	NN	O	O
ERK1	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
direct	NN	O	O
involvement	NN	O	O
of	NN	O	O
G	NN	O	O
betagamma	NN	O	O
subunits	NN	O	O
in	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
generated	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
pathway	NN	O	O
by	NN	O	O
TCC	NN	O	B-protein
assembly	NN	O	O
in	NN	O	O
the	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
.	NN	O	O

-DOCSTART-	O

Spontaneous	NN	O	O
occurrence	NN	O	O
of	NN	O	O
early	NN	O	O
region	NN	O	O
1A	NN	O	O
reiteration	NN	O	O
mutants	NN	O	O
of	NN	O	O
type	NN	O	O
5	NN	O	O
adenovirus	NN	O	O
in	NN	O	O
persistently	NN	O	B-cell_type
infected	NN	O	I-cell_type
human	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
.	NN	O	O

Mutants	NN	O	O
of	NN	O	O
type	NN	O	O
5	NN	O	O
adenovirus	NN	O	O
(	NN	O	O
Ad5	NN	O	O
)	NN	O	O
with	NN	O	O
reiterated	NN	O	B-DNA
DNA	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
E1a	NN	O	B-DNA
region	NN	O	I-DNA
appeared	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T-lymphocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
Molt-4	NN	O	B-cell_line
,	NN	O	O
persistently	NN	O	O
infected	NN	O	O
with	NN	O	O
H5sub304	NN	O	O
,	NN	O	O
a	NN	O	O
deletion/substitution	NN	O	O
mutant	NN	O	O
that	NN	O	O
has	NN	O	O
a	NN	O	O
wild-type	NN	O	O
phenotype	NN	O	O
in	NN	O	O
viral	NN	O	O
replication	NN	O	O
.	NN	O	O

Endonuclease	NN	O	B-protein
analyses	NN	O	O
and	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
revealed	NN	O	O
DNA	NN	O	O
reiteration	NN	O	O
in	NN	O	O
each	NN	O	O
mutant	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
four	NN	O	O
representative	NN	O	O
mutants	NN	O	O
investigated	NN	O	O
,	NN	O	O
the	NN	O	O
DNA	NN	O	B-DNA
reiterations	NN	O	I-DNA
all	NN	O	O
started	NN	O	O
within	NN	O	O
a	NN	O	O
six-base-pair	NN	O	B-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
,	NN	O	O
G	NN	O	O
(	NN	O	O
or	NN	O	O
C	NN	O	O
)	NN	O	O
CTGTG	NN	O	O
,	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
exon	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
E1a	NN	O	B-DNA
region	NN	O	I-DNA
(	NN	O	O
at	NN	O	O
nt	NN	O	O
1333	NN	O	O
,	NN	O	O
1367	NN	O	O
,	NN	O	O
or	NN	O	O
1419	NN	O	O
)	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
not	NN	O	O
any	NN	O	O
DNA	NN	O	O
homology	NN	O	O
between	NN	O	O
the	NN	O	O
breakpoints	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
exon	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
inserting	NN	O	B-DNA
sequences	NN	O	I-DNA
(	NN	O	O
starting	NN	O	O
at	NN	O	O
nt	NN	O	O
532	NN	O	O
,	NN	O	O
710	NN	O	O
,	NN	O	O
or	NN	O	O
792	NN	O	O
)	NN	O	O
.	NN	O	O

Northern	NN	O	O
analyses	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
reiterated	NN	O	B-DNA
splicing	NN	O	I-DNA
sites	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
representative	NN	O	O
mutants	NN	O	O
were	NN	O	O
all	NN	O	O
used	NN	O	O
in	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
closest	NN	O	O
donor	NN	O	O
and	NN	O	O
recipient	NN	O	O
joints	NN	O	O
were	NN	O	O
used	NN	O	O
most	NN	O	O
frequently	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
imply	NN	O	O
that	NN	O	O
during	NN	O	O
persistent	NN	O	O
infection	NN	O	O
Ad5	NN	O	O
underwent	NN	O	O
spontaneous	NN	O	O
mutations	NN	O	O
by	NN	O	O
sequence-specific	NN	O	O
breakage	NN	O	O
and	NN	O	O
nonhomologous	NN	O	O
end-end	NN	O	O
joining	NN	O	O
recombination	NN	O	O
events	NN	O	O
.	NN	O	O

These	NN	O	O
E1a	NN	O	O
reiteration	NN	O	O
mutants	NN	O	O
could	NN	O	O
be	NN	O	O
propagated	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
,	NN	O	O
A549	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
KB	NN	O	B-cell_line
cells	NN	O	I-cell_line
;	NN	O	O
they	NN	O	O
were	NN	O	O
genetically	NN	O	O
stable	NN	O	O
;	NN	O	O
and	NN	O	O
they	NN	O	O
killed	NN	O	O
CREF	NN	O	B-cell_line
cells	NN	O	I-cell_line
at	NN	O	O
a	NN	O	O
strikingly	NN	O	O
high	NN	O	O
frequency	NN	O	O
.	NN	O	O

Preliminary	NN	O	O
observations	NN	O	O
tend	NN	O	O
to	NN	O	O
correlate	NN	O	O
this	NN	O	O
CREF	NN	O	O
cell	NN	O	O
killing	NN	O	O
with	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
early	NN	O	B-protein
viral	NN	O	I-protein
proteins	NN	O	I-protein
and/or	NN	O	O
viral	NN	O	B-DNA
DNA	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
degree	NN	O	O
of	NN	O	O
cell	NN	O	O
damage	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
Ad5wt	NN	O	O
or	NN	O	O
H5sub304	NN	O	O
infection	NN	O	O
of	NN	O	O
CREF	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
observed	NN	O	O
E1a	NN	O	B-DNA
reiterations	NN	O	I-DNA
provide	NN	O	O
a	NN	O	O
model	NN	O	O
to	NN	O	O
gain	NN	O	O
insight	NN	O	O
into	NN	O	O
understanding	NN	O	O
the	NN	O	O
evolutionary	NN	O	O
events	NN	O	O
of	NN	O	O
some	NN	O	O
,	NN	O	O
if	NN	O	O
not	NN	O	O
all	NN	O	O
,	NN	O	O
adenovirus	NN	O	O
types	NN	O	O
during	NN	O	O
many	NN	O	O
years	NN	O	O
of	NN	O	O
symbiotic	NN	O	O
,	NN	O	O
persistent	NN	O	O
relationship	NN	O	O
in	NN	O	O
human	NN	O	O
tonsils	NN	O	O
and	NN	O	O
adenoids	NN	O	O
and	NN	O	O
possibly	NN	O	O
other	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
.	NN	O	O

-DOCSTART-	O

Preferential	NN	O	O
presentation	NN	O	O
of	NN	O	O
herpes	NN	O	B-protein
simplex	NN	O	I-protein
virus	NN	O	I-protein
T-cell	NN	O	I-protein
antigen	NN	O	I-protein
by	NN	O	O
HLA	NN	O	B-protein
DQA1*0501/DQB1*0201	NN	O	I-protein
in	NN	O	O
comparison	NN	O	O
to	NN	O	O
HLA	NN	O	B-protein
DQA1*0201/DQB1*0201	NN	O	I-protein
.	NN	O	O

The	NN	O	O
HLA	NN	O	B-DNA
DQA1	NN	O	I-DNA
locus	NN	O	I-DNA
is	NN	O	O
polymorphic	NN	O	O
.	NN	O	O

Haplotypes	NN	O	O
containing	NN	O	O
HLA	NN	O	B-DNA
DQA1*0501	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
HLA	NN	O	B-DNA
DQA1*0201	NN	O	I-DNA
,	NN	O	O
together	NN	O	O
with	NN	O	O
HLA	NN	O	B-DNA
DQB1*0201	NN	O	I-DNA
are	NN	O	O
associated	NN	O	O
with	NN	O	O
Grave	NN	O	O
's	NN	O	O
disease	NN	O	O
and	NN	O	O
celiac	NN	O	O
sprue	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
functional	NN	O	O
correlate	NN	O	O
of	NN	O	O
DQA1	NN	O	O
polymorphism	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
infiltrating	NN	O	O
a	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
virus	NN	O	O
(	NN	O	O
HSV	NN	O	O
)	NN	O	O
lesion	NN	O	O
from	NN	O	O
a	NN	O	O
HLA	NN	O	O
DQ	NN	O	O
2	NN	O	O
,	NN	O	O
7	NN	O	O
individual	NN	O	O
yielded	NN	O	O
a	NN	O	O
virus-specific	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
clone	NN	O	I-cell_line
restricted	NN	O	O
by	NN	O	O
DQ2	NN	O	B-protein
.	NN	O	O

Presentation	NN	O	O
of	NN	O	O
viral	NN	O	O
peptide	NN	O	O
and	NN	O	O
protein	NN	O	O
segregated	NN	O	O
with	NN	O	O
DQA1	NN	O	B-DNA
allele	NN	O	I-DNA
,	NN	O	O
because	NN	O	O
cell	NN	O	O
lines	NN	O	O
bearing	NN	O	O
DQA1*0501/DQB1*0201	NN	O	B-protein
heterodimers	NN	O	I-protein
presented	NN	O	O
antigen	NN	O	O
in	NN	O	O
proliferation	NN	O	O
and	NN	O	O
cytotoxicity	NN	O	O
assays	NN	O	O
much	NN	O	O
more	NN	O	O
efficiently	NN	O	O
than	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
bearing	NN	O	O
DQA1*0201/DQB1*0201	NN	O	B-protein
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
viral	NN	O	O
peptide	NN	O	O
to	NN	O	O
cell	NN	O	O
lines	NN	O	O
bearing	NN	O	O
DQA1*0201	NN	O	B-protein
,	NN	O	O
in	NN	O	O
comparison	NN	O	O
to	NN	O	O
DQA1*0501	NN	O	B-protein
,	NN	O	O
was	NN	O	O
only	NN	O	O
moderately	NN	O	O
reduced	NN	O	O
and	NN	O	O
may	NN	O	O
not	NN	O	O
explain	NN	O	O
this	NN	O	O
effect	NN	O	O
.	NN	O	O

Truncation	NN	O	O
and	NN	O	O
substitution	NN	O	O
analyses	NN	O	O
of	NN	O	O
peptide	NN	O	O
binding	NN	O	O
and	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
were	NN	O	O
performed	NN	O	O
to	NN	O	O
determine	NN	O	O
which	NN	O	O
viral	NN	O	O
peptide	NN	O	O
residues	NN	O	O
contacting	NN	O	O
TCR	NN	O	B-protein
might	NN	O	O
therefore	NN	O	O
be	NN	O	O
presented	NN	O	O
in	NN	O	O
an	NN	O	O
altered	NN	O	O
conformation	NN	O	O
by	NN	O	O
DQA1*0201/DQB1*0201	NN	O	B-protein
.	NN	O	O

Residues	NN	O	O
432	NN	O	O
,	NN	O	O
435	NN	O	O
,	NN	O	O
437	NN	O	O
,	NN	O	O
438	NN	O	O
,	NN	O	O
and	NN	O	O
440	NN	O	O
(	NN	O	O
position	NN	O	B-DNA
P1	NN	O	I-DNA
,	NN	O	I-DNA
P4	NN	O	I-DNA
,	NN	O	I-DNA
P6	NN	O	I-DNA
,	NN	O	I-DNA
P7	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
P9	NN	O	I-DNA
)	NN	O	O
contributed	NN	O	O
to	NN	O	O
DQ2	NN	O	B-protein
binding	NN	O	O
,	NN	O	O
whereas	NN	O	O
residues	NN	O	O
431	NN	O	O
,	NN	O	O
433	NN	O	O
,	NN	O	O
434	NN	O	O
,	NN	O	O
and	NN	O	O
436	NN	O	O
(	NN	O	O
positions	NN	O	B-DNA
P	NN	O	I-DNA
1	NN	O	I-DNA
,	NN	O	I-DNA
P2	NN	O	I-DNA
,	NN	O	I-DNA
P3	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
P5	NN	O	I-DNA
)	NN	O	O
contributed	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
contact	NN	O	O
.	NN	O	O

Differential	NN	O	O
presentation	NN	O	O
of	NN	O	O
peptide	NN	O	O
by	NN	O	O
HLA	NN	O	B-protein
DQ2	NN	O	I-protein
heterodimers	NN	O	I-protein
varying	NN	O	O
at	NN	O	O
the	NN	O	O
DQA1	NN	O	B-DNA
locus	NN	O	I-DNA
may	NN	O	O
have	NN	O	O
relevance	NN	O	O
to	NN	O	O
host	NN	O	O
defense	NN	O	O
and	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
HLA	NN	O	O
DQ2	NN	O	B-protein
-associated	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibitor	NN	O	O
(	NN	O	O
IK	NN	O	O
)	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
induced	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
expression	NN	O	O
also	NN	O	O
inhibits	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
constitutive	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
Raji	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	I-protein
MHC	NN	O	I-protein
)	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
is	NN	O	O
constitutive	NN	O	O
in	NN	O	O
professional	NN	O	B-cell_type
antigen	NN	O	I-cell_type
presenting	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
APCs	NN	O	B-cell_type
)	NN	O	O
but	NN	O	O
can	NN	O	O
also	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
non	NN	O	O
professional	NN	O	B-cell_type
APCs	NN	O	I-cell_type
(	NN	O	O
e.g	NN	O	O
.	NN	O	O
fibroblasts	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
characterised	NN	O	O
a	NN	O	O
new	NN	O	O
factor	NN	O	O
called	NN	O	O
IK	NN	O	B-protein
which	NN	O	O
is	NN	O	O
an	NN	O	O
efficient	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
novel	NN	O	O
role	NN	O	O
for	NN	O	O
IK	NN	O	B-protein
in	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
since	NN	O	O
over-expression	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
by	NN	O	O
stable	NN	O	O
transfection	NN	O	O
into	NN	O	O
human	NN	O	O
B	NN	O	O
cells	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
total	NN	O	O
disappearance	NN	O	O
of	NN	O	O
constitutive	NN	O	O
MHC	NN	O	B-RNA
class	NN	O	I-RNA
II	NN	O	I-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

The	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
transactivator	NN	O	I-DNA
(	NN	O	O
CIITA	NN	O	B-DNA
)	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
both	NN	O	O
constitutive	NN	O	O
and	NN	O	O
IFN-gamma	NN	O	B-protein
induced	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expressions	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
CIITA	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
IK	NN	O	B-protein
stably	NN	O	O
transfected	NN	O	O
clones	NN	O	O
revealed	NN	O	O
a	NN	O	O
marked	NN	O	O
reduction	NN	O	O
of	NN	O	O
CIITA	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcription	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
these	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
IK	NN	O	B-protein
protein	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
and	NN	O	O
that	NN	O	O
inhibition	NN	O	O
induced	NN	O	O
by	NN	O	O
IK	NN	O	B-protein
is	NN	O	O
upstream	NN	O	O
of	NN	O	O
CIITA	NN	O	B-DNA
in	NN	O	O
this	NN	O	O
regulatory	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Selective	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
interleukin-12	NN	O	B-protein
receptor	NN	O	I-protein
component	NN	O	I-protein
by	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
1	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Interleukin-12	NN	O	B-protein
(	NN	O	O
IL-12	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
heterodimeric	NN	O	B-protein
cytokine	NN	O	I-protein
produced	NN	O	O
by	NN	O	O
activated	NN	O	O
monocytes	NN	O	O
and	NN	O	O
dendritic	NN	O	O
cells	NN	O	O
,	NN	O	O
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
interferon	NN	O	B-protein
(	NN	O	I-protein
IFN	NN	O	I-protein
)	NN	O	I-protein
-gamma	NN	O	I-protein
production	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
IFN-gamma-producing	NN	O	B-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
1	NN	O	I-cell_type
(	NN	O	I-cell_type
Th1	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
IL-12	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
IL-12R	NN	O	I-protein
)	NN	O	I-protein
beta	NN	O	I-protein
2	NN	O	I-protein
subunit	NN	O	I-protein
,	NN	O	O
a	NN	O	O
recently	NN	O	O
cloned	NN	O	O
binding	NN	O	B-protein
and	NN	O	I-protein
signal	NN	O	I-protein
transducing	NN	O	I-protein
component	NN	O	I-protein
of	NN	O	O
the	NN	O	O
IL-12R	NN	O	B-protein
,	NN	O	O
is	NN	O	O
expressed	NN	O	O
on	NN	O	O
human	NN	O	B-cell_line
Th1	NN	O	I-cell_line
but	NN	O	I-cell_line
not	NN	O	I-cell_line
Th2	NN	O	I-cell_line
clones	NN	O	I-cell_line
and	NN	O	O
is	NN	O	O
induced	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
naive	NN	O	I-cell_type
cells	NN	O	I-cell_type
along	NN	O	O
the	NN	O	O
Th1	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
Th2	NN	O	O
pathway	NN	O	O
.	NN	O	O

IL-12	NN	O	B-protein
and	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
but	NN	O	I-protein
not	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
interferons	NN	O	I-protein
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-12R	NN	O	B-protein
beta	NN	O	I-protein
2	NN	O	I-protein
chain	NN	O	I-protein
during	NN	O	O
in	NN	O	O
vitro	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
after	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
triggering	NN	O	O
.	NN	O	O

The	NN	O	O
selective	NN	O	O
expression	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-12R	NN	O	B-protein
beta	NN	O	I-protein
2	NN	O	I-protein
subunit	NN	O	I-protein
may	NN	O	O
help	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
Th1/Th2	NN	O	O
differentiation	NN	O	O
and	NN	O	O
may	NN	O	O
provide	NN	O	O
therapeutic	NN	O	O
options	NN	O	O
for	NN	O	O
altering	NN	O	O
the	NN	O	O
Th1/Th2	NN	O	O
balance	NN	O	O
in	NN	O	O
several	NN	O	O
immuno-pathological	NN	O	O
conditions	NN	O	O
such	NN	O	O
as	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
and	NN	O	O
allergies	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
activation	NN	O	I-protein
factor	NN	O	I-protein
NF-ATc	NN	O	B-protein
positively	NN	O	O
regulates	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
and	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Clinical	NN	O	O
deterioration	NN	O	O
in	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
infection	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
viral	NN	O	O
replication	NN	O	O
and	NN	O	O
burden	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
and	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
.	NN	O	O

T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
ensuing	NN	O	O
cellular	NN	O	O
gene	NN	O	O
activation	NN	O	O
can	NN	O	O
be	NN	O	O
critical	NN	O	O
for	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

The	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
may	NN	O	O
influence	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
is	NN	O	O
therefore	NN	O	O
compelling	NN	O	O
given	NN	O	O
the	NN	O	O
tight	NN	O	O
correlation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
to	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
certain	NN	O	O
NF-AT	NN	O	B-protein
(	NN	O	I-protein
Rel	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
productively	NN	O	O
bind	NN	O	O
the	NN	O	O
kappaB	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
synergize	NN	O	O
with	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
Tat	NN	O	B-protein
in	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
and	NN	O	O
enhance	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
link	NN	O	O
regulatory	NN	O	B-protein
factors	NN	O	I-protein
critical	NN	O	O
to	NN	O	O
T	NN	O	O
cell	NN	O	O
commitment	NN	O	O
directly	NN	O	O
to	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	O
mediated	NN	O	O
by	NN	O	O
NFAT	NN	O	B-protein
is	NN	O	O
highly	NN	O	O
inducible	NN	O	O
in	NN	O	O
effector	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
2	NN	O	I-cell_type
(	NN	O	I-cell_type
Th2	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Transcriptional	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
genes	NN	O	O
for	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
among	NN	O	O
others	NN	O	O
.	NN	O	O

Upon	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
,	NN	O	O
precursor	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
(	NN	O	I-cell_type
pTh	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
proliferate	NN	O	O
and	NN	O	O
differentiate	NN	O	O
into	NN	O	O
two	NN	O	O
populations	NN	O	O
of	NN	O	O
effector	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
eTh1	NN	O	B-cell_type
and	NN	O	O
eTh2	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
each	NN	O	O
one	NN	O	O
expressing	NN	O	O
a	NN	O	O
specific	NN	O	O
pattern	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
distinguishes	NN	O	O
them	NN	O	O
from	NN	O	O
their	NN	O	O
precursors	NN	O	O
.	NN	O	O

eTh2	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
the	NN	O	O
major	NN	O	O
source	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
while	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
is	NN	O	O
produced	NN	O	O
by	NN	O	O
eTh1	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
have	NN	O	O
used	NN	O	O
reporter	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
to	NN	O	O
show	NN	O	O
that	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
are	NN	O	O
transiently	NN	O	O
induced	NN	O	O
during	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
pTh	NN	O	B-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
either	NN	O	O
eTh1	NN	O	B-cell_type
or	NN	O	O
eTh2	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
as	NN	O	O
a	NN	O	O
common	NN	O	O
growth	NN	O	O
factor	NN	O	O
in	NN	O	O
both	NN	O	O
pathways	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
although	NN	O	O
NFAT	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
is	NN	O	O
similarly	NN	O	O
induced	NN	O	O
in	NN	O	O
both	NN	O	O
eTh1	NN	O	B-cell_type
and	NN	O	O
eTh2	NN	O	B-cell_type
cells	NN	O	I-cell_type
upon	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
,	NN	O	O
only	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
complexes	NN	O	I-protein
present	NN	O	O
in	NN	O	O
eTh2	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
able	NN	O	O
to	NN	O	O
mediate	NN	O	O
high-level	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
relatively	NN	O	O
little	NN	O	O
NFAT	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
was	NN	O	O
induced	NN	O	O
in	NN	O	O
eTh1	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
activated	NN	O	O
pTh	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
neither	NN	O	O
eTh1	NN	O	B-cell_type
nor	NN	O	O
eTh2	NN	O	B-cell_type
cells	NN	O	I-cell_type
produced	NN	O	O
significant	NN	O	O
IL-2	NN	O	B-protein
upon	NN	O	O
stimulation	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
transcriptional	NN	O	O
activities	NN	O	O
directly	NN	O	O
correlate	NN	O	O
with	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
production	NN	O	O
induced	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
in	NN	O	O
eTh2	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
activated	NN	O	O
NFAT	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
effector	NN	O	O
function	NN	O	O
of	NN	O	O
eTh2	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
the	NN	O	O
failure	NN	O	O
of	NN	O	O
eTh1	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
produce	NN	O	O
IL-4	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
an	NN	O	O
antigen	NN	O	O
is	NN	O	O
due	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
partially	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
failure	NN	O	O
to	NN	O	O
induce	NN	O	O
high-level	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

Regulation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
could	NN	O	O
be	NN	O	O
therefore	NN	O	O
a	NN	O	O
critical	NN	O	O
element	NN	O	O
in	NN	O	O
the	NN	O	O
polarization	NN	O	O
to	NN	O	O
eTh1	NN	O	B-cell_type
or	NN	O	O
eTh2	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
in	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

During	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
serious	NN	O	O
bacterial	NN	O	O
infections	NN	O	O
,	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
interacts	NN	O	O
with	NN	O	O
monocyte/macrophage	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
is	NN	O	O
crucial	NN	O	O
in	NN	O	O
activating	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
encoding	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
by	NN	O	O
LPS	NN	O	O
in	NN	O	O
a	NN	O	O
promonocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
U937	NN	O	B-cell_line
)	NN	O	O
followed	NN	O	O
a	NN	O	O
rather	NN	O	O
slow	NN	O	O
kinetics	NN	O	O
,	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
inhibitor	NN	O	O
hydrolysis	NN	O	O
.	NN	O	O

No	NN	O	O
degradation	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
and	NN	O	I-protein
p100	NN	O	I-protein
inhibitors	NN	O	I-protein
was	NN	O	O
observed	NN	O	O
under	NN	O	O
these	NN	O	O
conditions	NN	O	O
.	NN	O	O

The	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
leading	NN	O	O
to	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
involved	NN	O	O
the	NN	O	O
intracellular	NN	O	O
generation	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
,	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
the	NN	O	O
concomitant	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
antioxidants	NN	O	O
on	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
the	NN	O	O
emission	NN	O	O
of	NN	O	O
a	NN	O	O
fluorescent	NN	O	O
probe	NN	O	O
reacting	NN	O	O
intracellularly	NN	O	O
with	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
.	NN	O	O

This	NN	O	O
ROS	NN	O	O
pathway	NN	O	O
was	NN	O	O
also	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
other	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
indicates	NN	O	O
that	NN	O	O
phospholipase	NN	O	B-protein
A2	NN	O	I-protein
and	NN	O	O
5-lipoxygenase	NN	O	B-protein
are	NN	O	O
also	NN	O	O
involved	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
pathway	NN	O	O
involving	NN	O	O
the	NN	O	O
acidic	NN	O	B-protein
sphingomyelinase	NN	O	I-protein
of	NN	O	O
the	NN	O	O
endolysosomial	NN	O	O
membrane	NN	O	O
did	NN	O	O
not	NN	O	O
seem	NN	O	O
to	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
LPS-induced	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

c-Rel	NN	O	B-protein
is	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
pentoxifylline-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
possible	NN	O	O
clinical	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
methyl	NN	O	O
xanthine	NN	O	O
derivative	NN	O	O
,	NN	O	O
pentoxifylline	NN	O	O
(	NN	O	O
PF	NN	O	O
)	NN	O	O
,	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
T	NN	O	O
cell-dependent	NN	O	O
diseases	NN	O	O
is	NN	O	O
being	NN	O	O
noted	NN	O	O
with	NN	O	O
increasing	NN	O	O
interest	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
molecular	NN	O	O
consequences	NN	O	O
of	NN	O	O
PF	NN	O	O
treatment	NN	O	O
during	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
anti-CD3-induced	NN	O	O
c-Rel	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
PF	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
other	NN	O	O
NF-kappaB	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
was	NN	O	O
not	NN	O	O
significantly	NN	O	O
affected	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
which	NN	O	O
has	NN	O	O
the	NN	O	O
same	NN	O	O
signaling	NN	O	O
requirements	NN	O	O
as	NN	O	O
c-Rel	NN	O	B-protein
induction	NN	O	O
,	NN	O	O
was	NN	O	O
not	NN	O	O
inhibited	NN	O	O
by	NN	O	O
PF	NN	O	O
.	NN	O	O

Among	NN	O	O
genes	NN	O	O
that	NN	O	O
respond	NN	O	O
to	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
was	NN	O	O
suppressed	NN	O	O
by	NN	O	O
PF	NN	O	O
,	NN	O	O
whereas	NN	O	O
IL-2R	NN	O	B-RNA
(	NN	O	I-RNA
alpha	NN	O	I-RNA
)	NN	O	I-RNA
chain	NN	O	I-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
was	NN	O	O
not	NN	O	O
affected	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
implicated	NN	O	O
c-Rel	NN	O	B-protein
as	NN	O	O
an	NN	O	O
IL-2	NN	O	B-protein
promoter	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
for	NN	O	O
which	NN	O	O
experimental	NN	O	O
support	NN	O	O
was	NN	O	O
obtained	NN	O	O
from	NN	O	O
transient	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
with	NN	O	O
the	NN	O	O
observation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
c-Rel	NN	O	B-protein
induction	NN	O	O
was	NN	O	O
not	NN	O	O
blocked	NN	O	O
by	NN	O	O
PF	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
greater	NN	O	O
selectivity	NN	O	O
of	NN	O	O
PF	NN	O	O
,	NN	O	O
compared	NN	O	O
with	NN	O	O
FK506	NN	O	O
,	NN	O	O
at	NN	O	O
both	NN	O	O
the	NN	O	O
molecular	NN	O	O
and	NN	O	O
cellular	NN	O	O
levels	NN	O	O
may	NN	O	O
prove	NN	O	O
advantageous	NN	O	O
in	NN	O	O
manipulating	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

V3	NN	O	B-protein
loop	NN	O	I-protein
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
suppresses	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
-induced	NN	O	O
T	NN	O	O
cell	NN	O	O
growth	NN	O	O
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
AIDS	NN	O	O
Res	NN	O	O
Hum	NN	O	O
Retroviruses	NN	O	O
1997	NN	O	O
May	NN	O	O
1	NN	O	O
;	NN	O	O
13	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
:	NN	O	O
633	NN	O	O
]	NN	O	O

We	NN	O	O
tested	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
three	NN	O	O
linear	NN	O	O
or	NN	O	O
two	NN	O	O
loop	NN	O	O
peptides	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
V3	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HTLV-III	NN	O	O
BH10	NN	O	O
clone	NN	O	O
or	NN	O	O
the	NN	O	O
SF2	NN	O	O
strain	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
on	NN	O	O
IL-2	NN	O	B-protein
-driven	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

V3-BH10	NN	O	B-protein
,	NN	O	O
which	NN	O	O
consists	NN	O	O
of	NN	O	O
42	NN	O	B-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
and	NN	O	O
has	NN	O	O
a	NN	O	O
loop	NN	O	B-protein
structure	NN	O	I-protein
,	NN	O	O
suppressed	NN	O	O
IL-2	NN	O	B-protein
-driven	NN	O	O
proliferation	NN	O	O
of	NN	O	O
all	NN	O	O
IL-2-dependent	NN	O	B-cell_line
cells	NN	O	I-cell_line
[	NN	O	O
Kit225	NN	O	B-cell_line
,	NN	O	O
ED-40515	NN	O	B-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
,	NN	O	O
KT-3	NN	O	B-cell_line
,	NN	O	O
7-day	NN	O	B-cell_line
PHA-blasts	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
fresh	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
]	NN	O	O
tested	NN	O	O
,	NN	O	O
whereas	NN	O	O
it	NN	O	O
did	NN	O	O
not	NN	O	O
suppress	NN	O	O
the	NN	O	O
cell	NN	O	O
growth	NN	O	O
of	NN	O	O
IL-2-independent	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
Hut102	NN	O	B-cell_line
,	NN	O	O
Molt-4	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
Jurkat	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

This	NN	O	O
suppressive	NN	O	O
effect	NN	O	O
was	NN	O	O
also	NN	O	O
seen	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
-driven	NN	O	O
cell	NN	O	O
growth	NN	O	O
of	NN	O	O
CD8-positive	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
purified	NN	O	O
from	NN	O	O
7-day	NN	O	B-cell_type
PHA-blasts	NN	O	I-cell_type
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
CD4	NN	O	B-protein
molecules	NN	O	I-protein
were	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
suppression	NN	O	O
.	NN	O	O

The	NN	O	O
treatment	NN	O	O
with	NN	O	O
anti-V3	NN	O	B-protein
loop	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
902	NN	O	B-protein
antibody	NN	O	I-protein
)	NN	O	O
completely	NN	O	O
abolished	NN	O	O
the	NN	O	O
suppressive	NN	O	O
effect	NN	O	O
of	NN	O	O
V3-BH10	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
V3-BH10	NN	O	B-protein
generated	NN	O	O
the	NN	O	O
arrest	NN	O	O
of	NN	O	O
Kit225	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
also	NN	O	O
purified	NN	O	B-cell_line
CD8-positive	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
in	NN	O	O
G1	NN	O	O
phase	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Neither	NN	O	O
chromatin	NN	O	B-DNA
condensation	NN	O	O
nor	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
Kit225	NN	O	B-cell_line
cells	NN	O	I-cell_line
cultured	NN	O	O
with	NN	O	O
V3-BH10	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

V3-BH10	NN	O	B-protein
neither	NN	O	O
blocked	NN	O	O
radiolabeled	NN	O	O
IL-2	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
receptors	NN	O	I-protein
nor	NN	O	O
affected	NN	O	O
tyrosyl	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
several	NN	O	O
cellular	NN	O	B-protein
proteins	NN	O	I-protein
(	NN	O	O
p120	NN	O	B-protein
,	NN	O	O
p98	NN	O	B-protein
,	NN	O	O
p96	NN	O	B-protein
,	NN	O	O
p54	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p38	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
immediately	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
V3-BH10	NN	O	B-protein
enhanced	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
but	NN	O	O
not	NN	O	O
c-myc	NN	O	B-DNA
or	NN	O	O
junB	NN	O	B-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
V3	NN	O	B-protein
loop	NN	O	I-protein
of	NN	O	O
gp120	NN	O	B-protein
to	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
molecule	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
appears	NN	O	O
to	NN	O	O
affect	NN	O	O
intracellular	NN	O	O
IL-2	NN	O	B-protein
signaling	NN	O	O
,	NN	O	O
which	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
T	NN	O	O
cell	NN	O	O
growth	NN	O	O

-DOCSTART-	O

A	NN	O	O
negative	NN	O	O
role	NN	O	O
for	NN	O	O
phosphoinositide	NN	O	B-protein
3-kinase	NN	O	I-protein
in	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
function	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
A	NN	O	O
delicate	NN	O	O
balance	NN	O	O
between	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
determines	NN	O	O
the	NN	O	O
specificity	NN	O	O
and	NN	O	O
magnitude	NN	O	O
of	NN	O	O
an	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Phosphoinositide	NN	O	B-protein
3-kinase	NN	O	I-protein
(	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
)	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
a	NN	O	O
diverse	NN	O	O
set	NN	O	O
of	NN	O	O
receptors	NN	O	O
that	NN	O	O
determine	NN	O	O
T-cell	NN	O	O
function	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
costimulatory	NN	O	B-protein
receptor	NN	O	I-protein
CD28	NN	O	I-protein
,	NN	O	O
and	NN	O	O
negative	NN	O	B-protein
regulators	NN	O	I-protein
of	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
such	NN	O	O
as	NN	O	O
CTLA-4	NN	O	B-protein
.	NN	O	O

PI	NN	O	B-protein
3-kinase	NN	O	I-protein
is	NN	O	O
also	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
haematopoietic	NN	O	O
cytokines	NN	O	O
that	NN	O	O
determine	NN	O	O
T-cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
lymphocyte	NN	O	B-cell_type
proliferation	NN	O	O
.	NN	O	O

PI	NN	O	B-protein
3-kinase	NN	O	I-protein
can	NN	O	O
thus	NN	O	O
dynamically	NN	O	O
influence	NN	O	O
the	NN	O	O
outcome	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
reactions	NN	O	O
at	NN	O	O
various	NN	O	O
stages	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
in	NN	O	O
TCR	NN	O	B-protein
-directed	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
using	NN	O	O
activated	NN	O	O
or	NN	O	O
inhibitory	NN	O	O
versions	NN	O	O
of	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Certain	NN	O	O
aspects	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
responses	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
AP1	NN	O	B-protein
and	NN	O	O
serum	NN	O	B-protein
response	NN	O	I-protein
factor	NN	O	I-protein
were	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
.	NN	O	O

We	NN	O	O
found	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
that	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
profoundly	NN	O	O
influenced	NN	O	O
the	NN	O	O
transactivation	NN	O	O
capacity	NN	O	O
of	NN	O	O
'	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
'	NN	O	O
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
elicited	NN	O	O
by	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
:	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
activated	NN	O	O
form	NN	O	O
of	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
inhibited	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
NF-AT	NN	O	B-protein
responses	NN	O	O
,	NN	O	O
whereas	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
mutant	NN	O	O
of	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
potently	NN	O	O
enhanced	NN	O	O
TCR	NN	O	B-protein
-controlled	NN	O	O
NF-AT	NN	O	B-protein
induction	NN	O	O
.	NN	O	O

These	NN	O	O
effects	NN	O	O
of	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
were	NN	O	O
not	NN	O	O
mediated	NN	O	O
by	NN	O	O
previously	NN	O	O
identified	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
effectors	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
positive	NN	O	B-protein
regulator	NN	O	I-protein
of	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
,	NN	O	O
or	NN	O	O
the	NN	O	O
GTPase	NN	O	B-protein
Rac	NN	O	I-protein
,	NN	O	O
and	NN	O	O
are	NN	O	O
therefore	NN	O	O
likely	NN	O	O
to	NN	O	O
involve	NN	O	O
a	NN	O	O
novel	NN	O	O
,	NN	O	O
as	NN	O	O
yet	NN	O	O
unknown	NN	O	O
,	NN	O	O
effector	NN	O	B-protein
molecule	NN	O	I-protein
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Our	NN	O	O
results	NN	O	O
establish	NN	O	O
that	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
can	NN	O	O
both	NN	O	O
positively	NN	O	O
and	NN	O	O
negatively	NN	O	O
regulate	NN	O	O
T-cell	NN	O	O
function	NN	O	O
,	NN	O	O
and	NN	O	O
uncover	NN	O	O
a	NN	O	O
previously	NN	O	O
unrecognized	NN	O	O
function	NN	O	O
for	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
a	NN	O	O
selective	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
TCR	NN	O	B-protein
-signalling	NN	O	O
events	NN	O	O
and	NN	O	O
therefore	NN	O	O
as	NN	O	O
a	NN	O	O
determinant	NN	O	O
of	NN	O	O
T-cell	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Pax-5	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
during	NN	O	O
B-cell	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Pax-5	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
during	NN	O	O
the	NN	O	O
early	NN	O	O
stages	NN	O	O
of	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
influences	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
B-cell-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
existing	NN	O	O
isoform	NN	O	O
(	NN	O	O
Pax-5	NN	O	B-protein
,	NN	O	O
which	NN	O	O
we	NN	O	O
have	NN	O	O
named	NN	O	O
Pax-5a	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
isolated	NN	O	O
three	NN	O	O
new	NN	O	O
isoforms	NN	O	O
,	NN	O	O
Pax-5b	NN	O	B-protein
,	NN	O	O
Pax-5d	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Pax-5e	NN	O	B-protein
,	NN	O	O
from	NN	O	O
murine	NN	O	O
spleen	NN	O	O
and	NN	O	O
B-lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
using	NN	O	O
library	NN	O	O
screenings	NN	O	O
and	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
amplification	NN	O	O
.	NN	O	O

Isoforms	NN	O	O
Pax-5b	NN	O	B-protein
and	NN	O	O
Pax-5e	NN	O	B-protein
have	NN	O	O
spliced	NN	O	O
out	NN	O	O
their	NN	O	O
second	NN	O	O
exon	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
proteins	NN	O	O
with	NN	O	O
only	NN	O	O
a	NN	O	O
partial	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

Isoforms	NN	O	O
Pax-5d	NN	O	O
and	NN	O	O
Pax-5e	NN	O	B-protein
have	NN	O	O
deleted	NN	O	O
the	NN	O	O
3'-region	NN	O	B-protein
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
the	NN	O	O
transactivating	NN	O	O
domain	NN	O	O
,	NN	O	O
and	NN	O	O
replaced	NN	O	O
it	NN	O	O
with	NN	O	O
a	NN	O	O
novel	NN	O	O
sequence	NN	O	O
.	NN	O	O

The	NN	O	O
existence	NN	O	O
of	NN	O	O
alternative	NN	O	O
Pax-5	NN	O	B-RNA
transcripts	NN	O	I-RNA
was	NN	O	O
confirmed	NN	O	O
using	NN	O	O
RNase	NN	O	B-protein
protection	NN	O	O
assays	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
Pax-5a	NN	O	B-protein
and	NN	O	O
Pax-5b	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
using	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

Pax-5a	NN	O	B-protein
was	NN	O	O
detectable	NN	O	O
in	NN	O	O
pro-	NN	O	B-cell_line
,	NN	O	I-cell_line
pre-	NN	O	I-cell_line
,	NN	O	I-cell_line
and	NN	O	I-cell_line
mature	NN	O	I-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
two	NN	O	O
plasmacytomas	NN	O	O
;	NN	O	O
Pax-5b	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
present	NN	O	O
at	NN	O	O
low	NN	O	O
levels	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
,	NN	O	O
unexpectedly	NN	O	O
,	NN	O	O
in	NN	O	O
one	NN	O	O
plasma	NN	O	O
cell	NN	O	O
line	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
pro-B-cell	NN	O	B-cell_line
or	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
in	NN	O	O
vitro	NN	O	O
translated	NN	O	O
Pax-5a	NN	O	B-protein
and	NN	O	O
Pax-5d	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Pax-5b	NN	O	B-protein
or	NN	O	O
Pax-5e	NN	O	B-protein
,	NN	O	O
could	NN	O	O
interact	NN	O	O
with	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	B-DNA
activator	NN	O	I-DNA
protein-binding	NN	O	I-DNA
site	NN	O	I-DNA
on	NN	O	O
the	NN	O	O
blk	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
this	NN	O	O
assay	NN	O	O
,	NN	O	O
we	NN	O	O
also	NN	O	O
showed	NN	O	O
that	NN	O	O
Pax-5d	NN	O	B-protein
was	NN	O	O
present	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
some	NN	O	O
(	NN	O	O
but	NN	O	O
not	NN	O	O
all	NN	O	O
)	NN	O	O
B-lymphoid	NN	O	B-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	B-DNA
activator	NN	O	I-DNA
protein-binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
pattern	NN	O	O
of	NN	O	O
differential	NN	O	O
expression	NN	O	O
of	NN	O	O
alternatively	NN	O	B-protein
spliced	NN	O	I-protein
Pax-5	NN	O	I-protein
isoforms	NN	O	I-protein
suggests	NN	O	O
that	NN	O	O
they	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
regulators	NN	O	B-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
during	NN	O	O
B-cell	NN	O	O
maturation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reconstitution	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
antigen	NN	O	O
receptor-induced	NN	O	O
Erk2	NN	O	B-protein
kinase	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
Lck-negative	NN	O	B-cell_line
JCaM1	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
Syk	NN	O	B-protein
.	NN	O	O

The	NN	O	O
two	NN	O	O
related	NN	O	O
protein-tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
Syk	NN	O	B-protein
and	NN	O	O
Zap	NN	O	B-protein
are	NN	O	O
rapidly	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
residues	NN	O	O
and	NN	O	O
enzymatically	NN	O	O
activated	NN	O	O
upon	NN	O	O
crosslinking	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Syk	NN	O	B-protein
is	NN	O	O
less	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
Src	NN	O	B-protein
family	NN	O	I-protein
kinase	NN	O	I-protein
Lck	NN	O	B-protein
than	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Zap	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
Syk	NN	O	B-protein
in	NN	O	O
the	NN	O	O
Lck-negative	NN	O	B-cell_line
JCaM1	NN	O	I-cell_line
cells	NN	O	I-cell_line
enabled	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor/CD3	NN	O	I-protein
complex	NN	O	I-protein
to	NN	O	O
induce	NN	O	O
a	NN	O	O
normal	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
pathway	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
reporter	NN	O	O
construct	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
Zap	NN	O	B-protein
and	NN	O	O
other	NN	O	O
protein-tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
were	NN	O	O
unable	NN	O	O
to	NN	O	O
reconstitute	NN	O	O
these	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
when	NN	O	O
expressed	NN	O	O
at	NN	O	O
the	NN	O	O
same	NN	O	O
levels	NN	O	O
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
,	NN	O	O
Syk	NN	O	B-protein
was	NN	O	O
phosphorylated	NN	O	O
on	NN	O	O
tyrosine	NN	O	O
,	NN	O	O
while	NN	O	O
Zap	NN	O	B-protein
was	NN	O	O
not	NN	O	O
.	NN	O	O

The	NN	O	O
Syk	NN	O	B-protein
-mediated	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
-induced	NN	O	O
MAPK	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
detectable	NN	O	O
within	NN	O	O
1	NN	O	O
min	NN	O	O
of	NN	O	O
receptor	NN	O	O
stimulation	NN	O	O
and	NN	O	O
peaked	NN	O	O
at	NN	O	O
3-5	NN	O	O
min	NN	O	O
.	NN	O	O

The	NN	O	O
capacity	NN	O	O
of	NN	O	O
Syk	NN	O	B-protein
to	NN	O	O
reconstitute	NN	O	O
the	NN	O	O
MAPK	NN	O	B-protein
response	NN	O	O
required	NN	O	O
the	NN	O	O
catalytic	NN	O	O
activity	NN	O	O
of	NN	O	O
Syk	NN	O	B-protein
,	NN	O	O
an	NN	O	O
intact	NN	O	O
autophosphorylation	NN	O	B-protein
site	NN	O	I-protein
(	NN	O	O
Y518	NN	O	O
and	NN	O	O
Y519	NN	O	O
)	NN	O	O
,	NN	O	O
both	NN	O	O
Src	NN	O	B-protein
homology	NN	O	I-protein
2	NN	O	I-protein
domains	NN	O	I-protein
and	NN	O	O
it	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
inhibitory	NN	O	B-protein
N17-mutated	NN	O	I-protein
dominant-negative	NN	O	I-protein
Ras	NN	O	I-protein
construct	NN	O	I-protein
.	NN	O	O

A	NN	O	O
Y341	NN	O	O
--	NN	O	O
>	NN	O	O
F	NN	O	B-protein
mutant	NN	O	I-protein
of	NN	O	O
Syk	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
deficient	NN	O	O
in	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
phospholipase	NN	O	B-protein
Cy1	NN	O	I-protein
and	NN	O	O
Vav	NN	O	B-protein
,	NN	O	O
was	NN	O	O
less	NN	O	O
efficient	NN	O	O
than	NN	O	O
wild-type	NN	O	B-protein
Syk	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
Syk	NN	O	B-protein
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
Zap	NN	O	B-protein
,	NN	O	O
can	NN	O	O
transduce	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
independently	NN	O	O
of	NN	O	O
Lck	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
MEF2C	NN	O	B-protein
by	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
p38	NN	O	I-protein
in	NN	O	O
inflammation	NN	O	O
.	NN	O	O

For	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
innate	NN	O	O
immune	NN	O	O
system	NN	O	O
to	NN	O	O
mount	NN	O	O
a	NN	O	O
host	NN	O	O
defence	NN	O	O
response	NN	O	O
to	NN	O	O
infection	NN	O	O
,	NN	O	O
they	NN	O	O
must	NN	O	O
recognize	NN	O	O
products	NN	O	O
of	NN	O	O
microbial	NN	O	O
pathogens	NN	O	O
such	NN	O	O
as	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
endotoxin	NN	O	O
secreted	NN	O	O
by	NN	O	O
Gram-negative	NN	O	O
bacteria	NN	O	O
.	NN	O	O

These	NN	O	O
cellular	NN	O	O
responses	NN	O	O
require	NN	O	O
intracellular	NN	O	O
signalling	NN	O	O
pathways	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
four	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
pathways	NN	O	O
.	NN	O	O

In	NN	O	O
mammalian	NN	O	O
cells	NN	O	O
the	NN	O	O
MAPK	NN	O	B-protein
p38	NN	O	I-protein
is	NN	O	O
thought	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cellular	NN	O	O
responses	NN	O	O
during	NN	O	O
infection	NN	O	O
through	NN	O	O
its	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
proinflammatory	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

One	NN	O	O
means	NN	O	O
of	NN	O	O
understanding	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
in	NN	O	O
these	NN	O	O
responses	NN	O	O
is	NN	O	O
to	NN	O	O
identify	NN	O	O
proteins	NN	O	O
with	NN	O	O
functions	NN	O	O
regulated	NN	O	O
by	NN	O	O
p38	NN	O	B-protein
-catalysed	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
link	NN	O	O
between	NN	O	O
the	NN	O	O
p38	NN	O	B-protein
pathway	NN	O	O
and	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
myocyte-enhancer	NN	O	B-protein
factor	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	I-protein
MEF2	NN	O	I-protein
)	NN	O	I-protein
group	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
LPS	NN	O	O
increases	NN	O	O
the	NN	O	O
transactivation	NN	O	O
activity	NN	O	O
of	NN	O	O
MEF2C	NN	O	B-protein
through	NN	O	O
p38	NN	O	B-protein
-catalysed	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

One	NN	O	O
consequence	NN	O	O
of	NN	O	O
MEF2C	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
increased	NN	O	O
c-jun	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
p38	NN	O	B-protein
may	NN	O	O
influence	NN	O	O
host	NN	O	O
defence	NN	O	O
and	NN	O	O
inflammation	NN	O	O
by	NN	O	O
maintaining	NN	O	O
the	NN	O	O
balance	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
protein	NN	O	I-protein
consumed	NN	O	O
during	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cellular	NN	O	O
redox	NN	O	O
status	NN	O	O
influences	NN	O	O
both	NN	O	O
cytotoxic	NN	O	O
and	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
activation	NN	O	O
in	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
cellular	NN	O	O
redox	NN	O	O
status	NN	O	O
in	NN	O	O
both	NN	O	O
cytotoxic	NN	O	O
activity	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
investigated	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
either	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
from	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
PBL	NN	O	B-cell_type
)	NN	O	O
or	NN	O	O
long-term	NN	O	B-cell_line
cultured	NN	O	I-cell_line
NK	NN	O	I-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
with	NN	O	O
specific	NN	O	O
cell	NN	O	O
targets	NN	O	O
results	NN	O	O
in	NN	O	O
an	NN	O	O
increased	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
measured	NN	O	O
by	NN	O	O
gel	NN	O	O
retardation	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
pyrrolidine	NN	O	O
dithiocarbarmate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
but	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
was	NN	O	O
superinduced	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
PDTC	NN	O	O
paralleled	NN	O	O
with	NN	O	O
an	NN	O	O
inhibition	NN	O	O
of	NN	O	O
spontaneous	NN	O	O
cytotoxicity	NN	O	O
mediated	NN	O	O
by	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
serine	NN	O	B-protein
proteases	NN	O	I-protein
,	NN	O	O
N-alpha-tosyl-L-lysine	NN	O	O
chloromethyl	NN	O	O
ketone	NN	O	O
and	NN	O	O
N-alpha-tosyl-L-phenylalanine	NN	O	O
chloromethyl	NN	O	O
ketone	NN	O	O
,	NN	O	O
also	NN	O	O
blocked	NN	O	O
the	NN	O	O
cytolytic	NN	O	O
activity	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
against	NN	O	O
the	NN	O	O
sensitive	NN	O	O
target	NN	O	O
K562	NN	O	B-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
NK	NN	O	O
activity	NN	O	O
was	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
the	NN	O	O
effector	NN	O	O
cells	NN	O	O
with	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
inhibitor	NN	O	O
N-acetyl-leu-leu-norleucinal	NN	O	O
which	NN	O	O
selectively	NN	O	O
inhibits	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
involved	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
post-transcriptional	NN	O	O
events	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
reactive	NN	O	O
intermediates	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
molecular	NN	O	O
processes	NN	O	O
related	NN	O	O
with	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
a	NN	O	O
cytotoxic	NN	O	O
response	NN	O	O
by	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
predominant	NN	O	O
E2F	NN	O	B-protein
complex	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
primary	NN	O	I-cell_type
haemopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
AML	NN	O	B-cell_type
blasts	NN	O	I-cell_type
contains	NN	O	O
E2F-4	NN	O	B-protein
,	NN	O	O
DP-1	NN	O	B-protein
and	NN	O	O
p130	NN	O	B-protein
.	NN	O	O

The	NN	O	O
E2F	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
thought	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
.	NN	O	O

There	NN	O	O
is	NN	O	O
now	NN	O	O
also	NN	O	O
increasing	NN	O	O
evidence	NN	O	O
that	NN	O	O
some	NN	O	O
family	NN	O	O
members	NN	O	O
may	NN	O	O
act	NN	O	O
as	NN	O	O
oncogenes	NN	O	B-DNA
or	NN	O	O
tumour	NN	O	B-DNA
suppressor	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
characterization	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
primary	NN	O	I-cell_type
haemopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
acute	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
leukaemia	NN	O	I-cell_type
(	NN	O	I-cell_type
AML	NN	O	I-cell_type
)	NN	O	I-cell_type
blasts	NN	O	I-cell_type
may	NN	O	O
thus	NN	O	O
give	NN	O	O
an	NN	O	O
insight	NN	O	O
to	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
governing	NN	O	O
proliferation	NN	O	O
and	NN	O	O
leukaemogenesis	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Therefore	NN	O	O
we	NN	O	O
analysed	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
-DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
the	NN	O	O
constituent	NN	O	O
proteins	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
complexes	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
primary	NN	O	I-cell_type
haemopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
various	NN	O	O
lineages	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
studied	NN	O	O
blasts	NN	O	O
from	NN	O	O
18	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukaemia	NN	O	O
(	NN	O	O
AML	NN	O	O
)	NN	O	O
.	NN	O	O

On	NN	O	O
electromobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
a	NN	O	O
single	NN	O	O
E2F-DNA	NN	O	B-protein
binding	NN	O	I-protein
complex	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
which	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
contain	NN	O	O
E2F-4	NN	O	B-protein
,	NN	O	O
DP-1	NN	O	B-protein
and	NN	O	O
p130	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
all	NN	O	O
quiescent	NN	O	B-cell_type
haemopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
have	NN	O	O
the	NN	O	O
same	NN	O	O
complex	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
18	NN	O	O
AML	NN	O	O
samples	NN	O	O
by	NN	O	O
EMSA	NN	O	O
revealed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
binding	NN	O	O
and	NN	O	O
no	NN	O	O
gross	NN	O	O
abnormalities	NN	O	O
were	NN	O	O
detected	NN	O	O
.	NN	O	O

An	NN	O	O
E2F-4/p130	NN	O	B-protein
complex	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
representative	NN	O	O
samples	NN	O	O
of	NN	O	O
all	NN	O	O
FAB	NN	O	B-protein
types	NN	O	O
analysed	NN	O	O
.	NN	O	O

Thus	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
function	NN	O	O
are	NN	O	O
unlikely	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
primary	NN	O	O
pathogenic	NN	O	O
role	NN	O	O
in	NN	O	O
AML	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
a	NN	O	O
mutant	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
that	NN	O	O
does	NN	O	O
not	NN	O	O
activate	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
multiple	NN	O	O
stimuli	NN	O	O
.	NN	O	O

Numerous	NN	O	O
genes	NN	O	O
required	NN	O	O
during	NN	O	O
the	NN	O	O
immune	NN	O	O
or	NN	O	O
inflammation	NN	O	O
response	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
adhesion	NN	O	O
process	NN	O	O
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Associated	NN	O	O
with	NN	O	O
its	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
,	NN	O	O
NF-kappaB	NN	O	B-protein
resides	NN	O	O
as	NN	O	O
an	NN	O	O
inactive	NN	O	O
form	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

Upon	NN	O	O
stimulation	NN	O	O
by	NN	O	O
various	NN	O	O
agents	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
is	NN	O	O
proteolyzed	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
translocates	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
,	NN	O	O
where	NN	O	O
it	NN	O	O
activates	NN	O	O
its	NN	O	O
target	NN	O	O
genes	NN	O	O
.	NN	O	O

The	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
inactivation	NN	O	O
remain	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
characterized	NN	O	O
a	NN	O	O
cellular	NN	O	O
mutant	NN	O	O
,	NN	O	O
the	NN	O	O
70/Z3-derived	NN	O	B-cell_line
1.3E2	NN	O	I-cell_line
murine	NN	O	I-cell_line
pre-B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
that	NN	O	O
does	NN	O	O
not	NN	O	O
activate	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
several	NN	O	O
stimuli	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
by	NN	O	O
lipopolysaccharide	NN	O	O
,	NN	O	O
Taxol	NN	O	O
,	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
,	NN	O	O
interleukin-1	NN	O	B-protein
,	NN	O	O
or	NN	O	O
double-stranded	NN	O	B-RNA
RNA	NN	O	I-RNA
,	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
alpha	NN	O	I-protein
is	NN	O	O
not	NN	O	O
degraded	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
induced	NN	O	O
phosphorylation	NN	O	O
on	NN	O	O
serines	NN	O	O
32	NN	O	O
and	NN	O	O
36	NN	O	O
.	NN	O	O

Neither	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
alpha	NN	O	I-protein
nor	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
p50	NN	O	B-protein
or	NN	O	O
relA	NN	O	B-protein
,	NN	O	O
the	NN	O	O
two	NN	O	O
major	NN	O	O
subunits	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
this	NN	O	O
cell	NN	O	O
line	NN	O	O
,	NN	O	O
accounts	NN	O	O
for	NN	O	O
this	NN	O	O
phosphorylation	NN	O	O
defect	NN	O	O
.	NN	O	O

As	NN	O	O
well	NN	O	O
as	NN	O	O
culminating	NN	O	O
in	NN	O	O
the	NN	O	O
inducible	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
alpha	NN	O	O
on	NN	O	O
serines	NN	O	O
32	NN	O	O
and	NN	O	O
36	NN	O	O
,	NN	O	O
all	NN	O	O
the	NN	O	O
stimuli	NN	O	O
that	NN	O	O
are	NN	O	O
inactive	NN	O	O
on	NN	O	O
1.3E2	NN	O	B-cell_line
cells	NN	O	I-cell_line
exhibit	NN	O	O
a	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
the	NN	O	O
antioxidant	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
stimuli	NN	O	O
such	NN	O	O
as	NN	O	O
hyperosmotic	NN	O	O
shock	NN	O	O
or	NN	O	O
phosphatase	NN	O	O
inhibitors	NN	O	O
,	NN	O	O
which	NN	O	O
use	NN	O	O
PDTC-insensitive	NN	O	O
pathways	NN	O	O
,	NN	O	O
induce	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
alpha	NN	O	O
degradation	NN	O	O
in	NN	O	O
1.3E2	NN	O	B-cell_line
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
redox	NN	O	O
status	NN	O	O
of	NN	O	O
1.3E2	NN	O	B-cell_line
does	NN	O	O
not	NN	O	O
reveal	NN	O	O
any	NN	O	O
difference	NN	O	O
from	NN	O	O
wild-type	NN	O	B-cell_line
70Z/3	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
also	NN	O	O
report	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	I-protein
HTLV-1	NN	O	I-protein
)	NN	O	I-protein
-derived	NN	O	I-protein
Tax	NN	O	I-protein
trans-activator	NN	O	I-protein
induces	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
1.3E2	NN	O	B-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
viral	NN	O	O
protein	NN	O	O
does	NN	O	O
not	NN	O	O
operate	NN	O	O
via	NN	O	O
the	NN	O	O
defective	NN	O	O
pathway	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
two	NN	O	O
other	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
beta	NN	O	I-protein
and	NN	O	O
the	NN	O	O
recently	NN	O	O
identified	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
epsilon	NN	O	I-protein
,	NN	O	O
are	NN	O	O
not	NN	O	O
degraded	NN	O	O
in	NN	O	O
the	NN	O	O
1.3E2	NN	O	B-cell_line
cell	NN	O	O
line	NN	O	O
following	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
1.3E2	NN	O	B-cell_line
is	NN	O	O
a	NN	O	O
cellular	NN	O	B-protein
transduction	NN	O	I-protein
mutant	NN	O	I-protein
exhibiting	NN	O	O
a	NN	O	O
defect	NN	O	O
in	NN	O	O
a	NN	O	O
step	NN	O	O
that	NN	O	O
is	NN	O	O
required	NN	O	O
by	NN	O	O
several	NN	O	O
different	NN	O	O
stimuli	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
this	NN	O	O
analysis	NN	O	O
suggests	NN	O	O
a	NN	O	O
common	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
that	NN	O	O
trigger	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
beta	NN	O	I-protein
,	NN	O	O
and	NN	O	O
I	NN	O	B-protein
kappaB	NN	O	I-protein
epsilon	NN	O	I-protein
degradation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
/RelA	NN	O	B-protein
synergy	NN	O	O
in	NN	O	O
distinguishing	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
from	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
-induced	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
transcription	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
required	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
other	NN	O	O
immunological	NN	O	O
functions	NN	O	O
.	NN	O	O

The	NN	O	O
NF-kappa	NN	O	B-DNA
B1	NN	O	I-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
105-kD	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
is	NN	O	O
the	NN	O	O
precursor	NN	O	O
of	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
component	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
we	NN	O	O
and	NN	O	O
others	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
regulates	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B1	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
manuscript	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
stimulated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
phytohemagglutin	NN	O	B-protein
(	NN	O	O
PHA	NN	O	B-protein
)	NN	O	O
display	NN	O	O
significantly	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B1	NN	O	I-protein
encoding	NN	O	O
transcripts	NN	O	B-RNA
than	NN	O	O
cells	NN	O	O
stimulated	NN	O	O
with	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
,	NN	O	O
despite	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
both	NN	O	O
stimuli	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B1	NN	O	I-DNA
promoter	NN	O	I-DNA
identified	NN	O	O
an	NN	O	O
Egr-1	NN	O	B-DNA
site	NN	O	I-DNA
which	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
essential	NN	O	O
for	NN	O	O
both	NN	O	O
the	NN	O	O
PMA/	NN	O	O
PHA	NN	O	B-protein
-mediated	NN	O	O
induction	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
observed	NN	O	O
after	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
RelA	NN	O	O
subunit	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
Egr-1	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
Egr-1	NN	O	B-protein
induction	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
endogenous	NN	O	O
NF-kappa	NN	O	B-DNA
B1	NN	O	I-DNA
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
since	NN	O	O
PMA/	NN	O	O
PHA-stimulated	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
expressing	NN	O	O
antisense	NN	O	O
Egr-1	NN	O	B-RNA
RNA	NN	O	I-RNA
were	NN	O	O
inhibited	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
upregulate	NN	O	O
NF-kappa	NN	O	O
B1	NN	O	O
transcription	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
synergy	NN	O	O
mediated	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
Egr-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
may	NN	O	O
have	NN	O	O
important	NN	O	O
ramifications	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
by	NN	O	O
upregulating	NN	O	O
NF-kappa	NN	O	B-DNA
B1	NN	O	I-DNA
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
synthesis	NN	O	O
and	NN	O	O
immunogenicity	NN	O	O
of	NN	O	O
varicella-zoster	NN	O	B-protein
virus	NN	O	I-protein
glycoprotein	NN	O	I-protein
E	NN	O	I-protein
and	NN	O	O
immediate-early	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
IE62	NN	O	B-protein
)	NN	O	O
expressed	NN	O	O
in	NN	O	O
recombinant	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
virus-1	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
evaluate	NN	O	O
the	NN	O	O
conditions	NN	O	O
for	NN	O	O
optimal	NN	O	O
expression	NN	O	O
and	NN	O	O
immunogenicity	NN	O	O
of	NN	O	O
varicella-zoster	NN	O	B-protein
virus	NN	O	I-protein
(	NN	O	I-protein
VZV	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
a	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
virus-1	NN	O	O
(	NN	O	O
HSV-1	NN	O	O
)	NN	O	O
vector	NN	O	O
,	NN	O	O
we	NN	O	O
selected	NN	O	O
the	NN	O	O
VZV	NN	O	B-protein
glycoprotein	NN	O	I-protein
E	NN	O	I-protein
(	NN	O	O
gE	NN	O	B-protein
)	NN	O	O
,	NN	O	O
encoded	NN	O	O
by	NN	O	O
ORF	NN	O	B-DNA
68	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
VZV	NN	O	B-protein
product	NN	O	I-protein
of	NN	O	O
ORF	NN	O	B-DNA
62	NN	O	I-DNA
,	NN	O	O
an	NN	O	O
immediate-early	NN	O	B-protein
major	NN	O	I-protein
tegument	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
IE62	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Three	NN	O	O
HSV/VZV	NN	O	B-DNA
recombinants	NN	O	I-DNA
were	NN	O	O
generated	NN	O	O
:	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
VZV	NN	O	B-DNA
gE	NN	O	I-DNA
protein	NN	O	I-DNA
coding	NN	O	I-DNA
sequences	NN	O	I-DNA
along	NN	O	O
with	NN	O	O
the	NN	O	O
promoter	NN	O	B-protein
region	NN	O	I-protein
were	NN	O	O
inserted	NN	O	O
into	NN	O	O
the	NN	O	O
thymidine	NN	O	B-DNA
kinase	NN	O	I-DNA
(	NN	O	I-DNA
TK	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
of	NN	O	O
HSV-1	NN	O	O
strain	NN	O	O
KOS	NN	O	O
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
VZV	NN	O	B-protein
gE	NN	O	I-protein
expressed	NN	O	O
from	NN	O	O
the	NN	O	O
HSV-1	NN	O	B-DNA
ICP4	NN	O	I-DNA
promoter	NN	O	I-DNA
was	NN	O	O
inserted	NN	O	O
into	NN	O	O
the	NN	O	O
glycoprotein	NN	O	B-DNA
C	NN	O	I-DNA
(	NN	O	I-DNA
gC	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
of	NN	O	O
HSV-1	NN	O	O
strain	NN	O	O
F	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
VZV	NN	O	B-DNA
IE62	NN	O	I-DNA
protein	NN	O	I-DNA
coding	NN	O	I-DNA
sequences	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
HSV-1	NN	O	B-DNA
ICP4	NN	O	I-DNA
promoter	NN	O	I-DNA
were	NN	O	O
inserted	NN	O	O
into	NN	O	O
the	NN	O	O
gC	NN	O	B-DNA
gene	NN	O	I-DNA
of	NN	O	O
HSV-1	NN	O	O
strain	NN	O	O
F	NN	O	O
.	NN	O	O

Immunoblot	NN	O	O
analysis	NN	O	O
and	NN	O	O
immunoperoxidase	NN	O	O
staining	NN	O	O
of	NN	O	O
infected	NN	O	O
cell	NN	O	O
monolayers	NN	O	O
demonstrated	NN	O	O
vector	NN	O	O
expression	NN	O	O
of	NN	O	O
VZV	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Following	NN	O	O
intracranial	NN	O	O
inoculation	NN	O	O
in	NN	O	O
mice	NN	O	O
,	NN	O	O
both	NN	O	O
VZV	NN	O	B-protein
gE-HSV	NN	O	I-protein
(	NN	O	O
TK	NN	O	B-protein
)	NN	O	O
and	NN	O	O
VZV	NN	O	B-protein
IE62-HSV	NN	O	I-protein
(	NN	O	O
gC	NN	O	B-protein
)	NN	O	O
induced	NN	O	O
an	NN	O	O
IgG	NN	O	B-protein
response	NN	O	O
against	NN	O	O
VZV	NN	O	B-protein
gE	NN	O	I-protein
or	NN	O	O
VZV	NN	O	B-protein
IE62	NN	O	I-protein
.	NN	O	O

When	NN	O	O
tested	NN	O	O
in	NN	O	O
cytotoxicity	NN	O	O
assays	NN	O	O
using	NN	O	O
T-lymphocytes	NN	O	B-cell_type
from	NN	O	O
VZV	NN	O	O
immune	NN	O	O
human	NN	O	O
donors	NN	O	O
,	NN	O	O
the	NN	O	O
range	NN	O	O
of	NN	O	O
precursor	NN	O	O
frequencies	NN	O	O
for	NN	O	O
T-lymphocytes	NN	O	B-cell_type
that	NN	O	O
recognized	NN	O	O
VZV	NN	O	B-protein
gE	NN	O	I-protein
or	NN	O	O
VZV	NN	O	B-protein
IE62	NN	O	I-protein
was	NN	O	O
similar	NN	O	O
whether	NN	O	O
these	NN	O	O
proteins	NN	O	O
were	NN	O	O
expressed	NN	O	O
by	NN	O	O
HSV-1	NN	O	O
or	NN	O	O
a	NN	O	O
vaccinia	NN	O	O
vector	NN	O	O
.	NN	O	O

These	NN	O	O
experiments	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
HSV-1	NN	O	O
is	NN	O	O
a	NN	O	O
competent	NN	O	O
vector	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
VZV	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
support	NN	O	O
the	NN	O	O
feasibility	NN	O	O
of	NN	O	O
engineering	NN	O	O
a	NN	O	O
combined	NN	O	O
vaccine	NN	O	O
for	NN	O	O
these	NN	O	O
closely	NN	O	O
related	NN	O	O
alpha-herpesviruses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
dephosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
,	NN	O	O
NF-AT1	NN	O	B-protein
,	NN	O	O
in	NN	O	O
Tax-expressing	NN	O	B-cell_type
and	NN	O	I-cell_type
type	NN	O	I-cell_type
I	NN	O	I-cell_type
human	NN	O	I-cell_type
T-cell	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
virus-infected	NN	O	I-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
tax	NN	O	O
gene	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
type	NN	O	O
I	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
transactivates	NN	O	O
interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
through	NN	O	O
activation	NN	O	O
of	NN	O	O
an	NN	O	O
enhancer	NN	O	O
termed	NN	O	O
CD28	NN	O	B-DNA
responsive	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CD28RE	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Tax	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
the	NN	O	O
CD28RE	NN	O	B-DNA
is	NN	O	O
partially	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
,	NN	O	O
NF-AT1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
NF-AT1	NN	O	B-protein
is	NN	O	O
constitutively	NN	O	O
active	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
Tax	NN	O	B-DNA
cDNA	NN	O	I-DNA
,	NN	O	O
although	NN	O	O
the	NN	O	O
underlying	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
and	NN	O	O
physiological	NN	O	O
relevance	NN	O	O
of	NN	O	O
this	NN	O	O
finding	NN	O	O
remain	NN	O	O
unclear	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
active	NN	O	O
form	NN	O	O
of	NN	O	O
NF-AT1	NN	O	B-protein
is	NN	O	O
also	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
HTLV-I-transformed	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
express	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-AT1	NN	O	B-protein
in	NN	O	O
these	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
its	NN	O	O
dephosphorylation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
dephosphorylated	NN	O	O
NF-AT1	NN	O	B-protein
can	NN	O	O
be	NN	O	O
rapidly	NN	O	O
rephosphorylated	NN	O	O
when	NN	O	O
the	NN	O	O
cells	NN	O	O
are	NN	O	O
incubated	NN	O	O
with	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
an	NN	O	O
immunosuppressant	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
serine/threonine	NN	O	B-protein
phosphatase	NN	O	I-protein
calcineurin	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-AT1	NN	O	B-protein
in	NN	O	O
Tax-expressing	NN	O	B-cell_line
and	NN	O	I-cell_line
HTLV-I-transformed	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
results	NN	O	O
from	NN	O	O
its	NN	O	O
dephosphorylation	NN	O	O
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
deregulation	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
NFAT-family	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NFAT	NN	O	B-protein
proteins	NN	O	I-protein
constitute	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
mediating	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
specific	NN	O	O
antisera	NN	O	O
in	NN	O	O
immunoprecipitation	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
NFATp	NN	O	B-protein
(	NN	O	O
135	NN	O	B-protein
kDa	NN	O	I-protein
)	NN	O	O
is	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
synthesis	NN	O	O
of	NN	O	O
NFATc	NN	O	B-protein
(	NN	O	O
predominant	NN	O	O
form	NN	O	O
of	NN	O	O
86	NN	O	B-protein
kDa	NN	O	I-protein
)	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
ionomycin	NN	O	O
treatment	NN	O	O
.	NN	O	O

NFAT4/x	NN	O	B-protein
was	NN	O	O
very	NN	O	O
weakly	NN	O	O
expressed	NN	O	O
in	NN	O	O
unstimulated	NN	O	O
cells	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
level	NN	O	O
did	NN	O	O
not	NN	O	O
increase	NN	O	O
upon	NN	O	O
treatment	NN	O	O
with	NN	O	O
activating	NN	O	O
agents	NN	O	O
.	NN	O	O

NFAT3	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
not	NN	O	O
observed	NN	O	O
under	NN	O	O
any	NN	O	O
conditions	NN	O	O
.	NN	O	O

Higher-molecular-weight	NN	O	B-protein
species	NN	O	I-protein
of	NN	O	O
NFATc	NN	O	B-protein
(	NN	O	O
of	NN	O	O
110	NN	O	O
and	NN	O	O
140	NN	O	O
kDa	NN	O	O
)	NN	O	O
were	NN	O	O
also	NN	O	O
detected	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
translation	NN	O	O
of	NN	O	O
NFATc	NN	O	B-RNA
mRNA	NN	O	I-RNA
apparently	NN	O	O
initiates	NN	O	O
at	NN	O	O
two	NN	O	O
different	NN	O	O
AUG	NN	O	B-DNA
codons	NN	O	I-DNA
,	NN	O	O
giving	NN	O	O
rise	NN	O	O
to	NN	O	O
proteins	NN	O	O
that	NN	O	O
differ	NN	O	O
in	NN	O	O
size	NN	O	O
by	NN	O	O
36	NN	O	O
amino	NN	O	O
acids	NN	O	O
.	NN	O	O

Additional	NN	O	O
size	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
both	NN	O	O
NFATc	NN	O	B-protein
and	NN	O	O
NFATp	NN	O	B-protein
results	NN	O	O
from	NN	O	O
phosphorylation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
ionomycin	NN	O	O
treatment	NN	O	O
,	NN	O	O
exposure	NN	O	O
of	NN	O	O
cells	NN	O	O
to	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
plus	NN	O	O
anti-CD28	NN	O	B-protein
did	NN	O	O
not	NN	O	O
induce	NN	O	O
NFATc	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
under	NN	O	O
these	NN	O	O
conditions	NN	O	O
,	NN	O	O
interleukin-2	NN	O	B-protein
synthesis	NN	O	O
by	NN	O	O
these	NN	O	O
cells	NN	O	O
is	NN	O	O
apparently	NN	O	O
independent	NN	O	O
of	NN	O	O
NFATc	NN	O	B-protein
.	NN	O	O

In	NN	O	O
DNA	NN	O	O
binding	NN	O	O
assays	NN	O	O
,	NN	O	O
both	NN	O	O
PMA	NN	O	O
plus	NN	O	O
anti-CD28	NN	O	B-protein
and	NN	O	O
PMA	NN	O	O
plus	NN	O	O
ionomycin	NN	O	O
resulted	NN	O	O
in	NN	O	O
nuclear	NN	O	B-protein
NFAT	NN	O	I-protein
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
the	NN	O	O
PMA-ionomycin-induced	NN	O	O
synthesis	NN	O	O
of	NN	O	O
NFATc	NN	O	B-protein
that	NN	O	O
was	NN	O	O
detected	NN	O	O
by	NN	O	O
immunoprecipitation	NN	O	O
was	NN	O	O
not	NN	O	O
mirrored	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
assays	NN	O	O
:	NN	O	O
nearly	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
activity	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
NFATp	NN	O	B-protein
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
first	NN	O	O
study	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
all	NN	O	O
family	NN	O	O
members	NN	O	O
at	NN	O	O
the	NN	O	O
protein	NN	O	O
level	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Of	NN	O	O
the	NN	O	O
GATA-binding	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
only	NN	O	O
GATA-4	NN	O	B-protein
selectively	NN	O	O
regulates	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-5	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
IL-5	NN	O	B-cell_type
producing	NN	O	I-cell_type
cells	NN	O	I-cell_type
which	NN	O	O
express	NN	O	O
multiple	NN	O	B-protein
GATA-binding	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Interleukin-5	NN	O	B-protein
(	NN	O	O
IL-5	NN	O	B-protein
)	NN	O	O
is	NN	O	O
produced	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
known	NN	O	O
to	NN	O	O
support	NN	O	O
B	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
eosinophilic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
progenitor	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
ATL-16T	NN	O	B-cell_line
cells	NN	O	I-cell_line
which	NN	O	O
express	NN	O	O
IL-5	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
region	NN	O	O
,	NN	O	O
within	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-5	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
that	NN	O	O
regulates	NN	O	O
IL-5	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

This	NN	O	O
cis-acting	NN	O	B-DNA
sequence	NN	O	I-DNA
contains	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
,	NN	O	O
(	NN	O	O
A/T	NN	O	O
)	NN	O	O
GATA	NN	O	O
(	NN	O	O
A/G	NN	O	O
)	NN	O	O
,	NN	O	O
for	NN	O	O
GATA-binding	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
thus	NN	O	O
suggests	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
these	NN	O	O
family	NN	O	O
members	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
cloning	NN	O	O
of	NN	O	O
human	NN	O	B-protein
GATA-4	NN	O	I-protein
(	NN	O	O
hGATA-4	NN	O	B-protein
)	NN	O	O
and	NN	O	O
show	NN	O	O
that	NN	O	O
hGATA-4	NN	O	B-protein
selectively	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
-70	NN	O	B-DNA
GATA	NN	O	I-DNA
site	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
proximal	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

By	NN	O	O
promoter	NN	O	O
deletion	NN	O	O
and	NN	O	O
mutation	NN	O	O
analyses	NN	O	O
,	NN	O	O
we	NN	O	O
established	NN	O	O
this	NN	O	O
region	NN	O	O
as	NN	O	O
a	NN	O	O
positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Cotransfection	NN	O	O
experiments	NN	O	O
revealed	NN	O	O
that	NN	O	O
both	NN	O	O
hGATA-4	NN	O	B-protein
and	NN	O	O
PMA/A23187	NN	O	O
stimulation	NN	O	O
are	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

The	NN	O	O
requirement	NN	O	O
of	NN	O	O
another	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
called	NN	O	O
CLE0	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
lies	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
-70	NN	O	B-DNA
GATA	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
also	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

ATL-16T	NN	O	B-cell_line
cells	NN	O	I-cell_line
express	NN	O	O
mRNA	NN	O	B-RNA
of	NN	O	O
three	NN	O	O
GATA-binding	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
hGATA-2	NN	O	B-protein
,	NN	O	O
hGATA-3	NN	O	B-protein
and	NN	O	O
hGATA-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
each	NN	O	O
of	NN	O	O
them	NN	O	O
has	NN	O	O
a	NN	O	O
potential	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
(	NN	O	I-DNA
A/T	NN	O	I-DNA
)	NN	O	I-DNA
GATA	NN	O	I-DNA
(	NN	O	I-DNA
G/	NN	O	I-DNA
A	NN	O	I-DNA
)	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
using	NN	O	O
ATL-16T	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
GATA-4	NN	O	B-protein
is	NN	O	O
the	NN	O	O
only	NN	O	O
GATA-binding	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
forms	NN	O	O
specific	NN	O	O
DNA-protein	NN	O	B-protein
complex	NN	O	I-protein
with	NN	O	O
the	NN	O	O
-70	NN	O	B-DNA
GATA	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
with	NN	O	O
extracts	NN	O	O
of	NN	O	O
COS	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
GATA-binding	NN	O	B-protein
proteins	NN	O	I-protein
showed	NN	O	O
that	NN	O	O
GATA-4	NN	O	B-protein
has	NN	O	O
the	NN	O	O
highest	NN	O	O
binding	NN	O	O
affinity	NN	O	O
to	NN	O	O
the	NN	O	O
-70	NN	O	B-DNA
GATA	NN	O	I-DNA
site	NN	O	I-DNA
among	NN	O	O
the	NN	O	O
three	NN	O	O
GATA-binding	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

When	NN	O	O
the	NN	O	O
transactivation	NN	O	O
ability	NN	O	O
was	NN	O	O
compared	NN	O	O
among	NN	O	O
the	NN	O	O
three	NN	O	O
,	NN	O	O
GATA-4	NN	O	B-protein
showed	NN	O	O
the	NN	O	O
highest	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
selective	NN	O	O
role	NN	O	O
of	NN	O	O
GATA-4	NN	O	B-protein
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
circumstance	NN	O	O
where	NN	O	O
multiple	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
GATA-binding	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
expressed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Control	NN	O	O
of	NN	O	O
NFATx1	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
by	NN	O	O
a	NN	O	O
calcineurin-regulated	NN	O	B-protein
inhibitory	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

The	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
regulates	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
located	NN	O	O
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
several	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

NFATx1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
preferentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
four	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
performed	NN	O	O
domain	NN	O	O
analysis	NN	O	O
of	NN	O	O
NFATx1	NN	O	B-protein
by	NN	O	O
examining	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
deletion	NN	O	O
mutations	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
NFATx1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
interaction	NN	O	O
with	NN	O	O
AP-1	NN	O	O
polypeptides	NN	O	O
were	NN	O	O
dependent	NN	O	O
on	NN	O	O
its	NN	O	O
central	NN	O	B-protein
Rel	NN	O	I-protein
similarity	NN	O	I-protein
region	NN	O	I-protein
and	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
was	NN	O	O
reduced	NN	O	O
by	NN	O	O
deletions	NN	O	O
of	NN	O	O
either	NN	O	O
its	NN	O	O
N-terminal	NN	O	B-protein
domain	NN	O	I-protein
or	NN	O	O
its	NN	O	O
C-terminal	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
intrinsic	NN	O	B-protein
transcriptional	NN	O	I-protein
activation	NN	O	I-protein
motifs	NN	O	I-protein
in	NN	O	O
both	NN	O	O
regions	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
identified	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitory	NN	O	B-protein
sequence	NN	O	I-protein
within	NN	O	O
its	NN	O	O
N-terminal	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
inhibition	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
calcium-calmodulin-dependent	NN	O	B-protein
phosphatase	NN	O	I-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
calcineurin	NN	O	B-protein
associated	NN	O	O
with	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
NFATx1	NN	O	B-protein
at	NN	O	O
multiple	NN	O	B-protein
docking	NN	O	I-protein
sites	NN	O	I-protein
and	NN	O	O
caused	NN	O	O
a	NN	O	O
reduction	NN	O	O
of	NN	O	O
size	NN	O	O
,	NN	O	O
indicative	NN	O	O
of	NN	O	O
dephosphorylation	NN	O	O
,	NN	O	O
in	NN	O	O
NFATx1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
mapped	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
activity	NN	O	O
to	NN	O	O
less	NN	O	O
than	NN	O	O
60	NN	O	B-protein
residues	NN	O	I-protein
,	NN	O	O
containing	NN	O	O
motifs	NN	O	O
that	NN	O	O
are	NN	O	O
conserved	NN	O	O
in	NN	O	O
all	NN	O	O
NFAT	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
deletion	NN	O	O
in	NN	O	O
NFATx1	NN	O	B-protein
of	NN	O	O
the	NN	O	O
mapped	NN	O	O
60	NN	O	B-protein
residues	NN	O	I-protein
leads	NN	O	O
to	NN	O	O
its	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
independent	NN	O	O
of	NN	O	O
calcium	NN	O	O
signaling	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
model	NN	O	O
proposing	NN	O	O
that	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
domain	NN	O	I-protein
confers	NN	O	O
calcium-signaling	NN	O	O
dependence	NN	O	O
on	NN	O	O
NFATx1	NN	O	B-protein
transactivation	NN	O	O
activity	NN	O	O
by	NN	O	O
regulating	NN	O	O
its	NN	O	O
intracellular	NN	O	O
localization	NN	O	O
through	NN	O	O
a	NN	O	O
protein	NN	O	O
module	NN	O	O
that	NN	O	O
associates	NN	O	O
with	NN	O	O
calcineurin	NN	O	B-protein
and	NN	O	O
is	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
its	NN	O	O
phosphatase	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Neuronal	NN	O	B-protein
(	NN	O	I-protein
type	NN	O	I-protein
I	NN	O	I-protein
)	NN	O	I-protein
nitric	NN	O	I-protein
oxide	NN	O	I-protein
synthase	NN	O	I-protein
regulates	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
activity	NN	O	O
and	NN	O	O
immunologic	NN	O	B-protein
(	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
)	NN	O	I-protein
nitric	NN	O	I-protein
oxide	NN	O	I-protein
synthase	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Nitric	NN	O	O
oxide	NN	O	O
subserves	NN	O	O
diverse	NN	O	O
physiologic	NN	O	O
roles	NN	O	O
in	NN	O	O
the	NN	O	O
nervous	NN	O	O
system	NN	O	O
.	NN	O	O

NO	NN	O	O
is	NN	O	O
produced	NN	O	O
from	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
different	NN	O	O
NO	NN	O	B-protein
synthase	NN	O	I-protein
(	NN	O	I-protein
NOS	NN	O	I-protein
)	NN	O	I-protein
isoforms	NN	O	I-protein
:	NN	O	O
neuronal	NN	O	B-protein
NOS	NN	O	I-protein
(	NN	O	O
nNOS	NN	O	B-protein
)	NN	O	O
,	NN	O	O
endothelial	NN	O	B-protein
NOS	NN	O	I-protein
,	NN	O	O
and	NN	O	O
immunologic	NN	O	B-protein
NOS	NN	O	I-protein
(	NN	O	O
iNOS	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
nNOS	NN	O	B-protein
is	NN	O	O
the	NN	O	O
predominant	NN	O	O
isoform	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
glia	NN	O	B-cell_type
.	NN	O	O

NO	NN	O	O
derived	NN	O	O
from	NN	O	O
nNOS	NN	O	B-protein
in	NN	O	O
glia	NN	O	B-cell_type
inhibits	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF	NN	O	B-protein
kappaB	NN	O	I-protein
)	NN	O	O
as	NN	O	O
NOS	NN	O	O
inhibitors	NN	O	O
enhance	NN	O	O
basal	NN	O	O
NF	NN	O	B-protein
kappaB	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappaB	NN	O	I-protein
in	NN	O	O
most	NN	O	O
cells	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
PDTC	NN	O	O
is	NN	O	O
also	NN	O	O
a	NN	O	O
potent	NN	O	O
scavenger	NN	O	O
of	NN	O	O
NO	NN	O	O
through	NN	O	O
formation	NN	O	O
of	NN	O	O
mononitrosyl	NN	O	O
iron	NN	O	O
complexes	NN	O	O
with	NN	O	O
PDTC	NN	O	O
.	NN	O	O

In	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
human	NN	O	O
T-cell	NN	O	B-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
induces	NN	O	O
NF	NN	O	B-protein
kappaB	NN	O	I-protein
activation	NN	O	O
that	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
PDTC	NN	O	O
.	NN	O	O

Contrary	NN	O	O
to	NN	O	O
the	NN	O	O
results	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
PDTC	NN	O	O
did	NN	O	O
not	NN	O	O
inhibit	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
-induced	NN	O	O
NF	NN	O	B-protein
kappaB	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
astrocytes	NN	O	B-cell_type
;	NN	O	O
instead	NN	O	O
PDTC	NN	O	O
itself	NN	O	O
induces	NN	O	O
NF	NN	O	B-protein
kappaB	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
astrocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
this	NN	O	O
may	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
scavenging	NN	O	O
of	NN	O	O
endogenously	NN	O	O
produced	NN	O	O
NO	NN	O	O
by	NN	O	O
the	NN	O	O
PDTC	NN	O	O
iron	NN	O	O
complex	NN	O	O
.	NN	O	O

In	NN	O	O
astrocytes	NN	O	B-cell_type
PDTC	NN	O	O
also	NN	O	O
dramatically	NN	O	O
induces	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappaB-dependent	NN	O	I-protein
enzyme	NN	O	I-protein
,	NN	O	O
iNOS	NN	O	B-protein
,	NN	O	O
supporting	NN	O	O
the	NN	O	O
physiologic	NN	O	O
relevance	NN	O	O
of	NN	O	O
endogenous	NN	O	O
NO	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappaB	NN	O	I-protein
.	NN	O	O

NF	NN	O	B-protein
kappaB	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
glia	NN	O	B-cell_type
from	NN	O	O
mice	NN	O	O
lacking	NN	O	O
nNOS	NN	O	B-protein
responds	NN	O	O
more	NN	O	O
rapidly	NN	O	O
to	NN	O	O
PDTC	NN	O	O
compared	NN	O	O
with	NN	O	O
astrocytes	NN	O	B-cell_type
from	NN	O	O
wild-type	NN	O	O
mice	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
nNOS	NN	O	B-protein
in	NN	O	O
astrocytes	NN	O	B-cell_type
regulates	NN	O	O
NF	NN	O	B-protein
kappaB	NN	O	I-protein
activity	NN	O	O
and	NN	O	O
iNOS	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
indicate	NN	O	O
a	NN	O	O
novel	NN	O	O
regulatory	NN	O	O
role	NN	O	O
for	NN	O	O
nNOS	NN	O	B-protein
in	NN	O	O
tonically	NN	O	O
suppressing	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
,	NN	O	O
NF	NN	O	B-DNA
kappaB-regulated	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
in	NN	O	O
interleukin-6	NN	O	B-protein
-induced	NN	O	O
IgM	NN	O	B-protein
production	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Interleukin-6	NN	O	B-protein
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
B-cell	NN	O	B-protein
growth	NN	O	I-protein
and	NN	O	I-protein
differentiation	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

IL-6	NN	O	B-protein
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
SKW6.4	NN	O	B-cell_line
,	NN	O	O
leads	NN	O	O
to	NN	O	O
increased	NN	O	O
IgM	NN	O	B-protein
production	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
IL-6	NN	O	B-protein
induces	NN	O	O
activation	NN	O	O
of	NN	O	O
JAK1	NN	O	B-protein
and	NN	O	O
JAK2	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
chimeric	NN	O	O
IL-6	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
comprised	NN	O	O
of	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
tail	NN	O	I-protein
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-protein
receptor	NN	O	I-protein
subunit	NN	O	I-protein
gp130	NN	O	B-protein
fused	NN	O	O
to	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
epidermal	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
EGF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
was	NN	O	O
stably	NN	O	O
transfected	NN	O	O
into	NN	O	O
SKW6.4	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

EGF	NN	O	O
treatment	NN	O	O
induced	NN	O	O
IgM	NN	O	B-protein
production	NN	O	O
in	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
an	NN	O	O
intact	NN	O	O
gp130	NN	O	B-protein
cytoplasmic	NN	O	I-protein
tail	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
untransfected	NN	O	B-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	B-protein
tail	NN	O	I-protein
lacking	NN	O	O
all	NN	O	O
four	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
Stat	NN	O	I-protein
)	NN	O	I-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
EGF	NN	O	O
treatment	NN	O	O
induced	NN	O	O
Stat3	NN	O	B-protein
phosphorylation	NN	O	O
in	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
intact	NN	O	O
chimeric	NN	O	B-protein
EGF-gp130	NN	O	I-protein
receptor	NN	O	I-protein
along	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
DNA-mobility	NN	O	O
shift	NN	O	O
of	NN	O	O
a	NN	O	O
classical	NN	O	B-DNA
interferon-gamma-activated	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
define	NN	O	O
further	NN	O	O
the	NN	O	O
relation	NN	O	O
between	NN	O	O
Stat3	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
enhanced	NN	O	O
IgM	NN	O	B-protein
production	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
chimeric	NN	O	O
gp130	NN	O	O
on	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
genetic	NN	O	O
element	NN	O	O
linked	NN	O	O
to	NN	O	O
immunoglobulin	NN	O	B-protein
production	NN	O	O
,	NN	O	O
namely	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
(	NN	O	O
IgH-enhancer	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Parental	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
transfected	NN	O	O
SKW6.4	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
with	NN	O	O
an	NN	O	O
IgH-enhancer-luciferase	NN	O	B-DNA
construct	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
IgH-luciferase	NN	O	B-DNA
construct	NN	O	I-DNA
was	NN	O	O
induced	NN	O	O
upon	NN	O	O
ligation	NN	O	O
of	NN	O	O
the	NN	O	O
full-length	NN	O	B-protein
chimeric	NN	O	I-protein
receptor	NN	O	I-protein
but	NN	O	O
not	NN	O	O
by	NN	O	O
truncated	NN	O	B-protein
gp130	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
gp130	NN	O	B-protein
-induced	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
abrogated	NN	O	O
by	NN	O	O
Stat3EE	NN	O	B-protein
,	NN	O	O
a	NN	O	O
mutant	NN	O	O
Stat3	NN	O	B-protein
incapable	NN	O	O
of	NN	O	O
binding	NN	O	O
DNA	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-6	NN	O	B-protein
-induced	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
IgM	NN	O	B-protein
production	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
controlled	NN	O	O
by	NN	O	O
Stat3	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

T-lymphocytes	NN	O	B-cell_type
from	NN	O	O
individuals	NN	O	O
with	NN	O	O
filarial	NN	O	O
inflammatory	NN	O	O
disease	NN	O	O
have	NN	O	O
increased	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
in	NN	O	O
vitro	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
of	NN	O	O
circulating	NN	O	B-cell_type
filarial	NN	O	I-cell_type
antigen-specific	NN	O	I-cell_type
T-cells	NN	O	I-cell_type
was	NN	O	O
examined	NN	O	O
in	NN	O	O
Wuchereria	NN	O	O
banerofti	NN	O	O
infection	NN	O	O
.	NN	O	O

Circulating	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
from	NN	O	O
individuals	NN	O	O
with	NN	O	O
filaria-induced	NN	O	O
lymphatic	NN	O	O
pathology	NN	O	O
(	NN	O	O
LP	NN	O	O
)	NN	O	O
had	NN	O	O
significantly	NN	O	O
greater	NN	O	O
migration	NN	O	O
through	NN	O	O
unstimulated	NN	O	O
HUVEC	NN	O	B-cell_type
monolayers	NN	O	I-cell_type
than	NN	O	O
did	NN	O	O
T-cells	NN	O	O
from	NN	O	O
asymptomatic	NN	O	O
infected	NN	O	O
(	NN	O	O
MF	NN	O	O
)	NN	O	O
individuals	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.04	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
MF	NN	O	O
individuals	NN	O	O
where	NN	O	O
no	NN	O	O
effect	NN	O	O
was	NN	O	O
seen	NN	O	O
,	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
of	NN	O	O
48-hr	NN	O	O
filarial	NN	O	B-cell_type
antigen	NN	O	I-cell_type
stimulated	NN	O	I-cell_type
T-cells	NN	O	I-cell_type
from	NN	O	O
LP	NN	O	O
individuals	NN	O	O
was	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.01	NN	O	O
)	NN	O	O
greater	NN	O	O
than	NN	O	O
migration	NN	O	O
of	NN	O	O
48-hr	NN	O	O
media-stimulated	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
six	NN	O	O
of	NN	O	O
seven	NN	O	O
patients	NN	O	O
examined	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
VLA-4/	NN	O	O
VCAM-1	NN	O	B-protein
pathway	NN	O	O
resulted	NN	O	O
in	NN	O	O
greater	NN	O	O
than	NN	O	O
50	NN	O	O
%	NN	O	O
inhibition	NN	O	O
of	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
of	NN	O	O
T-cells	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
ferritin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
gene	NN	O	I-DNA
:	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
CCAAT	NN	O	B-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
NF-Y	NN	O	I-protein
is	NN	O	O
modulated	NN	O	O
in	NN	O	O
heme-treated	NN	O	B-cell_line
Friend	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
during	NN	O	O
monocyte-to-macrophage	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
ferritin	NN	O	B-DNA
H-chain	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
regulation	NN	O	O
was	NN	O	O
analyzed	NN	O	O
in	NN	O	O
heme-treated	NN	O	B-cell_line
Friend	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
FLCs	NN	O	B-cell_line
)	NN	O	O
and	NN	O	O
during	NN	O	O
monocyte-to-macrophage	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
studied	NN	O	O
,	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
ferritin	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
exerted	NN	O	O
mostly	NN	O	O
at	NN	O	O
the	NN	O	O
translational	NN	O	O
level	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
differentiating	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
must	NN	O	O
incorporate	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
iron	NN	O	O
to	NN	O	O
sustain	NN	O	O
hemoglobin	NN	O	B-protein
synthesis	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
which	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
iron	NN	O	O
storage	NN	O	O
,	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
seemed	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
relevant	NN	O	O
mechanism	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
minimum	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
ferritin	NN	O	B-DNA
H-gene	NN	O	I-DNA
promoter	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
confer	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
by	NN	O	O
heme	NN	O	O
in	NN	O	O
FLCs	NN	O	B-cell_line
to	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
77	NN	O	B-DNA
nucleotides	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
cis	NN	O	B-DNA
element	NN	O	I-DNA
binds	NN	O	O
a	NN	O	O
protein	NN	O	O
complex	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
HRF	NN	O	B-protein
(	NN	O	O
heme-responsive	NN	O	B-protein
factor	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
greatly	NN	O	O
enhanced	NN	O	O
both	NN	O	O
in	NN	O	O
heme-treated	NN	O	O
FLCs	NN	O	B-cell_line
and	NN	O	O
during	NN	O	O
monocyte-to-macrophage	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
CCAAT	NN	O	B-DNA
element	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
reverse	NN	O	O
orientation	NN	O	O
in	NN	O	O
this	NN	O	O
promoter	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
ferritin	NN	O	B-DNA
H-chain	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
binding	NN	O	O
and	NN	O	O
for	NN	O	O
gene	NN	O	O
activity	NN	O	O
,	NN	O	O
since	NN	O	O
a	NN	O	O
single	NN	O	O
point	NN	O	O
mutation	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
abolish	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
HRF	NN	O	B-protein
and	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
transfected	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

By	NN	O	O
competition	NN	O	O
experiments	NN	O	O
and	NN	O	O
supershift	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
the	NN	O	O
induced	NN	O	O
HRF	NN	O	B-protein
as	NN	O	O
containing	NN	O	O
at	NN	O	O
least	NN	O	O
the	NN	O	O
ubiquitous	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-Y	NN	O	B-protein
.	NN	O	O

NF-Y	NN	O	B-protein
is	NN	O	O
formed	NN	O	O
by	NN	O	O
three	NN	O	O
subunits	NN	O	B-protein
,	NN	O	O
A	NN	O	B-protein
,	NN	O	O
B	NN	O	B-protein
,	NN	O	O
and	NN	O	O
C	NN	O	B-protein
,	NN	O	O
all	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
necessary	NN	O	O
for	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

Cotransfection	NN	O	O
with	NN	O	O
a	NN	O	O
transdominant	NN	O	B-protein
negative	NN	O	I-protein
mutant	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NF-YA	NN	O	B-protein
subunit	NN	O	I-protein
abolishes	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
heme	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
NF-Y	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
in	NN	O	O
this	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
also	NN	O	O
observed	NN	O	O
a	NN	O	O
differential	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
NF-YA	NN	O	B-protein
subunit	NN	O	I-protein
in	NN	O	O
heme-treated	NN	O	O
and	NN	O	O
control	NN	O	O
FLCs	NN	O	B-cell_line
and	NN	O	O
during	NN	O	O
monocyte-to-macrophage	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Concomitant	NN	O	O
downregulation	NN	O	O
of	NN	O	O
IgH	NN	O	B-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
and	NN	O	O
c-myc	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
a	NN	O	O
plasmacytoma	NN	O	O
x	NN	O	O
fibroblast	NN	O	O
environment	NN	O	O
:	NN	O	O
implications	NN	O	O
for	NN	O	O
dysregulation	NN	O	O
of	NN	O	O
translocated	NN	O	O
c-myc	NN	O	B-DNA
.	NN	O	O

Regulation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
(	NN	O	I-DNA
IgH	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
B	NN	O	B-DNA
cell-specific	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
intronic	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
additional	NN	O	O
B	NN	O	B-DNA
cell-specific	NN	O	I-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
identified	NN	O	O
recently	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	O
'	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
locus	NN	O	I-DNA
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
latter	NN	O	O
elements	NN	O	O
,	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
of	NN	O	O
particular	NN	O	O
interest	NN	O	O
:	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
it	NN	O	O
is	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
and	NN	O	O
active	NN	O	O
only	NN	O	O
in	NN	O	O
late	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
in	NN	O	O
rodent	NN	O	O
plasmacytomas	NN	O	O
and	NN	O	O
in	NN	O	O
some	NN	O	O
human	NN	O	O
Burkitt	NN	O	O
's	NN	O	O
lymphomas	NN	O	O
it	NN	O	O
is	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
locus	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
LCR	NN	O	B-DNA
)	NN	O	O
that	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
deregulation	NN	O	O
of	NN	O	O
the	NN	O	O
c-myc	NN	O	B-DNA
oncogene	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
translocation	NN	O	O
into	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
locus	NN	O	I-DNA
;	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
Ig	NN	O	B-DNA
gene	NN	O	I-DNA
class	NN	O	I-DNA
switch	NN	O	O
recombination	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
a	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybridization	NN	O	O
approach	NN	O	O
to	NN	O	O
genetically	NN	O	O
analyse	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
mouse	NN	O	B-cell_line
MPC11	NN	O	I-cell_line
plasmacytoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
active	NN	O	O
,	NN	O	O
are	NN	O	O
fused	NN	O	O
with	NN	O	O
fibroblasts	NN	O	B-cell_type
,	NN	O	O
Ig	NN	O	O
expression	NN	O	O
is	NN	O	O
extinguished	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
a	NN	O	O
MPC11	NN	O	B-cell_line
plasmacytoma	NN	O	I-cell_line
x	NN	O	I-cell_line
fibroblast	NN	O	I-cell_line
environment	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
transcriptionally	NN	O	O
inactive	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
several	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
essential	NN	O	O
for	NN	O	O
IgH	NN	O	B-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
,	NN	O	O
is	NN	O	O
lacking	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
explain	NN	O	O
3	NN	O	O
'	NN	O	O
enhancer	NN	O	O
inactivity	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
repressors	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
excluded	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
high	NN	O	O
expression	NN	O	O
level	NN	O	O
of	NN	O	O
c-myc	NN	O	B-DNA
,	NN	O	O
characteristic	NN	O	O
of	NN	O	O
the	NN	O	O
parental	NN	O	O
MPC11	NN	O	B-cell_line
cells	NN	O	I-cell_line
carrying	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
12	NN	O	I-DNA
;	NN	O	I-DNA
15	NN	O	I-DNA
)	NN	O	I-DNA
translocation	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
down-regulated	NN	O	O
in	NN	O	O
the	NN	O	O
hybrids	NN	O	O
to	NN	O	O
that	NN	O	O
in	NN	O	O
unfused	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
multifactorial	NN	O	O
process	NN	O	O
affecting	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
control	NN	O	O
the	NN	O	O
cell-specific	NN	O	O
and	NN	O	O
developmental	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
a	NN	O	O
T-lymphocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
by	NN	O	O
hypochlorous	NN	O	O
acid	NN	O	O
.	NN	O	O

Reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
such	NN	O	O
as	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
serve	NN	O	O
as	NN	O	O
second	NN	O	O
messengers	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
hence	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
and	NN	O	O
replication	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

During	NN	O	O
inflammatory	NN	O	O
reactions	NN	O	O
,	NN	O	O
many	NN	O	O
oxidative	NN	O	O
species	NN	O	O
are	NN	O	O
produced	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
is	NN	O	O
hypochlorous	NN	O	O
acid	NN	O	O
(	NN	O	O
HOCl	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
microbicidal	NN	O	O
effects	NN	O	O
of	NN	O	O
activated	NN	O	O
human	NN	O	B-cell_type
polymorphonuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
a	NN	O	O
T-lymphocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
with	NN	O	O
micromolar	NN	O	O
concentrations	NN	O	O
of	NN	O	O
HOCl	NN	O	O
promoted	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
(	NN	O	O
the	NN	O	O
heterodimer	NN	O	B-protein
p50/p65	NN	O	I-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
,	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
inhibitory	NN	O	I-protein
subunits	NN	O	I-protein
(	NN	O	O
IkappaB	NN	O	B-protein
)	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
both	NN	O	O
IkappaB	NN	O	B-protein
-alpha	NN	O	O
proteolysis	NN	O	O
and	NN	O	O
p105	NN	O	B-protein
processing	NN	O	O
were	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
treatment	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
very	NN	O	O
effective	NN	O	O
when	NN	O	O
cells	NN	O	O
were	NN	O	O
subjected	NN	O	O
to	NN	O	O
hyperthermia	NN	O	O
before	NN	O	O
being	NN	O	O
treated	NN	O	O
with	NN	O	O
HOCl	NN	O	O
.	NN	O	O

Various	NN	O	O
antioxidants	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
,	NN	O	O
p-bromophenacyl-bromide	NN	O	O
and	NN	O	O
nordihydroguaiaretic	NN	O	O
acid	NN	O	O
could	NN	O	O
strongly	NN	O	O
reduce	NN	O	O
NF-kappaB	NN	O	B-protein
translocation	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
oxidative	NN	O	O
species	NN	O	O
in	NN	O	O
the	NN	O	O
transduction	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
ACH-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
treated	NN	O	O
with	NN	O	O
HOCl	NN	O	O
or	NN	O	O
H2O2	NN	O	O
released	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
supernatants	NN	O	O
.	NN	O	O

The	NN	O	O
importance	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
induction	NN	O	O
by	NN	O	O
HOCl	NN	O	O
or	NN	O	O
H2O2	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
:	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
nuclear	NN	O	O
appearance	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
was	NN	O	O
promoted	NN	O	O
in	NN	O	O
untreated	NN	O	B-cell_line
cells	NN	O	I-cell_line
;	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
synergism	NN	O	O
between	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
HOCl	NN	O	O
was	NN	O	O
detected	NN	O	O
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
HOCl	NN	O	O
should	NN	O	O
be	NN	O	O
considered	NN	O	O
as	NN	O	O
an	NN	O	O
oxidative	NN	O	O
species	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
a	NN	O	O
T-lymphocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
through	NN	O	O
a	NN	O	O
transduction	NN	O	O
mechanism	NN	O	O
involving	NN	O	O
ROS	NN	O	O
,	NN	O	O
and	NN	O	O
having	NN	O	O
a	NN	O	O
long-distance	NN	O	O
effect	NN	O	O
through	NN	O	O
subsequent	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
.	NN	O	O

-DOCSTART-	O

Rapid	NN	O	O
Ca2+-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
Rap1	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
platelets	NN	O	I-cell_type
.	NN	O	O

Rap1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
small	NN	O	O
,	NN	O	O
Ras-like	NN	O	B-protein
GTPase	NN	O	I-protein
whose	NN	O	O
function	NN	O	O
and	NN	O	O
regulation	NN	O	O
are	NN	O	O
still	NN	O	O
largely	NN	O	O
unknown	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
novel	NN	O	O
assay	NN	O	O
to	NN	O	O
monitor	NN	O	O
the	NN	O	O
active	NN	O	O
,	NN	O	O
GTP-bound	NN	O	B-protein
form	NN	O	I-protein
of	NN	O	O
Rap1	NN	O	B-protein
based	NN	O	O
on	NN	O	O
the	NN	O	O
differential	NN	O	O
affinity	NN	O	O
of	NN	O	O
Rap1GTP	NN	O	B-protein
and	NN	O	O
Rap1GDP	NN	O	B-protein
for	NN	O	O
the	NN	O	O
Rap	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
RalGDS	NN	O	B-protein
(	NN	O	O
RBD	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
blood	NN	O	B-cell_type
platelets	NN	O	I-cell_type
with	NN	O	O
alpha-thrombin	NN	O	B-protein
or	NN	O	O
other	NN	O	O
platelet	NN	O	O
activators	NN	O	O
caused	NN	O	O
a	NN	O	O
rapid	NN	O	O
and	NN	O	O
strong	NN	O	O
induction	NN	O	O
of	NN	O	O
Rap1	NN	O	B-protein
that	NN	O	O
associated	NN	O	O
with	NN	O	O
RBD	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Binding	NN	O	O
to	NN	O	O
RBD	NN	O	B-protein
increased	NN	O	O
from	NN	O	O
undetectable	NN	O	O
levels	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
platelets	NN	O	I-cell_type
to	NN	O	O
>	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
total	NN	O	O
Rap1	NN	O	B-protein
within	NN	O	O
30	NN	O	O
s	NN	O	O
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

An	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
concentration	NN	O	O
is	NN	O	O
both	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
Rap1	NN	O	B-protein
activation	NN	O	O
since	NN	O	O
it	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
agents	NN	O	O
that	NN	O	O
increase	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
and	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
Ca2+-chelating	NN	O	O
agent	NN	O	O
.	NN	O	O

Neither	NN	O	O
inhibition	NN	O	O
of	NN	O	O
translocation	NN	O	O
of	NN	O	O
Rap1	NN	O	B-protein
to	NN	O	O
the	NN	O	O
cytoskeleton	NN	O	O
nor	NN	O	O
inhibition	NN	O	O
of	NN	O	O
platelet	NN	O	O
aggregation	NN	O	O
affected	NN	O	O
thrombin	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
Rap1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
prostaglandin	NN	O	O
I2	NN	O	O
(	NN	O	O
PGI2	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
strong	NN	O	O
negative	NN	O	O
regulator	NN	O	O
of	NN	O	O
platelet	NN	O	B-cell_type
function	NN	O	O
,	NN	O	O
inhibited	NN	O	O
agonist-induced	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
Ca2+-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
Rap1	NN	O	B-protein
.	NN	O	O

From	NN	O	O
our	NN	O	O
results	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
Rap1	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
platelets	NN	O	B-cell_type
is	NN	O	O
an	NN	O	O
important	NN	O	O
common	NN	O	O
event	NN	O	O
in	NN	O	O
early	NN	O	O
agonist-induced	NN	O	O
signalling	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
activation	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
an	NN	O	O
increased	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
concentration	NN	O	O

-DOCSTART-	O

Physical	NN	O	O
interactions	NN	O	O
between	NN	O	O
Ets	NN	O	B-protein
and	NN	O	I-protein
NF-kappaB/NFAT	NN	O	I-protein
proteins	NN	O	I-protein
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
their	NN	O	O
cooperative	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
of	NN	O	O
many	NN	O	O
inducible	NN	O	B-DNA
T-cell	NN	O	I-DNA
genes	NN	O	I-DNA
contain	NN	O	O
adjacent	NN	O	O
or	NN	O	O
overlapping	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
and	NN	O	I-protein
NF-kappaB/NFAT	NN	O	I-protein
families	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Similar	NN	O	O
arrays	NN	O	O
of	NN	O	O
functionally	NN	O	O
important	NN	O	O
NF-kappaB/NFAT	NN	O	B-DNA
and	NN	O	I-DNA
Ets	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
enhancers	NN	O	I-DNA
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
viruses	NN	O	O
types	NN	O	O
1	NN	O	O
and	NN	O	O
2	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
HIV-2	NN	O	O
)	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
pattern	NN	O	O
of	NN	O	O
nuclear	NN	O	B-DNA
protein	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
reflects	NN	O	O
an	NN	O	O
evolutionarily	NN	O	O
conserved	NN	O	O
mechanism	NN	O	O
for	NN	O	O
regulating	NN	O	O
inducible	NN	O	O
T-cell	NN	O	O
gene	NN	O	O
expression	NN	O	O
that	NN	O	O
has	NN	O	O
been	NN	O	O
co-opted	NN	O	O
during	NN	O	O
HIV	NN	O	O
evolution	NN	O	O
.	NN	O	O

Despite	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
Ets	NN	O	B-protein
and	NN	O	I-protein
NF-kappaB/NFAT	NN	O	I-protein
proteins	NN	O	I-protein
cooperatively	NN	O	O
regulate	NN	O	O
inducible	NN	O	O
T-cell	NN	O	O
gene	NN	O	O
expression	NN	O	O
remained	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
studies	NN	O	O
described	NN	O	O
in	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
physical	NN	O	O
interaction	NN	O	O
between	NN	O	O
multiple	NN	O	O
Ets	NN	O	B-protein
and	NN	O	I-protein
NF-kappaB/NFAT	NN	O	I-protein
proteins	NN	O	I-protein
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
normal	NN	O	I-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
interaction	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
Ets	NN	O	B-DNA
domain	NN	O	I-DNA
of	NN	O	O
Ets	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
the	NN	O	O
C-terminal	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
Rel	NN	O	O
homology	NN	O	O
domains	NN	O	O
of	NN	O	O
NF-kappaB/NFAT	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
-	NN	O	O
NF-kappaB	NN	O	B-protein
/	NN	O	O
NFAT	NN	O	B-protein
interaction	NN	O	O
requires	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
both	NN	O	O
proteins	NN	O	O
,	NN	O	O
as	NN	O	O
it	NN	O	O
is	NN	O	O
abolished	NN	O	O
by	NN	O	O
the	NN	O	O
DNA	NN	O	O
intercalating	NN	O	O
agents	NN	O	O
propidium	NN	O	O
iodide	NN	O	O
and	NN	O	O
ethidium	NN	O	O
bromide	NN	O	O
and	NN	O	O
enhanced	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
synthetic	NN	O	O
oligonucleotides	NN	O	O
containing	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
Ets	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

A	NN	O	O
dominant-negative	NN	O	O
mutant	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
p50	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
DNA	NN	O	O
but	NN	O	O
fails	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
Ets	NN	O	B-protein
proteins	NN	O	I-protein
inhibits	NN	O	O
the	NN	O	O
synergistic	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
and	NN	O	I-DNA
HIV-2	NN	O	I-DNA
enhancers	NN	O	I-DNA
by	NN	O	O
NF-kappaB	NN	O	B-protein
(	NN	O	O
p50	NN	O	B-protein
+	NN	O	O
p65	NN	O	B-protein
)	NN	O	O
and	NN	O	O
Ets-1	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
physical	NN	O	O
interaction	NN	O	O
between	NN	O	O
Ets	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
proteins	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
and	NN	O	I-DNA
HIV-2	NN	O	I-DNA
enhancers	NN	O	I-DNA
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
evolutionarily	NN	O	O
conserved	NN	O	O
physical	NN	O	O
interactions	NN	O	O
between	NN	O	O
Ets	NN	O	B-protein
and	NN	O	I-protein
NF-kappaB/NFAT	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
important	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
T-cell	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
viruses	NN	O	O
.	NN	O	O

These	NN	O	O
interactions	NN	O	O
represent	NN	O	O
a	NN	O	O
potential	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
novel	NN	O	O
immunosuppressive	NN	O	O
and	NN	O	O
antiviral	NN	O	O
therapies	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
acute	NN	O	B-DNA
myeloid	NN	O	I-DNA
leukemia	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
AML1	NN	O	B-DNA
,	NN	O	O
regulates	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
and	NN	O	O
hemopoietic	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
antagonistically	NN	O	O
by	NN	O	O
two	NN	O	O
alternative	NN	O	O
spliced	NN	O	O
forms	NN	O	O
.	NN	O	O

The	NN	O	O
AML1	NN	O	B-DNA
gene	NN	O	I-DNA
on	NN	O	O
chromosome	NN	O	B-DNA
21	NN	O	I-DNA
is	NN	O	O
disrupted	NN	O	O
in	NN	O	O
the	NN	O	O
(	NN	O	B-DNA
8	NN	O	I-DNA
;	NN	O	I-DNA
21	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q22	NN	O	I-DNA
;	NN	O	I-DNA
q22	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	I-DNA
(	NN	O	I-DNA
3	NN	O	I-DNA
;	NN	O	I-DNA
21	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q26	NN	O	I-DNA
;	NN	O	I-DNA
q22	NN	O	I-DNA
)	NN	O	I-DNA
translocations	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
myelogenous	NN	O	O
leukemias	NN	O	O
and	NN	O	O
encodes	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

From	NN	O	O
AML1	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
two	NN	O	O
representative	NN	O	O
forms	NN	O	O
of	NN	O	O
proteins	NN	O	O
,	NN	O	O
AML1a	NN	O	B-protein
and	NN	O	O
AML1b	NN	O	B-protein
,	NN	O	O
are	NN	O	O
produced	NN	O	O
by	NN	O	O
an	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
.	NN	O	O

Both	NN	O	O
forms	NN	O	O
have	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
but	NN	O	O
AML1a	NN	O	B-protein
lacks	NN	O	O
a	NN	O	O
putative	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
domain	NN	O	O
which	NN	O	O
AML1b	NN	O	B-protein
has	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
AML1a	NN	O	B-protein
,	NN	O	O
which	NN	O	O
solely	NN	O	O
has	NN	O	O
no	NN	O	O
effects	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
regulator	NN	O	O
,	NN	O	O
dominantly	NN	O	O
suppresses	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
by	NN	O	O
AML1b	NN	O	B-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
AML1a	NN	O	B-protein
exhibits	NN	O	O
the	NN	O	O
higher	NN	O	O
affinity	NN	O	O
for	NN	O	O
DNA-binding	NN	O	O
than	NN	O	O
AML1b	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
a	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
form	NN	O	I-protein
of	NN	O	O
AML1	NN	O	B-protein
,	NN	O	O
AML1a	NN	O	B-protein
,	NN	O	O
totally	NN	O	O
suppressed	NN	O	O
granulocytic	NN	O	O
differentiation	NN	O	O
otherwise	NN	O	O
induced	NN	O	O
by	NN	O	O
granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
when	NN	O	O
AML1a	NN	O	B-protein
was	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
32Dc13	NN	O	B-cell_type
murine	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Such	NN	O	O
differentiation	NN	O	O
block	NN	O	O
by	NN	O	O
AML1a	NN	O	B-protein
was	NN	O	O
canceled	NN	O	O
by	NN	O	O
the	NN	O	O
concomitant	NN	O	O
overexpression	NN	O	O
of	NN	O	O
AML1b	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
form	NN	O	O
of	NN	O	O
AML1	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
an	NN	O	O
altered	NN	O	O
expression	NN	O	O
level	NN	O	O
of	NN	O	O
AML1	NN	O	B-protein
along	NN	O	O
with	NN	O	O
the	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
in	NN	O	O
several	NN	O	O
hemopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
these	NN	O	O
cases	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
,	NN	O	O
the	NN	O	O
AML1	NN	O	B-protein
expression	NN	O	O
level	NN	O	O
is	NN	O	O
a	NN	O	O
potential	NN	O	O
regulator	NN	O	O
for	NN	O	O
myeloid	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
potently	NN	O	O
up-regulates	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
of	NN	O	O
RANTES	NN	O	B-protein
,	NN	O	O
a	NN	O	O
chemokine	NN	O	O
that	NN	O	O
blocks	NN	O	O
HIV	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
complex	NN	O	O
network	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
and	NN	O	O
immunoregulatory	NN	O	O
responses	NN	O	O
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
HIV	NN	O	O
infection	NN	O	O
.	NN	O	O

RANTES	NN	O	B-protein
(	NN	O	O
regulated	NN	O	B-protein
upon	NN	O	I-protein
activation	NN	O	I-protein
,	NN	O	I-protein
normal	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
expressed	NN	O	I-protein
and	NN	O	I-protein
secreted	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
cytokine	NN	O	B-protein
that	NN	O	O
belongs	NN	O	O
to	NN	O	O
the	NN	O	O
beta-chemokine	NN	O	B-protein
family	NN	O	I-protein
;	NN	O	O
it	NN	O	O
is	NN	O	O
chemoattractant	NN	O	O
for	NN	O	O
CD4+/CD45RO	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
it	NN	O	O
is	NN	O	O
produced	NN	O	O
by	NN	O	O
various	NN	O	O
cell	NN	O	O
types	NN	O	O
including	NN	O	O
CD8+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
suppress	NN	O	O
replication	NN	O	O
of	NN	O	O
macrophage-tropic	NN	O	O
strains	NN	O	O
of	NN	O	O
HIV	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
RANTES	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
the	NN	O	O
RANTES	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
transient	NN	O	O
expression	NN	O	O
and	NN	O	O
gel-mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
:	NN	O	O
1	NN	O	O
)	NN	O	O
RANTES	NN	O	B-protein
promoter	NN	O	O
activity	NN	O	O
is	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
PMA	NN	O	O
plus	NN	O	O
ionomycin	NN	O	O
,	NN	O	O
coexpression	NN	O	O
of	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
the	NN	O	O
proinflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
TNF-alpha	NN	O	B-protein
and	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
costimulatory	NN	O	O
pathway	NN	O	O
;	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
RANTES	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
contains	NN	O	O
four	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
at	NN	O	O
positions	NN	O	B-DNA
-30	NN	O	I-DNA
,	NN	O	I-DNA
-44	NN	O	I-DNA
,	NN	O	I-DNA
-213	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
-579	NN	O	I-DNA
relative	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
;	NN	O	O
3	NN	O	O
)	NN	O	O
one	NN	O	O
site	NN	O	O
(	NN	O	O
-213	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
NF-AT	NN	O	B-DNA
(	NN	O	I-DNA
nuclear	NN	O	I-DNA
factor	NN	O	I-DNA
of	NN	O	I-DNA
activated	NN	O	I-DNA
T	NN	O	I-DNA
cells	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
that	NN	O	O
also	NN	O	O
has	NN	O	O
weak	NN	O	O
affinity	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
most	NN	O	O
distal	NN	O	O
site	NN	O	O
(	NN	O	O
-579	NN	O	O
)	NN	O	O
also	NN	O	O
serves	NN	O	O
as	NN	O	O
a	NN	O	O
CD28-responsive	NN	O	B-DNA
element	NN	O	I-DNA
;	NN	O	O
and	NN	O	O
4	NN	O	O
)	NN	O	O
mutation	NN	O	O
on	NN	O	O
any	NN	O	O
of	NN	O	O
those	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
or	NN	O	O
coexpression	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
cytoplasmic	NN	O	B-protein
inhibitor	NN	O	I-protein
of	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
potent	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
,	NN	O	O
is	NN	O	O
also	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
RANTES	NN	O	B-protein
,	NN	O	O
a	NN	O	O
chemokine	NN	O	B-protein
that	NN	O	O
blocks	NN	O	O
infection	NN	O	O
by	NN	O	O
macrophage-tropic	NN	O	O
strains	NN	O	O
of	NN	O	O
HIV	NN	O	O
.	NN	O	O

-DOCSTART-	O

Pivotal	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
NFIL3/E4BP4	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
interleukin	NN	O	B-protein
3	NN	O	I-protein
-mediated	NN	O	O
survival	NN	O	O
of	NN	O	O
pro-B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
E2A-HLF	NN	O	B-protein
(	NN	O	I-protein
hepatic	NN	O	I-protein
leukemia	NN	O	I-protein
factor	NN	O	I-protein
)	NN	O	I-protein
oncoprotein	NN	O	I-protein
,	NN	O	O
generated	NN	O	O
in	NN	O	O
pro-B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
fusion	NN	O	O
of	NN	O	O
the	NN	O	O
trans-activation	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
E2A	NN	O	B-protein
to	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
region/leucine	NN	O	I-protein
zipper	NN	O	I-protein
(	NN	O	I-protein
bZIP	NN	O	I-protein
)	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
HLF	NN	O	B-protein
,	NN	O	O
functions	NN	O	O
as	NN	O	O
an	NN	O	O
anti-apoptotic	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
leukemic	NN	O	O
cell	NN	O	O
transformation	NN	O	O
.	NN	O	O

When	NN	O	O
introduced	NN	O	O
into	NN	O	O
interleukin	NN	O	B-protein
3	NN	O	I-protein
(	NN	O	O
IL-3	NN	O	B-protein
)	NN	O	O
-dependent	NN	O	O
mouse	NN	O	O
pro-B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
E2A-HLF	NN	O	B-protein
prevents	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
growth	NN	O	B-protein
factor	NN	O	I-protein
deprivation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
IL-3	NN	O	B-protein
mediates	NN	O	O
cell	NN	O	O
survival	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
whose	NN	O	O
activity	NN	O	O
can	NN	O	O
be	NN	O	O
constitutively	NN	O	O
replaced	NN	O	O
by	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
oncoprotein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
considered	NN	O	O
four	NN	O	O
bZIP	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
as	NN	O	O
candidates	NN	O	O
for	NN	O	O
this	NN	O	O
putative	NN	O	O
IL-3-regulated	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
each	NN	O	O
of	NN	O	O
which	NN	O	O
binds	NN	O	O
avidly	NN	O	O
to	NN	O	O
the	NN	O	O
DNA	NN	O	B-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
recognized	NN	O	O
by	NN	O	O
E2A-HLF	NN	O	B-protein
and	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
Caenorhabditis	NN	O	B-protein
elegans	NN	O	I-protein
CES-2	NN	O	I-protein
(	NN	O	O
cell	NN	O	B-protein
death	NN	O	I-protein
specification	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
neuron-specific	NN	O	B-protein
mediator	NN	O	I-protein
of	NN	O	I-protein
cell	NN	O	I-protein
death	NN	O	I-protein
.	NN	O	O

The	NN	O	O
expression	NN	O	O
and	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
Nfil3	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
also	NN	O	O
called	NN	O	O
E4bp4	NN	O	B-protein
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
Hlf	NN	O	B-protein
,	NN	O	O
Dbp	NN	O	B-protein
,	NN	O	O
or	NN	O	O
Tef	NN	O	B-protein
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
IL-3	NN	O	B-protein
in	NN	O	O
mouse	NN	O	B-cell_line
pro-B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
Baf-3	NN	O	B-cell_line
and	NN	O	O
FL5.12	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
Nfil3/E4bp4	NN	O	B-protein
is	NN	O	O
regulated	NN	O	O
as	NN	O	O
a	NN	O	O
``	NN	O	B-DNA
delayed-early	NN	O	I-DNA
''	NN	O	I-DNA
IL-3-responsive	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
requiring	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
IL-3	NN	O	B-protein
,	NN	O	O
enforced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
NFIL3/E4BP4	NN	O	I-DNA
cDNA	NN	O	I-DNA
promoted	NN	O	O
the	NN	O	O
survival	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
IL-3-dependent	NN	O	B-cell_type
pro-B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
implicate	NN	O	O
NFIL3/E4BP4	NN	O	B-protein
(	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
regulated	NN	O	I-protein
by	NN	O	I-protein
IL-3/adenovirus	NN	O	I-protein
E4	NN	O	I-protein
promoter	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
in	NN	O	O
a	NN	O	O
distinct	NN	O	O
growth	NN	O	O
factor-regulated	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
that	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
early	NN	O	B-cell_type
B-cell	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
whose	NN	O	O
alteration	NN	O	O
by	NN	O	O
E2A-HLF	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
childhood	NN	O	O
B	NN	O	O
lineage	NN	O	O
leukemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

Detection	NN	O	O
of	NN	O	O
adenovirus	NN	O	O
DNA	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
assay	NN	O	O
.	NN	O	O

Adenovirus	NN	O	O
can	NN	O	O
establish	NN	O	O
persistent	NN	O	O
infections	NN	O	O
which	NN	O	O
may	NN	O	O
reactivate	NN	O	O
and	NN	O	O
cause	NN	O	O
disease	NN	O	O
in	NN	O	O
immunocompromised	NN	O	O
hosts	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
have	NN	O	O
been	NN	O	O
postulated	NN	O	O
to	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
site	NN	O	O
of	NN	O	O
adenoviral	NN	O	O
persistence	NN	O	O
based	NN	O	O
upon	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
isolate	NN	O	O
adenovirus	NN	O	O
from	NN	O	O
tonsils	NN	O	O
and	NN	O	O
to	NN	O	O
detect	NN	O	O
adenovirus	NN	O	O
DNA	NN	O	O
by	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
a	NN	O	O
more	NN	O	O
sensitive	NN	O	O
and	NN	O	O
specific	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
assay	NN	O	O
was	NN	O	O
developed	NN	O	O
to	NN	O	O
detect	NN	O	O
adenovirus	NN	O	O
DNA	NN	O	O
.	NN	O	O

Two	NN	O	O
sets	NN	O	O
of	NN	O	O
nested	NN	O	O
primers	NN	O	O
were	NN	O	O
designed	NN	O	O
to	NN	O	O
conserved	NN	O	O
sequences	NN	O	O
in	NN	O	O
the	NN	O	O
adenovirus	NN	O	B-DNA
E1A	NN	O	I-DNA
and	NN	O	I-DNA
hexon	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
E1A	NN	O	B-DNA
and	NN	O	I-DNA
hexon	NN	O	I-DNA
primers	NN	O	I-DNA
amplified	NN	O	O
DNA	NN	O	O
from	NN	O	O
representative	NN	O	O
adenoviral	NN	O	O
serotypes	NN	O	O
in	NN	O	O
all	NN	O	O
six	NN	O	O
adenoviral	NN	O	O
groups	NN	O	O
(	NN	O	O
A-F	NN	O	O
)	NN	O	O
.	NN	O	O

Both	NN	O	O
primers	NN	O	O
detected	NN	O	O
a	NN	O	O
single	NN	O	O
copy	NN	O	O
of	NN	O	O
the	NN	O	O
adenovirus	NN	O	B-DNA
type	NN	O	I-DNA
2	NN	O	I-DNA
genome	NN	O	I-DNA
but	NN	O	O
were	NN	O	O
less	NN	O	O
sensitive	NN	O	O
for	NN	O	O
the	NN	O	O
group	NN	O	O
B	NN	O	O
type	NN	O	O
35	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
33	NN	O	O
PBMC	NN	O	B-cell_type
specimens	NN	O	I-cell_type
from	NN	O	O
healthy	NN	O	O
adults	NN	O	O
and	NN	O	O
only	NN	O	O
one	NN	O	O
of	NN	O	O
40	NN	O	O
pediatric	NN	O	O
samples	NN	O	O
was	NN	O	O
positive	NN	O	O
(	NN	O	O
at	NN	O	O
a	NN	O	O
low	NN	O	O
level	NN	O	O
)	NN	O	O
for	NN	O	O
adenovirus	NN	O	O
DNA	NN	O	O
by	NN	O	O
nested	NN	O	O
PCR	NN	O	O
assay	NN	O	O
.	NN	O	O

In	NN	O	O
comparison	NN	O	O
,	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
two	NN	O	O
children	NN	O	O
with	NN	O	O
fatal	NN	O	O
adenoviral	NN	O	O
infection	NN	O	O
were	NN	O	O
both	NN	O	O
strongly	NN	O	O
positive	NN	O	O
for	NN	O	O
adenovirus	NN	O	O
DNA	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
a	NN	O	O
previous	NN	O	O
study	NN	O	O
,	NN	O	O
PBMC	NN	O	B-cell_type
are	NN	O	O
not	NN	O	O
a	NN	O	O
common	NN	O	O
site	NN	O	O
of	NN	O	O
persistent	NN	O	O
group	NN	O	O
C	NN	O	O
adenoviral	NN	O	O
infection	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
assay	NN	O	O
of	NN	O	O
PBMC	NN	O	B-cell_type
by	NN	O	O
the	NN	O	O
adenovirus-specific	NN	O	O
PCR	NN	O	O
may	NN	O	O
help	NN	O	O
detect	NN	O	O
early	NN	O	O
invasive	NN	O	O
disease	NN	O	O
and	NN	O	O
warrants	NN	O	O
further	NN	O	O
evaluation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Common	NN	O	O
and	NN	O	O
distinct	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
utilized	NN	O	O
by	NN	O	O
platelet	NN	O	B-protein
activating	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
FMLP	NN	O	B-protein
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
with	NN	O	O
chemoattractants	NN	O	B-protein
FMLP	NN	O	B-protein
or	NN	O	O
platelet	NN	O	B-protein
activating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
PAF	NN	O	B-protein
)	NN	O	O
results	NN	O	O
in	NN	O	O
different	NN	O	O
but	NN	O	O
overlapping	NN	O	O
functional	NN	O	O
responses	NN	O	O
.	NN	O	O

We	NN	O	O
questioned	NN	O	O
whether	NN	O	O
these	NN	O	O
differences	NN	O	O
might	NN	O	O
reflect	NN	O	O
patterns	NN	O	O
of	NN	O	O
intracellular	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
with	NN	O	O
either	NN	O	O
PAF	NN	O	B-protein
or	NN	O	O
FMLP	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
equivalent	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
MAPk	NN	O	I-protein
)	NN	O	I-protein
homologue	NN	O	I-protein
38-kD	NN	O	B-protein
murine	NN	O	I-protein
MAP	NN	O	I-protein
kinase	NN	O	I-protein
homologous	NN	O	O
to	NN	O	O
HOG-1	NN	O	B-protein
(	NN	O	I-protein
p38	NN	O	I-protein
)	NN	O	I-protein
MAPk	NN	O	I-protein
.	NN	O	O

Neither	NN	O	O
FMLP	NN	O	B-protein
nor	NN	O	O
PAF	NN	O	B-protein
activated	NN	O	O
c-jun	NN	O	B-protein
NH2-terminal	NN	O	I-protein
MAPk	NN	O	I-protein
(	NN	O	O
JNKs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Under	NN	O	O
identical	NN	O	O
conditions	NN	O	O
,	NN	O	O
FMLP	NN	O	B-protein
but	NN	O	O
not	NN	O	O
PAF	NN	O	B-protein
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
significant	NN	O	O
p42/44	NN	O	B-protein
(	NN	O	I-protein
ERK	NN	O	I-protein
)	NN	O	I-protein
MAPk	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Both	NN	O	O
FMLP	NN	O	B-protein
and	NN	O	O
PAF	NN	O	B-protein
activated	NN	O	I-protein
MAP	NN	O	I-protein
kinase	NN	O	I-protein
kinase-3	NN	O	I-protein
(	NN	O	O
MKK3	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
known	NN	O	O
activator	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
.	NN	O	O

Both	NN	O	O
MAP	NN	O	B-protein
ERK	NN	O	I-protein
kinase	NN	O	I-protein
kinase-1	NN	O	I-protein
(	NN	O	O
MEKK1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
Raf	NN	O	B-protein
are	NN	O	O
activated	NN	O	O
strongly	NN	O	O
by	NN	O	O
FMLP	NN	O	B-protein
,	NN	O	O
but	NN	O	O
minimally	NN	O	O
by	NN	O	O
PAF	NN	O	B-protein
.	NN	O	O

Pertussis	NN	O	B-protein
toxin	NN	O	I-protein
blocked	NN	O	O
FMLP	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
p42/44	NN	O	B-protein
(	NN	O	I-protein
ERK	NN	O	I-protein
)	NN	O	I-protein
MAPk	NN	O	I-protein
cascade	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
that	NN	O	O
of	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
.	NN	O	O

A	NN	O	O
specific	NN	O	O
p38	NN	O	B-protein
MAPk	NN	O	I-protein
inhibitor	NN	O	O
(	NN	O	O
SK	NN	O	O
&	NN	O	O
F	NN	O	O
86002	NN	O	O
)	NN	O	O
blocked	NN	O	O
superoxide	NN	O	O
anion	NN	O	O
production	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
FMLP	NN	O	B-protein
and	NN	O	O
reduced	NN	O	O
adhesion	NN	O	O
and	NN	O	O
chemotaxis	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
PAF	NN	O	B-protein
or	NN	O	O
FMLP	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
distinct	NN	O	O
patterns	NN	O	O
of	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
for	NN	O	O
two	NN	O	O
chemoattractants	NN	O	B-protein
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
selective	NN	O	O
activation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
cascades	NN	O	O
may	NN	O	O
underlie	NN	O	O
different	NN	O	O
patterns	NN	O	O
of	NN	O	O
functional	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
of	NN	O	O
the	NN	O	O
novel	NN	O	B-protein
human	NN	O	I-protein
myeloid-cell-specific	NN	O	I-protein
C/EBP-epsilon	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Chicken	NN	O	B-protein
NF-M	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
in	NN	O	O
cooperation	NN	O	O
with	NN	O	O
either	NN	O	O
c-Myb	NN	O	B-protein
or	NN	O	O
v-Myb	NN	O	B-protein
,	NN	O	O
is	NN	O	O
active	NN	O	O
in	NN	O	O
the	NN	O	O
combinatorial	NN	O	O
activation	NN	O	O
of	NN	O	O
myeloid-cell-specific	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
heterologous	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
,	NN	O	O
such	NN	O	O
as	NN	O	O
embryonic	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
humans	NN	O	O
,	NN	O	O
similar	NN	O	O
effects	NN	O	O
were	NN	O	O
observed	NN	O	O
with	NN	O	O
homologous	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
CCAAT/enhancer-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
C/EBP	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcriptional	NN	O	B-protein
regulators	NN	O	I-protein
,	NN	O	O
especially	NN	O	O
the	NN	O	O
human	NN	O	O
homolog	NN	O	O
of	NN	O	O
chicken	NN	O	B-protein
NF-M	NN	O	I-protein
,	NN	O	O
C/EBP-beta	NN	O	B-protein
(	NN	O	O
NF-IL6	NN	O	B-protein
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
NF-IL6	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
nonmyeloid	NN	O	O
cell	NN	O	O
types	NN	O	O
and	NN	O	O
is	NN	O	O
strongly	NN	O	O
inducible	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
inflammatory	NN	O	O
stimuli	NN	O	O
,	NN	O	O
making	NN	O	O
it	NN	O	O
an	NN	O	O
unlikely	NN	O	O
candidate	NN	O	O
to	NN	O	O
have	NN	O	O
an	NN	O	O
exclusive	NN	O	O
role	NN	O	O
as	NN	O	O
a	NN	O	O
combinatorial	NN	O	O
differentiation	NN	O	O
switch	NN	O	O
during	NN	O	O
myelopoiesis	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

By	NN	O	O
using	NN	O	O
a	NN	O	O
reverse	NN	O	O
transcription-PCR-based	NN	O	O
approach	NN	O	O
and	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
primers	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-DNA
domains	NN	O	I-DNA
of	NN	O	O
highly	NN	O	O
homologous	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
C/EBP	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcriptional	NN	O	B-protein
regulators	NN	O	I-protein
,	NN	O	O
we	NN	O	O
have	NN	O	O
cloned	NN	O	O
a	NN	O	O
novel	NN	O	B-DNA
human	NN	O	I-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
C/EBP	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
,	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
human	NN	O	O
homolog	NN	O	O
of	NN	O	O
CRP1	NN	O	B-protein
,	NN	O	O
C/EBP-epsilon	NN	O	B-protein
.	NN	O	O

A	NN	O	O
1.2-kb	NN	O	B-DNA
cDNA	NN	O	I-DNA
encoding	NN	O	O
full-length	NN	O	O
human	NN	O	B-DNA
C/EBP-epsilon	NN	O	I-DNA
was	NN	O	O
cloned	NN	O	O
from	NN	O	O
a	NN	O	O
promyelocyte-late	NN	O	B-DNA
myeloblast-derived	NN	O	I-DNA
lambda	NN	O	I-DNA
gt11	NN	O	I-DNA
library	NN	O	I-DNA
.	NN	O	O

Molecular	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
cDNA	NN	O	O
and	NN	O	O
genomic	NN	O	O
clones	NN	O	O
indicated	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
two	NN	O	O
exons	NN	O	B-DNA
encoding	NN	O	O
a	NN	O	O
protein	NN	O	O
with	NN	O	O
an	NN	O	O
apparent	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
32	NN	O	O
kDa	NN	O	O
and	NN	O	O
a	NN	O	O
pI	NN	O	O
of	NN	O	O
9.5	NN	O	O
.	NN	O	O

Primer	NN	O	O
extension	NN	O	O
analysis	NN	O	O
of	NN	O	O
C/EBP-epsilon	NN	O	B-RNA
mRNA	NN	O	I-RNA
detected	NN	O	O
a	NN	O	O
single	NN	O	O
major	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
approximately	NN	O	O
200	NN	O	O
bp	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
start	NN	O	B-DNA
codon	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
putative	NN	O	O
promoter	NN	O	O
area	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
several	NN	O	O
other	NN	O	O
myeloid-cell-specific	NN	O	O
genes	NN	O	O
in	NN	O	O
that	NN	O	O
it	NN	O	O
contains	NN	O	O
no	NN	O	O
TATAAA	NN	O	B-DNA
box	NN	O	I-DNA
but	NN	O	O
has	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
purine-rich	NN	O	B-DNA
stretches	NN	O	I-DNA
with	NN	O	O
multiple	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
factors	NN	O	O
of	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcriptional	NN	O	B-protein
regulators	NN	O	I-protein
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analyses	NN	O	O
indicated	NN	O	O
a	NN	O	O
highly	NN	O	O
restricted	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
pattern	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
strongest	NN	O	O
expression	NN	O	O
occurring	NN	O	O
in	NN	O	O
promyelocyte	NN	O	B-cell_line
and	NN	O	I-cell_line
late-myeloblast-like	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Western	NN	O	O
blot	NN	O	O
and	NN	O	O
immunoprecipitation	NN	O	O
studies	NN	O	O
using	NN	O	O
rabbit	NN	O	B-protein
anti-C/EBP-epsilon	NN	O	I-protein
antibodies	NN	O	I-protein
raised	NN	O	O
against	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
portion	NN	O	I-protein
of	NN	O	O
C/EBP-epsilon	NN	O	B-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
1	NN	O	I-protein
to	NN	O	I-protein
115	NN	O	I-protein
)	NN	O	O
showed	NN	O	O
that	NN	O	O
C/EBP-epsilon	NN	O	B-protein
is	NN	O	O
a	NN	O	O
32-kDa	NN	O	B-protein
nuclear	NN	O	I-protein
phosphoprotein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
human	NN	O	O
C/EBP-epsilon	NN	O	B-protein
protein	NN	O	O
exhibited	NN	O	O
strong	NN	O	O
and	NN	O	O
specific	NN	O	O
binding	NN	O	O
to	NN	O	O
double-stranded	NN	O	B-DNA
DNA	NN	O	I-DNA
containing	NN	O	I-DNA
consensus	NN	O	I-DNA
C/EBP	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
the	NN	O	O
C/EBP-epsilon	NN	O	B-protein
sense	NN	O	O
and	NN	O	O
antisense	NN	O	O
expression	NN	O	O
constructs	NN	O	O
together	NN	O	O
with	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
reporter	NN	O	I-DNA
vectors	NN	O	I-DNA
containing	NN	O	O
myeloid-cell-specific	NN	O	B-DNA
c-mim	NN	O	I-DNA
and	NN	O	I-DNA
human	NN	O	I-DNA
myeloperoxidase	NN	O	I-DNA
promoters	NN	O	I-DNA
suggested	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
C/EBP-epsilon	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
myeloid-cell-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Transient	NN	O	O
tranfection	NN	O	O
of	NN	O	O
a	NN	O	O
promyelocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
NB4	NN	O	B-cell_line
)	NN	O	O
with	NN	O	O
a	NN	O	O
C/EBP-epsilon	NN	O	B-DNA
expression	NN	O	I-DNA
plasmid	NN	O	I-DNA
increased	NN	O	O
cell	NN	O	O
growth	NN	O	O
by	NN	O	O
sevenfold	NN	O	O
,	NN	O	O
while	NN	O	O
antisense	NN	O	O
C/EBP-epsilon	NN	O	B-protein
caused	NN	O	O
a	NN	O	O
fivefold	NN	O	O
decrease	NN	O	O
in	NN	O	O
clonal	NN	O	O
growth	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
tumour	NN	O	B-protein
associated	NN	O	I-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
antigen	NN	O	I-protein
A6H	NN	O	B-protein
is	NN	O	O
costimulatory	NN	O	O
for	NN	O	O
human	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
but	NN	O	I-cell_type
not	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
A6H	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
mAb	NN	O	O
)	NN	O	O
recognizes	NN	O	O
a	NN	O	O
120	NN	O	B-protein
,	NN	O	I-protein
000-140	NN	O	I-protein
,	NN	O	I-protein
000	NN	O	I-protein
MW	NN	O	I-protein
antigen	NN	O	I-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
similar	NN	O	O
densities	NN	O	O
on	NN	O	O
85-90	NN	O	O
%	NN	O	O
of	NN	O	O
human	NN	O	O
CD4+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
on	NN	O	O
renal	NN	O	O
cell	NN	O	O
carcinomas	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
A6H	NN	O	B-protein
mAb	NN	O	I-protein
induced	NN	O	O
a	NN	O	O
costimulatory	NN	O	O
signal	NN	O	O
in	NN	O	O
anti-CD3	NN	O	B-cell_line
activated	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
A6H	NN	O	B-protein
costimulated	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
in	NN	O	O
purified	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
the	NN	O	O
CD8+	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
subpopulation	NN	O	I-cell_type
failed	NN	O	O
to	NN	O	O
respond	NN	O	O
.	NN	O	O

CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
costimulated	NN	O	O
with	NN	O	O
the	NN	O	O
A6H	NN	O	B-protein
mAb	NN	O	I-protein
upregulated	NN	O	O
CD80	NN	O	B-protein
,	NN	O	O
CD86	NN	O	B-protein
,	NN	O	O
CD71	NN	O	B-protein
,	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	I-protein
IL-2	NN	O	I-protein
)	NN	O	I-protein
R	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
IL-2R	NN	O	B-protein
gamma	NN	O	I-protein
,	NN	O	O
while	NN	O	O
no	NN	O	O
corresponding	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
these	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
molecules	NN	O	I-protein
was	NN	O	O
seen	NN	O	O
in	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
A6H	NN	O	B-protein
mAb	NN	O	I-protein
costimulus	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
OCT-1	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
which	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
transcriptional	NN	O	B-protein
regulators	NN	O	I-protein
of	NN	O	O
several	NN	O	O
cytokine	NN	O	B-protein
and	NN	O	O
cytokine	NN	O	B-protein
receptor	NN	O	O
genes	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
and	NN	O	I-DNA
IL-2R	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Co-ligation	NN	O	O
of	NN	O	O
the	NN	O	O
A6H	NN	O	B-protein
antigen	NN	O	O
and	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
complex	NN	O	I-protein
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
in	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
no	NN	O	O
increase	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
octamer-binding	NN	O	B-protein
(	NN	O	I-protein
Oct	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
was	NN	O	O
seen	NN	O	O
compared	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
no	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
was	NN	O	O
seen	NN	O	O
in	NN	O	O
A6H	NN	O	B-protein
costimulated	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggests	NN	O	O
that	NN	O	O
both	NN	O	O
proximal	NN	O	O
steps	NN	O	O
in	NN	O	O
CD8+	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
later	NN	O	O
phases	NN	O	O
are	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
A6H	NN	O	B-protein
ligation	NN	O	O
.	NN	O	O

Molecular	NN	O	O
differences	NN	O	O
of	NN	O	O
the	NN	O	O
A6H	NN	O	B-protein
molecule	NN	O	I-protein
or	NN	O	O
distinct	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
A6H	NN	O	B-protein
transduced	NN	O	O
AP-1	NN	O	B-protein
activation	NN	O	O
pathway	NN	O	O
may	NN	O	O
exist	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
subpopulations	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
from	NN	O	O
the	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
(	NN	O	O
Saimiri	NN	O	O
boliviensis	NN	O	O
boliviensis	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
glucocorticoid-resistant	NN	O	O
primate	NN	O	O
.	NN	O	O

New	NN	O	O
World	NN	O	O
primates	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
have	NN	O	O
elevated	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
apparent	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
squirrel	NN	O	B-cell_type
monkey	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
5-fold	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
apparent	NN	O	O
Kd	NN	O	O
,	NN	O	O
20.9	NN	O	O
+/-	NN	O	O
1.8	NN	O	O
and	NN	O	O
4.3	NN	O	O
+/-	NN	O	O
0.2	NN	O	O
nmol/L	NN	O	O
,	NN	O	O
respectively	NN	O	O
;	NN	O	O
n	NN	O	O
=	NN	O	O
3	NN	O	O
)	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
previous	NN	O	O
studies	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
two	NN	O	O
species	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
first	NN	O	O
step	NN	O	O
in	NN	O	O
understanding	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
decreased	NN	O	O
binding	NN	O	O
affinity	NN	O	O
in	NN	O	O
New	NN	O	O
World	NN	O	O
primates	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
reverse	NN	O	O
transcription-PCR	NN	O	O
to	NN	O	O
clone	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
from	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
liver	NN	O	O
and	NN	O	O
have	NN	O	O
compared	NN	O	O
the	NN	O	O
sequence	NN	O	O
to	NN	O	O
receptor	NN	O	O
sequences	NN	O	O
obtained	NN	O	O
from	NN	O	O
owl	NN	O	O
monkey	NN	O	O
liver	NN	O	O
,	NN	O	O
cotton-top	NN	O	B-cell_line
tamarin	NN	O	I-cell_line
B95-8	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
approximately	NN	O	O
97	NN	O	O
%	NN	O	O
identical	NN	O	O
in	NN	O	O
nucleotide	NN	O	O
and	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
ligand-binding	NN	O	B-protein
domain	NN	O	I-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
528-777	NN	O	I-protein
)	NN	O	O
of	NN	O	O
the	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
contains	NN	O	O
four	NN	O	O
amino	NN	O	O
acid	NN	O	O
differences	NN	O	O
(	NN	O	O
Ser551	NN	O	O
to	NN	O	O
Thr	NN	O	O
,	NN	O	O
Ser616	NN	O	O
to	NN	O	O
Ala	NN	O	O
,	NN	O	O
Ala618	NN	O	O
to	NN	O	O
Ser	NN	O	O
,	NN	O	O
and	NN	O	O
Ile761	NN	O	O
to	NN	O	O
Leu	NN	O	O
)	NN	O	O
,	NN	O	O
all	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
owl	NN	O	B-protein
monkey	NN	O	I-protein
and	NN	O	I-protein
cotton-top	NN	O	I-protein
tamarin	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
421-486	NN	O	I-protein
)	NN	O	O
is	NN	O	O
completely	NN	O	O
conserved	NN	O	O
among	NN	O	O
human	NN	O	B-protein
,	NN	O	I-protein
squirrel	NN	O	I-protein
monkey	NN	O	I-protein
,	NN	O	I-protein
owl	NN	O	I-protein
monkey	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
cotton-top	NN	O	I-protein
tamarin	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Twenty-two	NN	O	O
differences	NN	O	O
from	NN	O	O
the	NN	O	O
human	NN	O	O
sequence	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
region	NN	O	I-protein
(	NN	O	O
amino	NN	O	B-protein
acids	NN	O	I-protein
1-421	NN	O	I-protein
)	NN	O	O
of	NN	O	O
the	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
receptor	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
substitutions	NN	O	O
in	NN	O	O
the	NN	O	O
ligand-binding	NN	O	B-protein
domain	NN	O	I-protein
matched	NN	O	O
mutations	NN	O	O
known	NN	O	O
to	NN	O	O
influence	NN	O	O
binding	NN	O	O
affinity	NN	O	O
in	NN	O	O
other	NN	O	O
species	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
substitutions	NN	O	O
per	NN	O	O
se	NN	O	O
were	NN	O	O
responsible	NN	O	O
for	NN	O	O
decreased	NN	O	O
affinity	NN	O	O
,	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
and	NN	O	O
human	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
were	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
TNT	NN	O	O
Coupled	NN	O	O
Reticulocyte	NN	O	O
Lysate	NN	O	O
System	NN	O	O
.	NN	O	O

Expressions	NN	O	O
of	NN	O	O
human	NN	O	O
and	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
a	NN	O	O
squirrel	NN	O	B-protein
monkey	NN	O	I-protein
receptor	NN	O	I-protein
in	NN	O	O
which	NN	O	O
Phe774	NN	O	O
was	NN	O	O
mutated	NN	O	O
to	NN	O	O
Leu	NN	O	O
(	NN	O	O
F774L	NN	O	O
)	NN	O	O
were	NN	O	O
similar	NN	O	O
.	NN	O	O

When	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
TNT	NN	O	O
System	NN	O	O
,	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
and	NN	O	O
human	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
had	NN	O	O
similar	NN	O	O
,	NN	O	O
high	NN	O	O
affinity	NN	O	O
binding	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
apparent	NN	O	O
Kd	NN	O	O
,	NN	O	O
5.9	NN	O	O
+/-	NN	O	O
1.2	NN	O	O
and	NN	O	O
4.3	NN	O	O
+/-	NN	O	O
0.5	NN	O	O
nmol/L	NN	O	O
,	NN	O	O
respectively	NN	O	O
;	NN	O	O
n	NN	O	O
=	NN	O	O
3	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
squirrel	NN	O	B-protein
monkey	NN	O	I-protein
F774L	NN	O	I-protein
receptor	NN	O	I-protein
had	NN	O	O
lower	NN	O	O
affinity	NN	O	O
binding	NN	O	O
(	NN	O	O
apparent	NN	O	O
Kd	NN	O	O
,	NN	O	O
20.4	NN	O	O
+/-	NN	O	O
2.0	NN	O	O
nmol/L	NN	O	O
;	NN	O	O
n	NN	O	O
=	NN	O	O
3	NN	O	O
)	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
substitutions	NN	O	O
within	NN	O	O
the	NN	O	O
ligand-binding	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
squirrel	NN	O	O
monkey	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
can	NN	O	O
not	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
decreased	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
these	NN	O	O
receptors	NN	O	O
in	NN	O	O
squirrel	NN	O	B-cell_type
monkey	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
affinity	NN	O	O
is	NN	O	O
probably	NN	O	O
influenced	NN	O	O
by	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cytosolic	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
affect	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

Shared	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
within	NN	O	O
the	NN	O	O
human	NN	O	B-protein
interleukin-7	NN	O	I-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

A	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
X-linked	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
.	NN	O	O

Genetic	NN	O	O
evidence	NN	O	O
suggests	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
subunit	NN	O	I-protein
cause	NN	O	O
X-linked	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
(	NN	O	O
X-SCID	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
can	NN	O	O
be	NN	O	O
employed	NN	O	O
in	NN	O	O
receptor	NN	O	B-protein
complexes	NN	O	I-protein
for	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	I-protein
-4	NN	O	I-protein
,	NN	O	I-protein
-7	NN	O	I-protein
,	NN	O	I-protein
-9	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
-15	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
multiple	NN	O	O
signaling	NN	O	O
defects	NN	O	O
that	NN	O	O
would	NN	O	O
result	NN	O	O
from	NN	O	O
a	NN	O	O
defective	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
chain	NN	O	I-protein
in	NN	O	O
these	NN	O	O
receptors	NN	O	O
are	NN	O	O
proposed	NN	O	O
to	NN	O	O
cause	NN	O	O
the	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
of	NN	O	O
X-SCID	NN	O	O
patients	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
gene	NN	O	O
disruption	NN	O	O
of	NN	O	O
either	NN	O	O
IL-7	NN	O	B-protein
or	NN	O	O
the	NN	O	O
IL-7	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
IL-7R	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
subunit	NN	O	I-protein
in	NN	O	O
mice	NN	O	O
leads	NN	O	O
to	NN	O	O
immunological	NN	O	O
defects	NN	O	O
that	NN	O	O
are	NN	O	O
similar	NN	O	O
to	NN	O	O
human	NN	O	O
X-SCID	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
the	NN	O	O
functional	NN	O	O
importance	NN	O	O
of	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
in	NN	O	O
the	NN	O	O
IL-7R	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
structure/function	NN	O	O
analyses	NN	O	O
of	NN	O	O
the	NN	O	O
IL-7R	NN	O	B-protein
complex	NN	O	I-protein
using	NN	O	O
a	NN	O	O
chimeric	NN	O	B-protein
receptor	NN	O	I-protein
system	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
indeed	NN	O	O
critical	NN	O	O
for	NN	O	O
IL-7R	NN	O	B-protein
function	NN	O	O
.	NN	O	O

Nonetheless	NN	O	O
,	NN	O	O
only	NN	O	O
a	NN	O	O
limited	NN	O	O
portion	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
IL-7R	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
replacement	NN	O	O
of	NN	O	O
the	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
by	NN	O	O
a	NN	O	O
severely	NN	O	B-protein
truncated	NN	O	I-protein
erythropoeitin	NN	O	I-protein
receptor	NN	O	I-protein
does	NN	O	O
not	NN	O	O
affect	NN	O	O
measured	NN	O	O
IL-7R	NN	O	B-protein
signaling	NN	O	O
events	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
support	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
serves	NN	O	O
primarily	NN	O	O
to	NN	O	O
activate	NN	O	O
signal	NN	O	O
transduction	NN	O	O
by	NN	O	O
the	NN	O	O
IL-7R	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
while	NN	O	O
IL-7R	NN	O	B-protein
alpha	NN	O	I-protein
determines	NN	O	O
specific	NN	O	O
signaling	NN	O	O
events	NN	O	O
through	NN	O	O
its	NN	O	O
association	NN	O	O
with	NN	O	O
cytoplasmic	NN	O	B-protein
signaling	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
these	NN	O	O
studies	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
the	NN	O	O
molecular	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
X-SCID	NN	O	O
is	NN	O	O
due	NN	O	O
primarily	NN	O	O
to	NN	O	O
gamma	NN	O	B-protein
(	NN	O	I-protein
c	NN	O	I-protein
)	NN	O	I-protein
-mediated	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
IL-7	NN	O	B-protein
/IL-7R	NN	O	B-protein
system	NN	O	O

-DOCSTART-	O

S-allyl	NN	O	O
cysteine	NN	O	O
inhibits	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
leading	NN	O	O
to	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
activation	NN	O	O
which	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

We	NN	O	O
recently	NN	O	O
reported	NN	O	O
that	NN	O	O
a	NN	O	O
garlic	NN	O	O
compound	NN	O	O
,	NN	O	O
S-allyl	NN	O	O
cysteine	NN	O	O
(	NN	O	O
SAC	NN	O	O
)	NN	O	O
,	NN	O	O
protects	NN	O	O
bovine	NN	O	B-cell_type
pulmonary	NN	O	I-cell_type
artery	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
oxidant	NN	O	O
injury	NN	O	O
induced	NN	O	O
by	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
(	NN	O	O
H2O2	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
determined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
SAC	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
induced	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
and	NN	O	O
H2O2	NN	O	O
.	NN	O	O

Activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
an	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
using	NN	O	O
32P-labeled	NN	O	B-DNA
probe	NN	O	I-DNA
.	NN	O	O

SAC	NN	O	O
consistently	NN	O	O
exhibited	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
both	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
H2O2	NN	O	O
.	NN	O	O

Supershift	NN	O	O
with	NN	O	O
specific	NN	O	O
antibodies	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
subunits	NN	O	I-protein
confirmed	NN	O	O
that	NN	O	O
the	NN	O	O
inducible	NN	O	O
retarded	NN	O	O
bands	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
EMSA	NN	O	O
and	NN	O	O
p65-p50	NN	O	B-protein
heterodimer	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
SAC	NN	O	O
may	NN	O	O
act	NN	O	O
via	NN	O	O
antioxidant	NN	O	O
mechanisms	NN	O	O
to	NN	O	O
block	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Elf-2	NN	O	B-protein
,	NN	O	O
a	NN	O	O
rhombotin-2	NN	O	B-protein
binding	NN	O	I-protein
ets	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
:	NN	O	O
discovery	NN	O	O
and	NN	O	O
potential	NN	O	O
role	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Rhombotin-2	NN	O	B-DNA
(	NN	O	O
RBTN-2	NN	O	B-DNA
)	NN	O	O
is	NN	O	O
a	NN	O	O
proto-oncogene	NN	O	B-DNA
only	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
postulated	NN	O	O
that	NN	O	O
the	NN	O	O
oncogenic	NN	O	O
effect	NN	O	O
of	NN	O	O
RBTN-2	NN	O	B-protein
in	NN	O	O
T	NN	O	O
cells	NN	O	O
is	NN	O	O
likely	NN	O	O
mediated	NN	O	O
by	NN	O	O
binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
with	NN	O	O
T	NN	O	O
cell-specific	NN	O	O
expression	NN	O	O
.	NN	O	O

By	NN	O	O
screening	NN	O	O
a	NN	O	O
T	NN	O	O
cell	NN	O	O
cDNA	NN	O	B-DNA
library	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
ets	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
RBTN-2	NN	O	B-protein
.	NN	O	O

This	NN	O	O
protein	NN	O	O
was	NN	O	O
named	NN	O	O
elf-2	NN	O	B-protein
because	NN	O	O
its	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
is	NN	O	O
virtually	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
ets	NN	O	B-protein
family	NN	O	I-protein
member	NN	O	I-protein
elf-1	NN	O	B-protein
.	NN	O	O

Northern	NN	O	O
analyses	NN	O	O
showed	NN	O	O
similar	NN	O	O
levels	NN	O	O
of	NN	O	O
two	NN	O	O
elf-2	NN	O	B-RNA
transcripts	NN	O	I-RNA
(	NN	O	O
3.5	NN	O	O
kb	NN	O	O
and	NN	O	O
3.8	NN	O	O
kb	NN	O	O
)	NN	O	O
in	NN	O	O
all	NN	O	O
tissues	NN	O	O
except	NN	O	O
thymus	NN	O	O
.	NN	O	O

Thymocytes	NN	O	B-cell_type
expressed	NN	O	O
four-	NN	O	O
to	NN	O	O
10-fold	NN	O	O
greater	NN	O	O
amounts	NN	O	O
of	NN	O	O
the	NN	O	O
3.5	NN	O	B-RNA
kb	NN	O	I-RNA
transcript	NN	O	I-RNA
than	NN	O	O
other	NN	O	O
tissues	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analyses	NN	O	O
of	NN	O	O
cDNA	NN	O	B-DNA
clones	NN	O	I-DNA
indicated	NN	O	O
that	NN	O	O
these	NN	O	O
transcripts	NN	O	O
encode	NN	O	O
proteins	NN	O	O
differing	NN	O	O
only	NN	O	O
at	NN	O	O
their	NN	O	O
amino	NN	O	B-protein
termini	NN	O	I-protein
,	NN	O	O
and	NN	O	O
likely	NN	O	O
represent	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
isoforms	NN	O	O
.	NN	O	O

These	NN	O	O
isoforms	NN	O	O
(	NN	O	O
elf-2a	NN	O	B-protein
and	NN	O	O
elf-2b	NN	O	B-protein
)	NN	O	O
contain	NN	O	O
identical	NN	O	O
RBTN-2	NN	O	B-protein
binding	NN	O	I-protein
regions	NN	O	I-protein
and	NN	O	O
DNA-binding	NN	O	B-protein
domains	NN	O	I-protein
.	NN	O	O

Elf-2b	NN	O	B-protein
lacks	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
transactivation	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

The	NN	O	O
expression	NN	O	O
patterns	NN	O	O
suggest	NN	O	O
that	NN	O	O
RBTN-2	NN	O	B-protein
normally	NN	O	O
interacts	NN	O	O
equally	NN	O	O
with	NN	O	O
elf-2a	NN	O	B-protein
and	NN	O	O
elf-2b	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
when	NN	O	O
RBTN-2	NN	O	B-protein
is	NN	O	O
inappropriately	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
RBTN-2	NN	O	B-protein
would	NN	O	O
interact	NN	O	O
predominantly	NN	O	O
with	NN	O	O
elf-2b	NN	O	B-protein
;	NN	O	O
this	NN	O	O
interaction	NN	O	O
may	NN	O	O
lead	NN	O	O
to	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
effects	NN	O	O
on	NN	O	O
chromosome	NN	O	O
constitution	NN	O	O
of	NN	O	O
cell	NN	O	B-cell_line
hybrids	NN	O	I-cell_line
.	NN	O	O

When	NN	O	O
immunoglobulin	NN	O	B-cell_line
(	NN	O	I-cell_line
Ig	NN	O	I-cell_line
)	NN	O	I-cell_line
-secreting	NN	O	I-cell_line
plasmacytomas	NN	O	I-cell_line
are	NN	O	O
fused	NN	O	O
to	NN	O	O
a	NN	O	O
T-cell	NN	O	B-cell_type
lymphoma	NN	O	I-cell_type
,	NN	O	O
Ig	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
ceases	NN	O	O
in	NN	O	O
greater	NN	O	O
than	NN	O	O
95	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
resulting	NN	O	O
hybrids	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
rare	NN	O	O
hybrids	NN	O	O
that	NN	O	O
continue	NN	O	O
to	NN	O	O
express	NN	O	O
Ig	NN	O	O
,	NN	O	O
all	NN	O	O
other	NN	O	O
tested	NN	O	O
B	NN	O	B-DNA
lymphocyte-specific	NN	O	I-DNA
genes	NN	O	I-DNA
also	NN	O	O
remain	NN	O	O
active	NN	O	O
.	NN	O	O

The	NN	O	O
low	NN	O	O
frequency	NN	O	O
with	NN	O	O
which	NN	O	O
these	NN	O	O
Ig-expressing	NN	O	B-cell_line
hybrids	NN	O	I-cell_line
are	NN	O	O
recovered	NN	O	O
,	NN	O	O
along	NN	O	O
with	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
cell	NN	O	O
fusions	NN	O	O
can	NN	O	O
lead	NN	O	O
to	NN	O	O
chromosome	NN	O	O
loss	NN	O	O
,	NN	O	O
led	NN	O	O
us	NN	O	O
to	NN	O	O
propose	NN	O	O
that	NN	O	O
this	NN	O	O
rare	NN	O	O
phenotype	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
loss	NN	O	O
of	NN	O	O
a	NN	O	O
T-cell-derived	NN	O	B-protein
chromosome	NN	O	I-protein
encoding	NN	O	I-protein
a	NN	O	I-protein
factor	NN	O	I-protein
or	NN	O	O
factors	NN	O	O
with	NN	O	O
gene	NN	O	O
silencing	NN	O	O
activity	NN	O	O
.	NN	O	O

To	NN	O	O
identify	NN	O	O
the	NN	O	O
relevant	NN	O	O
chromosome	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
used	NN	O	O
a	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
-assisted	NN	O	O
method	NN	O	O
of	NN	O	O
chromosome	NN	O	O
mapping	NN	O	O
to	NN	O	O
analyze	NN	O	O
both	NN	O	O
Ig-silenced	NN	O	B-cell_line
(	NN	O	I-cell_line
common	NN	O	I-cell_line
)	NN	O	I-cell_line
and	NN	O	I-cell_line
Ig-expressing	NN	O	I-cell_line
(	NN	O	I-cell_line
rare	NN	O	I-cell_line
)	NN	O	I-cell_line
hybrids	NN	O	I-cell_line
.	NN	O	O

Although	NN	O	O
no	NN	O	O
single	NN	O	O
chromosome	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
correlate	NN	O	O
with	NN	O	O
Ig	NN	O	B-DNA
gene	NN	O	I-DNA
silencing	NN	O	O
,	NN	O	O
we	NN	O	O
discovered	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
types	NN	O	O
of	NN	O	O
hybrids	NN	O	O
had	NN	O	O
undergone	NN	O	O
distinct	NN	O	O
patterns	NN	O	O
of	NN	O	O
chromosome	NN	O	O
loss	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
Oct-2	NN	O	B-protein
)	NN	O	O
dramatically	NN	O	O
altered	NN	O	O
both	NN	O	O
the	NN	O	O
phenotype	NN	O	O
and	NN	O	O
chromosome	NN	O	O
constitution	NN	O	O
of	NN	O	O
hybrids	NN	O	O
arising	NN	O	O
in	NN	O	O
these	NN	O	O
cell	NN	O	O
fusions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reactivation	NN	O	O
of	NN	O	O
Kaposi	NN	O	O
's	NN	O	O
sarcoma-associated	NN	O	O
herpesvirus	NN	O	O
infection	NN	O	O
from	NN	O	O
latency	NN	O	O
by	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
ORF	NN	O	B-protein
50	NN	O	I-protein
transactivator	NN	O	I-protein
,	NN	O	O
a	NN	O	O
homolog	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
R	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Kaposi	NN	O	O
's	NN	O	O
sarcoma	NN	O	O
(	NN	O	O
KS	NN	O	O
)	NN	O	O
-associated	NN	O	O
herpesvirus	NN	O	O
(	NN	O	O
KSHV	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
8	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
lymphotropic	NN	O	O
virus	NN	O	O
strongly	NN	O	O
linked	NN	O	O
to	NN	O	O
several	NN	O	O
AIDS-related	NN	O	O
neoplasms	NN	O	O
.	NN	O	O

The	NN	O	O
primary	NN	O	O
reservoir	NN	O	O
of	NN	O	O
infection	NN	O	O
consists	NN	O	O
of	NN	O	O
latently	NN	O	O
infected	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
possibly	NN	O	O
other	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Viral	NN	O	O
reactivation	NN	O	O
from	NN	O	O
latency	NN	O	O
and	NN	O	O
spread	NN	O	O
from	NN	O	O
this	NN	O	O
lymphoid	NN	O	O
reservoir	NN	O	O
is	NN	O	O
presumably	NN	O	O
required	NN	O	O
for	NN	O	O
development	NN	O	O
of	NN	O	O
nonlymphoid	NN	O	O
tumors	NN	O	O
like	NN	O	O
KS	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
deregulated	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
viral	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
ORF	NN	O	B-DNA
50	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
a	NN	O	O
transactivator	NN	O	O
able	NN	O	O
to	NN	O	O
selectively	NN	O	O
upregulate	NN	O	O
delayed-early	NN	O	B-DNA
viral	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
suffices	NN	O	O
to	NN	O	O
disrupt	NN	O	O
latency	NN	O	O
and	NN	O	O
induce	NN	O	O
the	NN	O	O
lytic	NN	O	O
gene	NN	O	O
cascade	NN	O	O
in	NN	O	O
latently	NN	O	B-cell_type
infected	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
opens	NN	O	O
the	NN	O	O
way	NN	O	O
to	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
physiologic	NN	O	O
mechanisms	NN	O	O
controlling	NN	O	O
reactvation	NN	O	O
of	NN	O	O
KSHV	NN	O	O
from	NN	O	O
latency	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
are	NN	O	O
insufficient	NN	O	O
for	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
:	NN	O	O
requirement	NN	O	O
for	NN	O	O
CD28	NN	O	B-protein
up-regulation	NN	O	O
of	NN	O	O
RE/AP	NN	O	B-protein
.	NN	O	O

IL-2	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
both	NN	O	O
TCR	NN	O	O
and	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
.	NN	O	O

IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
stimulated	NN	O	O
with	NN	O	O
superantigen	NN	O	O
presented	NN	O	O
by	NN	O	O
Raji	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
requires	NN	O	O
CD28	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
rCTLA4Ig	NN	O	O
,	NN	O	O
which	NN	O	O
blocks	NN	O	O
CD28	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
its	NN	O	O
ligand	NN	O	O
,	NN	O	O
to	NN	O	O
the	NN	O	O
cultures	NN	O	O
decreased	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
by	NN	O	O
>	NN	O	O
80	NN	O	O
%	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
CTLA4Ig	NN	O	O
did	NN	O	O
not	NN	O	O
significantly	NN	O	O
inhibit	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
either	NN	O	O
NF	NN	O	B-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
or	NN	O	O
AP-1	NN	O	B-DNA
reporters	NN	O	I-DNA
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
insufficient	NN	O	O
for	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
an	NN	O	O
RE/AP	NN	O	B-protein
reporter	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
CTLA4Ig	NN	O	O
by	NN	O	O
>	NN	O	O
90	NN	O	O
%	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
requirement	NN	O	O
for	NN	O	O
CD28	NN	O	B-protein
in	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
RE/AP	NN	O	B-protein
and	NN	O	O
not	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
or	NN	O	I-DNA
AP-1	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
RE/AP	NN	O	B-protein
is	NN	O	O
not	NN	O	O
mediated	NN	O	O
by	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
because	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
NFAT	NN	O	B-protein
reporter	NN	O	O
is	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
CTLA4Ig	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
control	NN	O	O
of	NN	O	O
lytic	NN	O	O
replication	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
Review	NN	O	O
)	NN	O	O
.	NN	O	O

Uncontrolled	NN	O	O
replication	NN	O	O
of	NN	O	O
a	NN	O	O
virus	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
harmful	NN	O	O
to	NN	O	O
the	NN	O	O
host	NN	O	O
is	NN	O	O
also	NN	O	O
disadvantageous	NN	O	O
to	NN	O	O
the	NN	O	O
virus	NN	O	O
.	NN	O	O

Most	NN	O	O
viruses	NN	O	O
can	NN	O	O
not	NN	O	O
compete	NN	O	O
with	NN	O	O
the	NN	O	O
various	NN	O	O
immune	NN	O	O
mechanisms	NN	O	O
and	NN	O	O
become	NN	O	O
eliminated	NN	O	O
in	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
infection	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
only	NN	O	O
the	NN	O	O
time	NN	O	O
between	NN	O	O
infection	NN	O	O
and	NN	O	O
eradication	NN	O	O
remains	NN	O	O
for	NN	O	O
these	NN	O	O
viruses	NN	O	O
to	NN	O	O
proliferate	NN	O	O
.	NN	O	O

A	NN	O	O
few	NN	O	O
viruses	NN	O	O
,	NN	O	O
like	NN	O	O
the	NN	O	O
Herpesviruses	NN	O	O
or	NN	O	O
the	NN	O	O
papillomaviruses	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
sophisticated	NN	O	O
strategy	NN	O	O
for	NN	O	O
persisting	NN	O	O
lifelong	NN	O	O
,	NN	O	O
usually	NN	O	O
asymptomatically	NN	O	O
in	NN	O	O
the	NN	O	O
host	NN	O	O
,	NN	O	O
hiding	NN	O	O
from	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
and	NN	O	O
producing	NN	O	O
infectious	NN	O	O
progeny	NN	O	O
at	NN	O	O
the	NN	O	O
same	NN	O	O
time	NN	O	O
.	NN	O	O

This	NN	O	O
strategy	NN	O	O
depends	NN	O	O
on	NN	O	O
a	NN	O	O
separation	NN	O	O
of	NN	O	O
latency	NN	O	O
and	NN	O	O
the	NN	O	O
lytic	NN	O	O
replication	NN	O	O
,	NN	O	O
either	NN	O	O
by	NN	O	O
time	NN	O	O
due	NN	O	O
to	NN	O	O
differentiation-dependent	NN	O	O
mechanisms	NN	O	O
or	NN	O	O
by	NN	O	O
spatial	NN	O	O
separation	NN	O	O
as	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
different	NN	O	O
host	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Both	NN	O	O
are	NN	O	O
true	NN	O	O
for	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
.	NN	O	O

B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
have	NN	O	O
a	NN	O	O
pivotal	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
life	NN	O	O
cycle	NN	O	O
of	NN	O	O
the	NN	O	O
virus	NN	O	O
.	NN	O	O

The	NN	O	O
former	NN	O	O
can	NN	O	O
become	NN	O	O
latently	NN	O	O
infected	NN	O	O
and	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
virus	NN	O	O
reservoir	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
latter	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
permissive	NN	O	O
for	NN	O	O
lytic	NN	O	O
replication	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
replication	NN	O	O
of	NN	O	O
EBV	NN	O	O
in	NN	O	O
vivo	NN	O	O
is	NN	O	O
controlled	NN	O	O
primarily	NN	O	O
by	NN	O	O
host	NN	O	O
immune	NN	O	O
mechanisms	NN	O	O
selecting	NN	O	O
for	NN	O	O
cells	NN	O	O
that	NN	O	O
are	NN	O	O
not	NN	O	O
permissive	NN	O	O
for	NN	O	O
viral	NN	O	O
replication	NN	O	O
as	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
particular	NN	O	O
set	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

These	NN	O	O
factors	NN	O	O
control	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
regulatory	NN	O	B-DNA
immediate-early	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
,	NN	O	O
lytic	NN	O	O
and	NN	O	O
latent	NN	O	O
cycle	NN	O	O
regulatory	NN	O	O
genes	NN	O	O
negatively	NN	O	O
interfere	NN	O	O
with	NN	O	O
each	NN	O	O
other	NN	O	O
and	NN	O	O
thus	NN	O	O
link	NN	O	O
cellular	NN	O	O
and	NN	O	O
viral	NN	O	B-DNA
gene	NN	O	I-DNA
regulatory	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

Disturbance	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
immune	NN	O	O
surveillance	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
viral	NN	O	B-DNA
gene	NN	O	I-DNA
regulation	NN	O	O
may	NN	O	O
result	NN	O	O
in	NN	O	O
EBV-associated	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Adaptor	NN	O	O
function	NN	O	O
for	NN	O	O
the	NN	O	O
Syk	NN	O	B-protein
kinases-interacting	NN	O	I-protein
protein	NN	O	I-protein
3BP2	NN	O	B-protein
in	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
activation	NN	O	O
.	NN	O	O

Syk-family	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
are	NN	O	O
essential	NN	O	O
for	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
and	NN	O	O
activation	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
yeast	NN	O	O
two-hybrid	NN	O	O
screen	NN	O	O
to	NN	O	O
identify	NN	O	O
Syk	NN	O	B-protein
kinases-interacting	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
SKIPs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
3BP2	NN	O	B-protein
,	NN	O	O
an	NN	O	O
Abl	NN	O	B-protein
SH3-interacting	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
unknown	NN	O	O
function	NN	O	O
.	NN	O	O

3BP2	NN	O	B-protein
was	NN	O	O
selectively	NN	O	O
expressed	NN	O	O
in	NN	O	O
hematopoietic/lymphoid	NN	O	O
tissues	NN	O	O
and	NN	O	O
bound	NN	O	O
via	NN	O	O
its	NN	O	O
SH2	NN	O	B-protein
domain	NN	O	I-protein
activated	NN	O	I-protein
Syk-family	NN	O	I-protein
kinases	NN	O	I-protein
in	NN	O	O
mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
including	NN	O	O
in	NN	O	O
antigen	NN	O	B-cell_type
receptor-stimulated	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
Zap-70	NN	O	B-protein
,	NN	O	O
the	NN	O	O
3BP2	NN	O	B-protein
SH2	NN	O	I-protein
domain	NN	O	I-protein
associated	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
LAT	NN	O	B-protein
,	NN	O	O
Grb2	NN	O	B-protein
,	NN	O	O
PLCgamma1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Cbl	NN	O	B-protein
from	NN	O	O
activated	NN	O	O
T	NN	O	O
cell	NN	O	O
lysates	NN	O	O
.	NN	O	O

Transient	NN	O	O
3BP2	NN	O	B-protein
overexpression	NN	O	O
induced	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
its	NN	O	O
NFAT	NN	O	B-DNA
or	NN	O	I-DNA
AP-1	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
activity	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
SH2	NN	O	B-protein
and	NN	O	O
pleckstrin-homology	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
3BP2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
required	NN	O	O
functional	NN	O	O
Syk	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
Ras	NN	O	B-protein
,	NN	O	O
and	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
3BP2	NN	O	B-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
adaptor	NN	O	O
that	NN	O	O
may	NN	O	O
couple	NN	O	O
activated	NN	O	O
Zap-70/Syk	NN	O	B-protein
to	NN	O	O
a	NN	O	O
LAT-containing	NN	O	B-protein
signaling	NN	O	I-protein
complex	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
TCR-mediated	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vitro	NN	O	O
suppression	NN	O	O
of	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
tissue	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
metalloproteinases-1	NN	O	B-protein
.	NN	O	O

Cellular	NN	O	O
pathways	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
(	NN	O	O
PCD	NN	O	O
)	NN	O	O
have	NN	O	O
been	NN	O	O
identified	NN	O	O
,	NN	O	O
but	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
specific	NN	O	O
extracellular	NN	O	O
matrix	NN	O	O
processes	NN	O	O
that	NN	O	O
may	NN	O	O
affect	NN	O	O
apoptosis	NN	O	O
along	NN	O	O
those	NN	O	O
pathways	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
Burkitt	NN	O	B-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
(	NN	O	I-cell_line
BL	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
assayed	NN	O	O
for	NN	O	O
their	NN	O	O
expression	NN	O	O
of	NN	O	O
tissue	NN	O	B-protein
inhibitor	NN	O	I-protein
of	NN	O	I-protein
metalloproteinases	NN	O	I-protein
(	NN	O	I-protein
TIMP	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
.	NN	O	O

Results	NN	O	O
indicate	NN	O	O
that	NN	O	O
TIMP-1-positive	NN	O	B-cell_line
BL	NN	O	I-cell_line
lines	NN	O	I-cell_line
show	NN	O	O
resistance	NN	O	O
to	NN	O	O
cold-shock-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
recombinant	NN	O	O
TIMP-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
TIMP-2	NN	O	B-protein
or	NN	O	O
a	NN	O	O
synthetic	NN	O	O
metalloproteinase	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
BB-94	NN	O	O
)	NN	O	O
,	NN	O	O
confers	NN	O	O
resistance	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
both	NN	O	O
CD95-dependent	NN	O	O
and	NN	O	O
-independent	NN	O	O
(	NN	O	O
cold	NN	O	O
shock	NN	O	O
,	NN	O	O
serum	NN	O	O
deprivation	NN	O	O
,	NN	O	O
and	NN	O	O
gamma-radiation	NN	O	O
)	NN	O	O
pathways	NN	O	O
in	NN	O	O
TIMP-1-negative	NN	O	B-cell_line
BL	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

TIMP-1	NN	O	B-protein
suppression	NN	O	O
of	NN	O	O
PCD	NN	O	O
is	NN	O	O
not	NN	O	O
due	NN	O	O
to	NN	O	O
metalloproteinase	NN	O	B-protein
inhibition	NN	O	O
,	NN	O	O
as	NN	O	O
reduction	NN	O	O
and	NN	O	O
alkylation	NN	O	O
of	NN	O	O
the	NN	O	O
TIMP-1	NN	O	B-protein
did	NN	O	O
not	NN	O	O
abolish	NN	O	O
this	NN	O	O
activity	NN	O	O
.	NN	O	O

Retroviral	NN	O	O
induction	NN	O	O
of	NN	O	O
TIMP-1	NN	O	B-protein
not	NN	O	O
only	NN	O	O
resulted	NN	O	O
in	NN	O	O
cell	NN	O	O
survival	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
continued	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
5	NN	O	O
d	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
serum	NN	O	O
,	NN	O	O
while	NN	O	O
controls	NN	O	O
underwent	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

This	NN	O	O
resistance	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
reversed	NN	O	O
by	NN	O	O
anti-TIMP-1	NN	O	B-protein
antibodies	NN	O	I-protein
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
secreted	NN	O	O
TIMP-1	NN	O	B-protein
is	NN	O	O
active	NN	O	O
in	NN	O	O
blocking	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
TIMP-1	NN	O	B-protein
upregulation	NN	O	O
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
Bcl-XL	NN	O	B-protein
but	NN	O	O
not	NN	O	O
Bcl-2	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
decreased	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
controls	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
TIMP-1	NN	O	B-protein
suppresses	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
suggests	NN	O	O
a	NN	O	O
novel	NN	O	O
activity	NN	O	O
for	NN	O	O
TIMP-1	NN	O	B-protein
in	NN	O	O
tissue	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Spi-1/PU.1	NN	O	B-DNA
proto-oncogene	NN	O	B-DNA
induces	NN	O	O
opposite	NN	O	O
effects	NN	O	O
on	NN	O	O
monocytic	NN	O	O
and	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Spi-1/PU.1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
hematopoietic	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

To	NN	O	O
analyze	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
spi-1	NN	O	B-protein
on	NN	O	O
the	NN	O	O
proliferation/differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
spi-1	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
transfected	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
expressed	NN	O	O
elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
spi-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
and	NN	O	O
high	NN	O	O
Spi-1	NN	O	B-protein
-DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
spi-1	NN	O	B-cell_line
transfected	NN	O	I-cell_line
cells	NN	O	I-cell_line
showed	NN	O	O
reduced	NN	O	O
growth	NN	O	O
rates	NN	O	O
and	NN	O	O
reduced	NN	O	O
clonogenic	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

When	NN	O	O
the	NN	O	O
erythroid	NN	O	O
and	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
markers	NN	O	O
were	NN	O	O
analyzed	NN	O	O
,	NN	O	O
spi-1	NN	O	B-protein
overexpression	NN	O	O
resulted	NN	O	O
in	NN	O	O
opposite	NN	O	O
effects	NN	O	O
:	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
was	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
in	NN	O	O
spi-1	NN	O	B-cell_line
transfectants	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
spi-1	NN	O	B-protein
overexpression	NN	O	O
increased	NN	O	O
the	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
a	NN	O	O
differential	NN	O	O
role	NN	O	O
of	NN	O	O
Spi-1	NN	O	B-DNA
on	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
function	NN	O	O
of	NN	O	O
an	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
receptor	NN	O	I-protein
beta	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	B-protein
IL-2Rbeta	NN	O	I-protein
)	NN	O	I-protein
/IL-4Ralpha	NN	O	I-protein
cytokine	NN	O	I-protein
receptor	NN	O	I-protein
chimera	NN	O	I-protein
potentiates	NN	O	O
allergic	NN	O	O
airway	NN	O	O
disease	NN	O	O
.	NN	O	O

Strength	NN	O	O
of	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
,	NN	O	O
coreceptors	NN	O	B-protein
,	NN	O	O
costimulation	NN	O	O
,	NN	O	O
antigen-presenting	NN	O	B-cell_type
cell	NN	O	I-cell_type
type	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
cytokines	NN	O	O
all	NN	O	O
play	NN	O	O
crucial	NN	O	O
roles	NN	O	O
in	NN	O	O
determining	NN	O	O
the	NN	O	O
efficiency	NN	O	O
with	NN	O	O
which	NN	O	O
type	NN	O	B-cell_type
2	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
Th2	NN	O	B-cell_type
,	NN	O	O
Tc2	NN	O	B-cell_type
)	NN	O	O
develop	NN	O	O
from	NN	O	O
uncommitted	NN	O	B-protein
precursors	NN	O	I-protein
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
in	NN	O	O
vivo	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
the	NN	O	O
population	NN	O	O
of	NN	O	O
type	NN	O	B-cell_type
2	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
disease	NN	O	O
susceptibility	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
created	NN	O	O
lines	NN	O	O
of	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
in	NN	O	O
which	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
chimeric	NN	O	B-protein
cytokine	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
the	NN	O	O
mouse	NN	O	B-protein
interleukin	NN	O	I-protein
2	NN	O	I-protein
receptor	NN	O	I-protein
beta	NN	O	I-protein
chain	NN	O	I-protein
[	NN	O	I-protein
IL-2Rbeta	NN	O	I-protein
]	NN	O	I-protein
extracellular	NN	O	I-protein
domain	NN	O	I-protein
fused	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
tail	NN	O	I-protein
of	NN	O	O
IL-4Ralpha	NN	O	B-protein
)	NN	O	O
is	NN	O	O
targeted	NN	O	O
to	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
lineage	NN	O	I-cell_type
using	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
lck	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
chimera	NN	O	O
transduced	NN	O	O
IL-4	NN	O	B-protein
-specific	NN	O	O
signals	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
binding	NN	O	O
and	NN	O	O
dramatically	NN	O	O
enhanced	NN	O	O
type	NN	O	O
2	NN	O	O
responses	NN	O	O
(	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-5	NN	O	B-protein
,	NN	O	O
and	NN	O	O
immunoglobulin	NN	O	B-protein
E	NN	O	I-protein
production	NN	O	O
)	NN	O	O
upon	NN	O	O
in	NN	O	O
vitro	NN	O	O
TCR	NN	O	B-protein
stimulation	NN	O	O
or	NN	O	O
in	NN	O	O
vivo	NN	O	O
antigen	NN	O	O
challenge	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
type	NN	O	O
2	NN	O	O
effector	NN	O	O
function	NN	O	O
was	NN	O	O
augmented	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
signals	NN	O	O
transduced	NN	O	O
through	NN	O	O
a	NN	O	O
chimeric	NN	O	B-protein
receptor	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
T	NN	O	O
cell-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

This	NN	O	O
influence	NN	O	O
was	NN	O	O
sufficient	NN	O	O
for	NN	O	O
establishment	NN	O	O
of	NN	O	O
antigen-induced	NN	O	O
allergic	NN	O	O
airway	NN	O	O
hyperresponsiveness	NN	O	O
on	NN	O	O
a	NN	O	O
disease-resistant	NN	O	O
background	NN	O	O

-DOCSTART-	O

Differentiation-dependent	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-protein
carboxylesterase	NN	O	I-protein
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

Carboxylesterases	NN	O	B-protein
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
defense	NN	O	O
and	NN	O	O
clearance	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	O
system	NN	O	O
.	NN	O	O

During	NN	O	O
the	NN	O	O
differentiation	NN	O	O
process	NN	O	O
of	NN	O	O
cells	NN	O	O
from	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
we	NN	O	O
observed	NN	O	O
a	NN	O	O
transient	NN	O	O
transcriptional	NN	O	O
upregulation	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-protein
carboxylesterase	NN	O	I-protein
analyzed	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
Northern	NN	O	O
blots	NN	O	O
.	NN	O	O

In	NN	O	O
PMA-treated	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
we	NN	O	O
could	NN	O	O
detect	NN	O	O
three	NN	O	O
major	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
sites	NN	O	I-DNA
as	NN	O	O
revealed	NN	O	O
by	NN	O	O
Nuclease	NN	O	O
Protection	NN	O	O
Assay	NN	O	O
carried	NN	O	O
out	NN	O	O
with	NN	O	O
two	NN	O	O
overlapping	NN	O	B-RNA
antisense	NN	O	I-RNA
RNA	NN	O	I-RNA
probes	NN	O	I-RNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
cloned	NN	O	O
the	NN	O	O
carboxylesterase	NN	O	B-DNA
upstream	NN	O	I-DNA
sequence	NN	O	I-DNA
and	NN	O	O
showed	NN	O	O
its	NN	O	O
basal	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
CHO	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analysis	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
spanning	NN	O	O
base	NN	O	B-DNA
pairs	NN	O	I-DNA
-1	NN	O	I-DNA
to	NN	O	I-DNA
-275	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
contains	NN	O	O
several	NN	O	O
putative	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
is	NN	O	O
bound	NN	O	O
by	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
Sp1	NN	O	B-protein
and	NN	O	O
IRBP	NN	O	B-protein
but	NN	O	O
not	NN	O	O
by	NN	O	O
C/EBPs	NN	O	B-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
these	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
carboxylesterase	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
starts	NN	O	O
at	NN	O	O
multiple	NN	O	O
initiation	NN	O	O
sites	NN	O	O
and	NN	O	O
that	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
IRBP	NN	O	B-protein
may	NN	O	O
be	NN	O	O
critical	NN	O	O
factors	NN	O	O
for	NN	O	O
modulating	NN	O	O
the	NN	O	O
differentiation-dependent	NN	O	O
transcription	NN	O	O
of	NN	O	O
this	NN	O	O
human	NN	O	B-DNA
carboxylesterase	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
Ikaros	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
and	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

The	NN	O	O
Ikaros	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
hemopoietic-specific	NN	O	B-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
is	NN	O	O
described	NN	O	O
as	NN	O	O
a	NN	O	O
central	NN	O	O
regulator	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

During	NN	O	O
fetal	NN	O	O
development	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
required	NN	O	O
at	NN	O	O
the	NN	O	O
earliest	NN	O	O
stage	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
and	NN	O	O
B	NN	O	O
cell	NN	O	O
specification	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
adult	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
lymphoid	NN	O	B-cell_type
lineages	NN	O	I-cell_type
rely	NN	O	O
on	NN	O	O
Ikaros	NN	O	B-DNA
at	NN	O	O
distinct	NN	O	O
phases	NN	O	O
of	NN	O	O
their	NN	O	O
development	NN	O	O
.	NN	O	O

Its	NN	O	O
activity	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
precursors	NN	O	I-cell_type
,	NN	O	O
although	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
latter	NN	O	O
is	NN	O	O
not	NN	O	O
normal	NN	O	O
.	NN	O	O

A	NN	O	O
significant	NN	O	O
increase	NN	O	O
in	NN	O	O
CD4	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
and	NN	O	O
their	NN	O	O
immediate	NN	O	O
precursors	NN	O	O
is	NN	O	O
detected	NN	O	O
,	NN	O	O
and	NN	O	O
because	NN	O	O
these	NN	O	O
cells	NN	O	O
lack	NN	O	O
markers	NN	O	O
that	NN	O	O
correlate	NN	O	O
with	NN	O	O
positive	NN	O	O
selection	NN	O	O
,	NN	O	O
a	NN	O	O
deregulation	NN	O	O
in	NN	O	O
their	NN	O	O
maturation	NN	O	O
process	NN	O	O
is	NN	O	O
suggested	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
Ikaros-null	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
hyperproliferate	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
;	NN	O	O
within	NN	O	O
days	NN	O	O
after	NN	O	O
their	NN	O	O
appearance	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
,	NN	O	O
clonally	NN	O	O
expanding	NN	O	O
populations	NN	O	O
are	NN	O	O
detected	NN	O	O
.	NN	O	O

Deregulated	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
responses	NN	O	O
and	NN	O	O
the	NN	O	O
fast	NN	O	O
kinetics	NN	O	O
of	NN	O	O
tumor	NN	O	O
development	NN	O	O
in	NN	O	O
these	NN	O	O
mutant	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
implicate	NN	O	O
Ikaros	NN	O	O
as	NN	O	O
a	NN	O	O
central	NN	O	B-DNA
tumor	NN	O	I-DNA
suppressor	NN	O	I-DNA
gene	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
lack	NN	O	O
of	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
selective	NN	O	O
defects	NN	O	O
in	NN	O	O
gamma	NN	O	B-cell_type
delta	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
dendritic	NN	O	B-cell_type
antigen-presenting	NN	O	I-cell_type
cells	NN	O	I-cell_type
point	NN	O	O
to	NN	O	O
Ikaros	NN	O	B-protein
as	NN	O	O
an	NN	O	O
essential	NN	O	O
factor	NN	O	O
for	NN	O	O
the	NN	O	O
establishment	NN	O	O
of	NN	O	O
early	NN	O	O
branchpoints	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
pathway	NN	O	O
.	NN	O	O

The	NN	O	O
dominant	NN	O	O
interference	NN	O	O
activity	NN	O	O
of	NN	O	O
Ikaros	NN	O	B-protein
isoforms	NN	O	I-protein
unable	NN	O	O
to	NN	O	O
bind	NN	O	O
DNA	NN	O	O
and	NN	O	O
their	NN	O	O
effects	NN	O	O
in	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
suggest	NN	O	O
that	NN	O	O
Ikaros	NN	O	B-protein
works	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
other	NN	O	O
factors	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
Aiolos	NN	O	B-DNA
,	NN	O	O
a	NN	O	O
lymphoid-restricted	NN	O	B-DNA
and	NN	O	I-DNA
structurally	NN	O	I-DNA
related	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
differentiation	NN	O	O
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

A	NN	O	O
model	NN	O	O
is	NN	O	O
proposed	NN	O	O
that	NN	O	O
defines	NN	O	O
Ikaros	NN	O	B-protein
as	NN	O	O
the	NN	O	O
backbone	NN	O	O
of	NN	O	O
a	NN	O	O
complex	NN	O	B-protein
regulatory	NN	O	I-protein
protein	NN	O	I-protein
network	NN	O	I-protein
that	NN	O	O
controls	NN	O	O
cell	NN	O	O
fate	NN	O	O
decisions	NN	O	O
and	NN	O	O
regulates	NN	O	O
homeostasis	NN	O	O
in	NN	O	O
the	NN	O	O
hemo-lymphoid	NN	O	O
system	NN	O	O
.	NN	O	O

Changes	NN	O	O
in	NN	O	O
this	NN	O	O
regulatory	NN	O	O
network	NN	O	O
may	NN	O	O
reflect	NN	O	O
differentiation	NN	O	O
and	NN	O	O
proliferation	NN	O	O
adjustments	NN	O	O
made	NN	O	O
in	NN	O	O
hemo-lymphoid	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
and	NN	O	O
precursors	NN	O	B-cell_type
as	NN	O	O
they	NN	O	O
give	NN	O	O
rise	NN	O	O
to	NN	O	O
the	NN	O	O
cells	NN	O	O
of	NN	O	O
our	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

-DOCSTART-	O

STAT1	NN	O	B-protein
pathway	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
caprine	NN	O	O
arthritis-encephalitis	NN	O	B-DNA
virus	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
caprine	NN	O	B-DNA
arthritis-encephalitis	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
CAEV	NN	O	I-DNA
)	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
is	NN	O	O
activated	NN	O	O
by	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
in	NN	O	O
promonocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
a	NN	O	O
70-bp	NN	O	B-DNA
element	NN	O	I-DNA
is	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
CAEV	NN	O	B-DNA
LTR	NN	O	I-DNA
to	NN	O	O
this	NN	O	O
cytokine	NN	O	B-protein
.	NN	O	O

At	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
end	NN	O	I-DNA
,	NN	O	O
this	NN	O	O
70-bp	NN	O	B-DNA
IFN-gamma	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
contains	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
to	NN	O	O
the	NN	O	O
gamma	NN	O	B-DNA
activated	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
GAS	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
putative	NN	O	O
GAS	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
CAEV	NN	O	B-DNA
LTR	NN	O	I-DNA
binds	NN	O	O
specifically	NN	O	O
to	NN	O	O
a	NN	O	O
cellular	NN	O	B-protein
factor	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
promonocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Substitution	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
consensus	NN	O	B-DNA
sequence	NN	O	I-DNA
eliminate	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
GAS	NN	O	B-DNA
element	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
70-bp	NN	O	B-DNA
motif	NN	O	I-DNA
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
confer	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
using	NN	O	O
a	NN	O	O
heterologous	NN	O	O
minimal	NN	O	O
promoter	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
binding	NN	O	O
data	NN	O	O
,	NN	O	O
the	NN	O	O
same	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
GAS	NN	O	B-DNA
element	NN	O	I-DNA
eliminate	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
both	NN	O	O
a	NN	O	O
functional	NN	O	O
CAEV	NN	O	B-DNA
LTR	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
cellular	NN	O	O
factor	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
GAS	NN	O	B-DNA
element	NN	O	I-DNA
is	NN	O	O
present	NN	O	O
from	NN	O	O
5	NN	O	O
min	NN	O	O
to	NN	O	O
14	NN	O	O
h	NN	O	O
after	NN	O	O
stimulation	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
to	NN	O	O
the	NN	O	O
GAS	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
CAEV	NN	O	B-DNA
LTR	NN	O	I-DNA
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
antibody	NN	O	O
directed	NN	O	O
against	NN	O	O
STAT1	NN	O	B-protein
(	NN	O	O
p91/84	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
GAS	NN	O	O
sequence	NN	O	O
in	NN	O	O
the	NN	O	O
CAEV	NN	O	B-DNA
LTR	NN	O	I-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
a	NN	O	O
STAT1-like	NN	O	B-protein
factor	NN	O	I-protein
binds	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
.	NN	O	O

The	NN	O	O
STAT-1	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
provides	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
mechanism	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
CAEV	NN	O	B-DNA
LTR	NN	O	I-DNA
by	NN	O	O
IFN-gamma	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
the	NN	O	O
first	NN	O	O
demonstration	NN	O	O
of	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
a	NN	O	O
STAT	NN	O	B-protein
family	NN	O	I-protein
member	NN	O	I-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
a	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

1	NN	O	B-DNA
,	NN	O	I-DNA
25-Dihydroxyvitamin	NN	O	I-DNA
D3	NN	O	I-DNA
receptors	NN	O	I-DNA
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
renal	NN	O	O
insufficiency	NN	O	O
.	NN	O	O

A	NN	O	O
reduced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
VDR	NN	O	O
)	NN	O	O
in	NN	O	O
parathyroid	NN	O	O
glands	NN	O	O
of	NN	O	O
uremic	NN	O	O
animals	NN	O	O
and	NN	O	O
humans	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
.	NN	O	O

Similar	NN	O	O
results	NN	O	O
have	NN	O	O
been	NN	O	O
obtained	NN	O	O
by	NN	O	O
our	NN	O	O
own	NN	O	O
group	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
secondary	NN	O	O
hyperparathyroidism	NN	O	O
to	NN	O	O
chronic	NN	O	O
renal	NN	O	O
failure	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
reasons	NN	O	O
for	NN	O	O
these	NN	O	O
changes	NN	O	O
are	NN	O	O
not	NN	O	O
clear	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
specific	NN	O	O
uptake	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
by	NN	O	O
PBMC	NN	O	B-cell_type
of	NN	O	O
11	NN	O	O
women	NN	O	O
with	NN	O	O
advanced	NN	O	O
chronic	NN	O	O
renal	NN	O	O
failure	NN	O	O
(	NN	O	O
A-CRF	NN	O	O
)	NN	O	O
,	NN	O	O
6	NN	O	O
women	NN	O	O
with	NN	O	O
mild-moderate	NN	O	O
renal	NN	O	O
insufficiency	NN	O	O
(	NN	O	O
M-CRF	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
23	NN	O	O
healthy	NN	O	O
women	NN	O	O
.	NN	O	O

The	NN	O	O
mean	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
KD	NN	O	O
)	NN	O	O
was	NN	O	O
similar	NN	O	O
in	NN	O	O
both	NN	O	O
groups	NN	O	O
of	NN	O	O
patients	NN	O	O
and	NN	O	O
in	NN	O	O
healthy	NN	O	O
women	NN	O	O
(	NN	O	O
A-CRF	NN	O	O
:	NN	O	O
0.7	NN	O	O
+/-	NN	O	O
0.5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
M	NN	O	O
;	NN	O	O
M-CRF	NN	O	O
:	NN	O	O
1.1	NN	O	O
+/-	NN	O	O
0.9	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
M	NN	O	O
;	NN	O	O
controls	NN	O	O
:	NN	O	O
1.0	NN	O	O
+/-	NN	O	O
0.6	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
M	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
VDR	NN	O	B-protein
concentration	NN	O	O
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
in	NN	O	O
A-CRF	NN	O	O
(	NN	O	O
0.8	NN	O	O
+/-	NN	O	O
0.5	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
vs.	NN	O	O
2.3	NN	O	O
+/-	NN	O	O
0.9	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
controls	NN	O	O
,	NN	O	O
p	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
no	NN	O	O
changes	NN	O	O
were	NN	O	O
seen	NN	O	O
in	NN	O	O
M-CRF	NN	O	O
(	NN	O	O
1.7	NN	O	O
+/-	NN	O	O
0.7	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
vs.	NN	O	O
2.3	NN	O	O
+/-	NN	O	O
0.9	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
cells	NN	O	O
in	NN	O	O
controls	NN	O	O
)	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
was	NN	O	O
seen	NN	O	O
between	NN	O	O
VDR	NN	O	B-protein
and	NN	O	O
serum	NN	O	O
calcitriol	NN	O	O
or	NN	O	O
PTH	NN	O	O
levels	NN	O	O
,	NN	O	O
when	NN	O	O
considering	NN	O	O
both	NN	O	O
groups	NN	O	O
of	NN	O	O
patients	NN	O	O
together	NN	O	O
or	NN	O	O
separately	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
negative	NN	O	O
correlation	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
VDR	NN	O	B-protein
and	NN	O	O
serum	NN	O	O
creatinine	NN	O	O
values	NN	O	O
when	NN	O	O
A-CRF	NN	O	O
and	NN	O	O
M-CRF	NN	O	O
were	NN	O	O
considered	NN	O	O
altogether	NN	O	O
(	NN	O	O
r	NN	O	O
=	NN	O	O
-0.63	NN	O	O
;	NN	O	O
p	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
two	NN	O	O
different	NN	O	O
schedules	NN	O	O
of	NN	O	O
oral	NN	O	O
calcitriol	NN	O	O
(	NN	O	O
five	NN	O	O
patients	NN	O	O
with	NN	O	O
0.5	NN	O	O
microgram/day	NN	O	O
for	NN	O	O
1	NN	O	O
month	NN	O	O
and	NN	O	O
four	NN	O	O
patients	NN	O	O
with	NN	O	O
2	NN	O	O
micrograms/day	NN	O	O
for	NN	O	O
7	NN	O	O
days	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
change	NN	O	O
VDR	NN	O	B-protein
concentrations	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
serum	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
of	NN	O	O
uremia	NN	O	O
are	NN	O	O
not	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
decrease	NN	O	O
in	NN	O	O
VDR	NN	O	B-protein
concentration	NN	O	O
found	NN	O	O
in	NN	O	O
these	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
promoter	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
monoamine	NN	O	I-DNA
oxidase	NN	O	I-DNA
B	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
cerebral	NN	O	O
cortex	NN	O	O
and	NN	O	O
in	NN	O	O
blood	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Many	NN	O	O
studies	NN	O	O
show	NN	O	O
that	NN	O	O
monoamine	NN	O	B-protein
oxidase	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
blood	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
a	NN	O	O
biological	NN	O	O
marker	NN	O	O
for	NN	O	O
personality	NN	O	O
characteristics	NN	O	O
such	NN	O	O
as	NN	O	O
sensation	NN	O	O
seeking	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
underlying	NN	O	O
this	NN	O	O
association	NN	O	O
is	NN	O	O
so	NN	O	O
far	NN	O	O
not	NN	O	O
explored	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
have	NN	O	O
performed	NN	O	O
electrophoretic	NN	O	O
mobility-shift	NN	O	O
assays	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
protein	NN	O	O
binding	NN	O	O
to	NN	O	O
a	NN	O	O
150	NN	O	B-DNA
bp	NN	O	I-DNA
fragment	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
5'-flanking	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
monoamine	NN	O	I-DNA
oxidase	NN	O	I-DNA
B	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
compared	NN	O	O
the	NN	O	O
pattern	NN	O	O
using	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
human	NN	O	O
brain	NN	O	O
and	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
a	NN	O	O
correlation	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
monoamine	NN	O	B-protein
oxidase	NN	O	I-protein
B	NN	O	I-protein
enzyme	NN	O	O
activity	NN	O	O
in	NN	O	O
blood	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
platelets	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
the	NN	O	O
binding	NN	O	O
pattern	NN	O	O
of	NN	O	O
two	NN	O	O
uncharacterized	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
well	NN	O	O
in	NN	O	O
line	NN	O	O
with	NN	O	O
the	NN	O	O
long-standing	NN	O	O
notion	NN	O	O
that	NN	O	O
interindividual	NN	O	O
differences	NN	O	O
in	NN	O	O
platelet	NN	O	B-protein
monoamine	NN	O	I-protein
oxidase	NN	O	I-protein
may	NN	O	O
represent	NN	O	O
differences	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
rather	NN	O	O
than	NN	O	O
genotypic	NN	O	O
variation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Enhanced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
macrophages	NN	O	B-cell_type
induced	NN	O	O
by	NN	O	O
ciprofibrate	NN	O	O
.	NN	O	O

Ciprofibrate	NN	O	O
,	NN	O	O
an	NN	O	O
hypolipidaemic	NN	O	O
peroxisome	NN	O	O
proliferator	NN	O	O
,	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
effect	NN	O	O
was	NN	O	O
greatly	NN	O	O
potentiated	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
at	NN	O	O
a	NN	O	O
concentration	NN	O	O
where	NN	O	O
neither	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
nor	NN	O	O
ciprofibrate	NN	O	O
alone	NN	O	O
had	NN	O	O
any	NN	O	O
effect	NN	O	O
on	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

As	NN	O	O
occurs	NN	O	O
for	NN	O	O
HL-60	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
induced	NN	O	O
by	NN	O	O
high	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
concentration	NN	O	O
,	NN	O	O
the	NN	O	O
ciprofibrate-induced	NN	O	O
phorbol	NN	O	O
ester-dependent	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
proceeded	NN	O	O
through	NN	O	O
the	NN	O	O
monocytic/macrophage	NN	O	O
pathway	NN	O	O
and	NN	O	O
induced	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
proteins	NN	O	O
with	NN	O	O
similar	NN	O	O
molecular	NN	O	O
weights	NN	O	O
suggesting	NN	O	O
that	NN	O	O
increased	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
activity	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
effect	NN	O	O
.	NN	O	O

The	NN	O	O
peroxisome	NN	O	O
proliferator-activated	NN	O	O
receptor	NN	O	O
(	NN	O	O
PPARalpha	NN	O	O
)	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
no	NN	O	O
changes	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
its	NN	O	O
expression	NN	O	O
upon	NN	O	O
HL-60	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
multiple	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
TNFR	NN	O	I-protein
)	NN	O	I-protein
-associated	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
receptor	NN	O	B-protein
activator	NN	O	I-protein
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
TNFR	NN	O	B-protein
superfamily	NN	O	I-protein
.	NN	O	O

Receptor	NN	O	B-protein
activator	NN	O	I-protein
of	NN	O	I-protein
NF-kappaB	NN	O	I-protein
(	NN	O	O
RANK	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
recently	NN	O	O
identified	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
on	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Its	NN	O	O
cognate	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
RANKL	NN	O	B-protein
)	NN	O	O
plays	NN	O	O
significant	NN	O	O
roles	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
dendritic	NN	O	O
cell	NN	O	O
function	NN	O	O
and	NN	O	O
osteoclast	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
RANK	NN	O	B-protein
with	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-associated	NN	O	I-protein
factors	NN	O	I-protein
(	NN	O	I-protein
TRAFs	NN	O	I-protein
)	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	I-protein
2	NN	O	I-protein
,	NN	O	I-protein
3	NN	O	I-protein
,	NN	O	I-protein
5	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
6	NN	O	I-protein
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
cells	NN	O	O
.	NN	O	O

Mapping	NN	O	O
of	NN	O	O
the	NN	O	O
structural	NN	O	O
requirements	NN	O	O
for	NN	O	O
TRAF	NN	O	B-protein
/RANK	NN	O	B-protein
interaction	NN	O	O
revealed	NN	O	O
multiple	NN	O	O
TRAF	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
clustered	NN	O	O
in	NN	O	O
two	NN	O	O
distinct	NN	O	O
domains	NN	O	O
in	NN	O	O
the	NN	O	O
RANK	NN	O	B-protein
cytoplasmic	NN	O	I-protein
tail	NN	O	I-protein
.	NN	O	O

These	NN	O	O
TRAF	NN	O	B-protein
binding	NN	O	I-protein
domains	NN	O	I-protein
were	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
functionally	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
RANK	NN	O	B-protein
-dependent	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
NH2-terminal	NN	O	B-protein
kinase	NN	O	I-protein
activities	NN	O	O
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
these	NN	O	O
TRAF	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
exhibited	NN	O	O
selective	NN	O	O
binding	NN	O	O
for	NN	O	O
different	NN	O	O
TRAF	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
TRAF6	NN	O	B-protein
interacted	NN	O	O
with	NN	O	O
membrane-proximal	NN	O	B-protein
determinants	NN	O	I-protein
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
binding	NN	O	O
TRAFs	NN	O	B-protein
1	NN	O	I-protein
,	NN	O	I-protein
2	NN	O	I-protein
,	NN	O	I-protein
3	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
5	NN	O	I-protein
.	NN	O	O

When	NN	O	O
this	NN	O	O
membrane-proximal	NN	O	B-protein
TRAF6	NN	O	I-protein
interaction	NN	O	I-protein
domain	NN	O	I-protein
was	NN	O	O
deleted	NN	O	O
,	NN	O	O
RANK	NN	O	B-protein
-mediated	NN	O	O
NF-kappaB	NN	O	B-protein
signaling	NN	O	O
was	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
while	NN	O	O
c-Jun	NN	O	B-protein
NH2-terminal	NN	O	I-protein
kinase	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
partially	NN	O	O
inhibited	NN	O	O
.	NN	O	O

An	NN	O	O
NH2-terminal	NN	O	B-protein
truncation	NN	O	I-protein
mutant	NN	O	I-protein
of	NN	O	O
TRAF6	NN	O	B-protein
inhibited	NN	O	O
RANKL	NN	O	B-protein
-mediated	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
but	NN	O	O
failed	NN	O	O
to	NN	O	O
affect	NN	O	O
constitutive	NN	O	O
signaling	NN	O	O
induced	NN	O	O
by	NN	O	O
receptor	NN	O	O
overexpression	NN	O	O
,	NN	O	O
revealing	NN	O	O
a	NN	O	O
selective	NN	O	O
role	NN	O	O
for	NN	O	O
TRAF6	NN	O	B-protein
in	NN	O	O
ligand-induced	NN	O	O
activation	NN	O	O
events	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stat6	NN	O	B-protein
inhibits	NN	O	O
human	NN	O	O
interleukin-4	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
differentiation	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
T-helper	NN	O	I-cell_type
(	NN	O	I-cell_type
Th	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
cytokine-secreting	NN	O	B-cell_type
effector	NN	O	I-cell_type
Th	NN	O	I-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
exposure	NN	O	O
to	NN	O	O
multiple	NN	O	O
signals	NN	O	O
,	NN	O	O
including	NN	O	O
exogenous	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

Interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
plays	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
this	NN	O	O
process	NN	O	O
by	NN	O	O
promoting	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
-secreting	NN	O	O
Th2	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
Th2	NN	O	O
cells	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
tightly	NN	O	O
controlled	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
by	NN	O	O
the	NN	O	O
coordinated	NN	O	O
binding	NN	O	O
of	NN	O	O
multiple	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
regulatory	NN	O	O
elements	NN	O	O
in	NN	O	O
the	NN	O	O
proximal	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
(	NN	O	I-protein
NFAT	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
IL-4	NN	O	B-protein
transcription	NN	O	O
and	NN	O	O
interact	NN	O	O
with	NN	O	O
up	NN	O	O
to	NN	O	O
five	NN	O	O
sequences	NN	O	O
(	NN	O	O
termed	NN	O	O
P0	NN	O	O
through	NN	O	O
P4	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
IL-4	NN	O	B-protein
induces	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
are	NN	O	O
not	NN	O	O
known	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
IL-4-activated	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
signal	NN	O	I-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
6	NN	O	I-protein
(	NN	O	O
Stat6	NN	O	B-protein
)	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
this	NN	O	O
effect	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
Stat6	NN	O	B-protein
interacts	NN	O	O
with	NN	O	O
three	NN	O	O
binding	NN	O	O
sites	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

These	NN	O	O
sites	NN	O	O
overlap	NN	O	O
the	NN	O	O
P1	NN	O	B-DNA
,	NN	O	I-DNA
P2	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
P4	NN	O	I-DNA
NFAT	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
in	NN	O	O
regulating	NN	O	O
IL-4	NN	O	B-protein
transcription	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
Stat6	NN	O	B-protein
-deficient	NN	O	O
Jurkat	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
different	NN	O	O
intact	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
cotransfection	NN	O	O
assays	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
multimerized	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
germline	NN	O	B-DNA
IgE	NN	O	I-DNA
promoter	NN	O	I-DNA
was	NN	O	O
highly	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
in	NN	O	O
Stat6-expressing	NN	O	B-cell_type
Jurkat	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
intact	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
was	NN	O	O
repressed	NN	O	O
under	NN	O	O
similar	NN	O	O
conditions	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
is	NN	O	O
highly	NN	O	O
dependent	NN	O	O
on	NN	O	O
promoter	NN	O	O
context	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
factor	NN	O	O
promotes	NN	O	O
IL-4	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
an	NN	O	O
indirect	NN	O	O
manner	NN	O	O
.	NN	O	O

-DOCSTART-	O

Epithelial	NN	O	O
cell-initiated	NN	O	O
inflammation	NN	O	O
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
early	NN	O	O
tissue	NN	O	O
damage	NN	O	O
in	NN	O	O
amebic	NN	O	O
infection	NN	O	O
of	NN	O	O
human	NN	O	O
intestine	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
&	NN	O	O
AIMS	NN	O	O
:	NN	O	O
Entamoeba	NN	O	O
histolytica	NN	O	O
infection	NN	O	O
of	NN	O	O
the	NN	O	O
intestine	NN	O	O
can	NN	O	O
induce	NN	O	O
severe	NN	O	O
gut	NN	O	O
inflammation	NN	O	O
.	NN	O	O

The	NN	O	O
aims	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
were	NN	O	O
to	NN	O	O
assess	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
host	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
in	NN	O	O
the	NN	O	O
tissue	NN	O	O
damage	NN	O	O
observed	NN	O	O
with	NN	O	O
amebiasis	NN	O	O
and	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
intestinal	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cell	NN	O	I-cell_type
in	NN	O	O
initiating	NN	O	O
that	NN	O	O
response	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
E.	NN	O	O
histolytica	NN	O	O
infection	NN	O	O
was	NN	O	O
established	NN	O	O
in	NN	O	O
human	NN	O	O
intestinal	NN	O	O
xenografts	NN	O	O
in	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficient	NN	O	O
(	NN	O	O
SCID-HU-INT	NN	O	O
)	NN	O	O
mice	NN	O	O
.	NN	O	O

Human	NN	O	O
intestinal	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cell	NN	O	I-cell_type
inflammatory	NN	O	O
responses	NN	O	O
to	NN	O	O
amebic	NN	O	O
infection	NN	O	O
were	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
intraluminal	NN	O	O
administration	NN	O	O
of	NN	O	O
an	NN	O	O
antisense	NN	O	O
oligonucleotide	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-protein
p65	NN	O	I-protein
subunit	NN	O	I-protein
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
neutrophils	NN	O	O
in	NN	O	O
tissue	NN	O	O
damage	NN	O	O
observed	NN	O	O
with	NN	O	O
amebiasis	NN	O	O
was	NN	O	O
studied	NN	O	O
by	NN	O	O
depleting	NN	O	O
neutrophils	NN	O	B-cell_type
from	NN	O	O
SCID-HU-INT	NN	O	O
mice	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Administration	NN	O	O
of	NN	O	O
the	NN	O	O
antisense	NN	O	O
oligonucleotide	NN	O	O
blocked	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
human	NN	O	B-protein
interleukin	NN	O	I-protein
1beta	NN	O	I-protein
and	NN	O	O
interleukin	NN	O	B-protein
8	NN	O	I-protein
by	NN	O	O
intestinal	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
inhibited	NN	O	O
neutrophil	NN	O	O
influx	NN	O	O
into	NN	O	O
the	NN	O	O
E.	NN	O	O
histolytica-infected	NN	O	O
intestinal	NN	O	O
xenografts	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
gut	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
by	NN	O	O
the	NN	O	O
antisense	NN	O	O
oligonucleotide	NN	O	O
or	NN	O	O
the	NN	O	O
depletion	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
from	NN	O	O
SCID-HU-	NN	O	O
INT	NN	O	O
mice	NN	O	O
blocked	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
intestinal	NN	O	O
permeability	NN	O	O
observed	NN	O	O
with	NN	O	O
amebic	NN	O	O
infection	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Intestinal	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
initiate	NN	O	O
an	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
with	NN	O	O
resulting	NN	O	O
neutrophil-mediated	NN	O	O
tissue	NN	O	O
damage	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
E.	NN	O	O
histolytica	NN	O	O
infection	NN	O	O
;	NN	O	O
this	NN	O	O
inflammatory	NN	O	O
cascade	NN	O	O
can	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
regulated	NN	O	O
by	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Triptolide	NN	O	O
induces	NN	O	O
apoptotic	NN	O	O
death	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
.	NN	O	O

Extract	NN	O	O
of	NN	O	O
Tripterygium	NN	O	O
wilfordii	NN	O	O
Hook.	NN	O	O
f	NN	O	O
(	NN	O	O
TWHf	NN	O	O
)	NN	O	O
has	NN	O	O
immunosuppressive	NN	O	O
activity	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
as	NN	O	O
anti-inflammatory	NN	O	O
agent	NN	O	O
in	NN	O	O
traditional	NN	O	O
Chinese	NN	O	O
medicine	NN	O	O
for	NN	O	O
centuries	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
triptolide	NN	O	O
is	NN	O	O
the	NN	O	O
major	NN	O	O
active	NN	O	O
component	NN	O	O
in	NN	O	O
the	NN	O	O
extract	NN	O	O
that	NN	O	O
inhibits	NN	O	O
antigen	NN	O	O
or	NN	O	O
mitogen	NN	O	B-protein
-induced	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
attempting	NN	O	O
to	NN	O	O
investigate	NN	O	O
its	NN	O	O
effect	NN	O	O
on	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
found	NN	O	O
triptolide	NN	O	O
induces	NN	O	O
apoptotic	NN	O	O
death	NN	O	O
of	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
and	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
that	NN	O	O
of	NN	O	O
thymocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
triptolide-induced	NN	O	O
apoptosis	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
increase	NN	O	O
of	NN	O	O
DEVD-cleavable	NN	O	O
caspases	NN	O	B-protein
activity	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
caspase	NN	O	B-protein
substrate	NN	O	I-protein
poly	NN	O	I-protein
(	NN	O	I-protein
ADP-ribose	NN	O	I-protein
)	NN	O	I-protein
polymerase	NN	O	I-protein
(	NN	O	O
PARP	NN	O	B-protein
)	NN	O	O
.	NN	O	O

A	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
caspases	NN	O	B-protein
,	NN	O	O
zVAD-FMK	NN	O	O
,	NN	O	O
prevents	NN	O	O
triptolide-induced	NN	O	O
PARP	NN	O	B-protein
degradation	NN	O	O
and	NN	O	O
DNA	NN	O	O
fragmentation	NN	O	O
but	NN	O	O
not	NN	O	O
growth	NN	O	O
arrest	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
enforced	NN	O	O
expression	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
inhibited	NN	O	O
triptolide-induced	NN	O	O
degradation	NN	O	O
of	NN	O	O
PARP	NN	O	B-protein
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
triptolide	NN	O	O
induces	NN	O	O
T	NN	O	O
cell	NN	O	O
apoptosis	NN	O	O
through	NN	O	O
activating	NN	O	O
caspases	NN	O	B-protein
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
the	NN	O	O
growth	NN	O	O
arrest	NN	O	O
and	NN	O	O
apoptotic	NN	O	O
effect	NN	O	O
of	NN	O	O
triptolide	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
activity	NN	O	O
of	NN	O	O
TWHf	NN	O	O
extract	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
regulates	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
vacuolar	NN	O	B-protein
H+-ATPase	NN	O	I-protein
B2	NN	O	I-protein
subunit	NN	O	I-protein
through	NN	O	O
AP-2	NN	O	B-DNA
sites	NN	O	I-DNA
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

During	NN	O	O
monocyte-to-macrophage	NN	O	O
differentiation	NN	O	O
,	NN	O	O
the	NN	O	O
cellular	NN	O	O
content	NN	O	O
of	NN	O	O
vacuolar	NN	O	B-protein
H+-ATPase	NN	O	I-protein
(	NN	O	O
V-ATPase	NN	O	B-protein
)	NN	O	O
increases	NN	O	O
more	NN	O	O
than	NN	O	O
4-fold	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
previously	NN	O	O
that	NN	O	O
amplified	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
B2	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
V-ATPase	NN	O	B-protein
occurs	NN	O	O
solely	NN	O	O
by	NN	O	O
increased	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
5'-untranslated	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
B2	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
containing	NN	O	O
multiple	NN	O	O
consensus	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP-2	NN	O	B-protein
and	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
this	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
AP-2	NN	O	B-DNA
binding	NN	O	I-DNA
sequences	NN	O	I-DNA
are	NN	O	O
essential	NN	O	O
for	NN	O	O
increased	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
B2	NN	O	B-DNA
promoter	NN	O	I-DNA
during	NN	O	O
monocyte-macrophage	NN	O	O
differentiation	NN	O	O
and	NN	O	O
that	NN	O	O
AP-2	NN	O	B-protein
,	NN	O	O
expressed	NN	O	O
exogenously	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
and	NN	O	O
other	NN	O	O
cells	NN	O	O
,	NN	O	O
activates	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
B2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
from	NN	O	O
THP-1	NN	O	B-cell_line
and	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
identified	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
several	NN	O	O
AP-2	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
B2	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
does	NN	O	O
not	NN	O	O
react	NN	O	O
with	NN	O	O
AP-2	NN	O	B-protein
antibodies	NN	O	I-protein
,	NN	O	O
and	NN	O	O
has	NN	O	O
a	NN	O	O
DNA	NN	O	B-DNA
sequence	NN	O	I-DNA
binding	NN	O	O
affinity	NN	O	O
profile	NN	O	O
that	NN	O	O
differs	NN	O	O
from	NN	O	O
AP-2	NN	O	B-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
novel	NN	O	O
AP-2-like	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
V-ATPase	NN	O	B-protein
B	NN	O	I-protein
subunit	NN	O	I-protein
amplification	NN	O	O
during	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

c-Jun	NN	O	O
and	NN	O	O
GST-pi	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
plasma	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Bone	NN	O	O
marrow	NN	O	O
samples	NN	O	O
from	NN	O	O
33	NN	O	O
patients	NN	O	O
affected	NN	O	O
by	NN	O	O
MM	NN	O	O
and	NN	O	O
MGUS	NN	O	O
,	NN	O	O
and	NN	O	O
8	NN	O	O
patients	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
lymphoproliferative	NN	O	O
diseases	NN	O	O
were	NN	O	O
studied	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
(	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
glutathione-S-transferase	NN	O	B-protein
pi	NN	O	I-protein
(	NN	O	O
GST-pi	NN	O	B-protein
)	NN	O	O
using	NN	O	O
immunocytochemical	NN	O	O
methods	NN	O	O
.	NN	O	O

A	NN	O	O
high	NN	O	O
and	NN	O	O
frequent	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
proteins	NN	O	O
was	NN	O	O
found	NN	O	O
both	NN	O	O
in	NN	O	O
MM	NN	O	O
and	NN	O	O
MGUS	NN	O	O
patients	NN	O	O
(	NN	O	O
31/33	NN	O	O
patients	NN	O	O
positive	NN	O	O
for	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
29/33	NN	O	O
patients	NN	O	O
positive	NN	O	O
for	NN	O	O
GST-pi	NN	O	B-protein
)	NN	O	O
and	NN	O	O
in	NN	O	O
controls	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
monoclonal	NN	O	O
gammopathy	NN	O	O
(	NN	O	O
7/8	NN	O	O
patients	NN	O	O
positive	NN	O	O
for	NN	O	O
both	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
GST-pi	NN	O	B-protein
)	NN	O	O
.	NN	O	O

No	NN	O	O
statistically	NN	O	O
significant	NN	O	O
correlation	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
c-Jun-	NN	O	B-cell_line
and	NN	O	I-cell_line
GST-pi-positive	NN	O	I-cell_line
plasma	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
proteins	NN	O	O
was	NN	O	O
not	NN	O	O
related	NN	O	O
to	NN	O	O
clinical	NN	O	O
or	NN	O	O
laboratory	NN	O	O
data	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
seem	NN	O	O
to	NN	O	O
confirm	NN	O	O
a	NN	O	O
possible	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
complex	NN	O	I-protein
AP-1	NN	O	B-protein
in	NN	O	O
activating	NN	O	O
GST-pi	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
plasma	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
and	NN	O	O
prevention	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
eosinophils	NN	O	B-cell_type
by	NN	O	O
cytokines	NN	O	B-protein
such	NN	O	O
as	NN	O	O
IL-5	NN	O	B-protein
and	NN	O	O
GM-CSF	NN	O	B-protein
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
allergic	NN	O	O
and	NN	O	O
parasitic	NN	O	O
disorders	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
there	NN	O	O
has	NN	O	O
been	NN	O	O
some	NN	O	O
progress	NN	O	O
in	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
activated	NN	O	O
by	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
IL-3	NN	O	B-protein
,	NN	O	O
IL-5	NN	O	B-protein
and	NN	O	O
GM-CSF	NN	O	B-protein
receptors	NN	O	I-protein
share	NN	O	O
a	NN	O	O
common	NN	O	B-protein
signal	NN	O	I-protein
transducer	NN	O	I-protein
that	NN	O	O
possesses	NN	O	O
no	NN	O	O
intrinsic	NN	O	B-protein
kinase	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
that	NN	O	O
eosinophil	NN	O	O
stimulation	NN	O	O
by	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
is	NN	O	O
associated	NN	O	O
with	NN	O	O
increases	NN	O	O
in	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
several	NN	O	O
cellular	NN	O	O
substrates	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
past	NN	O	O
few	NN	O	O
years	NN	O	O
,	NN	O	O
there	NN	O	O
has	NN	O	O
been	NN	O	O
some	NN	O	O
progress	NN	O	O
in	NN	O	O
defining	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
kinases	NN	O	O
that	NN	O	O
are	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
IL-3/IL-5/GM-CSF	NN	O	B-protein
receptor	NN	O	I-protein
beta-subunit	NN	O	I-protein
in	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

This	NN	O	O
review	NN	O	O
will	NN	O	O
concentrate	NN	O	O
on	NN	O	O
this	NN	O	O
topic	NN	O	O
and	NN	O	O
on	NN	O	O
its	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
eosinophil	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Requirement	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
p45	NN	O	B-protein
NF-E2	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
butyric	NN	O	O
acid-induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Butyric	NN	O	O
acid	NN	O	O
(	NN	O	O
BA	NN	O	O
)	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
induce	NN	O	O
overexpression	NN	O	O
of	NN	O	O
fetal	NN	O	B-protein
hemoglobin	NN	O	I-protein
and	NN	O	O
then	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
BA	NN	O	O
is	NN	O	O
currently	NN	O	O
under	NN	O	O
clinical	NN	O	O
investigation	NN	O	O
as	NN	O	O
a	NN	O	O
potential	NN	O	O
therapy	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
sickle	NN	O	O
cell	NN	O	O
disease	NN	O	O
and	NN	O	O
cancer	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
BA-induced	NN	O	O
differentiation	NN	O	O
remain	NN	O	O
largely	NN	O	O
unknown	NN	O	O
.	NN	O	O

Previous	NN	O	O
reports	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
BA-induced	NN	O	O
overexpression	NN	O	O
of	NN	O	O
erythroid	NN	O	B-DNA
genes	NN	O	I-DNA
occurred	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
erythroid	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
intend	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
requirement	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
NF-E2	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
BA-induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
K562	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Time-course	NN	O	O
experiments	NN	O	O
showed	NN	O	O
that	NN	O	O
nuclear	NN	O	O
levels	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
p45	NN	O	B-protein
NF-E2	NN	O	I-protein
proteins	NN	O	O
increased	NN	O	O
during	NN	O	O
BA	NN	O	O
treatment	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
antisense	NN	O	O
oligodeoxynucleotides	NN	O	O
targeting	NN	O	O
either	NN	O	O
GATA-1	NN	O	B-protein
or	NN	O	O
p45	NN	O	B-protein
NF-E2	NN	O	I-protein
proteins	NN	O	O
inhibited	NN	O	O
both	NN	O	O
protein	NN	O	O
expression	NN	O	O
and	NN	O	O
BA-induced	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
BA-induced	NN	O	O
cell	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
was	NN	O	O
not	NN	O	O
affected	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
the	NN	O	O
first	NN	O	O
direct	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
requirement	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
NF-E2	NN	O	B-protein
in	NN	O	O
BA-induced	NN	O	O
differentiation	NN	O	O
process	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterisation	NN	O	O
of	NN	O	O
regulatory	NN	O	O
sequences	NN	O	O
at	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-DNA
virus	NN	O	I-DNA
BamHI	NN	O	I-DNA
W	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Epstein-Barr	NN	O	O
virus	NN	O	O
,	NN	O	O
a	NN	O	O
human	NN	O	O
gammaherpesvirus	NN	O	O
,	NN	O	O
possesses	NN	O	O
a	NN	O	O
unique	NN	O	O
set	NN	O	O
of	NN	O	O
latent	NN	O	B-DNA
genes	NN	O	I-DNA
whose	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
leads	NN	O	O
to	NN	O	O
cell	NN	O	O
growth	NN	O	O
transformation	NN	O	O
.	NN	O	O

The	NN	O	O
initiation	NN	O	O
of	NN	O	O
this	NN	O	O
growth	NN	O	O
transforming	NN	O	O
infection	NN	O	O
depends	NN	O	O
on	NN	O	O
a	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
BamHI	NN	O	O
W	NN	O	O
(	NN	O	O
Wp	NN	O	O
)	NN	O	O
whose	NN	O	O
regulation	NN	O	O
is	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

Using	NN	O	O
Wp	NN	O	B-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
in	NN	O	O
vitro	NN	O	O
transfection	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
Wp	NN	O	O
was	NN	O	O
11-	NN	O	O
to	NN	O	O
190-fold	NN	O	O
more	NN	O	O
active	NN	O	O
in	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
than	NN	O	O
in	NN	O	O
non-B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
that	NN	O	O
three	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
(	NN	O	O
termed	NN	O	O
UAS1	NN	O	B-DNA
,	NN	O	O
UAS2	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
UAS3	NN	O	B-DNA
)	NN	O	O
contributed	NN	O	O
to	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
upstream	NN	O	O
regions	NN	O	O
UAS3	NN	O	B-DNA
(	NN	O	O
-1168	NN	O	B-DNA
to	NN	O	I-DNA
-440	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
UAS2	NN	O	B-DNA
(	NN	O	O
-352	NN	O	B-DNA
to	NN	O	I-DNA
-264	NN	O	I-DNA
)	NN	O	O
both	NN	O	O
functioned	NN	O	O
in	NN	O	O
a	NN	O	O
cell	NN	O	O
lineage-independent	NN	O	O
manner	NN	O	O
and	NN	O	O
were	NN	O	O
together	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
bulk	NN	O	O
of	NN	O	O
Wp	NN	O	O
activity	NN	O	O
in	NN	O	O
non-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
mutational	NN	O	O
analysis	NN	O	O
indicated	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
a	NN	O	O
YY1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
UAS2	NN	O	B-DNA
in	NN	O	O
that	NN	O	O
context	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
UAS1	NN	O	B-DNA
(	NN	O	O
-140	NN	O	B-DNA
to	NN	O	I-DNA
-87	NN	O	I-DNA
)	NN	O	O
was	NN	O	O
B	NN	O	O
cell	NN	O	O
specific	NN	O	O
and	NN	O	O
was	NN	O	O
the	NN	O	O
key	NN	O	O
determinant	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
's	NN	O	O
increased	NN	O	O
activity	NN	O	O
in	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Mutational	NN	O	O
analysis	NN	O	O
of	NN	O	O
UAS1	NN	O	B-DNA
sequences	NN	O	I-DNA
combined	NN	O	O
with	NN	O	O
in	NN	O	O
vitro	NN	O	O
bandshift	NN	O	O
assays	NN	O	O
revealed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
three	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
this	NN	O	O
region	NN	O	O
.	NN	O	O

When	NN	O	O
mutations	NN	O	O
that	NN	O	O
abolished	NN	O	O
factor	NN	O	O
binding	NN	O	O
in	NN	O	O
bandshift	NN	O	O
assays	NN	O	O
were	NN	O	O
introduced	NN	O	O
into	NN	O	O
a	NN	O	O
Wp	NN	O	B-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
any	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
UAS1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
was	NN	O	O
sufficient	NN	O	O
to	NN	O	O
reduce	NN	O	O
promoter	NN	O	O
activity	NN	O	O
by	NN	O	O
10-	NN	O	O
to	NN	O	O
30-fold	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

From	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
,	NN	O	O
two	NN	O	O
of	NN	O	O
these	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
novel	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
third	NN	O	O
was	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
cyclic	NN	O	B-DNA
AMP	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CRE	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
this	NN	O	O
CRE	NN	O	B-DNA
interacts	NN	O	O
with	NN	O	O
CREB	NN	O	B-protein
and	NN	O	I-protein
ATF1	NN	O	I-protein
proteins	NN	O	I-protein
present	NN	O	O
in	NN	O	O
B	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
interaction	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
Wp	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
situ	NN	O	O
RT-PCR	NN	O	O
detection	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	B-RNA
virus	NN	O	I-RNA
immediate-early	NN	O	I-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
CD4+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

AIDS-related	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
-associated	NN	O	O
T	NN	O	O
cell	NN	O	O
lymphomas	NN	O	O
are	NN	O	O
emerging	NN	O	O
as	NN	O	O
a	NN	O	O
new	NN	O	O
,	NN	O	O
distinct	NN	O	O
histopathological	NN	O	O
entity	NN	O	O
.	NN	O	O

The	NN	O	O
pathway	NN	O	O
whereby	NN	O	O
EBV	NN	O	O
infects	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
initial	NN	O	O
EBV	NN	O	O
transcriptional	NN	O	O
program	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
not	NN	O	O
been	NN	O	O
established	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
shed	NN	O	O
light	NN	O	O
on	NN	O	O
the	NN	O	O
early	NN	O	O
events	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	O
infection	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
have	NN	O	O
used	NN	O	O
in	NN	O	O
situ	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
based	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
initial	NN	O	O
EBV	NN	O	O
transcriptional	NN	O	O
program	NN	O	O
in	NN	O	O
homogeneous	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
and	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Following	NN	O	O
EBV	NN	O	O
infection	NN	O	O
,	NN	O	O
Epstein-Barr	NN	O	O
nuclear	NN	O	O
antigen	NN	O	O
(	NN	O	O
EBNA	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
could	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
rosetting	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
and	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Only	NN	O	O
a	NN	O	O
few	NN	O	O
cells	NN	O	O
showed	NN	O	O
viral	NN	O	B-protein
capsid	NN	O	I-protein
antigen	NN	O	I-protein
(	NN	O	O
VCA	NN	O	B-protein
)	NN	O	O
.	NN	O	O

EBV	NN	O	B-RNA
immediate-early	NN	O	I-RNA
gene	NN	O	I-RNA
transcripts	NN	O	I-RNA
(	NN	O	O
BZLF1	NN	O	B-RNA
,	NN	O	O
BRLF1	NN	O	B-RNA
,	NN	O	O
and	NN	O	O
BMLF1	NN	O	B-RNA
)	NN	O	O
encoded	NN	O	O
in	NN	O	O
the	NN	O	O
BamHI	NN	O	O
Z	NN	O	O
,	NN	O	O
R	NN	O	O
,	NN	O	O
and	NN	O	O
M	NN	O	O
fragments	NN	O	O
could	NN	O	O
be	NN	O	O
detected	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
RT-PCR	NN	O	O
in	NN	O	O
the	NN	O	O
EBV	NN	O	O
producer	NN	O	O
cell	NN	O	O
line	NN	O	O
B95.8	NN	O	O
.	NN	O	O

Both	NN	O	O
BZLF1	NN	O	B-RNA
and	NN	O	I-RNA
BRLF1	NN	O	I-RNA
immediate-early	NN	O	I-RNA
transcripts	NN	O	I-RNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
BMLF1	NN	O	B-RNA
transcript	NN	O	I-RNA
,	NN	O	O
could	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
individual	NN	O	O
CD4+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
infected	NN	O	O
with	NN	O	O
EBV	NN	O	O
.	NN	O	O

Demonstration	NN	O	O
of	NN	O	O
EBV	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcripts	NN	O	I-RNA
encoding	NN	O	O
immediate-early	NN	O	B-protein
transcriptional	NN	O	I-protein
transactivators	NN	O	I-protein
in	NN	O	O
EBV-infected	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
provides	NN	O	O
the	NN	O	O
first	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
possible	NN	O	O
mechanism	NN	O	O
whereby	NN	O	O
EBV	NN	O	O
could	NN	O	O
contribute	NN	O	O
to	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
EBV-associated	NN	O	O
T	NN	O	O
cell	NN	O	O
malignancies	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
linkage	NN	O	O
between	NN	O	O
T-cell	NN	O	O
and	NN	O	O
dendritic	NN	O	O
cell	NN	O	O
development	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
thymus	NN	O	O
.	NN	O	O

Thymic	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
DC	NN	O	B-cell_type
)	NN	O	O
mediate	NN	O	O
negative	NN	O	O
selection	NN	O	O
at	NN	O	O
a	NN	O	O
relatively	NN	O	O
late	NN	O	O
stage	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	O
developmental	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
thymic	NN	O	B-cell_type
DC	NN	O	I-cell_type
and	NN	O	O
of	NN	O	O
T-lineage	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
linked	NN	O	O
via	NN	O	O
a	NN	O	O
common	NN	O	O
precursor	NN	O	O
at	NN	O	O
an	NN	O	O
early	NN	O	O
stage	NN	O	O
of	NN	O	O
thymocyte	NN	O	O
development	NN	O	O
.	NN	O	O

T-lineage	NN	O	O
precursor	NN	O	O
populations	NN	O	O
from	NN	O	O
the	NN	O	O
adult	NN	O	O
mouse	NN	O	O
thymus	NN	O	O
,	NN	O	O
prior	NN	O	O
to	NN	O	O
T-cell	NN	O	O
receptor	NN	O	O
gene	NN	O	O
rearrangement	NN	O	O
,	NN	O	O
display	NN	O	O
a	NN	O	O
capacity	NN	O	O
to	NN	O	O
produce	NN	O	O
DC	NN	O	B-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
,	NN	O	O
and	NN	O	O
are	NN	O	O
very	NN	O	O
efficient	NN	O	O
precursors	NN	O	O
of	NN	O	O
DC	NN	O	B-cell_type
in	NN	O	O
culture	NN	O	O
.	NN	O	O

These	NN	O	O
lymphoid/DC	NN	O	B-cell_type
precursors	NN	O	I-cell_type
have	NN	O	O
little	NN	O	O
capacity	NN	O	O
to	NN	O	O
form	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
thymic	NN	O	B-cell_type
DC	NN	O	I-cell_type
are	NN	O	O
a	NN	O	O
lymphoid-related	NN	O	O
rather	NN	O	O
than	NN	O	O
myeloid-related	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
myeloid-related	NN	O	B-cell_type
DC	NN	O	I-cell_type
,	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
these	NN	O	O
lymphoid-related	NN	O	B-cell_type
DC	NN	O	I-cell_type
in	NN	O	O
vivo	NN	O	O
or	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

DC	NN	O	B-cell_type
can	NN	O	O
develop	NN	O	O
in	NN	O	O
mutant	NN	O	O
mice	NN	O	O
lacking	NN	O	O
mature	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
provided	NN	O	O
the	NN	O	O
common	NN	O	O
precursors	NN	O	O
are	NN	O	O
present	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
mutant	NN	O	O
mice	NN	O	O
lacking	NN	O	O
functional	NN	O	O
Ikaros	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
there	NN	O	O
are	NN	O	O
deficiencies	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
DC	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Host	NN	O	O
control	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
parasitism	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
.	NN	O	O

Post	NN	O	O
HIV-1	NN	O	O
entry	NN	O	O
,	NN	O	O
productive	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
overcoming	NN	O	O
several	NN	O	O
cellular	NN	O	O
blocks	NN	O	O
to	NN	O	O
provirus	NN	O	O
establishment	NN	O	O
and	NN	O	O
replication	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
unknown	NN	O	O
host	NN	O	O
intracellular	NN	O	O
events	NN	O	O
overcomes	NN	O	O
such	NN	O	O
inhibitory	NN	O	O
steps	NN	O	O
and	NN	O	O
is	NN	O	O
concomitant	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NFATc	NN	O	B-protein
was	NN	O	O
sufficient	NN	O	O
as	NN	O	O
a	NN	O	O
cellular	NN	O	O
factor	NN	O	O
to	NN	O	O
induce	NN	O	O
a	NN	O	O
highly	NN	O	O
permissive	NN	O	O
state	NN	O	O
for	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NFATc	NN	O	B-protein
overcame	NN	O	O
a	NN	O	O
blockade	NN	O	O
at	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
and	NN	O	O
permitted	NN	O	O
active	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

Pharmacologic	NN	O	O
blockade	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
NFAT	NN	O	I-protein
activity	NN	O	O
by	NN	O	O
FK506	NN	O	O
or	NN	O	O
CsA	NN	O	O
inhibited	NN	O	O
synthesis	NN	O	O
of	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
and	NN	O	O
also	NN	O	O
potently	NN	O	O
blocked	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
therefore	NN	O	O
can	NN	O	O
become	NN	O	O
competent	NN	O	O
for	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
by	NN	O	O
control	NN	O	O
of	NN	O	O
regulated	NN	O	B-protein
host	NN	O	I-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
the	NN	O	O
NFATc	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
host	NN	O	O
mechanisms	NN	O	O
regulated	NN	O	O
by	NN	O	O
such	NN	O	O
permissivity	NN	O	O
factors	NN	O	O
are	NN	O	O
potential	NN	O	O
targets	NN	O	O
for	NN	O	O
anti-HIV-1	NN	O	O
therapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT1	NN	O	B-protein
nuclear	NN	O	O
import	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
noninvasive	NN	O	O
delivery	NN	O	O
of	NN	O	O
peptide	NN	O	O
carrying	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
localization	NN	O	I-protein
sequence	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	I-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
Ags	NN	O	B-protein
or	NN	O	O
cytokines	NN	O	B-protein
results	NN	O	O
in	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT	NN	O	B-protein
from	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
import	NN	O	O
process	NN	O	O
is	NN	O	O
recognition	NN	O	O
of	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
localization	NN	O	I-protein
sequence	NN	O	I-protein
(	NN	O	O
NLS	NN	O	B-protein
)	NN	O	O
within	NN	O	O
the	NN	O	O
karyophilic	NN	O	B-protein
protein	NN	O	I-protein
by	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	B-protein
receptor	NN	O	I-protein
such	NN	O	O
as	NN	O	O
the	NN	O	O
importin	NN	O	B-protein
(	NN	O	I-protein
karyopherin	NN	O	I-protein
)	NN	O	I-protein
-alpha	NN	O	I-protein
subunit	NN	O	I-protein
.	NN	O	O

The	NN	O	O
NLSs	NN	O	B-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NFAT	NN	O	B-protein
differ	NN	O	O
and	NN	O	O
the	NN	O	O
NLS	NN	O	B-protein
of	NN	O	O
STAT1	NN	O	B-protein
has	NN	O	O
not	NN	O	O
yet	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

Herein	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
inducible	NN	O	O
nuclear	NN	O	O
import	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT1	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
is	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
cell-permeable	NN	O	O
peptide	NN	O	O
carrying	NN	O	O
the	NN	O	O
NLS	NN	O	B-protein
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	I-protein
subunit	NN	O	I-protein
.	NN	O	O

NLS	NN	O	B-protein
peptide-mediated	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
import	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
results	NN	O	O
in	NN	O	O
inhibition	NN	O	O
of	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
alpha	NN	O	O
and	NN	O	O
IL-2	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
processes	NN	O	O
dependent	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
or	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

Further	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
inhibitory	NN	O	O
NLS	NN	O	B-protein
peptide	NN	O	O
interacts	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	B-protein
NLS	NN	O	I-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
comprised	NN	O	O
of	NN	O	O
the	NN	O	O
Rch1/importin	NN	O	B-protein
(	NN	O	I-protein
karyopherin	NN	O	I-protein
)	NN	O	I-protein
-beta	NN	O	I-protein
heterodimer	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
inducible	NN	O	O
nuclear	NN	O	O
import	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT1	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
can	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
NLS	NN	O	B-protein
peptide	NN	O	O
delivered	NN	O	O
noninvasively	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
target	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
importin	NN	O	B-protein
(	NN	O	I-protein
karyopherin	NN	O	I-protein
)	NN	O	I-protein
-alpha	NN	O	I-protein
beta	NN	O	I-protein
NLS	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Membrane-associated	NN	O	B-protein
lymphotoxin	NN	O	I-protein
on	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
activates	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
an	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
pathway	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Inhibition	NN	O	O
of	NN	O	O
complement	NN	O	O
in	NN	O	O
small	NN	O	O
animal	NN	O	O
models	NN	O	O
of	NN	O	O
xenotransplantation	NN	O	O
has	NN	O	O
demonstrated	NN	O	O
graft	NN	O	O
infiltration	NN	O	O
with	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
NK	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	O
associated	NN	O	O
with	NN	O	O
endothelial	NN	O	B-cell_type
cell	NN	O	I-cell_type
(	NN	O	O
EC	NN	O	B-cell_type
)	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
activate	NN	O	O
porcine	NN	O	O
EC	NN	O	B-cell_type
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
the	NN	O	O
NK	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
NK92	NN	O	I-cell_line
to	NN	O	O
define	NN	O	O
the	NN	O	O
molecular	NN	O	O
and	NN	O	O
cellular	NN	O	O
basis	NN	O	O
of	NN	O	O
NK	NN	O	O
cell-mediated	NN	O	O
EC	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
EC	NN	O	B-cell_type
were	NN	O	O
transfected	NN	O	O
with	NN	O	O
either	NN	O	O
reporter	NN	O	O
constructs	NN	O	O
containing	NN	O	O
the	NN	O	O
luciferase	NN	O	B-DNA
gene	NN	O	I-DNA
driven	NN	O	O
either	NN	O	O
by	NN	O	O
E-selectin	NN	O	B-DNA
or	NN	O	I-DNA
interleukin	NN	O	I-DNA
(	NN	O	I-DNA
IL	NN	O	I-DNA
)	NN	O	I-DNA
-8	NN	O	I-DNA
promoters	NN	O	I-DNA
or	NN	O	O
a	NN	O	O
synthetic	NN	O	O
NF-kappaB-dependent	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
a	NN	O	O
dominant-negative	NN	O	B-DNA
mutant	NN	O	I-DNA
tumor	NN	O	I-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
receptor	NN	O	I-DNA
I	NN	O	I-DNA
(	NN	O	I-DNA
TNFRI	NN	O	I-DNA
)	NN	O	I-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
was	NN	O	O
co-transfected	NN	O	O
in	NN	O	O
inhibition	NN	O	O
studies	NN	O	O
.	NN	O	O

Forty-eight	NN	O	O
hours	NN	O	O
after	NN	O	O
transfection	NN	O	O
,	NN	O	O
EC	NN	O	B-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
membrane	NN	O	I-cell_type
extracts	NN	O	I-cell_type
for	NN	O	O
7	NN	O	O
hr	NN	O	O
and	NN	O	O
activation	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
a	NN	O	O
luciferase	NN	O	B-protein
assay	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Co-culture	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
transfected	NN	O	O
EC	NN	O	B-cell_type
enhanced	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

NK	NN	O	O
cell	NN	O	O
membrane	NN	O	O
extracts	NN	O	O
retained	NN	O	O
the	NN	O	O
capacity	NN	O	O
to	NN	O	O
activate	NN	O	O
EC	NN	O	B-cell_type
and	NN	O	O
induced	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
(	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Western	NN	O	O
blotting	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
and	NN	O	O
membrane	NN	O	O
extracts	NN	O	O
detected	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Lymphotoxin-alpha	NN	O	B-protein
(	NN	O	O
LTalpha	NN	O	B-protein
)	NN	O	O
but	NN	O	O
not	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
LTalpha	NN	O	B-protein
was	NN	O	O
secreted	NN	O	O
in	NN	O	O
NK	NN	O	B-cell_line
:	NN	O	I-cell_line
EC	NN	O	I-cell_line
co-cultures	NN	O	I-cell_line
.	NN	O	O

Co-transfection	NN	O	O
with	NN	O	O
dominant-negative	NN	O	B-protein
mutant	NN	O	I-protein
TNFRI	NN	O	I-protein
inhibited	NN	O	O
EC	NN	O	B-cell_type
activation	NN	O	O
by	NN	O	O
NK	NN	O	O
cell	NN	O	O
membrane	NN	O	O
extracts	NN	O	O
and	NN	O	O
by	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
80	NN	O	O
%	NN	O	O
and	NN	O	O
47	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
pattern	NN	O	O
of	NN	O	O
inhibition	NN	O	O
was	NN	O	O
observed	NN	O	O
using	NN	O	O
anti-human	NN	O	O
LT	NN	O	O
sera	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Human	NN	O	O
NK	NN	O	O
cell	NN	O	O
membrane-bound	NN	O	O
LT	NN	O	O
signals	NN	O	O
across	NN	O	O
species	NN	O	O
via	NN	O	O
TNFRI	NN	O	B-protein
,	NN	O	O
leading	NN	O	O
to	NN	O	O
NF-kappaB	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
transcription	NN	O	O
of	NN	O	O
E-selectin	NN	O	B-protein
and	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
which	NN	O	O
results	NN	O	O
in	NN	O	O
EC	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

The	NN	O	O
discrepancy	NN	O	O
in	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
inhibition	NN	O	O
by	NN	O	O
membrane	NN	O	O
extracts	NN	O	O
and	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
mutant	NN	O	B-protein
TNFRI	NN	O	I-protein
suggests	NN	O	O
that	NN	O	O
additional	NN	O	O
pathways	NN	O	O
are	NN	O	O
utilized	NN	O	O
by	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
activate	NN	O	O
EC	NN	O	B-cell_type

-DOCSTART-	O

Ras	NN	O	B-protein
-dependent	NN	O	O
,	NN	O	O
Ca2+-stimulated	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
by	NN	O	O
a	NN	O	O
constitutively	NN	O	O
active	NN	O	O
Cbl	NN	O	B-protein
mutant	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
stimulation	NN	O	O
induces	NN	O	O
rapid	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
including	NN	O	O
Cbl	NN	O	B-protein
,	NN	O	O
a	NN	O	O
protooncogene	NN	O	B-protein
product	NN	O	I-protein
whose	NN	O	O
function	NN	O	O
remains	NN	O	O
unclear	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
first	NN	O	O
step	NN	O	O
toward	NN	O	O
elucidating	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
Cbl	NN	O	B-protein
in	NN	O	O
TCR	NN	O	B-protein
-initiated	NN	O	O
signaling	NN	O	O
,	NN	O	O
we	NN	O	O
evaluated	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
wild-type	NN	O	B-protein
Cbl	NN	O	I-protein
or	NN	O	O
a	NN	O	O
transforming	NN	O	O
Cbl	NN	O	B-protein
mutant	NN	O	I-protein
(	NN	O	O
70Z/3	NN	O	O
)	NN	O	O
to	NN	O	O
induce	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
of	NN	O	I-DNA
activated	NN	O	I-DNA
T	NN	O	I-DNA
cells	NN	O	I-DNA
(	NN	O	I-DNA
NFAT	NN	O	I-DNA
)	NN	O	I-DNA
element	NN	O	I-DNA
derived	NN	O	O
from	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
(	NN	O	I-DNA
IL2	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
transiently	NN	O	O
cotransfected	NN	O	O
Jurkat-TAg	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

70Z/3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Cbl	NN	O	B-protein
,	NN	O	O
caused	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
which	NN	O	O
was	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
drastically	NN	O	O
reduced	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
pretreatment	NN	O	O
.	NN	O	O

A	NN	O	O
point	NN	O	O
mutation	NN	O	O
of	NN	O	O
a	NN	O	O
potential	NN	O	O
phosphatidylinositol	NN	O	B-DNA
3-kinase	NN	O	I-DNA
(	NN	O	I-DNA
PI3-K	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
Y731EAM	NN	O	B-DNA
to	NN	O	O
Y731EAC	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
70Z/3	NN	O	B-protein
disrupted	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
PI3-K	NN	O	B-protein
with	NN	O	O
70Z/3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
reduce	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
interaction	NN	O	O
between	NN	O	O
Cbl	NN	O	B-protein
and	NN	O	O
PI3-K	NN	O	B-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
in	NN	O	O
the	NN	O	O
70Z/3	NN	O	B-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

Additional	NN	O	O
mapping	NN	O	O
studies	NN	O	O
indicated	NN	O	O
that	NN	O	O
defined	NN	O	O
deletions	NN	O	O
of	NN	O	O
C-terminal	NN	O	O
70Z/3	NN	O	B-protein
sequences	NN	O	O
affected	NN	O	O
to	NN	O	O
a	NN	O	O
variable	NN	O	O
degree	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
stimulate	NN	O	O
NFAT	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Strikingly	NN	O	O
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
346	NN	O	B-protein
C-terminal	NN	O	I-protein
residues	NN	O	I-protein
augmented	NN	O	O
this	NN	O	O
activity	NN	O	O
,	NN	O	O
whereas	NN	O	O
removal	NN	O	O
of	NN	O	O
20	NN	O	O
additional	NN	O	O
residues	NN	O	O
abolished	NN	O	O
it	NN	O	O
.	NN	O	O

Coexpression	NN	O	O
of	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
Ras	NN	O	I-protein
abrogated	NN	O	O
the	NN	O	O
basal	NN	O	O
or	NN	O	O
ionomycin-stimulated	NN	O	O
,	NN	O	O
70Z/3	NN	O	B-protein
-mediated	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
functional	NN	O	B-protein
Ras	NN	O	I-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
this	NN	O	O
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
implicate	NN	O	O
Cbl	NN	O	B-protein
in	NN	O	O
Ras	NN	O	B-protein
-dependent	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
which	NN	O	O
lead	NN	O	O
to	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Requirement	NN	O	O
of	NN	O	O
prestimulated	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
for	NN	O	O
endothelial	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

Involvement	NN	O	O
of	NN	O	O
TNF	NN	O	O
alpha	NN	O	O
.	NN	O	O

Blood	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
spontaneously	NN	O	O
activate	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
culture	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
adhesion	NN	O	O
of	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
onto	NN	O	O
the	NN	O	O
endothelial	NN	O	O
surface	NN	O	O
and	NN	O	O
overproduction	NN	O	O
of	NN	O	O
endothelial	NN	O	O
proteins	NN	O	O
such	NN	O	O
as	NN	O	O
von	NN	O	B-protein
Willebrand	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
vWf	NN	O	B-protein
)	NN	O	O
and	NN	O	O
plasminogen	NN	O	B-protein
activator	NN	O	I-protein
inhibitor	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
PAI-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

To	NN	O	O
overcome	NN	O	O
the	NN	O	O
difficulty	NN	O	O
in	NN	O	O
obtaining	NN	O	O
quiescent	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
promonocytic	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
activate	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-prestimulated	NN	O	O
and	NN	O	O
untreated	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
cocultured	NN	O	O
with	NN	O	O
resting	NN	O	B-cell_type
human	NN	O	I-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
HUVEC	NN	O	B-cell_type
)	NN	O	O
for	NN	O	O
3	NN	O	O
and	NN	O	O
24	NN	O	O
h	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
colimycin	NN	O	O
to	NN	O	O
neutralize	NN	O	O
LPS	NN	O	O
traces	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
untreated	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
had	NN	O	O
little	NN	O	O
effect	NN	O	O
on	NN	O	O
HUVEC	NN	O	B-cell_type
adhesiveness	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
prestimulated	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
followed	NN	O	O
by	NN	O	O
a	NN	O	O
noticeable	NN	O	O
adhesion	NN	O	O
after	NN	O	O
3	NN	O	O
h	NN	O	O
which	NN	O	O
reversed	NN	O	O
to	NN	O	O
basal	NN	O	O
values	NN	O	O
within	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

Under	NN	O	O
these	NN	O	O
conditions	NN	O	O
HUVEC	NN	O	B-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
VCAM-1	NN	O	B-protein
and	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
were	NN	O	O
increased	NN	O	O
at	NN	O	O
3	NN	O	O
h	NN	O	O
with	NN	O	O
only	NN	O	O
ICAM-1	NN	O	B-protein
remaining	NN	O	O
overexpressed	NN	O	O
at	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

Diffusible	NN	O	B-protein
endothelial	NN	O	I-protein
proteins	NN	O	I-protein
such	NN	O	O
as	NN	O	O
soluble	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
PAI-1	NN	O	B-protein
and	NN	O	O
vWf	NN	O	B-protein
to	NN	O	O
a	NN	O	O
minimal	NN	O	O
extent	NN	O	O
,	NN	O	O
increased	NN	O	O
in	NN	O	O
supernatants	NN	O	O
from	NN	O	O
HUVEC	NN	O	B-cell_type
cocultured	NN	O	O
for	NN	O	O
24	NN	O	O
h	NN	O	O
with	NN	O	O
prestimulated	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
those	NN	O	O
cocultures	NN	O	O
,	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
concentrations	NN	O	O
peaked	NN	O	O
at	NN	O	O
3	NN	O	O
h	NN	O	O
whereas	NN	O	O
IL-1	NN	O	O
beta	NN	O	O
levels	NN	O	O
progressively	NN	O	O
rose	NN	O	O
until	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
an	NN	O	O
anti-TNF	NN	O	B-protein
alpha	NN	O	I-protein
antibody	NN	O	I-protein
decreased	NN	O	O
by	NN	O	O
40	NN	O	O
%	NN	O	O
E-selectin	NN	O	B-protein
and	NN	O	O
ICAM-1	NN	O	B-protein
induction	NN	O	O
and	NN	O	O
suppressed	NN	O	O
PAI-1	NN	O	B-protein
overproduction	NN	O	O
with	NN	O	O
a	NN	O	O
weak	NN	O	O
effect	NN	O	O
on	NN	O	O
vWf	NN	O	B-protein
.	NN	O	O

An	NN	O	O
anti-IL-1	NN	O	B-protein
beta	NN	O	I-protein
antibody	NN	O	I-protein
had	NN	O	O
negligible	NN	O	O
effects	NN	O	O
on	NN	O	O
HUVEC	NN	O	B-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
PAI-1	NN	O	B-protein
or	NN	O	O
vWf	NN	O	B-protein
production	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
promonocytic	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
require	NN	O	O
prestimulation	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
activate	NN	O	O
HUVEC	NN	O	B-cell_type
and	NN	O	O
that	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
contributes	NN	O	O
to	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
.	NN	O	O

-DOCSTART-	O

Engagement	NN	O	O
of	NN	O	O
the	NN	O	O
Lewis	NN	O	O
X	NN	O	O
antigen	NN	O	O
(	NN	O	O
CD15	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
monocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
tumor	NN	O	B-cell_line
necrosis	NN	O	I-cell_line
factor-alpha	NN	O	I-cell_line
(	NN	O	I-cell_line
TNF-alpha	NN	O	I-cell_line
)	NN	O	I-cell_line
-treated	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
tissue	NN	O	O
factor	NN	O	O
and	NN	O	O
CD36	NN	O	B-protein
on	NN	O	O
monocytes	NN	O	O
.	NN	O	O

Using	NN	O	O
immunological	NN	O	O
cross-linking	NN	O	O
to	NN	O	O
mimic	NN	O	O
receptor	NN	O	O
engagement	NN	O	O
by	NN	O	O
natural	NN	O	O
ligands	NN	O	O
,	NN	O	O
we	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
CD15	NN	O	O
(	NN	O	O
Lewis	NN	O	O
X	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
monocyte	NN	O	B-protein
counter-receptor	NN	O	I-protein
for	NN	O	O
endothelial	NN	O	B-protein
selectins	NN	O	I-protein
may	NN	O	O
participate	NN	O	O
in	NN	O	O
this	NN	O	O
response	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
cytokine	NN	O	O
production	NN	O	O
as	NN	O	O
a	NN	O	O
readout	NN	O	O
for	NN	O	O
monocyte	NN	O	O
activation	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
CD15	NN	O	O
cross-linking	NN	O	O
induced	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
from	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
cells	NN	O	O
from	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
MM6	NN	O	I-cell_line
.	NN	O	O

Quantitative	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
showed	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
steady-state	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
after	NN	O	O
3	NN	O	O
to	NN	O	O
4	NN	O	O
hours	NN	O	O
of	NN	O	O
cross-linking	NN	O	O
.	NN	O	O

CD15	NN	O	O
cross-linking	NN	O	O
also	NN	O	O
concomitantly	NN	O	O
increased	NN	O	O
interleukin-1	NN	O	B-RNA
beta	NN	O	I-RNA
(	NN	O	I-RNA
IL-1	NN	O	I-RNA
beta	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
while	NN	O	O
no	NN	O	O
apparent	NN	O	O
change	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
beta-actin	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
indicating	NN	O	O
specificity	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
by	NN	O	O
CD15	NN	O	O
engagement	NN	O	O
,	NN	O	O
a	NN	O	O
CAT	NN	O	O
plasmid	NN	O	O
reporter	NN	O	O
construct	NN	O	O
containing	NN	O	O
IL-1	NN	O	B-DNA
beta	NN	O	I-DNA
promoter/enhancer	NN	O	I-DNA
sequences	NN	O	I-DNA
was	NN	O	O
introduced	NN	O	O
into	NN	O	O
MM6	NN	O	B-cell_line
.	NN	O	O

Subsequent	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
CD15	NN	O	O
increased	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

CD15	NN	O	O
engagement	NN	O	O
by	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
also	NN	O	O
attenuated	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
transcript	NN	O	O
degradation	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
signaling	NN	O	O
via	NN	O	O
CD15	NN	O	O
also	NN	O	O
had	NN	O	O
posttranscriptional	NN	O	O
effects	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
anti-	NN	O	O
CD15	NN	O	B-cell_type
cross-linked	NN	O	I-cell_type
cells	NN	O	I-cell_type
demonstrated	NN	O	O
enhanced	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
factor	NN	O	I-protein
activator	NN	O	I-protein
protein-1	NN	O	I-protein
,	NN	O	O
minimally	NN	O	O
changed	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
SV40	NN	O	B-protein
promoter	NN	O	I-protein
specific	NN	O	I-protein
protein-1	NN	O	I-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
engagement	NN	O	O
of	NN	O	O
CD15	NN	O	O
on	NN	O	O
monocytes	NN	O	O
results	NN	O	O
in	NN	O	O
monocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
its	NN	O	O
well-recognized	NN	O	O
adhesive	NN	O	O
role	NN	O	O
,	NN	O	O
CD15	NN	O	O
may	NN	O	O
function	NN	O	O
as	NN	O	O
an	NN	O	O
important	NN	O	O
signaling	NN	O	O
molecule	NN	O	O
capable	NN	O	O
of	NN	O	O
initiating	NN	O	O
proinflammatory	NN	O	O
events	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
that	NN	O	O
come	NN	O	O
into	NN	O	O
contact	NN	O	O
with	NN	O	O
activated	NN	O	O
endothelium	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD2	NN	O	B-protein
signalling	NN	O	O
induces	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Promoter	NN	O	O
sequences	NN	O	O
responsive	NN	O	O
to	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
(	NN	O	O
cAMP	NN	O	O
)	NN	O	O
are	NN	O	O
found	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
cellular	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
bind	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
the	NN	O	O
cAMP	NN	O	B-protein
response	NN	O	I-protein
element	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	I-protein
/activating	NN	O	I-protein
transcription	NN	O	I-protein
factor-1	NN	O	I-protein
(	NN	O	I-protein
ATF-1	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
a	NN	O	O
human	NN	O	O
T-lymphotropic	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
model	NN	O	O
of	NN	O	O
cAMP	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CRE	NN	O	B-DNA
)	NN	O	O
transcription	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
on	NN	O	O
transcription	NN	O	O
from	NN	O	O
homologous	NN	O	O
regions	NN	O	O
in	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
increased	NN	O	O
HTLV-1	NN	O	O
transcription	NN	O	O
following	NN	O	O
CD2	NN	O	O
but	NN	O	O
not	NN	O	O
CD3	NN	O	O
receptor	NN	O	O
cross-linking	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
this	NN	O	O
increased	NN	O	O
viral	NN	O	O
transcription	NN	O	O
was	NN	O	O
mediated	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
through	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
CD2	NN	O	B-protein
and	NN	O	O
CD3	NN	O	B-protein
receptor	NN	O	I-protein
-mediated	NN	O	O
signalling	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

CD2	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
CD3	NN	O	B-protein
,	NN	O	O
cross-linking	NN	O	O
increased	NN	O	O
cAMP	NN	O	O
detected	NN	O	O
by	NN	O	O
competitive	NN	O	O
enzyme-linked	NN	O	O
immunosorbent	NN	O	O
assay	NN	O	O
(	NN	O	O
ELISA	NN	O	O
)	NN	O	O
approximately	NN	O	O
fourfold	NN	O	O
.	NN	O	O

CD2	NN	O	B-protein
cross-linking	NN	O	O
concurrently	NN	O	O
increased	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
detected	NN	O	O
by	NN	O	O
immunoblot	NN	O	O
assay	NN	O	O
eightfold	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
post-translational	NN	O	O
regulation	NN	O	O
,	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
total	NN	O	O
level	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
protein	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
occurred	NN	O	O
through	NN	O	O
a	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
and	NN	O	O
Rp-cAMP-	NN	O	O
sensitive	NN	O	O
pathway	NN	O	O
,	NN	O	O
suggesting	NN	O	O
phosphorylation	NN	O	O
required	NN	O	O
antecedent	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
(	NN	O	O
PTK	NN	O	B-protein
)	NN	O	O
and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
(	NN	O	O
PKA	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Both	NN	O	O
CD2	NN	O	B-protein
and	NN	O	O
CD3	NN	O	B-protein
cross-linking	NN	O	O
increased	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
a	NN	O	O
radiolabelled	NN	O	O
CRE	NN	O	O
oligonucleotide	NN	O	O
probe	NN	O	O
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
suggesting	NN	O	O
that	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
enhances	NN	O	O
binding	NN	O	O
independently	NN	O	O
of	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
at	NN	O	O
serine	NN	O	O
133	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
specific	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
CREB/ATF-1	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
by	NN	O	O
the	NN	O	O
CD2	NN	O	B-protein
signalling	NN	O	O
pathway	NN	O	O
and	NN	O	O
suggest	NN	O	O
CD2	NN	O	B-protein
receptor	NN	O	O
modulation	NN	O	O
of	NN	O	O
CRE	NN	O	B-DNA
-mediated	NN	O	O
transcription	NN	O	O
following	NN	O	O
ligand	NN	O	O
engagement	NN	O	O
(	NN	O	O
e.g.	NN	O	O
cell-to-cell	NN	O	O
contact	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

X-rays-induced	NN	O	O
secretion	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
that	NN	O	O
enhance	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
HIV-1	NN	O	B-DNA
promoter	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
various	NN	O	O
non-irradiated	NN	O	B-cell_line
transfected	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Various	NN	O	O
cellular	NN	O	O
stress	NN	O	O
agents	NN	O	O
like	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
exposure	NN	O	O
could	NN	O	O
activate	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-	NN	O	O
1	NN	O	O
)	NN	O	O
replication	NN	O	O
or	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
extracellular	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
released	NN	O	O
by	NN	O	O
X-ray-treated	NN	O	B-cell_line
human	NN	O	I-cell_line
colonic	NN	O	I-cell_line
carcinoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
HT29	NN	O	B-cell_line
)	NN	O	O
might	NN	O	O
activate	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
non-irradiated	NN	O	B-cell_line
HT29	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
report	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
various	NN	O	O
transiently	NN	O	O
or	NN	O	O
stably	NN	O	B-cell_line
transfected	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
X-ray	NN	O	O
irradiation	NN	O	O
up-regulates	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
transcription	NN	O	O
through	NN	O	O
the	NN	O	O
kappaB	NN	O	B-protein
regulatory	NN	O	I-protein
elements	NN	O	I-protein
.	NN	O	O

A	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
processed	NN	O	O
by	NN	O	O
and	NN	O	O
acts	NN	O	O
upon	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
was	NN	O	O
detected	NN	O	O
by	NN	O	O
addition	NN	O	O
to	NN	O	O
non-irradiated	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
either	NN	O	O
X-ray-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
a	NN	O	O
conditioned	NN	O	O
medium	NN	O	O
taken	NN	O	O
from	NN	O	O
irradiated	NN	O	B-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
magnitude	NN	O	O
of	NN	O	O
responsiveness	NN	O	O
is	NN	O	O
cell	NN	O	O
type	NN	O	O
dependent	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
X-ray	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
in	NN	O	O
transiently	NN	O	O
or	NN	O	O
stably	NN	O	O
transfected	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
a	NN	O	O
potent	NN	O	O
antioxidant	NN	O	O
drug	NN	O	O
,	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
and	NN	O	O
by	NN	O	O
another	NN	O	O
drug	NN	O	O
,	NN	O	O
known	NN	O	O
for	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
trapping	NN	O	O
of	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
suramin	NN	O	O
.	NN	O	O

The	NN	O	O
importance	NN	O	O
of	NN	O	O
these	NN	O	O
observations	NN	O	O
in	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
AIDS-related	NN	O	O
cancers	NN	O	O
treated	NN	O	O
by	NN	O	O
radiotherapy	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
established	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
delta-globin	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
primary	NN	O	B-cell_type
adult	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Restoration	NN	O	O
of	NN	O	O
the	NN	O	O
CCAAT	NN	O	B-DNA
box	NN	O	I-DNA
or	NN	O	O
insertion	NN	O	O
of	NN	O	O
an	NN	O	O
erythroid	NN	O	B-DNA
Kruppel-like	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
EKLF	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
delta	NN	O	B-DNA
promoter	NN	O	I-DNA
activates	NN	O	O
its	NN	O	O
expression	NN	O	O
in	NN	O	O
several	NN	O	O
erythroid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
extended	NN	O	O
these	NN	O	O
studies	NN	O	O
using	NN	O	O
a	NN	O	O
novel	NN	O	O
primary	NN	O	B-cell_line
human	NN	O	I-cell_line
adult	NN	O	I-cell_line
erythroid	NN	O	I-cell_line
cell	NN	O	I-cell_line
(	NN	O	I-cell_line
hAEC	NN	O	I-cell_line
)	NN	O	I-cell_line
system	NN	O	I-cell_line
to	NN	O	O
investigate	NN	O	O
these	NN	O	O
effects	NN	O	O
at	NN	O	O
the	NN	O	O
late	NN	O	B-cell_type
erythroblast	NN	O	I-cell_type
stage	NN	O	I-cell_type
.	NN	O	O

Restoration	NN	O	O
of	NN	O	O
the	NN	O	O
CCAAT	NN	O	B-DNA
box	NN	O	I-DNA
at	NN	O	O
-70	NN	O	B-DNA
bp	NN	O	I-DNA
,	NN	O	O
or	NN	O	O
insertion	NN	O	O
of	NN	O	O
an	NN	O	O
EKLF	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
at	NN	O	O
-85	NN	O	B-DNA
bp	NN	O	I-DNA
or	NN	O	O
-95	NN	O	B-DNA
bp	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
significantly	NN	O	O
increased	NN	O	O
delta	NN	O	O
globin	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
hAEC	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
altered	NN	O	O
CCAAT	NN	O	B-DNA
box	NN	O	I-DNA
(	NN	O	O
CCAAC	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
an	NN	O	O
EKLF	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
delta-globin	NN	O	B-protein
contribute	NN	O	O
to	NN	O	O
its	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
hAEC	NN	O	B-cell_line
model	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
association	NN	O	O
of	NN	O	O
JAK1	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
in	NN	O	O
pro-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
dissolved	NN	O	O
by	NN	O	O
interleukin-4	NN	O	B-protein
-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
both	NN	O	O
proteins	NN	O	O
.	NN	O	O

The	NN	O	O
bipartite	NN	O	B-protein
human	NN	O	I-protein
interleukin-4	NN	O	I-protein
(	NN	O	I-protein
IL-4	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
was	NN	O	O
functionally	NN	O	O
expressed	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
pro-B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
activated	NN	O	O
by	NN	O	O
human	NN	O	B-protein
IL-4	NN	O	I-protein
to	NN	O	O
evoke	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
.	NN	O	O

Mutual	NN	O	O
association	NN	O	O
of	NN	O	O
signal	NN	O	B-protein
transducing	NN	O	I-protein
proteins	NN	O	I-protein
within	NN	O	O
the	NN	O	O
receptor	NN	O	O
complex	NN	O	O
was	NN	O	O
then	NN	O	O
studied	NN	O	O
in	NN	O	O
dependence	NN	O	O
of	NN	O	O
ligand	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Besides	NN	O	O
ligand-induced	NN	O	O
receptor	NN	O	O
heterodimerization	NN	O	O
and	NN	O	O
contacts	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
IL-4	NN	O	B-protein
receptor	NN	O	I-protein
subunits	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
gamma	NN	O	I-protein
with	NN	O	O
Janus	NN	O	B-protein
kinases	NN	O	I-protein
JAK1	NN	O	B-protein
and	NN	O	O
JAK3	NN	O	B-protein
a	NN	O	O
prominent	NN	O	O
constitutive	NN	O	O
binding	NN	O	O
between	NN	O	O
JAK1	NN	O	B-protein
and	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
STAT5	NN	O	B-protein
was	NN	O	O
detected	NN	O	O
.	NN	O	O

Since	NN	O	O
both	NN	O	O
these	NN	O	O
proteins	NN	O	O
become	NN	O	O
phosphorylated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
receptor	NN	O	I-protein
stimulation	NN	O	O
,	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
on	NN	O	O
their	NN	O	O
mutual	NN	O	O
contact	NN	O	O
was	NN	O	O
analyzed	NN	O	O
.	NN	O	O

Association	NN	O	O
of	NN	O	O
JAK1	NN	O	B-protein
and	NN	O	O
STAT5	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
occur	NN	O	O
exclusively	NN	O	O
between	NN	O	O
unphosphorylated	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
by	NN	O	O
mechanical	NN	O	O
ventilation	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Positive-pressure	NN	O	O
mechanical	NN	O	O
ventilation	NN	O	O
supports	NN	O	O
gas	NN	O	O
exchange	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
respiratory	NN	O	O
failure	NN	O	O
but	NN	O	O
is	NN	O	O
also	NN	O	O
responsible	NN	O	O
for	NN	O	O
significant	NN	O	O
lung	NN	O	O
injury	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
developed	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
isolated	NN	O	O
lung	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
be	NN	O	O
submitted	NN	O	O
to	NN	O	O
a	NN	O	O
prolonged	NN	O	B-cell_line
cyclic	NN	O	I-cell_line
pressure-stretching	NN	O	I-cell_line
strain	NN	O	I-cell_line
resembling	NN	O	O
that	NN	O	O
of	NN	O	O
conventional	NN	O	O
mechanical	NN	O	O
ventilation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
model	NN	O	O
,	NN	O	O
cells	NN	O	O
cultured	NN	O	O
on	NN	O	O
a	NN	O	O
Silastic	NN	O	O
membrane	NN	O	O
were	NN	O	O
elongated	NN	O	O
up	NN	O	O
to	NN	O	O
7	NN	O	O
%	NN	O	O
of	NN	O	O
their	NN	O	O
initial	NN	O	O
diameter	NN	O	O
,	NN	O	O
corresponding	NN	O	O
to	NN	O	O
a	NN	O	O
12	NN	O	O
%	NN	O	O
increase	NN	O	O
in	NN	O	O
cell	NN	O	O
surface	NN	O	O
.	NN	O	O

The	NN	O	O
lung	NN	O	B-cell_type
macrophage	NN	O	I-cell_type
was	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
main	NN	O	O
cellular	NN	O	O
source	NN	O	O
for	NN	O	O
critical	NN	O	O
inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
such	NN	O	O
as	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
,	NN	O	O
the	NN	O	O
chemokines	NN	O	B-protein
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-8	NN	O	I-protein
and	NN	O	I-protein
-6	NN	O	I-protein
,	NN	O	O
and	NN	O	O
matrix	NN	O	B-protein
metalloproteinase-9	NN	O	I-protein
in	NN	O	O
this	NN	O	O
model	NN	O	O
system	NN	O	O
of	NN	O	O
mechanical	NN	O	O
ventilation	NN	O	O
.	NN	O	O

These	NN	O	O
mediators	NN	O	O
were	NN	O	O
measured	NN	O	O
in	NN	O	O
supernatants	NN	O	O
from	NN	O	O
ventilated	NN	O	B-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
promonocytic	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
activated	NN	O	O
in	NN	O	O
ventilated	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Synergistic	NN	O	O
proinflammatory	NN	O	O
effects	NN	O	O
of	NN	O	O
mechanical	NN	O	O
stress	NN	O	O
and	NN	O	O
molecules	NN	O	O
such	NN	O	O
as	NN	O	O
bacterial	NN	O	B-protein
endotoxin	NN	O	I-protein
were	NN	O	O
observed	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
mechanical	NN	O	O
ventilation	NN	O	O
might	NN	O	O
be	NN	O	O
particularly	NN	O	O
deleterious	NN	O	O
in	NN	O	O
preinjured	NN	O	O
or	NN	O	O
infected	NN	O	O
lungs	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
prevented	NN	O	O
IL-8	NN	O	B-protein
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
secretion	NN	O	O
in	NN	O	O
ventilated	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Mechanical	NN	O	O
ventilation	NN	O	O
induced	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-8	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
alveolar	NN	O	B-cell_type
type	NN	O	I-cell_type
II-like	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Other	NN	O	O
lung	NN	O	O
cell	NN	O	O
types	NN	O	O
such	NN	O	O
as	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
bronchial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
fibroblasts	NN	O	B-cell_type
failed	NN	O	O
to	NN	O	O
produce	NN	O	O
IL-8	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
prolonged	NN	O	O
cyclic	NN	O	O
pressure-stretching	NN	O	O
load	NN	O	O
.	NN	O	O

This	NN	O	O
model	NN	O	O
is	NN	O	O
of	NN	O	O
particular	NN	O	O
value	NN	O	O
for	NN	O	O
exploring	NN	O	O
physical	NN	O	O
stress-induced	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
for	NN	O	O
testing	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
novel	NN	O	O
ventilatory	NN	O	O
strategies	NN	O	O
or	NN	O	O
adjunctive	NN	O	O
substances	NN	O	O
aimed	NN	O	O
at	NN	O	O
modulating	NN	O	O
cell	NN	O	O
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
mechanical	NN	O	O
ventilation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signaling	NN	O	O
pathways	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
TNF-	NN	O	B-protein
and	NN	O	I-protein
cytokine-receptor	NN	O	I-protein
families	NN	O	O
target	NN	O	O
a	NN	O	O
common	NN	O	O
cis-element	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
IFN	NN	O	B-DNA
regulatory	NN	O	I-DNA
factor	NN	O	I-DNA
1	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

CD40	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
strongly	NN	O	O
influenced	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
interplay	NN	O	O
between	NN	O	O
these	NN	O	O
distinct	NN	O	O
stimuli	NN	O	O
is	NN	O	O
not	NN	O	O
clearly	NN	O	O
delineated	NN	O	O
.	NN	O	O

IFN	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
IRF-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
activated	NN	O	O
by	NN	O	O
either	NN	O	O
CD40	NN	O	B-protein
or	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
these	NN	O	O
different	NN	O	O
sets	NN	O	O
of	NN	O	O
signals	NN	O	O
target	NN	O	O
a	NN	O	O
common	NN	O	O
cis-acting	NN	O	O
element	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
,	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-DNA
gamma-activated	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
GAS	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Targeting	NN	O	O
of	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-DNA
GAS	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
confined	NN	O	O
to	NN	O	O
activation	NN	O	O
via	NN	O	O
CD40	NN	O	B-protein
but	NN	O	O
extends	NN	O	O
to	NN	O	O
other	NN	O	O
stimuli	NN	O	O
that	NN	O	O
mimic	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
cascade	NN	O	O
,	NN	O	O
like	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
EBV	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
induction	NN	O	O
of	NN	O	O
STATs	NN	O	B-protein
by	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-DNA
GAS	NN	O	I-DNA
-binding	NN	O	O
complex	NN	O	O
activated	NN	O	O
by	NN	O	O
CD40	NN	O	B-protein
,	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
or	NN	O	O
EBV	NN	O	O
contains	NN	O	O
Rel	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
specifically	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
system	NN	O	O
,	NN	O	O
simultaneous	NN	O	O
exposure	NN	O	O
to	NN	O	O
CD40L	NN	O	B-protein
together	NN	O	O
with	NN	O	O
either	NN	O	O
IL-4	NN	O	B-protein
or	NN	O	O
IFN-gamma	NN	O	B-protein
does	NN	O	O
not	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
novel	NN	O	O
Rel/STAT	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

Given	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
IRF-1	NN	O	B-protein
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
biologic	NN	O	O
functions	NN	O	O
from	NN	O	O
proliferation	NN	O	O
to	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
support	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
modulation	NN	O	O
of	NN	O	O
IRF-1	NN	O	B-protein
levels	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
critical	NN	O	O
control	NN	O	O
point	NN	O	O
in	NN	O	O
B	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
effects	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
inhibitors	NN	O	O
on	NN	O	O
fibronectin	NN	O	B-protein
-induced	NN	O	O
interleukin-beta	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
,	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
and	NN	O	O
secretion	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
interleukin-1beta	NN	O	B-protein
(	NN	O	O
IL-1beta	NN	O	B-protein
)	NN	O	O
when	NN	O	O
stimulated	NN	O	O
with	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
matrix	NN	O	I-protein
glycoprotein	NN	O	I-protein
,	NN	O	O
fibronectin	NN	O	B-protein
(	NN	O	O
FN	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
is	NN	O	O
considered	NN	O	O
important	NN	O	O
for	NN	O	O
this	NN	O	O
process	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
metabolic	NN	O	O
steps	NN	O	O
at	NN	O	O
which	NN	O	O
PKC	NN	O	B-protein
acts	NN	O	O
upon	NN	O	O
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
FN	NN	O	B-protein
-induced	NN	O	O
IL-1beta	NN	O	B-protein
response	NN	O	O
remain	NN	O	O
unclear	NN	O	O
.	NN	O	O

We	NN	O	O
performed	NN	O	O
an	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
two	NN	O	O
PKC	NN	O	B-protein
inhibitors	NN	O	O
,	NN	O	O
Calphostin	NN	O	O
C	NN	O	O
and	NN	O	O
Staurosporine	NN	O	O
,	NN	O	O
prevent	NN	O	O
the	NN	O	O
FN	NN	O	B-protein
-induced	NN	O	O
IL-1beta	NN	O	B-protein
response	NN	O	O
.	NN	O	O

Both	NN	O	O
inhibitors	NN	O	O
blocked	NN	O	O
the	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
protein	NN	O	I-protein
into	NN	O	O
the	NN	O	O
media	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
exposed	NN	O	O
to	NN	O	O
FN	NN	O	B-protein
.	NN	O	O

Immunoprecipitation	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
unde3r	NN	O	O
these	NN	O	O
circumstances	NN	O	O
,	NN	O	O
Calphostin	NN	O	O
C	NN	O	O
inhibited	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
Staurosporine	NN	O	O
allowed	NN	O	O
protein	NN	O	O
production	NN	O	O
,	NN	O	O
but	NN	O	O
inhibited	NN	O	O
its	NN	O	O
secretion	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
responsible	NN	O	O
for	NN	O	O
these	NN	O	O
differences	NN	O	O
,	NN	O	O
we	NN	O	O
turned	NN	O	O
to	NN	O	O
human	NN	O	B-cell_line
U937	NN	O	I-cell_line
promonocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

U937	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	O
full-length	NN	O	B-DNA
IL-1beta	NN	O	I-DNA
promoter	NN	O	I-DNA
connected	NN	O	O
to	NN	O	O
a	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
were	NN	O	O
submitted	NN	O	O
to	NN	O	O
transcription	NN	O	O
assays	NN	O	O
,	NN	O	O
Northern	NN	O	O
blotting	NN	O	O
,	NN	O	O
and	NN	O	O
DNA	NN	O	O
electrophoresis	NN	O	O
mobility	NN	O	O
gel	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
revealed	NN	O	O
that	NN	O	O
Calphostin	NN	O	O
C	NN	O	O
inhibited	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
activator	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
which	NN	O	O
is	NN	O	O
considered	NN	O	O
necessary	NN	O	O
for	NN	O	O
FN	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
prevented	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1beta	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
Staurosporine	NN	O	O
alone	NN	O	O
induced	NN	O	O
AP-1	NN	O	B-protein
translocation	NN	O	O
and	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
.	NN	O	O

Overall	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
Calphostin	NN	O	O
C	NN	O	O
prevents	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1beta	NN	O	B-DNA
gene	NN	O	I-DNA
thereby	NN	O	O
inhibiting	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Based	NN	O	O
on	NN	O	O
the	NN	O	O
high	NN	O	O
specificity	NN	O	O
of	NN	O	O
this	NN	O	O
compound	NN	O	O
for	NN	O	O
PKC	NN	O	B-protein
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
PKC	NN	O	B-protein
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
FN	NN	O	B-protein
-induced	NN	O	O
IL-1beta	NN	O	B-protein
protein	NN	O	O
production	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
Staurosporine	NN	O	O
prevented	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
by	NN	O	O
unknown	NN	O	O
mechanisms	NN	O	O

-DOCSTART-	O

The	NN	O	O
carboxyl-terminal	NN	O	B-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
CD36	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
oxidized	NN	O	O
low-density	NN	O	O
lipoprotein	NN	O	O
modulation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
oxidized	NN	O	O
low-density	NN	O	O
lipoprotein	NN	O	O
(	NN	O	O
Ox	NN	O	O
LDL	NN	O	O
)	NN	O	O
by	NN	O	O
monocyte	NN	O	B-cell_type
-macrophages	NN	O	B-cell_type
causes	NN	O	O
pleiotropic	NN	O	O
effects	NN	O	O
,	NN	O	O
including	NN	O	O
changes	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
represent	NN	O	O
an	NN	O	O
early	NN	O	O
event	NN	O	O
in	NN	O	O
atherogenesis	NN	O	O
.	NN	O	O

The	NN	O	O
integral	NN	O	O
membrane	NN	O	O
glycoprotein	NN	O	B-protein
CD36	NN	O	I-protein
appears	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
physiological	NN	O	O
role	NN	O	O
in	NN	O	O
binding	NN	O	O
and	NN	O	O
uptake	NN	O	O
of	NN	O	O
Ox	NN	O	O
LDL	NN	O	O
by	NN	O	O
monocyte-macrophages	NN	O	B-cell_type
,	NN	O	O
although	NN	O	O
the	NN	O	O
molecular	NN	O	O
events	NN	O	O
associated	NN	O	O
with	NN	O	O
CD36	NN	O	B-protein
-Ox	NN	O	O
LDL	NN	O	O
interaction	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

To	NN	O	O
approach	NN	O	O
this	NN	O	O
issue	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
CD36	NN	O	B-protein
transfected	NN	O	O
Chinese	NN	O	B-cell_type
hampster	NN	O	I-cell_type
ovary	NN	O	I-cell_type
(	NN	O	I-cell_type
CHO	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
exposed	NN	O	O
them	NN	O	O
to	NN	O	O
Ox	NN	O	O
LDL	NN	O	O
,	NN	O	O
and	NN	O	O
determined	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
Ox	NN	O	O
LDL	NN	O	O
enhanced	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
to	NN	O	O
an	NN	O	O
NF-kappaB	NN	O	B-DNA
sequence	NN	O	I-DNA
following	NN	O	O
activation	NN	O	O
of	NN	O	O
CD36-producing	NN	O	B-cell_line
CHO	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
proinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

This	NN	O	O
enhanced	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
coincubation	NN	O	O
of	NN	O	O
CD36	NN	O	B-protein
transfected	NN	O	O
cells	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	B-protein
CD36-specific	NN	O	I-protein
antibody	NN	O	I-protein
OKM5	NN	O	B-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
determined	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
required	NN	O	O
an	NN	O	O
intact	NN	O	O
carboxyl-terminal	NN	O	B-protein
cytoplasmic	NN	O	I-protein
segment	NN	O	I-protein
on	NN	O	O
CD36	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
idea	NN	O	O
that	NN	O	O
human	NN	O	B-protein
CD36	NN	O	I-protein
mediates	NN	O	O
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
Ox	NN	O	O
LDL	NN	O	O
.	NN	O	O

-DOCSTART-	O

TRAMP	NN	O	B-protein
,	NN	O	O
a	NN	O	O
novel	NN	O	O
apoptosis-mediating	NN	O	B-protein
receptor	NN	O	I-protein
with	NN	O	O
sequence	NN	O	O
homology	NN	O	O
to	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
Fas	NN	O	B-protein
(	NN	O	O
Apo-1/CD95	NN	O	B-protein
)	NN	O	O
.	NN	O	O

A	NN	O	O
novel	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
designated	NN	O	O
TRAMP	NN	O	B-protein
,	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

The	NN	O	O
structural	NN	O	O
organization	NN	O	O
of	NN	O	O
the	NN	O	O
393	NN	O	B-protein
amino	NN	O	I-protein
acid	NN	O	I-protein
long	NN	O	I-protein
human	NN	O	I-protein
TRAMP	NN	O	I-protein
is	NN	O	O
most	NN	O	O
homologous	NN	O	O
to	NN	O	O
TNF	NN	O	B-protein
receptor	NN	O	I-protein
1	NN	O	I-protein
.	NN	O	O

TRAMP	NN	O	B-protein
is	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
on	NN	O	O
thymocytes	NN	O	B-cell_type
and	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Its	NN	O	O
extracellular	NN	O	O
domain	NN	O	O
is	NN	O	O
composed	NN	O	O
of	NN	O	O
four	NN	O	O
cysteine-rich	NN	O	B-protein
domains	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
region	NN	O	I-protein
contains	NN	O	O
a	NN	O	O
death	NN	O	B-protein
domain	NN	O	I-protein
known	NN	O	O
to	NN	O	O
signal	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
TRAMP	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
two	NN	O	O
major	NN	O	O
responses	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

TRAMP	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
death	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
ICE-like	NN	O	B-protein
proteases	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
Bcl-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
TRAMP	NN	O	B-protein
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
known	NN	O	O
apoptosis-inducing	NN	O	B-protein
ligands	NN	O	I-protein
of	NN	O	O
the	NN	O	O
TNF	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
state	NN	O	O
of	NN	O	O
maturation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
into	NN	O	O
macrophages	NN	O	B-cell_type
determines	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
on	NN	O	O
HIV	NN	O	O
replication	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
on	NN	O	O
HIV	NN	O	O
infection	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
as	NN	O	O
they	NN	O	O
matured	NN	O	O
into	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
over	NN	O	O
7	NN	O	O
days	NN	O	O
were	NN	O	O
investigated	NN	O	O
using	NN	O	O
HIV-1	NN	O	O
(	NN	O	O
BaL	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
low	NN	O	O
passage	NN	O	O
clinical	NN	O	O
strains	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
up-regulated	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
genomic	NN	O	O
and	NN	O	O
spliced	NN	O	O
HIV	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
monocytes	NN	O	B-cell_type
cultured	NN	O	O
on	NN	O	O
Teflon	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
Northern	NN	O	O
analysis	NN	O	O
and	NN	O	O
p24	NN	O	B-protein
Ag	NN	O	I-protein
assay	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
nuclear	NN	O	O
run-on	NN	O	O
assay	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
stimulation	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
enhance	NN	O	O
transcription	NN	O	O
by	NN	O	O
two-	NN	O	O
to	NN	O	O
threefold	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
stimulated	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
binding	NN	O	O
before	NN	O	O
enhancement	NN	O	O
of	NN	O	O
HIV	NN	O	B-RNA
RNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
markedly	NN	O	O
and	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
HIV	NN	O	O
replication	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
in	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
this	NN	O	O
occurred	NN	O	O
whether	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
were	NN	O	O
added	NN	O	O
before	NN	O	O
or	NN	O	O
after	NN	O	O
HIV	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
reversal	NN	O	O
from	NN	O	O
stimulation	NN	O	O
to	NN	O	O
inhibition	NN	O	O
occurred	NN	O	O
after	NN	O	O
3	NN	O	O
to	NN	O	O
5	NN	O	O
days	NN	O	O
of	NN	O	O
adherence	NN	O	O
to	NN	O	O
plastic	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
had	NN	O	O
no	NN	O	O
significant	NN	O	O
effect	NN	O	O
on	NN	O	O
HIV	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
both	NN	O	O
cytokines	NN	O	B-protein
on	NN	O	O
the	NN	O	O
monocyte	NN	O	O
maturation/differentiation	NN	O	O
(	NN	O	O
CD11b	NN	O	B-protein
,	NN	O	O
CD13	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CD26	NN	O	B-protein
)	NN	O	O
and	NN	O	O
other	NN	O	O
macrophage	NN	O	B-protein
markers	NN	O	I-protein
(	NN	O	O
CD14	NN	O	B-protein
and	NN	O	O
CD68	NN	O	B-protein
)	NN	O	O
was	NN	O	O
examined	NN	O	O
.	NN	O	O

IL-4	NN	O	B-protein
enhanced	NN	O	O
CD11b	NN	O	B-protein
,	NN	O	O
but	NN	O	O
inhibited	NN	O	O
CD26	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
delayed	NN	O	O
CD13	NN	O	B-protein
loss	NN	O	O
.	NN	O	O

IL-13	NN	O	B-protein
had	NN	O	O
similar	NN	O	O
effects	NN	O	O
on	NN	O	O
CD11b	NN	O	B-protein
and	NN	O	O
CD13	NN	O	B-protein
,	NN	O	O
but	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
CD26	NN	O	B-protein
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
do	NN	O	O
not	NN	O	O
simply	NN	O	O
enhance	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
,	NN	O	O
but	NN	O	O
have	NN	O	O
complex	NN	O	O
and	NN	O	O
slightly	NN	O	O
divergent	NN	O	O
effects	NN	O	O
that	NN	O	O
impact	NN	O	O
on	NN	O	O
HIV	NN	O	O
replication	NN	O	O
probably	NN	O	O
through	NN	O	O
cell	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
translocation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mice	NN	O	O
lacking	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
CIITA	NN	O	B-protein
--	NN	O	O
a	NN	O	O
second	NN	O	O
look	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
generated	NN	O	O
a	NN	O	O
second	NN	O	O
line	NN	O	O
of	NN	O	O
mice	NN	O	O
lacking	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
thought	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
critical	NN	O	O
regulator	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
CIITA	NN	O	B-protein
(	NN	O	O
for	NN	O	O
class	NN	O	O
II	NN	O	O
transactivator	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Our	NN	O	O
and	NN	O	O
the	NN	O	O
previously	NN	O	O
published	NN	O	O
lines	NN	O	O
differ	NN	O	O
in	NN	O	O
the	NN	O	O
deletion	NN	O	O
that	NN	O	O
was	NN	O	O
engineered	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
we	NN	O	O
removed	NN	O	O
the	NN	O	O
neomycin-resistance	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
structural	NN	O	B-DNA
gene	NN	O	I-DNA
via	NN	O	O
the	NN	O	O
cre-loxP	NN	O	O
recombination	NN	O	O
system	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
our	NN	O	O
line	NN	O	O
led	NN	O	O
to	NN	O	O
two	NN	O	O
new	NN	O	O
findings	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
a	NN	O	O
substantial	NN	O	O
number	NN	O	O
of	NN	O	O
cells	NN	O	O
can	NN	O	O
express	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
,	NN	O	O
albeit	NN	O	O
at	NN	O	O
5-fold	NN	O	O
reduced	NN	O	O
levels	NN	O	O
,	NN	O	O
most	NN	O	O
notably	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
s.c.	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
;	NN	O	O
therefore	NN	O	O
,	NN	O	O
the	NN	O	O
CIITA	NN	O	B-DNA
gene	NN	O	I-DNA
can	NN	O	O
not	NN	O	O
be	NN	O	O
an	NN	O	O
absolute	NN	O	O
'	NN	O	O
master	NN	O	B-DNA
gene	NN	O	I-DNA
'	NN	O	O
controlling	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
,	NN	O	O
as	NN	O	O
had	NN	O	O
been	NN	O	O
thought	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
recent	NN	O	O
results	NN	O	O
on	NN	O	O
human	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
CIITA	NN	O	B-protein
is	NN	O	O
not	NN	O	O
critically	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
I	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Interleukin-10	NN	O	B-protein
stabilizes	NN	O	O
inhibitory	NN	O	O
kappaB-alpha	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Interleukin-10	NN	O	B-protein
(	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
protects	NN	O	O
animals	NN	O	O
from	NN	O	O
lethal	NN	O	O
endotoxemia	NN	O	O
.	NN	O	O

This	NN	O	O
beneficial	NN	O	O
effect	NN	O	O
is	NN	O	O
mediated	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
production	NN	O	O
,	NN	O	O
including	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Evidence	NN	O	O
suggests	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
may	NN	O	O
inhibit	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
through	NN	O	O
an	NN	O	O
unknown	NN	O	O
mechanism	NN	O	O
.	NN	O	O

NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
inflammatory	NN	O	O
signals	NN	O	O
is	NN	O	O
dependent	NN	O	O
upon	NN	O	O
degradation	NN	O	O
of	NN	O	O
its	NN	O	O
associated	NN	O	O
inhibitory	NN	O	O
peptide	NN	O	O
,	NN	O	O
inhibitory	NN	O	B-protein
kappaB-alpha	NN	O	I-protein
(	NN	O	O
IkappaB-alpha	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
IL-10	NN	O	B-protein
prevents	NN	O	O
human	NN	O	O
monocyte	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
resultant	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
by	NN	O	O
stabilization	NN	O	O
of	NN	O	O
IkappaB-alpha	NN	O	B-protein
.	NN	O	O

The	NN	O	O
purpose	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
IL-10	NN	O	B-protein
on	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-induced	NN	O	O
human	NN	O	O
monocyte	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
and	NN	O	O
IkappaB-alpha	NN	O	B-protein
degradation	NN	O	O
.	NN	O	O

Monocytes	NN	O	O
were	NN	O	O
isolated	NN	O	O
from	NN	O	O
human	NN	O	O
donors	NN	O	O
.	NN	O	O

Cells	NN	O	O
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
endotoxin	NN	O	B-protein
(	NN	O	O
LPS	NN	O	O
,	NN	O	O
100	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
with	NN	O	O
and	NN	O	O
without	NN	O	O
human	NN	O	O
IL-10	NN	O	B-protein
(	NN	O	O
10	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
.	NN	O	O

Following	NN	O	O
stimulation	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-protein
was	NN	O	O
measured	NN	O	O
in	NN	O	O
cell	NN	O	O
supernatants	NN	O	O
by	NN	O	O
ELISA	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
and	NN	O	O
IkappaB-alpha	NN	O	B-protein
levels	NN	O	O
by	NN	O	O
Western	NN	O	O
blot	NN	O	O
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
after	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
TNF-alpha	NN	O	B-protein
increased	NN	O	O
to	NN	O	O
798+/-67	NN	O	O
pg/mL	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
.001	NN	O	O
versus	NN	O	O
control	NN	O	O
)	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
attenuated	NN	O	O
LPS-stimulated	NN	O	O
TNF-alpha	NN	O	B-protein
production	NN	O	O
(	NN	O	O
297+/-54	NN	O	O
;	NN	O	O
p	NN	O	O
<	NN	O	O
.001	NN	O	O
versus	NN	O	O
LPS	NN	O	O
alone	NN	O	O
)	NN	O	O
.	NN	O	O

After	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
IkappaB-alpha	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
decreased	NN	O	O
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
increased	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
pretreatment	NN	O	O
prevented	NN	O	O
LPS-induced	NN	O	O
decreases	NN	O	O
in	NN	O	O
IkappaB-alpha	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
and	NN	O	O
attenuated	NN	O	O
NF-kappaB	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

IL-10	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
prevent	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
by	NN	O	O
preserving	NN	O	O
IkappaB-alpha	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
a	NN	O	O
reduction	NN	O	O
in	NN	O	O
TNF-alpha	NN	O	B-protein
release	NN	O	O
.	NN	O	O

-DOCSTART-	O

Down-regulation	NN	O	O
of	NN	O	O
human	NN	O	O
granzyme	NN	O	B-protein
B	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
inhibits	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
Ikaros	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
granzyme	NN	O	B-DNA
B	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
serine	NN	O	B-protein
protease	NN	O	I-protein
granzyme	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
an	NN	O	O
essential	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
granule	NN	O	O
exocytosis	NN	O	O
pathway	NN	O	O
,	NN	O	O
a	NN	O	O
major	NN	O	O
apoptotic	NN	O	O
mechanism	NN	O	O
used	NN	O	O
by	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
induce	NN	O	O
target	NN	O	O
cell	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Granzyme	NN	O	O
B	NN	O	O
gene	NN	O	O
transcription	NN	O	O
is	NN	O	O
induced	NN	O	O
in	NN	O	O
activated	NN	O	O
lymphocytes	NN	O	B-cell_type
upon	NN	O	O
antigenic	NN	O	O
stimulation	NN	O	O
,	NN	O	O
and	NN	O	O
several	NN	O	O
regulatory	NN	O	B-DNA
regions	NN	O	I-DNA
including	NN	O	O
CBF	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Ikaros	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
essential	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
granzyme	NN	O	B-DNA
B	NN	O	I-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
,	NN	O	O
a	NN	O	O
glucocorticoid	NN	O	O
that	NN	O	O
is	NN	O	O
widely	NN	O	O
used	NN	O	O
as	NN	O	O
an	NN	O	O
immunomodulatory	NN	O	O
and	NN	O	O
anti-inflammatory	NN	O	O
agent	NN	O	O
,	NN	O	O
inhibits	NN	O	O
granzyme	NN	O	B-RNA
B	NN	O	I-RNA
mRNA	NN	O	I-RNA
transcript	NN	O	I-RNA
in	NN	O	O
phytohemagglutinin-activated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	O
construct	NN	O	O
containing	NN	O	O
the	NN	O	O
-148	NN	O	B-DNA
to	NN	O	I-DNA
+60	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	O
granzyme	NN	O	B-DNA
B	NN	O	I-DNA
promoter	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
this	NN	O	O
region	NN	O	O
was	NN	O	O
the	NN	O	O
target	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
repression	NN	O	O
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
Ikaros	NN	O	B-protein
or	NN	O	O
AP-1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
granzyme	NN	O	B-DNA
B	NN	O	I-DNA
promoter	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
both	NN	O	O
sites	NN	O	O
participate	NN	O	O
in	NN	O	O
dexamethasone-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
granzyme	NN	O	B-DNA
B	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

Electromobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
revealed	NN	O	O
that	NN	O	O
dexamethasone	NN	O	O
abolished	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
Ikaros	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
reduced	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
dexamethasone	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
abrogate	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
granzyme	NN	O	I-DNA
B	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
at	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
and	NN	O	I-DNA
Ikaros	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
polycystic	NN	O	O
ovary	NN	O	O
syndrome	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
Many	NN	O	O
studies	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
possible	NN	O	O
hormonal	NN	O	O
dysregulation	NN	O	O
of	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
(	NN	O	O
HPA	NN	O	O
)	NN	O	O
axis	NN	O	O
and	NN	O	O
an	NN	O	O
increased	NN	O	O
cortisol	NN	O	O
clearance	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
polycystic	NN	O	O
ovary	NN	O	O
syndrome	NN	O	O
(	NN	O	O
PCOS	NN	O	O
)	NN	O	O
.	NN	O	O

Therefore	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor/s	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
characteristics	NN	O	O
in	NN	O	O
the	NN	O	O
developing	NN	O	O
of	NN	O	O
these	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
PCOS	NN	O	O
.	NN	O	O

METHOD	NN	O	O
:	NN	O	O
For	NN	O	O
this	NN	O	O
purpose	NN	O	O
,	NN	O	O
the	NN	O	O
number	NN	O	O
and	NN	O	O
affinity	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
peripheral	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
MNL	NN	O	O
)	NN	O	O
of	NN	O	O
10	NN	O	O
patients	NN	O	O
with	NN	O	O
PCOS	NN	O	O
and	NN	O	O
10	NN	O	O
healthy	NN	O	O
women	NN	O	O
(	NN	O	O
controls	NN	O	O
)	NN	O	O
were	NN	O	O
determined	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
There	NN	O	O
were	NN	O	O
no	NN	O	O
significant	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
(	NN	O	O
6500+/-1001	NN	O	O
sites/cell	NN	O	O
and	NN	O	O
6352+/-1697	NN	O	O
sites/cell	NN	O	O
,	NN	O	O
respectively	NN	O	O
;	NN	O	O
P	NN	O	O
>	NN	O	O
0.05	NN	O	O
)	NN	O	O
and	NN	O	O
affinity	NN	O	O
(	NN	O	O
3.93+/-0.89	NN	O	O
nM	NN	O	O
and	NN	O	O
4.49+/-0.71	NN	O	O
nM	NN	O	O
,	NN	O	O
respectively	NN	O	O
;	NN	O	O
P	NN	O	O
>	NN	O	O
0.05	NN	O	O
)	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
between	NN	O	O
the	NN	O	O
PCOS	NN	O	O
patients	NN	O	O
and	NN	O	O
the	NN	O	O
controls	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
HPA	NN	O	O
axis	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
cortisol	NN	O	O
metabolism	NN	O	O
observed	NN	O	O
in	NN	O	O
PCOS	NN	O	O
are	NN	O	O
not	NN	O	O
related	NN	O	O
to	NN	O	O
GR	NN	O	B-protein
deficiency	NN	O	O
.	NN	O	O

-DOCSTART-	O

Heterogeneous	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
lipocalin	NN	O	B-protein
NGAL	NN	O	I-protein
in	NN	O	O
primary	NN	O	O
breast	NN	O	O
cancers	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
neu	NN	O	O
oncogene-initiated	NN	O	O
rat	NN	O	O
mammary	NN	O	O
carcinomas	NN	O	O
uniquely	NN	O	O
over-express	NN	O	O
neu-related	NN	O	B-protein
lipocalin	NN	O	I-protein
(	NN	O	O
NRL	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
calycin	NN	O	B-protein
protein	NN	O	I-protein
superfamily	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
characterize	NN	O	O
the	NN	O	O
putative	NN	O	B-protein
human	NN	O	I-protein
homolog	NN	O	I-protein
of	NN	O	O
NRL	NN	O	B-protein
,	NN	O	O
neutrophil	NN	O	B-protein
gelatinase-associated	NN	O	I-protein
lipocalin	NN	O	I-protein
(	NN	O	O
NGAL	NN	O	B-protein
)	NN	O	O
.	NN	O	O

ngal	NN	O	O
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
found	NN	O	O
at	NN	O	O
moderate	NN	O	O
levels	NN	O	O
in	NN	O	O
only	NN	O	O
2	NN	O	O
of	NN	O	O
17	NN	O	O
human	NN	O	O
tissues	NN	O	O
examined	NN	O	O
,	NN	O	O
breast	NN	O	O
and	NN	O	O
lung	NN	O	O
.	NN	O	O

When	NN	O	O
breast	NN	O	O
cancers	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
NGAL	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
levels	NN	O	O
,	NN	O	O
they	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
exhibit	NN	O	O
heterogeneous	NN	O	O
expression	NN	O	O
.	NN	O	O

NGAL	NN	O	B-protein
levels	NN	O	O
varied	NN	O	O
in	NN	O	O
these	NN	O	O
tumors	NN	O	O
from	NN	O	O
undetectable	NN	O	O
to	NN	O	O
exceeding	NN	O	O
those	NN	O	O
in	NN	O	O
normal	NN	O	O
breast	NN	O	O
parenchyma	NN	O	O
.	NN	O	O

Immuno-histochemical	NN	O	O
analysis	NN	O	O
confirmed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
NGAL	NN	O	B-protein
within	NN	O	O
breast	NN	O	B-cell_type
carcinoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
detected	NN	O	O
only	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
in	NN	O	O
normal	NN	O	O
ductal	NN	O	O
epithelium	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
large	NN	O	O
amounts	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
were	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
lumen	NN	O	O
of	NN	O	O
normal	NN	O	O
breast	NN	O	O
ducts	NN	O	O
in	NN	O	O
the	NN	O	O
vicinity	NN	O	O
of	NN	O	O
NGAL	NN	O	B-protein
-expressing	NN	O	O
tumors	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
unlike	NN	O	O
NRL	NN	O	B-protein
in	NN	O	O
rat	NN	O	O
mammary	NN	O	O
carcinomas	NN	O	O
,	NN	O	O
no	NN	O	O
significant	NN	O	O
association	NN	O	O
between	NN	O	O
NGAL	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
HER-2/neu	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
human	NN	O	O
breast	NN	O	O
tumors	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
correlation	NN	O	O
between	NN	O	O
NGAL	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
was	NN	O	O
found	NN	O	O
with	NN	O	O
several	NN	O	O
other	NN	O	O
markers	NN	O	O
of	NN	O	O
poor	NN	O	O
prognosis	NN	O	O
,	NN	O	O
including	NN	O	O
estrogen	NN	O	O
and	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
-negative	NN	O	O
status	NN	O	O
and	NN	O	O
high	NN	O	O
proliferation	NN	O	O
(	NN	O	O
S-phase	NN	O	O
fraction	NN	O	O
)	NN	O	O
.	NN	O	O

NGAL	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
stratified	NN	O	O
as	NN	O	O
high	NN	O	O
or	NN	O	O
low	NN	O	O
in	NN	O	O
breast	NN	O	O
cancers	NN	O	O
from	NN	O	O
a	NN	O	O
cohort	NN	O	O
of	NN	O	O
node-positive	NN	O	O
patients	NN	O	O
with	NN	O	O
known	NN	O	O
outcome	NN	O	O
.	NN	O	O

No	NN	O	O
significant	NN	O	O
association	NN	O	O
between	NN	O	O
NGAL	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
disease-free	NN	O	O
or	NN	O	O
overall	NN	O	O
survival	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

-DOCSTART-	O

BASH	NN	O	B-protein
,	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
signaling	NN	O	I-protein
molecule	NN	O	I-protein
preferentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
bursa	NN	O	O
of	NN	O	O
Fabricius	NN	O	O
.	NN	O	O

The	NN	O	O
bursa	NN	O	O
of	NN	O	O
Fabricius	NN	O	O
is	NN	O	O
a	NN	O	O
gut-associated	NN	O	O
lymphoid	NN	O	O
organ	NN	O	O
that	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
a	NN	O	O
diversified	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
repertoire	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
chicken	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
here	NN	O	O
a	NN	O	O
novel	NN	O	O
gene	NN	O	O
preferentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
bursal	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
gene	NN	O	O
encodes	NN	O	O
an	NN	O	O
85-kDa	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
designated	NN	O	O
BASH	NN	O	B-protein
(	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
adaptor	NN	O	I-protein
containing	NN	O	I-protein
SH2	NN	O	I-protein
domain	NN	O	I-protein
)	NN	O	O
,	NN	O	O
that	NN	O	O
contains	NN	O	O
N-terminal	NN	O	B-protein
acidic	NN	O	I-protein
domains	NN	O	I-protein
with	NN	O	O
SH2	NN	O	B-protein
domain-binding	NN	O	I-protein
phosphotyrosine-based	NN	O	I-protein
motifs	NN	O	I-protein
,	NN	O	O
a	NN	O	O
proline-rich	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
C-terminal	NN	O	B-protein
SH2	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

BASH	NN	O	B-protein
shows	NN	O	O
a	NN	O	O
substantial	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
to	NN	O	O
SLP-76	NN	O	B-protein
,	NN	O	O
an	NN	O	O
adaptor	NN	O	O
protein	NN	O	O
functioning	NN	O	O
in	NN	O	O
TCR	NN	O	B-protein
-signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

BASH	NN	O	B-protein
becomes	NN	O	O
tyrosine-phosphorylated	NN	O	O
with	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
Ag	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
BCR	NN	O	B-protein
)	NN	O	O
cross-link	NN	O	O
or	NN	O	O
by	NN	O	O
coexpression	NN	O	O
with	NN	O	O
Syk	NN	O	B-protein
and	NN	O	O
Lyn	NN	O	B-protein
and	NN	O	O
associates	NN	O	O
with	NN	O	O
signaling	NN	O	B-protein
molecules	NN	O	I-protein
including	NN	O	O
Syk	NN	O	B-protein
and	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
chicken	NN	O	I-protein
Shc	NN	O	I-protein
homologue	NN	O	I-protein
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
BASH	NN	O	B-protein
results	NN	O	O
in	NN	O	O
suppression	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
BCR	NN	O	B-protein
-cross-linking	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
BASH	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
BCR	NN	O	B-protein
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
and	NN	O	O
could	NN	O	O
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
in	NN	O	O
the	NN	O	O
bursa	NN	O	O
.	NN	O	O

-DOCSTART-	O

Granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
activates	NN	O	O
a	NN	O	O
72-kDa	NN	O	B-protein
isoform	NN	O	I-protein
of	NN	O	O
STAT3	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
.	NN	O	O

Granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
G-CSF	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
involves	NN	O	O
activation	NN	O	O
of	NN	O	O
STATs	NN	O	B-protein
,	NN	O	O
proteins	NN	O	O
that	NN	O	O
serve	NN	O	O
the	NN	O	O
dual	NN	O	O
function	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
G-CSF	NN	O	B-protein
activated	NN	O	O
a	NN	O	O
distinct	NN	O	O
Stat3-like	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
immature	NN	O	B-cell_type
and	NN	O	I-cell_type
mature	NN	O	I-cell_type
normal	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
StatG	NN	O	B-protein
.	NN	O	O

StatG	NN	O	B-protein
in	NN	O	O
normal	NN	O	B-cell_type
immature	NN	O	I-cell_type
human	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
i.e	NN	O	O
.	NN	O	O
adult	NN	O	B-cell_type
CD34+	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
was	NN	O	O
composed	NN	O	O
of	NN	O	O
Stat3beta	NN	O	B-protein
.	NN	O	O

This	NN	O	O
investigation	NN	O	O
was	NN	O	O
undertaken	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
composition	NN	O	O
of	NN	O	O
StatG	NN	O	B-protein
in	NN	O	O
mature	NN	O	B-cell_type
normal	NN	O	I-cell_type
human	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
i.e	NN	O	O
.	NN	O	O
polymorphonuclear	NN	O	B-cell_type
neutrophilic	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
(	NN	O	O
PMN	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
major	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
extracts	NN	O	O
of	NN	O	O
PMN	NN	O	B-cell_type
activated	NN	O	O
by	NN	O	O
G-CSF	NN	O	B-protein
to	NN	O	O
bind	NN	O	O
the	NN	O	O
high-affinity	NN	O	B-DNA
serum-inducible	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
hSIE	NN	O	B-DNA
)	NN	O	O
is	NN	O	O
a	NN	O	O
72-kDa	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
cross-reacts	NN	O	O
with	NN	O	O
Stat3	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
,	NN	O	O
which	NN	O	O
we	NN	O	O
have	NN	O	O
designated	NN	O	O
Stat3gamma	NN	O	B-protein
.	NN	O	O

Stat3gamma	NN	O	B-protein
is	NN	O	O
derived	NN	O	O
from	NN	O	O
Stat3alpha	NN	O	B-protein
by	NN	O	O
limited	NN	O	O
proteolysis	NN	O	O
and	NN	O	O
lacks	NN	O	O
the	NN	O	O
carboxyl-terminal	NN	O	B-protein
portion	NN	O	I-protein
of	NN	O	O
Stat3alpha	NN	O	B-protein
.	NN	O	O

Because	NN	O	O
this	NN	O	O
region	NN	O	O
of	NN	O	O
Stat3alpha	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
Stat3gamma	NN	O	B-protein
may	NN	O	O
be	NN	O	O
transcriptionally	NN	O	O
inactive	NN	O	O
and	NN	O	O
may	NN	O	O
compete	NN	O	O
with	NN	O	O
Stat3alpha	NN	O	B-protein
for	NN	O	O
Stat3	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
these	NN	O	O
terminally	NN	O	B-cell_type
differentiated	NN	O	I-cell_type
myeloid	NN	O	I-cell_type
cells	NN	O	I-cell_type

-DOCSTART-	O

Association	NN	O	O
between	NN	O	O
expression	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
and	NN	O	O
integration	NN	O	O
of	NN	O	O
human	NN	O	O
T-cell-leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
in	NN	O	O
adult	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
is	NN	O	O
known	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
levels	NN	O	O
of	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
in	NN	O	O
adult	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
(	NN	O	I-cell_type
ATL	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
high	NN	O	O
,	NN	O	O
whereas	NN	O	O
those	NN	O	O
in	NN	O	O
T-lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
not	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
factors	NN	O	O
that	NN	O	O
influence	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
molecules	NN	O	O
,	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
to	NN	O	O
measure	NN	O	O
the	NN	O	O
expression	NN	O	O
level	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-RNA
mRNAs	NN	O	I-RNA
and	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
integration	NN	O	O
of	NN	O	O
human	NN	O	O
T-cell-leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
provirus	NN	O	O
were	NN	O	O
done	NN	O	O
.	NN	O	O

The	NN	O	O
levels	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
of	NN	O	O
ATL	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
generally	NN	O	O
higher	NN	O	O
than	NN	O	O
those	NN	O	O
of	NN	O	O
T-lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
ILT-mat	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
ATL16T	NN	O	B-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
they	NN	O	O
were	NN	O	O
ATL	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
showed	NN	O	O
rather	NN	O	O
low	NN	O	O
surface	NN	O	O
ICAM-1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
ICAM-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
showed	NN	O	O
that	NN	O	O
only	NN	O	O
two	NN	O	O
and	NN	O	O
four	NN	O	O
bands	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
ILT-mat	NN	O	B-cell_line
and	NN	O	O
ATL16T	NN	O	B-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
whereas	NN	O	O
>	NN	O	O
10	NN	O	O
bands	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
other	NN	O	O
ATL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
monoclonal	NN	O	O
integration	NN	O	O
of	NN	O	O
HTLV-1	NN	O	O
provirus	NN	O	O
to	NN	O	O
the	NN	O	O
genome	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
,	NN	O	O
especially	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
integration	NN	O	O
sites	NN	O	O
,	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
factors	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Heat-shock	NN	O	O
and	NN	O	O
cadmium	NN	O	O
chloride	NN	O	O
increase	NN	O	O
the	NN	O	O
vimentin	NN	O	O
mRNA	NN	O	O
and	NN	O	O
protein	NN	O	O
levels	NN	O	O
in	NN	O	O
U-937	NN	O	B-cell_line
human	NN	O	I-cell_line
promonocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Heat-shock	NN	O	O
for	NN	O	O
2	NN	O	O
hours	NN	O	O
at	NN	O	O
42	NN	O	O
degrees	NN	O	O
C	NN	O	O
,	NN	O	O
or	NN	O	O
the	NN	O	O
administration	NN	O	O
for	NN	O	O
3	NN	O	O
hours	NN	O	O
of	NN	O	O
100	NN	O	O
or	NN	O	O
150	NN	O	O
microM	NN	O	O
cadmium	NN	O	O
chloride	NN	O	O
,	NN	O	O
inhibited	NN	O	O
the	NN	O	O
subsequent	NN	O	B-cell_line
proliferation	NN	O	I-cell_line
activity	NN	O	I-cell_line
,	NN	O	O
induced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
functional	NN	O	O
differentiation	NN	O	B-protein
markers	NN	O	I-protein
,	NN	O	O
and	NN	O	O
caused	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
the	NN	O	O
stress-responsive	NN	O	B-protein
HSP70	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
U-937	NN	O	B-cell_line
human	NN	O	I-cell_line
promonocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
both	NN	O	O
heat	NN	O	O
and	NN	O	O
cadmium	NN	O	O
produced	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
the	NN	O	O
intermediate	NN	O	B-protein
filament	NN	O	I-protein
protein	NN	O	I-protein
vimentin	NN	O	B-protein
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
immunoblot	NN	O	O
and	NN	O	O
immunofluorescence	NN	O	O
assays	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
amounts	NN	O	O
of	NN	O	O
actin	NN	O	B-protein
and	NN	O	O
beta-tubulin	NN	O	B-protein
were	NN	O	O
not	NN	O	O
significantly	NN	O	O
altered	NN	O	O
.	NN	O	O

The	NN	O	O
amount	NN	O	O
of	NN	O	O
vimentin	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
also	NN	O	O
increased	NN	O	O
during	NN	O	O
recovery	NN	O	O
from	NN	O	O
stress	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
vimentin	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
not	NN	O	O
exclusively	NN	O	O
regulated	NN	O	O
at	NN	O	O
the	NN	O	O
protein	NN	O	O
level	NN	O	O
.	NN	O	O

Although	NN	O	O
cadmium	NN	O	O
caused	NN	O	O
an	NN	O	O
early	NN	O	O
,	NN	O	O
transient	NN	O	O
stimulation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
heat-shock	NN	O	O
failed	NN	O	O
to	NN	O	O
alter	NN	O	O
both	NN	O	O
protooncogene	NN	O	B-DNA
expression	NN	O	O
and	NN	O	O
transcription	NN	O	O
factor	NN	O	O
binding	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
stress-induced	NN	O	O
vimentin	NN	O	B-protein
increase	NN	O	O
was	NN	O	O
not	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
-mediated	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
observed	NN	O	O
that	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
decay	NN	O	O
of	NN	O	O
vimentin	NN	O	B-RNA
mRNA	NN	O	I-RNA
upon	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
administration	NN	O	O
was	NN	O	O
decreased	NN	O	O
in	NN	O	O
heat-	NN	O	B-cell_line
and	NN	O	I-cell_line
cadmium-pretreated	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
comparison	NN	O	O
to	NN	O	O
untreated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
stress	NN	O	O
treatments	NN	O	O
cause	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
vimentin	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
promonocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
explained	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
transcript	NN	O	O
stabilization	NN	O	O
.	NN	O	O

-DOCSTART-	O

Ovarian	NN	O	O
and	NN	O	O
breast	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
can	NN	O	O
recognize	NN	O	O
peptides	NN	O	O
from	NN	O	O
the	NN	O	O
amino	NN	O	O
enhancer	NN	O	O
of	NN	O	O
split	NN	O	O
protein	NN	O	O
of	NN	O	O
the	NN	O	O
Notch	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
investigated	NN	O	O
recognition	NN	O	O
by	NN	O	O
ovarian	NN	O	B-cell_type
tumor	NN	O	I-cell_type
associated	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
(	NN	O	O
OVTAL	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
and	NN	O	O
breast	NN	O	B-cell_type
tumor	NN	O	I-cell_type
associated	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
BRTAL	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
of	NN	O	O
peptides	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
sequence	NN	O	O
125-135	NN	O	B-protein
of	NN	O	I-protein
the	NN	O	I-protein
Aminoenhancer	NN	O	I-protein
of	NN	O	I-protein
split	NN	O	I-protein
(	NN	O	I-protein
AES	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Three	NN	O	O
of	NN	O	O
these	NN	O	O
peptides	NN	O	O
designated	NN	O	O
as	NN	O	O
G75	NN	O	O
:	NN	O	O
AES1/2	NN	O	O
(	NN	O	O
128-135	NN	O	B-protein
)	NN	O	O
,	NN	O	O
G60	NN	O	O
:	NN	O	O
AES1/2	NN	O	O
(	NN	O	O
127-137	NN	O	B-protein
)	NN	O	O
and	NN	O	O
G61	NN	O	O
:	NN	O	O
AES1/2	NN	O	O
(	NN	O	O
125-133	NN	O	B-protein
)	NN	O	O
correspond	NN	O	O
to	NN	O	O
the	NN	O	O
wildtype	NN	O	B-DNA
AES	NN	O	I-DNA
sequence	NN	O	I-DNA
,	NN	O	O
while	NN	O	O
the	NN	O	O
fourth	NN	O	O
G76	NN	O	O
:	NN	O	O
GPLTPLPV	NN	O	O
,	NN	O	O
AES1/2	NN	O	B-protein
(	NN	O	I-protein
128-135	NN	O	I-protein
)	NN	O	I-protein
corresponds	NN	O	O
to	NN	O	O
a	NN	O	O
variant	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
peptide	NN	O	O
G75	NN	O	O
with	NN	O	O
the	NN	O	O
N-terminal	NN	O	O
Leu	NN	O	O
substituted	NN	O	O
to	NN	O	O
glycine	NN	O	O
.	NN	O	O

These	NN	O	O
sequences	NN	O	O
were	NN	O	O
chosen	NN	O	O
for	NN	O	O
study	NN	O	O
because	NN	O	O
mass-spectrometric	NN	O	O
analysis	NN	O	O
(	NN	O	O
MS	NN	O	O
)	NN	O	O
of	NN	O	O
a	NN	O	O
CTL	NN	O	B-cell_type
active	NN	O	O
HPLC	NN	O	O
peptide	NN	O	O
fraction	NN	O	O
eluted	NN	O	O
from	NN	O	O
immunoaffinity	NN	O	B-protein
precipitated	NN	O	I-protein
HLA-A2	NN	O	I-protein
molecule	NN	O	I-protein
,	NN	O	O
revealed	NN	O	O
:	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
ion	NN	O	O
with	NN	O	O
a	NN	O	O
mass-to-charge	NN	O	O
ratio	NN	O	O
(	NN	O	O
m/z	NN	O	O
)	NN	O	O
of	NN	O	O
793	NN	O	O
which	NN	O	O
was	NN	O	O
more	NN	O	O
abundant	NN	O	O
than	NN	O	O
other	NN	O	O
ions	NN	O	O
of	NN	O	O
similar	NN	O	O
masses	NN	O	O
;	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
the	NN	O	O
tentatively	NN	O	O
reconstituted	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
ion	NN	O	O
793	NN	O	O
matched	NN	O	O
the	NN	O	O
sequence	NN	O	O
of	NN	O	O
peptide	NN	O	O
G76	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
AES	NN	O	O
peptides	NN	O	O
G75	NN	O	O
(	NN	O	O
128-135	NN	O	B-protein
)	NN	O	O
and	NN	O	O
G76	NN	O	O
(	NN	O	O
128-135	NN	O	B-protein
)	NN	O	O
(	NN	O	O
L128G	NN	O	O
)	NN	O	O
reconstituted	NN	O	O
CTL	NN	O	O
recognition	NN	O	O
at	NN	O	O
concentrations	NN	O	O
ranging	NN	O	O
between	NN	O	O
200-500	NN	O	O
nM	NN	O	O
.	NN	O	O

These	NN	O	O
concentrations	NN	O	O
are	NN	O	O
lower	NN	O	O
than	NN	O	O
concentrations	NN	O	O
reported	NN	O	O
to	NN	O	O
activate	NN	O	O
effector	NN	O	O
function	NN	O	O
of	NN	O	O
CTL	NN	O	O
recognizing	NN	O	O
other	NN	O	O
epithelial	NN	O	B-protein
tumor	NN	O	I-protein
Ag	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
analysis	NN	O	O
with	NN	O	O
cloned	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
indicated	NN	O	O
that	NN	O	O
G75	NN	O	O
and	NN	O	O
G76	NN	O	O
were	NN	O	O
not	NN	O	O
cross-reactive	NN	O	O
specificities	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
residues	NN	O	I-protein
of	NN	O	O
the	NN	O	O
variant	NN	O	O
peptide	NN	O	O
in	NN	O	O
dictating	NN	O	O
specificities	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
AES	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
part	NN	O	O
of	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
repressors	NN	O	O
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
Enhancer	NN	O	B-DNA
of	NN	O	I-DNA
split	NN	O	I-DNA
[	NN	O	I-DNA
E	NN	O	I-DNA
(	NN	O	I-DNA
spl	NN	O	I-DNA
)	NN	O	I-DNA
]	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
since	NN	O	O
these	NN	O	O
repressors	NN	O	O
are	NN	O	O
activated	NN	O	O
to	NN	O	O
suppress	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
Notch	NN	O	B-protein
receptors	NN	O	I-protein
signalling	NN	O	O
,	NN	O	O
the	NN	O	O
AES	NN	O	O
peptides	NN	O	O
may	NN	O	O
represent	NN	O	O
a	NN	O	O
novel	NN	O	O
class	NN	O	O
of	NN	O	O
self-antigens	NN	O	B-protein
that	NN	O	O
deserve	NN	O	O
further	NN	O	O
consideration	NN	O	O
as	NN	O	O
tumor	NN	O	B-protein
Ag	NN	O	I-protein
in	NN	O	O
epithelial	NN	O	O
cancers	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
the	NN	O	O
pro-myelocytic	NN	O	B-DNA
leukaemia	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
interferons	NN	O	I-protein
.	NN	O	O

The	NN	O	O
physiological	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
pro-myelocytic	NN	O	B-protein
leukaemia	NN	O	I-protein
(	NN	O	I-protein
PML	NN	O	I-protein
)	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
is	NN	O	O
poorly	NN	O	O
defined	NN	O	O
.	NN	O	O

Among	NN	O	O
other	NN	O	O
functions	NN	O	O
,	NN	O	O
PML	NN	O	B-DNA
is	NN	O	O
involved	NN	O	O
in	NN	O	O
haematopoietic	NN	O	O
differentiation	NN	O	O
and	NN	O	O
in	NN	O	O
control	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
tumorigenesis	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	O
PML	NN	O	B-DNA
expression	NN	O	O
by	NN	O	O
interferons	NN	O	B-protein
(	NN	O	O
IFNs	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IL-1	NN	O	B-protein
in	NN	O	O
various	NN	O	O
human	NN	O	B-cell_line
haematopoietic	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
U937	NN	O	B-cell_line
,	NN	O	O
THP1	NN	O	B-cell_line
,	NN	O	O
HL60	NN	O	B-cell_line
,	NN	O	O
NB4	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
diploid	NN	O	I-cell_type
fibroblasts	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

Cytokine-induced	NN	O	O
modulation	NN	O	O
of	NN	O	O
PML	NN	O	B-DNA
expression	NN	O	O
was	NN	O	O
assessed	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analyses	NN	O	O
,	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
studies	NN	O	O
and	NN	O	O
in	NN	O	O
situ	NN	O	O
immunolabelling	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
IFNs	NN	O	B-protein
and	NN	O	O
IL-1	NN	O	B-protein
upregulate	NN	O	O
PML	NN	O	O
transcript	NN	O	O
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
in	NN	O	O
a	NN	O	O
time	NN	O	O
and	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

In	NN	O	O
situ	NN	O	O
immunolabelling	NN	O	O
revealed	NN	O	O
that	NN	O	O
upregulation	NN	O	O
of	NN	O	O
protein	NN	O	O
expression	NN	O	O
by	NN	O	O
IFN-alpha	NN	O	B-protein
is	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
a	NN	O	O
marked	NN	O	O
increase	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
number	NN	O	O
and	NN	O	O
the	NN	O	O
intensity	NN	O	O
of	NN	O	O
the	NN	O	O
staining	NN	O	O
of	NN	O	O
so-called	NN	O	O
PML	NN	O	B-protein
nuclear	NN	O	I-protein
bodies	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
PML	NN	O	B-DNA
expression	NN	O	O
by	NN	O	O
interferons	NN	O	B-protein
and	NN	O	O
IL-1	NN	O	B-protein
may	NN	O	O
account	NN	O	O
for	NN	O	O
upregulation	NN	O	O
of	NN	O	O
PML	NN	O	B-protein
proteins	NN	O	I-protein
observed	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
tissues	NN	O	O
and	NN	O	O
in	NN	O	O
proliferative	NN	O	O
states	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
ZEBRA	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
does	NN	O	O
not	NN	O	O
influence	NN	O	O
its	NN	O	O
biological	NN	O	O
activity	NN	O	O
.	NN	O	O

Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
which	NN	O	O
latently	NN	O	O
infects	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

EBV	NN	O	O
encodes	NN	O	O
a	NN	O	O
unique	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
,	NN	O	O
known	NN	O	O
as	NN	O	O
ZEBRA	NN	O	B-protein
,	NN	O	O
which	NN	O	O
can	NN	O	O
disrupt	NN	O	O
viral	NN	O	O
latency	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
induce	NN	O	O
lytic	NN	O	O
viral	NN	O	O
replication	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
ZEBRA	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
at	NN	O	O
the	NN	O	O
EBV	NN	O	O
origin	NN	O	O
of	NN	O	O
lytic	NN	O	O
replication	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
viral	NN	O	O
DNA	NN	O	O
replication	NN	O	O
to	NN	O	O
occur	NN	O	O
.	NN	O	O

Previously	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
heterologous	NN	O	B-protein
activation	NN	O	I-protein
domains	NN	O	I-protein
can	NN	O	O
fully	NN	O	O
substitute	NN	O	O
for	NN	O	O
the	NN	O	O
ZEBRA	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
extend	NN	O	O
those	NN	O	O
results	NN	O	O
by	NN	O	O
showing	NN	O	O
that	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
ZEBRA	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
or	NN	O	O
a	NN	O	O
heterologous	NN	O	O
replacement	NN	O	O
domain	NN	O	O
does	NN	O	O
not	NN	O	O
influence	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
function	NN	O	O
in	NN	O	O
the	NN	O	O
disruption	NN	O	O
of	NN	O	O
EBV	NN	O	O
latency	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
three	NN	O	O
novel	NN	O	O
clones	NN	O	O
were	NN	O	O
constructed	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
ZEBRA	NN	O	B-protein
activation	NN	O	I-protein
region	NN	O	I-protein
was	NN	O	O
repositioned	NN	O	O
to	NN	O	O
the	NN	O	O
carboxy	NN	O	B-protein
terminus	NN	O	I-protein
of	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

These	NN	O	O
mutants	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-protein
's	NN	O	I-protein
wild	NN	O	I-protein
type	NN	O	I-protein
domain	NN	O	I-protein
to	NN	O	O
function	NN	O	O
in	NN	O	O
the	NN	O	O
complex	NN	O	O
biological	NN	O	O
process	NN	O	O
of	NN	O	O
virus	NN	O	O
activation	NN	O	O
is	NN	O	O
not	NN	O	O
compromised	NN	O	O
by	NN	O	O
altering	NN	O	O
its	NN	O	O
position	NN	O	O
within	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
modulation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
content	NN	O	O
by	NN	O	O
3-O-methyl-D-glucose	NN	O	O
transport	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocyte	NN	O	I-cell_type
in	NN	O	O
obesity	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
3-O-methyl-D	NN	O	O
glucose	NN	O	O
(	NN	O	O
3-O-MG	NN	O	O
)	NN	O	O
transport	NN	O	O
were	NN	O	O
determined	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
MNL	NN	O	B-cell_type
)	NN	O	O
from	NN	O	O
11	NN	O	O
abdominal	NN	O	O
obese	NN	O	O
subjects	NN	O	O
,	NN	O	O
10	NN	O	O
pituitary-dependent	NN	O	O
Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
(	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
)	NN	O	O
and	NN	O	O
10	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
whole-cell	NN	O	O
competitive	NN	O	O
binding	NN	O	O
assay	NN	O	O
and	NN	O	O
3H-dexamethasone	NN	O	O
as	NN	O	O
tracer	NN	O	O
,	NN	O	O
MNL	NN	O	B-cell_type
of	NN	O	O
abdominal	NN	O	O
obese	NN	O	O
subjects	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
4855	NN	O	O
+/-	NN	O	O
1389	NN	O	O
sites/cell	NN	O	O
which	NN	O	O
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
than	NN	O	O
controls	NN	O	O
(	NN	O	O
6234	NN	O	O
+/-	NN	O	O
1568	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
,	NN	O	O
although	NN	O	O
no	NN	O	O
significant	NN	O	O
difference	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
mean	NN	O	O
serum	NN	O	O
cortisol	NN	O	O
level	NN	O	O
.	NN	O	O

Their	NN	O	O
mean	NN	O	O
Kd	NN	O	O
(	NN	O	O
affinity	NN	O	O
)	NN	O	O
was	NN	O	O
also	NN	O	O
significantly	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
healthy	NN	O	O
controls	NN	O	O
(	NN	O	O
obese	NN	O	O
Kd	NN	O	O
:	NN	O	O
2.92	NN	O	O
+/-	NN	O	O
0.84	NN	O	O
nmol/l	NN	O	O
,	NN	O	O
control	NN	O	O
Kd	NN	O	O
:	NN	O	O
4.55	NN	O	O
+/-	NN	O	O
0.67	NN	O	O
nM	NN	O	O
,	NN	O	O
p	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
the	NN	O	O
receptor	NN	O	O
characteristics	NN	O	O
in	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
patients	NN	O	O
were	NN	O	O
within	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
same	NN	O	O
time	NN	O	O
,	NN	O	O
3-O-MG	NN	O	O
transport	NN	O	O
was	NN	O	O
determined	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
subjects	NN	O	O
.	NN	O	O

In	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
,	NN	O	O
3-O-MG	NN	O	O
transport	NN	O	O
was	NN	O	O
within	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
abdominal	NN	O	O
obesity	NN	O	O
this	NN	O	O
value	NN	O	O
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
than	NN	O	O
the	NN	O	O
healthy	NN	O	O
controls	NN	O	O
(	NN	O	O
abdominal	NN	O	O
obese	NN	O	O
:	NN	O	O
31.90	NN	O	O
+/-	NN	O	O
8.20	NN	O	O
;	NN	O	O
control	NN	O	O
:	NN	O	O
46.26	NN	O	O
+/-	NN	O	O
12.91	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cell	NN	O	O
,	NN	O	O
min	NN	O	O
,	NN	O	O
p	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
found	NN	O	O
a	NN	O	O
positive	NN	O	O
correlation	NN	O	O
between	NN	O	O
3-O-MG	NN	O	O
transport	NN	O	O
and	NN	O	O
GR	NN	O	B-protein
binding	NN	O	O
capacity	NN	O	O
in	NN	O	O
abdominal	NN	O	O
subjects	NN	O	O
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.89	NN	O	O
,	NN	O	O
p	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
,	NN	O	O
however	NN	O	O
we	NN	O	O
did	NN	O	O
not	NN	O	O
find	NN	O	O
such	NN	O	O
a	NN	O	O
correlation	NN	O	O
in	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.60	NN	O	O
,	NN	O	O
p	NN	O	O
>	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicated	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
abdominal	NN	O	O
obesity	NN	O	O
,	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
binding	NN	O	O
capacity	NN	O	O
in	NN	O	O
MNL	NN	O	B-cell_type
is	NN	O	O
influenced	NN	O	O
by	NN	O	O
the	NN	O	O
changes	NN	O	O
in	NN	O	O
glucose	NN	O	O
transport	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin-10	NN	O	B-protein
and	NN	O	O
transforming	NN	O	B-DNA
growth	NN	O	I-DNA
factor-beta	NN	O	I-DNA
promoter	NN	O	I-DNA
polymorphisms	NN	O	I-DNA
in	NN	O	O
allergies	NN	O	O
and	NN	O	O
asthma	NN	O	O
.	NN	O	O

Interleukin-10	NN	O	B-protein
(	NN	O	O
IL-10	NN	O	B-protein
)	NN	O	O
and	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
beta	NN	O	I-protein
(	NN	O	O
TGF-beta	NN	O	B-protein
)	NN	O	O
are	NN	O	O
inhibitory	NN	O	O
for	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
IgE	NN	O	B-protein
production	NN	O	O
,	NN	O	O
and	NN	O	O
mast	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
and	NN	O	O
they	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
cytokines	NN	O	B-protein
are	NN	O	O
therefore	NN	O	O
candidate	NN	O	O
genes	NN	O	O
which	NN	O	O
could	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
asthma	NN	O	O
or	NN	O	O
allergies	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
polymorphic	NN	O	O
nucleotides	NN	O	O
within	NN	O	O
the	NN	O	O
IL-10	NN	O	B-DNA
and	NN	O	I-DNA
TGF-beta	NN	O	I-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
would	NN	O	O
link	NN	O	O
to	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
allergies	NN	O	O
and	NN	O	O
asthma	NN	O	O
.	NN	O	O

DNA	NN	O	O
taken	NN	O	O
from	NN	O	O
families	NN	O	O
with	NN	O	O
an	NN	O	O
asthmatic	NN	O	O
proband	NN	O	O
was	NN	O	O
examined	NN	O	O
for	NN	O	O
base	NN	O	O
exchanges	NN	O	O
by	NN	O	O
single-stranded	NN	O	O
conformational	NN	O	O
polymorphism	NN	O	O
(	NN	O	O
SSCP	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
polymorphism	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-10	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
four	NN	O	O
in	NN	O	O
the	NN	O	O
TGF-beta	NN	O	B-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
(	NN	O	O
3	NN	O	O
in	NN	O	O
TGF-beta1	NN	O	B-DNA
and	NN	O	O
1	NN	O	O
in	NN	O	O
TGF-beta2	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

The	NN	O	O
IL-10	NN	O	B-protein
gene	NN	O	O
polymorphism	NN	O	O
was	NN	O	O
a	NN	O	O
C-to-A	NN	O	O
exchange	NN	O	O
571	NN	O	B-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
upstream	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
translation	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
was	NN	O	O
present	NN	O	O
between	NN	O	O
consensus	NN	O	B-DNA
binding	NN	O	I-DNA
sequences	NN	O	I-DNA
for	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
elevated	NN	O	O
total	NN	O	O
serum	NN	O	O
.	NN	O	O

This	NN	O	O
polymorphism	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
elevated	NN	O	O
total	NN	O	B-protein
serum	NN	O	I-protein
IgE	NN	O	I-protein
in	NN	O	O
subjects	NN	O	O
heterozygotic	NN	O	O
or	NN	O	O
homozygotic	NN	O	O
for	NN	O	O
this	NN	O	O
base	NN	O	O
exchange	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.009	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
base	NN	O	O
exchange	NN	O	O
at	NN	O	O
-509	NN	O	O
(	NN	O	O
from	NN	O	O
the	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
site	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
TGF-beta	NN	O	B-DNA
promoter	NN	O	I-DNA
also	NN	O	O
linked	NN	O	O
to	NN	O	O
elevated	NN	O	O
total	NN	O	O
IgE	NN	O	B-protein
(	NN	O	O
p	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
polymorphism	NN	O	O
represented	NN	O	O
a	NN	O	O
C-to-T	NN	O	O
base	NN	O	O
exchange	NN	O	O
which	NN	O	O
induced	NN	O	O
a	NN	O	O
YY1	NN	O	B-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
associated	NN	O	O
with	NN	O	O
negative	NN	O	O
transcription	NN	O	O
regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tpl-2	NN	O	B-protein
induces	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
by	NN	O	O
triggering	NN	O	O
multiple	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
that	NN	O	O
activate	NN	O	O
NFAT	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
Tpl-2	NN	O	B-protein
kinase	NN	O	I-protein
activates	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
and	NN	O	O
induces	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
Tpl-2	NN	O	B-protein
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
mutant	NN	O	B-protein
signaling	NN	O	I-protein
molecules	NN	O	I-protein
that	NN	O	O
inhibit	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
or	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
/NFAT	NN	O	B-protein
pathways	NN	O	O
and	NN	O	O
is	NN	O	O
promoted	NN	O	O
by	NN	O	O
combinations	NN	O	O
of	NN	O	O
signaling	NN	O	B-protein
molecules	NN	O	I-protein
that	NN	O	O
activate	NN	O	O
these	NN	O	O
pathways	NN	O	O
.	NN	O	O

We	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
conclude	NN	O	O
that	NN	O	O
signals	NN	O	O
generated	NN	O	O
by	NN	O	O
the	NN	O	O
convergence	NN	O	O
of	NN	O	O
the	NN	O	O
MAPK	NN	O	B-protein
and	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
/NFAT	NN	O	B-protein
pathway	NN	O	O
are	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
Tpl-2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
an	NN	O	O
NFAT-driven	NN	O	B-DNA
minimal	NN	O	I-DNA
promoter	NN	O	I-DNA
were	NN	O	O
shown	NN	O	O
to	NN	O	O
depend	NN	O	O
on	NN	O	O
signals	NN	O	O
transduced	NN	O	O
by	NN	O	O
Raf1	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
only	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
whose	NN	O	O
activation	NN	O	O
by	NN	O	O
Tpl-2	NN	O	B-protein
was	NN	O	O
fully	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
mutant	NN	O	I-protein
MEK1S218/222A	NN	O	I-protein
and	NN	O	O
the	NN	O	O
MEK1/MEK2	NN	O	O
inhibitor	NN	O	O
PD098059	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
is	NN	O	O
MAPK	NN	O	B-protein
-dependent	NN	O	O
these	NN	O	O
findings	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
MAPK	NN	O	B-protein
by	NN	O	O
Tpl-2	NN	O	B-protein
is	NN	O	O
either	NN	O	O
independent	NN	O	O
or	NN	O	O
only	NN	O	O
partially	NN	O	O
dependent	NN	O	O
on	NN	O	O
MEK1	NN	O	B-protein
and	NN	O	O
MEK2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
they	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
not	NN	O	O
only	NN	O	O
NFAT	NN	O	B-protein
but	NN	O	O
also	NN	O	O
a	NN	O	O
second	NN	O	O
factor	NN	O	O
whose	NN	O	O
activation	NN	O	O
is	NN	O	O
MEK	NN	O	B-protein
-dependent	NN	O	O
.	NN	O	O

Experiments	NN	O	O
in	NN	O	O
COS-1	NN	O	B-cell_line
and	NN	O	O
EL-4	NN	O	B-cell_line
cells	NN	O	I-cell_line
confirmed	NN	O	O
both	NN	O	O
hypotheses	NN	O	O
and	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
second	NN	O	O
factor	NN	O	O
activated	NN	O	O
by	NN	O	O
Tpl-2	NN	O	B-protein
is	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

While	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
an	NN	O	O
NFAT-driven	NN	O	B-DNA
minimal	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
Tpl-2	NN	O	B-protein
was	NN	O	O
fully	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
mutant	NN	O	I-protein
NFAT	NN	O	B-protein
delta418	NN	O	I-protein
,	NN	O	O
it	NN	O	O
was	NN	O	O
only	NN	O	O
partially	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
inhibitor	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
Tpl-2	NN	O	B-protein
-mediated	NN	O	O
NFAT	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
calcineurin-dependent	NN	O	O
and	NN	O	O
independent	NN	O	O
pathways	NN	O	O
.	NN	O	O

Both	NN	O	O
pathways	NN	O	O
were	NN	O	O
fully	NN	O	O
blocked	NN	O	O
by	NN	O	O
Bcl-2	NN	O	B-protein
or	NN	O	O
Bcl-X	NN	O	B-protein
(	NN	O	I-protein
L	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
activation	NN	O	O
of	NN	O	O
functionally	NN	O	O
distinct	NN	O	O
STAT5	NN	O	B-protein
proteins	NN	O	I-protein
by	NN	O	O
IL-5	NN	O	B-protein
and	NN	O	O
GM-CSF	NN	O	B-protein
during	NN	O	O
eosinophil	NN	O	O
and	NN	O	O
neutrophil	NN	O	O
differentiation	NN	O	O
from	NN	O	O
human	NN	O	B-cell_line
CD34+	NN	O	I-cell_line
hematopoietic	NN	O	I-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Interleukin-5	NN	O	B-protein
(	NN	O	O
IL-5	NN	O	B-protein
)	NN	O	O
and	NN	O	O
granulocyte	NN	O	B-protein
macrophage-colony	NN	O	I-protein
stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
are	NN	O	O
important	NN	O	O
cytokines	NN	O	B-protein
for	NN	O	O
the	NN	O	O
proliferation	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
JAK	NN	O	B-protein
/STAT	NN	O	B-protein
pathway	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
implicated	NN	O	O
in	NN	O	O
mediating	NN	O	O
biological	NN	O	O
responses	NN	O	O
induced	NN	O	O
by	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
predominantly	NN	O	O
activate	NN	O	O
an	NN	O	O
80	NN	O	B-protein
kDa	NN	O	I-protein
STAT5	NN	O	I-protein
isoform	NN	O	I-protein
in	NN	O	O
mature	NN	O	O
granulocytes	NN	O	O
.	NN	O	O

To	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
STAT	NN	O	O
proteins	NN	O	O
during	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
granulocytes	NN	O	O
,	NN	O	O
we	NN	O	O
evaluated	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
CD34+	NN	O	I-cell_line
hematopoietic	NN	O	I-cell_line
stem	NN	O	I-cell_line
cells	NN	O	I-cell_line
ex	NN	O	O
vivo	NN	O	O
toward	NN	O	O
eosinophils	NN	O	B-cell_type
and	NN	O	O
neutrophils	NN	O	O
.	NN	O	O

Bandshift	NN	O	O
experiments	NN	O	O
showed	NN	O	O
that	NN	O	O
in	NN	O	O
an	NN	O	O
early	NN	O	O
stage	NN	O	O
of	NN	O	O
both	NN	O	O
differentiation	NN	O	O
pathways	NN	O	O
(	NN	O	O
14	NN	O	O
days	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
94	NN	O	O
kDa	NN	O	O
STAT5B	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
activated	NN	O	O
by	NN	O	O
both	NN	O	O
IL-5	NN	O	B-protein
(	NN	O	O
eosinophil	NN	O	B-cell_type
lineage	NN	O	I-cell_type
)	NN	O	O
and	NN	O	O
GM-CSF	NN	O	B-protein
(	NN	O	O
neutrophil	NN	O	B-cell_type
lineage	NN	O	I-cell_type
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
during	NN	O	O
maturation	NN	O	O
of	NN	O	O
both	NN	O	O
lineages	NN	O	O
(	NN	O	O
days	NN	O	O
21	NN	O	O
and	NN	O	O
28	NN	O	O
)	NN	O	O
,	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
functionally	NN	O	O
distinct	NN	O	O
80	NN	O	B-protein
kDa	NN	O	I-protein
STAT5	NN	O	I-protein
isoform	NN	O	I-protein
was	NN	O	O
observed	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
heterodimer	NN	O	B-protein
DNA-binding	NN	O	I-protein
complexes	NN	O	I-protein
containing	NN	O	O
both	NN	O	O
the	NN	O	O
94	NN	O	O
and	NN	O	O
80	NN	O	O
kDa	NN	O	O
STAT5	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
finding	NN	O	O
that	NN	O	O
functionally	NN	O	O
distinct	NN	O	O
isoforms	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
are	NN	O	O
activated	NN	O	O
during	NN	O	O
the	NN	O	O
early	NN	O	O
and	NN	O	O
late	NN	O	O
differentiation	NN	O	O
stages	NN	O	O
of	NN	O	O
granulocytes	NN	O	O
suggests	NN	O	O
that	NN	O	O
they	NN	O	O
might	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
regulating	NN	O	O
different	NN	O	O
biological	NN	O	O
functions	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
T-cell	NN	O	B-protein
oncogenic	NN	O	I-protein
protein	NN	O	I-protein
HOX11	NN	O	I-protein
activates	NN	O	O
Aldh1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
NIH	NN	O	B-cell_line
3T3	NN	O	I-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
represses	NN	O	O
its	NN	O	O
expression	NN	O	O
in	NN	O	O
mouse	NN	O	O
spleen	NN	O	O
development	NN	O	O
.	NN	O	O

Hox11	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
homeobox	NN	O	B-DNA
gene	NN	O	I-DNA
essential	NN	O	O
for	NN	O	O
spleen	NN	O	O
formation	NN	O	O
in	NN	O	O
mice	NN	O	O
,	NN	O	O
since	NN	O	O
atrophy	NN	O	O
of	NN	O	O
the	NN	O	O
anlage	NN	O	O
of	NN	O	O
a	NN	O	O
developing	NN	O	O
spleen	NN	O	O
occurs	NN	O	O
in	NN	O	O
early	NN	O	O
embryonic	NN	O	O
development	NN	O	O
in	NN	O	O
Hox11	NN	O	O
null	NN	O	O
mice	NN	O	O
.	NN	O	O

HOX11	NN	O	B-protein
is	NN	O	O
also	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
T-cell	NN	O	O
acute	NN	O	O
leukemias	NN	O	O
after	NN	O	O
specific	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
protein	NN	O	O
has	NN	O	O
a	NN	O	O
homeodomain	NN	O	B-DNA
and	NN	O	O
can	NN	O	O
activate	NN	O	O
transcription	NN	O	O
,	NN	O	O
it	NN	O	O
probably	NN	O	O
exerts	NN	O	O
at	NN	O	O
least	NN	O	O
some	NN	O	O
of	NN	O	O
its	NN	O	O
effects	NN	O	O
in	NN	O	O
vivo	NN	O	O
by	NN	O	O
regulation	NN	O	O
of	NN	O	O
target	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Representational	NN	O	O
difference	NN	O	O
analysis	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
isolate	NN	O	O
cDNA	NN	O	B-DNA
clones	NN	O	I-DNA
corresponding	NN	O	O
to	NN	O	O
mRNA	NN	O	B-RNA
species	NN	O	I-RNA
activated	NN	O	O
following	NN	O	O
stable	NN	O	O
expression	NN	O	O
of	NN	O	O
HOX11	NN	O	B-protein
in	NN	O	O
NIH	NN	O	B-cell_line
3T3	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
retinoic	NN	O	B-protein
acid-synthesizing	NN	O	I-protein
enzyme	NN	O	I-protein
aldehyde	NN	O	B-protein
dehydrogenase	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
Aldh1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
initially	NN	O	O
called	NN	O	O
Hdg-1	NN	O	B-protein
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
ectopically	NN	O	O
activated	NN	O	O
by	NN	O	O
HOX11	NN	O	B-protein
in	NN	O	O
this	NN	O	O
system	NN	O	O
.	NN	O	O

Study	NN	O	O
of	NN	O	O
Aldh1	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
spleen	NN	O	O
development	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Aldh1	NN	O	B-RNA
mRNA	NN	O	I-RNA
inversely	NN	O	O
correlated	NN	O	O
with	NN	O	O
Hox11	NN	O	B-protein
.	NN	O	O

Hox11	NN	O	B-protein
null	NN	O	O
mouse	NN	O	O
embryos	NN	O	O
have	NN	O	O
elevated	NN	O	O
Aldh1	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
spleen	NN	O	O
primordia	NN	O	O
prior	NN	O	O
to	NN	O	O
atrophy	NN	O	O
,	NN	O	O
while	NN	O	O
Aldh1	NN	O	B-protein
seems	NN	O	O
to	NN	O	O
be	NN	O	O
repressed	NN	O	O
by	NN	O	O
Hox11	NN	O	B-protein
during	NN	O	O
organogenesis	NN	O	O
of	NN	O	O
the	NN	O	O
spleens	NN	O	O
of	NN	O	O
wild-type	NN	O	O
mice	NN	O	O
.	NN	O	O

This	NN	O	O
result	NN	O	O
suggests	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
Aldh1	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
negatively	NN	O	O
regulated	NN	O	O
by	NN	O	O
Hox11	NN	O	B-protein
and	NN	O	O
that	NN	O	O
abnormal	NN	O	O
expression	NN	O	O
of	NN	O	O
Aldh1	NN	O	B-protein
in	NN	O	O
Hox11	NN	O	B-protein
null	NN	O	O
mice	NN	O	O
may	NN	O	O
cause	NN	O	O
loss	NN	O	O
of	NN	O	O
splenic	NN	O	B-cell_type
precursor	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
aberrant	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
metabolism	NN	O	O

-DOCSTART-	O

Role	NN	O	O
of	NN	O	O
the	NN	O	O
X2	NN	O	B-DNA
box	NN	O	I-DNA
in	NN	O	O
activated	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
evolutionary	NN	O	O
conserved	NN	O	O
X2	NN	O	B-DNA
box	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
HLA-DRA	NN	O	B-DNA
gene	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
human	NN	O	B-protein
major	NN	O	I-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	O
MHC	NN	O	B-protein
)	NN	O	O
in	NN	O	O
resting	NN	O	O
and	NN	O	O
activated	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NF-X2	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
contains	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1/ATF/CREB	NN	O	B-protein
families	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
X2	NN	O	B-DNA
box	NN	O	I-DNA
(	NN	O	O
5'-TGCGTCA-3	NN	O	O
'	NN	O	O
)	NN	O	O
from	NN	O	O
positions	NN	O	O
-97	NN	O	O
to	NN	O	O
-91	NN	O	O
in	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
resting	NN	O	O
Raji	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
little	NN	O	O
to	NN	O	O
no	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
X2	NN	O	B-DNA
box	NN	O	I-DNA
was	NN	O	O
observed	NN	O	O
.	NN	O	O

In	NN	O	O
sharp	NN	O	O
contrast	NN	O	O
,	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
the	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
,	NN	O	O
strong	NN	O	O
interactions	NN	O	O
between	NN	O	O
the	NN	O	O
X2	NN	O	B-DNA
box	NN	O	I-DNA
and	NN	O	O
NF-X2	NN	O	B-DNA
containing	NN	O	O
c-Fos	NN	O	B-protein
were	NN	O	O
observed	NN	O	O
.	NN	O	O

As	NN	O	O
determined	NN	O	O
by	NN	O	O
transient	NN	O	O
expression	NN	O	O
and	NN	O	O
RNA	NN	O	O
analyses	NN	O	O
,	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	O
kinase	NN	O	O
C	NN	O	O
(	NN	O	O
PKC	NN	O	O
)	NN	O	O
also	NN	O	O
increased	NN	O	O
rates	NN	O	O
of	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
wild-type	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
from	NN	O	O
a	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
bearing	NN	O	O
clustered	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
X2	NN	O	B-DNA
box	NN	O	I-DNA
.	NN	O	O

Since	NN	O	O
the	NN	O	O
co-expression	NN	O	O
with	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
c-Fos	NN	O	B-protein
abolished	NN	O	O
the	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
TPA	NN	O	O
,	NN	O	O
we	NN	O	O
conclude	NN	O	O
that	NN	O	O
activated	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
gene	NN	O	I-DNA
depends	NN	O	O
on	NN	O	O
interactions	NN	O	O
between	NN	O	O
the	NN	O	O
X2	NN	O	B-DNA
box	NN	O	I-DNA
and	NN	O	O
NF-X2	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
contains	NN	O	O
c-Fos	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Thymocytes	NN	O	O
control	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
differently	NN	O	O
from	NN	O	O
mature	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
silencer	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD4	NN	O	I-DNA
gene	NN	O	I-DNA
during	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
using	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Immunofluorescence	NN	O	O
studies	NN	O	O
on	NN	O	O
thymic	NN	O	B-cell_type
populations	NN	O	I-cell_type
of	NN	O	O
mice	NN	O	O
carrying	NN	O	O
transgenes	NN	O	O
in	NN	O	O
various	NN	O	O
combinations	NN	O	O
of	NN	O	O
these	NN	O	O
regulatory	NN	O	B-DNA
DNA	NN	O	I-DNA
elements	NN	O	I-DNA
revealed	NN	O	O
that	NN	O	O
thymocytes	NN	O	B-cell_type
control	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
different	NN	O	O
manner	NN	O	O
than	NN	O	O
mature	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
5'-positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
unit	NN	O	I-DNA
,	NN	O	O
consisting	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
already	NN	O	O
active	NN	O	O
at	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
-	NN	O	O
CD8	NN	O	B-protein
-double-negative	NN	O	O
(	NN	O	O
DN	NN	O	O
)	NN	O	O
stage	NN	O	O
of	NN	O	O
development	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
its	NN	O	O
activity	NN	O	O
becomes	NN	O	O
lower	NN	O	O
in	NN	O	O
the	NN	O	O
double-positive	NN	O	O
and	NN	O	O
a	NN	O	O
fraction	NN	O	O
of	NN	O	O
the	NN	O	O
CD4+	NN	O	B-cell_type
CD8int/-	NN	O	I-cell_type
cell	NN	O	I-cell_type
population	NN	O	I-cell_type
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
an	NN	O	O
additional	NN	O	O
enhancer	NN	O	O
,	NN	O	O
located	NN	O	O
in	NN	O	O
either	NN	O	O
the	NN	O	O
first	NN	O	B-DNA
or	NN	O	I-DNA
the	NN	O	I-DNA
third	NN	O	I-DNA
intron	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
this	NN	O	O
population	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
studied	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
is	NN	O	O
the	NN	O	O
minimal	NN	O	B-DNA
CD4	NN	O	I-DNA
silencer	NN	O	I-DNA
which	NN	O	O
inhibits	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
CD8	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
silencer	NN	O	O
is	NN	O	O
inactive	NN	O	O
in	NN	O	O
the	NN	O	O
most	NN	O	O
immature	NN	O	B-cell_type
DN	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
probably	NN	O	O
use	NN	O	O
a	NN	O	O
distinct	NN	O	O
silencer	NN	O	O
mechanism	NN	O	O
to	NN	O	O
down-regulate	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
silencer	NN	O	I-DNA
is	NN	O	O
also	NN	O	O
active	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_type
CD8int/-	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
thymus	NN	O	O
,	NN	O	O
implying	NN	O	O
that	NN	O	O
an	NN	O	O
anti-silencer	NN	O	O
may	NN	O	O
be	NN	O	O
required	NN	O	O
to	NN	O	O
resume	NN	O	O
CD4	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
this	NN	O	O
cell	NN	O	O
population	NN	O	O
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
several	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
which	NN	O	O
come	NN	O	O
into	NN	O	O
play	NN	O	O
in	NN	O	O
a	NN	O	O
developmentally	NN	O	O
coordinated	NN	O	O
manner	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
by	NN	O	O
glucocorticoids	NN	O	O
:	NN	O	O
between	NN	O	O
physiology	NN	O	O
and	NN	O	O
pharmacology	NN	O	O
]	NN	O	O

Glucocorticoids	NN	O	O
are	NN	O	O
physiological	NN	O	O
molecules	NN	O	O
that	NN	O	O
are	NN	O	O
also	NN	O	O
extensively	NN	O	O
used	NN	O	O
in	NN	O	O
clinics	NN	O	O
as	NN	O	O
anti-inflammatory	NN	O	O
,	NN	O	O
immunosuppressive	NN	O	O
or	NN	O	O
anti-tumoral	NN	O	O
agents	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
can	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
on	NN	O	O
normal	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
physiology	NN	O	O
of	NN	O	O
thymic	NN	O	O
selection	NN	O	O
.	NN	O	O

In	NN	O	O
clinics	NN	O	O
these	NN	O	O
molecules	NN	O	O
are	NN	O	O
also	NN	O	O
used	NN	O	O
for	NN	O	O
their	NN	O	O
potencies	NN	O	O
in	NN	O	O
inducing	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
malignant	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Glucocorticoids	NN	O	O
are	NN	O	O
mediating	NN	O	O
their	NN	O	O
effects	NN	O	O
after	NN	O	O
binding	NN	O	O
to	NN	O	O
an	NN	O	O
intracellular	NN	O	B-protein
receptor	NN	O	I-protein
belonging	NN	O	O
to	NN	O	O
the	NN	O	O
steroid	NN	O	B-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
:	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Once	NN	O	O
activated	NN	O	O
,	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
,	NN	O	O
can	NN	O	O
mediate	NN	O	O
his	NN	O	O
effects	NN	O	O
through	NN	O	O
direct	NN	O	O
binding	NN	O	O
on	NN	O	O
the	NN	O	O
DNA	NN	O	O
or	NN	O	O
via	NN	O	O
protein/protein	NN	O	O
interactions	NN	O	O
with	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Depending	NN	O	O
on	NN	O	O
the	NN	O	O
type	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
fall	NN	O	O
roughly	NN	O	O
in	NN	O	O
two	NN	O	O
categories	NN	O	O
:	NN	O	O
induction	NN	O	O
of	NN	O	O
``	NN	O	O
death	NN	O	O
genes	NN	O	O
''	NN	O	O
by	NN	O	O
the	NN	O	O
activated	NN	O	O
GR	NN	O	B-protein
(	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
c-jun	NN	O	B-protein
)	NN	O	O
or	NN	O	O
repression	NN	O	O
of	NN	O	O
survival	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
c-Myc	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
thymic	NN	O	O
selection	NN	O	O
the	NN	O	O
mechanism	NN	O	O
is	NN	O	O
more	NN	O	O
subtle	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
mutual	NN	O	O
repression	NN	O	O
of	NN	O	O
Nur77	NN	O	B-protein
and	NN	O	O
GR	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Direct	NN	O	O
suppression	NN	O	O
of	NN	O	O
Stat1	NN	O	B-protein
function	NN	O	O
during	NN	O	O
adenoviral	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
action	NN	O	O
of	NN	O	O
adenoviral	NN	O	B-protein
E1A	NN	O	I-protein
oncoprotein	NN	O	I-protein
on	NN	O	O
host	NN	O	B-DNA
immune-response	NN	O	I-DNA
genes	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
attributed	NN	O	O
to	NN	O	O
interaction	NN	O	O
with	NN	O	O
p300/CBP-type	NN	O	B-protein
transcriptional	NN	O	I-protein
coactivators	NN	O	I-protein
in	NN	O	O
competition	NN	O	O
with	NN	O	O
endogenous	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	I-protein
STAT	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
E1A	NN	O	B-protein
that	NN	O	O
no	NN	O	O
longer	NN	O	O
bind	NN	O	O
p300/CBP	NN	O	B-protein
can	NN	O	O
still	NN	O	O
interact	NN	O	O
directly	NN	O	O
with	NN	O	O
Stat1	NN	O	B-protein
(	NN	O	O
via	NN	O	O
E1A	NN	O	B-protein
N-terminal	NN	O	I-protein
and	NN	O	I-protein
Stat1	NN	O	I-protein
C-terminal	NN	O	I-protein
residues	NN	O	I-protein
)	NN	O	O
and	NN	O	O
block	NN	O	O
IFNgamma-driven	NN	O	O
,	NN	O	O
Stat1	NN	O	B-protein
-dependent	NN	O	O
gene	NN	O	O
activation	NN	O	O
and	NN	O	O
consequent	NN	O	O
function	NN	O	O
during	NN	O	O
early-phase	NN	O	O
infection	NN	O	O
in	NN	O	O
the	NN	O	O
natural	NN	O	B-cell_type
host	NN	O	I-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
results	NN	O	O
provide	NN	O	O
a	NN	O	O
distinct	NN	O	O
and	NN	O	O
more	NN	O	O
specific	NN	O	O
mechanism	NN	O	O
for	NN	O	O
E1A	NN	O	B-protein
-mediated	NN	O	O
immune	NN	O	O
suppression	NN	O	O
and	NN	O	O
an	NN	O	O
alternative	NN	O	O
model	NN	O	O
of	NN	O	O
IFNgamma	NN	O	B-protein
-driven	NN	O	O
enhanceosome	NN	O	O
formation	NN	O	O
that	NN	O	O
may	NN	O	O
allow	NN	O	O
for	NN	O	O
other	NN	O	O
adaptors	NN	O	O
(	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
p300/CBP	NN	O	B-protein
)	NN	O	O
to	NN	O	O
link	NN	O	O
Stat1	NN	O	B-protein
to	NN	O	O
the	NN	O	O
basal	NN	O	B-protein
transcription	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Effect	NN	O	O
of	NN	O	O
environmental	NN	O	O
estrogens	NN	O	O
on	NN	O	O
IL-1beta	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
a	NN	O	O
macrophage	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Environmental	NN	O	O
estrogens	NN	O	O
or	NN	O	O
estrogen	NN	O	O
disrupters	NN	O	O
have	NN	O	O
recently	NN	O	O
received	NN	O	O
a	NN	O	O
great	NN	O	O
deal	NN	O	O
of	NN	O	O
attention	NN	O	O
because	NN	O	O
of	NN	O	O
their	NN	O	O
potential	NN	O	O
health	NN	O	O
impact	NN	O	O
on	NN	O	O
reproductive	NN	O	O
tissues	NN	O	O
.	NN	O	O

Few	NN	O	O
,	NN	O	O
if	NN	O	O
any	NN	O	O
,	NN	O	O
studies	NN	O	O
have	NN	O	O
been	NN	O	O
made	NN	O	O
on	NN	O	O
the	NN	O	O
impact	NN	O	O
of	NN	O	O
these	NN	O	O
compounds	NN	O	O
on	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

We	NN	O	O
sought	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
various	NN	O	O
environmental	NN	O	O
estrogens	NN	O	O
on	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-1beta	NN	O	B-DNA
(	NN	O	I-DNA
IL-1beta	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
model	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
hER	NN	O	B-cell_line
+	NN	O	I-cell_line
IL-1beta-CAT+	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
cell	NN	O	O
line	NN	O	O
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	B-protein
estrogen	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
an	NN	O	O
IL-1beta	NN	O	B-DNA
promoter	NN	O	I-DNA
construct	NN	O	O
fused	NN	O	O
to	NN	O	O
the	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
allows	NN	O	O
us	NN	O	O
to	NN	O	O
monitor	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
estrogenic	NN	O	O
compounds	NN	O	O
on	NN	O	O
IL-1beta	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

17beta-estradiol	NN	O	O
(	NN	O	O
E2	NN	O	O
)	NN	O	O
markedly	NN	O	O
enhanced	NN	O	O
lipopolysaccharide	NN	O	O
-	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
induced	NN	O	O
IL-1beta	NN	O	B-DNA
promoter	NN	O	I-DNA
-driven	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

The	NN	O	O
mycotoxins	NN	O	O
alpha-zearalenol	NN	O	O
and	NN	O	O
zearalenone	NN	O	O
both	NN	O	O
exhibited	NN	O	O
full	NN	O	O
agonist	NN	O	O
activity	NN	O	O
,	NN	O	O
but	NN	O	O
at	NN	O	O
lower	NN	O	O
potencies	NN	O	O
,	NN	O	O
with	NN	O	O
EC50	NN	O	O
values	NN	O	O
of	NN	O	O
1.8	NN	O	O
and	NN	O	O
54	NN	O	O
nM	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
compared	NN	O	O
with	NN	O	O
E2	NN	O	O
at	NN	O	O
0.5	NN	O	O
nM	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
genistein	NN	O	O
was	NN	O	O
a	NN	O	O
very	NN	O	O
low-potency	NN	O	O
agonist	NN	O	O
,	NN	O	O
having	NN	O	O
an	NN	O	O
EC50	NN	O	O
of	NN	O	O
1.5	NN	O	O
microM	NN	O	O
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
the	NN	O	O
E2	NN	O	O
response	NN	O	O
,	NN	O	O
the	NN	O	O
slope	NN	O	O
factors	NN	O	O
for	NN	O	O
alpha-zearalenol	NN	O	O
,	NN	O	O
zearalenone	NN	O	O
,	NN	O	O
and	NN	O	O
genistein	NN	O	O
were	NN	O	O
close	NN	O	O
to	NN	O	O
3.0	NN	O	O
,	NN	O	O
suggesting	NN	O	O
positive	NN	O	O
cooperativity	NN	O	O
in	NN	O	O
the	NN	O	O
estrogenic	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
mycotoxins	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
estrogen	NN	O	O
receptor	NN	O	O
,	NN	O	O
since	NN	O	O
both	NN	O	O
the	NN	O	O
antiestrogens	NN	O	O
H1285	NN	O	O
and	NN	O	O
ICI	NN	O	O
182	NN	O	O
,	NN	O	O
780	NN	O	O
effectively	NN	O	O
inhibited	NN	O	O
their	NN	O	O
agonist	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Representative	NN	O	O
environmental	NN	O	O
estrogenic	NN	O	O
compounds	NN	O	O
both	NN	O	O
from	NN	O	O
plant	NN	O	O
and	NN	O	O
industrial	NN	O	O
sources	NN	O	O
were	NN	O	O
also	NN	O	O
tested	NN	O	O
.	NN	O	O

Unlike	NN	O	O
the	NN	O	O
mycoestrogens	NN	O	O
,	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
compounds	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
genistein	NN	O	O
,	NN	O	O
synergized	NN	O	O
with	NN	O	O
LPS	NN	O	O
to	NN	O	O
enhance	NN	O	O
IL-1beta	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

When	NN	O	O
tested	NN	O	O
for	NN	O	O
antiestrogenic	NN	O	O
activity	NN	O	O
,	NN	O	O
the	NN	O	O
industrial	NN	O	O
compound	NN	O	O
4-octylphenol	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
antagonize	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
E2	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
response	NN	O	O
was	NN	O	O
three	NN	O	O
orders	NN	O	O
of	NN	O	O
magnitude	NN	O	O
less	NN	O	O
potent	NN	O	O
than	NN	O	O
H	NN	O	O
1285	NN	O	O
.	NN	O	O

Naringenin	NN	O	O
,	NN	O	O
a	NN	O	O
plant	NN	O	O
flavonoid	NN	O	O
,	NN	O	O
showed	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
ability	NN	O	O
to	NN	O	O
antagonize	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
E2	NN	O	O
.	NN	O	O

Overall	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
some	NN	O	O
environmental	NN	O	O
estrogens	NN	O	O
that	NN	O	O
display	NN	O	O
agonist	NN	O	O
activity	NN	O	O
in	NN	O	O
reproductive	NN	O	O
tissue	NN	O	O
also	NN	O	O
have	NN	O	O
an	NN	O	O
effect	NN	O	O
on	NN	O	O
IL-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
hemopoietic-derived	NN	O	O
tissue	NN	O	O
.	NN	O	O

-DOCSTART-	O

Isolation	NN	O	O
and	NN	O	O
utilization	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
to	NN	O	O
assay	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
primary	NN	O	O
immune	NN	O	O
response	NN	O	O
to	NN	O	O
varicella-zoster	NN	O	O
virus	NN	O	O
peptides	NN	O	O
.	NN	O	O

A	NN	O	O
human	NN	O	O
dendritic	NN	O	O
cell-based	NN	O	O
assay	NN	O	O
used	NN	O	O
to	NN	O	O
monitor	NN	O	O
a	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
response	NN	O	O
to	NN	O	O
viral	NN	O	O
peptides	NN	O	O
in	NN	O	O
vitro	NN	O	O
is	NN	O	O
described	NN	O	O
.	NN	O	O

Dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
autologous	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
isolated	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
by	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
density-gradient	NN	O	O
centrifugations	NN	O	O
or	NN	O	O
magnetic	NN	O	O
bead	NN	O	O
separations	NN	O	O
(	NN	O	O
or	NN	O	O
both	NN	O	O
)	NN	O	O
.	NN	O	O

Peptides	NN	O	O
corresponding	NN	O	O
to	NN	O	O
residues	NN	O	O
of	NN	O	O
the	NN	O	O
immediate	NN	O	B-protein
early	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
IE62	NN	O	B-protein
,	NN	O	O
of	NN	O	O
varicella-zoster	NN	O	O
virus	NN	O	O
(	NN	O	O
VZV	NN	O	O
)	NN	O	O
were	NN	O	O
used	NN	O	O
as	NN	O	O
stimulating	NN	O	B-protein
antigens	NN	O	I-protein
,	NN	O	O
and	NN	O	O
persons	NN	O	O
with	NN	O	O
no	NN	O	O
history	NN	O	O
of	NN	O	O
varicella	NN	O	O
and	NN	O	O
no	NN	O	O
humoral	NN	O	O
or	NN	O	O
cellular	NN	O	O
immunity	NN	O	O
to	NN	O	O
VZV	NN	O	O
served	NN	O	O
as	NN	O	O
naive	NN	O	O
donors	NN	O	O
for	NN	O	O
the	NN	O	O
assays	NN	O	O
.	NN	O	O

Three	NN	O	O
VZV-susceptible	NN	O	O
donors	NN	O	O
were	NN	O	O
tested	NN	O	O
,	NN	O	O
and	NN	O	O
all	NN	O	O
demonstrated	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
response	NN	O	O
to	NN	O	O
multiple	NN	O	O
VZV	NN	O	O
peptides	NN	O	O
.	NN	O	O

This	NN	O	O
assay	NN	O	O
has	NN	O	O
potential	NN	O	O
as	NN	O	O
a	NN	O	O
screen	NN	O	O
to	NN	O	O
establish	NN	O	O
the	NN	O	O
immunogenicity	NN	O	O
of	NN	O	O
viral	NN	O	O
antigens	NN	O	O
in	NN	O	O
vitro	NN	O	O
using	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
naive	NN	O	O
donors	NN	O	O
.	NN	O	O

-DOCSTART-	O

Xenogeneic	NN	O	O
human	NN	O	O
serum	NN	O	O
promotes	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
porcine	NN	O	O
endothelium	NN	O	O
under	NN	O	O
flow	NN	O	O
conditions	NN	O	O
,	NN	O	O
possibly	NN	O	O
through	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Endothelial	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
leukocyte	NN	O	O
infiltration	NN	O	O
are	NN	O	O
a	NN	O	O
consistent	NN	O	O
feature	NN	O	O
of	NN	O	O
discordant	NN	O	O
xenograft	NN	O	O
rejection	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
evaluated	NN	O	O
whether	NN	O	O
xenogeneic	NN	O	O
serum	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
source	NN	O	O
of	NN	O	O
xenoreactive	NN	O	B-protein
natural	NN	O	I-protein
antibodies	NN	O	I-protein
and	NN	O	O
complement	NN	O	O
,	NN	O	O
induced	NN	O	O
endothelial	NN	O	O
cell	NN	O	O
activation	NN	O	O
with	NN	O	O
consequent	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
under	NN	O	O
flow	NN	O	O
conditions	NN	O	O
.	NN	O	O

Porcine	NN	O	B-cell_line
aortic	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
PAEC	NN	O	B-cell_line
)	NN	O	O
were	NN	O	O
incubated	NN	O	O
for	NN	O	O
1	NN	O	O
hr	NN	O	O
30	NN	O	O
min	NN	O	O
or	NN	O	O
5	NN	O	O
hr	NN	O	O
with	NN	O	O
10	NN	O	O
%	NN	O	O
homologous	NN	O	O
porcine	NN	O	O
serum	NN	O	O
(	NN	O	O
control	NN	O	O
)	NN	O	O
or	NN	O	O
10	NN	O	O
%	NN	O	O
xenogeneic	NN	O	O
human	NN	O	O
serum	NN	O	O
and	NN	O	O
then	NN	O	O
perfused	NN	O	O
with	NN	O	O
total	NN	O	O
human	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
a	NN	O	O
parallel	NN	O	O
plate	NN	O	O
flow	NN	O	O
chamber	NN	O	O
under	NN	O	O
laminar	NN	O	O
flow	NN	O	O
(	NN	O	O
1.5	NN	O	O
dynes/cm2	NN	O	O
)	NN	O	O
.	NN	O	O

Adherent	NN	O	O
cells	NN	O	O
were	NN	O	O
counted	NN	O	O
by	NN	O	O
digital	NN	O	O
image	NN	O	O
analysis	NN	O	O
.	NN	O	O

Xenogeneic	NN	O	O
human	NN	O	O
serum	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
increased	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
adherent	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
as	NN	O	O
compared	NN	O	O
with	NN	O	O
porcine	NN	O	O
serum	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
adhesive	NN	O	O
response	NN	O	O
was	NN	O	O
elicited	NN	O	O
by	NN	O	O
TNF	NN	O	O
alpha	NN	O	O
(	NN	O	O
100	NN	O	O
U/ml	NN	O	O
)	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
potent	NN	O	O
inducers	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cell	NN	O	I-cell_type
adhesive	NN	O	O
properties	NN	O	O
,	NN	O	O
here	NN	O	O
used	NN	O	O
as	NN	O	O
positive	NN	O	O
control	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
elucidate	NN	O	O
possible	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
endothelial	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
by	NN	O	O
xenogeneic	NN	O	O
serum	NN	O	O
,	NN	O	O
we	NN	O	O
focussed	NN	O	O
on	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
central	NN	O	O
regulator	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
different	NN	O	O
genes	NN	O	O
,	NN	O	O
including	NN	O	O
adhesive	NN	O	B-protein
molecules	NN	O	I-protein
and	NN	O	O
chemoattractants	NN	O	B-protein
.	NN	O	O

By	NN	O	O
confocal	NN	O	O
fluorescence	NN	O	O
microscopy	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
a	NN	O	O
positive	NN	O	O
staining	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
p65	NN	O	B-protein
subunit	NN	O	I-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
PAEC	NN	O	B-cell_line
exposed	NN	O	O
for	NN	O	O
1	NN	O	O
hr	NN	O	O
30	NN	O	O
min	NN	O	O
to	NN	O	O
human	NN	O	O
serum	NN	O	O
,	NN	O	O
which	NN	O	O
indicated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
this	NN	O	O
setting	NN	O	O
.	NN	O	O

At	NN	O	O
variance	NN	O	O
,	NN	O	O
in	NN	O	O
PAEC	NN	O	B-cell_line
incubated	NN	O	O
with	NN	O	O
the	NN	O	O
homologous	NN	O	O
serum	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
strictly	NN	O	O
localized	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
PAEC	NN	O	B-cell_line
exposed	NN	O	O
to	NN	O	O
xenogeneic	NN	O	O
serum	NN	O	O
with	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
inhibitors	NN	O	O
pyrrolidinedithiocarbamate	NN	O	O
(	NN	O	O
PDTC	NN	O	O
,	NN	O	O
25	NN	O	O
microM	NN	O	O
)	NN	O	O
and	NN	O	O
tosyl-phechloromethylketone	NN	O	O
(	NN	O	O
TPCK	NN	O	O
,	NN	O	O
25	NN	O	O
microM	NN	O	O
)	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
reduced	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
in	NN	O	O
respect	NN	O	O
to	NN	O	O
PAEC	NN	O	B-cell_line
treated	NN	O	O
with	NN	O	O
human	NN	O	O
serum	NN	O	O
alone	NN	O	O
.	NN	O	O

Findings	NN	O	O
that	NN	O	O
xenogeneic	NN	O	O
serum	NN	O	O
promotes	NN	O	O
leukocyte-endothelium	NN	O	O
interaction	NN	O	O
possibly	NN	O	O
through	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
might	NN	O	O
be	NN	O	O
relevant	NN	O	O
for	NN	O	O
designing	NN	O	O
future	NN	O	O
therapeutic	NN	O	O
strategies	NN	O	O
aimed	NN	O	O
at	NN	O	O
prolonging	NN	O	O
xenograft	NN	O	O
survival	NN	O	O
.	NN	O	O

-DOCSTART-	O

Anaphylatoxins	NN	O	B-protein
C5a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
induce	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
anaphylatoxins	NN	O	B-protein
C5a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	O
production	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
the	NN	O	O
capability	NN	O	O
of	NN	O	O
C5a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
transcription	NN	O	O
factor	NN	O	O
activation	NN	O	O
was	NN	O	O
examined	NN	O	O
.	NN	O	O

C5a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
resulted	NN	O	O
in	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
with	NN	O	O
specificity	NN	O	O
to	NN	O	O
the	NN	O	O
kappaB	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
p50	NN	O	B-protein
and	NN	O	I-protein
p65	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
constituents	NN	O	O
of	NN	O	O
the	NN	O	O
prototypic	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
,	NN	O	O
were	NN	O	O
identified	NN	O	O
as	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-protein	NN	O	O
complexes	NN	O	O
by	NN	O	O
anti-peptide	NN	O	B-protein
antibodies	NN	O	I-protein
in	NN	O	O
gel	NN	O	O
supershift	NN	O	O
assays	NN	O	O
.	NN	O	O

C5a	NN	O	B-protein
induced	NN	O	O
kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
detected	NN	O	O
15	NN	O	O
min	NN	O	O
after	NN	O	O
agonist	NN	O	O
stimulation	NN	O	O
,	NN	O	O
peaked	NN	O	O
at	NN	O	O
30-40	NN	O	O
min	NN	O	O
,	NN	O	O
and	NN	O	O
remained	NN	O	O
detectable	NN	O	O
at	NN	O	O
2	NN	O	O
h	NN	O	O
.	NN	O	O

Binding	NN	O	O
to	NN	O	O
kappaB	NN	O	B-DNA
sequence	NN	O	I-DNA
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
initial	NN	O	O
decrease	NN	O	O
and	NN	O	O
subsequent	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
IkappaBalpha	NN	O	B-protein
levels	NN	O	O
,	NN	O	O
as	NN	O	O
detected	NN	O	O
by	NN	O	O
Western	NN	O	O
blotting	NN	O	O
using	NN	O	O
an	NN	O	O
anti-IkappaBalpha	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

Pertussis	NN	O	O
toxin	NN	O	O
treatment	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
kappaB	NN	O	B-protein
binding	NN	O	O
activities	NN	O	O
induced	NN	O	O
by	NN	O	O
both	NN	O	O
C5a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
cholera	NN	O	B-protein
toxin	NN	O	I-protein
displayed	NN	O	O
no	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
.	NN	O	O

Neither	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
toxins	NN	O	O
affected	NN	O	O
kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
induced	NN	O	O
by	NN	O	O
TNFalpha	NN	O	B-protein
in	NN	O	O
the	NN	O	O
same	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
imply	NN	O	O
a	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
anaphylatoxins	NN	O	B-protein
C5a	NN	O	B-protein
and	NN	O	O
C3a	NN	O	B-protein
in	NN	O	O
regulating	NN	O	O
leukocytes	NN	O	O
gene	NN	O	O
expression	NN	O	O
through	NN	O	O
G	NN	O	O
protein-coupled	NN	O	O
transcription	NN	O	O
factor	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Thrombopoietin	NN	O	B-protein
and	NN	O	O
its	NN	O	O
receptor	NN	O	O
.	NN	O	O

Thrombopoietin	NN	O	B-protein
(	NN	O	O
TPO	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
primary	NN	O	O
physiological	NN	O	O
regulator	NN	O	O
of	NN	O	O
platelet	NN	O	O
production	NN	O	O
,	NN	O	O
was	NN	O	O
initially	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
lineage-specific	NN	O	O
factor	NN	O	O
acting	NN	O	O
predominantly	NN	O	O
on	NN	O	O
megakaryocytopoiesis	NN	O	O
.	NN	O	O

Detailed	NN	O	O
studies	NN	O	O
establish	NN	O	O
that	NN	O	O
this	NN	O	O
cytokine	NN	O	O
mediates	NN	O	O
biological	NN	O	O
effects	NN	O	O
on	NN	O	O
a	NN	O	O
broad	NN	O	O
spectrum	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
including	NN	O	O
stem	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

TPO	NN	O	B-protein
is	NN	O	O
a	NN	O	O
hormone	NN	O	O
constitutively	NN	O	O
produced	NN	O	O
mainly	NN	O	O
by	NN	O	O
the	NN	O	O
liver	NN	O	O
and	NN	O	O
kidney	NN	O	O
.	NN	O	O

Plasma	NN	O	O
TPO	NN	O	B-protein
levels	NN	O	O
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
platelet	NN	O	O
and	NN	O	O
megakaryocyte	NN	O	O
mass	NN	O	O
through	NN	O	O
Mpl	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
,	NN	O	O
internalization	NN	O	O
and	NN	O	O
degradation	NN	O	O
.	NN	O	O

The	NN	O	O
Mpl	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
hematopoietin	NN	O	B-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
lacking	NN	O	O
intrinsic	NN	O	O
kinase	NN	O	O
activity	NN	O	O
.	NN	O	O

Upon	NN	O	O
ligand-induced	NN	O	O
Mpl	NN	O	O
homodimerization	NN	O	O
,	NN	O	O
the	NN	O	O
major	NN	O	O
signaling	NN	O	O
events	NN	O	O
for	NN	O	O
proliferation	NN	O	O
are	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
JAK2	NN	O	B-protein
/STAT5	NN	O	B-protein
pathway	NN	O	O
,	NN	O	O
while	NN	O	O
differentiation	NN	O	O
might	NN	O	O
occur	NN	O	O
through	NN	O	O
a	NN	O	O
prolonged	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
MAPK	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

Preclinical	NN	O	O
and	NN	O	O
clinical	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
potential	NN	O	O
use	NN	O	O
of	NN	O	O
TPO	NN	O	B-protein
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
contexts	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
is	NN	O	O
too	NN	O	O
early	NN	O	O
to	NN	O	O
evaluate	NN	O	O
its	NN	O	O
benefit	NN	O	O
in	NN	O	O
reducing	NN	O	O
platelet	NN	O	O
transfusion	NN	O	O
.	NN	O	O

-DOCSTART-	O

TAL1	NN	O	B-protein
and	NN	O	O
LIM-only	NN	O	B-protein
proteins	NN	O	I-protein
synergistically	NN	O	O
induce	NN	O	O
retinaldehyde	NN	O	B-protein
dehydrogenase	NN	O	I-protein
2	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
T-cell	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
by	NN	O	O
acting	NN	O	O
as	NN	O	O
cofactors	NN	O	O
for	NN	O	O
GATA3	NN	O	B-protein
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
TAL1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
LIM-only	NN	O	B-DNA
protein	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
LMO	NN	O	B-DNA
)	NN	O	O
are	NN	O	O
regularly	NN	O	O
coactivated	NN	O	O
in	NN	O	O
T-cell	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
T-ALL	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
relate	NN	O	O
to	NN	O	O
the	NN	O	O
findings	NN	O	O
that	NN	O	O
TAL1	NN	O	B-protein
and	NN	O	O
LMO	NN	O	B-protein
are	NN	O	O
highly	NN	O	O
synergistic	NN	O	O
in	NN	O	O
T-cell	NN	O	O
tumorigenesis	NN	O	O
in	NN	O	O
double-transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
functional	NN	O	O
synergy	NN	O	O
between	NN	O	O
TAL1	NN	O	B-protein
and	NN	O	O
LMO	NN	O	B-protein
in	NN	O	O
tumorigenesis	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
,	NN	O	O
we	NN	O	O
tried	NN	O	O
to	NN	O	O
identify	NN	O	O
downstream	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
regulated	NN	O	O
by	NN	O	O
TAL1	NN	O	B-protein
and	NN	O	O
LMO	NN	O	B-protein
by	NN	O	O
a	NN	O	O
subtractive	NN	O	O
PCR	NN	O	O
method	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
isolated	NN	O	O
genes	NN	O	O
,	NN	O	O
that	NN	O	O
for	NN	O	O
retinaldehyde	NN	O	B-protein
dehydrogenase	NN	O	I-protein
2	NN	O	I-protein
(	NN	O	O
RALDH2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
was	NN	O	O
regularly	NN	O	O
expressed	NN	O	O
in	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
T-ALL	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
coexpressed	NN	O	O
TAL1	NN	O	B-protein
and	NN	O	O
LMO	NN	O	B-protein
.	NN	O	O

Exogenously	NN	O	O
transfected	NN	O	O
TAL1	NN	O	B-protein
and	NN	O	O
LMO	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
either	NN	O	O
alone	NN	O	O
,	NN	O	O
induced	NN	O	O
RALDH2	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
a	NN	O	O
T-ALL	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
HPB-ALL	NN	O	B-cell_line
,	NN	O	O
not	NN	O	O
expressing	NN	O	O
endogeneous	NN	O	O
TAL1	NN	O	B-protein
or	NN	O	O
LMO	NN	O	B-protein
.	NN	O	O

The	NN	O	O
RALDH2	NN	O	B-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
T-ALL	NN	O	O
were	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
mostly	NN	O	O
initiated	NN	O	O
within	NN	O	O
the	NN	O	O
second	NN	O	B-RNA
intron	NN	O	I-RNA
.	NN	O	O

Promoter	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
a	NN	O	O
GATA	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
cryptic	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
second	NN	O	B-DNA
intron	NN	O	I-DNA
was	NN	O	O
essential	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
the	NN	O	O
TAL1-	NN	O	O
and	NN	O	O
LMO-dependent	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
GATA3	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
forced	NN	O	O
expression	NN	O	O
of	NN	O	O
GATA3	NN	O	B-protein
potentiated	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
RALDH2	NN	O	B-protein
by	NN	O	O
TAL1	NN	O	B-protein
and	NN	O	O
LMO	NN	O	B-protein
,	NN	O	O
and	NN	O	O
these	NN	O	O
three	NN	O	O
factors	NN	O	O
formed	NN	O	O
a	NN	O	O
complex	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
a	NN	O	O
TAL1	NN	O	B-protein
mutant	NN	O	O
not	NN	O	O
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
also	NN	O	O
activated	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
RALDH2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
LMO	NN	O	B-protein
and	NN	O	O
GATA3	NN	O	B-protein
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
the	NN	O	O
RALDH2	NN	O	B-DNA
gene	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
first	NN	O	O
example	NN	O	O
of	NN	O	O
direct	NN	O	O
transcriptional	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
regulated	NN	O	O
by	NN	O	O
TAL1	NN	O	B-protein
and	NN	O	O
LMO	NN	O	B-protein
in	NN	O	O
T-ALL	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
case	NN	O	O
,	NN	O	O
TAL1	NN	O	B-protein
and	NN	O	O
LMO	NN	O	B-protein
act	NN	O	O
as	NN	O	O
cofactors	NN	O	O
for	NN	O	O
GATA3	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
RALDH2	NN	O	B-protein

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
relation	NN	O	O
to	NN	O	O
age	NN	O	O
and	NN	O	O
to	NN	O	O
sport	NN	O	O
activity	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
are	NN	O	O
ubiquitous	NN	O	O
molecules	NN	O	O
and	NN	O	O
are	NN	O	O
present	NN	O	O
also	NN	O	O
in	NN	O	O
the	NN	O	O
hippocampus	NN	O	O
and	NN	O	O
in	NN	O	O
several	NN	O	O
other	NN	O	O
nervous	NN	O	O
and	NN	O	O
immune	NN	O	O
tissues	NN	O	O
.	NN	O	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
are	NN	O	O
a	NN	O	O
good	NN	O	O
model	NN	O	O
for	NN	O	O
studies	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
humans	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
maintaining	NN	O	O
cellular	NN	O	O
and	NN	O	O
humoral	NN	O	O
homeostasis	NN	O	O
and	NN	O	O
are	NN	O	O
key	NN	O	O
mediators	NN	O	O
of	NN	O	O
neuroendocrine-immune	NN	O	O
regulatory	NN	O	O
interactions	NN	O	O
.	NN	O	O

The	NN	O	O
increase	NN	O	O
of	NN	O	O
cortisol	NN	O	O
is	NN	O	O
immunosuppressive	NN	O	O
and	NN	O	O
reduces	NN	O	O
GR	NN	O	B-protein
concentration	NN	O	O
both	NN	O	O
in	NN	O	O
nervous	NN	O	O
and	NN	O	O
immune	NN	O	O
systems	NN	O	O
.	NN	O	O

Variation	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
healthy	NN	O	O
aged	NN	O	O
subjects	NN	O	O
and	NN	O	O
athletes	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
.	NN	O	O

Prompted	NN	O	O
by	NN	O	O
these	NN	O	O
results	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
in	NN	O	O
man	NN	O	O
a	NN	O	O
possible	NN	O	O
relationship	NN	O	O
between	NN	O	O
GR	NN	O	B-protein
binding	NN	O	O
capacity	NN	O	O
in	NN	O	O
the	NN	O	O
PBMCs	NN	O	B-cell_type
and	NN	O	O
age	NN	O	O
,	NN	O	O
in	NN	O	O
relation	NN	O	O
also	NN	O	O
to	NN	O	O
plasma	NN	O	O
testosterone	NN	O	O
and	NN	O	O
cortisol	NN	O	O
.	NN	O	O

The	NN	O	O
same	NN	O	O
parameters	NN	O	O
have	NN	O	O
been	NN	O	O
examined	NN	O	O
in	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
soccer	NN	O	O
players	NN	O	O
for	NN	O	O
comparison	NN	O	O
with	NN	O	O
the	NN	O	O
sedentary	NN	O	O
group	NN	O	O
.	NN	O	O

GR	NN	O	B-protein
binding	NN	O	O
capacity	NN	O	O
was	NN	O	O
higher	NN	O	O
in	NN	O	O
younger	NN	O	O
subjects	NN	O	O
than	NN	O	O
in	NN	O	O
older	NN	O	O
ones	NN	O	O
,	NN	O	O
and	NN	O	O
lower	NN	O	O
in	NN	O	O
the	NN	O	O
group	NN	O	O
of	NN	O	O
athletes	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
younger	NN	O	O
and	NN	O	O
older	NN	O	O
sedentary	NN	O	O
subjects	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
sedentary	NN	O	O
group	NN	O	O
a	NN	O	O
negative	NN	O	O
correlation	NN	O	O
was	NN	O	O
present	NN	O	O
between	NN	O	O
GR	NN	O	B-protein
binding	NN	O	O
capacity	NN	O	O
and	NN	O	O
age	NN	O	O
.	NN	O	O

Plasma	NN	O	O
cortisol	NN	O	O
was	NN	O	O
higher	NN	O	O
and	NN	O	O
testosterone	NN	O	O
lower	NN	O	O
in	NN	O	O
the	NN	O	O
athletes	NN	O	O
;	NN	O	O
they	NN	O	O
were	NN	O	O
negatively	NN	O	O
correlated	NN	O	O
in	NN	O	O
athletes	NN	O	O
and	NN	O	O
positively	NN	O	O
correlated	NN	O	O
in	NN	O	O
the	NN	O	O
sedentary	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
for	NN	O	O
athletes	NN	O	O
agree	NN	O	O
with	NN	O	O
their	NN	O	O
lower	NN	O	O
anabolic/catabolic	NN	O	O
balance	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
reduced	NN	O	O
GR	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
age	NN	O	O
and	NN	O	O
sport	NN	O	O
activity	NN	O	O
could	NN	O	O
involve	NN	O	O
a	NN	O	O
loss	NN	O	O
or	NN	O	O
an	NN	O	O
involution	NN	O	O
of	NN	O	O
receptor	NN	O	O
synthesis	NN	O	O
.	NN	O	O

However	NN	O	O
other	NN	O	O
possibilities	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
altered	NN	O	O
distribution	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
subpopulations	NN	O	O
with	NN	O	O
different	NN	O	O
receptor	NN	O	O
concentrations	NN	O	O
and	NN	O	O
with	NN	O	O
different	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
,	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
excluded	NN	O	O
.	NN	O	O

Several	NN	O	O
neuroendocrine-immune	NN	O	O
interactions	NN	O	O
could	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
reduced	NN	O	O
GR	NN	O	B-protein
levels	NN	O	O
with	NN	O	O
age	NN	O	O
and	NN	O	O
sport	NN	O	O
activity	NN	O	O
in	NN	O	O
man	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glycation-dependent	NN	O	O
,	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species-mediated	NN	O	O
suppression	NN	O	O
of	NN	O	O
the	NN	O	O
insulin	NN	O	O
gene	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
HIT	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Prolonged	NN	O	O
poor	NN	O	O
glycemic	NN	O	O
control	NN	O	O
in	NN	O	O
non-insulin-dependent	NN	O	O
diabetes	NN	O	O
mellitus	NN	O	O
patients	NN	O	O
often	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
decline	NN	O	O
in	NN	O	O
insulin	NN	O	O
secretion	NN	O	O
from	NN	O	O
pancreatic	NN	O	B-cell_type
beta	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
insulin	NN	O	O
content	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
step	NN	O	O
toward	NN	O	O
elucidating	NN	O	O
the	NN	O	O
pathophysiological	NN	O	O
background	NN	O	O
of	NN	O	O
the	NN	O	O
so-called	NN	O	O
glucose	NN	O	O
toxicity	NN	O	O
to	NN	O	O
pancreatic	NN	O	B-cell_type
beta	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
induced	NN	O	O
glycation	NN	O	O
in	NN	O	O
HIT-T15	NN	O	B-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
a	NN	O	O
sugar	NN	O	O
with	NN	O	O
strong	NN	O	O
deoxidizing	NN	O	O
activity	NN	O	O
,	NN	O	O
D-ribose	NN	O	O
,	NN	O	O
and	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
on	NN	O	O
insulin	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
analyses	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
insulin	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
more	NN	O	O
sensitive	NN	O	O
to	NN	O	O
glycation	NN	O	O
than	NN	O	O
the	NN	O	O
control	NN	O	O
beta-actin	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
;	NN	O	O
approximately	NN	O	O
50	NN	O	O
and	NN	O	O
80	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
insulin	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
was	NN	O	O
lost	NN	O	O
when	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
kept	NN	O	O
for	NN	O	O
3	NN	O	O
d	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
40	NN	O	O
and	NN	O	O
60	NN	O	O
mM	NN	O	O
D-ribose	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

In	NN	O	O
agreement	NN	O	O
with	NN	O	O
this	NN	O	O
,	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
insulin	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
insulin	NN	O	O
content	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
glycation-induced	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Also	NN	O	O
,	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analyses	NN	O	O
using	NN	O	O
specific	NN	O	O
antiserum	NN	O	O
revealed	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
an	NN	O	O
insulin	NN	O	B-protein
gene	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
PDX-1/IPF1/STF-1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
effects	NN	O	O
of	NN	O	O
D-ribose	NN	O	O
seemed	NN	O	O
almost	NN	O	O
irreversible	NN	O	O
but	NN	O	O
could	NN	O	O
be	NN	O	O
prevented	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
1	NN	O	O
mM	NN	O	O
aminoguanidine	NN	O	O
or	NN	O	O
10	NN	O	O
mM	NN	O	O
N-acetylcysteine	NN	O	O
,	NN	O	O
thus	NN	O	O
suggesting	NN	O	O
that	NN	O	O
glycation	NN	O	O
and	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
,	NN	O	O
generated	NN	O	O
through	NN	O	O
the	NN	O	O
glycation	NN	O	O
reaction	NN	O	O
,	NN	O	O
serve	NN	O	O
as	NN	O	O
mediators	NN	O	O
of	NN	O	O
the	NN	O	O
phenomena	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
protein	NN	O	O
glycation	NN	O	O
in	NN	O	O
pancreatic	NN	O	B-cell_type
beta	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
occurs	NN	O	O
in	NN	O	O
vivo	NN	O	O
under	NN	O	O
chronic	NN	O	O
hyperglycemia	NN	O	O
,	NN	O	O
suppresses	NN	O	O
insulin	NN	O	O
gene	NN	O	O
transcription	NN	O	O
and	NN	O	O
thus	NN	O	O
can	NN	O	O
explain	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
beta	NN	O	O
cell	NN	O	O
glucose	NN	O	O
toxicity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
rheumatic	NN	O	O
diseases	NN	O	O
:	NN	O	O
a	NN	O	O
preliminary	NN	O	O
report	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
The	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
regulates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
many	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
or	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
.	NN	O	O

The	NN	O	O
aim	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
distinctive	NN	O	O
patterns	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
are	NN	O	O
seen	NN	O	O
in	NN	O	O
different	NN	O	O
forms	NN	O	O
of	NN	O	O
joint	NN	O	O
disease	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
The	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
these	NN	O	O
nucleoproteins	NN	O	B-protein
was	NN	O	O
examined	NN	O	O
in	NN	O	O
purified	NN	O	O
synovial	NN	O	B-cell_type
and	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
various	NN	O	O
chronic	NN	O	O
rheumatic	NN	O	O
diseases	NN	O	O
(	NN	O	O
12	NN	O	O
:	NN	O	O
four	NN	O	O
with	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
;	NN	O	O
five	NN	O	O
with	NN	O	O
spondyloarthropathies	NN	O	O
;	NN	O	O
and	NN	O	O
three	NN	O	O
with	NN	O	O
osteoarthritis	NN	O	O
)	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
disclosed	NN	O	O
two	NN	O	O
specific	NN	O	O
complexes	NN	O	O
bound	NN	O	O
to	NN	O	O
a	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
specific	NN	O	O
32P-labelled	NN	O	O
oligonucleotide	NN	O	O
in	NN	O	O
nucleoproteins	NN	O	B-protein
extracted	NN	O	O
from	NN	O	O
purified	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
synovial	NN	O	O
fluid	NN	O	O
and	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
.	NN	O	O

The	NN	O	O
complexes	NN	O	B-protein
consisted	NN	O	O
of	NN	O	O
p50/p50	NN	O	B-protein
homodimers	NN	O	I-protein
and	NN	O	O
p50/p65	NN	O	O
heterodimers	NN	O	O
.	NN	O	O

Increased	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
DNA	NN	O	O
in	NN	O	O
synovial	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
observed	NN	O	O
relative	NN	O	O
to	NN	O	O
peripheral	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
non-rheumatoid	NN	O	O
arthritis	NN	O	O
,	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
synovial	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
exclusively	NN	O	O
mediated	NN	O	O
by	NN	O	O
p50/p50	NN	O	B-protein
homodimers	NN	O	I-protein
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Overall	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
central	NN	O	O
part	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
infiltrating	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
chronic	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
is	NN	O	O
qualitatively	NN	O	O
different	NN	O	O
in	NN	O	O
rheumatoid	NN	O	B-cell_type
synovial	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
that	NN	O	O
in	NN	O	O
other	NN	O	O
forms	NN	O	O
of	NN	O	O
non-rheumatoid	NN	O	O
arthritis	NN	O	O
(	NN	O	O
for	NN	O	O
example	NN	O	O
,	NN	O	O
osteoarthritis	NN	O	O
,	NN	O	O
spondyloarthropathies	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

X	NN	O	B-DNA
chromosome	NN	O	I-DNA
inactivation	NN	O	O
patterns	NN	O	O
in	NN	O	O
normal	NN	O	O
females	NN	O	O
.	NN	O	O

Since	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
X	NN	O	B-DNA
chromosomes	NN	O	I-DNA
is	NN	O	O
randomly	NN	O	O
inactivated	NN	O	O
at	NN	O	O
an	NN	O	O
early	NN	O	O
stage	NN	O	O
of	NN	O	O
female	NN	O	O
embryonic	NN	O	O
development	NN	O	O
,	NN	O	O
X-linked	NN	O	B-protein
markers	NN	O	I-protein
have	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
origin	NN	O	O
and	NN	O	O
development	NN	O	O
of	NN	O	O
various	NN	O	O
neoplastic	NN	O	O
disorders	NN	O	O
in	NN	O	O
affected	NN	O	O
heterozygous	NN	O	O
women	NN	O	O
;	NN	O	O
clonality	NN	O	O
assays	NN	O	O
have	NN	O	O
provided	NN	O	O
a	NN	O	O
useful	NN	O	O
tool	NN	O	O
to	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
neoplasia	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
a	NN	O	O
technique	NN	O	O
of	NN	O	O
clonal	NN	O	O
analysis	NN	O	O
has	NN	O	O
been	NN	O	O
devised	NN	O	O
that	NN	O	O
takes	NN	O	O
advantage	NN	O	O
of	NN	O	O
a	NN	O	O
highly	NN	O	O
polymorphic	NN	O	O
short	NN	O	B-DNA
tandem	NN	O	I-DNA
repeat	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
X-linked	NN	O	B-DNA
human	NN	O	I-DNA
androgen	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
AR	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
heterozygosity	NN	O	O
rate	NN	O	O
approaching	NN	O	O
90	NN	O	O
%	NN	O	O
.	NN	O	O

The	NN	O	O
rapid	NN	O	O
expansion	NN	O	O
of	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
women	NN	O	O
now	NN	O	O
suitable	NN	O	O
for	NN	O	O
X	NN	O	O
inactivation	NN	O	O
analysis	NN	O	O
has	NN	O	O
however	NN	O	O
given	NN	O	O
rise	NN	O	O
to	NN	O	O
new	NN	O	O
controversies	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
more	NN	O	O
troublesome	NN	O	O
being	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
modification	NN	O	O
of	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
X-	NN	O	B-DNA
chromosome	NN	O	I-DNA
inactivation	NN	O	O
pattern	NN	O	O
in	NN	O	O
blood	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
elderly	NN	O	O
women	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
analyze	NN	O	O
with	NN	O	O
the	NN	O	O
AR	NN	O	O
assay	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
166	NN	O	O
healthy	NN	O	O
females	NN	O	O
aged	NN	O	O
between	NN	O	O
8	NN	O	O
and	NN	O	O
94	NN	O	O
years	NN	O	O
,	NN	O	O
with	NN	O	O
no	NN	O	O
history	NN	O	O
of	NN	O	O
genetic	NN	O	O
or	NN	O	O
neoplastic	NN	O	O
familial	NN	O	O
disorders	NN	O	O
.	NN	O	O

We	NN	O	O
failed	NN	O	O
to	NN	O	O
find	NN	O	O
any	NN	O	O
correlation	NN	O	O
between	NN	O	O
age	NN	O	O
and	NN	O	O
X-	NN	O	B-DNA
chromosome	NN	O	I-DNA
inactivation	NN	O	O
pattern	NN	O	O
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.17	NN	O	O
)	NN	O	O
,	NN	O	O
even	NN	O	O
subdividing	NN	O	O
the	NN	O	O
subjects	NN	O	O
in	NN	O	O
different	NN	O	O
age	NN	O	O
groups	NN	O	O
according	NN	O	O
to	NN	O	O
the	NN	O	O
criteria	NN	O	O
used	NN	O	O
by	NN	O	O
other	NN	O	O
researchers	NN	O	O
,	NN	O	O
and	NN	O	O
therefore	NN	O	O
reaffirm	NN	O	O
that	NN	O	O
,	NN	O	O
when	NN	O	O
tested	NN	O	O
for	NN	O	O
with	NN	O	O
well-standardized	NN	O	O
and	NN	O	O
accurate	NN	O	O
criteria	NN	O	O
,	NN	O	O
extremely	NN	O	O
unbalanced	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
chromosome	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
truly	NN	O	O
uncommon	NN	O	O
phenomenon	NN	O	O
in	NN	O	O
normal	NN	O	O
women	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
Stat4	NN	O	B-protein
in	NN	O	O
species-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
Th	NN	O	O
cell	NN	O	O
development	NN	O	O
by	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
IFNs	NN	O	I-protein
.	NN	O	O

Type	NN	O	B-protein
I	NN	O	I-protein
IFNs	NN	O	I-protein
(	NN	O	O
IFN-alpha/beta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
mouse	NN	O	O
,	NN	O	O
Th	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
IFN-alpha/beta	NN	O	B-protein
act	NN	O	O
directly	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
drive	NN	O	O
Th1	NN	O	O
development	NN	O	O
,	NN	O	O
bypassing	NN	O	O
the	NN	O	O
need	NN	O	O
for	NN	O	O
IL-12	NN	O	B-protein
-induced	NN	O	O
signaling	NN	O	O
,	NN	O	O
whereas	NN	O	O
IFN-alpha	NN	O	B-protein
can	NN	O	O
not	NN	O	O
substitute	NN	O	O
IL-12	NN	O	B-protein
for	NN	O	O
mouse	NN	O	O
Th1	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
this	NN	O	O
species	NN	O	O
specificity	NN	O	O
is	NN	O	O
that	NN	O	O
IFN-alpha/beta	NN	O	B-protein
activate	NN	O	O
Stat4	NN	O	B-protein
in	NN	O	O
differentiating	NN	O	O
human	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
mouse	NN	O	O
,	NN	O	O
Th	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Unlike	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
which	NN	O	O
acts	NN	O	O
only	NN	O	O
on	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
IFN-alpha/beta	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
Stat4	NN	O	B-protein
not	NN	O	O
only	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
Th1	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
Th2	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
restimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
Th2	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
clones	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IFN-alpha	NN	O	B-protein
does	NN	O	O
not	NN	O	O
induce	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat4	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
polarized	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
induce	NN	O	O
phenotype	NN	O	O
reversal	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
Th2	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
cloning	NN	O	O
of	NN	O	O
FKHRL1P2	NN	O	B-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
developmentally	NN	O	O
regulated	NN	O	O
fork	NN	O	B-protein
head	NN	O	I-protein
domain	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
fork	NN	O	B-protein
head	NN	O	I-protein
domain	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
cloned	NN	O	O
and	NN	O	O
characterized	NN	O	O
a	NN	O	O
fork	NN	O	B-DNA
head	NN	O	I-DNA
cDNA	NN	O	I-DNA
from	NN	O	O
human	NN	O	B-RNA
T	NN	O	I-RNA
helper	NN	O	I-RNA
cell	NN	O	I-RNA
mRNA	NN	O	I-RNA
using	NN	O	O
differential	NN	O	O
display	NN	O	O
RT-PCR	NN	O	O
.	NN	O	O

The	NN	O	O
cDNA	NN	O	O
contains	NN	O	O
a	NN	O	O
546-nucleotide	NN	O	B-DNA
(	NN	O	I-DNA
nt	NN	O	I-DNA
)	NN	O	I-DNA
open	NN	O	I-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
(	NN	O	O
ORF	NN	O	B-DNA
)	NN	O	O
that	NN	O	O
codes	NN	O	O
for	NN	O	O
the	NN	O	O
carboxyl-terminal	NN	O	B-protein
180	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
(	NN	O	O
aa	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
recently	NN	O	O
identified	NN	O	O
fkhrl1	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
ORF	NN	O	B-DNA
does	NN	O	O
not	NN	O	O
contain	NN	O	O
the	NN	O	O
characteristic	NN	O	O
DNA-binding	NN	O	B-DNA
domain	NN	O	I-DNA
found	NN	O	O
in	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
forkhead	NN	O	B-protein
protein	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

In-vitro	NN	O	O
transcription/translation	NN	O	O
of	NN	O	O
this	NN	O	O
cDNA	NN	O	B-DNA
expressed	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
approximately	NN	O	O
20	NN	O	O
kDa	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
generated	NN	O	O
antibodies	NN	O	O
that	NN	O	O
specifically	NN	O	O
immunoprecipitated	NN	O	O
the	NN	O	O
in-vitro-translated	NN	O	B-protein
20-kDa	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

This	NN	O	O
antibody	NN	O	O
also	NN	O	O
recognizes	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
a	NN	O	O
70-kDa	NN	O	O
protein	NN	O	O
corresponding	NN	O	O
in	NN	O	O
size	NN	O	O
to	NN	O	O
that	NN	O	O
predicted	NN	O	O
for	NN	O	O
the	NN	O	O
fkhrl1	NN	O	B-DNA
gene	NN	O	I-DNA
product	NN	O	O
.	NN	O	O

The	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
for	NN	O	O
fkhrl1	NN	O	B-DNA
is	NN	O	O
elevated	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
helper-induced	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
comparison	NN	O	O
to	NN	O	O
PHA-stimulated	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Further	NN	O	O
characterization	NN	O	O
of	NN	O	O
FKHRL1	NN	O	B-DNA
and	NN	O	O
its	NN	O	O
related	NN	O	O
family	NN	O	O
members	NN	O	O
should	NN	O	O
shed	NN	O	O
light	NN	O	O
on	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
this	NN	O	O
fork	NN	O	B-DNA
head	NN	O	I-DNA
gene	NN	O	I-DNA
subfamily	NN	O	I-DNA
and	NN	O	O
their	NN	O	O
role	NN	O	O
in	NN	O	O
T	NN	O	O
helper	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
Th1	NN	O	O
development	NN	O	O
mediated	NN	O	O
by	NN	O	O
GATA-3	NN	O	B-protein
through	NN	O	O
an	NN	O	O
IL-4-independent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-3	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
selectively	NN	O	O
expressed	NN	O	O
in	NN	O	O
Th2	NN	O	O
but	NN	O	O
not	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
to	NN	O	O
augment	NN	O	O
Th2-specific	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
loss	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
developing	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
through	NN	O	O
Stat4	NN	O	B-protein
and	NN	O	O
does	NN	O	O
not	NN	O	O
simply	NN	O	O
result	NN	O	O
from	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
novel	NN	O	O
role	NN	O	O
for	NN	O	O
GATA-3	NN	O	B-protein
in	NN	O	O
directly	NN	O	O
repressing	NN	O	O
Th1	NN	O	O
development	NN	O	O
distinct	NN	O	O
from	NN	O	O
its	NN	O	O
positive	NN	O	O
actions	NN	O	O
on	NN	O	O
Th2-specific	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

GATA-3	NN	O	B-protein
inhibits	NN	O	O
Th1	NN	O	B-protein
cytokines	NN	O	I-protein
by	NN	O	O
a	NN	O	O
cell-intrinsic	NN	O	O
mechanism	NN	O	O
that	NN	O	O
is	NN	O	O
not	NN	O	O
dependent	NN	O	O
on	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
that	NN	O	O
may	NN	O	O
involve	NN	O	O
repression	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
GATA-3	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
IL-12	NN	O	B-protein
signaling	NN	O	O
are	NN	O	O
mutually	NN	O	O
antagonistic	NN	O	O
,	NN	O	O
which	NN	O	O
facilitates	NN	O	O
rapid	NN	O	O
dominance	NN	O	O
of	NN	O	O
one	NN	O	O
pathway	NN	O	O
during	NN	O	O
early	NN	O	O
Th	NN	O	O
development	NN	O	O
,	NN	O	O
producing	NN	O	O
a	NN	O	O
stable	NN	O	O
divergence	NN	O	O
in	NN	O	O
cytokine	NN	O	B-protein
profiles	NN	O	O
.	NN	O	O

-DOCSTART-	O

Conserved	NN	O	B-DNA
elements	NN	O	I-DNA
containing	NN	O	O
NF-E2	NN	O	B-DNA
and	NN	O	I-DNA
tandem	NN	O	I-DNA
GATA	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
erythroid-specific	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
reorganization	NN	O	O
within	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
locus	NN	O	I-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Proper	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
gene	NN	O	I-DNA
locus	NN	O	I-DNA
requires	NN	O	O
the	NN	O	O
associated	NN	O	O
locus	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
LCR	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Structurally	NN	O	O
,	NN	O	O
the	NN	O	O
LCR	NN	O	B-DNA
is	NN	O	O
defined	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
four	NN	O	O
domains	NN	O	O
of	NN	O	O
erythroid-specific	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
.	NN	O	O

These	NN	O	O
domains	NN	O	O
,	NN	O	O
which	NN	O	O
have	NN	O	O
been	NN	O	O
characterized	NN	O	O
as	NN	O	O
DNase	NN	O	B-DNA
I	NN	O	I-DNA
hypersensitive	NN	O	I-DNA
sites	NN	O	I-DNA
(	NN	O	O
HSs	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
comprise	NN	O	O
the	NN	O	O
active	NN	O	O
elements	NN	O	O
of	NN	O	O
the	NN	O	O
LCR	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
major	NN	O	O
focus	NN	O	O
of	NN	O	O
this	NN	O	O
research	NN	O	O
is	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
cis	NN	O	B-DNA
-acting	NN	O	I-DNA
elements	NN	O	I-DNA
which	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
these	NN	O	O
domains	NN	O	O
of	NN	O	O
unique	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
investigations	NN	O	O
on	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
LCR	NN	O	B-DNA
HS4	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
NF-E2	NN	O	B-DNA
and	NN	O	I-DNA
tandem	NN	O	I-DNA
,	NN	O	I-DNA
inverted	NN	O	I-DNA
GATA	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
native	NN	O	B-DNA
HS	NN	O	I-DNA
.	NN	O	O

Similarly	NN	O	O
arranged	NN	O	O
NF-E2	NN	O	B-DNA
and	NN	O	I-DNA
tandem	NN	O	I-DNA
GATA	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
present	NN	O	O
within	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
other	NN	O	O
human	NN	O	B-DNA
LCR	NN	O	I-DNA
HSs	NN	O	I-DNA
and	NN	O	O
are	NN	O	O
evolutionarily	NN	O	O
conserved	NN	O	O
.	NN	O	O

Using	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
HSs	NN	O	I-DNA
2	NN	O	I-DNA
and	NN	O	I-DNA
3	NN	O	I-DNA
we	NN	O	O
have	NN	O	O
tested	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
these	NN	O	O
NF-E2	NN	O	B-DNA
and	NN	O	I-DNA
GATA	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
common	NN	O	O
requirements	NN	O	O
for	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
all	NN	O	O
LCR	NN	O	B-DNA
HSs	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
mutation	NN	O	O
of	NN	O	O
these	NN	O	O
elements	NN	O	O
,	NN	O	O
and	NN	O	O
particularly	NN	O	O
the	NN	O	O
GATA	NN	O	B-DNA
elements	NN	O	I-DNA
,	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
decrease	NN	O	O
or	NN	O	O
complete	NN	O	O
loss	NN	O	O
of	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
hypersensitivity	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
imply	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
common	NN	O	O
structural	NN	O	O
elements	NN	O	O
within	NN	O	O
the	NN	O	O
core	NN	O	O
of	NN	O	O
each	NN	O	O
LCR	NN	O	B-DNA
HS	NN	O	I-DNA
which	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
erythroid-specific	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
reorganization	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
of	NN	O	O
ARE-containing	NN	O	B-DNA
genes	NN	O	I-DNA
by	NN	O	O
AU-motif-directed	NN	O	O
display	NN	O	O
.	NN	O	O

A	NN	O	O
procedure	NN	O	O
suitable	NN	O	O
for	NN	O	O
cloning	NN	O	B-RNA
labile	NN	O	I-RNA
mRNAs	NN	O	I-RNA
that	NN	O	O
contain	NN	O	O
AU	NN	O	O
motifs	NN	O	O
is	NN	O	O
presented	NN	O	O
(	NN	O	O
AU-DD	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
motifs	NN	O	O
are	NN	O	O
regulatory	NN	O	B-DNA
sequences	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
so-called	NN	O	O
AU-rich	NN	O	B-DNA
elements	NN	O	I-DNA
(	NN	O	O
AREs	NN	O	B-DNA
)	NN	O	O
often	NN	O	O
found	NN	O	O
in	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
untranslated	NN	O	I-DNA
regions	NN	O	I-DNA
of	NN	O	O
genes	NN	O	O
such	NN	O	O
as	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
proto-oncogenes	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

AU-DD	NN	O	B-DNA
is	NN	O	O
an	NN	O	O
AU-motif-directed	NN	O	B-DNA
differential	NN	O	I-DNA
display	NN	O	I-DNA
that	NN	O	O
permits	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
ARE-containing	NN	O	B-DNA
genes	NN	O	I-DNA
differentially	NN	O	O
expressed	NN	O	O
after	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
applied	NN	O	O
to	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
a	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
to	NN	O	O
isolate	NN	O	O
59	NN	O	B-DNA
cDNA	NN	O	I-DNA
fragments	NN	O	I-DNA
associated	NN	O	O
to	NN	O	O
activation	NN	O	O
.	NN	O	O

Fourteen	NN	O	O
percent	NN	O	O
of	NN	O	O
isolated	NN	O	O
fragments	NN	O	O
belong	NN	O	O
to	NN	O	O
already	NN	O	O
known	NN	O	O
genes	NN	O	O
that	NN	O	O
certainly	NN	O	O
are	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
transduction/transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
remaining	NN	O	O
86	NN	O	O
%	NN	O	O
correspond	NN	O	O
to	NN	O	O
unknown	NN	O	O
genes	NN	O	B-DNA
of	NN	O	O
which	NN	O	O
92	NN	O	O
%	NN	O	O
have	NN	O	O
been	NN	O	O
confirmed	NN	O	O
to	NN	O	O
be	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
efficiency	NN	O	O
of	NN	O	O
the	NN	O	O
system	NN	O	O
and	NN	O	O
support	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
numerous	NN	O	O
genes	NN	O	B-DNA
falling	NN	O	O
into	NN	O	O
those	NN	O	O
categories	NN	O	O
remain	NN	O	O
unidentified	NN	O	O
and	NN	O	O
that	NN	O	O
they	NN	O	O
can	NN	O	O
be	NN	O	O
cloned	NN	O	O
by	NN	O	O
this	NN	O	O
method	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	O
factor	NN	O	O
NF-kappaB	NN	O	B-protein
regulation	NN	O	O
of	NN	O	O
renal	NN	O	O
fibrosis	NN	O	O
during	NN	O	O
ureteral	NN	O	O
obstruction	NN	O	O
.	NN	O	O

Irrespective	NN	O	O
of	NN	O	O
the	NN	O	O
etiology	NN	O	O
,	NN	O	O
many	NN	O	O
kidney	NN	O	O
diseases	NN	O	O
result	NN	O	O
in	NN	O	O
inflammation	NN	O	O
and	NN	O	O
fibrosis	NN	O	O
of	NN	O	O
the	NN	O	O
tubulointerstitium	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
subsequent	NN	O	O
loss	NN	O	O
of	NN	O	O
renal	NN	O	O
function	NN	O	O
.	NN	O	O

To	NN	O	O
initiate	NN	O	O
any	NN	O	O
disease	NN	O	O
process	NN	O	O
or	NN	O	O
for	NN	O	O
any	NN	O	O
disease	NN	O	O
process	NN	O	O
to	NN	O	O
progress	NN	O	O
,	NN	O	O
there	NN	O	O
must	NN	O	O
be	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
within	NN	O	O
the	NN	O	O
affected	NN	O	O
tissue	NN	O	O
.	NN	O	O

The	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	I-protein
NF-kappaB	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
regulates	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
inflammation	NN	O	O
,	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
and	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

This	NN	O	O
review	NN	O	O
discusses	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
in	NN	O	O
general	NN	O	O
and	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
with	NN	O	O
cellular/molecular	NN	O	O
events	NN	O	O
of	NN	O	O
renal	NN	O	O
inflammation	NN	O	O
and	NN	O	O
fibrosis	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
during	NN	O	O
sodium	NN	O	O
phenylacetate	NN	O	O
induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

During	NN	O	O
15	NN	O	O
days	NN	O	O
of	NN	O	O
treatment	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
sodium	NN	O	O
phenylacetate	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
hemoglobin	NN	O	B-protein
concentration	NN	O	O
with	NN	O	O
a	NN	O	O
similar	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
gamma-globin	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Morphological	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
characteristic	NN	O	O
features	NN	O	O
of	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
and	NN	O	O
maturation	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
same	NN	O	O
time	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
antigenes	NN	O	I-protein
CD33	NN	O	B-protein
,	NN	O	O
CD34	NN	O	B-protein
,	NN	O	O
CD45	NN	O	B-protein
,	NN	O	O
CD71	NN	O	B-protein
and	NN	O	O
glycophorin	NN	O	B-protein
A	NN	O	I-protein
.	NN	O	O

Likewise	NN	O	O
,	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
erythroid	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
GATA-1	NN	O	B-protein
,	NN	O	O
GATA-2	NN	O	B-protein
,	NN	O	O
NF-E2	NN	O	B-protein
,	NN	O	O
SCL	NN	O	B-protein
and	NN	O	O
RBTN2	NN	O	B-protein
,	NN	O	O
all	NN	O	O
expressed	NN	O	O
in	NN	O	O
untreated	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
did	NN	O	O
not	NN	O	O
increase	NN	O	O
during	NN	O	O
sodium	NN	O	O
phenylacetate	NN	O	O
induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
Evi-1	NN	O	B-protein
and	NN	O	O
c-myb	NN	O	B-protein
,	NN	O	O
known	NN	O	O
to	NN	O	O
inhibit	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
,	NN	O	O
did	NN	O	O
not	NN	O	O
decrease	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
sodium	NN	O	O
phenylacetate	NN	O	O
treatment	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
increases	NN	O	O
gamma-globin	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
induces	NN	O	O
cell	NN	O	O
maturation	NN	O	O
as	NN	O	O
judged	NN	O	O
by	NN	O	O
morphology	NN	O	O
without	NN	O	O
affecting	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
erythroid	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
some	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
beta-like	NN	O	B-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Isolation	NN	O	O
of	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	B-DNA
promoter	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
human	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
.	NN	O	O

The	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	I-protein
MHC	NN	O	I-protein
)	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
.	NN	O	O

The	NN	O	O
tissular	NN	O	O
patterns	NN	O	O
of	NN	O	O
CIITA	NN	O	O
and	NN	O	O
MHC	NN	O	O
class	NN	O	O
II	NN	O	O
gene	NN	O	O
expression	NN	O	O
are	NN	O	O
tightly	NN	O	O
correlated	NN	O	O
:	NN	O	O
CIITA	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
highly	NN	O	O
expressed	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
in	NN	O	O
macrophage	NN	O	B-cell_line
and	NN	O	I-cell_line
epithelial	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
first	NN	O	O
isolated	NN	O	O
two	NN	O	O
overlapping	NN	O	O
cosmids	NN	O	B-DNA
encoding	NN	O	O
human	NN	O	B-protein
CIITA	NN	O	I-protein
which	NN	O	O
,	NN	O	O
when	NN	O	O
co-transfected	NN	O	O
,	NN	O	O
are	NN	O	O
able	NN	O	O
to	NN	O	O
restore	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
a	NN	O	O
B-lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
B-LCL	NN	O	B-cell_line
)	NN	O	O
defective	NN	O	O
for	NN	O	O
CIITA	NN	O	B-protein
.	NN	O	O

Subsequently	NN	O	O
,	NN	O	O
a	NN	O	O
1.8	NN	O	B-DNA
kilobase	NN	O	I-DNA
(	NN	O	I-DNA
kb	NN	O	I-DNA
)	NN	O	I-DNA
fragment	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CIITA	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
isolated	NN	O	O
and	NN	O	O
sequenced	NN	O	O
.	NN	O	O

A	NN	O	O
motif	NN	O	O
presenting	NN	O	O
a	NN	O	O
strong	NN	O	O
similarity	NN	O	O
to	NN	O	O
an	NN	O	O
initiator	NN	O	O
was	NN	O	O
detected	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
putative	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
GATA-2	NN	O	B-protein
,	NN	O	O
LyF-1	NN	O	B-protein
,	NN	O	O
ets-1	NN	O	B-protein
,	NN	O	O
AP1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
MZF1	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
two	NN	O	O
GAS	NN	O	B-DNA
motifs	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
introduced	NN	O	O
in	NN	O	O
front	NN	O	O
of	NN	O	O
a	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
this	NN	O	O
promoter	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
direct	NN	O	O
a	NN	O	O
high	NN	O	O
luciferase	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B-LCL	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
luciferase	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
not	NN	O	O
stimulated	NN	O	O
after	NN	O	O
IFN-gamma	NN	O	B-protein
treatment	NN	O	O
when	NN	O	O
the	NN	O	O
construct	NN	O	O
was	NN	O	O
transfected	NN	O	O
in	NN	O	O
macrophage	NN	O	O
or	NN	O	O
in	NN	O	O
epithelial	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
an	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CIITA	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
observed	NN	O	O
in	NN	O	O
mouse	NN	O	B-cell_line
macrophage	NN	O	I-cell_line
and	NN	O	I-cell_line
fibrosarcoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
when	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
cosmid	NN	O	B-DNA
containing	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CIITA	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
lacking	NN	O	O
the	NN	O	O
1.8	NN	O	B-DNA
kb	NN	O	I-DNA
promoter	NN	O	I-DNA
described	NN	O	O
above	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
an	NN	O	O
intragenic	NN	O	O
promoter	NN	O	O
driving	NN	O	O
an	NN	O	O
IFN-gamma	NN	O	B-protein
-inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Phenotypic	NN	O	O
and	NN	O	O
functional	NN	O	O
studies	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
in	NN	O	O
human	NN	O	O
endometrium	NN	O	O
and	NN	O	O
endometriosis	NN	O	O
.	NN	O	O

The	NN	O	O
aetiology	NN	O	O
of	NN	O	O
endometriosis	NN	O	O
,	NN	O	O
a	NN	O	O
common	NN	O	O
and	NN	O	O
disabling	NN	O	O
disorder	NN	O	O
,	NN	O	O
is	NN	O	O
presently	NN	O	O
unknown	NN	O	O
,	NN	O	O
although	NN	O	O
immune	NN	O	O
dysfunction	NN	O	O
could	NN	O	O
allow	NN	O	O
ectopic	NN	O	O
endometrial	NN	O	O
fragments	NN	O	O
to	NN	O	O
survive	NN	O	O
outside	NN	O	O
the	NN	O	O
uterine	NN	O	O
cavity	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
investigate	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
leukocyte	NN	O	B-cell_type
populations	NN	O	I-cell_type
,	NN	O	O
steroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
,	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
eutopic	NN	O	O
and	NN	O	O
ectopic	NN	O	O
endometrium	NN	O	O
from	NN	O	O
women	NN	O	O
with	NN	O	O
endometriosis	NN	O	O
or	NN	O	O
adenomyosis	NN	O	O
at	NN	O	O
different	NN	O	O
phases	NN	O	O
of	NN	O	O
the	NN	O	O
menstrual	NN	O	O
cycle	NN	O	O
.	NN	O	O

Significantly	NN	O	O
increased	NN	O	O
oestrogen	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
numbers	NN	O	O
of	NN	O	O
CD8+	NN	O	O
leukocytes	NN	O	B-cell_type
were	NN	O	O
found	NN	O	O
in	NN	O	O
ectopic	NN	O	O
compared	NN	O	O
with	NN	O	O
eutopic	NN	O	O
endometrium	NN	O	O
in	NN	O	O
endometriosis	NN	O	O
,	NN	O	O
and	NN	O	O
CD56+	NN	O	O
endometrial	NN	O	O
granulated	NN	O	O
lymphocytes	NN	O	O
(	NN	O	O
eGLs	NN	O	O
)	NN	O	O
were	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
in	NN	O	O
ectopic	NN	O	O
endometrium	NN	O	O
.	NN	O	O

Apoptotic	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
rarely	NN	O	O
found	NN	O	O
in	NN	O	O
control	NN	O	O
and	NN	O	O
subject	NN	O	O
endometria	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
with	NN	O	O
endometriosis	NN	O	O
,	NN	O	O
adenomyotic	NN	O	O
lesions	NN	O	O
showed	NN	O	O
identical	NN	O	O
steroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
,	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
leukocyte	NN	O	O
subpopulations	NN	O	O
to	NN	O	O
eutopic	NN	O	O
endometrium	NN	O	O
,	NN	O	O
indicating	NN	O	O
different	NN	O	O
aetiologies	NN	O	O
for	NN	O	O
these	NN	O	O
disorders	NN	O	O
.	NN	O	O

The	NN	O	O
unusual	NN	O	O
CD56+	NN	O	B-cell_type
CD16-	NN	O	I-cell_type
eGLs	NN	O	I-cell_type
present	NN	O	O
in	NN	O	O
large	NN	O	O
numbers	NN	O	O
in	NN	O	O
late	NN	O	O
secretory	NN	O	O
phase	NN	O	O
eutopic	NN	O	O
endometrium	NN	O	O
were	NN	O	O
highly	NN	O	O
purified	NN	O	O
(	NN	O	O
>	NN	O	O
98	NN	O	O
%	NN	O	O
)	NN	O	O
by	NN	O	O
immunomagnetic	NN	O	O
separation	NN	O	O
.	NN	O	O

Except	NN	O	O
for	NN	O	O
a	NN	O	O
negligible	NN	O	O
cytotoxic	NN	O	O
activity	NN	O	O
of	NN	O	O
eGLs	NN	O	B-cell_type
from	NN	O	O
early	NN	O	O
proliferative	NN	O	O
samples	NN	O	O
,	NN	O	O
cytotoxic	NN	O	O
activity	NN	O	O
of	NN	O	O
eGLs	NN	O	B-cell_type
from	NN	O	O
non-pregnant	NN	O	O
endometrium	NN	O	O
during	NN	O	O
the	NN	O	O
menstrual	NN	O	O
cycle	NN	O	O
was	NN	O	O
comparable	NN	O	O
with	NN	O	O
those	NN	O	O
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
,	NN	O	O
predominantly	NN	O	O
CD56+	NN	O	B-cell_type
CD16+	NN	O	I-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

eGLs	NN	O	B-cell_type
from	NN	O	O
non-pregnant	NN	O	O
endometrium	NN	O	O
and	NN	O	O
early	NN	O	O
pregnancy	NN	O	O
showed	NN	O	O
a	NN	O	O
variable	NN	O	O
proliferative	NN	O	O
response	NN	O	O
to	NN	O	O
5	NN	O	O
and	NN	O	O
100	NN	O	O
U/ml	NN	O	O
interleukin-2	NN	O	B-protein
over	NN	O	O
48-h	NN	O	O
and	NN	O	O
120-h	NN	O	O
time	NN	O	O
courses	NN	O	O
.	NN	O	O

eGLs	NN	O	B-cell_type
are	NN	O	O
evidently	NN	O	O
functionally	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
eutopic	NN	O	O
endometrium	NN	O	O
.	NN	O	O

Their	NN	O	O
absence	NN	O	O
in	NN	O	O
endometriotic	NN	O	O
lesions	NN	O	O
together	NN	O	O
with	NN	O	O
increased	NN	O	O
CD+8	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
numbers	NN	O	O
and	NN	O	O
increased	NN	O	O
oestrogen	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
bcl-2	NN	O	B-protein
expression	NN	O	O
may	NN	O	O
have	NN	O	O
significant	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
development	NN	O	O
and	NN	O	O
progression	NN	O	O
of	NN	O	O
endometriosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
interleukin-1beta	NN	O	B-protein
transcription	NN	O	O
by	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
involves	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
latent	NN	O	B-protein
proteins	NN	O	I-protein
via	NN	O	O
their	NN	O	O
interaction	NN	O	O
with	NN	O	O
RBP	NN	O	B-protein
.	NN	O	O

Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
infects	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
outgrowth	NN	O	O
of	NN	O	O
immortalised	NN	O	B-cell_line
lymphoblastoid	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
LCLs	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
through	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
intracellular	NN	O	O
staining	NN	O	O
that	NN	O	O
interleukin-1beta	NN	O	B-protein
(	NN	O	O
IL-1beta	NN	O	B-protein
)	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
LCLs	NN	O	B-cell_line
and	NN	O	O
investigate	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
the	NN	O	O
individual	NN	O	O
latent	NN	O	B-protein
proteins	NN	O	I-protein
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
.	NN	O	O

Using	NN	O	O
RT-PCR	NN	O	O
,	NN	O	O
IL-1beta	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
up-regulated	NN	O	O
in	NN	O	O
EBV-transformed	NN	O	B-cell_line
LCLs	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
group	NN	O	B-cell_line
III	NN	O	I-cell_line
Burkitt	NN	O	I-cell_line
's	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
(	NN	O	I-cell_line
BL	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
compared	NN	O	O
with	NN	O	O
group	NN	O	B-cell_line
I	NN	O	I-cell_line
BL	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
IL-1beta	NN	O	B-protein
message	NN	O	O
could	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein-1	NN	O	I-protein
,	NN	O	O
EBV	NN	O	B-DNA
nuclear	NN	O	I-DNA
proteins	NN	O	I-DNA
2	NN	O	I-DNA
,	NN	O	I-DNA
3	NN	O	I-DNA
,	NN	O	I-DNA
4	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
6	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSAs	NN	O	O
)	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
-300	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1beta	NN	O	B-protein
promoter	NN	O	O
,	NN	O	O
which	NN	O	O
contains	NN	O	O
a	NN	O	O
nuclear	NN	O	B-DNA
factor-kappaB	NN	O	I-DNA
(	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
contained	NN	O	O
a	NN	O	O
functional	NN	O	B-DNA
RBP	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
RBP	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
could	NN	O	O
be	NN	O	O
inhibited	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
EBV	NN	O	B-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
3	NN	O	I-protein
and	NN	O	I-protein
6	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
proteins	NN	O	O
displace	NN	O	O
RBP	NN	O	B-protein
from	NN	O	O
its	NN	O	O
recognition	NN	O	B-DNA
sequence	NN	O	I-DNA
,	NN	O	O
removing	NN	O	O
transcriptional	NN	O	O
repression	NN	O	O
and	NN	O	O
allowing	NN	O	O
gene	NN	O	O
transcription	NN	O	O
to	NN	O	O
occur	NN	O	O
.	NN	O	O

In	NN	O	O
group	NN	O	B-cell_line
I	NN	O	I-cell_line
BL	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
containing	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
only	NN	O	O
RBP	NN	O	B-protein
binding	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
EMSAs	NN	O	O
,	NN	O	O
whereas	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
could	NN	O	O
be	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
EBV-transformed	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
containing	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
activated	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein-1	NN	O	I-protein
led	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
that	NN	O	O
was	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
the	NN	O	O
IL-1beta	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
EBV	NN	O	O
can	NN	O	O
up-regulate	NN	O	O
IL-1beta	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
possibly	NN	O	O
by	NN	O	O
using	NN	O	O
RBP	NN	O	B-protein
,	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
or	NN	O	O
both	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD27	NN	O	B-protein
/CD70	NN	O	O
interaction	NN	O	O
augments	NN	O	O
IgE	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
promoting	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
memory	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
switching	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
several	NN	O	O
signals	NN	O	O
given	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
cell	NN	O	O
contact-delivered	NN	O	O
signals	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
CD27	NN	O	B-protein
/CD70	NN	O	O
interaction	NN	O	O
in	NN	O	O
B	NN	O	O
cell	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
CD27	NN	O	O
ligand	NN	O	O
(	NN	O	O
CD70	NN	O	O
)	NN	O	O
transfectants	NN	O	O
to	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
increased	NN	O	O
the	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
synergistically	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
plus	NN	O	O
anti-CD40	NN	O	B-protein
mAb	NN	O	I-protein
(	NN	O	O
anti-CD40	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
CD70	NN	O	B-cell_line
transfectants	NN	O	I-cell_line
was	NN	O	O
dose	NN	O	O
dependent	NN	O	O
and	NN	O	O
was	NN	O	O
completely	NN	O	O
blocked	NN	O	O
by	NN	O	O
anti-CD70	NN	O	B-protein
mAb	NN	O	I-protein
.	NN	O	I-protein

CD27+	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
had	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
produce	NN	O	O
IgE	NN	O	B-protein
,	NN	O	O
which	NN	O	O
was	NN	O	O
increased	NN	O	O
by	NN	O	O
contact	NN	O	O
with	NN	O	O
CD70	NN	O	O
transfectants	NN	O	O
,	NN	O	O
whereas	NN	O	O
CD27-	NN	O	O
B	NN	O	O
cells	NN	O	O
did	NN	O	O
not	NN	O	O
produce	NN	O	O
IgE	NN	O	B-protein
.	NN	O	O

CD27	NN	O	B-protein
/CD70	NN	O	O
interaction	NN	O	O
enhanced	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
or	NN	O	O
IL-4	NN	O	B-protein
plus	NN	O	O
anti-CD40	NN	O	B-protein
.	NN	O	O

The	NN	O	O
augmentation	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
by	NN	O	O
CD70	NN	O	O
transfectants	NN	O	O
was	NN	O	O
apparent	NN	O	O
in	NN	O	O
CD27+	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
was	NN	O	O
mild	NN	O	O
in	NN	O	O
CD27-	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
helper	NN	O	O
activity	NN	O	O
for	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
by	NN	O	O
the	NN	O	O
CD27	NN	O	B-protein
/CD70	NN	O	O
interaction	NN	O	O
did	NN	O	O
not	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
enhancement	NN	O	O
of	NN	O	O
germline	NN	O	B-RNA
epsilon	NN	O	I-RNA
transcripts	NN	O	I-RNA
.	NN	O	O

Flow	NN	O	O
cytometric	NN	O	O
and	NN	O	O
morphological	NN	O	O
analyses	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
CD70	NN	O	O
transfectants	NN	O	O
to	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
remarkably	NN	O	O
promoted	NN	O	O
differentiation	NN	O	O
into	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
CD27	NN	O	B-protein
cross-linking	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
positive	NN	O	B-protein
regulatory	NN	O	I-protein
domain	NN	O	I-protein
I-binding	NN	O	I-protein
factor-1	NN	O	I-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
signaling	NN	O	O
via	NN	O	O
CD27	NN	O	B-protein
on	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
induces	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
,	NN	O	O
in	NN	O	O
cooperation	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
,	NN	O	O
by	NN	O	O
promoting	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
positive	NN	O	B-protein
regulatory	NN	O	I-protein
domain	NN	O	I-protein
I-binding	NN	O	I-protein
factor-1	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type-1	NN	O	O
transcription	NN	O	O
:	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
5'-untranslated	NN	O	O
leader	NN	O	O
region	NN	O	O
(	NN	O	O
review	NN	O	O
)	NN	O	O
.	NN	O	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type-1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
transcription	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
host-cell	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
with	NN	O	O
cis-regulatory	NN	O	B-DNA
DNA	NN	O	I-DNA
elements	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Much	NN	O	O
attention	NN	O	O
has	NN	O	O
focused	NN	O	O
on	NN	O	O
the	NN	O	O
series	NN	O	O
of	NN	O	O
sequence	NN	O	O
elements	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
U3	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
including	NN	O	O
the	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
transcribed	NN	O	O
5'-untranslated	NN	O	B-DNA
leader	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
5'-UTR	NN	O	B-DNA
)	NN	O	O
also	NN	O	O
contains	NN	O	O
important	NN	O	O
transcriptional	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
situated	NN	O	O
downstream	NN	O	O
of	NN	O	O
transcription	NN	O	O
interact	NN	O	O
with	NN	O	O
constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
mediate	NN	O	O
transmission	NN	O	O
of	NN	O	O
cellular	NN	O	O
activation	NN	O	O
signals	NN	O	O
,	NN	O	O
and	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
efficient	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
and	NN	O	O
replication	NN	O	O
.	NN	O	O

The	NN	O	O
5'-UTR	NN	O	B-DNA
contains	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP-1	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
IRF	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Sp1	NN	O	B-protein
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
these	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
can	NN	O	O
interfere	NN	O	O
with	NN	O	O
the	NN	O	O
viral	NN	O	O
response	NN	O	O
to	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
,	NN	O	O
decrease	NN	O	O
LTR	NN	O	B-DNA
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
inhibit	NN	O	O
viral	NN	O	O
replication	NN	O	O
.	NN	O	O

The	NN	O	O
5'-UTR	NN	O	B-DNA
also	NN	O	O
interacts	NN	O	O
with	NN	O	O
a	NN	O	O
specific	NN	O	O
nucleosome	NN	O	B-protein
that	NN	O	O
is	NN	O	O
rapidly	NN	O	O
displaced	NN	O	O
during	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
latent	NN	O	O
provirus	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
inducible	NN	O	B-DNA
transcription	NN	O	I-DNA
factor	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
5'-UTR	NN	O	B-DNA
comprise	NN	O	O
a	NN	O	O
downstream	NN	O	B-DNA
enhancer	NN	O	I-DNA
domain	NN	O	I-DNA
that	NN	O	O
can	NN	O	O
function	NN	O	O
independent	NN	O	O
of	NN	O	O
,	NN	O	O
or	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
,	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
promoter	NN	O	I-DNA
to	NN	O	O
rapidly	NN	O	O
increase	NN	O	O
latent	NN	O	O
proviral	NN	O	O
transcription	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
host-cell	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
that	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
5'-UTR	NN	O	B-DNA
and	NN	O	O
discuss	NN	O	O
their	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
PAK	NN	O	O
activation	NN	O	O
and	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
cytoskeleton	NN	O	O
by	NN	O	O
the	NN	O	O
linker	NN	O	B-protein
protein	NN	O	I-protein
SLP-76	NN	O	B-protein
.	NN	O	O

Tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
linker	NN	O	B-protein
proteins	NN	O	I-protein
enables	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
TCR	NN	O	I-protein
)	NN	O	I-protein
-associated	NN	O	I-protein
protein	NN	O	I-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
to	NN	O	O
phosphorylate	NN	O	O
and	NN	O	O
regulate	NN	O	O
effector	NN	O	B-protein
molecules	NN	O	I-protein
that	NN	O	O
generate	NN	O	O
second	NN	O	B-protein
messengers	NN	O	I-protein
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
SLP-76	NN	O	B-protein
linker	NN	O	O
protein	NN	O	O
interacts	NN	O	O
with	NN	O	O
both	NN	O	O
nck	NN	O	B-protein
,	NN	O	O
an	NN	O	O
adaptor	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
Vav	NN	O	B-protein
,	NN	O	O
a	NN	O	O
guanine	NN	O	B-protein
nucleotide	NN	O	I-protein
exchange	NN	O	I-protein
factor	NN	O	I-protein
for	NN	O	O
Rho-family	NN	O	B-protein
GTPases	NN	O	I-protein
.	NN	O	O

The	NN	O	O
assembly	NN	O	O
of	NN	O	O
this	NN	O	O
tri-molecular	NN	O	B-protein
complex	NN	O	I-protein
permits	NN	O	O
the	NN	O	O
activated	NN	O	O
Rho-family	NN	O	B-protein
GTPases	NN	O	I-protein
to	NN	O	O
regulate	NN	O	O
target	NN	O	O
effectors	NN	O	O
that	NN	O	O
interact	NN	O	O
through	NN	O	O
nck	NN	O	B-protein
.	NN	O	O

In	NN	O	O
turn	NN	O	O
,	NN	O	O
assembly	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
mediates	NN	O	O
the	NN	O	O
enzymatic	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
p21-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
facilitates	NN	O	O
actin	NN	O	B-protein
polymerization	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
linker	NN	O	B-protein
proteins	NN	O	I-protein
not	NN	O	O
only	NN	O	O
bridges	NN	O	O
the	NN	O	O
TCR-associated	NN	O	B-protein
PTK	NN	O	I-protein
,	NN	O	O
ZAP-70	NN	O	B-protein
,	NN	O	O
with	NN	O	O
downstream	NN	O	O
effector	NN	O	O
proteins	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
provides	NN	O	O
a	NN	O	O
scaffold	NN	O	O
to	NN	O	O
integrate	NN	O	O
distinct	NN	O	O
signaling	NN	O	B-protein
complexes	NN	O	I-protein
to	NN	O	O
regulate	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin-12	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
transformation	NN	O	O
with	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
.	NN	O	O

Although	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-12	NN	O	I-protein
was	NN	O	O
originally	NN	O	O
purified	NN	O	O
from	NN	O	O
an	NN	O	O
Epstein-Barr	NN	O	B-cell_line
(	NN	O	I-cell_line
EBV	NN	O	I-cell_line
)	NN	O	I-cell_line
-transformed	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
high	NN	O	O
correlation	NN	O	O
of	NN	O	O
EBV	NN	O	O
infection	NN	O	O
and	NN	O	O
IL-12	NN	O	B-protein
expression	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
,	NN	O	O
no	NN	O	O
study	NN	O	O
has	NN	O	O
reported	NN	O	O
whether	NN	O	O
EBV	NN	O	O
infection	NN	O	O
is	NN	O	O
directly	NN	O	O
linked	NN	O	O
to	NN	O	O
IL-12	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

To	NN	O	O
address	NN	O	O
this	NN	O	O
issue	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
IL-12	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
in	NN	O	O
vitro	NN	O	O
transformation	NN	O	O
with	NN	O	O
EBV	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
became	NN	O	O
capable	NN	O	O
of	NN	O	O
constitutively	NN	O	O
producing	NN	O	O
p40	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
transformation	NN	O	O
with	NN	O	O
EBV	NN	O	O
,	NN	O	O
coincident	NN	O	O
with	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
LMP1	NN	O	B-protein
)	NN	O	O
of	NN	O	O
EBV	NN	O	O
.	NN	O	O

These	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
expressed	NN	O	O
p40	NN	O	B-RNA
and	NN	O	I-RNA
p35	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
stimulation	NN	O	O
strongly	NN	O	O
enhanced	NN	O	O
p40	NN	O	O
and	NN	O	O
p70	NN	O	O
production	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
transfection	NN	O	O
with	NN	O	O
LMP1	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
into	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
Daudi	NN	O	B-cell_line
,	NN	O	O
led	NN	O	O
to	NN	O	O
p40	NN	O	B-protein
production	NN	O	O
with	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappaB	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
transformation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
EBV	NN	O	O
induces	NN	O	O
IL-12	NN	O	B-protein
expression	NN	O	O
potentially	NN	O	O
through	NN	O	O
LMP1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
human	NN	O	B-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
the	NN	O	O
lectin-type	NN	O	B-protein
oxidized	NN	O	I-protein
LDL	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
OLR1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
novel	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
natural	NN	O	B-protein
killer	NN	O	I-protein
gene	NN	O	I-protein
complex	NN	O	I-protein
with	NN	O	O
a	NN	O	O
unique	NN	O	O
expression	NN	O	O
profile	NN	O	O
.	NN	O	O

LOX-1	NN	O	B-protein
is	NN	O	O
an	NN	O	O
endothelial	NN	O	B-protein
receptor	NN	O	I-protein
for	NN	O	O
oxidized	NN	O	B-protein
low-density	NN	O	I-protein
lipoprotein	NN	O	I-protein
that	NN	O	O
plays	NN	O	O
essential	NN	O	O
roles	NN	O	O
in	NN	O	O
atherogenesis	NN	O	O
.	NN	O	O

LOX-1	NN	O	B-protein
has	NN	O	O
the	NN	O	O
highest	NN	O	O
homology	NN	O	O
with	NN	O	O
C-type	NN	O	B-protein
lectin	NN	O	I-protein
receptors	NN	O	I-protein
expressed	NN	O	O
on	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
cloned	NN	O	O
and	NN	O	O
characterized	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
LOX-1	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
HGMW-approved	NN	O	B-protein
symbol	NN	O	I-protein
OLR1	NN	O	I-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
gene	NN	O	O
structure	NN	O	O
of	NN	O	O
LOX-1	NN	O	B-protein
resembles	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
natural	NN	O	B-protein
killer	NN	O	I-protein
cell	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
and	NN	O	O
analyses	NN	O	O
of	NN	O	O
a	NN	O	O
yeast	NN	O	O
artificial	NN	O	O
chromosome	NN	O	O
contig	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
LOX-1	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
natural	NN	O	B-protein
killer	NN	O	I-protein
gene	NN	O	I-protein
complex	NN	O	I-protein
on	NN	O	O
chromosome	NN	O	B-DNA
12p12-p13	NN	O	I-DNA
,	NN	O	O
where	NN	O	O
the	NN	O	O
genes	NN	O	O
of	NN	O	O
the	NN	O	O
natural	NN	O	B-protein
killer	NN	O	I-protein
cell	NN	O	I-protein
receptors	NN	O	I-protein
cluster	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
LOX-1	NN	O	B-protein
is	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
natural	NN	O	B-protein
killer	NN	O	I-protein
cell	NN	O	I-protein
receptors	NN	O	I-protein
;	NN	O	O
LOX-1	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
vascular-rich	NN	O	O
organs	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

A	NN	O	O
1753-bp	NN	O	B-DNA
fragment	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
LOX-1	NN	O	B-DNA
gene	NN	O	I-DNA
had	NN	O	O
a	NN	O	O
functional	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

This	NN	O	O
region	NN	O	O
contains	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
the	NN	O	O
STAT	NN	O	B-protein
family	NN	O	I-protein
and	NN	O	O
NF-IL6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
LOX-1	NN	O	B-protein
was	NN	O	O
upregulated	NN	O	O
by	NN	O	O
several	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
LOX-1	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
new	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
natural	NN	O	B-protein
killer	NN	O	I-protein
gene	NN	O	I-protein
complex	NN	O	I-protein
with	NN	O	O
a	NN	O	O
unique	NN	O	O
expression	NN	O	O
profile	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
beta-casein	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
cytokines	NN	O	B-protein
:	NN	O	O
cross-talk	NN	O	O
between	NN	O	O
STAT5	NN	O	B-protein
and	NN	O	O
other	NN	O	O
signaling	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

The	NN	O	O
beta-casein	NN	O	B-DNA
promoter	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
widely	NN	O	O
used	NN	O	O
to	NN	O	O
monitor	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
(	NN	O	I-protein
signal	NN	O	I-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
)	NN	O	I-protein
5	NN	O	I-protein
since	NN	O	O
STAT5	NN	O	B-protein
was	NN	O	O
originally	NN	O	O
found	NN	O	O
as	NN	O	O
a	NN	O	O
mediator	NN	O	O
of	NN	O	O
PRL-inducible	NN	O	O
beta-casein	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
not	NN	O	O
only	NN	O	O
is	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
beta-casein	NN	O	B-DNA
gene	NN	O	I-DNA
regulated	NN	O	O
by	NN	O	O
STAT5	NN	O	B-protein
but	NN	O	O
it	NN	O	O
is	NN	O	O
also	NN	O	O
affected	NN	O	O
by	NN	O	O
other	NN	O	O
molecules	NN	O	O
such	NN	O	O
as	NN	O	O
glucocorticoid	NN	O	O
and	NN	O	O
Ras	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
beta-casein	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
beta-casein	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
cytotoxic	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
CTLL-2	NN	O	B-cell_line
,	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
activates	NN	O	O
STAT5	NN	O	B-protein
.	NN	O	O

While	NN	O	O
IL-4	NN	O	B-protein
does	NN	O	O
not	NN	O	O
activate	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
it	NN	O	O
induces	NN	O	O
expression	NN	O	O
of	NN	O	O
STAT5-regulated	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
CTLL-2	NN	O	B-cell_line
,	NN	O	O
i.e	NN	O	O
.	NN	O	O
beta-casein	NN	O	B-protein
,	NN	O	O
a	NN	O	O
cytokine-inducible	NN	O	B-protein
SH2-containing	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
CIS	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
oncostatin	NN	O	B-protein
M	NN	O	I-protein
(	NN	O	O
OSM	NN	O	B-protein
)	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
STAT6	NN	O	O
activated	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
substitutes	NN	O	O
for	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

IL-2	NN	O	B-protein
-induced	NN	O	O
beta-casein	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
enhanced	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
synergistic	NN	O	O
effect	NN	O	O
of	NN	O	O
Dexamethasone	NN	O	O
requires	NN	O	O
the	NN	O	O
sequence	NN	O	O
between	NN	O	O
-155	NN	O	B-DNA
and	NN	O	I-DNA
-193	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
beta-casein	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Coincidentally	NN	O	O
,	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
enhanced	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
beta-casein	NN	O	B-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
an	NN	O	O
active	NN	O	O
form	NN	O	O
of	NN	O	O
Ras	NN	O	B-protein
,	NN	O	O
Ras	NN	O	B-protein
(	NN	O	I-protein
G12V	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
suppressed	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
beta-casein	NN	O	B-protein
and	NN	O	O
OSM	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
negative	NN	O	O
effect	NN	O	O
of	NN	O	O
Ras	NN	O	B-protein
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
region	NN	O	O
between	NN	O	O
-105	NN	O	B-DNA
and	NN	O	I-DNA
-193	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
beta-casein	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
apparent	NN	O	O
contradiction	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
form	NN	O	O
of	NN	O	O
Ras	NN	O	B-protein
,	NN	O	O
RasN17	NN	O	B-protein
,	NN	O	O
also	NN	O	O
inhibited	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
containing	NN	O	O
the	NN	O	O
minimal	NN	O	O
beta-casein	NN	O	B-DNA
STAT5	NN	O	I-DNA
element	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
promoters	NN	O	O
of	NN	O	O
CIS	NN	O	B-protein
and	NN	O	O
OSM	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
Ras	NN	O	B-protein
(	NN	O	I-protein
G12V	NN	O	I-protein
)	NN	O	I-protein
complemented	NN	O	O
signaling	NN	O	O
by	NN	O	O
an	NN	O	O
erythropoietin	NN	O	B-protein
receptor	NN	O	I-protein
mutant	NN	O	I-protein
defective	NN	O	O
in	NN	O	O
Ras	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
augmented	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
beta-casein	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
mutant	NN	O	B-protein
erythropoietin	NN	O	I-protein
receptor	NN	O	I-protein
signaling	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
possible	NN	O	O
role	NN	O	O
of	NN	O	O
Ras	NN	O	B-protein
in	NN	O	O
Stat5	NN	O	B-protein
-mediated	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
collectively	NN	O	O
reveal	NN	O	O
a	NN	O	O
complex	NN	O	O
interaction	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
with	NN	O	O
other	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
and	NN	O	O
illustrate	NN	O	O
that	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
requires	NN	O	O
integration	NN	O	O
of	NN	O	O
opposing	NN	O	O
signals	NN	O	O

-DOCSTART-	O

GATA-3	NN	O	B-protein
represses	NN	O	O
gp91phox	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
eosinophil-committed	NN	O	B-cell_line
HL-60-C15	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
regulatory	NN	O	O
mechanism	NN	O	O
of	NN	O	O
gp91phox	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
eosinophils	NN	O	O
,	NN	O	O
we	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
eosinophil-committed	NN	O	B-cell_line
HL-60-C15	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
gp91phox	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
identified	NN	O	O
a	NN	O	O
negative	NN	O	B-DNA
element	NN	O	I-DNA
from	NN	O	O
bp	NN	O	O
-267	NN	O	B-DNA
to	NN	O	I-DNA
-246	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
gp91phox	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
deletion	NN	O	O
of	NN	O	O
which	NN	O	O
caused	NN	O	O
an	NN	O	O
83	NN	O	O
%	NN	O	O
increase	NN	O	O
in	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Electrophoresis	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
GATA-3	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
the	NN	O	O
GATA	NN	O	B-DNA
consensus	NN	O	I-DNA
site	NN	O	I-DNA
from	NN	O	O
bp	NN	O	O
-256	NN	O	B-DNA
to	NN	O	I-DNA
-250	NN	O	I-DNA
.	NN	O	I-DNA

An	NN	O	O
81	NN	O	O
%	NN	O	O
increment	NN	O	O
in	NN	O	O
promoter	NN	O	O
activity	NN	O	O
was	NN	O	O
obtained	NN	O	O
when	NN	O	O
a	NN	O	O
mutation	NN	O	O
was	NN	O	O
introduced	NN	O	O
in	NN	O	O
the	NN	O	O
GATA-3	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
bp	NN	O	B-DNA
-267	NN	O	I-DNA
to	NN	O	I-DNA
+12	NN	O	I-DNA
construct	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
comparable	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
bp	NN	O	B-DNA
-245	NN	O	I-DNA
to	NN	O	I-DNA
+12	NN	O	I-DNA
construct	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
conclude	NN	O	O
that	NN	O	O
GATA-3	NN	O	B-protein
specifically	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
GATA	NN	O	B-DNA
site	NN	O	I-DNA
negatively	NN	O	O
regulates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
in	NN	O	O
HL-60-C15	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Uncoupling	NN	O	O
activation-dependent	NN	O	O
HS1	NN	O	B-protein
phosphorylation	NN	O	O
from	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
transcriptional	NN	O	O
activation	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
:	NN	O	O
differential	NN	O	O
signaling	NN	O	O
through	NN	O	O
CD3	NN	O	B-protein
and	NN	O	O
the	NN	O	O
costimulatory	NN	O	O
receptors	NN	O	O
CD2	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

CD3	NN	O	B-protein
,	NN	O	O
CD2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CD28	NN	O	B-protein
are	NN	O	O
functionally	NN	O	O
distinct	NN	O	O
receptors	NN	O	O
on	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Engagement	NN	O	O
of	NN	O	O
any	NN	O	O
of	NN	O	O
these	NN	O	O
receptors	NN	O	O
induces	NN	O	O
the	NN	O	O
rapid	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
a	NN	O	O
shared	NN	O	O
group	NN	O	O
of	NN	O	O
intracellular	NN	O	B-protein
signaling	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
including	NN	O	O
Vav	NN	O	B-protein
,	NN	O	O
Cbl	NN	O	B-protein
,	NN	O	O
p85	NN	O	B-protein
phosphoinositide	NN	O	I-protein
3-kinase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
Src	NN	O	B-protein
family	NN	O	I-protein
kinases	NN	O	I-protein
Lck	NN	O	B-protein
and	NN	O	O
Fyn	NN	O	B-protein
.	NN	O	O

Ligation	NN	O	O
of	NN	O	O
CD3	NN	O	B-protein
also	NN	O	O
induces	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
HS1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
75-kDa	NN	O	B-protein
hematopoietic	NN	O	I-protein
cell-specific	NN	O	I-protein
intracellular	NN	O	I-protein
signaling	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
unknown	NN	O	O
function	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
changes	NN	O	O
in	NN	O	O
HS1	NN	O	B-protein
phosphorylation	NN	O	O
after	NN	O	O
differential	NN	O	O
stimulation	NN	O	O
of	NN	O	O
CD3	NN	O	B-protein
,	NN	O	O
CD2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CD28	NN	O	B-protein
to	NN	O	O
elucidate	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
to	NN	O	O
further	NN	O	O
delineate	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
recruited	NN	O	O
by	NN	O	O
these	NN	O	O
receptors	NN	O	O
.	NN	O	O

Unlike	NN	O	O
ligation	NN	O	O
of	NN	O	O
CD3	NN	O	B-protein
,	NN	O	O
stimulation	NN	O	O
with	NN	O	O
anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
or	NN	O	O
CHO	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressing	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
ligands	NN	O	I-protein
CD80	NN	O	B-protein
or	NN	O	O
CD86	NN	O	B-protein
did	NN	O	O
not	NN	O	O
lead	NN	O	O
to	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
HS1	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
no	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
HS1	NN	O	B-protein
was	NN	O	O
induced	NN	O	O
by	NN	O	O
mitogenic	NN	O	B-cell_type
pairs	NN	O	I-cell_type
of	NN	O	O
anti-CD2	NN	O	B-protein
mAbs	NN	O	I-protein
capable	NN	O	O
of	NN	O	O
activating	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NFAT	NN	O	I-protein
(	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Costimulation	NN	O	O
through	NN	O	O
CD28	NN	O	B-protein
and/or	NN	O	O
CD2	NN	O	B-protein
did	NN	O	O
not	NN	O	O
modulate	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
-dependent	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
HS1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
studies	NN	O	O
indicated	NN	O	O
that	NN	O	O
CD3	NN	O	B-protein
-induced	NN	O	O
HSI	NN	O	B-protein
phosphorylation	NN	O	O
was	NN	O	O
dependent	NN	O	O
upon	NN	O	O
both	NN	O	O
the	NN	O	O
Src	NN	O	B-protein
family	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
Lck	NN	O	B-protein
and	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
phosphatase	NN	O	I-protein
CD45	NN	O	B-protein
,	NN	O	O
did	NN	O	O
not	NN	O	O
require	NN	O	O
MEK1	NN	O	B-protein
kinase	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
regulated	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
although	NN	O	O
CD3	NN	O	B-protein
,	NN	O	O
CD28	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CD2	NN	O	O
activate	NN	O	O
many	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
signaling	NN	O	B-protein
molecules	NN	O	I-protein
,	NN	O	O
they	NN	O	O
differed	NN	O	O
in	NN	O	O
their	NN	O	O
capacity	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
HSI	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
activation-dependent	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
HS1	NN	O	B-protein
was	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
NFAT	NN	O	B-protein
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

c-fos	NN	O	O
and	NN	O	O
c-jun	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
cord	NN	O	I-cell_type
and	NN	O	I-cell_type
adult	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
an	NN	O	O
analysis	NN	O	O
by	NN	O	O
Northern	NN	O	O
hybridization	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
AND	NN	O	O
OBJECTIVES	NN	O	O
:	NN	O	O
To	NN	O	O
further	NN	O	O
analyze	NN	O	O
the	NN	O	O
neonatal	NN	O	O
immune	NN	O	O
response	NN	O	O
to	NN	O	O
an	NN	O	O
antigenic	NN	O	O
challenge	NN	O	O
such	NN	O	O
as	NN	O	O
blood	NN	O	O
transfusion	NN	O	O
,	NN	O	O
c-fos	NN	O	O
and	NN	O	O
c-jun	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
were	NN	O	O
analyzed	NN	O	O
in	NN	O	O
twelve	NN	O	O
in-vitro-stimulated	NN	O	O
normal	NN	O	O
cord	NN	O	O
blood	NN	O	O
and	NN	O	O
ten	NN	O	O
in-vitro-stimulated	NN	O	B-cell_type
normal	NN	O	I-cell_type
adult	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
samples	NN	O	I-cell_type
.	NN	O	O

MATERIALS	NN	O	O
AND	NN	O	O
METHODS	NN	O	O
:	NN	O	O
Lymphocyte	NN	O	B-cell_type
samples	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
by	NN	O	O
either	NN	O	O
the	NN	O	O
mitogen	NN	O	B-protein
phytohemagglutinin	NN	O	I-protein
(	NN	O	O
PHA	NN	O	B-protein
)	NN	O	O
or	NN	O	O
the	NN	O	O
monoclonal	NN	O	O
antibody	NN	O	O
alphaCD3	NN	O	B-protein
.	NN	O	O

Proliferation	NN	O	O
rate	NN	O	O
and	NN	O	O
Northern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
were	NN	O	O
employed	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Cord	NN	O	O
lymphocytes	NN	O	O
revealed	NN	O	O
a	NN	O	O
greater	NN	O	O
proliferation	NN	O	O
rate	NN	O	O
with	NN	O	O
PHA	NN	O	B-protein
and	NN	O	O
alphaCD3	NN	O	B-protein
than	NN	O	O
adult	NN	O	O
lymphocytes	NN	O	O
(	NN	O	O
p	NN	O	O
=	NN	O	O
0.0081	NN	O	O
and	NN	O	O
0.0023	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
cord	NN	O	O
and	NN	O	O
adult	NN	O	O
samples	NN	O	O
revealed	NN	O	O
similar	NN	O	O
maximal	NN	O	O
increases	NN	O	O
in	NN	O	O
c-fos	NN	O	B-RNA
(	NN	O	O
99+/-15	NN	O	O
and	NN	O	O
126+/-11	NN	O	O
%	NN	O	O
,	NN	O	O
p	NN	O	O
=	NN	O	O
0.0126	NN	O	O
)	NN	O	O
and	NN	O	O
c-jun	NN	O	B-RNA
(	NN	O	I-RNA
123+/-9	NN	O	I-RNA
and	NN	O	I-RNA
185+/-38	NN	O	I-RNA
%	NN	O	I-RNA
,	NN	O	I-RNA
p	NN	O	I-RNA
=	NN	O	I-RNA
0.0291	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
as	NN	O	O
early	NN	O	O
as	NN	O	O
15	NN	O	O
min	NN	O	O
post-	NN	O	O
alphaCD3	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Adult	NN	O	O
lymphocytes	NN	O	O
showed	NN	O	O
an	NN	O	O
equivalent	NN	O	O
increase	NN	O	O
in	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	B-DNA
(	NN	O	O
140+/-25	NN	O	O
and	NN	O	O
155+/-31	NN	O	O
%	NN	O	O
)	NN	O	O
at	NN	O	O
30	NN	O	O
min	NN	O	O
post-	NN	O	O
PHA	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
while	NN	O	O
cord	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
maximum	NN	O	O
c-fos	NN	O	O
and	NN	O	O
c-jun	NN	O	O
expression	NN	O	O
(	NN	O	O
82+/-6	NN	O	O
and	NN	O	O
142+/-12	NN	O	O
%	NN	O	O
)	NN	O	O
occurred	NN	O	O
at	NN	O	O
15	NN	O	O
min	NN	O	O
post-	NN	O	O
PHA	NN	O	B-protein
stimulation	NN	O	O
(	NN	O	O
c-fos	NN	O	B-DNA
,	NN	O	O
p	NN	O	O
=	NN	O	O
0.0354	NN	O	O
;	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	O
p	NN	O	O
=	NN	O	O
0.0112	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Although	NN	O	O
cord	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
proliferation	NN	O	O
rates	NN	O	O
were	NN	O	O
significantly	NN	O	O
greater	NN	O	O
than	NN	O	O
those	NN	O	O
of	NN	O	O
adult	NN	O	O
lymphocytes	NN	O	O
following	NN	O	O
stimulation	NN	O	O
,	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
,	NN	O	O
as	NN	O	O
analyzed	NN	O	O
by	NN	O	O
c-fos	NN	O	O
and	NN	O	O
c-jun	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
,	NN	O	O
appears	NN	O	O
similar	NN	O	O
in	NN	O	O
both	NN	O	O
cord	NN	O	O
and	NN	O	O
adult	NN	O	O
samples	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
cord	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
exhibits	NN	O	O
an	NN	O	O
adult-type	NN	O	O
profile	NN	O	O
.	NN	O	O

-DOCSTART-	O

Fcgamma	NN	O	B-protein
receptor	NN	O	I-protein
-mediated	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
is	NN	O	O
independent	NN	O	O
of	NN	O	O
Ras	NN	O	B-protein
.	NN	O	O

Receptors	NN	O	O
for	NN	O	O
the	NN	O	O
Fc	NN	O	B-protein
portion	NN	O	I-protein
of	NN	O	O
immunoglobulin	NN	O	B-protein
molecules	NN	O	I-protein
(	NN	O	O
FcR	NN	O	B-protein
)	NN	O	O
present	NN	O	O
on	NN	O	O
leukocyte	NN	O	O
cell	NN	O	O
membranes	NN	O	O
mediate	NN	O	O
a	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
cellular	NN	O	O
responses	NN	O	O
that	NN	O	O
are	NN	O	O
very	NN	O	O
important	NN	O	O
in	NN	O	O
host	NN	O	O
defense	NN	O	O
,	NN	O	O
including	NN	O	O
phagocytosis	NN	O	O
,	NN	O	O
cell	NN	O	O
cytotoxicity	NN	O	O
,	NN	O	O
production	NN	O	O
and	NN	O	O
secretion	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
mediators	NN	O	I-protein
,	NN	O	O
and	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Cross-linking	NN	O	O
of	NN	O	O
FcR	NN	O	B-protein
with	NN	O	O
immune	NN	O	B-protein
complexes	NN	O	I-protein
leads	NN	O	O
,	NN	O	O
first	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
protein-tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
events	NN	O	O
that	NN	O	O
follow	NN	O	O
and	NN	O	O
that	NN	O	O
transduce	NN	O	O
signals	NN	O	O
from	NN	O	O
these	NN	O	O
receptors	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
are	NN	O	O
still	NN	O	O
poorly	NN	O	O
defined	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
from	NN	O	O
Fc	NN	O	B-protein
receptors	NN	O	I-protein
that	NN	O	O
leads	NN	O	O
to	NN	O	O
gene	NN	O	O
activation	NN	O	O
and	NN	O	O
production	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Cross-linking	NN	O	O
of	NN	O	O
FcR	NN	O	B-protein
,	NN	O	O
on	NN	O	O
the	NN	O	O
THP-1	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
by	NN	O	O
immune	NN	O	B-protein
complexes	NN	O	I-protein
resulted	NN	O	O
in	NN	O	O
both	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
and	NN	O	O
interleukin	NN	O	B-protein
1	NN	O	I-protein
production	NN	O	O
.	NN	O	O

These	NN	O	O
responses	NN	O	O
were	NN	O	O
completely	NN	O	O
blocked	NN	O	O
by	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
inhibitors	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
mutants	NN	O	I-protein
of	NN	O	O
Ras	NN	O	B-protein
and	NN	O	O
Raf-1	NN	O	B-protein
,	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
any	NN	O	O
effect	NN	O	O
on	NN	O	O
FcR	NN	O	B-protein
-mediated	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
MAPK	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
was	NN	O	O
not	NN	O	O
used	NN	O	O
by	NN	O	O
these	NN	O	O
receptors	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
MAPK	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
easily	NN	O	O
detected	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
kinase	NN	O	O
assays	NN	O	O
,	NN	O	O
after	NN	O	O
FcR	NN	O	B-protein
cross-linking	NN	O	O
with	NN	O	O
immune	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
the	NN	O	O
specific	NN	O	O
MAPK	NN	O	B-protein
/extracellular	NN	O	O
signal-regulated	NN	O	O
kinase	NN	O	O
kinase	NN	O	O
(	NN	O	O
MAPK	NN	O	O
kinase	NN	O	O
)	NN	O	O
inhibitor	NN	O	O
PD98059	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
MAPK	NN	O	B-protein
activation	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
FcR	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
from	NN	O	O
Fc	NN	O	B-protein
receptors	NN	O	I-protein
leading	NN	O	O
to	NN	O	O
expression	NN	O	O
of	NN	O	O
different	NN	O	O
genes	NN	O	O
important	NN	O	O
to	NN	O	O
leukocyte	NN	O	O
biology	NN	O	O
,	NN	O	O
initiates	NN	O	O
with	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
and	NN	O	O
requires	NN	O	O
MAPK	NN	O	B-protein
activation	NN	O	O
;	NN	O	O
but	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
other	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
FcR	NN	O	B-protein
-mediated	NN	O	O
MAPK	NN	O	B-protein
activation	NN	O	O
does	NN	O	O
not	NN	O	O
involve	NN	O	O
Ras	NN	O	B-protein
and	NN	O	O
Raf	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

BCL-6	NN	O	B-protein
mutations	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
germinal	NN	O	I-cell_type
center	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
evidence	NN	O	O
of	NN	O	O
somatic	NN	O	O
hypermutation	NN	O	O
acting	NN	O	O
outside	NN	O	O
Ig	NN	O	B-DNA
loci	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
antigen-driven	NN	O	O
somatic	NN	O	O
hypermutation	NN	O	O
of	NN	O	O
Ig	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
unknown	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
is	NN	O	O
commonly	NN	O	O
believed	NN	O	O
that	NN	O	O
this	NN	O	O
mechanism	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
loci	NN	O	I-DNA
.	NN	O	O

B	NN	O	B-cell_type
cell	NN	O	I-cell_type
lymphomas	NN	O	I-cell_type
commonly	NN	O	O
display	NN	O	O
multiple	NN	O	O
somatic	NN	O	O
mutations	NN	O	O
clustering	NN	O	O
in	NN	O	O
the	NN	O	O
5'-regulatory	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
BCL-6	NN	O	B-protein
,	NN	O	O
a	NN	O	O
proto-oncogene	NN	O	B-DNA
encoding	NN	O	O
for	NN	O	O
a	NN	O	O
POZ/Zinc	NN	O	B-protein
finger	NN	O	I-protein
transcriptional	NN	O	I-protein
repressor	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
(	NN	O	I-cell_type
GC	NN	O	I-cell_type
)	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
required	NN	O	O
for	NN	O	O
GC	NN	O	O
formation	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
BCL-6	NN	O	B-protein
mutations	NN	O	O
represent	NN	O	O
a	NN	O	O
tumor-associated	NN	O	O
phenomenon	NN	O	O
or	NN	O	O
reflect	NN	O	O
a	NN	O	O
physiologic	NN	O	O
mechanism	NN	O	O
,	NN	O	O
we	NN	O	O
screened	NN	O	O
single	NN	O	O
human	NN	O	B-cell_type
tonsillar	NN	O	I-cell_type
GC	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
mutations	NN	O	O
occurring	NN	O	O
in	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-DNA
5'-noncoding	NN	O	I-DNA
region	NN	O	I-DNA
and	NN	O	O
in	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
variable	NN	O	I-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

Thirty	NN	O	O
percent	NN	O	O
of	NN	O	O
GC	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
naive	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
displayed	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
742	NN	O	B-DNA
bp	NN	O	I-DNA
region	NN	O	I-DNA
analyzed	NN	O	O
within	NN	O	O
the	NN	O	O
first	NN	O	O
intron	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
(	NN	O	O
overall	NN	O	O
frequency	NN	O	O
:	NN	O	O
5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-4	NN	O	O
)	NN	O	O
/bp	NN	O	O
)	NN	O	O
.	NN	O	O

Accordingly	NN	O	O
,	NN	O	O
an	NN	O	O
expanded	NN	O	O
survey	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
malignancies	NN	O	O
showed	NN	O	O
that	NN	O	O
BCL-6	NN	O	B-protein
mutations	NN	O	O
are	NN	O	O
restricted	NN	O	O
to	NN	O	O
B	NN	O	O
cell	NN	O	O
tumors	NN	O	O
displaying	NN	O	O
GC	NN	O	O
or	NN	O	O
post-GC	NN	O	O
phenotype	NN	O	O
and	NN	O	O
carrying	NN	O	O
mutated	NN	O	B-DNA
Ig	NN	O	I-DNA
variable	NN	O	I-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
somatic	NN	O	O
hypermutation	NN	O	O
mechanism	NN	O	O
active	NN	O	O
in	NN	O	O
GC	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
physiologically	NN	O	O
targets	NN	O	O
non-Ig	NN	O	B-DNA
sequences	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
AD1	NN	O	B-protein
and	NN	O	O
AD2	NN	O	B-protein
transactivation	NN	O	O
domains	NN	O	O
of	NN	O	O
E2A	NN	O	B-protein
are	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
antiapoptotic	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
oncoprotein	NN	O	I-protein
E2A-HLF	NN	O	B-protein
.	NN	O	O

The	NN	O	O
chimeric	NN	O	B-protein
oncoprotein	NN	O	I-protein
E2A-HLF	NN	O	B-protein
,	NN	O	O
generated	NN	O	O
by	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
17	NN	O	I-DNA
;	NN	O	I-DNA
19	NN	O	I-DNA
)	NN	O	I-DNA
chromosomal	NN	O	I-DNA
translocation	NN	O	I-DNA
in	NN	O	O
pro-B-cell	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
,	NN	O	O
incorporates	NN	O	O
the	NN	O	O
transactivation	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
E2A	NN	O	B-protein
and	NN	O	O
the	NN	O	O
basic	NN	O	B-protein
leucine	NN	O	I-protein
zipper	NN	O	I-protein
(	NN	O	I-protein
bZIP	NN	O	I-protein
)	NN	O	I-protein
DNA-binding	NN	O	I-protein
and	NN	O	I-protein
protein	NN	O	I-protein
dimerization	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
HLF	NN	O	B-protein
(	NN	O	O
hepatic	NN	O	B-protein
leukemic	NN	O	I-protein
factor	NN	O	I-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
E2A-HLF	NN	O	B-protein
to	NN	O	O
prolong	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
interleukin-3	NN	O	B-cell_line
(	NN	O	I-cell_line
IL-3	NN	O	I-cell_line
)	NN	O	I-cell_line
-dependent	NN	O	I-cell_line
murine	NN	O	I-cell_line
pro-B	NN	O	I-cell_line
cells	NN	O	I-cell_line
after	NN	O	O
IL-3	NN	O	B-protein
withdrawal	NN	O	O
suggests	NN	O	O
that	NN	O	O
it	NN	O	O
disrupts	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
normally	NN	O	O
responsible	NN	O	O
for	NN	O	O
cell	NN	O	O
suicide	NN	O	O
,	NN	O	O
allowing	NN	O	O
the	NN	O	O
cells	NN	O	O
to	NN	O	O
accumulate	NN	O	O
as	NN	O	O
transformed	NN	O	B-cell_type
lymphoblasts	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
structural	NN	O	O
motifs	NN	O	O
that	NN	O	O
contribute	NN	O	O
to	NN	O	O
this	NN	O	O
antiapoptotic	NN	O	O
effect	NN	O	O
,	NN	O	O
we	NN	O	O
constructed	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
E2A-HLF	NN	O	B-protein
mutants	NN	O	I-protein
and	NN	O	O
programmed	NN	O	O
their	NN	O	O
expression	NN	O	O
in	NN	O	O
IL-3-dependent	NN	O	B-cell_line
murine	NN	O	I-cell_line
pro-B	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
FL5.12	NN	O	B-cell_line
line	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
zinc-inducible	NN	O	O
vector	NN	O	O
.	NN	O	O

Neither	NN	O	O
the	NN	O	O
E12	NN	O	B-protein
nor	NN	O	O
the	NN	O	O
E47	NN	O	B-protein
product	NN	O	O
of	NN	O	O
the	NN	O	O
E2A	NN	O	B-DNA
gene	NN	O	I-DNA
nor	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
HLF	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
protect	NN	O	O
the	NN	O	O
cells	NN	O	O
from	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-3	NN	O	B-protein
deprivation	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
different	NN	O	O
combinations	NN	O	O
of	NN	O	O
disabling	NN	O	O
mutations	NN	O	O
within	NN	O	O
the	NN	O	O
HLF	NN	O	B-protein
bZIP	NN	O	I-protein
domain	NN	O	I-protein
had	NN	O	O
little	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
antiapoptotic	NN	O	O
property	NN	O	O
of	NN	O	O
the	NN	O	O
chimeric	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
so	NN	O	O
long	NN	O	O
as	NN	O	O
the	NN	O	O
amino-terminal	NN	O	O
portion	NN	O	O
of	NN	O	O
E2A	NN	O	B-protein
remained	NN	O	O
intact	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
a	NN	O	O
bZIP	NN	O	B-protein
domain	NN	O	I-protein
defective	NN	O	O
in	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
mutants	NN	O	O
retaining	NN	O	O
either	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
transactivation	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
E2A	NN	O	B-protein
were	NN	O	O
able	NN	O	O
to	NN	O	O
extend	NN	O	O
cell	NN	O	O
survival	NN	O	O
after	NN	O	O
growth	NN	O	B-protein
factor	NN	O	I-protein
deprivation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
block	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
imposed	NN	O	O
by	NN	O	O
E2A-HLF	NN	O	B-protein
in	NN	O	O
pro-B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
depends	NN	O	O
critically	NN	O	O
on	NN	O	O
the	NN	O	O
transactivating	NN	O	B-protein
regions	NN	O	I-protein
of	NN	O	O
E2A	NN	O	B-protein
.	NN	O	O

Since	NN	O	O
neither	NN	O	O
DNA	NN	O	O
binding	NN	O	O
nor	NN	O	O
protein	NN	O	O
dimerization	NN	O	O
through	NN	O	O
the	NN	O	O
bZIP	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
HLF	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
this	NN	O	O
effect	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
mechanisms	NN	O	O
whereby	NN	O	O
protein-protein	NN	O	O
interactions	NN	O	O
with	NN	O	O
the	NN	O	O
amino-terminal	NN	O	O
region	NN	O	O
of	NN	O	O
E2A	NN	O	B-protein
allow	NN	O	O
the	NN	O	O
chimera	NN	O	B-protein
to	NN	O	O
act	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
cofactor	NN	O	O
to	NN	O	O
alter	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
regulating	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
machinery	NN	O	O
in	NN	O	O
pro-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
Mn	NN	O	B-protein
SOD	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
without	NN	O	O
inflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
production	NN	O	O
by	NN	O	O
a	NN	O	O
mutant	NN	O	B-protein
endotoxin	NN	O	I-protein
.	NN	O	O

Endotoxin	NN	O	B-protein
selectively	NN	O	O
induces	NN	O	O
monocyte	NN	O	B-protein
Mn	NN	O	I-protein
superoxide	NN	O	I-protein
dismutase	NN	O	I-protein
(	NN	O	O
SOD	NN	O	B-protein
)	NN	O	O
without	NN	O	O
affecting	NN	O	O
levels	NN	O	O
of	NN	O	O
Cu	NN	O	O
,	NN	O	O
Zn	NN	O	B-protein
SOD	NN	O	I-protein
,	NN	O	O
catalase	NN	O	B-protein
,	NN	O	O
or	NN	O	O
glutathione	NN	O	B-protein
peroxidase	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
structure-activity	NN	O	O
relationship	NN	O	O
and	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
endotoxin	NN	O	O
induces	NN	O	O
Mn	NN	O	B-protein
SOD	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
a	NN	O	O
mutant	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
endotoxin	NN	O	O
lacking	NN	O	O
myristoyl	NN	O	O
fatty	NN	O	O
acid	NN	O	O
at	NN	O	O
the	NN	O	O
3	NN	O	O
'	NN	O	O
R-3-hydroxymyristate	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
lipid	NN	O	O
A	NN	O	O
moiety	NN	O	O
retained	NN	O	O
its	NN	O	O
full	NN	O	O
capacity	NN	O	O
to	NN	O	O
coagulate	NN	O	O
Limulus	NN	O	O
amoebocyte	NN	O	O
lysate	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
wild-type	NN	O	O
E.	NN	O	O
coli	NN	O	O
endotoxin	NN	O	O
and	NN	O	O
markedly	NN	O	O
stimulated	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-protein
monocyte	NN	O	I-protein
nuclear	NN	O	I-protein
factor-kappaB	NN	O	I-protein
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
Mn	NN	O	B-RNA
SOD	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
wild-type	NN	O	O
endotoxin	NN	O	O
,	NN	O	O
it	NN	O	O
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
significant	NN	O	O
production	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
and	NN	O	O
macrophage	NN	O	B-protein
inflammatory	NN	O	I-protein
protein-1alpha	NN	O	I-protein
by	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
1	NN	O	O
)	NN	O	O
lipid	NN	O	O
A	NN	O	O
myristoyl	NN	O	O
fatty	NN	O	O
acid	NN	O	O
,	NN	O	O
although	NN	O	O
it	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
production	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
not	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
Mn	NN	O	B-protein
SOD	NN	O	I-protein
,	NN	O	O
2	NN	O	O
)	NN	O	O
endotoxin	NN	O	B-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
Mn	NN	O	B-protein
SOD	NN	O	I-protein
and	NN	O	O
inflammatory	NN	O	O
cytokines	NN	O	B-protein
are	NN	O	O
regulated	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
through	NN	O	O
different	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
,	NN	O	O
and	NN	O	O
3	NN	O	O
)	NN	O	O
failure	NN	O	O
of	NN	O	O
the	NN	O	O
mutant	NN	O	B-protein
endotoxin	NN	O	I-protein
to	NN	O	O
induce	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
production	NN	O	O
is	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
due	NN	O	O
to	NN	O	O
its	NN	O	O
inability	NN	O	O
to	NN	O	O
activate	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
vitellogenin	NN	O	B-DNA
gene	NN	O	I-DNA
B1	NN	O	I-DNA
promoter	NN	O	I-DNA
after	NN	O	O
transfer	NN	O	O
into	NN	O	O
hepatocytes	NN	O	B-cell_type
in	NN	O	O
primary	NN	O	O
cultures	NN	O	O
.	NN	O	O

The	NN	O	O
estrogen-dependent	NN	O	O
and	NN	O	O
tissue-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
Xenopus	NN	O	O
laevis	NN	O	O
vitellogenin	NN	O	B-DNA
gene	NN	O	I-DNA
B1	NN	O	I-DNA
promoter	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
studied	NN	O	O
by	NN	O	O
lipid-mediated	NN	O	O
DNA	NN	O	O
transfer	NN	O	O
into	NN	O	O
Xenopus	NN	O	B-cell_type
hepatocytes	NN	O	I-cell_type
in	NN	O	O
primary	NN	O	O
culture	NN	O	O
.	NN	O	O

Hepatocytes	NN	O	O
achieve	NN	O	O
an	NN	O	O
efficient	NN	O	O
hormonal	NN	O	O
control	NN	O	O
of	NN	O	O
this	NN	O	O
promoter	NN	O	O
through	NN	O	O
a	NN	O	O
functional	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
estrogen	NN	O	B-DNA
responsive	NN	O	I-DNA
elements	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
promoter	NN	O	B-DNA
proximal	NN	O	I-DNA
region	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
high	NN	O	O
density	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
CTF/NF-1	NN	O	B-protein
,	NN	O	O
C/EBP	NN	O	B-protein
and	NN	O	O
HNF3	NN	O	B-protein
.	NN	O	O

DNA	NN	O	O
accessibility	NN	O	O
to	NN	O	O
restriction	NN	O	B-protein
enzymes	NN	O	I-protein
within	NN	O	O
the	NN	O	O
chromosomal	NN	O	O
copy	NN	O	O
of	NN	O	O
the	NN	O	O
vitellogenin	NN	O	B-DNA
gene	NN	O	I-DNA
B1	NN	O	I-DNA
promoter	NN	O	I-DNA
shows	NN	O	O
that	NN	O	O
the	NN	O	O
estrogen	NN	O	B-DNA
responsive	NN	O	I-DNA
unit	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
proximal	NN	O	I-DNA
region	NN	O	I-DNA
are	NN	O	O
sensitive	NN	O	O
to	NN	O	O
digestion	NN	O	O
in	NN	O	O
uninduced	NN	O	O
and	NN	O	O
estrogen-induced	NN	O	B-cell_type
hepatocytes	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
erythrocyte	NN	O	O
nuclei	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
support	NN	O	O
the	NN	O	O
notion	NN	O	O
that	NN	O	O
chromatin	NN	O	B-DNA
configuration	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
interplay	NN	O	O
of	NN	O	O
promoter	NN	O	B-DNA
elements	NN	O	I-DNA
mediate	NN	O	O
proper	NN	O	O
hormone-dependent	NN	O	O
and	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
B1	NN	O	B-DNA
vitellogenin	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
differential	NN	O	O
induction	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
complexes	NN	O	I-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cytokines	NN	O	B-protein
,	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
IL-7	NN	O	B-protein
,	NN	O	O
IL-12	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-15	NN	O	B-protein
are	NN	O	O
key	NN	O	O
regulators	NN	O	O
of	NN	O	O
human	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
T	NN	O	O
and	NN	O	O
NK	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
but	NN	O	O
the	NN	O	O
precise	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
give	NN	O	O
rise	NN	O	O
to	NN	O	O
their	NN	O	O
differential	NN	O	O
activities	NN	O	O
within	NN	O	O
these	NN	O	O
cells	NN	O	O
are	NN	O	O
not	NN	O	O
clear	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
reveal	NN	O	O
that	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	O
STATs	NN	O	B-protein
)	NN	O	O
directly	NN	O	O
mediate	NN	O	O
many	NN	O	O
cytokine	NN	O	B-protein
signals	NN	O	O
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
STATs	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
specific	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
IL-12	NN	O	B-protein
induces	NN	O	O
STAT4	NN	O	B-protein
only	NN	O	O
in	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
primary	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
the	NN	O	O
IL-12	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
induces	NN	O	O
different	NN	O	O
C	NN	O	B-protein
epsilon	NN	O	I-protein
GAS	NN	O	I-protein
DNA-protein	NN	O	I-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
both	NN	O	O
T	NN	O	O
and	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
costimulation	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
or	NN	O	O
IL-12	NN	O	B-protein
does	NN	O	O
not	NN	O	O
alter	NN	O	O
their	NN	O	O
own	NN	O	O
preferential	NN	O	O
GAS	NN	O	B-DNA
-like	NN	O	O
DNA	NN	O	O
binding	NN	O	O
patterns	NN	O	O
when	NN	O	O
C	NN	O	B-DNA
epsilon-	NN	O	I-DNA
,	NN	O	I-DNA
Fc	NN	O	I-DNA
gamma	NN	O	I-DNA
RI-	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
SIE	NN	O	I-DNA
GAS	NN	O	I-DNA
motif	NN	O	I-DNA
containing	NN	O	O
oligonucleotide	NN	O	O
probes	NN	O	O
are	NN	O	O
compared	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
induction	NN	O	O
of	NN	O	O
GAS-like	NN	O	B-protein
DNA-protein	NN	O	I-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
by	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-12	NN	O	B-protein
is	NN	O	O
highly	NN	O	O
selective	NN	O	O
and	NN	O	O
represents	NN	O	O
one	NN	O	O
important	NN	O	O
factor	NN	O	O
in	NN	O	O
determining	NN	O	O
specific	NN	O	O
gene	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
synergistically	NN	O	O
induce	NN	O	O
homo-	NN	O	O
and	NN	O	O
heterodimerized	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
STAT3	NN	O	B-protein
in	NN	O	O
both	NN	O	O
NK	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
their	NN	O	O
reported	NN	O	O
synergism	NN	O	O
in	NN	O	O
modulating	NN	O	O
perforin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	I-protein
-7	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
-15	NN	O	I-protein
induce	NN	O	O
multiple	NN	O	B-protein
STAT	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
including	NN	O	O
STAT5a	NN	O	B-protein
,	NN	O	O
STAT5b	NN	O	B-protein
,	NN	O	O
STAT1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
STAT3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
another	NN	O	O
unidentified	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RI	NN	O	I-protein
GAS	NN	O	I-protein
DNA-binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
activated	NN	O	O
STAT5a	NN	O	B-protein
and	NN	O	O
STAT5b	NN	O	B-protein
proteins	NN	O	O
form	NN	O	O
distinct	NN	O	O
Fc	NN	O	O
gamma	NN	O	O
RI	NN	O	O
GAS	NN	O	O
binding	NN	O	O
patterns	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
they	NN	O	O
might	NN	O	O
have	NN	O	O
different	NN	O	O
roles	NN	O	O
in	NN	O	O
gene	NN	O	O
regulation	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
differential	NN	O	O
responses	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
cell	NN	O	O
activation	NN	O	O
seen	NN	O	O
in	NN	O	O
primary	NN	O	O
NK	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
on	NN	O	O
direct	NN	O	O
stimulation	NN	O	O
with	NN	O	O
different	NN	O	O
cytokines	NN	O	B-protein
may	NN	O	O
be	NN	O	O
a	NN	O	O
direct	NN	O	O
result	NN	O	O
of	NN	O	O
distinct	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
CD95	NN	O	B-DNA
(	NN	O	I-DNA
APO-1/Fas	NN	O	I-DNA
)	NN	O	I-DNA
ligand	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
CD95	NN	O	B-protein
(	NN	O	I-protein
APO-1/Fas	NN	O	I-protein
)	NN	O	I-protein
ligand	NN	O	I-protein
(	NN	O	O
CD95L	NN	O	B-protein
)	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
a	NN	O	O
major	NN	O	O
cause	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
activation-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
CD95L	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CD95L	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Deletion	NN	O	O
studies	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
CD95L	NN	O	B-DNA
proximal	NN	O	I-DNA
promoter	NN	O	I-DNA
sequence	NN	O	I-DNA
from	NN	O	O
-220	NN	O	B-DNA
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
stimulation-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
CD95L	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
discovered	NN	O	O
a	NN	O	O
novel	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
located	NN	O	O
at	NN	O	O
-120	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
CD95L	NN	O	B-DNA
promoter	NN	O	I-DNA
which	NN	O	O
contains	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
SP-1	NN	O	B-protein
and	NN	O	O
a	NN	O	O
yet	NN	O	O
unknown	NN	O	O
inducible	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

Mutation	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	O
factor	NN	O	O
to	NN	O	O
the	NN	O	O
-120	NN	O	B-DNA
region	NN	O	I-DNA
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
the	NN	O	O
biological	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
CD95L	NN	O	B-DNA
promoter	NN	O	I-DNA
upon	NN	O	O
T	NN	O	O
cell	NN	O	O
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
at	NN	O	O
-120	NN	O	B-DNA
also	NN	O	O
contains	NN	O	O
two	NN	O	O
DNA	NN	O	B-DNA
motifs	NN	O	I-DNA
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

NF-AT	NN	O	B-protein
does	NN	O	O
not	NN	O	O
directly	NN	O	O
bind	NN	O	O
to	NN	O	O
this	NN	O	O
element	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
cotransfection	NN	O	O
studies	NN	O	O
with	NN	O	O
an	NN	O	O
NF-AT	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
showed	NN	O	O
that	NN	O	O
NF-AT	NN	O	B-protein
may	NN	O	O
confer	NN	O	O
a	NN	O	O
strong	NN	O	O
inducible	NN	O	O
activity	NN	O	O
to	NN	O	O
the	NN	O	O
CD95L	NN	O	B-DNA
promoter	NN	O	I-DNA
at	NN	O	O
this	NN	O	O
regulatory	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
data	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
agent	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
down-regulates	NN	O	O
CD95L	NN	O	B-protein
transcription	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
this	NN	O	O
positive	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA

-DOCSTART-	O

A	NN	O	O
nongenomic	NN	O	O
mechanism	NN	O	O
for	NN	O	O
progesterone-mediated	NN	O	O
immunosuppression	NN	O	O
:	NN	O	O
inhibition	NN	O	O
of	NN	O	O
K+	NN	O	B-protein
channels	NN	O	I-protein
,	NN	O	O
Ca2+	NN	O	O
signaling	NN	O	O
,	NN	O	O
and	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
progesterone	NN	O	O
causes	NN	O	O
localized	NN	O	O
suppression	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
during	NN	O	O
pregnancy	NN	O	O
has	NN	O	O
remained	NN	O	O
elusive	NN	O	O
.	NN	O	O

Using	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
progesterone	NN	O	O
,	NN	O	O
at	NN	O	O
concentrations	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
placenta	NN	O	O
,	NN	O	O
rapidly	NN	O	O
and	NN	O	O
reversibly	NN	O	O
blocks	NN	O	O
voltage-gated	NN	O	B-protein
and	NN	O	I-protein
calcium-activated	NN	O	I-protein
K+	NN	O	I-protein
channels	NN	O	I-protein
(	NN	O	O
KV	NN	O	B-protein
and	NN	O	O
KCa	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
depolarization	NN	O	O
of	NN	O	O
the	NN	O	O
membrane	NN	O	O
potential	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
result	NN	O	O
,	NN	O	O
Ca2+	NN	O	O
signaling	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
-driven	NN	O	O
gene	NN	O	O
expression	NN	O	O
are	NN	O	O
inhibited	NN	O	O
.	NN	O	O

Progesterone	NN	O	O
acts	NN	O	O
distally	NN	O	O
to	NN	O	O
the	NN	O	O
initial	NN	O	O
steps	NN	O	O
of	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
,	NN	O	O
since	NN	O	O
it	NN	O	O
blocks	NN	O	O
sustained	NN	O	O
Ca2+	NN	O	O
signals	NN	O	O
after	NN	O	O
thapsigargin	NN	O	O
stimulation	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
oscillatory	NN	O	O
Ca2+	NN	O	O
signals	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
Ca2+	NN	O	O
transient	NN	O	O
after	NN	O	O
TCR	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

K+	NN	O	O
channel	NN	O	O
blockade	NN	O	O
by	NN	O	O
progesterone	NN	O	O
is	NN	O	O
specific	NN	O	O
;	NN	O	O
other	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
had	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
progesterone	NN	O	O
antagonist	NN	O	O
RU	NN	O	O
486	NN	O	O
also	NN	O	O
blocked	NN	O	O
KV	NN	O	B-protein
and	NN	O	O
KCa	NN	O	B-protein
channels	NN	O	O
.	NN	O	O

Progesterone	NN	O	O
effectively	NN	O	O
blocked	NN	O	O
a	NN	O	O
broad	NN	O	O
spectrum	NN	O	O
of	NN	O	O
K+	NN	O	B-protein
channels	NN	O	I-protein
,	NN	O	O
reducing	NN	O	O
both	NN	O	O
Kv1.3	NN	O	B-protein
and	NN	O	O
charybdotoxin-resistant	NN	O	B-protein
components	NN	O	I-protein
of	NN	O	O
KV	NN	O	B-protein
current	NN	O	O
and	NN	O	O
KCa	NN	O	B-protein
current	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
blocking	NN	O	O
several	NN	O	O
cloned	NN	O	B-protein
KV	NN	O	I-protein
channels	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Progesterone	NN	O	O
had	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
a	NN	O	O
cloned	NN	O	B-protein
voltage-gated	NN	O	I-protein
Na+	NN	O	I-protein
channel	NN	O	I-protein
,	NN	O	O
an	NN	O	O
inward	NN	O	O
rectifier	NN	O	O
K+	NN	O	B-protein
channel	NN	O	I-protein
,	NN	O	O
or	NN	O	O
on	NN	O	O
lymphocyte	NN	O	B-protein
Ca2+	NN	O	I-protein
and	NN	O	I-protein
Cl-	NN	O	I-protein
channels	NN	O	I-protein
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
direct	NN	O	O
inhibition	NN	O	O
of	NN	O	O
K+	NN	O	B-protein
channels	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
progesterone	NN	O	O
contributes	NN	O	O
to	NN	O	O
progesterone-induced	NN	O	O
immunosuppression	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
mouse	NN	O	O
carrying	NN	O	O
genetic	NN	O	O
defect	NN	O	O
in	NN	O	O
the	NN	O	O
choice	NN	O	O
between	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Transgenic	NN	O	O
mice	NN	O	O
with	NN	O	O
human	NN	O	B-DNA
CD3epsilon	NN	O	I-DNA
gene	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
exhibit	NN	O	O
early	NN	O	O
arrest	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
,	NN	O	O
instead	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
generated	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
of	NN	O	O
a	NN	O	O
line	NN	O	O
,	NN	O	O
tg	NN	O	O
epsilon26	NN	O	O
,	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
CD3epsilon	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
accumulation	NN	O	O
of	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
was	NN	O	O
found	NN	O	O
only	NN	O	O
in	NN	O	O
tg	NN	O	O
epsilon26	NN	O	O
mice	NN	O	O
,	NN	O	O
not	NN	O	O
in	NN	O	O
other	NN	O	O
human	NN	O	B-cell_line
CD3epsilon	NN	O	I-cell_line
transgenic	NN	O	I-cell_line
mouse	NN	O	I-cell_line
lines	NN	O	I-cell_line
or	NN	O	O
other	NN	O	O
T	NN	O	O
cell-deficient	NN	O	O
mice	NN	O	O
,	NN	O	O
including	NN	O	O
CD3-epsilon	NN	O	O
knockout	NN	O	O
mice	NN	O	O
and	NN	O	O
TCR-beta/TCR-delta	NN	O	O
double	NN	O	O
knockout	NN	O	O
mice	NN	O	O
.	NN	O	O

Hanging	NN	O	O
drop-mediated	NN	O	O
transfer	NN	O	O
into	NN	O	O
2-deoxyguanosine-treated	NN	O	O
thymus	NN	O	O
lobes	NN	O	O
showed	NN	O	O
that	NN	O	O
lymphoid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
rather	NN	O	O
than	NN	O	O
thymus	NN	O	O
stromal	NN	O	O
cells	NN	O	O
were	NN	O	O
responsible	NN	O	O
for	NN	O	O
abnormal	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
in	NN	O	O
tg	NN	O	B-protein
epsilon26	NN	O	I-protein
thymus	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
tg	NN	O	B-cell_type
epsilon26	NN	O	I-cell_type
fetal	NN	O	I-cell_type
liver	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
destined	NN	O	O
to	NN	O	O
become	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
normal	NN	O	O
thymus	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
undergoing	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
lymphoid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
tg	NN	O	B-protein
epsilon26	NN	O	I-protein
mice	NN	O	O
are	NN	O	O
genetically	NN	O	O
defective	NN	O	O
in	NN	O	O
thymic	NN	O	O
choice	NN	O	O
between	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
generating	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
even	NN	O	O
in	NN	O	O
normal	NN	O	O
thymus	NN	O	O
environment	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
tg	NN	O	B-cell_type
epsilon26	NN	O	I-cell_type
thymocytes	NN	O	I-cell_type
expressed	NN	O	O
GATA-3	NN	O	B-protein
and	NN	O	O
TCF-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
LEF-1	NN	O	B-protein
and	NN	O	O
PEBP-2alpha	NN	O	B-protein
,	NN	O	O
among	NN	O	O
T	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
early	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
GATA-3	NN	O	B-protein
and	NN	O	O
TCF-1	NN	O	B-protein
expressed	NN	O	O
during	NN	O	O
thymocyte	NN	O	O
development	NN	O	O
do	NN	O	O
not	NN	O	O
necessarily	NN	O	O
determine	NN	O	O
the	NN	O	O
cell	NN	O	O
fate	NN	O	O
into	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
tg	NN	O	B-protein
epsilon26	NN	O	I-protein
mice	NN	O	O
provide	NN	O	O
a	NN	O	O
novel	NN	O	O
mouse	NN	O	O
model	NN	O	O
in	NN	O	O
that	NN	O	O
lineage	NN	O	O
choice	NN	O	O
between	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
genetically	NN	O	O
defective	NN	O	O
.	NN	O	O

-DOCSTART-	O

Downstream	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
TATA-less	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
Bob1	NN	O	O
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
directs	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
homing	NN	O	B-protein
receptor	NN	O	I-protein
BLR1	NN	O	I-protein
to	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
chemokine	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
BLR1	NN	O	B-protein
,	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
regulator	NN	O	O
of	NN	O	O
the	NN	O	O
microenvironmental	NN	O	O
homing	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
lymphoid	NN	O	O
organs	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
studies	NN	O	O
identify	NN	O	O
three	NN	O	O
essential	NN	O	O
elements	NN	O	O
of	NN	O	O
the	NN	O	O
TATA-less	NN	O	B-DNA
blr1	NN	O	I-DNA
core	NN	O	I-DNA
promoter	NN	O	I-DNA
that	NN	O	O
confer	NN	O	O
cell	NN	O	O
type-	NN	O	O
and	NN	O	O
differentiation-specific	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
both	NN	O	O
humans	NN	O	O
and	NN	O	O
mice	NN	O	O
,	NN	O	O
a	NN	O	O
functional	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
-36	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
a	NN	O	O
NF-kappaB	NN	O	B-DNA
motif	NN	O	I-DNA
(	NN	O	O
+44	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
noncanonical	NN	O	B-DNA
octamer	NN	O	I-DNA
motif	NN	O	I-DNA
(	NN	O	O
+157	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
importance	NN	O	O
of	NN	O	O
these	NN	O	O
sites	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
in	NN	O	O
vivo	NN	O	O
studies	NN	O	O
in	NN	O	O
gene-targeted	NN	O	O
mice	NN	O	O
deficient	NN	O	O
of	NN	O	O
either	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
Bob1	NN	O	B-protein
,	NN	O	O
or	NN	O	O
both	NN	O	O
NF-kappaB	NN	O	B-protein
subunits	NN	O	O
p50	NN	O	O
and	NN	O	O
p52	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
of	NN	O	O
these	NN	O	O
animals	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
BLR1	NN	O	B-protein
was	NN	O	O
reduced	NN	O	O
or	NN	O	O
absent	NN	O	O
.	NN	O	O

In	NN	O	O
mice	NN	O	O
deficient	NN	O	O
only	NN	O	O
of	NN	O	O
p52/NF-kappaB	NN	O	B-protein
,	NN	O	O
BLR1	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
unaffected	NN	O	O
.	NN	O	O

Thus	NN	O	O
our	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
BLR1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
target	NN	O	O
gene	NN	O	O
for	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
Bob1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
family	NN	O	I-protein
and	NN	O	O
provides	NN	O	O
a	NN	O	O
link	NN	O	O
to	NN	O	O
the	NN	O	O
impaired	NN	O	O
B	NN	O	O
cell	NN	O	O
functions	NN	O	O
in	NN	O	O
mice	NN	O	O
deficient	NN	O	O
for	NN	O	O
these	NN	O	O
factors	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
transactivation	NN	O	O
by	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
6	NN	O	O
.	NN	O	O

The	NN	O	O
observation	NN	O	O
that	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
6	NN	O	O
(	NN	O	O
HHV-6	NN	O	O
)	NN	O	O
can	NN	O	O
induce	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
and	NN	O	O
expression	NN	O	O
in	NN	O	O
CD4	NN	O	B-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
reported	NN	O	O
several	NN	O	O
years	NN	O	O
ago	NN	O	O
(	NN	O	O
P.Lusso	NN	O	O
,	NN	O	O
A.De	NN	O	O
Maria	NN	O	O
,	NN	O	O
M.Malnati	NN	O	O
,	NN	O	O
F.Lori	NN	O	O
,	NN	O	O
S.E.DeRocco	NN	O	O
,	NN	O	O
M.	NN	O	O
Baseler	NN	O	O
,	NN	O	O
and	NN	O	O
R.C.Gallo	NN	O	O
,	NN	O	O
Nature	NN	O	O
349	NN	O	O
:	NN	O	O
533-535	NN	O	O
,	NN	O	O
1991	NN	O	O
)	NN	O	O
and	NN	O	O
subsequently	NN	O	O
confirmed	NN	O	O
(	NN	O	O
P.Lusso	NN	O	O
,	NN	O	O
M.S.Malnati	NN	O	O
,	NN	O	O
A.Garzino-Demo	NN	O	O
,	NN	O	O
R.W.Crowley	NN	O	O
,	NN	O	O
E.	NN	O	O
O.Long	NN	O	O
,	NN	O	O
and	NN	O	O
R.C.Gallo	NN	O	O
,	NN	O	O
Nature	NN	O	O
362	NN	O	O
:	NN	O	O
458-462	NN	O	O
,	NN	O	O
1993	NN	O	O
;	NN	O	O
G.Furlini	NN	O	O
,	NN	O	O
M.	NN	O	O
Vignoli	NN	O	O
,	NN	O	O
E.Ramazzotti	NN	O	O
,	NN	O	O
M.C.Re	NN	O	O
,	NN	O	O
G.Visani	NN	O	O
,	NN	O	O
and	NN	O	O
M.LaPlaca	NN	O	O
,	NN	O	O
Blood	NN	O	O
87	NN	O	O
:	NN	O	O
4737-4745	NN	O	O
,	NN	O	O
1996	NN	O	O
)	NN	O	O
.	NN	O	O

Our	NN	O	O
objective	NN	O	O
was	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
such	NN	O	O
phenomena	NN	O	O
.	NN	O	O

Using	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
driven	NN	O	O
by	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
HHV-6	NN	O	O
can	NN	O	O
efficiently	NN	O	O
transactivate	NN	O	O
such	NN	O	O
genetic	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	O
DNA	NN	O	O
polymerase	NN	O	O
inhibitor	NN	O	O
phosphonoformic	NN	O	O
acid	NN	O	O
,	NN	O	O
which	NN	O	O
limits	NN	O	O
expression	NN	O	O
to	NN	O	O
the	NN	O	O
immediate-early	NN	O	B-DNA
and	NN	O	I-DNA
early	NN	O	I-DNA
classes	NN	O	I-DNA
of	NN	O	I-DNA
viral	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
deletion	NN	O	O
mutants	NN	O	O
and	NN	O	O
specific	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
mutants	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
identified	NN	O	O
an	NN	O	O
ATF/CRE	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
located	NN	O	O
at	NN	O	O
nucleotides	NN	O	B-DNA
-67	NN	O	I-DNA
to	NN	O	I-DNA
-60	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
HHV-6	NN	O	O
transactivation	NN	O	O
.	NN	O	O

The	NN	O	O
ATF/CRE	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
also	NN	O	O
essential	NN	O	O
for	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
activation	NN	O	O
by	NN	O	O
forskolin	NN	O	O
,	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
adenylate	NN	O	O
cyclase	NN	O	O
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
and	NN	O	O
specific	NN	O	O
antibodies	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
CREB-1	NN	O	B-protein
binds	NN	O	O
specifically	NN	O	O
to	NN	O	O
the	NN	O	O
-79	NN	O	B-DNA
to	NN	O	I-DNA
-52	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Last	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
two	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frames	NN	O	I-DNA
(	NN	O	O
ORFs	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
HHV-6	NN	O	O
,	NN	O	O
U86	NN	O	B-DNA
and	NN	O	O
U89	NN	O	B-DNA
from	NN	O	O
the	NN	O	O
immediate-early	NN	O	O
locus	NN	O	O
A	NN	O	O
,	NN	O	O
that	NN	O	O
can	NN	O	O
transactivate	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
ORFs	NN	O	B-DNA
U86	NN	O	B-DNA
and	NN	O	O
U89	NN	O	B-DNA
is	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
additional	NN	O	O
HHV-6	NN	O	B-DNA
ORFs	NN	O	I-DNA
are	NN	O	O
likely	NN	O	O
to	NN	O	O
contribute	NN	O	O
to	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
will	NN	O	O
help	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
complex	NN	O	O
interactions	NN	O	O
occurring	NN	O	O
between	NN	O	O
HHV-6	NN	O	O
and	NN	O	O
the	NN	O	O
CD4	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
provide	NN	O	O
additional	NN	O	O
information	NN	O	O
regarding	NN	O	O
the	NN	O	O
class	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
CD4	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
PEBP2betaMYH11	NN	O	B-protein
fusion	NN	O	O
created	NN	O	O
by	NN	O	O
Inv	NN	O	B-DNA
(	NN	O	I-DNA
16	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
p13	NN	O	I-DNA
;	NN	O	I-DNA
q22	NN	O	I-DNA
)	NN	O	I-DNA
in	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
impairs	NN	O	O
neutrophil	NN	O	O
maturation	NN	O	O
and	NN	O	O
contributes	NN	O	O
to	NN	O	O
granulocytic	NN	O	O
dysplasia	NN	O	O
.	NN	O	O

Chromosomal	NN	O	O
translocations	NN	O	O
involving	NN	O	O
the	NN	O	O
genes	NN	O	O
encoding	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
and	NN	O	I-protein
beta	NN	O	I-protein
subunits	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Pebp2/Cbf	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
have	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
human	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
and	NN	O	O
the	NN	O	O
preleukemic	NN	O	O
condition	NN	O	O
,	NN	O	O
myelodysplasia	NN	O	O
.	NN	O	O

Inv	NN	O	B-DNA
(	NN	O	I-DNA
16	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
p13	NN	O	I-DNA
;	NN	O	I-DNA
q22	NN	O	I-DNA
)	NN	O	I-DNA
fuses	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
beta	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
Pebp2	NN	O	B-protein
to	NN	O	O
the	NN	O	O
MYH11	NN	O	B-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
a	NN	O	O
smooth	NN	O	B-protein
muscle	NN	O	I-protein
myosin	NN	O	I-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
Smmhc	NN	O	B-protein
)	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
inv	NN	O	B-DNA
(	NN	O	I-DNA
16	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
p13	NN	O	I-DNA
;	NN	O	I-DNA
q22	NN	O	I-DNA
)	NN	O	I-DNA
on	NN	O	O
myelopoiesis	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
the	NN	O	O
hMRP8	NN	O	B-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
to	NN	O	O
generate	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
expressing	NN	O	O
the	NN	O	O
Pebp2beta	NN	O	B-protein
Smmhc	NN	O	B-protein
chimeric	NN	O	I-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Neutrophil	NN	O	O
maturation	NN	O	O
was	NN	O	O
impaired	NN	O	O
in	NN	O	O
PEBP2betaMYH11	NN	O	B-protein
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
had	NN	O	O
normal	NN	O	O
numbers	NN	O	O
of	NN	O	O
circulating	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
,	NN	O	O
their	NN	O	O
bone	NN	O	O
marrow	NN	O	O
contained	NN	O	O
increased	NN	O	O
numbers	NN	O	O
of	NN	O	O
immature	NN	O	B-cell_type
neutrophilic	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
exhibited	NN	O	O
abnormal	NN	O	O
characteristics	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
PEBP2betaMYH11	NN	O	B-protein
inhibited	NN	O	O
neutrophilic	NN	O	O
differentiation	NN	O	O
in	NN	O	O
colonies	NN	O	O
derived	NN	O	O
from	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

Coexpression	NN	O	O
of	NN	O	O
both	NN	O	O
PEBP2betaMYH11	NN	O	B-protein
and	NN	O	O
activated	NN	O	B-protein
NRAS	NN	O	I-protein
induced	NN	O	O
a	NN	O	O
more	NN	O	O
severe	NN	O	O
phenotype	NN	O	O
characterized	NN	O	O
by	NN	O	O
abnormal	NN	O	O
nuclear	NN	O	O
morphology	NN	O	O
indicative	NN	O	O
of	NN	O	O
granulocytic	NN	O	O
dysplasia	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
PEBP2betaMYH11	NN	O	B-protein
can	NN	O	O
impair	NN	O	O
neutrophil	NN	O	O
development	NN	O	O
and	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
alterations	NN	O	O
of	NN	O	O
Pebp2	NN	O	B-protein
can	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
genesis	NN	O	O
of	NN	O	O
myelodysplasia	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
virus-I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
Tax	NN	O	B-protein
-mediated	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
an	NN	O	O
enhanced	NN	O	O
intracellular	NN	O	O
prooxidant	NN	O	O
state	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
an	NN	O	O
estradiol-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
virus-I	NN	O	O
Tax	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
a	NN	O	O
hormone-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
promotes	NN	O	O
an	NN	O	O
enhanced	NN	O	O
prooxidant	NN	O	O
state	NN	O	O
in	NN	O	O
stably	NN	O	B-cell_line
transfected	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
measured	NN	O	O
by	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
intracellular	NN	O	O
levels	NN	O	O
of	NN	O	O
glutathione	NN	O	O
and	NN	O	O
H2O2	NN	O	O
;	NN	O	O
these	NN	O	O
changes	NN	O	O
are	NN	O	O
followed	NN	O	O
by	NN	O	O
apoptotic	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Additional	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
/TCR	NN	O	B-protein
pathway	NN	O	O
enhances	NN	O	O
the	NN	O	O
oxidative	NN	O	O
and	NN	O	O
apoptotic	NN	O	O
effects	NN	O	O
.	NN	O	O

Both	NN	O	O
Tax	NN	O	B-protein
-mediated	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
can	NN	O	O
be	NN	O	O
potently	NN	O	O
suppressed	NN	O	O
by	NN	O	O
antioxidants	NN	O	O
,	NN	O	O
as	NN	O	O
is	NN	O	O
seen	NN	O	O
with	NN	O	O
the	NN	O	O
administration	NN	O	O
of	NN	O	O
recombinant	NN	O	B-protein
thioredoxin	NN	O	I-protein
(	NN	O	O
adult	NN	O	B-protein
T	NN	O	I-protein
cell	NN	O	I-protein
leukemia-derived	NN	O	I-protein
factor	NN	O	I-protein
)	NN	O	O
or	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
.	NN	O	O

Hormone-induced	NN	O	O
Tax	NN	O	B-protein
activation	NN	O	O
induces	NN	O	O
a	NN	O	O
long-lasting	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
target	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
.	NN	O	O

The	NN	O	O
long-term	NN	O	O
exposure	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
hormone	NN	O	O
eventually	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
selection	NN	O	O
of	NN	O	O
cell	NN	O	B-cell_line
clones	NN	O	I-cell_line
that	NN	O	O
have	NN	O	O
lost	NN	O	O
Tax	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

A	NN	O	O
subsequent	NN	O	O
transfection	NN	O	O
of	NN	O	O
these	NN	O	O
apparently	NN	O	O
``	NN	O	B-cell_line
nonresponsive	NN	O	I-cell_line
''	NN	O	I-cell_line
clones	NN	O	I-cell_line
allows	NN	O	O
the	NN	O	O
recovery	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
responses	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Our	NN	O	O
observations	NN	O	O
indicate	NN	O	O
that	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
intracellular	NN	O	O
redox	NN	O	O
status	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
determining	NN	O	O
factor	NN	O	O
in	NN	O	O
Tax	NN	O	B-protein
-mediated	NN	O	O
DNA	NN	O	O
damage	NN	O	O
,	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
and	NN	O	O
selection	NN	O	O
against	NN	O	O
the	NN	O	O
long-term	NN	O	O
expression	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
allosteric	NN	O	O
drug	NN	O	O
,	NN	O	O
o	NN	O	O
,	NN	O	O
o'-bismyristoyl	NN	O	O
thiamine	NN	O	O
disulfide	NN	O	O
,	NN	O	O
suppresses	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
through	NN	O	O
prevention	NN	O	O
of	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
both	NN	O	O
HIV-1	NN	O	B-protein
Tat	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
efficacy	NN	O	O
of	NN	O	O
o	NN	O	O
,	NN	O	O
o'-bismyristoyl	NN	O	O
thiamine	NN	O	O
disulfide	NN	O	O
(	NN	O	O
BMT	NN	O	O
)	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
detail	NN	O	O
against	NN	O	O
HIV-1	NN	O	O
laboratory	NN	O	O
isolates	NN	O	O
(	NN	O	O
HTLV-IIIB	NN	O	O
,	NN	O	O
JRFL	NN	O	O
,	NN	O	O
and	NN	O	O
MN	NN	O	O
)	NN	O	O
,	NN	O	O
primary	NN	O	O
isolates	NN	O	O
(	NN	O	O
KMT	NN	O	O
and	NN	O	O
KMO	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
simian	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
SIVmac251	NN	O	O
)	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

BMT	NN	O	O
inhibited	NN	O	O
the	NN	O	O
replication	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
both	NN	O	O
laboratory	NN	O	O
and	NN	O	O
primary	NN	O	O
isolates	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
BMT	NN	O	O
exhibited	NN	O	O
antiviral	NN	O	O
activity	NN	O	O
against	NN	O	O
SIVmac251	NN	O	O
.	NN	O	O

Minimizing	NN	O	O
energy	NN	O	O
studies	NN	O	O
of	NN	O	O
BMT	NN	O	O
structure	NN	O	O
reveal	NN	O	O
that	NN	O	O
a	NN	O	O
trans-disulfide	NN	O	O
of	NN	O	O
thiamine	NN	O	O
(	NN	O	O
holo	NN	O	O
drug	NN	O	O
)	NN	O	O
disulfide	NN	O	O
(	NN	O	O
TDS	NN	O	O
,	NN	O	O
protodrug	NN	O	O
)	NN	O	O
is	NN	O	O
allosterically	NN	O	O
transited	NN	O	O
to	NN	O	O
the	NN	O	O
reactive	NN	O	O
twisted	NN	O	O
disulfide	NN	O	O
of	NN	O	O
BMT	NN	O	O
(	NN	O	O
allo	NN	O	O
drug	NN	O	O
)	NN	O	O
by	NN	O	O
o	NN	O	O
,	NN	O	O
o'-bismyristoyl	NN	O	O
esterification	NN	O	O
of	NN	O	O
TDS	NN	O	O
.	NN	O	O

BMT	NN	O	O
inhibits	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
both	NN	O	O
HIV-1	NN	O	B-protein
transactivator	NN	O	I-protein
(	NN	O	O
TAT	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
transcriptional	NN	O	I-protein
nuclear	NN	O	I-protein
factor-KB	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid-induced	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Glucocorticoid-induced	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
glucocorticoid-sensitive	NN	O	B-cell_line
6TG1.1	NN	O	I-cell_line
and	NN	O	O
resistant	NN	O	B-cell_line
ICR27TK.3	NN	O	I-cell_line
human	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Following	NN	O	O
glucocorticoid	NN	O	O
treatment	NN	O	O
of	NN	O	O
6TG1.1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
chromatin	NN	O	B-DNA
fragmentation	NN	O	O
was	NN	O	O
observed	NN	O	O
after	NN	O	O
a	NN	O	O
delay	NN	O	O
of	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

Fragmentation	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
ICR27TK.3	NN	O	B-cell_line
cells	NN	O	I-cell_line
containing	NN	O	O
mutant	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
L753F	NN	O	B-cell_line
)	NN	O	O
that	NN	O	O
are	NN	O	O
activation-deficient	NN	O	O
but	NN	O	O
retain	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
repress	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Nor	NN	O	O
was	NN	O	O
fragmentation	NN	O	O
observed	NN	O	O
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
RU38486	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
repression	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
not	NN	O	O
involved	NN	O	O
.	NN	O	O

As	NN	O	O
described	NN	O	O
in	NN	O	O
other	NN	O	O
systems	NN	O	O
,	NN	O	O
fragmentation	NN	O	O
required	NN	O	O
ongoing	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
with	NN	O	O
cycloheximide	NN	O	O
anytime	NN	O	O
during	NN	O	O
the	NN	O	O
first	NN	O	O
18	NN	O	O
h	NN	O	O
of	NN	O	O
steroid	NN	O	O
treatment	NN	O	O
was	NN	O	O
as	NN	O	O
effective	NN	O	O
in	NN	O	O
blocking	NN	O	O
chromatin	NN	O	O
fragmentation	NN	O	O
as	NN	O	O
inhibition	NN	O	O
for	NN	O	O
the	NN	O	O
entire	NN	O	O
period	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
component	NN	O	O
with	NN	O	O
a	NN	O	O
rapid	NN	O	O
turnover	NN	O	O
rate	NN	O	O
is	NN	O	O
required	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
treatment	NN	O	O
completely	NN	O	O
blocked	NN	O	O
12-O-tetradecanoylphorbol	NN	O	O
13-acetate	NN	O	O
induction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
and	NN	O	O
elicited	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
immunoreactive	NN	O	B-protein
IkappaB	NN	O	I-protein
alpha	NN	O	I-protein
in	NN	O	O
sensitive	NN	O	O
6TG1.1	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
resistant	NN	O	O
ICR27TK.3	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
mild	NN	O	O
detergent	NN	O	O
treatment	NN	O	O
of	NN	O	O
cell	NN	O	O
extracts	NN	O	O
indicated	NN	O	O
that	NN	O	O
a	NN	O	O
substantial	NN	O	O
amount	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	B-protein
NF-kappaB	NN	O	I-protein
is	NN	O	O
complexed	NN	O	O
with	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
or	NN	O	O
some	NN	O	O
other	NN	O	O
inhibitory	NN	O	O
factor.	NN	O	O
These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
labile	NN	O	B-protein
inhibitory	NN	O	I-protein
factor	NN	O	I-protein
such	NN	O	O
as	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
glucocorticoid-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
by	NN	O	O
combination	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
with	NN	O	O
AZT	NN	O	O
.	NN	O	O

The	NN	O	O
small	NN	O	O
molecule	NN	O	O
S9a	NN	O	O
was	NN	O	O
derived	NN	O	O
from	NN	O	O
an	NN	O	O
established	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
Canventol	NN	O	O
)	NN	O	O
by	NN	O	O
replacement	NN	O	O
of	NN	O	O
the	NN	O	O
isopropylidine	NN	O	O
group	NN	O	O
with	NN	O	O
a	NN	O	O
phenyl	NN	O	O
ring	NN	O	O
.	NN	O	O

S9a	NN	O	O
at	NN	O	O
10	NN	O	O
to	NN	O	O
100	NN	O	O
nM	NN	O	O
inhibited	NN	O	O
HIV	NN	O	O
production	NN	O	O
as	NN	O	O
potently	NN	O	O
as	NN	O	O
3'-azido-3'-deoxythymidine	NN	O	O
(	NN	O	O
AZT	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
viral	NN	O	B-protein
reverse	NN	O	I-protein
transcriptase	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
S9a	NN	O	O
and	NN	O	O
AZT	NN	O	O
in	NN	O	O
combination	NN	O	O
,	NN	O	O
at	NN	O	O
noncytoxic	NN	O	O
concentrations	NN	O	O
strongly	NN	O	O
inhibited	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
that	NN	O	O
was	NN	O	O
more	NN	O	O
than	NN	O	O
additive	NN	O	O
and	NN	O	O
substantially	NN	O	O
prolonged	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
virus	NN	O	O
both	NN	O	O
in	NN	O	O
acutely	NN	O	B-cell_type
infected	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
SupT	NN	O	B-cell_line
)	NN	O	O
in	NN	O	O
culture	NN	O	O
and	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMCs	NN	O	B-cell_type
)	NN	O	O
infected	NN	O	O
with	NN	O	O
a	NN	O	O
primary	NN	O	O
HIV-1	NN	O	O
isolate	NN	O	O
.	NN	O	O

S9a	NN	O	O
inhibited	NN	O	O
TNF-alpha	NN	O	B-protein
promoter-driven	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
proposed	NN	O	O
that	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
antiviral	NN	O	O
action	NN	O	O
of	NN	O	O
S9a	NN	O	O
was	NN	O	O
on	NN	O	O
the	NN	O	O
host	NN	O	O
cell	NN	O	O
,	NN	O	O
by	NN	O	O
blocking	NN	O	O
TNF-alpha	NN	O	B-protein
transcription	NN	O	O
via	NN	O	O
a	NN	O	O
Tat	NN	O	B-protein
-induced	NN	O	O
tar-independent	NN	O	O
loop	NN	O	O
,	NN	O	O
which	NN	O	O
decreases	NN	O	O
downstream	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

S9a	NN	O	O
was	NN	O	O
superior	NN	O	O
to	NN	O	O
the	NN	O	O
first	NN	O	O
generation	NN	O	O
compound	NN	O	O
Canventol	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
superior	NN	O	O
to	NN	O	O
the	NN	O	O
natural	NN	O	O
compound	NN	O	O
sarcophytol	NN	O	O
A	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
further	NN	O	O
structure-based	NN	O	O
enhancement	NN	O	O
of	NN	O	O
potency	NN	O	O
of	NN	O	O
these	NN	O	O
compounds	NN	O	O
is	NN	O	O
feasible	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
suggests	NN	O	O
a	NN	O	O
therapeutic	NN	O	O
approach	NN	O	O
against	NN	O	O
AIDS	NN	O	O
by	NN	O	O
application	NN	O	O
of	NN	O	O
two	NN	O	O
drugs	NN	O	O
,	NN	O	O
one	NN	O	O
against	NN	O	O
a	NN	O	O
cellular	NN	O	O
and	NN	O	O
the	NN	O	O
other	NN	O	O
a	NN	O	O
viral	NN	O	O
target	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
provide	NN	O	O
an	NN	O	O
approach	NN	O	O
to	NN	O	O
the	NN	O	O
problem	NN	O	O
of	NN	O	O
frequent	NN	O	O
emergence	NN	O	O
of	NN	O	O
resistant	NN	O	O
variants	NN	O	O
to	NN	O	O
combinations	NN	O	O
of	NN	O	O
drugs	NN	O	O
that	NN	O	O
target	NN	O	O
only	NN	O	O
HIV	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Highly	NN	O	O
polarized	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigen	NN	O	I-protein
processing	NN	O	O
and	NN	O	O
presentation	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
intestinal	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
high	NN	O	O
concentration	NN	O	O
of	NN	O	O
foreign	NN	O	B-protein
antigen	NN	O	I-protein
in	NN	O	O
the	NN	O	O
lumen	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
gastrointestinal	NN	O	O
tract	NN	O	O
is	NN	O	O
separated	NN	O	O
from	NN	O	O
the	NN	O	O
underlying	NN	O	O
lymphocytes	NN	O	B-cell_type
by	NN	O	O
a	NN	O	O
single	NN	O	O
cell	NN	O	O
layer	NN	O	O
of	NN	O	O
polarized	NN	O	O
epithelium	NN	O	O
.	NN	O	O

Intestinal	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
express	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
and	NN	O	O
may	NN	O	O
function	NN	O	O
as	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	I-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
within	NN	O	O
the	NN	O	O
intestinal	NN	O	O
mucosa	NN	O	O
.	NN	O	O

Using	NN	O	O
tetanus	NN	O	B-cell_type
toxoid	NN	O	I-cell_type
specific	NN	O	I-cell_type
and	NN	O	I-cell_type
HLA-DR-restricted	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
polarized	NN	O	B-cell_type
intestinal	NN	O	I-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
directed	NN	O	O
to	NN	O	O
express	NN	O	O
HLA-DR	NN	O	B-protein
molecules	NN	O	I-protein
are	NN	O	O
able	NN	O	O
to	NN	O	O
initiate	NN	O	O
class	NN	O	O
II	NN	O	O
processing	NN	O	O
only	NN	O	O
after	NN	O	O
internalization	NN	O	O
of	NN	O	O
antigen	NN	O	O
from	NN	O	O
their	NN	O	O
apical	NN	O	O
surface	NN	O	O
.	NN	O	O

Coexpression	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
CIITA	NN	O	I-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
which	NN	O	O
stimulates	NN	O	O
highly	NN	O	O
efficient	NN	O	O
class	NN	O	O
II	NN	O	O
processing	NN	O	O
without	NN	O	O
the	NN	O	O
characteristic	NN	O	O
decline	NN	O	O
in	NN	O	O
barrier	NN	O	O
function	NN	O	O
seen	NN	O	O
in	NN	O	O
polarized	NN	O	O
monolayers	NN	O	O
treated	NN	O	O
with	NN	O	O
the	NN	O	O
proinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
gamma-IFN	NN	O	I-protein
,	NN	O	O
facilitates	NN	O	O
antigen	NN	O	O
processing	NN	O	O
from	NN	O	O
the	NN	O	O
basolateral	NN	O	O
surface	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
cases	NN	O	O
,	NN	O	O
peptide	NN	O	O
presentation	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
the	NN	O	O
basolateral	NN	O	O
surface	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
a	NN	O	O
highly	NN	O	O
polarized	NN	O	O
functional	NN	O	O
architecture	NN	O	O
for	NN	O	O
antigen	NN	O	O
processing	NN	O	O
and	NN	O	O
presentation	NN	O	O
by	NN	O	O
intestinal	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
functional	NN	O	O
outcome	NN	O	O
of	NN	O	O
antigen	NN	O	O
processing	NN	O	O
by	NN	O	O
the	NN	O	O
intestinal	NN	O	O
epithelium	NN	O	O
is	NN	O	O
both	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
cellular	NN	O	O
surface	NN	O	O
at	NN	O	O
which	NN	O	O
the	NN	O	O
foreign	NN	O	O
antigen	NN	O	O
is	NN	O	O
internalized	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
underlying	NN	O	O
degree	NN	O	O
of	NN	O	O
mucosal	NN	O	O
inflammation	NN	O	O

-DOCSTART-	O

Tobacco	NN	O	O
smoke	NN	O	O
induces	NN	O	O
coordinate	NN	O	O
activation	NN	O	O
of	NN	O	O
HSF	NN	O	B-protein
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
:	NN	O	O
effects	NN	O	O
on	NN	O	O
TNFalpha	NN	O	B-protein
release	NN	O	O
.	NN	O	O

Tobacco	NN	O	O
smoke	NN	O	O
(	NN	O	O
TS	NN	O	O
)	NN	O	O
exposure	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
risk	NN	O	O
factor	NN	O	O
for	NN	O	O
human	NN	O	O
disease	NN	O	O
,	NN	O	O
and	NN	O	O
macrophages	NN	O	B-cell_type
of	NN	O	O
healthy	NN	O	O
smokers	NN	O	O
have	NN	O	O
a	NN	O	O
depressed	NN	O	O
capacity	NN	O	O
to	NN	O	O
release	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
including	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

TS	NN	O	O
induces	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
(	NN	O	I-protein
HS	NN	O	I-protein
)	NN	O	I-protein
/stress	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
HSP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
of	NN	O	O
Hsp70	NN	O	B-protein
.	NN	O	O

We	NN	O	O
determined	NN	O	O
whether	NN	O	O
Hsp70	NN	O	B-protein
induction	NN	O	O
by	NN	O	O
TS	NN	O	O
was	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HS	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
HSF	NN	O	B-protein
.	NN	O	O

HSF	NN	O	B-protein
activation	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
inhibit	NN	O	O
NFkappaB	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
we	NN	O	O
also	NN	O	O
determined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
TS	NN	O	O
on	NN	O	O
NFkappaB	NN	O	B-protein
.	NN	O	O

U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
were	NN	O	O
exposed	NN	O	O
to	NN	O	O
TS	NN	O	O
,	NN	O	O
binding	NN	O	O
activities	NN	O	O
of	NN	O	O
the	NN	O	O
respective	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
were	NN	O	O
analyzed	NN	O	O
,	NN	O	O
and	NN	O	O
Hsp70	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
TNFalpha	NN	O	B-protein
release	NN	O	O
were	NN	O	O
determined	NN	O	O
in	NN	O	O
parallel	NN	O	O
.	NN	O	O

TS	NN	O	O
activated	NN	O	O
HSF	NN	O	B-protein
,	NN	O	O
which	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
Hsp70	NN	O	B-protein
overexpression	NN	O	O
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
TNFalpha	NN	O	B-protein
release	NN	O	O
.	NN	O	O

The	NN	O	O
altered	NN	O	O
cytokine	NN	O	B-protein
profile	NN	O	I-protein
observed	NN	O	O
in	NN	O	O
smokers	NN	O	O
may	NN	O	O
relate	NN	O	O
to	NN	O	O
an	NN	O	O
HSF	NN	O	B-protein
/	NN	O	O
Hsp70	NN	O	B-protein
-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_line
and	NN	O	O
Th2	NN	O	B-cell_line
]	NN	O	O

Upon	NN	O	O
activation	NN	O	O
by	NN	O	O
antigens	NN	O	B-protein
,	NN	O	O
helper	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
differentiate	NN	O	O
into	NN	O	O
one	NN	O	O
of	NN	O	O
several	NN	O	O
subsets	NN	O	O
,	NN	O	O
characterized	NN	O	O
by	NN	O	O
their	NN	O	O
distinct	NN	O	O
cytokine	NN	O	B-protein
-production	NN	O	O
patterns	NN	O	O
.	NN	O	O

Among	NN	O	O
these	NN	O	O
subsets	NN	O	O
,	NN	O	O
Th1	NN	O	B-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
known	NN	O	O
to	NN	O	O
activate	NN	O	O
cellular	NN	O	O
immunity	NN	O	O
resulting	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
,	NN	O	O
whereas	NN	O	O
Th2	NN	O	B-cell_line
cells	NN	O	I-cell_line
induce	NN	O	O
humoral	NN	O	O
and	NN	O	O
allergic	NN	O	O
responses	NN	O	O
and	NN	O	O
suppress	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Th1	NN	O	O
and	NN	O	O
Th2	NN	O	O
effector	NN	O	O
functions	NN	O	O
and	NN	O	O
their	NN	O	O
development	NN	O	O
are	NN	O	O
attributable	NN	O	O
to	NN	O	O
their	NN	O	O
distinct	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
patterns	NN	O	O
.	NN	O	O

Recent	NN	O	O
reports	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
differential	NN	O	O
expression	NN	O	O
of	NN	O	O
cell	NN	O	O
surface	NN	O	O
molecules	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
adhesion	NN	O	B-protein
molecule	NN	O	I-protein
and	NN	O	O
chemokine	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
their	NN	O	O
recruitment	NN	O	O
into	NN	O	O
target	NN	O	O
tissues	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
suggested	NN	O	O
that	NN	O	O
clarification	NN	O	O
of	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
differential	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
Th1/Th2	NN	O	B-cell_line
should	NN	O	O
lead	NN	O	O
to	NN	O	O
rational	NN	O	O
strategies	NN	O	O
for	NN	O	O
manipulating	NN	O	O
pathological	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
helper	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
receptor	NN	O	O
induces	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
biochemical	NN	O	O
events	NN	O	O
.	NN	O	O

Among	NN	O	O
them	NN	O	O
,	NN	O	O
both	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC/Ras-	NN	O	O
and	NN	O	O
CaM/CN-mediated	NN	O	O
pathways	NN	O	O
play	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Closer	NN	O	O
examination	NN	O	O
using	NN	O	O
non-transformed	NN	O	B-cell_line
murine	NN	O	I-cell_line
Th1	NN	O	I-cell_line
and	NN	O	I-cell_line
Th2	NN	O	I-cell_line
clones	NN	O	I-cell_line
suggested	NN	O	O
that	NN	O	O
a	NN	O	O
balance	NN	O	O
between	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
contributes	NN	O	O
to	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
targets	NN	O	O
of	NN	O	O
PGE2	NN	O	O
,	NN	O	O
whose	NN	O	O
effect	NN	O	O
distinguishes	NN	O	O
Th1	NN	O	B-cell_line
from	NN	O	O
Th2	NN	O	O
,	NN	O	O
resides	NN	O	O
in	NN	O	O
the	NN	O	O
downstream	NN	O	O
PKC/Ras	NN	O	B-protein
-mediated	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signalling	NN	O	O
into	NN	O	O
the	NN	O	O
T-cell	NN	O	O
nucleus	NN	O	O
:	NN	O	O
NFAT	NN	O	B-protein
regulation	NN	O	O
.	NN	O	O

The	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
T-cell	NN	O	O
biology	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
T	NN	O	O
cells	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
rapid	NN	O	O
calcineurin	NN	O	B-protein
-dependent	NN	O	O
translocation	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
from	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

This	NN	O	O
translocation	NN	O	O
process	NN	O	O
coupled	NN	O	O
to	NN	O	O
the	NN	O	O
subsequent	NN	O	O
active	NN	O	O
maintenance	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
compartment	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
genes	NN	O	O
encoding	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
membrane	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
modulate	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
cloning	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
has	NN	O	O
facilitated	NN	O	O
rapid	NN	O	O
progress	NN	O	O
in	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
signalling	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
RNA	NN	O	O
display	NN	O	O
identifies	NN	O	O
novel	NN	O	B-DNA
genes	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
decreased	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

To	NN	O	O
characterize	NN	O	O
further	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
vitamin	NN	O	I-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
developed	NN	O	O
stable	NN	O	B-cell_line
transfectant	NN	O	I-cell_line
variants	NN	O	I-cell_line
of	NN	O	O
a	NN	O	O
vitamin	NN	O	B-cell_line
D-responsive	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
U937	NN	O	B-cell_line
)	NN	O	O
which	NN	O	O
express	NN	O	O
either	NN	O	O
decreased	NN	O	O
or	NN	O	O
increased	NN	O	O
numbers	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
have	NN	O	O
analyzed	NN	O	O
changes	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
associated	NN	O	O
with	NN	O	O
this	NN	O	O
variable	NN	O	O
VDR	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Initial	NN	O	O
experiments	NN	O	O
indicated	NN	O	O
that	NN	O	O
a	NN	O	O
50	NN	O	O
%	NN	O	O
decrease	NN	O	O
in	NN	O	O
VDR	NN	O	B-protein
levels	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
2-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
a	NN	O	O
similar	NN	O	O
rise	NN	O	O
in	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Further	NN	O	O
studies	NN	O	O
were	NN	O	O
carried	NN	O	O
out	NN	O	O
using	NN	O	O
differential	NN	O	O
RNA	NN	O	O
display	NN	O	O
(	NN	O	O
DD	NN	O	O
)	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
DD	NN	O	B-protein
products	NN	O	I-protein
revealed	NN	O	O
two	NN	O	O
cDNAs	NN	O	B-DNA
with	NN	O	O
identity	NN	O	O
to	NN	O	O
known	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
:	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
sub-unit	NN	O	I-protein
of	NN	O	I-protein
DNA-protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
DNA-PK	NN	O	B-protein
(	NN	O	I-protein
CS	NN	O	I-protein
)	NN	O	I-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
peroxisomal	NN	O	B-protein
enzyme	NN	O	I-protein
17beta-hydroxysteroid	NN	O	I-protein
dehydrogenase	NN	O	I-protein
type	NN	O	I-protein
IV	NN	O	I-protein
(	NN	O	O
17beta-HSD	NN	O	B-protein
IV	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Northern	NN	O	O
analysis	NN	O	O
confirmed	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
mRNAs	NN	O	B-RNA
was	NN	O	O
reduced	NN	O	O
in	NN	O	O
cells	NN	O	O
with	NN	O	O
decreased	NN	O	O
numbers	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
.	NN	O	O

Down-regulation	NN	O	O
of	NN	O	O
17beta-HSD	NN	O	B-protein
IV	NN	O	I-protein
mRNA	NN	O	O
expression	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
enhanced	NN	O	O
estradiol	NN	O	O
inactivation	NN	O	O
by	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
link	NN	O	O
between	NN	O	O
estrogenic	NN	O	O
pathways	NN	O	O
and	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Further	NN	O	O
Northern	NN	O	O
analyses	NN	O	O
indicated	NN	O	O
that	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
significant	NN	O	O
change	NN	O	O
in	NN	O	O
17beta-HSD	NN	O	B-protein
IV	NN	O	I-protein
or	NN	O	O
DNA-PK	NN	O	B-RNA
(	NN	O	I-RNA
CS	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
following	NN	O	O
treatment	NN	O	O
with	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
,	NN	O	O
although	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
genes	NN	O	O
varied	NN	O	O
with	NN	O	O
changes	NN	O	O
in	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
its	NN	O	O
established	NN	O	O
role	NN	O	O
as	NN	O	O
a	NN	O	O
hormone-dependent	NN	O	B-protein
trans-activator	NN	O	I-protein
,	NN	O	O
VDR	NN	O	B-protein
may	NN	O	O
influence	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
ligand-independent	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

-DOCSTART-	O

Prostaglandin	NN	O	O
E2	NN	O	O
Up-regulates	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
-driven	NN	O	O
gene	NN	O	O
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
NF-kappaB-dependent	NN	O	O
and	NN	O	O
-independent	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

Replication	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type-1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
is	NN	O	O
highly	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
state	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
modulated	NN	O	O
by	NN	O	O
interactions	NN	O	O
between	NN	O	O
viral	NN	O	B-protein
and	NN	O	I-protein
host	NN	O	I-protein
cellular	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Prostaglandin	NN	O	O
E2	NN	O	O
(	NN	O	O
PGE2	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
pleiotropic	NN	O	B-protein
immunomodulatory	NN	O	I-protein
molecule	NN	O	I-protein
,	NN	O	O
is	NN	O	O
observed	NN	O	O
at	NN	O	O
elevated	NN	O	O
levels	NN	O	O
during	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
during	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
other	NN	O	O
pathogenic	NN	O	O
infections	NN	O	O
.	NN	O	O

In	NN	O	O
1G5	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
Jurkat-derived	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
luciferase	NN	O	B-DNA
gene	NN	O	I-DNA
driven	NN	O	O
by	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
PGE2	NN	O	O
markedly	NN	O	O
enhanced	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
-mediated	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

Experiments	NN	O	O
have	NN	O	O
been	NN	O	O
conducted	NN	O	O
to	NN	O	O
identify	NN	O	O
second	NN	O	B-protein
messengers	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
this	NN	O	O
PGE2-dependent	NN	O	O
up-regulating	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
regulatory	NN	O	O
element	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
present	NN	O	O
evidence	NN	O	O
indicating	NN	O	O
that	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
induced	NN	O	O
by	NN	O	O
PGE2	NN	O	O
necessitate	NN	O	O
the	NN	O	O
participation	NN	O	O
of	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
,	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
.	NN	O	O

Experiments	NN	O	O
conducted	NN	O	O
with	NN	O	O
different	NN	O	O
HIV-1	NN	O	B-DNA
LTR-based	NN	O	I-DNA
vectors	NN	O	I-DNA
suggested	NN	O	O
that	NN	O	O
PGE2-mediated	NN	O	O
activation	NN	O	O
effect	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
was	NN	O	O
transduced	NN	O	O
via	NN	O	O
both	NN	O	O
NF-kappaB-dependent	NN	O	O
and	NN	O	O
-independent	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
the	NN	O	O
PGE2-dependent	NN	O	O
activating	NN	O	O
effect	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
was	NN	O	O
further	NN	O	O
confirmed	NN	O	O
using	NN	O	O
a	NN	O	O
kappaB-regulated	NN	O	B-DNA
luciferase	NN	O	I-DNA
encoding	NN	O	I-DNA
vector	NN	O	I-DNA
and	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

Results	NN	O	O
from	NN	O	O
Northern	NN	O	O
blot	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometric	NN	O	O
analyses	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
selective	NN	O	O
antagonist	NN	O	O
indicated	NN	O	O
that	NN	O	O
PGE2	NN	O	O
modulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
-driven	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
studied	NN	O	O
T	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
transduced	NN	O	O
via	NN	O	O
the	NN	O	O
EP4	NN	O	B-protein
receptor	NN	O	I-protein
subtype	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
secretion	NN	O	O
of	NN	O	O
PGE2	NN	O	O
by	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
infection	NN	O	O
or	NN	O	O
inflammatory	NN	O	O
activators	NN	O	O
could	NN	O	O
induce	NN	O	O
signaling	NN	O	O
events	NN	O	O
resulting	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
proviral	NN	O	O
DNA	NN	O	O
present	NN	O	O
into	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
latently	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
distinct	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
neutrophils	NN	O	B-cell_type
by	NN	O	O
bacterial	NN	O	O
LPS	NN	O	O
,	NN	O	O
interferon-gamma	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GM-CSF	NN	O	B-protein
and	NN	O	O
the	NN	O	O
necessity	NN	O	O
to	NN	O	O
overcome	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
proteases	NN	O	I-protein
.	NN	O	O

Human	NN	O	O
neutrophils	NN	O	B-cell_type
can	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
actively	NN	O	O
transcribe	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
early-response	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
particular	NN	O	O
those	NN	O	O
encoding	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
chemokines	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
high-affinity	NN	O	O
surface	NN	O	O
receptor	NN	O	O
for	NN	O	O
IgG	NN	O	B-protein
,	NN	O	O
FcgammaRI	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
date	NN	O	O
about	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
several	NN	O	O
indications	NN	O	O
point	NN	O	O
to	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
distinct	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
STAT	NN	O	B-protein
families	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
become	NN	O	O
activated	NN	O	O
under	NN	O	O
stimulatory	NN	O	O
conditions	NN	O	O
which	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
induce	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
conventional	NN	O	O
procedures	NN	O	O
employed	NN	O	O
to	NN	O	O
prepare	NN	O	O
cellular	NN	O	O
extracts	NN	O	O
cause	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
proteolytic	NN	O	O
activities	NN	O	O
that	NN	O	O
are	NN	O	O
normally	NN	O	O
stored	NN	O	O
in	NN	O	O
intracellular	NN	O	O
granules	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
various	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
and	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

To	NN	O	O
circumvent	NN	O	O
this	NN	O	O
problem	NN	O	O
,	NN	O	O
we	NN	O	O
developed	NN	O	O
an	NN	O	O
alternative	NN	O	O
procedure	NN	O	O
which	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
LPS	NN	O	O
and	NN	O	O
TNFalpha	NN	O	B-protein
induce	NN	O	O
a	NN	O	O
NF-kappaB	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
which	NN	O	O
essentially	NN	O	O
consists	NN	O	O
of	NN	O	O
p50/RelA	NN	O	B-protein
dimers	NN	O	I-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
IFNgamma	NN	O	B-protein
promotes	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
STAT1	NN	O	B-protein
homodimers	NN	O	I-protein
to	NN	O	O
the	NN	O	O
IFNgamma	NN	O	B-DNA
response	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
FcgammaRI	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
neutrophil	NN	O	O
stimulation	NN	O	O
with	NN	O	O
GM-CSF	NN	O	B-protein
results	NN	O	O
in	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
STAT5	NN	O	B-protein
-containing	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
the	NN	O	O
current	NN	O	O
findings	NN	O	O
open	NN	O	O
new	NN	O	O
perspectives	NN	O	O
about	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
regulate	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
neutrophils	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
procedure	NN	O	O
described	NN	O	O
herein	NN	O	O
could	NN	O	O
prove	NN	O	O
useful	NN	O	O
in	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
that	NN	O	O
express	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
proteases	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Low	NN	O	O
CD3+CD28-induced	NN	O	O
interleukin-2	NN	O	B-protein
production	NN	O	O
correlates	NN	O	O
with	NN	O	O
decreased	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediate	NN	O	O
formation	NN	O	O
in	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
capacity	NN	O	O
of	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
secrete	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
variable	NN	O	O
.	NN	O	O

We	NN	O	O
analysed	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
in	NN	O	O
purified	NN	O	O
neonatal	NN	O	B-cell_type
and	NN	O	I-cell_type
adult	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
polyclonal	NN	O	O
activator	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
+	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
(	NN	O	O
PDBu	NN	O	O
+	NN	O	O
iono	NN	O	O
)	NN	O	O
or	NN	O	O
receptor-mediated	NN	O	O
anti-CD3	NN	O	B-protein
/anti-CD3+	NN	O	O
anti-CD28	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

PDBu	NN	O	O
+	NN	O	O
iono	NN	O	O
induced	NN	O	O
equally	NN	O	O
high	NN	O	O
IL-2	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
both	NN	O	O
groups	NN	O	O
and	NN	O	O
,	NN	O	O
when	NN	O	O
stimulated	NN	O	O
with	NN	O	O
plate-bound	NN	O	O
anti-CD3	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
mAb	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
neonatal	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
undetectable	NN	O	O
and	NN	O	O
adult	NN	O	B-cell_type
cells	NN	O	I-cell_type
produced	NN	O	O
low	NN	O	O
amounts	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
(	NN	O	O
mean	NN	O	O
331	NN	O	O
+/-	NN	O	O
86	NN	O	O
pg/ml	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
to	NN	O	O
anti-CD3-stimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
markedly	NN	O	O
increased	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
but	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
remained	NN	O	O
clearly	NN	O	O
lower	NN	O	O
than	NN	O	O
those	NN	O	O
of	NN	O	O
adult	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
respective	NN	O	O
mean	NN	O	O
values	NN	O	O
:	NN	O	O
385	NN	O	O
+/-	NN	O	O
109	NN	O	O
pg/ml	NN	O	O
and	NN	O	O
4494	NN	O	O
+/-	NN	O	O
1199	NN	O	O
pg/ml	NN	O	O
)	NN	O	O
.	NN	O	O

As	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
critical	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
we	NN	O	O
next	NN	O	O
analysed	NN	O	O
its	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
in	NN	O	O
neonatal	NN	O	B-cell_type
and	NN	O	I-cell_type
adult	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
and	NN	O	O
,	NN	O	O
because	NN	O	O
induction	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
(	NN	O	O
ROI	NN	O	O
)	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
we	NN	O	O
also	NN	O	O
analysed	NN	O	O
levels	NN	O	O
of	NN	O	O
intracellular	NN	O	O
ROI	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
using	NN	O	O
the	NN	O	O
ROI-reactive	NN	O	O
fluorochrome	NN	O	O
DCFH-DA	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

In	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
ROI	NN	O	O
formation	NN	O	O
after	NN	O	O
anti-CD3	NN	O	B-protein
stimulation	NN	O	O
were	NN	O	O
low	NN	O	O
compared	NN	O	O
with	NN	O	O
adult	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
,	NN	O	O
although	NN	O	O
addition	NN	O	O
of	NN	O	O
anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
increased	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
ROI	NN	O	O
formation	NN	O	O
,	NN	O	O
levels	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
adults	NN	O	O
were	NN	O	O
not	NN	O	O
achieved	NN	O	O
.	NN	O	O

After	NN	O	O
PDBu	NN	O	O
+	NN	O	O
iono	NN	O	O
stimulation	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
showed	NN	O	O
similar	NN	O	O
ROI	NN	O	O
formation	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
reduced	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
by	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
specific	NN	O	O
for	NN	O	O
anti-CD3	NN	O	B-protein
and	NN	O	O
anti-CD3	NN	O	B-protein
+	NN	O	O
anti-CD28	NN	O	B-protein
-mediated	NN	O	O
stimulation	NN	O	O
and	NN	O	O
that	NN	O	O
these	NN	O	O
activators	NN	O	O
can	NN	O	O
not	NN	O	O
effectively	NN	O	O
activate	NN	O	O
the	NN	O	O
ROI	NN	O	O
-NF-kappa	NN	O	B-protein
B	NN	O	I-protein
signalling	NN	O	O
pathway	NN	O	O
in	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

CD30	NN	O	B-protein
is	NN	O	O
a	NN	O	O
CD40-inducible	NN	O	B-protein
molecule	NN	O	I-protein
that	NN	O	O
negatively	NN	O	O
regulates	NN	O	O
CD40-mediated	NN	O	B-protein
immunoglobulin	NN	O	I-protein
class	NN	O	O
switching	NN	O	O
in	NN	O	O
non-antigen-selected	NN	O	B-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
used	NN	O	O
our	NN	O	O
monoclonal	NN	O	O
model	NN	O	O
of	NN	O	O
germinal	NN	O	O
center	NN	O	O
maturation	NN	O	O
,	NN	O	O
CL-01	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
CD30	NN	O	B-protein
in	NN	O	O
human	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

CL-01	NN	O	B-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
IgM	NN	O	B-protein
+	NN	O	O
IgD	NN	O	B-protein
+	NN	O	O
CD30	NN	O	B-protein
+	NN	O	O
and	NN	O	O
switch	NN	O	O
to	NN	O	O
IgG	NN	O	B-protein
,	NN	O	O
IgA	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IgE	NN	O	B-protein
when	NN	O	O
exposed	NN	O	O
to	NN	O	O
CD40L	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

Switching	NN	O	O
is	NN	O	O
hampered	NN	O	O
by	NN	O	O
CD30	NN	O	B-protein
coengagement	NN	O	O
,	NN	O	O
possibly	NN	O	O
through	NN	O	O
interference	NN	O	O
with	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
downstream	NN	O	B-DNA
C	NN	O	I-DNA
(	NN	O	I-DNA
H	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
physiological	NN	O	O
relevance	NN	O	O
of	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
is	NN	O	O
emphasized	NN	O	O
by	NN	O	O
similar	NN	O	O
CD30	NN	O	B-protein
-mediated	NN	O	O
effects	NN	O	O
in	NN	O	O
naive	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
CD30	NN	O	B-protein
by	NN	O	O
these	NN	O	O
cells	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
CD40L	NN	O	B-protein
but	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
B	NN	O	O
cell	NN	O	O
receptor	NN	O	O
coengagement	NN	O	O
and/or	NN	O	O
exposure	NN	O	O
to	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
IL-12	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
CD30	NN	O	B-protein
critically	NN	O	O
regulates	NN	O	O
the	NN	O	O
CD40	NN	O	B-protein
-mediated	NN	O	O
differentiation	NN	O	O
of	NN	O	O
non-antigen-selected	NN	O	B-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Calcineurin	NN	O	B-protein
and	NN	O	O
the	NN	O	O
biological	NN	O	O
effect	NN	O	O
of	NN	O	O
cyclosporine	NN	O	O
and	NN	O	O
tacrolimus	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
effect	NN	O	O
of	NN	O	O
CyA	NN	O	O
and	NN	O	O
FK	NN	O	O
506	NN	O	O
can	NN	O	O
be	NN	O	O
monitored	NN	O	O
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
humans	NN	O	O
.	NN	O	O

The	NN	O	O
picture	NN	O	O
emerging	NN	O	O
is	NN	O	O
of	NN	O	O
a	NN	O	O
close	NN	O	O
relationship	NN	O	O
between	NN	O	O
drug	NN	O	O
concentrations	NN	O	O
and	NN	O	O
CN	NN	O	O
inhibition	NN	O	O
.	NN	O	O

But	NN	O	O
many	NN	O	O
puzzles	NN	O	O
of	NN	O	O
the	NN	O	O
drugs	NN	O	O
remain	NN	O	O
.	NN	O	O

What	NN	O	O
is	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
CyA	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
(	NN	O	O
TGF-beta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
particularly	NN	O	O
in	NN	O	O
relationship	NN	O	O
to	NN	O	O
nephrotoxicity	NN	O	O
and	NN	O	O
fibrogenesis	NN	O	O
?	NN	O	O

How	NN	O	O
important	NN	O	O
are	NN	O	O
the	NN	O	O
anti-inflammatory	NN	O	O
(	NN	O	O
non	NN	O	O
T	NN	O	O
)	NN	O	O
effects	NN	O	O
of	NN	O	O
CyA	NN	O	O
,	NN	O	O
and	NN	O	O
which	NN	O	O
cells	NN	O	O
do	NN	O	O
they	NN	O	O
operate	NN	O	O
in	NN	O	O
?	NN	O	O

Are	NN	O	O
there	NN	O	O
effects	NN	O	O
of	NN	O	O
CyA	NN	O	O
and	NN	O	O
FK	NN	O	O
506	NN	O	O
all	NN	O	O
attributable	NN	O	O
to	NN	O	O
CN	NN	O	O
inhibition	NN	O	O
,	NN	O	O
and	NN	O	O
how	NN	O	O
much	NN	O	O
of	NN	O	O
them	NN	O	O
are	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
NFATC	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
?	NN	O	O

Finally	NN	O	O
,	NN	O	O
it	NN	O	O
would	NN	O	O
be	NN	O	O
useful	NN	O	O
to	NN	O	O
know	NN	O	O
what	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
CyA	NN	O	O
are	NN	O	O
on	NN	O	O
tolerance	NN	O	O
and	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
events	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stimulation	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
activates	NN	O	O
expression	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
and	NN	O	I-protein
differentiation	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

During	NN	O	O
B	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
proliferation	NN	O	O
many	NN	O	O
genes	NN	O	O
are	NN	O	O
induced	NN	O	O
or	NN	O	O
repressed	NN	O	O
while	NN	O	O
certain	NN	O	O
genes	NN	O	O
are	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
processes	NN	O	O
related	NN	O	O
to	NN	O	O
B	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
the	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
stimulated	NN	O	O
and	NN	O	O
nonstimulated	NN	O	B-cell_line
Ramos	NN	O	I-cell_line
and	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
studied	NN	O	O
using	NN	O	O
cDNA	NN	O	O
microarray	NN	O	O
technology	NN	O	O
.	NN	O	O

Simultaneous	NN	O	O
analysis	NN	O	O
of	NN	O	O
close	NN	O	O
to	NN	O	O
600	NN	O	B-DNA
genes	NN	O	I-DNA
indicated	NN	O	O
highest	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
certain	NN	O	O
transcription	NN	O	B-protein
,	NN	O	I-protein
differentiation	NN	O	I-protein
and	NN	O	I-protein
proliferation	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Many	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
have	NN	O	O
not	NN	O	O
previously	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
funcion	NN	O	O
in	NN	O	O
the	NN	O	O
stimulated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Also	NN	O	O
genes	NN	O	O
encoding	NN	O	O
proteins	NN	O	O
involved	NN	O	O
in	NN	O	O
DNA	NN	O	O
replication	NN	O	O
,	NN	O	O
binding	NN	O	O
,	NN	O	O
transcription	NN	O	O
and	NN	O	O
translation	NN	O	O
were	NN	O	O
induced	NN	O	O
.	NN	O	O

Large	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
activated	NN	O	B-DNA
genes	NN	O	I-DNA
were	NN	O	O
under	NN	O	O
very	NN	O	O
stringent	NN	O	O
regulation	NN	O	O
being	NN	O	O
expressed	NN	O	O
only	NN	O	O
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
and	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
expressed	NN	O	B-DNA
genes	NN	O	I-DNA
during	NN	O	O
lymphocyte	NN	O	O
differentiation	NN	O	O
and	NN	O	O
in	NN	O	O
disorders	NN	O	O
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin	NN	O	B-protein
2	NN	O	I-protein
and	NN	O	I-protein
15	NN	O	I-protein
activate	NN	O	O
Stat3alpha	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
Stat3	NN	O	B-protein
)	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
exist	NN	O	O
in	NN	O	O
two	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
isoforms	NN	O	O
,	NN	O	O
a	NN	O	O
short	NN	O	O
form	NN	O	O
,	NN	O	O
Stat3beta	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
longer	NN	O	O
form	NN	O	O
,	NN	O	O
Stat3alpha	NN	O	B-protein
,	NN	O	O
displaying	NN	O	O
differences	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
unknown	NN	O	O
which	NN	O	O
Stat3	NN	O	B-protein
isoform	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
is	NN	O	O
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
and	NN	O	O
IL-15	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
in	NN	O	O
previously	NN	O	O
activated	NN	O	O
CD4	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
examined	NN	O	O
using	NN	O	O
Stat3	NN	O	B-protein
antibodies	NN	O	O
directed	NN	O	O
against	NN	O	O
different	NN	O	O
regions	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
.	NN	O	O

As	NN	O	O
determined	NN	O	O
by	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
binding	NN	O	O
to	NN	O	O
an	NN	O	O
hSIE-oligonucleotide	NN	O	O
probe	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-15	NN	O	B-protein
activated	NN	O	O
the	NN	O	O
slowly	NN	O	O
migrating	NN	O	O
isoform	NN	O	O
,	NN	O	O
Stat3alpha	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
minimal	NN	O	O
or	NN	O	O
no	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat3beta	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-15	NN	O	B-protein
predominantly	NN	O	O
activate	NN	O	O
Stat3alpha	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
CD4	NN	O	I-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
way	NN	O	O
,	NN	O	O
diversity	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
proteins	NN	O	I-protein
may	NN	O	O
provide	NN	O	O
additional	NN	O	O
means	NN	O	O
of	NN	O	O
regulating	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
lysosomal	NN	O	B-protein
acid	NN	O	I-protein
lipase	NN	O	I-protein
in	NN	O	O
differentiating	NN	O	O
monocytes	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
Sp1	NN	O	B-protein
and	NN	O	O
AP-2	NN	O	B-protein
.	NN	O	O

Human	NN	O	O
lysosomal	NN	O	B-protein
acid	NN	O	I-protein
lipase	NN	O	I-protein
(	NN	O	O
LAL	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
hydrolase	NN	O	B-protein
required	NN	O	O
for	NN	O	O
the	NN	O	O
cleavage	NN	O	O
of	NN	O	O
cholesteryl	NN	O	O
esters	NN	O	O
and	NN	O	O
triglycerides	NN	O	O
derived	NN	O	O
from	NN	O	O
plasma	NN	O	O
lipoproteins	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
shown	NN	O	O
here	NN	O	O
that	NN	O	O
during	NN	O	O
monocyte	NN	O	O
to	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
LAL-mRNA	NN	O	B-RNA
is	NN	O	O
induced	NN	O	O
.	NN	O	O

This	NN	O	O
induction	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
activity	NN	O	O
and	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

The	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
increase	NN	O	O
in	NN	O	O
LAL	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
further	NN	O	O
investigated	NN	O	O
in	NN	O	O
the	NN	O	O
THP-1	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
with	NN	O	O
respect	NN	O	O
to	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	B-cell_line
differentiates	NN	O	O
into	NN	O	O
macrophage-like	NN	O	B-cell_type
cells	NN	O	I-cell_type
when	NN	O	O
treated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
necessary	NN	O	O
for	NN	O	O
both	NN	O	O
basal	NN	O	O
and	NN	O	O
phorbol	NN	O	O
12-myristate-13	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
-enhanced	NN	O	O
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
we	NN	O	O
performed	NN	O	O
deletion	NN	O	O
analysis	NN	O	O
and	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
assays	NN	O	O
.	NN	O	O

A	NN	O	O
PMA	NN	O	B-DNA
responsive	NN	O	I-DNA
element	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
identified	NN	O	O
between	NN	O	O
-182	NN	O	B-DNA
bp	NN	O	I-DNA
and	NN	O	O
-107	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
transcription	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
AP-2	NN	O	B-protein
to	NN	O	O
the	NN	O	O
LAL	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
increased	NN	O	O
by	NN	O	O
PMA	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Co-transfections	NN	O	O
with	NN	O	O
expression	NN	O	B-DNA
plasmids	NN	O	I-DNA
for	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
AP-2	NN	O	B-protein
further	NN	O	O
emphasized	NN	O	O
the	NN	O	O
important	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
both	NN	O	O
basal	NN	O	O
and	NN	O	O
PMA-enhanced	NN	O	O
LAL	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
differentiation	NN	O	O
dependent	NN	O	O
increase	NN	O	O
of	NN	O	O
lysosomal	NN	O	B-protein
acid	NN	O	I-protein
lipase	NN	O	I-protein
(	NN	O	O
LAL	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
concerted	NN	O	O
action	NN	O	O
of	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
AP-2	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Relationship	NN	O	O
between	NN	O	O
IkappaBalpha	NN	O	B-protein
constitutive	NN	O	O
expression	NN	O	O
,	NN	O	O
TNFalpha	NN	O	B-protein
synthesis	NN	O	O
,	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
EBV-infected	NN	O	B-cell_type
lymphoblastoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
EBV	NN	O	O
transformation	NN	O	O
we	NN	O	O
have	NN	O	O
established	NN	O	O
stably	NN	O	O
transfected	NN	O	O
IkappaBalpha	NN	O	B-protein
into	NN	O	O
lymphoblastoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Two	NN	O	O
clones	NN	O	O
were	NN	O	O
obtained	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
transgenic	NN	O	O
IkappaBalpha	NN	O	B-protein
.	NN	O	O

Protein	NN	O	O
latency	NN	O	O
expression	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
immunocytochemistry	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
surface	NN	O	O
markers	NN	O	O
,	NN	O	O
intracytoplasmic	NN	O	O
content	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
cell	NN	O	O
cycle	NN	O	O
analysis	NN	O	O
after	NN	O	O
BrdU	NN	O	B-protein
incorporation	NN	O	O
and	NN	O	O
DNA	NN	O	O
staining	NN	O	O
with	NN	O	O
propidium	NN	O	O
iodide	NN	O	O
were	NN	O	O
studied	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

Percentage	NN	O	O
of	NN	O	O
apoptotic	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
determined	NN	O	O
by	NN	O	O
in-situ	NN	O	O
labelling	NN	O	O
of	NN	O	O
DNA	NN	O	O
strand	NN	O	O
breaks	NN	O	O
.	NN	O	O

No	NN	O	O
significative	NN	O	O
changes	NN	O	O
in	NN	O	O
EBV	NN	O	O
latency	NN	O	O
nor	NN	O	O
in	NN	O	O
cell	NN	O	O
surface	NN	O	O
marker	NN	O	O
expression	NN	O	O
was	NN	O	O
found	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
intracytoplasmic	NN	O	O
TNFalpha	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
strongly	NN	O	O
reduced	NN	O	O
in	NN	O	O
transfected	NN	O	B-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
30	NN	O	O
%	NN	O	O
of	NN	O	O
IkappaBalpha	NN	O	B-cell_line
transfected	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
apoptotic	NN	O	O
after	NN	O	O
8	NN	O	O
h	NN	O	O
of	NN	O	O
TNFalpha	NN	O	B-protein
treatment	NN	O	O
.	NN	O	O

This	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
strong	NN	O	O
reduction	NN	O	O
of	NN	O	O
BrdU	NN	O	O
incorporation	NN	O	O
after	NN	O	O
24	NN	O	O
h	NN	O	O
of	NN	O	O
TNFalpha	NN	O	B-protein
treatment	NN	O	O
.	NN	O	O

No	NN	O	O
effect	NN	O	O
was	NN	O	O
seen	NN	O	O
with	NN	O	O
non	NN	O	O
transfected	NN	O	B-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
with	NN	O	O
cells	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
control	NN	O	O
plasmid	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
TNFalpha	NN	O	B-DNA
gene	NN	O	I-DNA
could	NN	O	O
be	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
targets	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
EBV	NN	O	O
infected	NN	O	O
cells	NN	O	O
and	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-cell_type
protects	NN	O	I-cell_type
EBV-infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
TNFalpha	NN	O	B-protein
,	NN	O	O
which	NN	O	O
may	NN	O	O
favour	NN	O	O
the	NN	O	O
proliferative	NN	O	O
effect	NN	O	O
of	NN	O	O
this	NN	O	O
cytokine	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Seminoma	NN	O	O
in	NN	O	O
a	NN	O	O
postmenopausal	NN	O	O
woman	NN	O	O
with	NN	O	O
a	NN	O	O
Y	NN	O	B-DNA
;	NN	O	I-DNA
15	NN	O	I-DNA
translocation	NN	O	I-DNA
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
a	NN	O	O
t	NN	O	O
(	NN	O	O
Y	NN	O	O
;	NN	O	O
15	NN	O	O
)	NN	O	O
/45	NN	O	O
,	NN	O	O
X	NN	O	O
Turner	NN	O	O
mosaic	NN	O	O
pattern	NN	O	O
in	NN	O	O
skin	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
report	NN	O	O
an	NN	O	O
unusual	NN	O	O
case	NN	O	O
of	NN	O	O
a	NN	O	O
55	NN	O	O
year	NN	O	O
old	NN	O	O
Japanese	NN	O	O
woman	NN	O	O
with	NN	O	O
a	NN	O	O
seminoma	NN	O	O
but	NN	O	O
relatively	NN	O	O
normal	NN	O	O
menses	NN	O	O
.	NN	O	O

The	NN	O	O
patient	NN	O	O
was	NN	O	O
a	NN	O	O
phenotypic	NN	O	O
female	NN	O	O
with	NN	O	O
late	NN	O	O
onset	NN	O	O
menarche	NN	O	O
(	NN	O	O
18	NN	O	O
years	NN	O	O
of	NN	O	O
age	NN	O	O
)	NN	O	O
,	NN	O	O
who	NN	O	O
was	NN	O	O
amenorrhoeic	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
year	NN	O	O
,	NN	O	O
followed	NN	O	O
by	NN	O	O
menses	NN	O	O
of	NN	O	O
one	NN	O	O
to	NN	O	O
three	NN	O	O
days	NN	O	O
'	NN	O	O
slight	NN	O	O
flow	NN	O	O
with	NN	O	O
dysmenorrhoea	NN	O	O
,	NN	O	O
but	NN	O	O
an	NN	O	O
otherwise	NN	O	O
normal	NN	O	O
menstrual	NN	O	O
history	NN	O	O
.	NN	O	O

A	NN	O	O
typical	NN	O	O
seminoma	NN	O	O
was	NN	O	O
removed	NN	O	O
from	NN	O	O
the	NN	O	O
left	NN	O	O
adnexal	NN	O	O
region	NN	O	O
and	NN	O	O
an	NN	O	O
immature	NN	O	O
testis	NN	O	O
was	NN	O	O
identified	NN	O	O
separately	NN	O	O
as	NN	O	O
an	NN	O	O
associated	NN	O	O
right	NN	O	O
adnexal	NN	O	O
mass	NN	O	O
.	NN	O	O

Repeated	NN	O	O
karyotypic	NN	O	O
studies	NN	O	O
on	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
cultures	NN	O	I-cell_type
showed	NN	O	O
only	NN	O	O
46	NN	O	O
,	NN	O	O
X	NN	O	O
,	NN	O	O
-Y	NN	O	O
,	NN	O	O
t	NN	O	O
(	NN	O	O
Y	NN	O	O
;	NN	O	O
15	NN	O	O
)	NN	O	O
(	NN	O	O
q12	NN	O	O
;	NN	O	O
p13	NN	O	O
)	NN	O	O
.	NN	O	O

Cytogenetic	NN	O	O
examination	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
younger	NN	O	O
brother	NN	O	O
,	NN	O	O
who	NN	O	O
had	NN	O	O
fathered	NN	O	O
three	NN	O	O
healthy	NN	O	O
children	NN	O	O
,	NN	O	O
showed	NN	O	O
an	NN	O	O
identical	NN	O	O
karyotype	NN	O	O
.	NN	O	O

Mosaicism	NN	O	O
of	NN	O	O
46	NN	O	B-cell_line
,	NN	O	I-cell_line
X	NN	O	I-cell_line
,	NN	O	I-cell_line
-Y	NN	O	I-cell_line
,	NN	O	I-cell_line
t	NN	O	I-cell_line
(	NN	O	I-cell_line
Y	NN	O	I-cell_line
;	NN	O	I-cell_line
15	NN	O	I-cell_line
)	NN	O	I-cell_line
(	NN	O	I-cell_line
q12	NN	O	I-cell_line
;	NN	O	I-cell_line
p13	NN	O	I-cell_line
)	NN	O	I-cell_line
/45	NN	O	I-cell_line
,	NN	O	I-cell_line
X	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
was	NN	O	O
found	NN	O	O
in	NN	O	O
skin	NN	O	O
samples	NN	O	O
from	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
elbow	NN	O	O
and	NN	O	O
genital	NN	O	O
regions	NN	O	O
,	NN	O	O
although	NN	O	O
there	NN	O	O
were	NN	O	O
no	NN	O	O
clinical	NN	O	O
stigmata	NN	O	O
of	NN	O	O
Turner	NN	O	O
syndrome	NN	O	O
.	NN	O	O

An	NN	O	O
androgen	NN	O	O
receptor	NN	O	O
binding	NN	O	O
assay	NN	O	O
of	NN	O	O
cultured	NN	O	B-cell_line
genital	NN	O	I-cell_line
skin	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
was	NN	O	O
negative	NN	O	O
.	NN	O	O

Molecular	NN	O	O
analysis	NN	O	O
using	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridisation	NN	O	O
,	NN	O	O
PCR	NN	O	O
,	NN	O	O
and	NN	O	O
direct	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
showed	NN	O	O
that	NN	O	O
neither	NN	O	O
the	NN	O	O
patient	NN	O	O
nor	NN	O	O
her	NN	O	O
brother	NN	O	O
had	NN	O	O
a	NN	O	O
detectable	NN	O	O
deletion	NN	O	O
or	NN	O	O
other	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
Y	NN	O	O
chromosome	NN	O	O
sequences	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
SRY	NN	O	B-DNA
(	NN	O	I-DNA
sex	NN	O	I-DNA
determining	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	I-DNA
the	NN	O	I-DNA
Y	NN	O	I-DNA
chromosome	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
Turner	NN	O	O
mosaicism	NN	O	O
of	NN	O	O
the	NN	O	O
45	NN	O	B-cell_line
,	NN	O	I-cell_line
X	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
may	NN	O	O
have	NN	O	O
contributed	NN	O	O
to	NN	O	O
this	NN	O	O
atypical	NN	O	O
presentation	NN	O	O
in	NN	O	O
an	NN	O	O
XY	NN	O	O
female	NN	O	O
,	NN	O	O
although	NN	O	O
we	NN	O	O
can	NN	O	O
not	NN	O	O
exclude	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
other	NN	O	O
genes	NN	O	O
related	NN	O	O
to	NN	O	O
sex	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Potent	NN	O	O
inhibition	NN	O	O
of	NN	O	O
HIV	NN	O	O
type	NN	O	O
1	NN	O	O
replication	NN	O	O
by	NN	O	O
an	NN	O	O
antiinflammatory	NN	O	O
alkaloid	NN	O	O
,	NN	O	O
cepharanthine	NN	O	O
,	NN	O	O
in	NN	O	O
chronically	NN	O	B-cell_type
infected	NN	O	I-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cepharanthine	NN	O	O
is	NN	O	O
a	NN	O	O
biscoclaurine	NN	O	O
alkaloid	NN	O	O
isolated	NN	O	O
from	NN	O	O
Stephania	NN	O	O
cepharantha	NN	O	O
Hayata	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
have	NN	O	O
antiinflammatory	NN	O	O
,	NN	O	O
antiallergic	NN	O	O
,	NN	O	O
and	NN	O	O
immunomodulatory	NN	O	O
activities	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

As	NN	O	O
several	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
and	NN	O	O
oxidative	NN	O	O
stresses	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
cepharanthine	NN	O	O
on	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor	NN	O	O
alpha	NN	O	O
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
-	NN	O	O
and	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
-induced	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
chronically	NN	O	B-cell_line
infected	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Two	NN	O	O
chronically	NN	O	B-cell_line
HIV-1-infected	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
U1	NN	O	B-cell_line
(	NN	O	O
monocytic	NN	O	O
)	NN	O	O
and	NN	O	O
ACH-2	NN	O	B-cell_line
(	NN	O	O
T	NN	O	O
lymphocytic	NN	O	O
)	NN	O	O
,	NN	O	O
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
TNF-alpha	NN	O	B-protein
or	NN	O	O
PMA	NN	O	O
and	NN	O	O
cultured	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
various	NN	O	O
concentrations	NN	O	O
of	NN	O	O
the	NN	O	O
compound	NN	O	O
.	NN	O	O

HIV-1	NN	O	O
replication	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
p24	NN	O	B-protein
antigen	NN	O	I-protein
level	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
cepharanthine	NN	O	O
on	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
-driven	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
activation	NN	O	O
were	NN	O	O
also	NN	O	O
examined	NN	O	O
.	NN	O	O

Cepharanthine	NN	O	O
dose	NN	O	O
dependently	NN	O	O
inhibited	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
TNF-alpha-	NN	O	B-cell_line
and	NN	O	I-cell_line
PMA-stimulated	NN	O	I-cell_line
U1	NN	O	I-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
ACH-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Its	NN	O	O
50	NN	O	O
%	NN	O	O
effective	NN	O	O
and	NN	O	O
cytotoxic	NN	O	O
concentrations	NN	O	O
were	NN	O	O
0.016	NN	O	O
and	NN	O	O
2.2	NN	O	O
microg/ml	NN	O	O
in	NN	O	O
PMA-stimulated	NN	O	B-cell_line
U1	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Cepharanthine	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
suppress	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
-driven	NN	O	O
gene	NN	O	O
expression	NN	O	O
through	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
cepharanthine	NN	O	O
is	NN	O	O
a	NN	O	O
highly	NN	O	O
potent	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
a	NN	O	O
chronically	NN	O	B-cell_line
infected	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Since	NN	O	O
biscoclaurine	NN	O	O
alkaloids	NN	O	O
,	NN	O	O
containing	NN	O	O
cepharanthine	NN	O	O
as	NN	O	O
a	NN	O	O
major	NN	O	O
component	NN	O	O
,	NN	O	O
are	NN	O	O
widely	NN	O	O
used	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
various	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
in	NN	O	O
Japan	NN	O	O
,	NN	O	O
cepharanthine	NN	O	O
should	NN	O	O
be	NN	O	O
further	NN	O	O
pursued	NN	O	O
for	NN	O	O
its	NN	O	O
chemotherapeutic	NN	O	O
potential	NN	O	O
in	NN	O	O
HIV-1-infected	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappaB	NN	O	B-protein
only	NN	O	O
partially	NN	O	O
mediates	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
latent	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
activation	NN	O	O
of	NN	O	O
B	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
LMP1	NN	O	B-protein
)	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
EBV-induced	NN	O	O
immortalization	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
causes	NN	O	O
tumorigenic	NN	O	O
transformation	NN	O	O
of	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

LMP1	NN	O	B-protein
expression	NN	O	O
induces	NN	O	O
phenotypic	NN	O	O
changes	NN	O	O
resembling	NN	O	O
B	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
cell	NN	O	O
size	NN	O	O
increase	NN	O	O
and	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
activation	NN	O	I-protein
markers	NN	O	I-protein
.	NN	O	O

LMP1	NN	O	B-protein
contains	NN	O	O
two	NN	O	O
domains	NN	O	O
that	NN	O	O
activate	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
,	NN	O	O
one	NN	O	O
through	NN	O	O
interactions	NN	O	O
with	NN	O	O
TRAF	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
the	NN	O	O
other	NN	O	O
with	NN	O	O
the	NN	O	O
TRADD	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
purpose	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
induction	NN	O	O
in	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
B	NN	O	O
cell	NN	O	O
activation	NN	O	O
markers	NN	O	O
ICAM-1	NN	O	B-protein
and	NN	O	O
CD71	NN	O	B-protein
by	NN	O	O
LMP1	NN	O	B-protein
.	NN	O	O

This	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
activates	NN	O	O
transcription	NN	O	O
from	NN	O	O
p50/p65-	NN	O	B-DNA
and	NN	O	I-DNA
c-Rel-	NN	O	I-DNA
responsive	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
activity	NN	O	O
can	NN	O	O
be	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
by	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
dominant	NN	O	O
inhibitory	NN	O	B-protein
IkappaB	NN	O	I-protein
mutant	NN	O	I-protein
.	NN	O	O

ICAM-1	NN	O	B-protein
and	NN	O	O
CD71	NN	O	B-protein
are	NN	O	O
nevertheless	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
LMP1	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
cell	NN	O	O
lines	NN	O	O
expressing	NN	O	O
the	NN	O	O
dominant	NN	O	B-protein
IkappaB	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
LMP1	NN	O	B-protein
-induced	NN	O	O
cell	NN	O	O
size	NN	O	O
increase	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
IkappaB	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
concluded	NN	O	O
that	NN	O	O
even	NN	O	O
when	NN	O	O
LMP1	NN	O	B-protein
is	NN	O	O
unable	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
it	NN	O	O
is	NN	O	O
still	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
certain	NN	O	O
characteristics	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
strongly	NN	O	O
suggesting	NN	O	O
that	NN	O	O
LMP1	NN	O	B-protein
can	NN	O	O
also	NN	O	O
activate	NN	O	O
cells	NN	O	O
independently	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Models	NN	O	O
of	NN	O	O
lineage	NN	O	O
switching	NN	O	O
in	NN	O	O
hematopoietic	NN	O	O
development	NN	O	O
:	NN	O	O
a	NN	O	O
new	NN	O	O
myeloid-committed	NN	O	B-cell_line
eosinophil	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
YJ	NN	O	B-cell_line
)	NN	O	O
demonstrates	NN	O	O
trilineage	NN	O	O
potential	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
human	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
with	NN	O	O
an	NN	O	O
eosinophilic	NN	O	O
phenotype	NN	O	O
,	NN	O	O
designated	NN	O	O
YJ	NN	O	B-cell_line
,	NN	O	O
was	NN	O	O
established	NN	O	O
from	NN	O	O
the	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
chronic	NN	O	O
myelomonocytic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
CMMoL	NN	O	O
)	NN	O	O
with	NN	O	O
eosinophilia	NN	O	O
.	NN	O	O

When	NN	O	O
cultured	NN	O	O
in	NN	O	O
RPMI	NN	O	O
1640	NN	O	O
medium	NN	O	O
containing	NN	O	O
10	NN	O	O
%	NN	O	O
fetal	NN	O	O
bovine	NN	O	O
serum	NN	O	O
,	NN	O	O
most	NN	O	O
YJ	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
myeloblastoid	NN	O	O
with	NN	O	O
a	NN	O	O
small	NN	O	O
number	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
having	NN	O	O
eosinophilic	NN	O	O
granules	NN	O	O
.	NN	O	O

Cell	NN	O	B-protein
surface	NN	O	I-protein
markers	NN	O	I-protein
in	NN	O	O
the	NN	O	O
YJ	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
positive	NN	O	O
for	NN	O	O
CD33	NN	O	B-protein
and	NN	O	O
were	NN	O	O
negative	NN	O	O
for	NN	O	O
CD34	NN	O	B-protein
,	NN	O	O
CD16	NN	O	B-protein
and	NN	O	O
CD23	NN	O	B-protein
.	NN	O	O

The	NN	O	O
eosinophilic	NN	O	O
characteristics	NN	O	O
of	NN	O	O
YJ	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
confirmed	NN	O	O
by	NN	O	O
histochemical	NN	O	O
staining	NN	O	O
with	NN	O	O
Fast-Green/Neutral-Red	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
mRNAs	NN	O	B-RNA
for	NN	O	O
eosinophil-associated	NN	O	B-protein
granule	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
eosinophil	NN	O	B-protein
cationic	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
ECP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
eosinophil-derived	NN	O	B-protein
neurotoxin	NN	O	I-protein
(	NN	O	O
EDN	NN	O	B-protein
)	NN	O	O
,	NN	O	O
eosinophil	NN	O	B-protein
peroxidase	NN	O	I-protein
(	NN	O	O
EPO	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
major	NN	O	B-protein
basic	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
MBP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
for	NN	O	O
the	NN	O	O
Charcot-Leyden	NN	O	B-protein
crystal	NN	O	I-protein
(	NN	O	I-protein
CLC	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
YJ	NN	O	B-cell_line
cells	NN	O	I-cell_line
could	NN	O	O
be	NN	O	O
induced	NN	O	O
towards	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
monocytic	NN	O	O
characteristics	NN	O	O
of	NN	O	O
YJ	NN	O	B-cell_line
cells	NN	O	I-cell_line
treated	NN	O	O
with	NN	O	O
PMA	NN	O	O
were	NN	O	O
confirmed	NN	O	O
by	NN	O	O
morphological	NN	O	O
analysis	NN	O	O
with	NN	O	O
alpha-naphthyl	NN	O	O
butyrate	NN	O	O
esterase	NN	O	O
staining	NN	O	O
,	NN	O	O
by	NN	O	O
CD14	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
by	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
YJ	NN	O	B-cell_line
cells	NN	O	I-cell_line
could	NN	O	O
be	NN	O	O
differentiated	NN	O	O
towards	NN	O	O
the	NN	O	O
neutrophil	NN	O	B-cell_type
lineage	NN	O	I-cell_type
by	NN	O	O
stimulation	NN	O	O
with	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
.	NN	O	O

YJ	NN	O	B-cell_line
cells	NN	O	I-cell_line
treated	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
2	NN	O	O
microM	NN	O	O
RA	NN	O	O
differentiated	NN	O	O
into	NN	O	O
metamyelocytes	NN	O	B-cell_type
and	NN	O	O
band	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
increased	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
nitroblue	NN	O	B-cell_line
tetrazolium	NN	O	I-cell_line
(	NN	O	I-cell_line
NBT	NN	O	I-cell_line
)	NN	O	I-cell_line
-positive	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
increased	NN	O	O
gp91phox	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
YJ	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
exhibited	NN	O	O
eosinophilic	NN	O	O
characteristics	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
differentiate	NN	O	O
to	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_type
or	NN	O	I-cell_type
neutrophilic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
in	NN	O	O
response	NN	O	O
to	NN	O	O
PMA	NN	O	O
or	NN	O	O
RA	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
for	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
was	NN	O	O
evaluated	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

Increased	NN	O	O
expression	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
with	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
increased	NN	O	O
expressions	NN	O	O
of	NN	O	O
C/EBPbeta	NN	O	B-protein
and	NN	O	O
MZF-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
occurred	NN	O	O
with	NN	O	O
neutrophilic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
YJ	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
should	NN	O	O
be	NN	O	O
useful	NN	O	O
for	NN	O	O
elucidating	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
governing	NN	O	O
lineage	NN	O	O
switching	NN	O	O
from	NN	O	O
the	NN	O	O
eosinophil	NN	O	B-cell_type
to	NN	O	O
monocytic	NN	O	B-cell_type
or	NN	O	I-cell_type
neutrophil	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Interleukin-6	NN	O	B-protein
production	NN	O	O
in	NN	O	O
hemorrhagic	NN	O	O
shock	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
neutrophil	NN	O	O
recruitment	NN	O	O
and	NN	O	O
lung	NN	O	O
injury	NN	O	O
.	NN	O	O

Hemorrhagic	NN	O	O
shock	NN	O	O
(	NN	O	O
HS	NN	O	O
)	NN	O	O
initiates	NN	O	O
an	NN	O	O
inflammatory	NN	O	O
cascade	NN	O	O
that	NN	O	O
includes	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
recruitment	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
(	NN	O	O
PMN	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
may	NN	O	O
progress	NN	O	O
to	NN	O	O
organ	NN	O	O
failure	NN	O	O
,	NN	O	O
inducing	NN	O	O
acute	NN	O	O
respiratory	NN	O	O
distress	NN	O	O
syndrome	NN	O	O
(	NN	O	O
ARDS	NN	O	O
)	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
interleukin-6	NN	O	B-protein
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
contributes	NN	O	O
to	NN	O	O
PMN	NN	O	O
infiltration	NN	O	O
and	NN	O	O
lung	NN	O	O
damage	NN	O	O
in	NN	O	O
HS	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
lungs	NN	O	O
of	NN	O	O
rats	NN	O	O
subjected	NN	O	O
to	NN	O	O
unresuscitated	NN	O	O
and	NN	O	O
resuscitated	NN	O	O
HS	NN	O	O
for	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat3	NN	O	B-protein
.	NN	O	O

Using	NN	O	O
semiquantitative	NN	O	O
RT-PCR	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
a	NN	O	O
striking	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
only	NN	O	O
in	NN	O	O
resuscitated	NN	O	O
HS	NN	O	O
,	NN	O	O
with	NN	O	O
peak	NN	O	O
levels	NN	O	O
observed	NN	O	O
1	NN	O	O
h	NN	O	O
after	NN	O	O
initiation	NN	O	O
of	NN	O	O
resuscitation	NN	O	O
.	NN	O	O

Increased	NN	O	O
IL-6	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
was	NN	O	O
localized	NN	O	O
to	NN	O	O
bronchial	NN	O	B-cell_type
and	NN	O	I-cell_type
alveolar	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
of	NN	O	O
protein	NN	O	O
extracts	NN	O	O
from	NN	O	O
shock	NN	O	O
lungs	NN	O	O
exhibited	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
Stat3	NN	O	B-protein
activation	NN	O	O
with	NN	O	O
kinetics	NN	O	O
similar	NN	O	O
to	NN	O	O
IL-6	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
situ	NN	O	O
DNA	NN	O	O
binding	NN	O	O
assay	NN	O	O
determined	NN	O	O
Stat3	NN	O	B-protein
activation	NN	O	O
predominantly	NN	O	O
within	NN	O	O
alveoli	NN	O	O
.	NN	O	O

Intratracheal	NN	O	O
instillation	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
alone	NN	O	O
into	NN	O	O
normal	NN	O	O
rats	NN	O	O
resulted	NN	O	O
in	NN	O	O
PMN	NN	O	O
infiltration	NN	O	O
into	NN	O	O
lung	NN	O	O
interstitium	NN	O	O
and	NN	O	O
alveoli	NN	O	O
,	NN	O	O
marked	NN	O	O
elevation	NN	O	O
of	NN	O	O
bronchoalveolar	NN	O	O
lavage	NN	O	O
cellularity	NN	O	O
,	NN	O	O
and	NN	O	O
increased	NN	O	O
wet-to-dry	NN	O	O
ratio	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-6	NN	O	B-protein
production	NN	O	O
and	NN	O	O
Stat3	NN	O	B-protein
activation	NN	O	O
occur	NN	O	O
early	NN	O	O
in	NN	O	O
HS	NN	O	O
and	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
PMN-mediated	NN	O	O
lung	NN	O	O
injury	NN	O	O
,	NN	O	O
including	NN	O	O
ARDS	NN	O	O
after	NN	O	O
HS	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
requires	NN	O	O
conserved	NN	O	O
leucine	NN	O	B-protein
charged	NN	O	I-protein
domains	NN	O	I-protein
for	NN	O	O
interactions	NN	O	O
with	NN	O	O
the	NN	O	O
conserved	NN	O	O
W	NN	O	B-DNA
box	NN	O	I-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
CIITA	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
(	NN	O	I-DNA
MHC	NN	O	I-DNA
)	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Aside	NN	O	O
from	NN	O	O
an	NN	O	O
N-terminal	NN	O	B-DNA
acidic	NN	O	I-DNA
transcriptional	NN	O	I-DNA
activation	NN	O	I-DNA
domain	NN	O	I-DNA
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
how	NN	O	O
this	NN	O	O
factor	NN	O	O
functions	NN	O	O
.	NN	O	O

Extensive	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
was	NN	O	O
undertaken	NN	O	O
to	NN	O	O
identify	NN	O	O
structural	NN	O	O
motifs	NN	O	O
required	NN	O	O
for	NN	O	O
function	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
mutants	NN	O	O
to	NN	O	O
activate	NN	O	O
a	NN	O	O
reporter	NN	O	O
gene	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
conserved	NN	O	O
W-X-Y	NN	O	B-DNA
or	NN	O	I-DNA
X-Y	NN	O	I-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
was	NN	O	O
determined	NN	O	O
.	NN	O	O

Two	NN	O	O
mutants	NN	O	O
displayed	NN	O	O
differential	NN	O	O
activity	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
promoters	NN	O	B-DNA
,	NN	O	O
activating	NN	O	O
transcription	NN	O	O
with	NN	O	O
the	NN	O	O
W-X-Y	NN	O	B-DNA
but	NN	O	I-DNA
not	NN	O	I-DNA
the	NN	O	I-DNA
X-Y	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

All	NN	O	O
mutants	NN	O	O
were	NN	O	O
tested	NN	O	O
for	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
interfere	NN	O	O
with	NN	O	O
wild-type	NN	O	O
CIITA	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Five	NN	O	O
CIITA	NN	O	B-DNA
mutant	NN	O	I-DNA
constructions	NN	O	I-DNA
were	NN	O	O
able	NN	O	O
to	NN	O	O
down-regulate	NN	O	O
wild-type	NN	O	O
CIITA	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Three	NN	O	O
of	NN	O	O
these	NN	O	O
mutants	NN	O	O
contained	NN	O	O
targeted	NN	O	O
disruptions	NN	O	O
of	NN	O	O
potential	NN	O	O
functional	NN	O	O
motifs	NN	O	O
:	NN	O	O
the	NN	O	O
acidic	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
a	NN	O	O
putative	NN	O	O
GTP-binding	NN	O	B-protein
motif	NN	O	I-protein
and	NN	O	O
two	NN	O	O
leucine	NN	O	B-protein
charged	NN	O	I-protein
domains	NN	O	I-protein
(	NN	O	O
LCD	NN	O	B-protein
motifs	NN	O	I-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
two	NN	O	O
contained	NN	O	O
mutations	NN	O	O
in	NN	O	O
regions	NN	O	O
that	NN	O	O
do	NN	O	O
not	NN	O	O
have	NN	O	O
homology	NN	O	O
to	NN	O	O
described	NN	O	O
proteins	NN	O	O
.	NN	O	O

The	NN	O	O
characterization	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
mutants	NN	O	I-protein
that	NN	O	O
are	NN	O	O
able	NN	O	O
to	NN	O	O
discriminate	NN	O	O
between	NN	O	O
promoters	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
the	NN	O	O
W	NN	O	B-DNA
box	NN	O	I-DNA
strongly	NN	O	O
suggests	NN	O	O
that	NN	O	O
CIITA	NN	O	B-protein
requires	NN	O	O
such	NN	O	O
interactions	NN	O	O
for	NN	O	O
function	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
LCD	NN	O	B-protein
motifs	NN	O	I-protein
required	NN	O	O
for	NN	O	O
CIITA	NN	O	B-protein
function	NN	O	O
brings	NN	O	O
to	NN	O	O
light	NN	O	O
a	NN	O	O
previously	NN	O	O
undefined	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
motifs	NN	O	O
in	NN	O	O
CIITA	NN	O	B-protein
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
PPARalpha	NN	O	B-protein
)	NN	O	O
ligand	NN	O	O
WY	NN	O	O
14	NN	O	O
,	NN	O	O
643	NN	O	O
does	NN	O	O
not	NN	O	O
interfere	NN	O	O
with	NN	O	O
leukotriene	NN	O	O
B4	NN	O	O
induced	NN	O	O
adhesion	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
PPAR	NN	O	B-protein
)	NN	O	O
control	NN	O	O
discrete	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
fatty	NN	O	O
acid	NN	O	O
and	NN	O	O
lipid	NN	O	O
metabolism	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
suggested	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
isoform	NN	O	I-protein
of	NN	O	O
PPAR	NN	O	B-protein
by	NN	O	O
the	NN	O	O
potent	NN	O	O
proinflammatory	NN	O	O
mediator	NN	O	O
leukotriene	NN	O	O
B4	NN	O	O
(	NN	O	O
LTB4	NN	O	O
)	NN	O	O
enhanced	NN	O	O
degradation	NN	O	O
of	NN	O	O
this	NN	O	O
eicosanoid	NN	O	O
,	NN	O	O
offersuggesting	NN	O	O
a	NN	O	O
new	NN	O	O
aspect	NN	O	O
of	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
whether	NN	O	O
PPARalpha	NN	O	B-protein
activation	NN	O	O
(	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
the	NN	O	O
selective	NN	O	O
agonist	NN	O	O
WY	NN	O	O
14	NN	O	O
,	NN	O	O
643	NN	O	O
)	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
pivotal	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
,	NN	O	O
interfered	NN	O	O
with	NN	O	O
LTB4	NN	O	O
induced	NN	O	O
adhesion	NN	O	O
of	NN	O	O
PMN	NN	O	B-cell_type
neutrophil	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

When	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
treated	NN	O	O
with	NN	O	O
WY	NN	O	O
14	NN	O	O
,	NN	O	O
643	NN	O	O
prior	NN	O	O
to	NN	O	O
activation	NN	O	O
with	NN	O	O
LTB4	NN	O	O
(	NN	O	O
or	NN	O	O
fMLP	NN	O	B-protein
,	NN	O	O
IL-1beta	NN	O	B-protein
or	NN	O	O
TNFalpha	NN	O	B-protein
,	NN	O	O
as	NN	O	O
controls	NN	O	O
)	NN	O	O
we	NN	O	O
could	NN	O	O
not	NN	O	O
document	NN	O	O
any	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
adhering	NN	O	B-cell_type
PMN	NN	O	I-cell_type
or	NN	O	O
duration	NN	O	O
of	NN	O	O
the	NN	O	O
response	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
this	NN	O	O
study	NN	O	O
provides	NN	O	O
no	NN	O	O
evidence	NN	O	O
indicating	NN	O	O
a	NN	O	O
regulatory	NN	O	O
function	NN	O	O
of	NN	O	O
PPARalpha	NN	O	B-protein
in	NN	O	O
LTB4	NN	O	O
induced	NN	O	O
adhesive	NN	O	O
interactions	NN	O	O
between	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
neutrophils	NN	O	B-cell_type

-DOCSTART-	O

Interaction	NN	O	O
of	NN	O	O
sickle	NN	O	B-cell_type
erythrocytes	NN	O	I-cell_type
with	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
endothelial	NN	O	O
cell	NN	O	O
conditioned	NN	O	O
medium	NN	O	O
induces	NN	O	O
oxidant	NN	O	O
stress	NN	O	O
leading	NN	O	O
to	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
abnormal	NN	O	O
adherence	NN	O	O
of	NN	O	O
sickle	NN	O	B-cell_type
red	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
SS	NN	O	B-cell_type
RBC	NN	O	I-cell_type
)	NN	O	O
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
thought	NN	O	O
to	NN	O	O
contribute	NN	O	O
to	NN	O	O
vascular	NN	O	O
occlusion	NN	O	O
,	NN	O	O
a	NN	O	O
major	NN	O	O
cause	NN	O	O
of	NN	O	O
morbidity	NN	O	O
in	NN	O	O
sickle	NN	O	O
cell	NN	O	O
disease	NN	O	O
(	NN	O	O
SCD	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
determined	NN	O	O
whether	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
SS	NN	O	B-cell_type
RBC	NN	O	I-cell_type
with	NN	O	O
cultured	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
induced	NN	O	O
cellular	NN	O	O
oxidant	NN	O	O
stress	NN	O	O
that	NN	O	O
would	NN	O	O
culminate	NN	O	O
in	NN	O	O
expression	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
(	NN	O	O
CAMs	NN	O	B-protein
)	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
adhesion	NN	O	O
and	NN	O	O
diapedesis	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
the	NN	O	O
adherence	NN	O	O
of	NN	O	O
SS	NN	O	B-cell_type
reticulocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
SS	NN	O	B-cell_type
RBC	NN	O	I-cell_type
at	NN	O	O
2	NN	O	O
%	NN	O	O
concentration	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
multimers	NN	O	O
of	NN	O	O
von	NN	O	B-protein
Willebrand	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
vWf	NN	O	B-protein
)	NN	O	O
,	NN	O	O
derived	NN	O	O
from	NN	O	O
endothelial	NN	O	O
cell-derived	NN	O	O
conditioned	NN	O	O
medium	NN	O	O
(	NN	O	O
E-CM	NN	O	O
)	NN	O	O
with	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
umbilical	NN	O	I-cell_line
vein	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
HUVEC	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
fivefold	NN	O	O
increased	NN	O	O
formation	NN	O	O
of	NN	O	O
thiobarbituric	NN	O	O
acid-reactive	NN	O	O
substances	NN	O	O
(	NN	O	O
TBARS	NN	O	O
)	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kB	NN	O	B-protein
,	NN	O	O
both	NN	O	O
indicators	NN	O	O
of	NN	O	O
cellular	NN	O	O
oxidant	NN	O	O
stress	NN	O	O
.	NN	O	O

Normal	NN	O	B-cell_type
RBC	NN	O	I-cell_type
show	NN	O	O
none	NN	O	O
of	NN	O	O
these	NN	O	O
phenomena	NN	O	O
.	NN	O	O

The	NN	O	O
oxidant	NN	O	O
stress-induced	NN	O	O
signaling	NN	O	O
resulted	NN	O	O
in	NN	O	O
an	NN	O	O
increased	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
CAMs	NN	O	B-protein
,	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
VCAM-1	NN	O	B-protein
in	NN	O	O
HUVEC	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
oxygen	NN	O	B-protein
radical	NN	O	I-protein
scavenger	NN	O	I-protein
enzymes	NN	O	I-protein
(	NN	O	O
catalase	NN	O	B-protein
,	NN	O	O
superoxide	NN	O	B-protein
dismutase	NN	O	I-protein
)	NN	O	O
and	NN	O	O
antioxidant	NN	O	O
(	NN	O	O
probucol	NN	O	O
)	NN	O	O
inhibited	NN	O	O
these	NN	O	O
events	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
preincubation	NN	O	O
of	NN	O	O
HUVEC	NN	O	B-cell_line
with	NN	O	O
a	NN	O	O
synthetic	NN	O	O
peptide	NN	O	O
Arg-Gly-Asp	NN	O	O
(	NN	O	O
RGD	NN	O	O
)	NN	O	O
that	NN	O	O
prevents	NN	O	O
vWf	NN	O	B-protein
-mediated	NN	O	O
adhesion	NN	O	O
of	NN	O	O
SS	NN	O	B-cell_type
RBC	NN	O	I-cell_type
reduced	NN	O	O
the	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
VCAM-1	NN	O	B-protein
and	NN	O	O
NF-kB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
SS	NN	O	B-cell_type
RBC	NN	O	I-cell_type
-induced	NN	O	O
oxidant	NN	O	O
stress	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
twofold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
transendothelial	NN	O	O
migration	NN	O	O
of	NN	O	O
both	NN	O	O
monocyte-like	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
approximately	NN	O	O
a	NN	O	O
sixfold	NN	O	O
increase	NN	O	O
in	NN	O	O
platelet-endothelial	NN	O	B-protein
cell	NN	O	I-protein
adhesion	NN	O	I-protein
molecule-1	NN	O	I-protein
(	NN	O	O
PECAM-1	NN	O	B-protein
)	NN	O	O
phosphorylation	NN	O	O
,	NN	O	O
each	NN	O	O
of	NN	O	O
which	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
inhibitor	NN	O	O
and	NN	O	O
antioxidants	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
adherence/contact	NN	O	O
of	NN	O	O
SS	NN	O	B-cell_type
RBC	NN	O	I-cell_type
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
large	NN	O	O
vessel	NN	O	O
can	NN	O	O
generate	NN	O	O
enhanced	NN	O	O
oxidant	NN	O	O
stress	NN	O	O
leading	NN	O	O
to	NN	O	O
increased	NN	O	O
adhesion	NN	O	O
and	NN	O	O
diapedesis	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
heightened	NN	O	O
adherence	NN	O	O
of	NN	O	O
SS	NN	O	B-cell_type
reticulocytes	NN	O	I-cell_type
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
injury/activation	NN	O	O
of	NN	O	O
endothelium	NN	O	O
can	NN	O	O
contribute	NN	O	O
to	NN	O	O
vaso-occlusion	NN	O	O
in	NN	O	O
SCD	NN	O	O
.	NN	O	O

-DOCSTART-	O

Signal	NN	O	O
transduction	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
advanced	NN	O	O
renal	NN	O	O
carcinoma	NN	O	O
:	NN	O	O
clinical	NN	O	O
relevance	NN	O	O
and	NN	O	O
effects	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
therapy	NN	O	O
.	NN	O	O

Studies	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
the	NN	O	O
CD3/T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
pathways	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
animals	NN	O	O
and	NN	O	O
patients	NN	O	O
with	NN	O	O
advanced	NN	O	O
malignancy	NN	O	O
.	NN	O	O

Diminished	NN	O	O
expression	NN	O	O
of	NN	O	O
TCRzeta	NN	O	B-protein
and	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
that	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
reduced	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
RelA	NN	O	B-protein
containing	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	I-protein
NFkappaB	NN	O	I-protein
)	NN	O	I-protein
complexes	NN	O	I-protein
have	NN	O	O
been	NN	O	O
noted	NN	O	O
.	NN	O	O

These	NN	O	O
defects	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
malignant	NN	O	O
melanoma	NN	O	O
,	NN	O	O
renal	NN	O	O
cell	NN	O	O
carcinoma	NN	O	O
(	NN	O	O
RCC	NN	O	O
)	NN	O	O
,	NN	O	O
ovarian	NN	O	O
cancer	NN	O	O
,	NN	O	O
and	NN	O	O
colorectal	NN	O	O
cancer	NN	O	O
.	NN	O	O

Preliminary	NN	O	O
observations	NN	O	O
also	NN	O	O
indicate	NN	O	O
possible	NN	O	O
correlation	NN	O	O
with	NN	O	O
clinical	NN	O	O
variables	NN	O	O
such	NN	O	O
as	NN	O	O
stage	NN	O	O
in	NN	O	O
selected	NN	O	O
instances	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
characterize	NN	O	O
altered	NN	O	O
expression	NN	O	O
of	NN	O	O
TCRzeta	NN	O	B-protein
,	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
and	NN	O	O
impaired	NN	O	O
activation	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
,	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
obtained	NN	O	O
from	NN	O	O
65	NN	O	O
patients	NN	O	O
with	NN	O	O
RCC	NN	O	O
,	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
whom	NN	O	O
were	NN	O	O
receiving	NN	O	O
combination	NN	O	O
cytokine	NN	O	B-protein
therapy	NN	O	O
[	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
,	NN	O	O
IFN	NN	O	B-protein
alpha	NN	O	I-protein
-containing	NN	O	O
regimens	NN	O	O
]	NN	O	O
and	NN	O	O
37	NN	O	O
control	NN	O	O
individuals	NN	O	O
.	NN	O	O

In	NN	O	O
29	NN	O	O
of	NN	O	O
these	NN	O	O
patients	NN	O	O
,	NN	O	O
levels	NN	O	O
of	NN	O	O
TCRzeta	NN	O	B-protein
and	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
were	NN	O	O
determined	NN	O	O
by	NN	O	O
Western	NN	O	O
blots	NN	O	O
of	NN	O	O
T-cell	NN	O	O
lysates	NN	O	O
and	NN	O	O
semiquantitated	NN	O	O
using	NN	O	O
densitometry	NN	O	O
.	NN	O	O

Relative	NN	O	O
levels	NN	O	O
were	NN	O	O
then	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
clinical	NN	O	O
variables	NN	O	O
including	NN	O	O
response	NN	O	O
to	NN	O	O
therapy	NN	O	O
,	NN	O	O
performance	NN	O	O
status	NN	O	O
,	NN	O	O
survival	NN	O	O
,	NN	O	O
disease	NN	O	O
sites	NN	O	O
,	NN	O	O
age	NN	O	O
,	NN	O	O
and	NN	O	O
others	NN	O	O
.	NN	O	O

In	NN	O	O
another	NN	O	O
group	NN	O	O
of	NN	O	O
28	NN	O	O
patients	NN	O	O
(	NN	O	O
three	NN	O	O
individuals	NN	O	O
from	NN	O	O
the	NN	O	O
first	NN	O	O
group	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
abnormal	NN	O	O
NFkappaB	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
studied	NN	O	O
using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
after	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate/ionomycin	NN	O	O
or	NN	O	O
anti-CD3	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
.	NN	O	O

Changes	NN	O	O
in	NN	O	O
these	NN	O	O
signaling	NN	O	O
molecules	NN	O	O
during	NN	O	O
cytokine	NN	O	B-protein
treatment	NN	O	O
were	NN	O	O
also	NN	O	O
investigated	NN	O	O
.	NN	O	O

TCRzeta	NN	O	B-protein
and	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
27	NN	O	O
of	NN	O	O
29	NN	O	O
patients	NN	O	O
,	NN	O	O
and	NN	O	O
overall	NN	O	O
,	NN	O	O
reduced	NN	O	O
levels	NN	O	O
were	NN	O	O
noted	NN	O	O
visually	NN	O	O
in	NN	O	O
12	NN	O	O
of	NN	O	O
29	NN	O	O
(	NN	O	O
41	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
13	NN	O	O
of	NN	O	O
29	NN	O	O
(	NN	O	O
45	NN	O	O
%	NN	O	O
)	NN	O	O
individuals	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

When	NN	O	O
levels	NN	O	O
were	NN	O	O
semiquantitated	NN	O	O
using	NN	O	O
densitometry	NN	O	O
,	NN	O	O
significant	NN	O	O
decreases	NN	O	O
of	NN	O	O
TCRzeta	NN	O	B-protein
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.029	NN	O	O
)	NN	O	O
and	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.029	NN	O	O
)	NN	O	O
but	NN	O	O
not	NN	O	O
CD3epsilon	NN	O	B-protein
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.131	NN	O	O
)	NN	O	O
,	NN	O	O
compared	NN	O	O
with	NN	O	O
control	NN	O	O
levels	NN	O	O
,	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

In	NN	O	O
patients	NN	O	O
treated	NN	O	O
with	NN	O	O
IL-2/	NN	O	O
IFN	NN	O	B-protein
alpha	NN	O	I-protein
-based	NN	O	O
therapy	NN	O	O
,	NN	O	O
relative	NN	O	O
levels	NN	O	O
of	NN	O	O
TCRzeta	NN	O	B-protein
increased	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.002	NN	O	O
)	NN	O	O
on	NN	O	O
day	NN	O	O
15	NN	O	O
of	NN	O	O
cycle	NN	O	O
one	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
baseline	NN	O	O
.	NN	O	O

Correlations	NN	O	O
of	NN	O	O
TCRzeta	NN	O	B-protein
or	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
levels	NN	O	O
with	NN	O	O
response	NN	O	O
or	NN	O	O
disease	NN	O	O
variables	NN	O	O
,	NN	O	O
except	NN	O	O
for	NN	O	O
lower	NN	O	O
TCRzeta	NN	O	B-protein
levels	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
bone	NN	O	O
metastases	NN	O	O
,	NN	O	O
were	NN	O	O
not	NN	O	O
found	NN	O	O
.	NN	O	O

Abnormal	NN	O	O
NFkappaB	NN	O	B-protein
activation	NN	O	O
after	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate/ionomycin	NN	O	O
and/or	NN	O	O
anti-CD3	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
was	NN	O	O
found	NN	O	O
in	NN	O	O
59	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
(	NN	O	O
17	NN	O	O
of	NN	O	O
28	NN	O	O
)	NN	O	O
and	NN	O	O
was	NN	O	O
not	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
the	NN	O	O
advanced	NN	O	O
age	NN	O	O
of	NN	O	O
the	NN	O	O
study	NN	O	O
cohort	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
inducible	NN	O	O
during	NN	O	O
cytokine	NN	O	B-protein
therapy	NN	O	O
in	NN	O	O
four	NN	O	O
of	NN	O	O
six	NN	O	O
individuals	NN	O	O
who	NN	O	O
displayed	NN	O	O
impaired	NN	O	O
NFkappaB	NN	O	B-protein
activity	NN	O	O
prior	NN	O	O
to	NN	O	O
therapy	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
impaired	NN	O	O
activation	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
does	NN	O	O
not	NN	O	O
appear	NN	O	O
linked	NN	O	O
to	NN	O	O
a	NN	O	O
reduction	NN	O	O
of	NN	O	O
TCRzeta	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
because	NN	O	O
in	NN	O	O
five	NN	O	O
patients	NN	O	O
,	NN	O	O
normal	NN	O	O
TCRzeta	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
present	NN	O	O
although	NN	O	O
kappaB	NN	O	B-protein
binding	NN	O	O
was	NN	O	O
not	NN	O	O
inducible	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
advanced	NN	O	O
RCC	NN	O	O
,	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
TCRzeta	NN	O	B-protein
and	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
subset	NN	O	O
,	NN	O	O
reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
these	NN	O	O
TCRzeta	NN	O	B-protein
associated	NN	O	I-protein
molecules	NN	O	I-protein
are	NN	O	O
seen	NN	O	O
that	NN	O	O
may	NN	O	O
increase	NN	O	O
during	NN	O	O
cytokine	NN	O	B-protein
-based	NN	O	O
therapy	NN	O	O
.	NN	O	O

Abnormal	NN	O	O
activation	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
is	NN	O	O
also	NN	O	O
present	NN	O	O
in	NN	O	O
>	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
and	NN	O	O
may	NN	O	O
also	NN	O	O
revert	NN	O	O
to	NN	O	O
normal	NN	O	O
during	NN	O	O
IL-2/	NN	O	O
IFN	NN	O	B-protein
alpha	NN	O	I-protein
-based	NN	O	O
treatment	NN	O	O
.	NN	O	O

This	NN	O	O
alteration	NN	O	O
in	NN	O	O
NFkappaB	NN	O	B-protein
activation	NN	O	O
occurred	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
normal	NN	O	O
expression	NN	O	O
of	NN	O	O
TCRzeta-associated	NN	O	B-DNA
signaling	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
significance	NN	O	O
of	NN	O	O
these	NN	O	O
findings	NN	O	O
remains	NN	O	O
unclear	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
expressed	NN	O	O
during	NN	O	O
ATRA-induced	NN	O	O
neutrophil	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
recent	NN	O	O
clinical	NN	O	O
therapeutic	NN	O	O
initiative	NN	O	O
has	NN	O	O
been	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
chemical	NN	O	O
agents	NN	O	O
which	NN	O	O
induce	NN	O	O
the	NN	O	O
leukaemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
overcome	NN	O	O
their	NN	O	O
block	NN	O	O
in	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
understand	NN	O	O
this	NN	O	O
block	NN	O	O
the	NN	O	O
cascade	NN	O	O
of	NN	O	O
molecular	NN	O	O
events	NN	O	O
needs	NN	O	O
to	NN	O	O
be	NN	O	O
characterized	NN	O	O
.	NN	O	O

Haemopoietic	NN	O	O
differentiation	NN	O	O
is	NN	O	O
ultimately	NN	O	O
controlled	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
which	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
an	NN	O	O
array	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Many	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
contain	NN	O	O
similar	NN	O	O
structural	NN	O	B-protein
protein	NN	O	I-protein
sequences	NN	O	I-protein
,	NN	O	O
and	NN	O	O
we	NN	O	O
have	NN	O	O
used	NN	O	O
an	NN	O	O
RT-PCR-based	NN	O	O
approach	NN	O	O
to	NN	O	O
isolate	NN	O	O
sequences	NN	O	O
,	NN	O	O
from	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
gene	NN	O	O
families	NN	O	O
which	NN	O	O
share	NN	O	O
similar	NN	O	O
domains	NN	O	O
.	NN	O	O

Degenerate	NN	O	O
primers	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
TFIIIA	NN	O	B-DNA
zinc-finger	NN	O	I-DNA
consensus	NN	O	I-DNA
amino	NN	O	I-DNA
acid	NN	O	I-DNA
sequences	NN	O	I-DNA
and	NN	O	O
to	NN	O	O
the	NN	O	O
POU-homeodomain	NN	O	B-protein
and	NN	O	O
POU-specific	NN	O	B-protein
domain	NN	O	I-protein
were	NN	O	O
used	NN	O	O
to	NN	O	O
amplify	NN	O	O
genes	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
that	NN	O	O
they	NN	O	O
contained	NN	O	O
similarities	NN	O	O
in	NN	O	O
structural	NN	O	O
motifs	NN	O	O
shared	NN	O	O
within	NN	O	O
these	NN	O	O
families	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

A	NN	O	O
serum-independent	NN	O	B-cell_line
HL60	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
was	NN	O	O
induced	NN	O	O
towards	NN	O	O
the	NN	O	O
neutrophil	NN	O	O
lineage	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
all-trans	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
ATRA	NN	O	O
)	NN	O	O
for	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

CD38+	NN	O	B-cell_type
cells	NN	O	I-cell_type
committed	NN	O	O
towards	NN	O	O
this	NN	O	O
lineage	NN	O	O
were	NN	O	O
enriched	NN	O	O
and	NN	O	O
a	NN	O	O
population	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
treated	NN	O	O
with	NN	O	O
dihydroxyvitamin	NN	O	O
D3	NN	O	O
to	NN	O	O
induce	NN	O	O
neutrophil	NN	O	O
maturation	NN	O	O
.	NN	O	O

RNA	NN	O	O
extracted	NN	O	O
from	NN	O	O
uninduced	NN	O	O
,	NN	O	O
ATRA-induced	NN	O	B-cell_line
CD38+	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
vitamin	NN	O	B-cell_line
D3	NN	O	I-cell_line
treated	NN	O	I-cell_line
maturing	NN	O	I-cell_line
cell	NN	O	I-cell_line
cultures	NN	O	I-cell_line
were	NN	O	O
amplified	NN	O	O
using	NN	O	O
the	NN	O	O
degenerate	NN	O	O
primers	NN	O	O
.	NN	O	O

PCR	NN	O	B-DNA
fragments	NN	O	I-DNA
were	NN	O	O
cloned	NN	O	O
,	NN	O	O
sequenced	NN	O	O
,	NN	O	O
clustered	NN	O	O
into	NN	O	O
homologous	NN	O	O
groups	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
group	NN	O	O
sequences	NN	O	O
searched	NN	O	O
on	NN	O	O
the	NN	O	O
GenBank	NN	O	O
database	NN	O	O
.	NN	O	O

The	NN	O	O
Oct	NN	O	O
1	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
very	NN	O	O
close	NN	O	O
homologue	NN	O	O
,	NN	O	O
KIAA0144	NN	O	B-DNA
,	NN	O	O
was	NN	O	O
identified	NN	O	O
using	NN	O	O
the	NN	O	O
POU	NN	O	O
family	NN	O	O
primers	NN	O	O
.	NN	O	O

The	NN	O	O
zinc-finger	NN	O	O
primers	NN	O	O
identified	NN	O	O
three	NN	O	O
zinc-finger	NN	O	O
genes	NN	O	O
.	NN	O	O

The	NN	O	O
pattern	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
suggested	NN	O	O
from	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
clones	NN	O	O
in	NN	O	O
each	NN	O	O
group	NN	O	O
at	NN	O	O
neutrophil	NN	O	O
commitment	NN	O	O
and	NN	O	O
maturation	NN	O	O
.	NN	O	O

The	NN	O	O
differential	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
in	NN	O	O
the	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
and	NN	O	I-protein
POU	NN	O	I-protein
families	NN	O	I-protein
will	NN	O	O
lead	NN	O	O
to	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
cascade	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
which	NN	O	O
occurs	NN	O	O
following	NN	O	O
ATRA-induced	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

GATA-3	NN	O	B-protein
-dependent	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
in	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
regulation	NN	O	O
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
directing	NN	O	O
Th2-specific	NN	O	O
activity	NN	O	O
.	NN	O	O

An	NN	O	O
800-base	NN	O	B-DNA
pair	NN	O	I-DNA
proximal	NN	O	I-DNA
promoter	NN	O	I-DNA
conferred	NN	O	O
some	NN	O	O
Th2-selective	NN	O	O
expression	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
region	NN	O	O
directed	NN	O	O
extremely	NN	O	O
low	NN	O	O
reporter	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
relative	NN	O	O
to	NN	O	O
endogenous	NN	O	O
IL-4	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
full	NN	O	O
gene	NN	O	O
activity	NN	O	O
requires	NN	O	O
additional	NN	O	O
enhancer	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
large	NN	O	O
genomic	NN	O	B-DNA
IL-4	NN	O	I-DNA
regions	NN	O	I-DNA
for	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
and	NN	O	O
interaction	NN	O	O
with	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
proximal	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
only	NN	O	O
moderately	NN	O	O
augmented	NN	O	O
by	NN	O	O
GATA-3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
certain	NN	O	O
genomic	NN	O	B-DNA
regions	NN	O	I-DNA
significantly	NN	O	O
enhanced	NN	O	O
GATA-3	NN	O	B-DNA
promoter	NN	O	I-DNA
transactivation	NN	O	O
.	NN	O	O

Some	NN	O	O
enhancing	NN	O	O
regions	NN	O	O
contained	NN	O	O
consensus	NN	O	O
,	NN	O	O
GATA	NN	O	B-DNA
sites	NN	O	I-DNA
that	NN	O	O
bound	NN	O	O
Th2-specific	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
retroviral	NN	O	O
transduction	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
into	NN	O	O
developing	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
induced	NN	O	O
IL-5	NN	O	B-protein
to	NN	O	O
full	NN	O	O
Th2	NN	O	B-protein
levels	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
partially	NN	O	O
restored	NN	O	O
IL-4	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
GATA-3	NN	O	B-protein
is	NN	O	O
permissive	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
sufficient	NN	O	O
,	NN	O	O
for	NN	O	O
full	NN	O	O
IL-4	NN	O	B-protein
enhancement	NN	O	O
and	NN	O	O
may	NN	O	O
act	NN	O	O
through	NN	O	O
GATA	NN	O	B-DNA
elements	NN	O	I-DNA
surrounding	NN	O	O
the	NN	O	O
IL-13/IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
locus	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Position	NN	O	O
effect	NN	O	O
of	NN	O	O
translocations	NN	O	O
involving	NN	O	O
the	NN	O	O
inactive	NN	O	B-DNA
X	NN	O	I-DNA
chromosome	NN	O	I-DNA
:	NN	O	O
physical	NN	O	O
linkage	NN	O	O
to	NN	O	O
XIC/XIST	NN	O	B-DNA
does	NN	O	O
not	NN	O	O
lead	NN	O	O
to	NN	O	O
long-range	NN	O	O
de	NN	O	O
novo	NN	O	O
inactivation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
differentiated	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Given	NN	O	O
the	NN	O	O
reported	NN	O	O
long-range	NN	O	O
cis-inactivating	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
XIST	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
early	NN	O	O
embryonic	NN	O	O
development	NN	O	O
and	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
requirement	NN	O	O
of	NN	O	O
X-chromosome-specific	NN	O	B-DNA
elements	NN	O	I-DNA
for	NN	O	O
propagating	NN	O	O
the	NN	O	O
inactive	NN	O	O
state	NN	O	O
,	NN	O	O
there	NN	O	O
exists	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
cis	NN	O	O
inactivation	NN	O	O
of	NN	O	O
autosomal	NN	O	O
material	NN	O	O
after	NN	O	O
de	NN	O	O
novo	NN	O	O
translocation	NN	O	O
to	NN	O	O
an	NN	O	O
inactive	NN	O	B-DNA
X	NN	O	I-DNA
chromosome	NN	O	I-DNA
(	NN	O	O
Xi	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
differentiated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
de	NN	O	O
novo	NN	O	O
radiation-induced	NN	O	O
translocations	NN	O	O
between	NN	O	O
the	NN	O	O
Xi	NN	O	B-DNA
and	NN	O	O
autosomes	NN	O	B-DNA
to	NN	O	O
study	NN	O	O
the	NN	O	O
maintenance	NN	O	O
and	NN	O	O
spreading	NN	O	O
of	NN	O	O
X-chromosome	NN	O	B-DNA
inactivation	NN	O	O
(	NN	O	O
X	NN	O	O
inactivation	NN	O	O
)	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
X-inactivation	NN	O	O
center	NN	O	O
(	NN	O	B-DNA
XIC	NN	O	I-DNA
)	NN	O	I-DNA
/XIST	NN	O	I-DNA
in	NN	O	O
differentiated	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Autosome/	NN	O	O
Xi	NN	O	B-DNA
translocations	NN	O	O
were	NN	O	O
detected	NN	O	O
by	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
chromosomes	NN	O	B-DNA
involved	NN	O	O
in	NN	O	O
the	NN	O	O
translocation	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
simultaneous	NN	O	O
immunocytogenetic	NN	O	O
studies	NN	O	O
using	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
either	NN	O	O
BrdU	NN	O	B-protein
incorporated	NN	O	O
at	NN	O	O
late	NN	O	O
S	NN	O	O
phase	NN	O	O
or	NN	O	O
acetylated	NN	O	B-protein
histone	NN	O	I-protein
H4	NN	O	I-protein
.	NN	O	O

The	NN	O	O
position	NN	O	O
of	NN	O	O
XIC/XIST	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
reciprocal	NN	O	O
products	NN	O	O
of	NN	O	O
the	NN	O	O
translocation	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
XIST-specific	NN	O	O
FISH	NN	O	O
and	NN	O	O
computer	NN	O	O
enhancement	NN	O	O
.	NN	O	O

In	NN	O	O
other	NN	O	O
experiments	NN	O	O
,	NN	O	O
the	NN	O	O
Xq13	NN	O	B-DNA
region	NN	O	I-DNA
carrying	NN	O	O
XIC/XIST	NN	O	B-DNA
was	NN	O	O
localized	NN	O	O
by	NN	O	O
computer	NN	O	O
enhancement	NN	O	O
of	NN	O	O
the	NN	O	O
DAPI	NN	O	O
banding	NN	O	O
pattern	NN	O	O
.	NN	O	O

Our	NN	O	O
study	NN	O	O
in	NN	O	O
differentiated	NN	O	B-cell_type
cells	NN	O	I-cell_type
provides	NN	O	O
a	NN	O	O
visual	NN	O	O
demonstration	NN	O	O
that	NN	O	O
physical	NN	O	O
separation	NN	O	O
from	NN	O	O
XIC/XIST	NN	O	B-DNA
does	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
reactivation	NN	O	O
of	NN	O	O
inactive	NN	O	B-DNA
X-chromosome	NN	O	I-DNA
material	NN	O	O
and	NN	O	O
that	NN	O	O
X	NN	O	O
inactivation	NN	O	O
is	NN	O	O
not	NN	O	O
spread	NN	O	O
to	NN	O	O
the	NN	O	O
translocated	NN	O	B-DNA
autosomes	NN	O	I-DNA
irrespective	NN	O	O
of	NN	O	O
the	NN	O	O
position	NN	O	O
of	NN	O	O
XIC/XIST	NN	O	B-DNA
.	NN	O	O

This	NN	O	O
observation	NN	O	O
suggests	NN	O	O
that	NN	O	O
physical	NN	O	O
linkage	NN	O	O
to	NN	O	O
XIC/XIST	NN	O	B-DNA
does	NN	O	O
not	NN	O	O
lead	NN	O	O
to	NN	O	O
de	NN	O	O
novo	NN	O	O
inactivation	NN	O	O
of	NN	O	O
autosomal	NN	O	O
material	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin-4	NN	O	B-protein
and	NN	O	I-protein
-13	NN	O	I-protein
induce	NN	O	O
upregulation	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	O
macrophage	NN	O	O
12/15-lipoxygenase	NN	O	B-protein
activity	NN	O	I-protein
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
STAT6	NN	O	B-protein
.	NN	O	O

When	NN	O	O
human	NN	O	O
monocytes	NN	O	B-cell_type
or	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
are	NN	O	O
cultured	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-4	NN	O	I-protein
or	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
reticulocyte-type	NN	O	B-protein
15-lipoxygenase	NN	O	I-protein
is	NN	O	O
induced	NN	O	O
.	NN	O	O

In	NN	O	O
mice	NN	O	O
a	NN	O	O
15-lipoxygenase	NN	O	B-protein
is	NN	O	O
not	NN	O	O
expressed	NN	O	O
,	NN	O	O
but	NN	O	O
a	NN	O	O
leukocyte-type	NN	O	B-protein
12-lipoxygenase	NN	O	I-protein
is	NN	O	O
present	NN	O	O
in	NN	O	O
peritoneal	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
whether	NN	O	O
both	NN	O	O
lipoxygenase	NN	O	B-protein
isoforms	NN	O	I-protein
exhibit	NN	O	O
a	NN	O	O
similar	NN	O	O
regulatory	NN	O	O
response	NN	O	O
toward	NN	O	O
cytokine	NN	O	B-protein
stimulation	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
leukocyte-type	NN	O	B-protein
12-lipoxygenase	NN	O	I-protein
of	NN	O	O
murine	NN	O	B-cell_type
peritoneal	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
by	NN	O	O
interleukins	NN	O	B-protein
and	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
enzyme	NN	O	B-protein
is	NN	O	O
upregulated	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
when	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
cultured	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
or	NN	O	O
IL-13	NN	O	B-protein
but	NN	O	O
not	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
.	NN	O	O

When	NN	O	O
peripheral	NN	O	O
murine	NN	O	O
monocytes	NN	O	B-cell_type
that	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
the	NN	O	O
lipoxygenase	NN	O	B-protein
were	NN	O	O
treated	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
expression	NN	O	O
of	NN	O	O
12/15-lipoxygenase	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
induced	NN	O	O
,	NN	O	O
suggesting	NN	O	O
pretranslational	NN	O	O
control	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
no	NN	O	O
upregulation	NN	O	O
of	NN	O	O
the	NN	O	O
lipoxygenase	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
observed	NN	O	O
when	NN	O	O
the	NN	O	O
macrophages	NN	O	B-cell_type
were	NN	O	O
prepared	NN	O	O
from	NN	O	O
homozygous	NN	O	O
STAT6	NN	O	B-protein
-deficient	NN	O	O
mice	NN	O	O
.	NN	O	O

Peritoneal	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
of	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
that	NN	O	O
systemically	NN	O	O
overexpress	NN	O	O
IL-4	NN	O	B-protein
exhibited	NN	O	O
a	NN	O	O
threefold	NN	O	O
to	NN	O	O
fourfold	NN	O	O
higher	NN	O	O
12-lipoxygenase	NN	O	B-protein
activity	NN	O	O
than	NN	O	O
cells	NN	O	O
prepared	NN	O	O
from	NN	O	O
control	NN	O	O
animals	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
upregulation	NN	O	O
of	NN	O	O
12-lipoxygenase	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
heart	NN	O	O
,	NN	O	O
spleen	NN	O	O
,	NN	O	O
and	NN	O	O
lung	NN	O	O
of	NN	O	O
the	NN	O	O
transgenic	NN	O	O
animals	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
a	NN	O	O
strong	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
red	NN	O	B-cell_type
cells	NN	O	I-cell_type
during	NN	O	O
experimental	NN	O	O
anemia	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
presented	NN	O	O
here	NN	O	O
indicate	NN	O	O
that	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
12-lipoxygenase	NN	O	B-protein
activity	NN	O	O
of	NN	O	O
murine	NN	O	O
macrophages	NN	O	O
is	NN	O	O
upregulated	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
and/or	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
this	NN	O	O
upregulation	NN	O	O
requires	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
STAT6	NN	O	B-protein
,	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
the	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
enzyme	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
STAT6	NN	O	B-protein
independent	NN	O	O
.	NN	O	O

The	NN	O	O
cytokine	NN	O	B-protein
-dependent	NN	O	O
upregulation	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
macrophage	NN	O	I-protein
12-lipoxygenase	NN	O	I-protein
and	NN	O	O
its	NN	O	O
induction	NN	O	O
during	NN	O	O
experimental	NN	O	O
anemia	NN	O	O
suggests	NN	O	O
its	NN	O	O
close	NN	O	O
relatedness	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	B-protein
reticulocyte-type	NN	O	I-protein
15-lipoxygenase	NN	O	I-protein
despite	NN	O	O
their	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
positional	NN	O	O
specificity	NN	O	O
of	NN	O	O
arachidonic	NN	O	O
acid	NN	O	O
oxygenation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interferon-gamma-induced	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
interleukin-4-responsive	NN	O	B-DNA
element	NN	O	I-DNA
of	NN	O	O
CD23b	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
tonsillar	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
role	NN	O	O
in	NN	O	O
transient	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-4-induced	NN	O	B-RNA
CD23b	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
tonsillar	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
distinct	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
with	NN	O	O
different	NN	O	O
mobilities	NN	O	O
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
bind	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
IL-4RE	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
CD23b	NN	O	B-DNA
promoter	NN	O	I-DNA
as	NN	O	O
examined	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
.	NN	O	O

Co-treatment	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IFN-gamma	NN	O	B-protein
induced	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
two	NN	O	O
distinct	NN	O	O
complexes	NN	O	O
,	NN	O	O
a	NN	O	O
new	NN	O	O
complex	NN	O	O
with	NN	O	O
an	NN	O	O
intermediate	NN	O	O
mobility	NN	O	O
.	NN	O	O

The	NN	O	O
IL-4-induced	NN	O	B-protein
complex	NN	O	I-protein
reacted	NN	O	O
with	NN	O	O
anti-STAT	NN	O	B-protein
(	NN	O	I-protein
signal	NN	O	I-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
)	NN	O	I-protein
6	NN	O	I-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
supershift	NN	O	O
whereas	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-gamma-induced	NN	O	B-protein
complex	NN	O	I-protein
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
anti-STAT	NN	O	B-protein
1	NN	O	I-protein
.	NN	O	O

The	NN	O	O
intermediate	NN	O	O
complex	NN	O	O
appeared	NN	O	O
to	NN	O	O
react	NN	O	O
with	NN	O	O
both	NN	O	O
anti-STAT	NN	O	B-protein
6	NN	O	I-protein
and	NN	O	O
anti-STAT	NN	O	B-protein
1	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
IFN-gamma	NN	O	B-protein
alone	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
CD23	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcription	NN	O	O
,	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
a	NN	O	O
transient	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4-induced	NN	O	B-RNA
CD23	NN	O	I-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
IFN-gamma	NN	O	B-protein
within	NN	O	O
2	NN	O	O
h	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
treatment	NN	O	O
in	NN	O	O
these	NN	O	O
tonsillar	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
IL-4RE	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
IL-4-inducible	NN	O	B-DNA
gene	NN	O	I-DNA
can	NN	O	O
accommodate	NN	O	O
both	NN	O	O
IL-4-	NN	O	B-protein
and	NN	O	I-protein
IFN-gamma-activated	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
STAT	NN	O	B-protein
6	NN	O	I-protein
and	NN	O	O
STAT	NN	O	B-protein
1	NN	O	I-protein
,	NN	O	O
either	NN	O	O
in	NN	O	O
homodimeric	NN	O	B-protein
or	NN	O	I-protein
heterodimeric	NN	O	I-protein
forms	NN	O	I-protein
and	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
these	NN	O	O
different	NN	O	O
proteins	NN	O	O
to	NN	O	O
the	NN	O	O
respective	NN	O	O
promoter	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
potential	NN	O	O
regulatory	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
IL-4	NN	O	B-protein
-inducible	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
p56lck	NN	O	B-protein
-independent	NN	O	O
pathway	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
signaling	NN	O	O
involves	NN	O	O
Jun	NN	O	B-protein
kinase	NN	O	I-protein
.	NN	O	O

The	NN	O	O
p56	NN	O	B-protein
Src	NN	O	I-protein
family	NN	O	I-protein
non-receptor	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
critical	NN	O	O
for	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
differentiation	NN	O	O
and	NN	O	O
activation	NN	O	O
.	NN	O	O

Hence	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
p56	NN	O	B-protein
,	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
triggered	NN	O	O
activation	NN	O	O
does	NN	O	O
not	NN	O	O
occur	NN	O	O
.	NN	O	O

We	NN	O	O
now	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
CD2	NN	O	B-protein
-based	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
which	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
p56	NN	O	B-protein
.	NN	O	O

CD2	NN	O	B-protein
-mediated	NN	O	O
interleukin-2	NN	O	B-protein
production	NN	O	O
occurs	NN	O	O
via	NN	O	O
activation	NN	O	O
of	NN	O	O
Jun	NN	O	B-protein
kinase	NN	O	I-protein
in	NN	O	O
cell	NN	O	O
lines	NN	O	O
lacking	NN	O	O
p56	NN	O	B-protein
.	NN	O	O

Jun	NN	O	B-protein
kinase	NN	O	I-protein
then	NN	O	O
facilitates	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
c-Jun/c-Fos	NN	O	B-protein
heterodimers	NN	O	I-protein
to	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
consensus	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
subsequent	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
elucidate	NN	O	O
differences	NN	O	O
between	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
CD2	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
by	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
I	NN	O	I-protein
Tax	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
a	NN	O	O
p16	NN	O	B-cell_line
(	NN	O	I-cell_line
INK4A	NN	O	I-cell_line
)	NN	O	I-cell_line
-negative	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
causative	NN	O	O
agent	NN	O	O
of	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
exact	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
HTLV-I	NN	O	O
contributes	NN	O	O
to	NN	O	O
leukemogenesis	NN	O	O
is	NN	O	O
still	NN	O	O
unclear	NN	O	O
,	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
thought	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

This	NN	O	O
40-kDa	NN	O	O
polypeptide	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
suppressor	NN	O	I-protein
p16	NN	O	B-protein
(	NN	O	I-protein
INK4A	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
Tax	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
that	NN	O	O
in	NN	O	O
turn	NN	O	O
induces	NN	O	O
expression	NN	O	O
of	NN	O	O
factors	NN	O	O
required	NN	O	O
for	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Tax	NN	O	B-protein
can	NN	O	O
also	NN	O	O
activate	NN	O	O
E2F	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
independently	NN	O	O
of	NN	O	O
p16	NN	O	B-protein
(	NN	O	I-protein
INK4A	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
when	NN	O	O
Tax	NN	O	B-protein
is	NN	O	O
coexpressed	NN	O	O
with	NN	O	O
the	NN	O	O
E2F-1	NN	O	B-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
CEM	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
lack	NN	O	O
expression	NN	O	O
of	NN	O	O
p16	NN	O	B-protein
(	NN	O	I-protein
INK4A	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
it	NN	O	O
strongly	NN	O	O
potentiates	NN	O	O
the	NN	O	O
E2F	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	O
construct	NN	O	O
driven	NN	O	O
by	NN	O	O
a	NN	O	O
promoter	NN	O	O
containing	NN	O	O
E2F	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
stimulation	NN	O	O
is	NN	O	O
abrogated	NN	O	O
by	NN	O	O
mutations	NN	O	O
affecting	NN	O	O
the	NN	O	O
E2F-binding	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
Tax	NN	O	B-protein
also	NN	O	O
stimulates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
E2F-1	NN	O	B-DNA
gene	NN	O	I-DNA
itself	NN	O	O
.	NN	O	O

Using	NN	O	O
Tax	NN	O	B-protein
mutants	NN	O	O
that	NN	O	O
fail	NN	O	O
to	NN	O	O
activate	NN	O	O
either	NN	O	O
ATF	NN	O	B-protein
-or	NN	O	O
NF-kappaB-dependent	NN	O	B-DNA
promoters	NN	O	I-DNA
and	NN	O	O
different	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
truncation	NN	O	I-DNA
mutants	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
E2F-1	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
-dependent	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
E2F1	NN	O	B-DNA
gene	NN	O	I-DNA
involves	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
the	NN	O	O
ATF	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
located	NN	O	O
in	NN	O	O
the	NN	O	O
E2F-1	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Inducible	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
:	NN	O	O
an	NN	O	O
autoregulatory	NN	O	O
feedback	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
vascular	NN	O	O
inflammation	NN	O	O
.	NN	O	O

Inducible	NN	O	O
nitric	NN	O	O
oxide	NN	O	O
(	NN	O	O
iNO	NN	O	O
)	NN	O	O
is	NN	O	O
produced	NN	O	O
at	NN	O	O
sites	NN	O	O
of	NN	O	O
vascular	NN	O	O
inflammation	NN	O	O
by	NN	O	O
resident	NN	O	O
and	NN	O	O
nonresident	NN	O	O
vascular	NN	O	B-cell_type
wall	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
its	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
process	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
novel	NN	O	O
function	NN	O	O
of	NN	O	O
iNO	NN	O	O
is	NN	O	O
to	NN	O	O
terminate	NN	O	O
inflammatory	NN	O	O
processes	NN	O	O
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
iNO	NN	O	O
produced	NN	O	O
by	NN	O	O
murine	NN	O	B-cell_line
macrophage-like	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
RAW264.7	NN	O	B-cell_line
,	NN	O	O
can	NN	O	O
inhibit	NN	O	O
cytokine	NN	O	B-protein
-induced	NN	O	O
endothelial	NN	O	O
cell	NN	O	O
activation	NN	O	O
in	NN	O	O
a	NN	O	O
separated	NN	O	O
and	NN	O	O
mixed	NN	O	O
endothelial-RAW264.7	NN	O	B-cell_line
coculture	NN	O	I-cell_line
system	NN	O	I-cell_line
.	NN	O	O

Both	NN	O	O
iNO	NN	O	O
production	NN	O	O
and	NN	O	O
endothelial	NN	O	O
VCAM-1	NN	O	B-protein
expression	NN	O	O
were	NN	O	O
induced	NN	O	O
simultaneously	NN	O	O
with	NN	O	O
bacterial	NN	O	O
LPS	NN	O	O
and	NN	O	O
murine-specific	NN	O	B-protein
IFN-gamma	NN	O	I-protein
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
iNO	NN	O	B-protein
synthase	NN	O	I-protein
(	NN	O	O
iNOS	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
with	NN	O	O
N	NN	O	O
omega-monomethyl-L-arginine	NN	O	O
in	NN	O	O
endothelial-RAW264.7	NN	O	B-cell_line
cocultures	NN	O	I-cell_line
,	NN	O	O
stimulated	NN	O	O
with	NN	O	O
murine-specific	NN	O	B-protein
IFN-gamma	NN	O	I-protein
and	NN	O	O
LPS	NN	O	O
,	NN	O	O
decreased	NN	O	O
iNO	NN	O	O
production	NN	O	O
by	NN	O	O
86	NN	O	O
%	NN	O	O
,	NN	O	O
augmented	NN	O	O
VCAM-1	NN	O	B-protein
and	NN	O	O
iNOS	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
endothelial	NN	O	O
and	NN	O	O
RAW264.7	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
and	NN	O	O
increased	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
the	NN	O	O
endothelial	NN	O	O
cell	NN	O	O
surface	NN	O	O
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
studies	NN	O	O
using	NN	O	O
various	NN	O	O
VCAM-1	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
iNO	NN	O	O
on	NN	O	O
VCAM-1	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
were	NN	O	O
mediated	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
iNO	NN	O	O
on	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
cis-acting	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

Immunofluorescence	NN	O	O
studies	NN	O	O
using	NN	O	O
an	NN	O	O
Ab	NN	O	O
to	NN	O	O
the	NN	O	O
RelA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
subunit	NN	O	I-protein
of	NN	O	O
nuclear	NN	O	O
factor-kappa	NN	O	B-protein
B	NN	O	I-protein
revealed	NN	O	O
that	NN	O	O
iNO	NN	O	O
inhibited	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

These	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
iNO	NN	O	O
attenuates	NN	O	O
iNOS	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
inhibits	NN	O	O
monocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
endogenously	NN	O	O
derived	NN	O	O
iNO	NN	O	O
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
autoregulatory	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
vascular	NN	O	O
inflammation	NN	O	O

-DOCSTART-	O

Detection	NN	O	O
of	NN	O	O
oestrogen	NN	O	B-protein
receptor	NN	O	I-protein
variants	NN	O	I-protein
in	NN	O	O
endometrium	NN	O	O
,	NN	O	O
myometrium	NN	O	O
,	NN	O	O
leiomyoma	NN	O	O
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
comparison	NN	O	O
to	NN	O	O
variants	NN	O	O
present	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

Oestradiol	NN	O	O
has	NN	O	O
mitogenic	NN	O	O
and	NN	O	O
regulatory	NN	O	O
effects	NN	O	O
on	NN	O	O
various	NN	O	O
organs	NN	O	O
and	NN	O	O
cells	NN	O	O
,	NN	O	O
mediated	NN	O	O
mainly	NN	O	O
by	NN	O	O
its	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
aberrant	NN	O	O
ER	NN	O	B-protein
forms	NN	O	O
in	NN	O	O
Oestrogen-dependent	NN	O	O
tumours	NN	O	O
has	NN	O	O
been	NN	O	O
discussed	NN	O	O
in	NN	O	O
correlation	NN	O	O
with	NN	O	O
tumour	NN	O	O
progression	NN	O	O
.	NN	O	O

ER	NN	O	B-protein
variants	NN	O	O
,	NN	O	O
generated	NN	O	O
by	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
human	NN	O	O
breast	NN	O	O
cancer	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
normal	NN	O	O
mammary	NN	O	O
glands	NN	O	O
,	NN	O	O
therefore	NN	O	O
their	NN	O	O
role	NN	O	O
in	NN	O	O
tumorigenesis	NN	O	O
has	NN	O	O
been	NN	O	O
questioned	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
,	NN	O	O
by	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
amplification	NN	O	O
technique	NN	O	O
,	NN	O	O
the	NN	O	O
possible	NN	O	O
existence	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
variants	NN	O	O
in	NN	O	O
other	NN	O	O
normal	NN	O	O
oestrogen	NN	O	O
target	NN	O	O
organs	NN	O	O
and	NN	O	O
cells	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
uterus	NN	O	O
(	NN	O	O
myometrium	NN	O	O
and	NN	O	O
endometrium	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
a	NN	O	O
benign	NN	O	O
uterus	NN	O	O
tumour	NN	O	O
(	NN	O	O
leiomyoma	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
detected	NN	O	O
variant	NN	O	O
ER	NN	O	B-protein
in	NN	O	O
these	NN	O	O
samples	NN	O	O
and	NN	O	O
have	NN	O	O
compared	NN	O	O
the	NN	O	O
variant	NN	O	O
profile	NN	O	O
to	NN	O	O
that	NN	O	O
observed	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

All	NN	O	O
tissues	NN	O	O
and	NN	O	O
cells	NN	O	O
studied	NN	O	O
expressed	NN	O	O
both	NN	O	O
wild-type	NN	O	B-protein
ER	NN	O	I-protein
and	NN	O	O
variant	NN	O	B-protein
species	NN	O	I-protein
.	NN	O	O

Variant	NN	O	O
forms	NN	O	O
encompassed	NN	O	O
ER	NN	O	B-protein
with	NN	O	O
deletions	NN	O	O
of	NN	O	O
exons	NN	O	B-DNA
2	NN	O	I-DNA
,	NN	O	I-DNA
5	NN	O	I-DNA
and	NN	O	I-DNA
7	NN	O	I-DNA
.	NN	O	O

Variants	NN	O	O
with	NN	O	O
exon	NN	O	B-DNA
5	NN	O	I-DNA
deleted	NN	O	O
were	NN	O	O
detected	NN	O	O
only	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

Variants	NN	O	O
with	NN	O	O
exons	NN	O	B-DNA
2	NN	O	I-DNA
and	NN	O	I-DNA
7	NN	O	I-DNA
deleted	NN	O	O
were	NN	O	O
present	NN	O	O
in	NN	O	O
all	NN	O	O
specimens	NN	O	O
tested	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
corroborate	NN	O	O
previous	NN	O	O
findings	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
variants	NN	O	O
is	NN	O	O
not	NN	O	O
a	NN	O	O
characteristic	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

The	NN	O	O
physiological	NN	O	O
significance	NN	O	O
and	NN	O	O
possible	NN	O	O
clinical	NN	O	O
relevance	NN	O	O
of	NN	O	O
the	NN	O	O
variant	NN	O	O
ER	NN	O	B-protein
forms	NN	O	O
remain	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O

-DOCSTART-	O

p21ras	NN	O	B-protein
initiates	NN	O	O
Rac-1	NN	O	B-protein
but	NN	O	O
not	NN	O	O
phosphatidyl	NN	O	B-protein
inositol	NN	O	I-protein
3	NN	O	I-protein
kinase/PKB	NN	O	I-protein
,	NN	O	O
mediated	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

p21ras	NN	O	B-protein
is	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
then	NN	O	O
co-ordinates	NN	O	O
important	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
for	NN	O	O
T	NN	O	B-protein
lymphocyte	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Effector	NN	O	O
pathways	NN	O	O
for	NN	O	O
this	NN	O	O
guanine	NN	O	B-protein
nucleotide	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
serine/threonine	NN	O	B-protein
kinase	NN	O	I-protein
Raf-1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
Ras-related	NN	O	B-protein
GTPase	NN	O	I-protein
Rac-1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
fibroblasts	NN	O	B-cell_type
,	NN	O	O
an	NN	O	O
important	NN	O	O
effector	NN	O	O
for	NN	O	O
the	NN	O	O
Ras	NN	O	B-DNA
oncogene	NN	O	I-DNA
is	NN	O	O
Phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
(	NN	O	O
PtdIns	NN	O	B-protein
3-kinase	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
this	NN	O	O
lipid	NN	O	B-protein
kinase	NN	O	I-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
critical	NN	O	O
Rac-1	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
and	NN	O	O
can	NN	O	O
couple	NN	O	O
p21ras	NN	O	B-protein
to	NN	O	O
cell	NN	O	O
survival	NN	O	O
mechanisms	NN	O	O
via	NN	O	O
the	NN	O	O
serine/threonine	NN	O	B-protein
kinase	NN	O	I-protein
Akt/PKB	NN	O	I-protein
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
PtdIns	NN	O	B-protein
3-kinase	NN	O	I-protein
in	NN	O	O
Ras	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
not	NN	O	O
been	NN	O	O
explored	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
PtdIns	NN	O	B-protein
3-kinase	NN	O	I-protein
to	NN	O	O
initiate	NN	O	O
the	NN	O	O
Rac-1	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
important	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
examined	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
Akt/PKB	NN	O	B-protein
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
Ras	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
Ras	NN	O	B-protein
can	NN	O	O
initiate	NN	O	O
a	NN	O	O
Rac-1	NN	O	B-protein
mediated	NN	O	O
pathway	NN	O	O
that	NN	O	O
regulates	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
function	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
complexes	NN	O	I-protein
.	NN	O	O

PtdIns	NN	O	B-protein
3-kinase	NN	O	I-protein
signals	NN	O	O
can	NN	O	O
not	NN	O	O
mimic	NN	O	O
p21ras	NN	O	B-protein
and	NN	O	O
induce	NN	O	O
the	NN	O	O
Rac	NN	O	B-protein
mediated	NN	O	O
responses	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
neither	NN	O	O
TCR	NN	O	B-protein
or	NN	O	O
Ras	NN	O	B-protein
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
PtdIns	NN	O	B-protein
3-kinase	NN	O	I-protein
.	NN	O	O

PKB	NN	O	B-protein
is	NN	O	O
activated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
triggering	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
;	NN	O	O
PtdIns	NN	O	B-protein
3-kinase	NN	O	I-protein
activity	NN	O	O
is	NN	O	O
both	NN	O	O
required	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
this	NN	O	O
TCR	NN	O	B-protein
response	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
p21ras	NN	O	B-protein
signals	NN	O	O
are	NN	O	O
unable	NN	O	O
to	NN	O	O
induce	NN	O	O
Akt/PKB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
nor	NN	O	O
is	NN	O	O
Ras	NN	O	B-protein
function	NN	O	O
required	NN	O	O
for	NN	O	O
Akt/PKB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
.	NN	O	O

The	NN	O	O
present	NN	O	O
data	NN	O	O
thus	NN	O	O
highlight	NN	O	O
that	NN	O	O
PtdIns	NN	O	B-protein
3-kinase	NN	O	I-protein
and	NN	O	O
Akt/PKB	NN	O	B-protein
are	NN	O	O
not	NN	O	O
universal	NN	O	O
Ras	NN	O	B-protein
effector	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

Ras	NN	O	B-protein
can	NN	O	O
initiate	NN	O	O
Rac-1	NN	O	B-protein
regulated	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
function	NN	O	O
independently	NN	O	O
of	NN	O	O
PtdIns	NN	O	B-protein
3-kinase	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin	NN	O	B-protein
1beta	NN	O	I-protein
mediates	NN	O	O
the	NN	O	O
modulatory	NN	O	O
effects	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
on	NN	O	O
LNCaP	NN	O	B-cell_type
human	NN	O	I-cell_type
prostate	NN	O	I-cell_type
cancer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Proliferative	NN	O	O
and	NN	O	O
secretory	NN	O	O
responses	NN	O	O
in	NN	O	O
androgen-sensitive	NN	O	B-cell_type
prostate	NN	O	I-cell_type
cancer	NN	O	I-cell_type
LNCaP	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
steroid	NN	O	O
and	NN	O	O
peptide	NN	O	O
hormones	NN	O	O
and	NN	O	O
by	NN	O	O
differentiation-promoting	NN	O	O
substances	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
evaluated	NN	O	O
whether	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
that	NN	O	O
exhibit	NN	O	O
anti-tumour	NN	O	O
activity	NN	O	O
in	NN	O	O
haematopoietic	NN	O	O
and	NN	O	O
solid	NN	O	O
tumours	NN	O	O
influence	NN	O	O
growth	NN	O	O
and	NN	O	O
secretion	NN	O	O
in	NN	O	O
the	NN	O	O
LNCaP	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

For	NN	O	O
this	NN	O	O
purpose	NN	O	O
,	NN	O	O
LNCaP	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
monocyte-conditioned	NN	O	O
medium	NN	O	O
(	NN	O	O
MCM	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
proliferation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
expression	NN	O	O
of	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
AR	NN	O	B-protein
)	NN	O	O
and	NN	O	O
secretion	NN	O	O
of	NN	O	O
prostate-specific	NN	O	B-protein
antigen	NN	O	I-protein
(	NN	O	O
PSA	NN	O	B-protein
)	NN	O	O
were	NN	O	O
assessed	NN	O	O
.	NN	O	O

Conditioned	NN	O	O
medium	NN	O	O
from	NN	O	O
monocytes	NN	O	B-cell_type
reduced	NN	O	O
proliferation	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Incubation	NN	O	O
with	NN	O	O
40	NN	O	O
%	NN	O	O
MCM	NN	O	O
caused	NN	O	O
a	NN	O	O
50	NN	O	O
%	NN	O	O
reduction	NN	O	O
in	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

AR	NN	O	B-protein
protein	NN	O	I-protein
decreased	NN	O	O
by	NN	O	O
70	NN	O	O
%	NN	O	O
and	NN	O	O
PSA	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
supernatants	NN	O	O
from	NN	O	O
LNCaP	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
reduced	NN	O	O
by	NN	O	O
approximately	NN	O	O
80	NN	O	O
%	NN	O	O
following	NN	O	O
treatment	NN	O	O
with	NN	O	O
MCM	NN	O	O
.	NN	O	O

We	NN	O	O
focused	NN	O	O
on	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
two	NN	O	O
major	NN	O	O
products	NN	O	O
of	NN	O	O
activated	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
prostaglandin	NN	O	O
E2	NN	O	O
and	NN	O	O
interleukin	NN	O	B-protein
1beta	NN	O	I-protein
(	NN	O	O
IL-1beta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
to	NN	O	O
the	NN	O	O
MCM	NN	O	O
modulatory	NN	O	O
action	NN	O	O
.	NN	O	O

LNCaP	NN	O	B-cell_line
cells	NN	O	I-cell_line
treated	NN	O	O
with	NN	O	O
prostaglandin	NN	O	O
E2	NN	O	O
showed	NN	O	O
neither	NN	O	O
a	NN	O	O
reduction	NN	O	O
in	NN	O	O
proliferation	NN	O	O
nor	NN	O	O
a	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
AR	NN	O	B-protein
and	NN	O	O
PSA	NN	O	B-protein
levels	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
MCM	NN	O	O
on	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
,	NN	O	O
AR	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
PSA	NN	O	B-protein
secretion	NN	O	O
were	NN	O	O
abolished	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
MCM	NN	O	O
with	NN	O	O
a	NN	O	O
neutralizing	NN	O	O
anti-IL-1beta	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
recombinant	NN	O	O
IL-1beta	NN	O	B-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
replace	NN	O	O
MCM	NN	O	O
for	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
proliferation	NN	O	O
and	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
AR	NN	O	B-protein
and	NN	O	O
PSA	NN	O	B-protein
proteins	NN	O	O
.	NN	O	O

LNCaP	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
shown	NN	O	O
to	NN	O	O
express	NN	O	O
the	NN	O	O
IL-1beta	NN	O	B-protein
receptor	NN	O	O
type	NN	O	O
1	NN	O	O
,	NN	O	O
which	NN	O	O
transduces	NN	O	O
IL-1beta	NN	O	B-protein
signal	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
reveal	NN	O	O
that	NN	O	O
monocyte-derived	NN	O	B-protein
IL-1beta	NN	O	I-protein
inhibits	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
androgen-responsive	NN	O	B-cell_type
prostate	NN	O	I-cell_type
tumour	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
reduces	NN	O	O
AR	NN	O	B-protein
and	NN	O	O
PSA	NN	O	B-protein
levels	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
and	NN	O	I-DNA
IL-4	NN	O	I-DNA
genes	NN	O	I-DNA
to	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-3	NN	O	I-protein
.	NN	O	O

The	NN	O	O
cytokines	NN	O	B-protein
IL-4	NN	O	B-protein
and	NN	O	O
IL-5	NN	O	B-protein
are	NN	O	O
often	NN	O	O
coordinately	NN	O	O
produced	NN	O	O
by	NN	O	O
Th2	NN	O	B-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
in	NN	O	O
asthma	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
unclear	NN	O	O
whether	NN	O	O
similar	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
underlie	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
genes	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-3	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
Th2	NN	O	B-cell_line
but	NN	O	I-cell_line
not	NN	O	I-cell_line
Th1	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
different	NN	O	O
stimuli	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
different	NN	O	O
study	NN	O	O
,	NN	O	O
GATA-3	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
sufficient	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
other	NN	O	O
Th2	NN	O	B-DNA
cytokine	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
drive	NN	O	O
IL-5	NN	O	B-protein
but	NN	O	O
not	NN	O	O
IL-4	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Also	NN	O	O
,	NN	O	O
in	NN	O	O
Th2	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
antisense	NN	O	O
GATA-3	NN	O	B-protein
RNA	NN	O	O
inhibits	NN	O	O
IL-5	NN	O	B-protein
but	NN	O	O
not	NN	O	O
IL-4	NN	O	B-protein
promoter	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-5	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
GATA-3	NN	O	B-protein
involves	NN	O	O
high	NN	O	O
affinity	NN	O	O
binding	NN	O	O
of	NN	O	O
GATA-3	NN	O	B-protein
to	NN	O	O
an	NN	O	O
inverted	NN	O	O
GATA	NN	O	B-DNA
repeat	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-5	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
chain/CD25	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
interleukin	NN	O	I-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-2	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain/CD25	NN	O	I-protein
is	NN	O	O
strongly	NN	O	O
induced	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
after	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cell	NN	O	I-protein
(	NN	O	I-protein
NF-AT	NN	O	I-protein
)	NN	O	I-protein
factors	NN	O	I-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
CD25	NN	O	B-DNA
promoter	NN	O	I-DNA
induction	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NF-ATp	NN	O	B-protein
and	NN	O	O
NF-ATc	NN	O	B-protein
bind	NN	O	O
to	NN	O	O
two	NN	O	O
sites	NN	O	O
around	NN	O	O
positions	NN	O	B-DNA
-585	NN	O	I-DNA
and	NN	O	I-DNA
-650	NN	O	I-DNA
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
CD25	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Immediately	NN	O	O
3	NN	O	O
'	NN	O	O
from	NN	O	O
these	NN	O	O
NF-AT	NN	O	B-DNA
motifs	NN	O	I-DNA
,	NN	O	O
nonconsensus	NN	O	B-DNA
sites	NN	O	I-DNA
are	NN	O	O
located	NN	O	O
for	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
AP-1-like	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Mutations	NN	O	O
of	NN	O	O
sites	NN	O	O
that	NN	O	O
suppress	NN	O	O
NF-AT	NN	O	B-protein
binding	NN	O	O
impair	NN	O	O
the	NN	O	O
induction	NN	O	O
and	NN	O	O
strong	NN	O	O
NF-ATp	NN	O	B-protein
-mediated	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
CD25	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
NF-ATp	NN	O	B-protein
-deficient	NN	O	O
mice	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CD25	NN	O	B-protein
is	NN	O	O
severely	NN	O	O
impaired	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
a	NN	O	O
delayed	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
after	NN	O	O
T	NN	O	O
cell	NN	O	O
receptor	NN	O	O
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
/	NN	O	O
CD3	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
indicate	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
for	NN	O	O
NF-AT	NN	O	B-protein
in	NN	O	O
the	NN	O	O
faithful	NN	O	O
expression	NN	O	O
of	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
IL-2	NN	O	I-protein
receptors	NN	O	I-protein
and	NN	O	O
a	NN	O	O
close	NN	O	O
link	NN	O	O
between	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
chain	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
NF-AT	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
coding	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
FY*B	NN	O	B-DNA
allele	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
Duffy	NN	O	B-DNA
chemokine	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
altered	NN	O	O
erythrocyte	NN	O	O
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
Duffy	NN	O	O
blood	NN	O	O
group	NN	O	O
system	NN	O	O
is	NN	O	O
of	NN	O	O
clinical	NN	O	O
and	NN	O	O
biological	NN	O	O
significance	NN	O	O
.	NN	O	O

Antibodies	NN	O	B-protein
to	NN	O	O
Duffy	NN	O	B-protein
antigens	NN	O	I-protein
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
some	NN	O	O
cases	NN	O	O
of	NN	O	O
transfusion	NN	O	O
incompatibility	NN	O	O
and	NN	O	O
newborn	NN	O	O
hemolytic	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
Duffy	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
a	NN	O	O
receptor	NN	O	O
for	NN	O	O
the	NN	O	O
Plasmodium	NN	O	B-protein
vivax	NN	O	I-protein
erythrocyte-binding	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
is	NN	O	O
also	NN	O	O
a	NN	O	O
receptor	NN	O	O
for	NN	O	O
various	NN	O	O
chemokines	NN	O	B-protein
(	NN	O	O
thus	NN	O	O
renamed	NN	O	O
Duffy	NN	O	B-protein
Antigen	NN	O	I-protein
Receptor	NN	O	I-protein
for	NN	O	I-protein
Chemokines	NN	O	I-protein
[	NN	O	O
DARC	NN	O	B-protein
]	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
Duffy	NN	O	B-protein
polymorphic	NN	O	I-protein
antigens	NN	O	I-protein
,	NN	O	O
Fya	NN	O	B-protein
and	NN	O	O
Fyb	NN	O	B-protein
(	NN	O	O
coded	NN	O	O
by	NN	O	O
the	NN	O	O
FY*A	NN	O	B-DNA
and	NN	O	I-DNA
FY*B	NN	O	I-DNA
alleles	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
are	NN	O	O
present	NN	O	O
on	NN	O	O
erythrocyte	NN	O	B-protein
membranes	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Fy	NN	O	O
(	NN	O	O
a-b-	NN	O	O
)	NN	O	O
phenotype	NN	O	O
is	NN	O	O
the	NN	O	O
predominant	NN	O	O
one	NN	O	O
in	NN	O	O
populations	NN	O	O
of	NN	O	O
black	NN	O	O
people	NN	O	O
and	NN	O	O
also	NN	O	O
occurs	NN	O	O
in	NN	O	O
other	NN	O	O
populations	NN	O	O
,	NN	O	O
including	NN	O	O
some	NN	O	O
non-Ashkenazi	NN	O	O
Jewish	NN	O	O
groups	NN	O	O
.	NN	O	O

The	NN	O	O
Fy	NN	O	O
(	NN	O	O
a-b-	NN	O	O
)	NN	O	O
phenotype	NN	O	O
has	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
FY*B	NN	O	B-DNA
promoter	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
GATA	NN	O	B-DNA
box	NN	O	I-DNA
that	NN	O	O
abolishes	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
erythrocyte	NN	O	B-protein
Duffy	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
describe	NN	O	O
here	NN	O	O
a	NN	O	O
novel	NN	O	O
mutation	NN	O	O
,	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
FY*B	NN	O	B-DNA
coding	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
271C	NN	O	O
--	NN	O	O
>	NN	O	O
T	NN	O	O
)	NN	O	O
,	NN	O	O
that	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
some	NN	O	O
Fy	NN	O	O
(	NN	O	O
b-	NN	O	O
)	NN	O	O
phenotypes	NN	O	O
among	NN	O	O
non-Ashkenazi	NN	O	O
Jews	NN	O	O
and	NN	O	O
among	NN	O	O
Brazilian	NN	O	O
blacks	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
Fy	NN	O	O
(	NN	O	O
b-	NN	O	O
)	NN	O	O
individuals	NN	O	O
,	NN	O	O
who	NN	O	O
have	NN	O	O
wild-type	NN	O	B-DNA
FY*B	NN	O	I-DNA
GATA	NN	O	I-DNA
and	NN	O	O
carry	NN	O	O
the	NN	O	O
previously	NN	O	O
described	NN	O	O
304G	NN	O	O
--	NN	O	O
>	NN	O	O
A	NN	O	O
substitution	NN	O	O
.	NN	O	O

The	NN	O	O
271C	NN	O	O
--	NN	O	O
>	NN	O	O
T	NN	O	O
and	NN	O	O
304G	NN	O	O
--	NN	O	O
>	NN	O	O
A	NN	O	O
can	NN	O	O
be	NN	O	O
identified	NN	O	O
by	NN	O	O
restriction	NN	O	B-DNA
enzyme-generated	NN	O	I-DNA
restriction	NN	O	I-DNA
fragment	NN	O	I-DNA
length	NN	O	I-DNA
polymorphisms	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
271C	NN	O	O
--	NN	O	O
>	NN	O	O
T	NN	O	O
substitution	NN	O	O
represents	NN	O	O
a	NN	O	O
considerable	NN	O	O
change	NN	O	O
in	NN	O	O
chemical	NN	O	O
nature	NN	O	O
(	NN	O	O
Arg91	NN	O	O
--	NN	O	O
>	NN	O	O
Cys	NN	O	O
)	NN	O	O
,	NN	O	O
one	NN	O	O
which	NN	O	O
may	NN	O	O
affect	NN	O	O
the	NN	O	O
antigenic	NN	O	O
determinants	NN	O	O
of	NN	O	O
DARC	NN	O	B-protein
,	NN	O	O
and	NN	O	O
thus	NN	O	O
be	NN	O	O
of	NN	O	O
clinical	NN	O	O
significance	NN	O	O
.	NN	O	O

The	NN	O	O
mutation	NN	O	O
may	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
some	NN	O	O
physiological	NN	O	O
roles	NN	O	O
of	NN	O	O
DARC	NN	O	B-protein
and	NN	O	O
be	NN	O	O
of	NN	O	O
interest	NN	O	O
in	NN	O	O
malaria	NN	O	O
research	NN	O	O
and	NN	O	O
in	NN	O	O
studies	NN	O	O
of	NN	O	O
population	NN	O	O
genetics	NN	O	O
.	NN	O	O

-DOCSTART-	O

Establishment	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
EBV-positive	NN	O	B-cell_line
and	NN	O	I-cell_line
EBV-negative	NN	O	I-cell_line
primary	NN	O	I-cell_line
effusion	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
harbouring	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
type-8	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
report	NN	O	O
on	NN	O	O
the	NN	O	O
establishment	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
two	NN	O	O
novel	NN	O	O
lymphoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
CRO-AP/3	NN	O	B-cell_line
and	NN	O	O
CRO-AP/5	NN	O	B-cell_line
)	NN	O	O
which	NN	O	O
carry	NN	O	O
infection	NN	O	O
by	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
type-8	NN	O	O
(	NN	O	O
HHV-8	NN	O	O
)	NN	O	O
and	NN	O	O
have	NN	O	O
derived	NN	O	O
from	NN	O	O
AIDS-related	NN	O	O
primary	NN	O	O
effusion	NN	O	O
lymphoma	NN	O	O
(	NN	O	O
PEL	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

These	NN	O	O
two	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
are	NN	O	O
representative	NN	O	O
of	NN	O	O
different	NN	O	O
virologic	NN	O	O
subtypes	NN	O	O
of	NN	O	O
PEL	NN	O	B-cell_line
,	NN	O	O
i.e	NN	O	O
.	NN	O	O
HHV-8+/EBV-	NN	O	B-cell_line
PEL	NN	O	I-cell_line
in	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
CRO-AP/3	NN	O	B-cell_line
and	NN	O	I-cell_line
HHV-8+/EBV+	NN	O	I-cell_line
PEL	NN	O	I-cell_line
in	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
CRO-AP/5	NN	O	B-cell_line
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
PEL	NN	O	B-cell_line
,	NN	O	O
both	NN	O	O
CRO-AP/3	NN	O	B-cell_line
and	NN	O	O
CRO-AP/5	NN	O	B-cell_line
expressed	NN	O	O
indeterminate	NN	O	O
(	NN	O	O
i.e.	NN	O	O
non-B	NN	O	O
,	NN	O	O
non-T	NN	O	O
)	NN	O	O
phenotypes	NN	O	O
although	NN	O	O
immunogenotypic	NN	O	O
studies	NN	O	O
documented	NN	O	O
their	NN	O	O
B-cell	NN	O	B-cell_type
origin	NN	O	I-cell_type
.	NN	O	O

Both	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
are	NN	O	O
devoid	NN	O	O
of	NN	O	O
genetic	NN	O	O
lesions	NN	O	O
of	NN	O	O
c-MYC	NN	O	B-protein
,	NN	O	O
BCL-2	NN	O	B-protein
and	NN	O	O
p53	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
gross	NN	O	O
rearrangements	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
.	NN	O	O

Detailed	NN	O	O
histogenetic	NN	O	O
characterization	NN	O	O
of	NN	O	O
these	NN	O	O
novel	NN	O	O
PEL	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
suggests	NN	O	O
that	NN	O	O
PEL	NN	O	B-cell_line
may	NN	O	O
derive	NN	O	O
from	NN	O	O
a	NN	O	O
post-germinal	NN	O	B-cell_line
centre	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
which	NN	O	O
has	NN	O	O
undergone	NN	O	O
pre-terminal	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
CRO-AP/3	NN	O	B-cell_line
and	NN	O	O
CRO-AP/5	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
may	NN	O	O
provide	NN	O	O
a	NN	O	O
valuable	NN	O	O
model	NN	O	O
for	NN	O	O
clarifying	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
PEL	NN	O	B-cell_line
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
these	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
may	NN	O	O
help	NN	O	O
understand	NN	O	O
the	NN	O	O
relative	NN	O	O
contribution	NN	O	O
of	NN	O	O
HHV-8	NN	O	O
and	NN	O	O
EBV	NN	O	O
to	NN	O	O
PEL	NN	O	O
growth	NN	O	O
and	NN	O	O
development	NN	O	O
and	NN	O	O
may	NN	O	O
facilitate	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
recurrent	NN	O	O
cytogenetic	NN	O	O
abnormalities	NN	O	O
highlighting	NN	O	O
putative	NN	O	O
novel	NN	O	O
cancer	NN	O	O
related	NN	O	O
loci	NN	O	O
relevant	NN	O	O
to	NN	O	O
PEL	NN	O	B-cell_line
.	NN	O	O

-DOCSTART-	O

Two-site	NN	O	O
interaction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
with	NN	O	O
activated	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
belonging	NN	O	O
to	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	I-protein
NFAT	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
regulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
other	NN	O	O
inducible	NN	O	O
genes	NN	O	O
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
functions	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
directly	NN	O	O
controlled	NN	O	O
by	NN	O	O
the	NN	O	O
calcium-	NN	O	B-protein
and	NN	O	I-protein
calmodulin-	NN	O	I-protein
dependent	NN	O	I-protein
phosphatase	NN	O	I-protein
calcineurin	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
to	NN	O	O
NFAT	NN	O	B-protein
is	NN	O	O
substantially	NN	O	O
increased	NN	O	O
when	NN	O	O
calcineurin	NN	O	B-protein
is	NN	O	O
activated	NN	O	O
with	NN	O	O
calmodulin	NN	O	B-protein
and	NN	O	O
calcium	NN	O	O
.	NN	O	O

FK506.FKBP12	NN	O	O
drug-immunophilin	NN	O	O
complexes	NN	O	O
inhibited	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
with	NN	O	O
activated	NN	O	O
calcineurin	NN	O	B-protein
much	NN	O	O
more	NN	O	O
effectively	NN	O	O
than	NN	O	O
they	NN	O	O
inhibited	NN	O	O
the	NN	O	O
interaction	NN	O	O
with	NN	O	O
inactive	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
interaction	NN	O	O
with	NN	O	O
activated	NN	O	O
calcineurin	NN	O	B-protein
involved	NN	O	O
the	NN	O	O
enzyme	NN	O	B-DNA
active	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
NFAT	NN	O	B-protein
is	NN	O	O
targeted	NN	O	O
to	NN	O	O
inactive	NN	O	O
calcineurin	NN	O	B-protein
at	NN	O	O
a	NN	O	O
region	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-DNA
active	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
Aramburu	NN	O	O
,	NN	O	O
J.	NN	O	O
,	NN	O	O
Garcia-Cozar	NN	O	O
,	NN	O	O
F.	NN	O	O
J.	NN	O	O
,	NN	O	O
Raghavan	NN	O	O
,	NN	O	O
A.	NN	O	O
,	NN	O	O
Okamura	NN	O	O
,	NN	O	O
H.	NN	O	O
,	NN	O	O
Rao	NN	O	O
,	NN	O	O
A.	NN	O	O
,	NN	O	O
and	NN	O	O
Hogan	NN	O	O
,	NN	O	O
P.	NN	O	O
G.	NN	O	O
(	NN	O	O
1998	NN	O	O
)	NN	O	O
Mol.	NN	O	O
Cell	NN	O	O
1	NN	O	O
,	NN	O	O
627-637	NN	O	O
)	NN	O	O
;	NN	O	O
this	NN	O	O
region	NN	O	O
is	NN	O	O
also	NN	O	O
involved	NN	O	O
in	NN	O	O
NFAT	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
activated	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
since	NN	O	O
binding	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
an	NN	O	O
NFAT	NN	O	O
peptide	NN	O	O
spanning	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
docking	NN	O	O
site	NN	O	O
on	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

The	NN	O	O
interacting	NN	O	O
surfaces	NN	O	O
are	NN	O	O
located	NN	O	O
on	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
A	NN	O	I-protein
chain	NN	O	I-protein
and	NN	O	O
on	NN	O	O
an	NN	O	O
86-amino	NN	O	B-protein
acid	NN	O	I-protein
fragment	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
regulatory	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

NFAT	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
catalytic	NN	O	I-protein
domain	NN	O	I-protein
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
autoinhibitory	NN	O	I-protein
domain	NN	O	I-protein
and	NN	O	O
the	NN	O	O
RII	NN	O	O
substrate	NN	O	O
peptide	NN	O	O
,	NN	O	O
which	NN	O	O
bind	NN	O	O
in	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
active	NN	O	I-protein
site	NN	O	I-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
docking	NN	O	O
site	NN	O	O
peptide	NN	O	O
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
active	NN	O	B-protein
site	NN	O	I-protein
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
NFAT	NN	O	B-protein
is	NN	O	O
targeted	NN	O	O
to	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
catalytic	NN	O	I-protein
domain	NN	O	I-protein
that	NN	O	O
does	NN	O	O
not	NN	O	O
overlap	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
active	NN	O	I-protein
site	NN	O	I-protein
.	NN	O	O

Upon	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
displacement	NN	O	O
of	NN	O	O
the	NN	O	O
autoinhibitory	NN	O	B-protein
domain	NN	O	I-protein
by	NN	O	O
calmodulin	NN	O	B-protein
binding	NN	O	O
allows	NN	O	O
NFAT	NN	O	B-protein
to	NN	O	O
bind	NN	O	O
additionally	NN	O	O
to	NN	O	O
the	NN	O	O
calcineurin	NN	O	B-protein
active	NN	O	I-protein
site	NN	O	I-protein
,	NN	O	O
thus	NN	O	O
positioning	NN	O	O
NFAT	NN	O	B-protein
for	NN	O	O
immediate	NN	O	O
dephosphorylation	NN	O	O
at	NN	O	O
functional	NN	O	O
phosphoserine	NN	O	O
residues	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
induction	NN	O	O
in	NN	O	O
CD45RO+	NN	O	B-cell_type
and	NN	O	I-cell_type
CD45RA+	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
subsets	NN	O	I-cell_type
during	NN	O	O
aging	NN	O	O
.	NN	O	O

An	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
memory	NN	O	B-cell_type
to	NN	O	I-cell_type
naive	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
postulated	NN	O	O
to	NN	O	O
underlie	NN	O	O
immune	NN	O	O
hyporesponsiveness	NN	O	O
accompanying	NN	O	O
aging	NN	O	O
.	NN	O	O

Our	NN	O	O
analyses	NN	O	O
of	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NFkappaB	NN	O	B-protein
)	NN	O	O
in	NN	O	O
activated	NN	O	O
memory	NN	O	B-cell_type
(	NN	O	I-cell_type
CD45RO+	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	I-cell_type
naive	NN	O	I-cell_type
(	NN	O	I-cell_type
CD45RA+	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
subsets	NN	O	I-cell_type
from	NN	O	O
young	NN	O	O
and	NN	O	O
elderly	NN	O	O
donors	NN	O	O
has	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
,	NN	O	O
regardless	NN	O	O
of	NN	O	O
donor	NN	O	O
age	NN	O	O
,	NN	O	O
memory	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
not	NN	O	O
significantly	NN	O	O
altered	NN	O	O
in	NN	O	O
their	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
TNF-alpha	NN	O	B-protein
-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
.	NN	O	O

Although	NN	O	O
treatment	NN	O	O
with	NN	O	O
TNF-alpha	NN	O	B-protein
induced	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
in	NN	O	O
both	NN	O	O
memory	NN	O	B-cell_type
and	NN	O	I-cell_type
naive	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
subsets	NN	O	I-cell_type
,	NN	O	O
irrespective	NN	O	O
of	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
the	NN	O	O
donor	NN	O	O
,	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
induced	NN	O	O
NFkappaB	NN	O	B-protein
were	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
both	NN	O	O
subsets	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
the	NN	O	O
elderly	NN	O	O
,	NN	O	O
when	NN	O	O
compared	NN	O	O
to	NN	O	O
those	NN	O	O
in	NN	O	O
young	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
regulation	NN	O	O
revealed	NN	O	O
that	NN	O	O
TNF-alpha	NN	O	B-protein
-mediated	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
in	NN	O	O
both	NN	O	O
memory	NN	O	B-cell_type
and	NN	O	I-cell_type
naive	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
elderly	NN	O	O
was	NN	O	O
severely	NN	O	O
impaired	NN	O	O
,	NN	O	O
thus	NN	O	O
contributing	NN	O	O
to	NN	O	O
the	NN	O	O
lowered	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
observed	NN	O	O
NFkappaB	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
this	NN	O	O
age-related	NN	O	O
decrease	NN	O	O
in	NN	O	O
induction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
NFkappaB	NN	O	I-protein
correlated	NN	O	O
with	NN	O	O
decrease	NN	O	O
in	NN	O	O
intracellular	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	O
expression	NN	O	O
and	NN	O	O
anti-CD3	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
of	NN	O	O
both	NN	O	O
memory	NN	O	B-cell_type
and	NN	O	I-cell_type
naive	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
subsets	NN	O	I-cell_type
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
age-related	NN	O	O
hyporesponsiveness	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
attributed	NN	O	O
to	NN	O	O
a	NN	O	O
skewing	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
population	NN	O	O
towards	NN	O	O
a	NN	O	O
memory	NN	O	O
phenotype	NN	O	O
in	NN	O	O
the	NN	O	O
elderly	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cyclosporin	NN	O	O
A-resistant	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
requires	NN	O	O
activity	NN	O	O
of	NN	O	O
okadaic	NN	O	B-protein
acid-sensitive	NN	O	I-protein
serine/threonine	NN	O	I-protein
phosphatases	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
requires	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
costimulation	NN	O	O
through	NN	O	O
accessory	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
recently	NN	O	O
that	NN	O	O
okadaic	NN	O	B-protein
acid-sensitive	NN	O	I-protein
Ser/Thr	NN	O	I-protein
phosphatases	NN	O	I-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
a	NN	O	O
cyclosporin	NN	O	O
A-insensitive	NN	O	O
pathway	NN	O	O
that	NN	O	O
selectively	NN	O	O
transmits	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
whether	NN	O	O
activities	NN	O	O
of	NN	O	O
these	NN	O	O
phosphatases	NN	O	O
are	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
both	NN	O	O
activated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
activated	NN	O	B-cell_line
tumorigenic	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
blocked	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
active	NN	O	O
at	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
were	NN	O	O
differentially	NN	O	O
influenced	NN	O	O
:	NN	O	O
upon	NN	O	O
down-modulation	NN	O	O
of	NN	O	O
okadaic	NN	O	B-protein
acid-sensitive	NN	O	I-protein
phosphatases	NN	O	I-protein
,	NN	O	O
transactivation	NN	O	O
by	NN	O	O
octamer	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
NF	NN	O	B-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
proteins	NN	O	I-protein
was	NN	O	O
abrogated	NN	O	O
,	NN	O	O
while	NN	O	O
transactivation	NN	O	O
by	NN	O	O
AP-1	NN	O	B-protein
proteins	NN	O	I-protein
was	NN	O	I-protein
even	NN	O	I-protein
enhanced	NN	O	I-protein
.	NN	O	I-protein

-DOCSTART-	O

YM268	NN	O	O
increases	NN	O	O
the	NN	O	O
glucose	NN	O	O
uptake	NN	O	O
,	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
glucose	NN	O	B-protein
transporter	NN	O	I-protein
in	NN	O	O
3T3-L1	NN	O	B-cell_type
adipocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
purpose	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
examine	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
bis	NN	O	O
[	NN	O	O
4-	NN	O	O
[	NN	O	O
2	NN	O	O
,	NN	O	O
4-dioxo-5-thiazolidinyl	NN	O	O
)	NN	O	O
methyl	NN	O	O
]	NN	O	O
phenyl	NN	O	O
]	NN	O	O
methane	NN	O	O
(	NN	O	O
YM-268	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
thiazolidinedione	NN	O	O
derivative	NN	O	O
,	NN	O	O
on	NN	O	O
glucose	NN	O	O
uptake	NN	O	O
,	NN	O	O
adipocyte	NN	O	B-cell_type
differentiation	NN	O	O
through	NN	O	O
peroxisome	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
gamma	NN	O	I-protein
(	NN	O	O
PPARgamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
(	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
)	NN	O	O
activity	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

YM268	NN	O	O
and	NN	O	O
pioglitazone	NN	O	O
dose-dependently	NN	O	O
increased	NN	O	O
the	NN	O	O
2-deoxyglucose	NN	O	O
uptake	NN	O	O
in	NN	O	O
3T3-L1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

YM268	NN	O	O
facilitated	NN	O	O
the	NN	O	O
insulin-stimulated	NN	O	O
triglyceride	NN	O	O
accumulation	NN	O	O
in	NN	O	O
3T3-L1	NN	O	B-cell_type
adipocytes	NN	O	I-cell_type
and	NN	O	O
increased	NN	O	O
the	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
fatty	NN	O	B-protein
acid-binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

YM268	NN	O	O
,	NN	O	O
with	NN	O	O
and	NN	O	O
without	NN	O	O
insulin	NN	O	O
,	NN	O	O
increased	NN	O	O
the	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
glucose	NN	O	B-protein
transporter	NN	O	I-protein
isoforms	NN	O	O
such	NN	O	O
as	NN	O	O
GLUT1	NN	O	B-protein
and	NN	O	O
GLUT4	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
enhancement	NN	O	O
of	NN	O	O
adipocyte	NN	O	B-cell_type
differentiation	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
YM268	NN	O	O
and	NN	O	O
pioglitazone	NN	O	O
showed	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
PPARgamma	NN	O	B-protein
ligand	NN	O	I-protein
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
responsible	NN	O	O
for	NN	O	O
adipogenesis	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
possible	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
increased	NN	O	O
activity	NN	O	O
of	NN	O	O
PI	NN	O	B-protein
3-kinase	NN	O	I-protein
in	NN	O	O
YM268-stimulated	NN	O	O
glucose	NN	O	O
uptake	NN	O	O
,	NN	O	O
the	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
was	NN	O	O
estimated	NN	O	O
by	NN	O	O
measuring	NN	O	O
the	NN	O	O
phosphatidylinositol-3	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
5-trisphosphate	NN	O	O
(	NN	O	O
PI-3	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
5-P3	NN	O	O
)	NN	O	O
concentration	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Insulin	NN	O	O
dose-dependently	NN	O	O
increased	NN	O	O
the	NN	O	O
PI-3	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
5-P3	NN	O	O
production	NN	O	O
but	NN	O	O
YM268	NN	O	O
had	NN	O	O
no	NN	O	O
significant	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
insulin-dependent	NN	O	O
and	NN	O	O
-independent	NN	O	O
PI	NN	O	O
3-kinase	NN	O	O
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
YM268	NN	O	O
increased	NN	O	O
glucose	NN	O	O
uptake	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
accounted	NN	O	O
for	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
the	NN	O	O
enhancement	NN	O	O
of	NN	O	O
GLUT1	NN	O	B-protein
and	NN	O	O
GLUT4	NN	O	B-protein
expression	NN	O	O
through	NN	O	O
PPARgamma	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
PPARgamma	NN	O	B-protein
-	NN	O	O
bigger	NN	O	O
than	NN	O	O
fat	NN	O	O
.	NN	O	O

Work	NN	O	O
reported	NN	O	O
over	NN	O	O
the	NN	O	O
past	NN	O	O
year	NN	O	O
has	NN	O	O
provided	NN	O	O
insights	NN	O	O
into	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
whereby	NN	O	O
ligand	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
peroxisome	NN	O	I-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
gamma	NN	O	I-protein
(	NN	O	O
PPARgamma	NN	O	B-protein
)	NN	O	O
regulates	NN	O	O
systemic	NN	O	O
glucose	NN	O	O
and	NN	O	O
lipid	NN	O	O
homeostasis	NN	O	O
.	NN	O	O

PPARgamma	NN	O	B-protein
has	NN	O	O
also	NN	O	O
been	NN	O	O
implicated	NN	O	O
recently	NN	O	O
in	NN	O	O
the	NN	O	O
biology	NN	O	O
of	NN	O	O
monocytes	NN	O	O
and	NN	O	O
in	NN	O	O
cell-cycle	NN	O	O
regulation	NN	O	O
and	NN	O	O
cancer	NN	O	O
.	NN	O	O

Polyunsaturated	NN	O	O
fatty	NN	O	O
acids	NN	O	O
and	NN	O	O
eicosanoids	NN	O	O
bind	NN	O	O
and	NN	O	O
activate	NN	O	O
PPARgamma	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
these	NN	O	O
lipids	NN	O	O
may	NN	O	O
serve	NN	O	O
as	NN	O	O
hormonal	NN	O	O
regulators	NN	O	O
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
biological	NN	O	O
processes	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
caspases	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
and	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

Apoptosis	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
removal	NN	O	O
of	NN	O	O
potentially	NN	O	O
autoreactive	NN	O	O
or	NN	O	O
useless	NN	O	O
T	NN	O	O
cells	NN	O	O
during	NN	O	O
thymic	NN	O	O
selection	NN	O	O
and	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
periphery	NN	O	O
.	NN	O	O

Specific	NN	O	O
families	NN	O	O
of	NN	O	O
receptors	NN	O	B-protein
,	NN	O	O
kinases	NN	O	B-protein
,	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
cysteine	NN	O	B-protein
proteases	NN	O	I-protein
,	NN	O	O
termed	NN	O	B-protein
caspases	NN	O	I-protein
,	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
cascade	NN	O	O
leading	NN	O	O
to	NN	O	O
proteolysis	NN	O	O
of	NN	O	O
specific	NN	O	O
substrates	NN	O	O
and	NN	O	O
to	NN	O	O
morphological	NN	O	O
changes	NN	O	O
associated	NN	O	O
with	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Although	NN	O	O
common	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
cascade	NN	O	O
are	NN	O	O
shared	NN	O	O
between	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
it	NN	O	O
appears	NN	O	O
that	NN	O	O
cell-specific	NN	O	B-protein
factors	NN	O	I-protein
can	NN	O	O
influence	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
given	NN	O	O
apoptotic	NN	O	O
stimuli	NN	O	O
.	NN	O	O

Characterization	NN	O	O
and	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
basic	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
different	NN	O	O
pathways	NN	O	O
protecting	NN	O	O
or	NN	O	O
leading	NN	O	O
to	NN	O	O
cell	NN	O	O
death	NN	O	O
may	NN	O	O
provide	NN	O	O
novel	NN	O	O
ways	NN	O	O
to	NN	O	O
control	NN	O	O
inappropriate	NN	O	O
apoptosis	NN	O	O
involved	NN	O	O
in	NN	O	O
several	NN	O	O
diseases	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cross-priming	NN	O	O
of	NN	O	O
CTL	NN	O	O
responses	NN	O	O
in	NN	O	O
vivo	NN	O	O
does	NN	O	O
not	NN	O	O
require	NN	O	O
antigenic	NN	O	O
peptides	NN	O	O
in	NN	O	O
the	NN	O	O
endoplasmic	NN	O	O
reticulum	NN	O	O
of	NN	O	O
immunizing	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
proposed	NN	O	O
that	NN	O	O
the	NN	O	O
cross-priming	NN	O	O
of	NN	O	O
CTL	NN	O	O
responses	NN	O	O
in	NN	O	O
vivo	NN	O	O
involves	NN	O	O
the	NN	O	O
transfer	NN	O	O
to	NN	O	O
host	NN	O	O
APCs	NN	O	B-cell_type
of	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
protein	NN	O	I-protein
glycoprotein	NN	O	I-protein
96	NN	O	I-protein
-chaperoned	NN	O	O
antigenic	NN	O	O
peptides	NN	O	O
released	NN	O	O
from	NN	O	O
the	NN	O	O
endoplasmic	NN	O	O
reticulum	NN	O	O
(	NN	O	O
ER	NN	O	O
)	NN	O	O
of	NN	O	O
dying	NN	O	B-cell_type
or	NN	O	I-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
tested	NN	O	O
this	NN	O	O
possibility	NN	O	O
directly	NN	O	O
using	NN	O	O
TAP-deficient	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
lacking	NN	O	O
antigenic	NN	O	O
ER	NN	O	O
peptides	NN	O	O
derived	NN	O	O
from	NN	O	O
two	NN	O	O
model	NN	O	O
Ags	NN	O	B-protein
,	NN	O	O
the	NN	O	O
human	NN	O	B-protein
adenovirus	NN	O	I-protein
type	NN	O	I-protein
5	NN	O	I-protein
early	NN	O	I-protein
regions	NN	O	I-protein
E1A	NN	O	I-protein
and	NN	O	I-protein
E1B	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
both	NN	O	O
proteins	NN	O	O
were	NN	O	O
well	NN	O	O
expressed	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
not	NN	O	O
recognized	NN	O	O
by	NN	O	O
E1A-	NN	O	B-cell_type
or	NN	O	I-cell_type
E1B-specific	NN	O	I-cell_type
CTLs	NN	O	I-cell_type
unless	NN	O	O
the	NN	O	O
relevant	NN	O	O
epitope	NN	O	O
was	NN	O	O
either	NN	O	O
provided	NN	O	O
exogenously	NN	O	O
as	NN	O	O
a	NN	O	O
synthetic	NN	O	O
peptide	NN	O	O
or	NN	O	O
targeted	NN	O	O
to	NN	O	O
the	NN	O	O
ER	NN	O	O
in	NN	O	O
a	NN	O	O
TAP-independent	NN	O	O
fashion	NN	O	O
.	NN	O	O

Despite	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
these	NN	O	O
ER	NN	O	O
peptides	NN	O	O
,	NN	O	O
the	NN	O	O
TAP1-/-	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
able	NN	O	O
to	NN	O	O
efficiently	NN	O	O
cross-prime	NN	O	B-cell_type
E1A-	NN	O	I-cell_type
and	NN	O	I-cell_type
E1B-specific	NN	O	I-cell_type
CTLs	NN	O	I-cell_type
following	NN	O	O
immunization	NN	O	O
of	NN	O	O
syngeneic	NN	O	O
mice	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
,	NN	O	O
although	NN	O	O
purified	NN	O	O
peptide/glycoprotein	NN	O	B-protein
96	NN	O	I-protein
complexes	NN	O	I-protein
are	NN	O	O
potent	NN	O	O
immunogens	NN	O	O
,	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
CTL	NN	O	O
cross-priming	NN	O	O
in	NN	O	O
vivo	NN	O	O
does	NN	O	O
not	NN	O	O
depend	NN	O	O
upon	NN	O	O
antigenic	NN	O	O
peptides	NN	O	O
in	NN	O	O
the	NN	O	O
ER	NN	O	O
of	NN	O	O
immunizing	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
critical	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
p75	NN	O	B-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
p75TNF-R	NN	O	B-protein
)	NN	O	O
in	NN	O	O
organ	NN	O	O
inflammation	NN	O	O
independent	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
lymphotoxin	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
or	NN	O	O
the	NN	O	O
p55TNF-R	NN	O	B-protein
.	NN	O	I-protein

Despite	NN	O	O
overwhelming	NN	O	O
evidence	NN	O	O
that	NN	O	O
enhanced	NN	O	O
production	NN	O	O
of	NN	O	O
the	NN	O	O
p75	NN	O	B-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
p75TNF-R	NN	O	B-protein
)	NN	O	O
accompanies	NN	O	O
development	NN	O	O
of	NN	O	O
specific	NN	O	O
human	NN	O	O
inflammatory	NN	O	O
pathologies	NN	O	O
such	NN	O	O
as	NN	O	O
multi-organ	NN	O	O
failure	NN	O	O
during	NN	O	O
sepsis	NN	O	O
,	NN	O	O
inflammatory	NN	O	O
liver	NN	O	O
disease	NN	O	O
,	NN	O	O
pancreatitis	NN	O	O
,	NN	O	O
respiratory	NN	O	O
distress	NN	O	O
syndrome	NN	O	O
,	NN	O	O
or	NN	O	O
AIDS	NN	O	O
,	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
this	NN	O	O
receptor	NN	O	O
remains	NN	O	O
poorly	NN	O	O
defined	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
at	NN	O	O
levels	NN	O	O
relevant	NN	O	O
to	NN	O	O
human	NN	O	O
disease	NN	O	O
,	NN	O	O
production	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
p75TNF-R	NN	O	I-protein
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
severe	NN	O	O
inflammatory	NN	O	O
syndrome	NN	O	O
involving	NN	O	O
mainly	NN	O	O
the	NN	O	O
pancreas	NN	O	O
,	NN	O	O
liver	NN	O	O
,	NN	O	O
kidney	NN	O	O
,	NN	O	O
and	NN	O	O
lung	NN	O	O
,	NN	O	O
and	NN	O	O
characterized	NN	O	O
by	NN	O	O
constitutively	NN	O	O
increased	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cell	NN	O	I-cell_type
compartment	NN	O	O
.	NN	O	O

This	NN	O	O
process	NN	O	O
is	NN	O	O
shown	NN	O	O
to	NN	O	O
evolve	NN	O	O
independently	NN	O	O
of	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
lymphotoxin	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
or	NN	O	O
the	NN	O	O
p55TNF-R	NN	O	B-protein
,	NN	O	O
although	NN	O	O
coexpression	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
TNF	NN	O	I-DNA
transgene	NN	O	I-DNA
accelerated	NN	O	O
pathology	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
establish	NN	O	O
an	NN	O	O
independent	NN	O	O
role	NN	O	O
for	NN	O	O
enhanced	NN	O	O
p75TNF-R	NN	O	B-protein
production	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
disease	NN	O	O
and	NN	O	O
implicate	NN	O	O
the	NN	O	O
direct	NN	O	O
involvement	NN	O	O
of	NN	O	O
this	NN	O	O
receptor	NN	O	O
in	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
human	NN	O	O
inflammatory	NN	O	O
pathologies	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
status	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
and	NN	O	O
syndecan-1	NN	O	B-protein
identifies	NN	O	O
distinct	NN	O	O
histogenetic	NN	O	O
subtypes	NN	O	O
of	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

The	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
most	NN	O	O
cases	NN	O	O
of	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
(	NN	O	O
HD	NN	O	O
)	NN	O	O
have	NN	O	O
been	NN	O	O
recently	NN	O	O
recognized	NN	O	O
to	NN	O	O
originate	NN	O	O
from	NN	O	O
the	NN	O	O
B-cell	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
their	NN	O	O
precise	NN	O	O
differentiation	NN	O	O
stage	NN	O	O
is	NN	O	O
not	NN	O	O
fully	NN	O	O
clarified	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
reported	NN	O	O
that	NN	O	O
the	NN	O	O
histogenesis	NN	O	O
of	NN	O	O
B-cell	NN	O	B-cell_type
lymphomas	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
assessed	NN	O	O
by	NN	O	O
monitoring	NN	O	O
the	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
BCL-6	NN	O	B-protein
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
expressed	NN	O	O
in	NN	O	O
germinal	NN	O	B-cell_type
center	NN	O	I-cell_type
(	NN	O	I-cell_type
GC	NN	O	I-cell_type
)	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
CD138/syndecan-1	NN	O	B-protein
(	NN	O	O
syn-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
proteoglycan	NN	O	O
associated	NN	O	O
with	NN	O	O
post-GC	NN	O	O
,	NN	O	O
terminal	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
applied	NN	O	O
these	NN	O	O
two	NN	O	O
markers	NN	O	O
to	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
HD	NN	O	O
histogenesis	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
in	NN	O	O
nodular	NN	O	O
lymphocyte	NN	O	O
predominance	NN	O	O
HD	NN	O	O
(	NN	O	O
NLPHD	NN	O	O
)	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
consistently	NN	O	O
display	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
(	NN	O	O
+	NN	O	O
)	NN	O	O
/	NN	O	O
syn-1	NN	O	B-protein
(	NN	O	O
-	NN	O	O
)	NN	O	O
phenotype	NN	O	O
,	NN	O	O
indicating	NN	O	O
their	NN	O	O
derivation	NN	O	O
from	NN	O	O
GC	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
classic	NN	O	O
HD	NN	O	O
(	NN	O	O
CHD	NN	O	O
)	NN	O	O
is	NN	O	O
heterogeneous	NN	O	O
because	NN	O	O
the	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
a	NN	O	O
fraction	NN	O	O
of	NN	O	O
CHD	NN	O	O
display	NN	O	O
the	NN	O	O
BCL-6	NN	O	B-protein
(	NN	O	O
-	NN	O	O
)	NN	O	O
/	NN	O	O
syn-1	NN	O	B-protein
(	NN	O	O
+	NN	O	O
)	NN	O	O
phenotype	NN	O	O
of	NN	O	O
post-GC	NN	O	O
B-cells	NN	O	O
,	NN	O	O
whereas	NN	O	O
another	NN	O	O
fraction	NN	O	O
of	NN	O	O
CHD	NN	O	O
is	NN	O	O
constituted	NN	O	O
by	NN	O	O
a	NN	O	O
mixture	NN	O	O
of	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
reflecting	NN	O	O
the	NN	O	O
GC	NN	O	O
(	NN	O	O
BCL-6	NN	O	B-protein
(	NN	O	O
+	NN	O	O
)	NN	O	O
/	NN	O	O
syn-1	NN	O	B-protein
(	NN	O	O
-	NN	O	O
)	NN	O	O
)	NN	O	O
or	NN	O	O
post-GC	NN	O	O
(	NN	O	O
BCL-6	NN	O	B-protein
(	NN	O	O
-	NN	O	O
)	NN	O	O
/	NN	O	O
syn-1	NN	O	B-protein
(	NN	O	O
+	NN	O	O
)	NN	O	O
)	NN	O	O
phenotypes	NN	O	O
.	NN	O	O

BCL-6	NN	O	B-cell_line
(	NN	O	I-cell_line
-	NN	O	I-cell_line
)	NN	O	I-cell_line
/syn-1	NN	O	I-cell_line
(	NN	O	I-cell_line
+	NN	O	I-cell_line
)	NN	O	I-cell_line
tumor	NN	O	I-cell_line
cells	NN	O	I-cell_line
of	NN	O	O
CHD	NN	O	O
are	NN	O	O
mostly	NN	O	O
found	NN	O	O
surrounded	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
expressing	NN	O	O
CD40L	NN	O	B-protein
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
CD40	NN	O	B-protein
signaling	NN	O	O
downregulates	NN	O	O
BCL-6	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
NLPHD	NN	O	O
uniformly	NN	O	O
display	NN	O	O
a	NN	O	O
GC	NN	O	O
B-cell	NN	O	O
phenotype	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
phenotype	NN	O	O
of	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
CHD	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
modulated	NN	O	O
by	NN	O	O
the	NN	O	O
surrounding	NN	O	O
cellular	NN	O	O
background	NN	O	O
,	NN	O	O
particularly	NN	O	O
CD40L+	NN	O	B-cell_type
reactive	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
and	NN	O	O
cellular	NN	O	O
analysis	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
lymphoblastoid	NN	O	B-protein
T-cell-line-expressing	NN	O	I-protein
wild-type	NN	O	I-protein
and	NN	O	I-protein
mutated	NN	O	I-protein
CD4	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
cytoplasmic	NN	O	I-protein
tail	NN	O	I-protein
is	NN	O	O
critical	NN	O	O
for	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
-induced	NN	O	O
apoptosis	NN	O	O
(	NN	O	O
J.Corbeil	NN	O	O
,	NN	O	O
M.Tremblay	NN	O	O
,	NN	O	O
and	NN	O	O
D.D.Richman	NN	O	O
,	NN	O	O
J.Exp.Med.183	NN	O	O
:	NN	O	O
39-48	NN	O	O
,	NN	O	O
1996	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
pursued	NN	O	O
our	NN	O	O
investigation	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
transduction	NN	O	O
pathway	NN	O	O
in	NN	O	O
HIV-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

To	NN	O	O
do	NN	O	O
this	NN	O	O
,	NN	O	O
wild-type	NN	O	O
and	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
cytoplasmic	NN	O	I-protein
tail	NN	O	I-protein
were	NN	O	O
stably	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
lymphoblastoid	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
A2.01	NN	O	B-cell_line
.	NN	O	O

Apoptosis	NN	O	O
was	NN	O	O
prevented	NN	O	O
when	NN	O	O
CD4	NN	O	B-protein
truncated	NN	O	O
at	NN	O	O
residue	NN	O	O
402	NN	O	O
was	NN	O	O
expressed	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
cells	NN	O	O
expressing	NN	O	O
mutated	NN	O	O
receptors	NN	O	O
that	NN	O	O
do	NN	O	O
not	NN	O	O
associate	NN	O	O
with	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
(	NN	O	O
mutated	NN	O	O
at	NN	O	O
the	NN	O	O
dicysteine	NN	O	B-protein
motif	NN	O	I-protein
and	NN	O	O
truncated	NN	O	O
at	NN	O	O
residue	NN	O	O
418	NN	O	O
)	NN	O	O
but	NN	O	O
which	NN	O	O
conserved	NN	O	O
proximal	NN	O	O
domains	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
tail	NN	O	I-protein
underwent	NN	O	O
apoptosis	NN	O	O
like	NN	O	O
wild-	NN	O	B-protein
type	NN	O	I-protein
CD4	NN	O	I-protein
.	NN	O	O

The	NN	O	O
differences	NN	O	O
between	NN	O	O
wild-type	NN	O	B-protein
and	NN	O	I-protein
mutated	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
were	NN	O	O
not	NN	O	O
related	NN	O	O
to	NN	O	O
levels	NN	O	O
of	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
or	NN	O	O
NF-	NN	O	B-protein
kappaB	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Initial	NN	O	O
signaling	NN	O	O
through	NN	O	O
the	NN	O	O
CD4	NN	O	B-protein
receptor	NN	O	I-protein
played	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
sensitization	NN	O	O
of	NN	O	O
HIV-infected	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
undergo	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
HIV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
(	NN	O	I-protein
MAb	NN	O	I-protein
)	NN	O	I-protein
13B8-2	NN	O	I-protein
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
CD4	NN	O	B-protein
in	NN	O	O
a	NN	O	O
region	NN	O	O
critical	NN	O	O
for	NN	O	O
dimerization	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
,	NN	O	O
prevented	NN	O	O
apoptosis	NN	O	O
without	NN	O	O
inhibiting	NN	O	O
HIV	NN	O	O
replication	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
apoptotic	NN	O	O
process	NN	O	O
was	NN	O	O
not	NN	O	O
related	NN	O	O
to	NN	O	O
Fas	NN	O	B-protein
-Fas	NN	O	B-protein
ligand	NN	O	I-protein
interaction	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
an	NN	O	O
antagonistic	NN	O	O
anti-Fas	NN	O	O
MAb	NN	O	O
(	NN	O	O
ZB-4	NN	O	O
)	NN	O	O
enhanced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
HIV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
without	NN	O	O
inducing	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
uninfected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
observations	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
CD4	NN	O	B-protein
signaling	NN	O	O
mediates	NN	O	O
HIV-induced	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
a	NN	O	O
mechanism	NN	O	O
independent	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
-Fas	NN	O	B-protein
ligand	NN	O	I-protein
interaction	NN	O	O
,	NN	O	O
does	NN	O	O
not	NN	O	O
require	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
signaling	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
involve	NN	O	O
a	NN	O	O
critical	NN	O	O
region	NN	O	O
for	NN	O	O
CD4	NN	O	B-protein
dimerization	NN	O	O
.	NN	O	O

-DOCSTART-	O

p130	NN	O	B-protein
,	NN	O	O
p107	NN	O	B-protein
,	NN	O	O
and	NN	O	O
pRb	NN	O	B-protein
are	NN	O	O
differentially	NN	O	O
regulated	NN	O	O
in	NN	O	O
proliferating	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
during	NN	O	O
cell	NN	O	O
cycle	NN	O	O
arrest	NN	O	O
by	NN	O	O
alpha-interferon	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
determined	NN	O	O
how	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-protein
family	NN	O	I-protein
(	NN	O	O
pRb	NN	O	B-protein
,	NN	O	O
p107	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p130	NN	O	B-protein
)	NN	O	O
is	NN	O	O
governed	NN	O	O
in	NN	O	O
individual	NN	O	O
cell	NN	O	O
cycle	NN	O	O
phases	NN	O	O
of	NN	O	O
Daudi	NN	O	B-cell_line
B-cells	NN	O	I-cell_line
during	NN	O	O
cell	NN	O	O
cycle	NN	O	O
exit	NN	O	O
triggered	NN	O	O
by	NN	O	O
alpha-interferon	NN	O	B-protein
(	NN	O	O
alpha-IFN	NN	O	B-protein
)	NN	O	O
.	NN	O	O

alpha-IFN	NN	O	B-protein
causes	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
pRb	NN	O	B-protein
and	NN	O	O
loss	NN	O	O
of	NN	O	O
p130	NN	O	B-protein
phosphorylated	NN	O	I-protein
Form	NN	O	I-protein
3	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
change	NN	O	O
in	NN	O	O
p130	NN	O	B-protein
phosphorylation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
alpha-IFN	NN	O	B-protein
occurs	NN	O	O
before	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
pRb	NN	O	B-protein
is	NN	O	O
complete	NN	O	O
because	NN	O	O
loss	NN	O	O
of	NN	O	O
p130	NN	O	B-protein
Form	NN	O	I-protein
3	NN	O	I-protein
occurs	NN	O	O
throughout	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
prior	NN	O	O
to	NN	O	O
complete	NN	O	O
arrest	NN	O	O
in	NN	O	O
G1	NN	O	O
,	NN	O	O
whereas	NN	O	O
pRb	NN	O	B-protein
is	NN	O	O
dephosphorylated	NN	O	O
only	NN	O	O
in	NN	O	O
G1	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
p107	NN	O	B-protein
is	NN	O	O
dephosphorylated	NN	O	O
and	NN	O	O
is	NN	O	O
then	NN	O	O
depleted	NN	O	O
from	NN	O	O
cells	NN	O	O
as	NN	O	O
they	NN	O	O
exit	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

p130	NN	O	B-protein
,	NN	O	O
predominantly	NN	O	O
in	NN	O	O
Form	NN	O	O
1	NN	O	O
,	NN	O	O
and	NN	O	O
hypophosphorylated	NN	O	B-protein
pRb	NN	O	I-protein
bind	NN	O	O
an	NN	O	O
E2F	NN	O	B-DNA
DNA	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
;	NN	O	O
p130	NN	O	B-protein
complexes	NN	O	O
E2F-4	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
pRb	NN	O	B-protein
binds	NN	O	O
both	NN	O	O
E2F-4	NN	O	O
and	NN	O	O
E2F-1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
phosphorylated	NN	O	O
forms	NN	O	O
of	NN	O	O
E2F-4	NN	O	B-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
E2F	NN	O	B-DNA
DNA	NN	O	I-DNA
site	NN	O	I-DNA
are	NN	O	O
different	NN	O	O
from	NN	O	O
hyperphosphorylated	NN	O	O
E2F-4	NN	O	B-protein
,	NN	O	O
which	NN	O	O
predominates	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
hemopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
G0	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
although	NN	O	O
cell	NN	O	O
cycle	NN	O	O
arrest	NN	O	O
induced	NN	O	O
by	NN	O	O
alpha-IFN	NN	O	B-protein
may	NN	O	O
be	NN	O	O
mediated	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
complex	NN	O	O
containing	NN	O	O
p130	NN	O	B-protein
and	NN	O	O
E2F-4	NN	O	B-protein
,	NN	O	O
alpha-IFN	NN	O	B-protein
does	NN	O	O
not	NN	O	O
induce	NN	O	O
hyperphosphorylation	NN	O	O
of	NN	O	O
E2F-4	NN	O	B-protein
,	NN	O	O
which	NN	O	O
characterizes	NN	O	O
primary	NN	O	B-cell_type
hemopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
G0	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
c-myb	NN	O	B-protein
and	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
regulate	NN	O	O
the	NN	O	O
monocytic/myeloic	NN	O	B-DNA
gene	NN	O	I-DNA
MRP14	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
entry	NN	O	O
of	NN	O	O
microorganisms	NN	O	O
into	NN	O	O
the	NN	O	O
body	NN	O	O
induces	NN	O	O
inflammatory	NN	O	O
processes	NN	O	O
.	NN	O	O

During	NN	O	O
this	NN	O	O
process	NN	O	O
a	NN	O	O
sequence	NN	O	O
of	NN	O	O
cellular	NN	O	O
,	NN	O	O
humoral	NN	O	O
,	NN	O	O
non-specific	NN	O	O
and	NN	O	O
specific	NN	O	O
actions	NN	O	O
are	NN	O	O
evoked	NN	O	O
to	NN	O	O
combat	NN	O	O
the	NN	O	O
infection	NN	O	O
.	NN	O	O

Macrophages	NN	O	B-cell_type
and	NN	O	O
granulocytes	NN	O	B-cell_type
,	NN	O	O
which	NN	O	O
are	NN	O	O
developed	NN	O	O
from	NN	O	O
a	NN	O	O
common	NN	O	O
progenitor	NN	O	O
cell	NN	O	O
,	NN	O	O
are	NN	O	O
the	NN	O	O
cellular	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
specific	NN	O	O
and	NN	O	O
non-specific	NN	O	O
immunoreaction	NN	O	O
.	NN	O	O

MRP14	NN	O	B-DNA
(	NN	O	O
Macrophage	NN	O	B-protein
migration	NN	O	I-protein
inhibitory	NN	O	I-protein
related	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
and	NN	O	O
MRP8	NN	O	B-protein
,	NN	O	O
two	NN	O	O
S-100	NN	O	B-protein
proteins	NN	O	I-protein
contained	NN	O	O
in	NN	O	O
high	NN	O	O
concentrations	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
are	NN	O	O
obviously	NN	O	O
essential	NN	O	O
for	NN	O	O
adhesion	NN	O	O
and	NN	O	O
migration	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
granulocytes	NN	O	B-cell_type
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
we	NN	O	O
cotransfected	NN	O	O
constructs	NN	O	O
expressing	NN	O	O
CAT	NN	O	B-protein
under	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
MRP14	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
expression	NN	O	O
constructs	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
v-myb	NN	O	B-protein
,	NN	O	O
two	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
myeloid/monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Transfection	NN	O	O
with	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
revealed	NN	O	O
a	NN	O	O
massive	NN	O	O
enhancement	NN	O	O
of	NN	O	O
the	NN	O	O
MRP14	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
both	NN	O	O
,	NN	O	O
HL	NN	O	B-cell_type
60	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
granulocytic	NN	O	O
differentiated	NN	O	O
)	NN	O	O
and	NN	O	O
L132	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
v-myb	NN	O	B-protein
reduces	NN	O	O
MRP14	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
L132	NN	O	B-cell_type
cells	NN	O	I-cell_type
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
vector	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
enhance	NN	O	O
MRP14	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
whole	NN	O	O
genome	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
CD28	NN	O	B-protein
-associated	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
leading	NN	O	O
to	NN	O	O
cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
without	NN	O	O
TCR	NN	O	B-protein
engagement	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
CD28-specific	NN	O	B-protein
mAb	NN	O	I-protein
BW	NN	O	I-protein
828	NN	O	I-protein
induces	NN	O	O
proliferation	NN	O	O
and	NN	O	O
cytokine	NN	O	B-protein
synthesis	NN	O	O
without	NN	O	O
further	NN	O	O
requirement	NN	O	O
for	NN	O	O
TCR	NN	O	B-protein
coengagement	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
prompted	NN	O	O
us	NN	O	O
to	NN	O	O
postulate	NN	O	O
that	NN	O	O
signal	NN	O	O
2	NN	O	O
(	NN	O	O
costimulatory	NN	O	O
signal	NN	O	O
)	NN	O	O
alone	NN	O	O
without	NN	O	O
signal	NN	O	O
1	NN	O	O
(	NN	O	O
TCR	NN	O	B-protein
signal	NN	O	O
)	NN	O	O
can	NN	O	O
activate	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
test	NN	O	O
whether	NN	O	O
this	NN	O	O
putative	NN	O	O
function	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
is	NN	O	O
mediated	NN	O	O
via	NN	O	O
a	NN	O	O
particular	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
,	NN	O	O
we	NN	O	O
compared	NN	O	O
early	NN	O	O
signaling	NN	O	O
events	NN	O	O
initiated	NN	O	O
in	NN	O	O
resting	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
the	NN	O	O
stimulatory	NN	O	B-protein
mAb	NN	O	I-protein
BW	NN	O	I-protein
828	NN	O	I-protein
with	NN	O	O
signals	NN	O	O
triggered	NN	O	O
by	NN	O	O
the	NN	O	O
nonstimulating	NN	O	B-protein
CD28	NN	O	I-protein
mAb	NN	O	I-protein
9.3	NN	O	I-protein
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
BW	NN	O	B-protein
828	NN	O	I-protein
induced	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
,	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
lead	NN	O	O
to	NN	O	O
detectable	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinases	NN	O	I-protein
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
c-Raf-1	NN	O	B-protein
.	NN	O	O

This	NN	O	O
pathway	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
and	NN	O	O
proteins	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
CD28	NN	O	B-protein
response	NN	O	O
element	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
mAb	NN	O	B-protein
9.3	NN	O	I-protein
increased	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
and	NN	O	O
triggered	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
c-Raf-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
was	NN	O	O
unable	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
differential	NN	O	O
signaling	NN	O	O
of	NN	O	O
BW	NN	O	B-protein
828	NN	O	I-protein
and	NN	O	O
9.3	NN	O	B-protein
in	NN	O	O
resting	NN	O	O
T	NN	O	O
cells	NN	O	O
,	NN	O	O
the	NN	O	O
two	NN	O	O
mAbs	NN	O	O
exhibited	NN	O	O
a	NN	O	O
similar	NN	O	O
pattern	NN	O	O
of	NN	O	O
early	NN	O	O
signaling	NN	O	O
events	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
Jurkat	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
p56	NN	O	B-protein
(	NN	O	I-protein
lck	NN	O	I-protein
)	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
association	NN	O	O
of	NN	O	O
phosphatidylinositol	NN	O	B-protein
3-kinase	NN	O	I-protein
with	NN	O	O
CD28	NN	O	B-protein
)	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
signaling	NN	O	O
capacity	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
changes	NN	O	O
with	NN	O	O
activation	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
view	NN	O	O
that	NN	O	O
stimulation	NN	O	O
through	NN	O	O
CD28	NN	O	B-protein
can	NN	O	O
induce	NN	O	O
some	NN	O	O
effector	NN	O	O
functions	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
this	NN	O	O
capacity	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
particular	NN	O	O
pattern	NN	O	O
of	NN	O	O
early	NN	O	O
signaling	NN	O	O
events	NN	O	O

-DOCSTART-	O

Decreased	NN	O	O
IL-12	NN	O	B-protein
production	NN	O	O
and	NN	O	O
Th1	NN	O	O
cell	NN	O	O
development	NN	O	O
by	NN	O	O
acetyl	NN	O	O
salicylic	NN	O	O
acid-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

IL-12	NN	O	B-protein
is	NN	O	O
a	NN	O	O
75-kDa	NN	O	B-protein
heterodimeric	NN	O	I-protein
cytokine	NN	O	I-protein
composed	NN	O	O
of	NN	O	O
two	NN	O	O
covalently	NN	O	O
linked	NN	O	O
p35	NN	O	B-protein
and	NN	O	I-protein
p40	NN	O	I-protein
chains	NN	O	I-protein
.	NN	O	O

This	NN	O	O
pro-inflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
prominent	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
Th1	NN	O	O
cell-mediated	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

Th1	NN	O	O
cell-mediated	NN	O	O
immune	NN	O	O
responses	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
chronic	NN	O	O
inflammatory	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
IL-12	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
critical	NN	O	O
factor	NN	O	O
in	NN	O	O
the	NN	O	O
generation	NN	O	O
and	NN	O	O
maintenance	NN	O	O
of	NN	O	O
chronic	NN	O	O
inflammatory	NN	O	O
conditions	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
a	NN	O	O
commonly	NN	O	O
prescribed	NN	O	O
anti-inflammatory	NN	O	O
drug	NN	O	O
,	NN	O	O
acetyl	NN	O	O
salicylic	NN	O	O
acid	NN	O	O
(	NN	O	O
ASA	NN	O	O
)	NN	O	O
,	NN	O	O
on	NN	O	O
IL-12	NN	O	B-protein
production	NN	O	O
and	NN	O	O
Th1	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

ASA	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
inhibit	NN	O	O
secretion	NN	O	O
of	NN	O	O
the	NN	O	O
IL-12	NN	O	B-protein
heterodimer	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
p40	NN	O	B-protein
monomer	NN	O	I-protein
by	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
IL-12p40	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
p40	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
revealed	NN	O	O
that	NN	O	O
ASA	NN	O	O
inhibited	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
p40-kappaB	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
p40	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
leading	NN	O	O
to	NN	O	O
transcriptional	NN	O	O
repression	NN	O	O
of	NN	O	O
the	NN	O	O
p40	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
ASA	NN	O	O
to	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
T	NN	O	O
helper	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
system	NN	O	O
,	NN	O	O
at	NN	O	O
concentrations	NN	O	O
compatible	NN	O	O
with	NN	O	O
plasma	NN	O	O
levels	NN	O	O
reached	NN	O	O
during	NN	O	O
anti-inflammatory	NN	O	O
therapy	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
reduced	NN	O	O
development	NN	O	O
of	NN	O	O
Th1	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
ASA	NN	O	O
leads	NN	O	O
to	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
production	NN	O	O
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Th1	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Attenuation	NN	O	O
of	NN	O	O
HLA-DR	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
infected	NN	O	O
with	NN	O	O
Mycobacterium	NN	O	O
tuberculosis	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
intracellular	NN	O	O
sequestration	NN	O	O
of	NN	O	O
immature	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
heterodimers	NN	O	I-protein
.	NN	O	O

MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
infected	NN	O	O
with	NN	O	O
Mycobacterium	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

IFN-gamma	NN	O	B-protein
increased	NN	O	O
the	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
this	NN	O	O
was	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
in	NN	O	O
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

Despite	NN	O	O
this	NN	O	O
effect	NN	O	O
,	NN	O	O
steady	NN	O	O
state	NN	O	O
levels	NN	O	O
of	NN	O	O
HLA-DRalpha	NN	O	B-protein
,	NN	O	O
HLA-DRbeta	NN	O	B-protein
,	NN	O	O
and	NN	O	O
invariant	NN	O	B-protein
(	NN	O	I-protein
Ii	NN	O	I-protein
)	NN	O	I-protein
chains	NN	O	I-protein
were	NN	O	O
equivalent	NN	O	O
in	NN	O	O
control	NN	O	B-cell_type
and	NN	O	I-cell_type
infected	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Metabolic	NN	O	O
labeling	NN	O	O
combined	NN	O	O
with	NN	O	O
pulse-chase	NN	O	O
experiments	NN	O	O
and	NN	O	O
biochemical	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
infected	NN	O	O
cells	NN	O	O
became	NN	O	O
resistant	NN	O	O
to	NN	O	O
endoglycosidase	NN	O	B-protein
H	NN	O	I-protein
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
normal	NN	O	O
Golgi	NN	O	O
processing	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
results	NN	O	O
of	NN	O	O
intracellular	NN	O	O
staining	NN	O	O
and	NN	O	O
dual	NN	O	O
color	NN	O	O
confocal	NN	O	O
microscopy	NN	O	O
revealed	NN	O	O
a	NN	O	O
significant	NN	O	O
defect	NN	O	O
in	NN	O	O
transport	NN	O	O
of	NN	O	O
newly	NN	O	O
synthesized	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
through	NN	O	O
the	NN	O	O
endocytic	NN	O	O
compartment	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
compared	NN	O	O
with	NN	O	O
findings	NN	O	O
in	NN	O	O
control	NN	O	O
cells	NN	O	O
,	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
colocalized	NN	O	O
to	NN	O	O
a	NN	O	O
minimal	NN	O	O
extent	NN	O	O
with	NN	O	O
a	NN	O	O
lysosomal-associated	NN	O	B-protein
membrane	NN	O	I-protein
protein-1	NN	O	I-protein
+	NN	O	O
endosomal	NN	O	O
compartment	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
control	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
failed	NN	O	O
to	NN	O	O
colocalize	NN	O	O
with	NN	O	O
endocytosed	NN	O	B-protein
BSA	NN	O	I-protein
under	NN	O	O
conditions	NN	O	O
where	NN	O	O
this	NN	O	O
marker	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
label	NN	O	O
late	NN	O	O
endosomes	NN	O	O
,	NN	O	O
lysosomes	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
compartment	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
defective	NN	O	O
transport	NN	O	O
along	NN	O	O
the	NN	O	O
endocytic	NN	O	O
pathway	NN	O	O
,	NN	O	O
the	NN	O	O
maturation	NN	O	O
of	NN	O	O
SDS-stable	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
alphabeta	NN	O	I-protein
dimers	NN	O	I-protein
--	NN	O	O
dependent	NN	O	O
upon	NN	O	O
removal	NN	O	O
of	NN	O	O
Ii	NN	O	B-protein
chain	NN	O	I-protein
and	NN	O	O
peptide	NN	O	O
loading	NN	O	O
of	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
dimers	NN	O	I-protein
in	NN	O	O
the	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
compartment	NN	O	O
--	NN	O	O
was	NN	O	O
markedly	NN	O	O
impaired	NN	O	O
in	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
defective	NN	O	O
transport	NN	O	O
and	NN	O	O
processing	NN	O	O
of	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
through	NN	O	O
the	NN	O	O
endosomal/lysosomal	NN	O	O
system	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
diminished	NN	O	O
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
that	NN	O	O
repress	NN	O	O
expression	NN	O	O
of	NN	O	O
adult	NN	O	B-DNA
beta-like	NN	O	I-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
primitive	NN	O	O
erythroid	NN	O	O
environment	NN	O	O
.	NN	O	O

Our	NN	O	O
investigations	NN	O	O
have	NN	O	O
focused	NN	O	O
on	NN	O	O
localizing	NN	O	O
cis-elements	NN	O	B-DNA
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
down	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
adult	NN	O	B-DNA
beta-like	NN	O	I-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
(	NN	O	O
delta	NN	O	B-DNA
and	NN	O	O
beta	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
immature	NN	O	O
,	NN	O	O
or	NN	O	O
primitive	NN	O	O
erythroid	NN	O	O
tissues	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
their	NN	O	O
activity	NN	O	O
after	NN	O	O
transfection	NN	O	O
into	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
an	NN	O	O
erythroleukemia	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
with	NN	O	O
an	NN	O	O
embryonic-fetal	NN	O	O
phenotype	NN	O	O
.	NN	O	O

Analyzed	NN	O	O
DNA	NN	O	B-DNA
sequences	NN	O	I-DNA
included	NN	O	O
delta	NN	O	B-DNA
and	NN	O	I-DNA
beta	NN	O	I-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
regions	NN	O	I-DNA
extending	NN	O	O
from	NN	O	O
approximately	NN	O	O
-500	NN	O	B-DNA
to	NN	O	I-DNA
+50bp	NN	O	I-DNA
(	NN	O	O
promoter	NN	O	B-DNA
regions	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
truncated	NN	O	O
delta	NN	O	O
and	NN	O	O
beta	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
regions	NN	O	I-DNA
extending	NN	O	O
from	NN	O	O
approximately	NN	O	O
-250	NN	O	B-DNA
to	NN	O	I-DNA
+50	NN	O	I-DNA
bp	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
chimeric	NN	O	B-DNA
promoter	NN	O	I-DNA
constructions	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
consisted	NN	O	O
of	NN	O	O
a	NN	O	O
distal	NN	O	O
delta	NN	O	B-DNA
or	NN	O	I-DNA
beta	NN	O	I-DNA
fragment	NN	O	I-DNA
fused	NN	O	O
to	NN	O	O
a	NN	O	O
proximal	NN	O	B-DNA
beta	NN	O	I-DNA
or	NN	O	I-DNA
delta	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
constructions	NN	O	I-DNA
no	NN	O	O
appreciable	NN	O	O
level	NN	O	O
of	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
supported	NN	O	O
by	NN	O	O
the	NN	O	O
beta	NN	O	B-DNA
globin	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
only	NN	O	O
low	NN	O	O
level	NN	O	O
activity	NN	O	O
by	NN	O	O
the	NN	O	O
delta	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Truncation	NN	O	O
of	NN	O	O
the	NN	O	O
beta	NN	O	B-DNA
globin	NN	O	I-DNA
promoter	NN	O	I-DNA
led	NN	O	O
to	NN	O	O
a	NN	O	O
2-3	NN	O	O
fold	NN	O	O
increase	NN	O	O
in	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
portion	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
delta	NN	O	B-DNA
promoter	NN	O	I-DNA
led	NN	O	O
to	NN	O	O
a	NN	O	O
10	NN	O	O
fold	NN	O	O
decrease	NN	O	O
in	NN	O	O
expression	NN	O	O
.	NN	O	O

Coupling	NN	O	O
of	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
beta	NN	O	I-DNA
globin	NN	O	I-DNA
sequence	NN	O	I-DNA
from	NN	O	O
approximately	NN	O	O
-500	NN	O	O
to	NN	O	O
-250	NN	O	O
bp	NN	O	O
to	NN	O	O
the	NN	O	O
truncated	NN	O	O
delta	NN	O	B-DNA
promoter	NN	O	I-DNA
fragment	NN	O	O
led	NN	O	O
to	NN	O	O
complete	NN	O	O
extinction	NN	O	O
of	NN	O	O
transcription	NN	O	O
activity	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
beta	NN	O	B-DNA
globin	NN	O	I-DNA
gene	NN	O	I-DNA
upstream	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
s	NN	O	O
)	NN	O	O
.	NN	O	O

Fusion	NN	O	O
of	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
portion	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
delta	NN	O	B-DNA
promoter	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
truncated	NN	O	O
beta	NN	O	B-DNA
globin	NN	O	I-DNA
promoter	NN	O	I-DNA
yielded	NN	O	O
a	NN	O	O
modest	NN	O	O
increase	NN	O	O
in	NN	O	O
promoter	NN	O	O
strength	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
truncated	NN	O	B-DNA
beta	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
positive	NN	O	B-DNA
transcriptional	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
s	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
delta	NN	O	I-DNA
globin	NN	O	I-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
repressor	NN	O	B-protein
proteins	NN	O	I-protein
BP1	NN	O	I-protein
and	NN	O	I-protein
BP2	NN	O	I-protein
in	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
portion	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
beta	NN	O	B-DNA
globin	NN	O	I-DNA
gene	NN	O	I-DNA
flanking	NN	O	I-DNA
region	NN	O	I-DNA
led	NN	O	O
to	NN	O	O
a	NN	O	O
4-6	NN	O	O
fold	NN	O	O
increase	NN	O	O
in	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

DNase	NN	O	B-protein
I	NN	O	I-protein
footprinting	NN	O	O
of	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
delta-globin	NN	O	I-DNA
region	NN	O	I-DNA
revealed	NN	O	O
protected	NN	O	B-DNA
sequences	NN	O	I-DNA
corresponding	NN	O	O
to	NN	O	O
consensus	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
GATA-1	NN	O	B-protein
and	NN	O	O
BP2	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
confirm	NN	O	O
that	NN	O	O
sequences	NN	O	O
in	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
adult	NN	O	B-DNA
beta	NN	O	I-DNA
globin	NN	O	I-DNA
gene	NN	O	I-DNA
contribute	NN	O	O
to	NN	O	O
its	NN	O	O
factor-mediated	NN	O	O
suppression	NN	O	O
early	NN	O	O
in	NN	O	O
development	NN	O	O
and	NN	O	O
then	NN	O	O
may	NN	O	O
modulate	NN	O	O
its	NN	O	O
expression	NN	O	O
at	NN	O	O
a	NN	O	O
later	NN	O	O
stage	NN	O	O
.	NN	O	O

-DOCSTART-	O

Analysis	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
extrinsic	NN	O	O
asthma	NN	O	O
and	NN	O	O
hyperimmunoglobulin	NN	O	O
E	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Recent	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
class	NN	O	O
switching	NN	O	O
to	NN	O	O
IgE	NN	O	B-protein
by	NN	O	O
cytokines	NN	O	B-protein
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
STAT	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-13	NN	O	B-protein
occurs	NN	O	O
through	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
intracellular	NN	O	B-protein
signal-transducing	NN	O	I-protein
protein	NN	O	I-protein
Stat6	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
class	NN	O	O
switching	NN	O	O
by	NN	O	O
interferon-y	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
occurs	NN	O	O
through	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Statl	NN	O	B-protein
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
We	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
in	NN	O	O
extrinsic	NN	O	O
asthma	NN	O	O
or	NN	O	O
in	NN	O	O
cases	NN	O	O
of	NN	O	O
markedly	NN	O	O
elevated	NN	O	O
IgE	NN	O	B-protein
(	NN	O	O
ie	NN	O	O
,	NN	O	O
hyperimmunoglobulin	NN	O	O
E	NN	O	O
[	NN	O	O
HIE	NN	O	O
]	NN	O	O
)	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
levels	NN	O	O
or	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
PBMCs	NN	O	B-cell_type
and	NN	O	O
sera	NN	O	O
from	NN	O	O
8	NN	O	O
patients	NN	O	O
with	NN	O	O
extrinsic	NN	O	O
asthma	NN	O	O
(	NN	O	O
mean	NN	O	O
IgE	NN	O	B-protein
,	NN	O	O
285+/-100	NN	O	O
IU/mL	NN	O	O
)	NN	O	O
,	NN	O	O
3	NN	O	O
patients	NN	O	O
with	NN	O	O
HIE	NN	O	O
(	NN	O	O
mean	NN	O	O
IgE	NN	O	B-protein
,	NN	O	O
7050+/-1122	NN	O	O
IU/mL	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
14	NN	O	O
nonatopic	NN	O	O
control	NN	O	O
subjects	NN	O	O
(	NN	O	O
mean	NN	O	O
IgE	NN	O	B-protein
,	NN	O	O
112+/-28	NN	O	O
IU/mL	NN	O	O
)	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
The	NN	O	O
mean	NN	O	O
IL-4	NN	O	B-protein
level	NN	O	O
detected	NN	O	O
by	NN	O	O
ELISA	NN	O	O
was	NN	O	O
much	NN	O	O
greater	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
HIE	NN	O	O
than	NN	O	O
control	NN	O	O
subjects	NN	O	O
(	NN	O	O
88.6+/-11.5	NN	O	O
pg/mL	NN	O	O
vs	NN	O	O
11.5+/-7.1	NN	O	O
pg/mL	NN	O	O
,	NN	O	O
P	NN	O	O
=	NN	O	O
.005	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
increased	NN	O	O
IL-4	NN	O	B-protein
levels	NN	O	O
among	NN	O	O
patients	NN	O	O
with	NN	O	O
both	NN	O	O
asthma	NN	O	O
and	NN	O	O
HIE	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
increased	NN	O	O
IgE	NN	O	B-protein
levels	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
IL-13	NN	O	B-protein
levels	NN	O	O
were	NN	O	O
not	NN	O	O
elevated	NN	O	O
.	NN	O	O

Levels	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
protein	NN	O	O
present	NN	O	O
in	NN	O	O
PBMCs	NN	O	B-cell_type
did	NN	O	O
not	NN	O	O
differ	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
and	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

Examination	NN	O	O
of	NN	O	O
Stat6	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
demonstrated	NN	O	O
no	NN	O	O
activation	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
either	NN	O	O
HIE	NN	O	O
or	NN	O	O
acute	NN	O	O
asthma	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
evidence	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
have	NN	O	O
already	NN	O	O
switched	NN	O	O
to	NN	O	O
IgE	NN	O	B-protein
was	NN	O	O
seen	NN	O	O
in	NN	O	O
PBMCs	NN	O	B-cell_type
of	NN	O	O
several	NN	O	O
patients	NN	O	O
with	NN	O	O
asthma	NN	O	O
or	NN	O	O
HIE	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
IgE	NN	O	B-protein
production	NN	O	O
in	NN	O	O
asthma	NN	O	O
and	NN	O	O
HIE	NN	O	O
usually	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
increased	NN	O	O
sera	NN	O	O
IL-4	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
asthma	NN	O	O
and	NN	O	O
HIE	NN	O	O
are	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
induce	NN	O	O
Stat6	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
PBMCs	NN	O	B-cell_type
;	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
evidence	NN	O	O
of	NN	O	O
switch	NN	O	O
recombination	NN	O	O
to	NN	O	O
epsilon	NN	O	O
may	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
isolated	NN	O	O
cases	NN	O	O
of	NN	O	O
elevated	NN	O	O
IgE	NN	O	B-protein
.	NN	O	O

This	NN	O	O
implies	NN	O	O
that	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
in	NN	O	O
these	NN	O	O
patients	NN	O	O
either	NN	O	O
results	NN	O	O
from	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
have	NN	O	O
already	NN	O	O
undergone	NN	O	O
class	NN	O	O
switching	NN	O	O
,	NN	O	O
from	NN	O	O
Ig	NN	O	O
class	NN	O	O
switching	NN	O	O
that	NN	O	O
is	NN	O	O
localized	NN	O	O
to	NN	O	O
target	NN	O	O
tissues	NN	O	O
,	NN	O	O
or	NN	O	O
both	NN	O	O
.	NN	O	O

-DOCSTART-	O

Carboxyl-terminal	NN	O	B-protein
15-amino	NN	O	I-protein
acid	NN	O	I-protein
sequence	NN	O	I-protein
of	NN	O	O
NFATx1	NN	O	B-protein
is	NN	O	O
possibly	NN	O	O
created	NN	O	O
by	NN	O	O
tissue-specific	NN	O	O
splicing	NN	O	O
and	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
transactivation	NN	O	O
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NFAT	NN	O	B-protein
regulates	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
cytokine	NN	O	O
and	NN	O	O
other	NN	O	O
immunoregulatory	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
isolated	NN	O	O
NFATx	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
four	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
is	NN	O	O
preferentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
an	NN	O	O
isoform	NN	O	O
of	NN	O	O
NFATx	NN	O	B-protein
,	NN	O	O
NFATx1	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
provide	NN	O	O
evidence	NN	O	O
showing	NN	O	O
that	NN	O	O
15	NN	O	O
amino	NN	O	O
acids	NN	O	O
in	NN	O	O
the	NN	O	O
carboxyl-terminal	NN	O	B-protein
end	NN	O	I-protein
of	NN	O	O
NFATx1	NN	O	B-protein
are	NN	O	O
required	NN	O	O
for	NN	O	O
its	NN	O	O
maximum	NN	O	O
transactivation	NN	O	O
activity	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
fusion	NN	O	O
between	NN	O	O
these	NN	O	O
15	NN	O	O
amino	NN	O	O
acids	NN	O	O
and	NN	O	O
the	NN	O	O
GAL4	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
was	NN	O	O
capable	NN	O	O
of	NN	O	O
transactivating	NN	O	O
reporters	NN	O	O
driven	NN	O	O
by	NN	O	O
the	NN	O	O
GAL4	NN	O	B-DNA
DNA	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
this	NN	O	O
15-amino	NN	O	B-protein
acid	NN	O	I-protein
transactivation	NN	O	I-protein
sequence	NN	O	I-protein
is	NN	O	O
well	NN	O	O
conserved	NN	O	O
in	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
although	NN	O	O
the	NN	O	O
sequences	NN	O	O
contiguous	NN	O	O
to	NN	O	O
the	NN	O	O
carboxyl-terminal	NN	O	B-protein
regions	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
family	NN	O	I-protein
are	NN	O	O
much	NN	O	O
less	NN	O	O
conserved	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
report	NN	O	O
three	NN	O	O
additional	NN	O	O
isoforms	NN	O	O
of	NN	O	O
NFATx	NN	O	B-protein
,	NN	O	O
designated	NN	O	O
NFATx2	NN	O	B-protein
,	NN	O	O
NFATx3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NFATx4	NN	O	B-protein
.	NN	O	O

This	NN	O	O
transactivation	NN	O	O
sequence	NN	O	O
is	NN	O	O
altered	NN	O	O
by	NN	O	O
tissue-specific	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
in	NN	O	O
newly	NN	O	O
isolated	NN	O	O
NFATx	NN	O	B-protein
isoforms	NN	O	I-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
lower	NN	O	O
transactivation	NN	O	O
activity	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

NFATx1	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
predominantly	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	O
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocyte	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
the	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
expressed	NN	O	O
primarily	NN	O	O
NFATx2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
activated	NN	O	O
strongly	NN	O	O
by	NN	O	O
NFATx1	NN	O	B-protein
but	NN	O	O
only	NN	O	O
weakly	NN	O	O
by	NN	O	O
NFATx2	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
15-amino	NN	O	B-protein
acid	NN	O	I-protein
sequence	NN	O	I-protein
of	NN	O	O
NFATx1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
major	NN	O	B-protein
transactivation	NN	O	I-protein
sequence	NN	O	I-protein
required	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
genes	NN	O	O
by	NN	O	O
NFATx1	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
possibly	NN	O	O
regulates	NN	O	O
NFAT	NN	O	B-protein
activity	NN	O	O
through	NN	O	O
tissue-specific	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
.	NN	O	O

-DOCSTART-	O

Altered	NN	O	B-protein
DNA-binding	NN	O	I-protein
specificity	NN	O	I-protein
mutants	NN	O	I-protein
of	NN	O	O
EKLF	NN	O	B-protein
and	NN	O	O
Sp1	NN	O	O
show	NN	O	O
that	NN	O	O
EKLF	NN	O	B-protein
is	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
locus	NN	O	I-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

The	NN	O	O
locus	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
cluster	NN	O	I-DNA
contains	NN	O	O
five	NN	O	O
DNase	NN	O	B-DNA
I	NN	O	I-DNA
hypersensitive	NN	O	I-DNA
sites	NN	O	I-DNA
(	NN	O	O
5'HS1-5	NN	O	B-DNA
)	NN	O	O
required	NN	O	O
for	NN	O	O
locus	NN	O	O
activation	NN	O	O
.	NN	O	O

5'HS3	NN	O	B-DNA
contains	NN	O	O
six	NN	O	O
G-rich	NN	O	O
motifs	NN	O	O
that	NN	O	O
are	NN	O	O
essential	NN	O	O
for	NN	O	O
its	NN	O	O
activity	NN	O	O
.	NN	O	O

Members	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	O
family	NN	O	O
,	NN	O	O
characterized	NN	O	O
by	NN	O	O
three	NN	O	O
zinc	NN	O	B-protein
fingers	NN	O	I-protein
highly	NN	O	O
homologous	NN	O	O
to	NN	O	O
those	NN	O	O
found	NN	O	O
in	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Sp1	NN	O	I-protein
,	NN	O	O
interact	NN	O	O
with	NN	O	O
these	NN	O	O
motifs	NN	O	O
.	NN	O	O

Because	NN	O	O
point	NN	O	O
mutagenesis	NN	O	O
can	NN	O	O
not	NN	O	O
distinguish	NN	O	O
between	NN	O	O
family	NN	O	O
members	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
which	NN	O	O
protein	NN	O	O
activates	NN	O	O
5'HS3	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
such	NN	O	O
closely	NN	O	O
related	NN	O	O
proteins	NN	O	O
can	NN	O	O
be	NN	O	O
distinguished	NN	O	O
in	NN	O	O
vivo	NN	O	O
by	NN	O	O
matching	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
5'HS3	NN	O	B-DNA
with	NN	O	O
amino	NN	O	O
acid	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
zinc	NN	O	B-protein
fingers	NN	O	I-protein
of	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
EKLF	NN	O	B-protein
.	NN	O	O

Testing	NN	O	O
their	NN	O	O
activity	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
shows	NN	O	O
that	NN	O	O
EKLF	NN	O	B-protein
is	NN	O	O
a	NN	O	O
direct	NN	O	O
activator	NN	O	O
of	NN	O	O
5'HS3	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
Rta	NN	O	I-protein
protein	NN	O	I-protein
activates	NN	O	O
lytic	NN	O	B-DNA
cycle	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
can	NN	O	O
disrupt	NN	O	O
latency	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
transition	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
from	NN	O	O
latency	NN	O	O
into	NN	O	O
the	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
two	NN	O	O
immediate-early	NN	O	B-DNA
viral	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
BZLF1	NN	O	B-DNA
and	NN	O	O
BRLF1	NN	O	B-DNA
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
ZEBRA	NN	O	O
,	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
BZLF1	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
disrupt	NN	O	O
latency	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
stimulating	NN	O	O
expression	NN	O	O
of	NN	O	O
lytic	NN	O	B-DNA
cycle	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
BRLF1	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
BRLF1	NN	O	B-protein
product	NN	O	I-protein
Rta	NN	O	I-protein
functions	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
in	NN	O	O
both	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
Rta	NN	O	B-protein
has	NN	O	O
recently	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
disrupt	NN	O	O
latency	NN	O	O
in	NN	O	O
an	NN	O	O
epithelial	NN	O	O
specific	NN	O	O
manner	NN	O	O
(	NN	O	O
S.	NN	O	O
Zalani	NN	O	O
,	NN	O	O
E.	NN	O	O
Holley-Guthrie	NN	O	O
,	NN	O	O
and	NN	O	O
S.	NN	O	O
Kenney	NN	O	O
,	NN	O	O
Proc.	NN	O	O
Natl.	NN	O	O
Acad.	NN	O	O
Sci.	NN	O	O
USA	NN	O	O
93	NN	O	O
:	NN	O	O
9194-9199	NN	O	O
,	NN	O	O
1996	NN	O	O
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
Rta	NN	O	B-protein
is	NN	O	O
also	NN	O	O
sufficient	NN	O	O
for	NN	O	O
disruption	NN	O	O
of	NN	O	O
latency	NN	O	O
in	NN	O	O
a	NN	O	O
permissive	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
HH514-16	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
transfection	NN	O	O
of	NN	O	O
Rta	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
synthesis	NN	O	O
of	NN	O	O
ZEBRA	NN	O	B-protein
,	NN	O	O
viral	NN	O	O
DNA	NN	O	O
replication	NN	O	O
,	NN	O	O
and	NN	O	O
late	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
Rta	NN	O	B-protein
by	NN	O	O
itself	NN	O	O
is	NN	O	O
less	NN	O	O
potent	NN	O	O
than	NN	O	O
ZEBRA	NN	O	B-protein
in	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
activate	NN	O	O
most	NN	O	O
early	NN	O	O
and	NN	O	O
late	NN	O	O
lytic	NN	O	B-DNA
cycle	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
light	NN	O	O
of	NN	O	O
previous	NN	O	O
work	NN	O	O
implicating	NN	O	O
ZEBRA	NN	O	B-protein
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Rta	NN	O	B-protein
,	NN	O	O
we	NN	O	O
suggest	NN	O	O
a	NN	O	O
cooperative	NN	O	O
model	NN	O	O
for	NN	O	O
EBV	NN	O	O
entry	NN	O	O
into	NN	O	O
the	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
either	NN	O	O
BZLF1	NN	O	B-DNA
or	NN	O	O
BRLF1	NN	O	B-DNA
triggers	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
immediate-early	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
together	NN	O	O
these	NN	O	O
activators	NN	O	O
act	NN	O	O
individually	NN	O	O
or	NN	O	O
in	NN	O	O
synergy	NN	O	O
on	NN	O	O
downstream	NN	O	B-DNA
targets	NN	O	I-DNA
to	NN	O	O
activate	NN	O	O
the	NN	O	O
viral	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	O
of	NN	O	O
a	NN	O	O
minimal	NN	O	B-DNA
promoter	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
NF-IL6	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
CREB/ATF	NN	O	B-protein
and	NN	O	O
SP1	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
U937	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

NF-IL6	NN	O	B-protein
is	NN	O	O
an	NN	O	O
important	NN	O	O
transcriptional	NN	O	O
regulator	NN	O	O
of	NN	O	O
genes	NN	O	O
induced	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
monocytes/macrophages	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
NF-IL6	NN	O	B-protein
is	NN	O	O
the	NN	O	O
only	NN	O	O
CCAAT/enhancer-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
C/EBP	NN	O	I-protein
)	NN	O	I-protein
family	NN	O	I-protein
member	NN	O	O
whose	NN	O	O
steady-state	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
increase	NN	O	O
upon	NN	O	O
activation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
(	NN	O	O
1	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
increased	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
NF-IL6	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
responsible	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-IL6	NN	O	B-RNA
mRNA	NN	O	I-RNA
following	NN	O	O
activation	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
104-bp	NN	O	B-DNA
minimal	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
NF-IL6	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
sufficient	NN	O	O
for	NN	O	O
basal	NN	O	O
and	NN	O	O
activation-dependent	NN	O	O
induction	NN	O	O
of	NN	O	O
transcription	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
region	NN	O	O
contains	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
cAMP	NN	O	B-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein/activation	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
CREB/ATF	NN	O	B-protein
)	NN	O	O
and	NN	O	O
Sp1	NN	O	B-protein
families	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Each	NN	O	O
site	NN	O	O
is	NN	O	O
functionally	NN	O	O
important	NN	O	O
and	NN	O	O
contributes	NN	O	O
independently	NN	O	O
to	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
NF-IL6	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

CIITA	NN	O	B-protein
B-cell-specific	NN	O	B-DNA
promoter	NN	O	I-DNA
suppression	NN	O	O
in	NN	O	O
MHC	NN	O	B-cell_line
class	NN	O	I-cell_line
II-silenced	NN	O	I-cell_line
cell	NN	O	I-cell_line
hybrids	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
various	NN	O	O
sets	NN	O	O
of	NN	O	O
somatic	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybrids	NN	O	I-cell_line
,	NN	O	O
generated	NN	O	O
by	NN	O	O
the	NN	O	O
fusion	NN	O	O
of	NN	O	O
epithelial	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
with	NN	O	O
B-lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
were	NN	O	O
analyzed	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
(	NN	O	I-protein
MHC	NN	O	I-protein
)	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
.	NN	O	O

We	NN	O	O
first	NN	O	O
demonstrate	NN	O	O
,	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
and	NN	O	I-cell_line
mouse	NN	O	I-cell_line
intraspecies	NN	O	I-cell_line
hybrids	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
coordinate	NN	O	O
suppression	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
,	NN	O	O
Ii	NN	O	B-protein
(	NN	O	O
invariant	NN	O	B-protein
chain	NN	O	I-protein
)	NN	O	O
and	NN	O	O
HLA-DM	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
the	NN	O	O
silencing	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
interspecies	NN	O	O
hybrids	NN	O	O
,	NN	O	O
the	NN	O	O
segregation	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
chromosomes	NN	O	I-DNA
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
establish	NN	O	O
that	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
extinction	NN	O	O
is	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
presence	NN	O	O
in	NN	O	O
the	NN	O	O
hybrids	NN	O	O
of	NN	O	O
the	NN	O	O
chromosomes	NN	O	B-DNA
from	NN	O	O
the	NN	O	O
epithelial	NN	O	B-cell_type
fusion	NN	O	I-cell_type
partner	NN	O	I-cell_type
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
MHC	NN	O	B-RNA
class	NN	O	I-RNA
II	NN	O	I-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
CIITA	NN	O	B-protein
is	NN	O	O
the	NN	O	O
actual	NN	O	O
target	NN	O	O
of	NN	O	O
the	NN	O	O
silencing	NN	O	O
.	NN	O	O

To	NN	O	O
gain	NN	O	O
further	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
suppression	NN	O	O
phenomenon	NN	O	O
we	NN	O	O
performed	NN	O	O
luciferase	NN	O	O
assays	NN	O	O
which	NN	O	O
show	NN	O	O
that	NN	O	O
silencing	NN	O	O
affects	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	B-DNA
promoter	NN	O	I-DNA
of	NN	O	O
CIITA	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
therefore	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
gene	NN	O	O
silencing	NN	O	O
in	NN	O	O
somatic	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybrids	NN	O	I-cell_line
is	NN	O	O
due	NN	O	O
to	NN	O	O
an	NN	O	O
active	NN	O	O
suppression	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
CIITA	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
epithelial	NN	O	B-cell_type
cell	NN	O	I-cell_type
fusion	NN	O	I-cell_type
partner	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Class	NN	O	O
II	NN	O	O
transactivator-independent	NN	O	O
endothelial	NN	O	O
cell	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
gene	NN	O	O
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
lymphocyte	NN	O	O
adhesion	NN	O	O
.	NN	O	O

NK	NN	O	O
cells	NN	O	O
induce	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
allogeneic	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
in	NN	O	O
an	NN	O	O
adhesion-dependent	NN	O	O
,	NN	O	O
IFN-gamma	NN	O	B-protein
-independent	NN	O	O
manner	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
NK	NN	O	B-cell_line
cells	NN	O	I-cell_line
induce	NN	O	O
HLA-DR	NN	O	B-protein
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
a	NN	O	O
mutant	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
that	NN	O	O
is	NN	O	O
defective	NN	O	O
in	NN	O	O
IFN-gamma	NN	O	B-protein
-induced	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

RNA	NN	O	O
analysis	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
cell	NN	O	O
line	NN	O	O
that	NN	O	O
is	NN	O	O
defective	NN	O	O
in	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
NK	NN	O	O
cell-induced	NN	O	O
HLA-DR	NN	O	B-RNA
alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
is	NN	O	O
also	NN	O	O
CIITA	NN	O	B-protein
-independent	NN	O	O
.	NN	O	O

The	NN	O	O
Janus	NN	O	B-cell_line
kinase-1-deficient	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U4A	NN	O	I-cell_line
expresses	NN	O	O
HLA-DR	NN	O	B-RNA
alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
NK	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
HLA-DR	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
transfected	NN	O	O
into	NN	O	O
these	NN	O	O
cells	NN	O	O
are	NN	O	O
induced	NN	O	O
by	NN	O	O
NK	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
-independent	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
target	NN	O	O
cell	NN	O	O
HLA-DR	NN	O	B-protein
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
lymphocyte	NN	O	O
adhesion	NN	O	O
uses	NN	O	O
a	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
that	NN	O	O
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
-dependent	NN	O	O
mechanism	NN	O	O
and	NN	O	O
also	NN	O	O
suggest	NN	O	O
that	NN	O	O
CIITA	NN	O	B-protein
is	NN	O	O
not	NN	O	O
required	NN	O	O
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
overexpression	NN	O	O
of	NN	O	O
IL-1	NN	O	B-protein
receptor-associated	NN	O	I-protein
kinase	NN	O	I-protein
on	NN	O	O
NFkappaB	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
and	NN	O	O
stress-activated	NN	O	O
protein	NN	O	O
kinases	NN	O	O
in	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
EL4	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
association	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1	NN	O	B-protein
receptor-associated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
IRAK	NN	O	B-protein
)	NN	O	O
to	NN	O	O
the	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
earliest	NN	O	O
events	NN	O	O
detectable	NN	O	O
in	NN	O	O
IL-1	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

We	NN	O	O
generated	NN	O	O
permanent	NN	O	O
clones	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
EL4	NN	O	I-cell_line
6.1	NN	O	I-cell_line
overexpressing	NN	O	O
human	NN	O	B-protein
(	NN	O	I-protein
h	NN	O	I-protein
)	NN	O	I-protein
IRAK	NN	O	I-protein
to	NN	O	O
evaluate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
kinase	NN	O	O
in	NN	O	O
IL-1	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
hIRAK	NN	O	B-protein
enhanced	NN	O	O
IL-1	NN	O	B-protein
-stimulated	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NFkappaB	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
truncated	NN	O	O
form	NN	O	O
(	NN	O	O
N-IRAK	NN	O	B-protein
)	NN	O	O
specifically	NN	O	O
inhibited	NN	O	O
IL-1	NN	O	B-protein
-dependent	NN	O	O
NFkappaB	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

In	NN	O	O
clones	NN	O	O
stably	NN	O	O
overexpressing	NN	O	O
hIRAK	NN	O	B-protein
a	NN	O	O
weak	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
low	NN	O	O
basal	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
which	NN	O	O
was	NN	O	O
enhanced	NN	O	O
in	NN	O	O
an	NN	O	O
IL-1	NN	O	B-protein
-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Compared	NN	O	O
to	NN	O	O
the	NN	O	O
parental	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
the	NN	O	O
dose-response	NN	O	O
curve	NN	O	O
of	NN	O	O
IL-1	NN	O	B-protein
-induced	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
was	NN	O	O
shifted	NN	O	O
in	NN	O	O
both	NN	O	O
potency	NN	O	O
and	NN	O	O
efficacy	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
IRAK	NN	O	B-protein
directly	NN	O	O
triggers	NN	O	O
NFkappaB	NN	O	B-protein
-mediated	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
EL4	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Qualitatively	NN	O	O
different	NN	O	O
effects	NN	O	O
were	NN	O	O
observed	NN	O	O
for	NN	O	O
the	NN	O	O
IL-1	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
stress-activated	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
SAP	NN	O	I-protein
)	NN	O	I-protein
kinases	NN	O	I-protein
:	NN	O	O
permanent	NN	O	O
overexpression	NN	O	O
of	NN	O	O
IRAK	NN	O	B-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
dose	NN	O	O
dependence	NN	O	O
but	NN	O	O
prolonged	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
IL-1	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
SAP	NN	O	B-protein
kinases	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
signaling	NN	O	O
branch	NN	O	O
may	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
distinct	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
signaling	NN	O	O
in	NN	O	O
activated	NN	O	O
DRA	NN	O	B-DNA
transcription	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
MHC	NN	O	I-DNA
HLA-DRA	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
can	NN	O	O
be	NN	O	O
up-regulated	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
Ig	NN	O	O
cross-linking	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
such	NN	O	O
as	NN	O	O
12-O-tetradecanoyl	NN	O	O
phorbol	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
.	NN	O	O

Induced	NN	O	O
DRA	NN	O	B-DNA
expression	NN	O	O
involves	NN	O	O
activation	NN	O	O
of	NN	O	O
restricted	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	I-protein
PKC	NN	O	I-protein
)	NN	O	I-protein
isoforms	NN	O	I-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
activated	NN	O	O
activator	NN	O	O
protein-1	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
expression	NN	O	O
profiles	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
were	NN	O	O
analyzed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
Raji	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphoblastoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Transient	NN	O	O
transfection	NN	O	O
analysis	NN	O	O
with	NN	O	O
target	NN	O	B-DNA
plasmids	NN	O	I-DNA
containing	NN	O	O
either	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
wild-type	NN	O	O
or	NN	O	O
mutated	NN	O	O
)	NN	O	O
or	NN	O	O
TPA	NN	O	B-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
the	NN	O	O
selective	NN	O	O
PKC	NN	O	B-protein
inhibitor	NN	O	O
GF	NN	O	O
109203X	NN	O	O
repressed	NN	O	O
TPA-mediated	NN	O	O
activation	NN	O	O
.	NN	O	O

Western	NN	O	O
analysis	NN	O	O
performed	NN	O	O
on	NN	O	O
cellular	NN	O	O
fractions	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
of	NN	O	O
TPA-activated	NN	O	B-cell_type
cells	NN	O	I-cell_type
revealed	NN	O	O
abundant	NN	O	O
expression	NN	O	O
of	NN	O	O
classical	NN	O	B-protein
PKC-alpha	NN	O	I-protein
(	NN	O	O
cPKC-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
cPKC-betaII	NN	O	B-protein
,	NN	O	O
and	NN	O	O
atypical	NN	O	B-protein
PKC-zeta	NN	O	I-protein
isoforms	NN	O	I-protein
and	NN	O	O
identified	NN	O	O
a	NN	O	O
sustained	NN	O	O
translocation	NN	O	O
of	NN	O	O
cPKC-alpha	NN	O	B-protein
and	NN	O	O
cPKC-betaII	NN	O	B-protein
from	NN	O	O
the	NN	O	O
cytosolic	NN	O	O
compartment	NN	O	O
to	NN	O	O
membranes	NN	O	O
.	NN	O	O

As	NN	O	O
expected	NN	O	O
,	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
atypical	NN	O	B-protein
PKC-zeta	NN	O	I-protein
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
TPA	NN	O	O
treatment	NN	O	O
and	NN	O	O
displayed	NN	O	O
an	NN	O	O
even	NN	O	O
distribution	NN	O	O
between	NN	O	O
cytosol	NN	O	O
and	NN	O	O
membranes	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
immunofluorescence	NN	O	O
microscopy	NN	O	O
.	NN	O	O

The	NN	O	O
TPA-mediated	NN	O	O
translocation	NN	O	O
of	NN	O	O
cPKC-alpha	NN	O	B-protein
and	NN	O	O
cPKC-betaII	NN	O	B-protein
was	NN	O	O
not	NN	O	O
influenced	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
GF	NN	O	O
109203X	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
functional	NN	O	O
activation	NN	O	O
and	NN	O	O
translocation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
were	NN	O	O
investigated	NN	O	O
with	NN	O	O
a	NN	O	O
selective	NN	O	O
in	NN	O	O
vitro	NN	O	O
kinase	NN	O	O
assay	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
activated	NN	O	O
HLA-DRA	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TPA	NN	O	O
treatment	NN	O	O
is	NN	O	O
strictly	NN	O	O
dependent	NN	O	O
on	NN	O	O
PKC	NN	O	B-protein
activation	NN	O	O
acting	NN	O	O
on	NN	O	O
the	NN	O	O
X2	NN	O	B-DNA
box	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
selective	NN	O	O
inhibition	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
enzymatic	NN	O	O
activity	NN	O	O
does	NN	O	O
not	NN	O	O
influence	NN	O	O
subcellular	NN	O	O
localization	NN	O	O
of	NN	O	O
expressed	NN	O	O
PKC	NN	O	B-protein
isoenzymes	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
translocation	NN	O	O
event	NN	O	O
per	NN	O	O
se	NN	O	O
occurs	NN	O	O
independently	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
RNA	NN	O	B-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
transcription	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
synthetic	NN	O	O
DNA-binding	NN	O	O
ligands	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Sequence-specific	NN	O	O
DNA-binding	NN	O	O
small	NN	O	O
molecules	NN	O	O
that	NN	O	O
can	NN	O	O
permeate	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
potentially	NN	O	O
could	NN	O	O
regulate	NN	O	O
transcription	NN	O	O
of	NN	O	O
specific	NN	O	O
genes	NN	O	O
.	NN	O	O

Multiple	NN	O	O
cellular	NN	O	B-protein
DNA-binding	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
are	NN	O	O
required	NN	O	O
by	NN	O	O
HIV	NN	O	O
type	NN	O	O
1	NN	O	O
for	NN	O	O
RNA	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Two	NN	O	O
pyrrole-imidazole	NN	O	O
polyamides	NN	O	O
were	NN	O	O
designed	NN	O	O
to	NN	O	O
bind	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
immediately	NN	O	O
adjacent	NN	O	O
to	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
Ets-1	NN	O	B-protein
,	NN	O	O
lymphoid-enhancer	NN	O	B-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
TATA-box	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

These	NN	O	O
synthetic	NN	O	O
ligands	NN	O	O
specifically	NN	O	O
inhibit	NN	O	O
DNA-binding	NN	O	O
of	NN	O	O
each	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
and	NN	O	O
HIV	NN	O	O
type	NN	O	O
1	NN	O	O
transcription	NN	O	O
in	NN	O	O
cell-free	NN	O	O
assays	NN	O	O
.	NN	O	O

When	NN	O	O
used	NN	O	O
in	NN	O	O
combination	NN	O	O
,	NN	O	O
the	NN	O	O
polyamides	NN	O	O
inhibit	NN	O	O
virus	NN	O	O
replication	NN	O	O
by	NN	O	O
>	NN	O	O
99	NN	O	O
%	NN	O	O
in	NN	O	O
isolated	NN	O	B-cell_type
human	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
with	NN	O	O
no	NN	O	O
detectable	NN	O	O
cell	NN	O	O
toxicity	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
small	NN	O	O
molecules	NN	O	O
to	NN	O	O
target	NN	O	O
predetermined	NN	O	O
DNA	NN	O	B-DNA
sequences	NN	O	I-DNA
located	NN	O	O
within	NN	O	O
RNA	NN	O	B-DNA
polymerase	NN	O	I-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
suggests	NN	O	O
a	NN	O	O
general	NN	O	O
approach	NN	O	O
for	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
viral	NN	O	O
replication	NN	O	O
.	NN	O	O

-DOCSTART-	O

musculin	NN	O	B-DNA
:	NN	O	O
a	NN	O	O
murine	NN	O	B-DNA
basic	NN	O	I-DNA
helix-loop-helix	NN	O	I-DNA
transcription	NN	O	I-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
expressed	NN	O	O
in	NN	O	O
embryonic	NN	O	O
skeletal	NN	O	O
muscle	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
the	NN	O	O
embryonic	NN	O	O
expression	NN	O	O
of	NN	O	O
musculin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
new	NN	O	O
murine	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
bHLH	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Musculin	NN	O	O
protein	NN	O	O
is	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
human	NN	O	B-protein
ABF-1	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
activated	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
to	NN	O	O
epicardin/capsulin/Pod-1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
branchial	NN	O	B-cell_type
myoblasts	NN	O	I-cell_type
,	NN	O	O
visceral	NN	O	O
and	NN	O	O
urogenital	NN	O	O
mesoderm	NN	O	O
and	NN	O	O
epicardium	NN	O	O
.	NN	O	O

In	NN	O	O
situ	NN	O	O
hybridisation	NN	O	O
revealed	NN	O	O
musculin	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
embryos	NN	O	O
was	NN	O	O
largely	NN	O	O
restricted	NN	O	O
to	NN	O	O
the	NN	O	O
embryonic	NN	O	B-cell_type
skeletal	NN	O	I-cell_type
muscle	NN	O	I-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

While	NN	O	O
all	NN	O	O
skeletal	NN	O	O
muscles	NN	O	O
expressed	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
only	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
myocytes	NN	O	O
within	NN	O	O
each	NN	O	O
muscle	NN	O	O
were	NN	O	O
positive	NN	O	O
,	NN	O	O
indicating	NN	O	O
molecular	NN	O	O
heterogeneity	NN	O	O
within	NN	O	O
fetal	NN	O	O
muscle	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Elsevier	NN	O	O
Science	NN	O	O
Ireland	NN	O	O
Ltd	NN	O	O
.	NN	O	O

All	NN	O	O
Rights	NN	O	O
Reserved	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
anergy	NN	O	O
by	NN	O	O
high	NN	O	O
concentrations	NN	O	O
of	NN	O	O
immunodominant	NN	O	O
native	NN	O	O
peptide	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
production	NN	O	O
and	NN	O	O
a	NN	O	O
block	NN	O	O
in	NN	O	O
JNK	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
anergy	NN	O	O
in	NN	O	O
antigen-specific	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
potential	NN	O	O
therapeutic	NN	O	O
value	NN	O	O
for	NN	O	O
altering	NN	O	O
pathologic	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
was	NN	O	O
undertaken	NN	O	O
to	NN	O	O
further	NN	O	O
analyze	NN	O	O
changes	NN	O	O
in	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
and	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
during	NN	O	O
anergy	NN	O	O
induction	NN	O	O
using	NN	O	O
high	NN	O	O
concentrations	NN	O	O
of	NN	O	O
native	NN	O	O
peptide	NN	O	O
ligand	NN	O	O
of	NN	O	O
tetanus	NN	O	B-cell_line
toxoid	NN	O	I-cell_line
(	NN	O	I-cell_line
TT	NN	O	I-cell_line
)	NN	O	I-cell_line
-	NN	O	I-cell_line
and	NN	O	I-cell_line
myelin	NN	O	I-cell_line
basic	NN	O	I-cell_line
protein	NN	O	I-cell_line
(	NN	O	I-cell_line
MBP	NN	O	I-cell_line
)	NN	O	I-cell_line
-specific	NN	O	I-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
TT-selected	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
could	NN	O	O
be	NN	O	O
rendered	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
its	NN	O	O
dominant	NN	O	O
epitope	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
(	NN	O	O
IC50	NN	O	O
=	NN	O	O
0.03	NN	O	O
microg/ml	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
TT-selected	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
three	NN	O	O
T	NN	O	O
cell	NN	O	O
clones	NN	O	O
established	NN	O	O
from	NN	O	O
this	NN	O	O
line	NN	O	O
,	NN	O	O
continued	NN	O	O
to	NN	O	O
produce	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
significantly	NN	O	O
increased	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
production	NN	O	O
when	NN	O	O
anergy	NN	O	O
was	NN	O	O
induced	NN	O	O
with	NN	O	O
high	NN	O	O
concentrations	NN	O	O
of	NN	O	O
the	NN	O	O
immunodominant	NN	O	O
epitope	NN	O	O
.	NN	O	O

JNK	NN	O	B-protein
enzymatic	NN	O	O
activity	NN	O	O
was	NN	O	O
blocked	NN	O	O
in	NN	O	O
anergized	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
MBP	NN	O	B-protein
-selected	NN	O	O
line	NN	O	O
could	NN	O	O
likewise	NN	O	O
be	NN	O	O
rendered	NN	O	O
unresponsive	NN	O	O
by	NN	O	O
incubation	NN	O	O
with	NN	O	O
supraoptimal	NN	O	O
concentrations	NN	O	O
of	NN	O	O
immunodominant	NN	O	O
peptide	NN	O	O
and	NN	O	O
anergy	NN	O	O
induction	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
IL-10	NN	O	B-protein
release	NN	O	O
.	NN	O	O

Both	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
could	NN	O	O
be	NN	O	O
anergized	NN	O	O
by	NN	O	O
the	NN	O	O
autopresentation	NN	O	O
of	NN	O	O
native	NN	O	O
peptide	NN	O	O
since	NN	O	O
anergy	NN	O	O
was	NN	O	O
induced	NN	O	O
in	NN	O	O
cultures	NN	O	O
lacking	NN	O	O
fresh	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
cascade	NN	O	O
is	NN	O	O
blocked	NN	O	O
when	NN	O	O
anergy	NN	O	O
is	NN	O	O
induced	NN	O	O
to	NN	O	O
high	NN	O	O
concentrations	NN	O	O
of	NN	O	O
soluble	NN	O	O
peptide	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
Academic	NN	O	O
Press	NN	O	O
.	NN	O	O

-DOCSTART-	O

Upregulation	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
6	NN	O	I-protein
and	NN	O	O
granulocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
receptors	NN	O	I-protein
by	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
CCAAT	NN	O	B-protein
enhancer	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
granulopoiesis	NN	O	O
.	NN	O	O

Cytokines	NN	O	O
stimulate	NN	O	O
granulopoiesis	NN	O	O
through	NN	O	O
signaling	NN	O	O
via	NN	O	O
receptors	NN	O	O
whose	NN	O	O
expression	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
lineage-specific	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
granulocyte	NN	O	B-RNA
colony-stimulating	NN	O	I-RNA
factor	NN	O	I-RNA
(	NN	O	I-RNA
G-CSF	NN	O	I-RNA
)	NN	O	I-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
undetectable	NN	O	O
and	NN	O	O
granulocyte	NN	O	O
maturation	NN	O	O
blocked	NN	O	O
in	NN	O	O
CCAAT	NN	O	B-protein
enhancer	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
C/EBPalpha	NN	O	B-protein
)	NN	O	O
-deficient	NN	O	O
mice	NN	O	O
.	NN	O	O

This	NN	O	O
phenotype	NN	O	O
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
G-CSF	NN	O	O
receptor-/-	NN	O	O
mice	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
other	NN	O	O
genes	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
adversely	NN	O	O
affected	NN	O	O
by	NN	O	O
loss	NN	O	O
of	NN	O	O
C/EBPalpha	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
loss	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
6	NN	O	I-protein
(	NN	O	I-protein
IL-6	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
IL-6-responsive	NN	O	B-protein
colony-forming	NN	O	I-protein
units	NN	O	I-protein
(	NN	O	O
CFU-IL6	NN	O	B-protein
)	NN	O	O
in	NN	O	O
C/EBPalpha	NN	O	B-protein
-/-	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
observed	NN	O	O
failure	NN	O	O
of	NN	O	O
granulopoiesis	NN	O	O
could	NN	O	O
be	NN	O	O
rescued	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
soluble	NN	O	O
IL-6	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
IL-6	NN	O	B-protein
or	NN	O	O
by	NN	O	O
retroviral	NN	O	O
transduction	NN	O	O
of	NN	O	O
G-CSF	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
loss	NN	O	O
of	NN	O	O
both	NN	O	O
of	NN	O	O
these	NN	O	O
receptors	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
absolute	NN	O	O
block	NN	O	O
in	NN	O	O
granulocyte	NN	O	O
maturation	NN	O	O
observed	NN	O	O
in	NN	O	O
C/EBPalpha-deficient	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
these	NN	O	O
and	NN	O	O
other	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
additional	NN	O	O
C/EBPalpha	NN	O	B-DNA
target	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
possibly	NN	O	O
other	NN	O	O
cytokine	NN	O	O
receptors	NN	O	O
,	NN	O	O
are	NN	O	O
also	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
block	NN	O	O
in	NN	O	O
granulocyte	NN	O	O
differentiation	NN	O	O
observed	NN	O	O
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
C/EBPalpha	NN	O	B-protein
-deficient	NN	O	O
mice	NN	O	O
.	NN	O	O

-DOCSTART-	O

Phosphatidylinositides	NN	O	O
bind	NN	O	O
to	NN	O	O
plasma	NN	O	B-protein
membrane	NN	O	I-protein
CD14	NN	O	I-protein
and	NN	O	O
can	NN	O	O
prevent	NN	O	O
monocyte	NN	O	O
activation	NN	O	O
by	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
.	NN	O	O

Although	NN	O	O
bacterial	NN	O	O
lipopolysaccharides	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
and	NN	O	O
several	NN	O	O
other	NN	O	O
microbial	NN	O	O
agonists	NN	O	O
can	NN	O	O
bind	NN	O	O
to	NN	O	O
mCD14	NN	O	B-protein
(	NN	O	O
membrane	NN	O	B-protein
CD14	NN	O	I-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
cell-surface	NN	O	O
receptor	NN	O	O
found	NN	O	O
principally	NN	O	O
on	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
host-derived	NN	O	O
mCD14	NN	O	B-protein
ligands	NN	O	O
are	NN	O	O
poorly	NN	O	O
defined	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
phosphatidylinositol	NN	O	O
(	NN	O	O
PtdIns	NN	O	O
)	NN	O	O
,	NN	O	O
phosphatidylinositol-4-phosphate	NN	O	O
,	NN	O	O
and	NN	O	O
other	NN	O	O
phosphatidylinositides	NN	O	O
can	NN	O	O
bind	NN	O	O
to	NN	O	O
mCD14	NN	O	B-protein
.	NN	O	O

Phosphatidylserine	NN	O	O
(	NN	O	O
PS	NN	O	O
)	NN	O	O
,	NN	O	O
another	NN	O	O
anionic	NN	O	O
glycerophospholipid	NN	O	O
,	NN	O	O
binds	NN	O	O
to	NN	O	O
mCD14	NN	O	B-protein
with	NN	O	O
lower	NN	O	O
apparent	NN	O	O
affinity	NN	O	O
than	NN	O	O
does	NN	O	O
PtdIns	NN	O	O
.	NN	O	O

LPS-binding	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
a	NN	O	O
lipid	NN	O	O
transfer	NN	O	O
protein	NN	O	O
found	NN	O	O
in	NN	O	O
serum	NN	O	O
,	NN	O	O
facilitates	NN	O	O
both	NN	O	O
PS	NN	O	O
-and	NN	O	O
PtdIns-	NN	O	O
mCD14	NN	O	B-protein
binding	NN	O	O
.	NN	O	O

PtdIns	NN	O	O
binding	NN	O	O
to	NN	O	O
mCD14	NN	O	B-protein
can	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
anti-CD14	NN	O	B-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
that	NN	O	O
inhibit	NN	O	O
LPS	NN	O	O
-mCD14	NN	O	B-protein
binding	NN	O	O
,	NN	O	O
and	NN	O	O
PtdIns	NN	O	O
can	NN	O	O
inhibit	NN	O	O
both	NN	O	O
LPS	NN	O	O
-mCD14	NN	O	B-protein
binding	NN	O	O
and	NN	O	O
LPS-induced	NN	O	O
responses	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Serum-equilibrated	NN	O	B-cell_type
PtdIns	NN	O	I-cell_type
also	NN	O	I-cell_type
binds	NN	O	I-cell_type
to	NN	O	I-cell_type
mCD14-expressing	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
raising	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
endogenous	NN	O	O
PtdIns	NN	O	O
may	NN	O	O
modulate	NN	O	O
cellular	NN	O	O
responses	NN	O	O
to	NN	O	O
LPS	NN	O	O
and	NN	O	O
other	NN	O	O
mCD14	NN	O	B-protein
ligands	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
regulates	NN	O	O
Fas	NN	O	B-protein
(	NN	O	O
CD95/APO-1	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
.	NN	O	O

Fas	NN	O	B-protein
(	NN	O	O
CD95/APO-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
transmembrane	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
the	NN	O	O
TNF/neuron	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Ligation	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
by	NN	O	O
specific	NN	O	B-protein
Abs	NN	O	I-protein
or	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
FasL/CD95	NN	O	B-protein
ligand	NN	O	I-protein
)	NN	O	O
induces	NN	O	O
rapid	NN	O	O
apoptotic	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Despite	NN	O	O
progress	NN	O	O
in	NN	O	O
understanding	NN	O	O
the	NN	O	O
death	NN	O	O
signals	NN	O	O
transduced	NN	O	O
from	NN	O	O
Fas	NN	O	B-protein
,	NN	O	O
very	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
with	NN	O	O
regard	NN	O	O
to	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
Fas	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
regulated	NN	O	O
.	NN	O	O

Using	NN	O	O
our	NN	O	O
previously	NN	O	O
established	NN	O	O
murine	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
hybridoma	NN	O	I-cell_line
model	NN	O	I-cell_line
A1.1	NN	O	B-cell_line
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
specific	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
inhibitors	NN	O	O
could	NN	O	O
block	NN	O	O
activation-induced	NN	O	O
Fas	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
with	NN	O	O
PMA	NN	O	O
or	NN	O	O
1-oleoyl-2-acetyl-sn-glycerol	NN	O	O
could	NN	O	O
mimic	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
signal	NN	O	O
by	NN	O	O
inducing	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
but	NN	O	O
not	NN	O	O
FasL	NN	O	O
.	NN	O	O

PKC	NN	O	B-protein
-dependent	NN	O	O
Fas	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
several	NN	O	O
murine	NN	O	B-cell_line
and	NN	O	I-cell_line
human	NN	O	I-cell_line
tumor	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Since	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
redistribution	NN	O	O
by	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
mobilization	NN	O	O
,	NN	O	O
8-	NN	O	O
(	NN	O	O
diethylamino	NN	O	O
)	NN	O	O
-octyl-3	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
5-trimethoxybenzoate	NN	O	O
hydrochloride	NN	O	O
,	NN	O	O
inhibited	NN	O	O
TCR-induced	NN	O	B-protein
FasL	NN	O	I-protein
but	NN	O	O
not	NN	O	O
Fas	NN	O	B-protein
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
be	NN	O	O
independent	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
mobilization	NN	O	O
.	NN	O	O

Significantly	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
newly	NN	O	O
identified	NN	O	O
Fas-regulatory	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
TDAG51	NN	O	B-DNA
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
dependent	NN	O	O
upon	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
.	NN	O	O

PKC	NN	O	B-protein
activation	NN	O	O
only	NN	O	O
induced	NN	O	O
Fas	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
cells	NN	O	O
expressing	NN	O	O
wild-type	NN	O	O
TDAG51	NN	O	B-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Fas	NN	O	B-protein
expression	NN	O	O
is	NN	O	O
likely	NN	O	O
mediated	NN	O	O
by	NN	O	O
PKC	NN	O	B-protein
through	NN	O	O
TDAG51	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Retinoic	NN	O	O
acid	NN	O	O
inhibits	NN	O	O
CD40	NN	O	B-protein
+	NN	O	O
interleukin-4	NN	O	B-protein
-mediated	NN	O	O
IgE	NN	O	B-protein
production	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
in	NN	O	O
anti-CD40	NN	O	B-protein
+	NN	O	O
interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
-mediated	NN	O	O
B-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
10	NN	O	O
(	NN	O	O
-12	NN	O	O
)	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
mol/L	NN	O	O
RA	NN	O	O
was	NN	O	O
studied	NN	O	O
in	NN	O	O
anti-CD40	NN	O	B-protein
(	NN	O	O
1	NN	O	O
microgram/mL	NN	O	O
)	NN	O	O
+	NN	O	O
IL-4	NN	O	B-protein
(	NN	O	O
5	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
-mediated	NN	O	O
proliferation	NN	O	O
and	NN	O	O
Ig	NN	O	O
synthesis	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
healthy	NN	O	O
donors	NN	O	O
.	NN	O	O

Anti-CD40	NN	O	B-protein
+	NN	O	O
IL-4	NN	O	B-protein
-mediated	NN	O	O
proliferation	NN	O	O
of	NN	O	O
PBMC	NN	O	B-cell_type
and	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
RA	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
,	NN	O	O
with	NN	O	O
maximal	NN	O	O
inhibition	NN	O	O
of	NN	O	O
62	NN	O	O
%	NN	O	O
+/-	NN	O	O
5	NN	O	O
%	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
and	NN	O	O
55	NN	O	O
%	NN	O	O
+/-	NN	O	O
4.4	NN	O	O
%	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
,	NN	O	O
and	NN	O	O
58	NN	O	O
%	NN	O	O
+/-	NN	O	O
6.7	NN	O	O
%	NN	O	O
and	NN	O	O
51	NN	O	O
%	NN	O	O
+/-	NN	O	O
4.7	NN	O	O
%	NN	O	O
,	NN	O	O
respectively	NN	O	O
by	NN	O	O
13-cis	NN	O	O
RA	NN	O	O
.	NN	O	O

IgE	NN	O	B-protein
synthesis	NN	O	O
was	NN	O	O
even	NN	O	O
more	NN	O	O
markedly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
RA	NN	O	O
starting	NN	O	O
at	NN	O	O
concentrations	NN	O	O
of	NN	O	O
>	NN	O	O
10	NN	O	O
(	NN	O	O
-14	NN	O	O
)	NN	O	O
mol/L	NN	O	O
for	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
>	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
mol/L	NN	O	O
for	NN	O	O
PBMC	NN	O	B-cell_type
.	NN	O	O

Maximal	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
production	NN	O	O
for	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
at	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
mol/L	NN	O	O
for	NN	O	O
all-trans	NN	O	O
RA	NN	O	O
(	NN	O	O
94	NN	O	O
%	NN	O	O
+/-	NN	O	O
1.8	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
96	NN	O	O
%	NN	O	O
+/-	NN	O	O
3.2	NN	O	O
%	NN	O	O
for	NN	O	O
13-cis	NN	O	O
RA	NN	O	O
.	NN	O	O

Low	NN	O	O
concentrations	NN	O	O
of	NN	O	O
RA	NN	O	O
inhibiting	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
mol/L	NN	O	O
)	NN	O	O
affected	NN	O	O
neither	NN	O	O
B-cell	NN	O	O
proliferation	NN	O	O
nor	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IgA	NN	O	B-protein
,	NN	O	O
IgG	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IgM	NN	O	B-protein
.	NN	O	O

Elucidation	NN	O	O
of	NN	O	O
the	NN	O	O
mechanism	NN	O	O
involved	NN	O	O
in	NN	O	O
this	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
production	NN	O	O
shows	NN	O	O
that	NN	O	O
epsilon	NN	O	O
germline	NN	O	B-DNA
transcription	NN	O	O
is	NN	O	O
decreased	NN	O	O
by	NN	O	O
RA	NN	O	O
,	NN	O	O
whereas	NN	O	O
production	NN	O	O
of	NN	O	O
interferon-gamma	NN	O	B-protein
(	NN	O	O
IFN-gamma	NN	O	B-protein
)	NN	O	O
was	NN	O	O
not	NN	O	O
enhanced	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
RA	NN	O	O
.	NN	O	O

To	NN	O	O
differentiate	NN	O	O
whether	NN	O	O
the	NN	O	O
RA	NN	O	O
effect	NN	O	O
was	NN	O	O
mediated	NN	O	O
by	NN	O	O
RA	NN	O	B-protein
receptors	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	I-protein
beta	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
gamma	NN	O	I-protein
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
)	NN	O	O
was	NN	O	O
examined	NN	O	O
by	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT-PCR	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
unstimulated	NN	O	B-cell_type
human	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
mRNA	NN	O	O
of	NN	O	O
the	NN	O	O
RA	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	I-protein
beta	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
gamma	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
retinoids	NN	O	O
with	NN	O	O
different	NN	O	O
receptor	NN	O	O
binding	NN	O	O
specificity	NN	O	O
(	NN	O	O
CD336	NN	O	O
,	NN	O	O
CD437	NN	O	O
,	NN	O	O
CD2019	NN	O	O
,	NN	O	O
CD367	NN	O	O
)	NN	O	O
,	NN	O	O
dose-dependent	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IgE	NN	O	B-protein
synthesis	NN	O	O
was	NN	O	O
shown	NN	O	O
by	NN	O	O
all	NN	O	O
four	NN	O	O
derivates	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
most	NN	O	O
marked	NN	O	O
by	NN	O	O
an	NN	O	O
RA	NN	O	O
binding	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
receptor	NN	O	I-protein
with	NN	O	O
high	NN	O	O
specificity	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
this	NN	O	O
study	NN	O	O
shows	NN	O	O
that	NN	O	O
RA	NN	O	O
inhibits	NN	O	O
IgE	NN	O	B-protein
production	NN	O	O
of	NN	O	O
anti-CD40	NN	O	B-cell_line
+	NN	O	I-cell_line
IL-4-stimulated	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
by	NN	O	O
The	NN	O	O
American	NN	O	O
Society	NN	O	O
of	NN	O	O
Hematology	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
Tax	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
involves	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IkappaB	NN	O	B-protein
kinase	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
IKKalpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IKKbeta	NN	O	B-protein
cellular	NN	O	B-protein
kinases	NN	O	I-protein
.	NN	O	O

Tax	NN	O	B-protein
corresponds	NN	O	O
to	NN	O	O
a	NN	O	O
40-kDa	NN	O	B-protein
transforming	NN	O	I-protein
protein	NN	O	I-protein
from	NN	O	O
the	NN	O	O
pathogenic	NN	O	O
retrovirus	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
that	NN	O	O
activates	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
by	NN	O	O
an	NN	O	O
unknown	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Tax	NN	O	B-protein
expression	NN	O	O
promotes	NN	O	O
N-terminal	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
a	NN	O	O
principal	NN	O	O
cytoplasmic	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
now	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
HTLV-1	NN	O	O
Tax	NN	O	B-protein
activates	NN	O	O
the	NN	O	O
recently	NN	O	O
identified	NN	O	O
cellular	NN	O	B-protein
kinases	NN	O	I-protein
IkappaB	NN	O	B-protein
kinase	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
IKKalpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
IKKbeta	NN	O	B-protein
,	NN	O	O
which	NN	O	O
normally	NN	O	O
phosphorylate	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
on	NN	O	O
both	NN	O	O
of	NN	O	O
its	NN	O	O
N-terminal	NN	O	O
regulatory	NN	O	O
serines	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
mutant	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
termed	NN	O	O
M22	NN	O	B-protein
,	NN	O	O
which	NN	O	O
does	NN	O	O
not	NN	O	O
induce	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
fails	NN	O	O
to	NN	O	O
activate	NN	O	O
either	NN	O	O
IKKalpha	NN	O	B-protein
or	NN	O	O
IKKbeta	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
endogenous	NN	O	O
IKK	NN	O	O
enzymatic	NN	O	O
activity	NN	O	O
was	NN	O	O
significantly	NN	O	O
elevated	NN	O	O
in	NN	O	O
HTLV-1-infected	NN	O	B-cell_line
and	NN	O	I-cell_line
Tax-expressing	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
kinase-deficient	NN	O	O
mutants	NN	O	O
of	NN	O	O
IKKalpha	NN	O	B-protein
and	NN	O	O
IKKbeta	NN	O	B-protein
into	NN	O	O
either	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
or	NN	O	I-cell_line
293	NN	O	I-cell_line
cells	NN	O	I-cell_line
also	NN	O	O
inhibits	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
a	NN	O	O
kinase-deficient	NN	O	B-protein
mutant	NN	O	I-protein
of	NN	O	O
NIK	NN	O	B-protein
(	NN	O	O
NF-kappaB-inducing	NN	O	B-protein
kinase	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
represents	NN	O	O
an	NN	O	O
upstream	NN	O	O
kinase	NN	O	O
in	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	O
and	NN	O	O
IL-1	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
leading	NN	O	O
to	NN	O	O
IKKalpha	NN	O	B-protein
and	NN	O	O
IKKbeta	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
blocks	NN	O	O
Tax	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
plasma	NN	O	B-DNA
membrane-proximal	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
these	NN	O	O
proinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
pathways	NN	O	O
are	NN	O	O
apparently	NN	O	O
not	NN	O	O
involved	NN	O	O
since	NN	O	O
dominant	NN	O	O
negative	NN	O	O
mutants	NN	O	O
of	NN	O	O
the	NN	O	O
TRAF2	NN	O	B-protein
and	NN	O	I-protein
TRAF6	NN	O	I-protein
adaptors	NN	O	I-protein
,	NN	O	O
which	NN	O	O
effectively	NN	O	O
block	NN	O	O
signaling	NN	O	O
through	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
tails	NN	O	I-protein
of	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
IL-1	NN	O	B-protein
receptors	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
do	NN	O	O
not	NN	O	O
inhibit	NN	O	O
Tax	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
HTLV-1	NN	O	O
Tax	NN	O	B-protein
exploits	NN	O	O
a	NN	O	O
distal	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
proinflammatory	NN	O	B-protein
cytokine	NN	O	I-protein
signaling	NN	O	O
cascade	NN	O	O
leading	NN	O	O
to	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
pathological	NN	O	O
alteration	NN	O	O
of	NN	O	O
this	NN	O	O
cytokine	NN	O	O
pathway	NN	O	O
leading	NN	O	O
to	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
Tax	NN	O	B-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
HTLV-1-mediated	NN	O	O
transformation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
clinically	NN	O	O
manifested	NN	O	O
as	NN	O	O
the	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O

-DOCSTART-	O

Role	NN	O	O
of	NN	O	O
IKK1	NN	O	B-protein
and	NN	O	O
IKK2	NN	O	B-protein
in	NN	O	O
lipopolysaccharide	NN	O	O
signaling	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
play	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

Bacterial	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
induces	NN	O	O
monocytes	NN	O	B-cell_type
to	NN	O	O
express	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
genes	NN	O	O
by	NN	O	O
activating	NN	O	O
the	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
reported	NN	O	O
that	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
interleukin	NN	O	B-protein
1	NN	O	I-protein
signaling	NN	O	O
pathways	NN	O	O
activate	NN	O	O
two	NN	O	O
kinases	NN	O	O
,	NN	O	O
IKK1	NN	O	B-protein
and	NN	O	O
IKK2	NN	O	B-protein
.	NN	O	O

Phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
IkappaB	NN	O	B-protein
cytoplasmic	NN	O	I-protein
inhibitors	NN	O	I-protein
,	NN	O	O
IkappaBalpha	NN	O	B-protein
,	NN	O	O
IkappaBbeta	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IkappaBepsilon	NN	O	B-protein
,	NN	O	O
by	NN	O	O
these	NN	O	O
kinases	NN	O	O
triggers	NN	O	O
proteolytic	NN	O	O
degradation	NN	O	O
and	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
NF-kappaB/Rel	NN	O	B-protein
proteins	NN	O	I-protein
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

At	NN	O	O
present	NN	O	O
,	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
IKKs	NN	O	O
in	NN	O	O
LPS	NN	O	O
signaling	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
investigated	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
LPS	NN	O	O
induces	NN	O	O
IKK	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
THP-1	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
kinetics	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
kinase	NN	O	O
activity	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
relatively	NN	O	O
slow	NN	O	O
with	NN	O	O
maximal	NN	O	O
activity	NN	O	O
observed	NN	O	O
at	NN	O	O
60	NN	O	O
min	NN	O	O
,	NN	O	O
which	NN	O	O
coincides	NN	O	O
with	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkappaBs	NN	O	B-protein
and	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

In	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
,	NN	O	O
overexpression	NN	O	O
of	NN	O	O
wild	NN	O	O
type	NN	O	O
IKK1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
dominant	NN	O	B-protein
negative	NN	O	I-protein
mutant	NN	O	I-protein
IKK1	NN	O	I-protein
(	NN	O	O
K44M	NN	O	B-protein
)	NN	O	O
,	NN	O	O
or	NN	O	O
wild	NN	O	B-protein
type	NN	O	I-protein
IKK2	NN	O	I-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
LPS-induced	NN	O	O
kappaB	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
mutant	NN	O	O
of	NN	O	O
IKK2	NN	O	B-protein
inhibited	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
kappaB	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
kappaB-dependent	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
activation	NN	O	O
of	NN	O	O
IKK2	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

IL-7	NN	O	B-protein
reconstitutes	NN	O	O
multiple	NN	O	O
aspects	NN	O	O
of	NN	O	O
v-Abl	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
early	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
selectively	NN	O	O
transformed	NN	O	O
by	NN	O	O
v-Abl	NN	O	B-protein
is	NN	O	O
currently	NN	O	O
unknown	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-7	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
JAK	NN	O	I-protein
)	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
JAK3	NN	O	B-protein
,	NN	O	O
STAT5	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT6	NN	O	B-protein
,	NN	O	O
in	NN	O	O
pre-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
transformed	NN	O	O
by	NN	O	O
v-Abl	NN	O	B-protein
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
activation	NN	O	O
of	NN	O	O
these	NN	O	O
cytokine	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
by	NN	O	O
v-Abl	NN	O	B-protein
is	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
events	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
Abelson	NN	O	O
murine	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
,	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-7	NN	O	B-protein
on	NN	O	O
pre-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
transformed	NN	O	O
with	NN	O	O
a	NN	O	O
temperature-sensitive	NN	O	B-protein
v-Abl	NN	O	I-protein
mutant	NN	O	I-protein
were	NN	O	O
examined	NN	O	O
.	NN	O	O

Whereas	NN	O	O
IL-4	NN	O	B-protein
had	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
,	NN	O	O
IL-7	NN	O	B-protein
delayed	NN	O	O
both	NN	O	O
the	NN	O	O
apoptosis	NN	O	O
and	NN	O	O
cell	NN	O	O
cycle	NN	O	O
arrest	NN	O	O
that	NN	O	O
occur	NN	O	O
upon	NN	O	O
v-Abl	NN	O	B-protein
kinase	NN	O	O
inactivation	NN	O	O
.	NN	O	O

IL-7	NN	O	B-protein
also	NN	O	O
delayed	NN	O	O
the	NN	O	O
decreases	NN	O	O
in	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
c-Myc	NN	O	B-protein
,	NN	O	O
Bcl-2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Bcl-xL	NN	O	B-protein
that	NN	O	O
occur	NN	O	O
upon	NN	O	O
loss	NN	O	O
of	NN	O	O
v-Abl	NN	O	B-protein
kinase	NN	O	O
activity	NN	O	O
.	NN	O	O

IL-7	NN	O	B-protein
did	NN	O	O
not	NN	O	O
maintain	NN	O	O
v-Abl	NN	O	B-protein
-mediated	NN	O	O
differentiation	NN	O	O
arrest	NN	O	O
of	NN	O	O
the	NN	O	O
pre-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
RAG	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
IL-7	NN	O	B-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
identify	NN	O	O
a	NN	O	O
potential	NN	O	O
role	NN	O	O
for	NN	O	O
IL-7	NN	O	B-protein
signaling	NN	O	O
pathways	NN	O	O
in	NN	O	O
transformation	NN	O	O
by	NN	O	O
v-Abl	NN	O	B-protein
while	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-7	NN	O	B-protein
signaling	NN	O	O
can	NN	O	O
not	NN	O	O
substitute	NN	O	O
for	NN	O	O
an	NN	O	O
active	NN	O	O
v-Abl	NN	O	B-protein
kinase	NN	O	O
in	NN	O	O
transformed	NN	O	B-cell_type
pre-B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
regulation	NN	O	O
of	NN	O	O
coproporphyrinogen	NN	O	B-DNA
oxidase	NN	O	I-DNA
gene	NN	O	I-DNA
between	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
nonerythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Coproporphyrinogen	NN	O	B-protein
oxidase	NN	O	I-protein
(	NN	O	O
CPO	NN	O	B-protein
)	NN	O	O
catalyzes	NN	O	O
the	NN	O	O
sixth	NN	O	O
step	NN	O	O
of	NN	O	O
the	NN	O	O
heme	NN	O	B-protein
biosynthetic	NN	O	O
pathway	NN	O	O
.	NN	O	O

To	NN	O	O
assess	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
CPO	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
mouse	NN	O	B-DNA
genomic	NN	O	I-DNA
DNA	NN	O	I-DNA
clones	NN	O	I-DNA
for	NN	O	O
CPO	NN	O	B-protein
were	NN	O	O
isolated	NN	O	O
.	NN	O	O

Structural	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
CPO	NN	O	I-DNA
gene	NN	O	I-DNA
spans	NN	O	O
approximately	NN	O	O
11	NN	O	O
kb	NN	O	O
and	NN	O	O
consists	NN	O	O
of	NN	O	O
seven	NN	O	B-DNA
exons	NN	O	I-DNA
,	NN	O	O
just	NN	O	O
like	NN	O	O
its	NN	O	O
human	NN	O	O
counterpart	NN	O	O
.	NN	O	O

Functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
by	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
indicated	NN	O	O
that	NN	O	O
synergistic	NN	O	O
action	NN	O	O
between	NN	O	O
an	NN	O	O
SP-1-like	NN	O	B-DNA
element	NN	O	I-DNA
at	NN	O	O
-21/-12	NN	O	O
,	NN	O	O
a	NN	O	O
GATA	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
-59/-54	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
novel	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
CPRE	NN	O	B-DNA
(	NN	O	O
-	NN	O	O
GGACTACAG	NN	O	B-DNA
-	NN	O	O
)	NN	O	O
at	NN	O	O
-49/-41	NN	O	O
,	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
murine	NN	O	B-cell_type
erythroleukemia	NN	O	I-cell_type
(	NN	O	I-cell_type
MEL	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
nonerythroid	NN	O	B-cell_line
NIH3T3	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
GATA	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
required	NN	O	O
.	NN	O	O

Gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
specific	NN	O	O
DNA-protein	NN	O	B-protein
complexes	NN	O	I-protein
can	NN	O	O
be	NN	O	O
formed	NN	O	O
with	NN	O	O
each	NN	O	O
element	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
cell-specific	NN	O	O
differences	NN	O	O
in	NN	O	O
factors	NN	O	O
,	NN	O	O
which	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
SP-1-like	NN	O	B-DNA
element	NN	O	I-DNA
between	NN	O	O
MEL	NN	O	B-cell_line
and	NN	O	I-cell_line
NIH3T3	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
differential	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
function	NN	O	O
of	NN	O	O
CPO	NN	O	B-protein
gene	NN	O	O
between	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
nonerythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Copyright	NN	O	O
1998	NN	O	O
by	NN	O	O
The	NN	O	O
American	NN	O	O
Society	NN	O	O
of	NN	O	O
Hematology	NN	O	O

-DOCSTART-	O

Tumor	NN	O	B-protein
suppressor	NN	O	I-protein
proteins	NN	O	I-protein
as	NN	O	O
regulators	NN	O	O
of	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
products	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-DNA
suppressor	NN	O	I-DNA
genes	NN	O	I-DNA
are	NN	O	O
considered	NN	O	O
to	NN	O	O
function	NN	O	O
as	NN	O	O
specific	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
tumor	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
communication	NN	O	O
,	NN	O	O
we	NN	O	O
present	NN	O	O
evidence	NN	O	O
to	NN	O	O
show	NN	O	O
that	NN	O	O
these	NN	O	O
proteins	NN	O	O
inhibit	NN	O	O
tumor	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
by	NN	O	O
participating	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
tumor	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
ML-1	NN	O	B-cell_line
human	NN	O	I-cell_line
myeloblastic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
used	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
proliferate	NN	O	O
when	NN	O	O
treated	NN	O	O
with	NN	O	O
insulin-like	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
I	NN	O	I-protein
and	NN	O	O
transferrin	NN	O	B-protein
but	NN	O	O
differentiate	NN	O	O
to	NN	O	O
monocytes	NN	O	B-cell_type
when	NN	O	O
exposed	NN	O	O
to	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
or	NN	O	O
transforming	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
beta1	NN	O	I-protein
,	NN	O	O
or	NN	O	O
to	NN	O	O
macrophage-like	NN	O	B-cell_type
cells	NN	O	I-cell_type
when	NN	O	O
treated	NN	O	O
with	NN	O	O
both	NN	O	O
these	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

Initiation	NN	O	O
of	NN	O	O
proliferation	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
differentiation	NN	O	O
was	NN	O	O
followed	NN	O	O
by	NN	O	O
a	NN	O	O
20-	NN	O	O
to	NN	O	O
25-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
polymerase-associated	NN	O	I-protein
processivity	NN	O	I-protein
factor	NN	O	I-protein
PCNA	NN	O	I-protein
and	NN	O	O
of	NN	O	O
the	NN	O	O
proliferation-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
E2F1	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
differentiation	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
proliferation	NN	O	O
was	NN	O	O
followed	NN	O	O
by	NN	O	O
a	NN	O	O
25-	NN	O	O
to	NN	O	O
30-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
suppressor	NN	O	I-protein
proteins	NN	O	I-protein
p53	NN	O	I-protein
(	NN	O	O
wild	NN	O	O
type	NN	O	O
)	NN	O	O
,	NN	O	O
pRb	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p130/Rb2	NN	O	B-protein
and	NN	O	O
of	NN	O	O
the	NN	O	O
p53-dependent	NN	O	B-protein
cyclin	NN	O	I-protein
kinase	NN	O	I-protein
inhibitor	NN	O	I-protein
p21/Cip1	NN	O	B-protein
.	NN	O	O

p53	NN	O	B-protein
and	NN	O	O
p21/Cip1	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
inhibit	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
PCNA	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
p130	NN	O	B-protein
and	NN	O	O
pRb	NN	O	B-protein
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
inhibit	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
E2F1	NN	O	B-protein
.	NN	O	O

As	NN	O	O
a	NN	O	O
result	NN	O	O
,	NN	O	O
G1-S-associated	NN	O	B-DNA
DNA	NN	O	I-DNA
and	NN	O	O
mRNA	NN	O	O
synthesis	NN	O	O
is	NN	O	O
inhibited	NN	O	O
,	NN	O	O
growth	NN	O	O
uncoupled	NN	O	O
from	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
maturation	NN	O	O
enabled	NN	O	O
to	NN	O	O
proceed	NN	O	O
.	NN	O	O

Where	NN	O	O
this	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-protein
suppressor	NN	O	I-protein
proteins	NN	O	I-protein
is	NN	O	O
impaired	NN	O	O
,	NN	O	O
the	NN	O	O
capacity	NN	O	O
for	NN	O	O
differentiation	NN	O	O
is	NN	O	O
lost	NN	O	O
,	NN	O	O
which	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
sustained	NN	O	O
proliferation	NN	O	O
that	NN	O	O
is	NN	O	O
characteristic	NN	O	O
of	NN	O	O
the	NN	O	O
cancer	NN	O	B-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
lymphokine	NN	O	O
activated	NN	O	O
killer	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
receiving	NN	O	O
IL-2	NN	O	B-protein
immunotherapy	NN	O	O
.	NN	O	O

Administration	NN	O	O
of	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
interleukin-2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
can	NN	O	O
result	NN	O	O
in	NN	O	O
therapeutic	NN	O	O
benefits	NN	O	O
for	NN	O	O
individuals	NN	O	O
with	NN	O	O
renal	NN	O	O
cell	NN	O	O
carcinoma	NN	O	O
and	NN	O	O
melanoma	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
an	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
families	NN	O	O
AP-1	NN	O	O
,	NN	O	O
Sp1	NN	O	O
,	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	O
STAT	NN	O	B-protein
)	NN	O	O
in	NN	O	O
cancer	NN	O	B-cell_type
patients	NN	O	I-cell_type
'	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
before	NN	O	O
and	NN	O	O
after	NN	O	O
IL-2	NN	O	B-protein
immunotherapy	NN	O	O
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
a	NN	O	O
gel-shift	NN	O	O
assay	NN	O	O
.	NN	O	O

An	NN	O	O
in	NN	O	O
vitro	NN	O	O
surrogate	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
immunotherapy	NN	O	O
is	NN	O	O
the	NN	O	O
incubation	NN	O	O
of	NN	O	O
fresh	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
from	NN	O	O
healthy	NN	O	O
individuals	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
for	NN	O	O
several	NN	O	O
days	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
lymphokine	NN	O	B-protein
-activated	NN	O	O
killer	NN	O	O
(	NN	O	O
LAK	NN	O	O
)	NN	O	O
activity	NN	O	O
in	NN	O	O
these	NN	O	O
cultures	NN	O	O
.	NN	O	O

One	NN	O	O
purpose	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
describe	NN	O	O
the	NN	O	O
profile	NN	O	O
of	NN	O	O
transcription	NN	O	O
factor	NN	O	O
activation	NN	O	O
in	NN	O	O
these	NN	O	O
different	NN	O	O
populations	NN	O	O
,	NN	O	O
and	NN	O	O
assess	NN	O	O
whether	NN	O	O
the	NN	O	O
patterns	NN	O	O
observed	NN	O	O
correlated	NN	O	O
with	NN	O	O
functional	NN	O	O
differences	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Prior	NN	O	O
to	NN	O	O
in	NN	O	O
vivo	NN	O	O
IL-2	NN	O	B-protein
administration	NN	O	O
,	NN	O	O
the	NN	O	O
typical	NN	O	O
binding	NN	O	O
pattern	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
resembled	NN	O	O
that	NN	O	O
seen	NN	O	O
in	NN	O	O
fresh	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
healthy	NN	O	O
individuals	NN	O	O
.	NN	O	O

Over	NN	O	O
a	NN	O	O
3-week	NN	O	O
course	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
therapy	NN	O	O
,	NN	O	O
in	NN	O	O
most	NN	O	O
patients	NN	O	O
the	NN	O	O
binding	NN	O	O
patterns	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
proteins	NN	O	O
changed	NN	O	O
to	NN	O	O
resemble	NN	O	O
those	NN	O	O
seen	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
activated	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
cells	NN	O	O
obtained	NN	O	O
from	NN	O	O
IL-2	NN	O	B-protein
-treated	NN	O	O
patients	NN	O	O
did	NN	O	O
not	NN	O	O
have	NN	O	O
low-level	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
binding	NN	O	I-protein
factors	NN	O	I-protein
as	NN	O	O
did	NN	O	O
LAK	NN	O	B-protein
cells	NN	O	I-protein
.	NN	O	O

When	NN	O	O
these	NN	O	O
patient	NN	O	O
cells	NN	O	O
were	NN	O	O
further	NN	O	O
stimulated	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
additional	NN	O	O
differences	NN	O	O
in	NN	O	O
STAT	NN	O	B-protein
induction	NN	O	O
patterns	NN	O	O
were	NN	O	O
noted	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
further	NN	O	O
information	NN	O	O
on	NN	O	O
the	NN	O	O
molecular	NN	O	O
events	NN	O	O
occurring	NN	O	O
in	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
generated	NN	O	O
through	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
administration	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
further	NN	O	O
document	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
not	NN	O	O
a	NN	O	O
precise	NN	O	O
congruence	NN	O	O
between	NN	O	O
PBMC	NN	O	B-cell_type
activated	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
induce	NN	O	O
monocyte	NN	O	B-protein
chemotactic	NN	O	I-protein
protein-1	NN	O	I-protein
secretion	NN	O	O
and	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
intercellular	NN	O	B-cell_type
adhesion	NN	O	I-cell_type
molecule-1	NN	O	I-cell_type
on	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
Platelet/endothelium	NN	O	O
interaction	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
inflammation	NN	O	O
and	NN	O	O
atherosclerosis	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
platelets	NN	O	O
for	NN	O	O
monocyte	NN	O	B-protein
chemotactic	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
MCP-1	NN	O	B-protein
)	NN	O	O
secretion	NN	O	O
and	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
intercellular	NN	O	B-cell_type
adhesion	NN	O	I-cell_type
molecule-1	NN	O	I-cell_type
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
on	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
assessed	NN	O	O
.	NN	O	O

METHODS	NN	O	O
AND	NN	O	O
RESULTS	NN	O	O
:	NN	O	O
Monolayers	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
nonstimulated	NN	O	O
or	NN	O	O
ADP-activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
for	NN	O	O
6	NN	O	O
hours	NN	O	O
,	NN	O	O
and	NN	O	O
secretion	NN	O	O
of	NN	O	O
MCP-1	NN	O	B-protein
and	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
were	NN	O	O
determined	NN	O	O
by	NN	O	O
ELISA	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
ADP-activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
,	NN	O	O
both	NN	O	O
MCP-1	NN	O	B-protein
secretion	NN	O	O
and	NN	O	O
ICAM-1	NN	O	B-protein
surface	NN	O	O
expression	NN	O	O
were	NN	O	O
significantly	NN	O	O
increased	NN	O	O
compared	NN	O	O
with	NN	O	O
nonstimulated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.02	NN	O	O
)	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	I-protein
factor-kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
determined	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
and	NN	O	O
kappaB	NN	O	B-protein
-dependent	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
was	NN	O	O
enhanced	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
ADP-activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
induced	NN	O	O
MCP-1	NN	O	O
and	NN	O	O
ICAM-1	NN	O	O
promoter-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

Liposomal	NN	O	O
transfection	NN	O	O
of	NN	O	O
a	NN	O	O
double-stranded	NN	O	O
kappaB	NN	O	O
phosphorothioate	NN	O	O
oligonucleotide	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
the	NN	O	O
mutated	NN	O	O
form	NN	O	O
,	NN	O	O
inhibited	NN	O	O
MCP-1	NN	O	B-protein
secretion	NN	O	O
and	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
ICAM-1	NN	O	B-protein
on	NN	O	O
activated	NN	O	O
endothelium	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
The	NN	O	O
present	NN	O	O
study	NN	O	O
indicates	NN	O	O
that	NN	O	O
activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
modulate	NN	O	O
chemotactic	NN	O	O
(	NN	O	O
MCP-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
adhesive	NN	O	O
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
properties	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
an	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Platelet-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
system	NN	O	O
might	NN	O	O
contribute	NN	O	O
to	NN	O	O
early	NN	O	O
inflammatory	NN	O	O
events	NN	O	O
in	NN	O	O
atherogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
c-cerbA/thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
regulate	NN	O	O
erythroid	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Nuclear	NN	O	B-protein
receptors	NN	O	I-protein
are	NN	O	O
important	NN	O	O
regulators	NN	O	O
of	NN	O	O
erythroid	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
impact	NN	O	O
of	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RXR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
of	NN	O	O
the	NN	O	O
c-erbA/thyroid	NN	O	B-protein
hormone	NN	O	I-protein
(	NN	O	I-protein
T3	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
c-erbA/TR	NN	O	B-protein
)	NN	O	O
on	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
culture	NN	O	O
system	NN	O	O
of	NN	O	O
stem	NN	O	B-cell_type
cell	NN	O	I-cell_type
factor-dependent	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

RXR	NN	O	B-protein
,	NN	O	O
RAR	NN	O	B-protein
,	NN	O	O
and	NN	O	O
c-erbA/TR	NN	O	B-protein
-specific	NN	O	O
ligands	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
induce	NN	O	O
erythroid-specific	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
and	NN	O	O
to	NN	O	O
accelerate	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
in	NN	O	O
culture	NN	O	O
,	NN	O	O
with	NN	O	O
T3	NN	O	O
being	NN	O	O
most	NN	O	O
effective	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
while	NN	O	O
ligand-activated	NN	O	O
c-erbA/TR	NN	O	B-protein
accelerated	NN	O	O
differentiation	NN	O	O
,	NN	O	O
unliganded	NN	O	O
c-erbA/TR	NN	O	B-protein
effectively	NN	O	O
blocked	NN	O	O
differentiation	NN	O	O
and	NN	O	O
supported	NN	O	O
sustained	NN	O	O
progenitor	NN	O	O
growth	NN	O	O
in	NN	O	O
culture	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
c-erbA/TR	NN	O	B-protein
appears	NN	O	O
to	NN	O	O
act	NN	O	O
as	NN	O	O
a	NN	O	O
binary	NN	O	O
switch	NN	O	O
affecting	NN	O	O
erythroid	NN	O	B-cell_type
cell	NN	O	I-cell_type
fate	NN	O	I-cell_type
:	NN	O	O
unliganded	NN	O	O
c-erbA/TR	NN	O	B-protein
supports	NN	O	O
growth	NN	O	O
while	NN	O	O
ligand-activated	NN	O	O
c-erbA/TR	NN	O	B-protein
induces	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
impact	NN	O	O
of	NN	O	O
RXR	NN	O	B-protein
for	NN	O	O
erythroid	NN	O	O
cell	NN	O	O
development	NN	O	O
,	NN	O	O
dominant	NN	O	O
interfering	NN	O	O
mutant	NN	O	B-protein
RXRs	NN	O	I-protein
,	NN	O	O
lacking	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
functions	NN	O	O
AF-1	NN	O	B-protein
and	NN	O	O
AF-2	NN	O	B-protein
,	NN	O	O
or	NN	O	O
AF-2	NN	O	B-protein
only	NN	O	O
,	NN	O	O
or	NN	O	O
the	NN	O	O
entire	NN	O	B-DNA
DNA-binding	NN	O	I-DNA
domain	NN	O	I-DNA
,	NN	O	O
were	NN	O	O
introduced	NN	O	O
into	NN	O	O
erythroid	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
recombinant	NN	O	O
retrovirus	NN	O	O
vectors	NN	O	O
and	NN	O	O
analyzed	NN	O	O
for	NN	O	O
RXR	NN	O	B-protein
-specific	NN	O	O
effects	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
wild-type	NN	O	B-protein
RXR	NN	O	I-protein
and	NN	O	O
of	NN	O	O
the	NN	O	O
RXR	NN	O	B-protein
mutants	NN	O	I-protein
devoid	NN	O	O
of	NN	O	O
AF-1	NN	O	B-protein
and/or	NN	O	O
AF-2	NN	O	B-protein
supported	NN	O	O
a	NN	O	O
transient	NN	O	O
outgrowth	NN	O	O
of	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
marked	NN	O	O
contrast	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
dominant	NN	O	B-protein
interfering	NN	O	I-protein
deltaDNA-binding	NN	O	I-protein
domain	NN	O	I-protein
RXR	NN	O	B-protein
,	NN	O	O
containing	NN	O	O
a	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	B-DNA
DNA-binding	NN	O	I-DNA
domain	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
incompatible	NN	O	O
with	NN	O	O
erythroid	NN	O	O
cell	NN	O	O
growth	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
pivotal	NN	O	O
role	NN	O	O
of	NN	O	O
RXR	NN	O	B-protein
for	NN	O	O
erythroid	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

Peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
women	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
exhibit	NN	O	O
abnormal	NN	O	O
protein	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	B-protein
signaling	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

We	NN	O	O
examined	NN	O	O
signaling	NN	O	O
molecules	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
women	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

In	NN	O	O
6	NN	O	O
of	NN	O	O
14	NN	O	O
patients	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
displayed	NN	O	O
an	NN	O	O
impaired	NN	O	O
ability	NN	O	O
to	NN	O	O
translocate	NN	O	O
NFeB	NN	O	B-protein
p65	NN	O	I-protein
(	NN	O	O
Rel-A	NN	O	B-protein
)	NN	O	O
following	NN	O	O
activation	NN	O	O
by	NN	O	O
anti-CD3	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

This	NN	O	O
observation	NN	O	O
was	NN	O	O
made	NN	O	O
despite	NN	O	O
normal	NN	O	O
cytoplasmic	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
Rel-A	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
also	NN	O	O
detected	NN	O	O
abnormally	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
signaling	NN	O	B-protein
molecules	NN	O	I-protein
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
TCR	NN	O	I-protein
)	NN	O	I-protein
-zeta	NN	O	I-protein
,	NN	O	O
ZAP-70	NN	O	B-protein
and	NN	O	O
p56lck	NN	O	B-protein
in	NN	O	O
4	NN	O	O
of	NN	O	O
14	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
defects	NN	O	O
in	NN	O	O
T-cell	NN	O	B-protein
signaling	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
6	NN	O	O
of	NN	O	O
the	NN	O	O
14	NN	O	O
patients	NN	O	O
also	NN	O	O
exhibited	NN	O	O
an	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
dual	NN	O	O
specificity	NN	O	B-protein
phosphatase	NN	O	I-protein
,	NN	O	O
map	NN	O	B-protein
kinase	NN	O	I-protein
phosphatase-1	NN	O	I-protein
(	NN	O	O
MKP-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

MKP-1	NN	O	B-protein
inactivates	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
therefore	NN	O	O
may	NN	O	O
interfere	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
c-fos	NN	O	B-DNA
.	NN	O	O

Abnormalities	NN	O	O
of	NN	O	O
I	NN	O	O
or	NN	O	O
more	NN	O	O
signaling	NN	O	B-protein
molecules	NN	O	I-protein
were	NN	O	O
found	NN	O	O
in	NN	O	O
9	NN	O	O
of	NN	O	O
14	NN	O	O
patients	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
only	NN	O	O
3	NN	O	O
patients	NN	O	O
had	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
exhibited	NN	O	O
all	NN	O	O
5	NN	O	O
defects	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
potentially	NN	O	O
dysfunctional	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
cancer	NN	O	O
.	NN	O	O

For	NN	O	O
example	NN	O	O
,	NN	O	O
the	NN	O	O
analysis	NN	O	O
of	NN	O	O
only	NN	O	O
1	NN	O	O
signaling	NN	O	B-protein
molecule	NN	O	I-protein
may	NN	O	O
allow	NN	O	O
patients	NN	O	O
with	NN	O	O
significant	NN	O	O
defects	NN	O	O
in	NN	O	O
T-cell	NN	O	O
signaling	NN	O	O
to	NN	O	O
go	NN	O	O
unnoticed	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
despite	NN	O	O
impaired	NN	O	O
Rel-A	NN	O	B-protein
translocation	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
capable	NN	O	O
of	NN	O	O
transcribing	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Impairments	NN	O	O
in	NN	O	O
the	NN	O	O
translocation	NN	O	O
of	NN	O	O
Rel-B	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
further	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
NFKB	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
Rel-A	NN	O	B-protein
,	NN	O	O
Rel-B	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
are	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

-DOCSTART-	O

Specific	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
at	NN	O	O
different	NN	O	O
levels	NN	O	O
of	NN	O	O
human	NN	O	O
motor	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
in	NN	O	O
isolated	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
blood	NN	O	O
concentrations	NN	O	O
of	NN	O	O
hormones	NN	O	O
produced	NN	O	O
by	NN	O	O
the	NN	O	O
hypothalamic-hypophyseal-adrenocortical	NN	O	O
system	NN	O	O
in	NN	O	O
three	NN	O	O
experimental	NN	O	O
series	NN	O	O
:	NN	O	O
at	NN	O	O
normal	NN	O	O
(	NN	O	O
17	NN	O	O
subjects	NN	O	O
)	NN	O	O
,	NN	O	O
decreased	NN	O	O
(	NN	O	O
10	NN	O	O
subjects	NN	O	O
,	NN	O	O
a	NN	O	O
360-d	NN	O	O
head-down	NN	O	O
bed	NN	O	O
rest	NN	O	O
)	NN	O	O
and	NN	O	O
increased	NN	O	O
(	NN	O	O
8	NN	O	O
subjects	NN	O	O
,	NN	O	O
physical	NN	O	O
exercise	NN	O	O
on	NN	O	O
bicycle	NN	O	O
ergometer	NN	O	O
)	NN	O	O
levels	NN	O	O
of	NN	O	O
motor	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
first	NN	O	O
series	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
did	NN	O	O
not	NN	O	O
depend	NN	O	O
on	NN	O	O
the	NN	O	O
season	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
subjects	NN	O	O
nor	NN	O	O
on	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
in	NN	O	O
blood	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
second	NN	O	O
series	NN	O	O
we	NN	O	O
observed	NN	O	O
the	NN	O	O
following	NN	O	O
:	NN	O	O
at	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
first	NN	O	O
month	NN	O	O
of	NN	O	O
bed	NN	O	O
rest	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
significantly	NN	O	O
increased	NN	O	O
;	NN	O	O
at	NN	O	O
the	NN	O	O
beginning	NN	O	O
of	NN	O	O
the	NN	O	O
third	NN	O	O
month	NN	O	O
of	NN	O	O
bed	NN	O	O
rest	NN	O	O
specific	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
and	NN	O	O
circadian	NN	O	O
rhythms	NN	O	O
of	NN	O	O
adrenocorticotropin	NN	O	O
and	NN	O	O
cortisol	NN	O	O
in	NN	O	O
blood	NN	O	O
varied	NN	O	O
markedly	NN	O	O
;	NN	O	O
at	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
sixth	NN	O	O
month	NN	O	O
of	NN	O	O
bed	NN	O	O
rest	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
returned	NN	O	O
to	NN	O	O
prebed	NN	O	O
rest	NN	O	O
levels	NN	O	O
and	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
decreased	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
third	NN	O	O
series	NN	O	O
physical	NN	O	O
exercises	NN	O	O
that	NN	O	O
induced	NN	O	O
an	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
hypothalamic-hypophyseal-adrenocortical	NN	O	O
system	NN	O	O
(	NN	O	O
maximal	NN	O	O
physical	NN	O	O
exercises	NN	O	O
and	NN	O	O
prolonged	NN	O	O
submaximal	NN	O	O
exercises	NN	O	O
at	NN	O	O
70	NN	O	O
%	NN	O	O
of	NN	O	O
maximal	NN	O	O
oxygen	NN	O	O
uptake	NN	O	O
)	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
without	NN	O	O
changes	NN	O	O
of	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
both	NN	O	O
a	NN	O	O
decrease	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
of	NN	O	O
human	NN	O	O
motor	NN	O	O
activity	NN	O	O
resulted	NN	O	O
in	NN	O	O
significant	NN	O	O
changes	NN	O	O
of	NN	O	O
specific	NN	O	O
glucocorticoid	NN	O	O
binding	NN	O	O
which	NN	O	O
were	NN	O	O
not	NN	O	O
influenced	NN	O	O
by	NN	O	O
changes	NN	O	O
of	NN	O	O
circulating	NN	O	O
hormone	NN	O	O
concentrations	NN	O	O
in	NN	O	O
blood	NN	O	O
but	NN	O	O
by	NN	O	O
some	NN	O	O
other	NN	O	O
factors	NN	O	O
affected	NN	O	O
by	NN	O	O
physical	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
small	NN	O	B-protein
GTP-binding	NN	O	I-protein
protein	NN	O	I-protein
Rho	NN	O	I-protein
potentiates	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
Rho	NN	O	B-protein
family	NN	O	O
of	NN	O	O
small	NN	O	B-protein
GTP-binding	NN	O	I-protein
proteins	NN	O	I-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytoskeletal	NN	O	O
structure	NN	O	O
,	NN	O	O
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
specific	NN	O	O
cell	NN	O	O
fate	NN	O	O
development	NN	O	O
,	NN	O	O
and	NN	O	O
transformation	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
in	NN	O	O
this	NN	O	O
report	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
an	NN	O	O
activated	NN	O	O
form	NN	O	O
of	NN	O	O
Rho	NN	O	B-protein
enhances	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
activated	NN	O	O
Rho	NN	O	B-protein
(	NN	O	O
V14Rho	NN	O	B-protein
)	NN	O	O
has	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
Oct-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
activities	NN	O	O
under	NN	O	O
similar	NN	O	O
conditions	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
a	NN	O	O
V14Rho	NN	O	B-DNA
construct	NN	O	I-DNA
incapable	NN	O	O
of	NN	O	O
membrane	NN	O	O
localization	NN	O	O
(	NN	O	O
CAAX	NN	O	B-DNA
deleted	NN	O	I-DNA
)	NN	O	O
abolishes	NN	O	O
PMA-induced	NN	O	O
AP-1	NN	O	B-protein
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
Rho	NN	O	B-protein
on	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
pathway	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
dominant-negative	NN	O	B-protein
MEK	NN	O	I-protein
and	NN	O	O
a	NN	O	O
MEK	NN	O	B-protein
inhibitor	NN	O	O
(	NN	O	O
PD98059	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
Rho	NN	O	B-protein
-induced	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

V14Rho	NN	O	B-protein
binds	NN	O	O
strongly	NN	O	O
to	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
Calpha	NN	O	I-protein
(	NN	O	O
PKCalpha	NN	O	B-protein
)	NN	O	O
in	NN	O	O
vivo	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
CAAX	NN	O	B-DNA
site	NN	O	I-DNA
on	NN	O	O
V14Rho	NN	O	B-protein
severely	NN	O	O
diminished	NN	O	O
this	NN	O	O
association	NN	O	O
.	NN	O	O

Evidence	NN	O	O
for	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
PKCalpha	NN	O	B-protein
as	NN	O	O
an	NN	O	O
effector	NN	O	O
of	NN	O	O
Rho	NN	O	B-protein
was	NN	O	O
obtained	NN	O	O
by	NN	O	O
the	NN	O	O
observation	NN	O	O
that	NN	O	O
coexpression	NN	O	O
of	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
PKCalpha	NN	O	B-protein
blocked	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
activated	NN	O	O
Rho	NN	O	B-protein
plus	NN	O	O
PMA	NN	O	O
on	NN	O	O
AP-1	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
Rho	NN	O	B-protein
potentiates	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O

-DOCSTART-	O

Changes	NN	O	O
in	NN	O	O
PKC	NN	O	B-protein
isoforms	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
alveolar	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
compared	NN	O	O
with	NN	O	O
blood	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Alveolar	NN	O	O
macrophages	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
host	NN	O	O
defense	NN	O	O
and	NN	O	O
in	NN	O	O
other	NN	O	O
types	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
processes	NN	O	O
in	NN	O	O
the	NN	O	O
lung	NN	O	O
.	NN	O	O

These	NN	O	O
cells	NN	O	O
exhibit	NN	O	O
many	NN	O	O
alterations	NN	O	O
in	NN	O	O
function	NN	O	O
compared	NN	O	O
with	NN	O	O
their	NN	O	O
precursor	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
blood	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
evaluate	NN	O	O
a	NN	O	O
potential	NN	O	O
mechanism	NN	O	O
for	NN	O	O
these	NN	O	O
differences	NN	O	O
in	NN	O	O
function	NN	O	O
,	NN	O	O
we	NN	O	O
evaluated	NN	O	O
expression	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	I-protein
PKC	NN	O	I-protein
)	NN	O	I-protein
isoforms	NN	O	I-protein
.	NN	O	O

We	NN	O	O
found	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
Ca2+-dependent	NN	O	B-protein
PKC	NN	O	I-protein
isoforms	NN	O	I-protein
in	NN	O	O
monocytes	NN	O	O
compared	NN	O	O
with	NN	O	O
alveolar	NN	O	O
macrophages	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
found	NN	O	O
differential	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Ca2+-independent	NN	O	B-protein
isoforms	NN	O	I-protein
in	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
compared	NN	O	O
with	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

One	NN	O	O
consequence	NN	O	O
of	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
can	NN	O	O
be	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	I-protein
MAP	NN	O	I-protein
)	NN	O	I-protein
kinase	NN	O	I-protein
pathways	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
also	NN	O	O
evaluated	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinase	NN	O	I-protein
extracellular	NN	O	I-protein
signal-regulated	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
ERK	NN	O	I-protein
)	NN	O	I-protein
2	NN	O	I-protein
by	NN	O	O
the	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
.	NN	O	O

PMA	NN	O	O
activated	NN	O	O
ERK2	NN	O	B-protein
kinase	NN	O	I-protein
in	NN	O	O
both	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
;	NN	O	O
however	NN	O	O
,	NN	O	O
monocytes	NN	O	B-cell_type
consistently	NN	O	O
showed	NN	O	O
a	NN	O	O
significantly	NN	O	O
greater	NN	O	O
activation	NN	O	O
of	NN	O	O
ERK2	NN	O	B-protein
kinase	NN	O	I-protein
by	NN	O	O
PMA	NN	O	O
compared	NN	O	O
with	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Another	NN	O	O
known	NN	O	O
consequence	NN	O	O
of	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
and	NN	O	O
subsequent	NN	O	O
activation	NN	O	O
of	NN	O	O
ERK	NN	O	B-protein
kinase	NN	O	I-protein
is	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
activator	NN	O	I-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
evaluated	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
by	NN	O	O
PMA	NN	O	O
in	NN	O	O
both	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
very	NN	O	O
little	NN	O	O
detectable	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
in	NN	O	O
a	NN	O	O
gel	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
in	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
monocytes	NN	O	B-cell_type
showed	NN	O	O
a	NN	O	O
substantial	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
by	NN	O	O
PMA	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
differential	NN	O	O
expression	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
isoforms	NN	O	I-protein
in	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
and	NN	O	O
blood	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
important	NN	O	O
functional	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
gamma-IFN	NN	O	B-DNA
responsive	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
scavenger	NN	O	O
receptor	NN	O	O
over-expressing	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
xanthomatosis	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
described	NN	O	O
an	NN	O	O
inherited	NN	O	O
over-expression	NN	O	O
of	NN	O	O
the	NN	O	O
macrophage	NN	O	B-protein
scavenger	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
SR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
blood	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
from	NN	O	O
members	NN	O	O
of	NN	O	O
a	NN	O	O
kindred	NN	O	O
,	NN	O	O
only	NN	O	O
two	NN	O	O
of	NN	O	O
whom	NN	O	O
displayed	NN	O	O
extensive	NN	O	O
xanthomatosis	NN	O	O
.	NN	O	O

Using	NN	O	O
mRNA	NN	O	O
differential	NN	O	O
display	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
abnormally	NN	O	O
high	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	O
STAT1alpha	NN	O	B-protein
)	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
proband	NN	O	B-cell_line
II-2	NN	O	I-cell_line
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
gamma-interferon	NN	O	B-protein
inducible	NN	O	I-protein
protein	NN	O	I-protein
10	NN	O	I-protein
(	NN	O	O
IP-10	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
STAT1alpha-responsive	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
mediator	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
,	NN	O	O
was	NN	O	O
also	NN	O	O
abnormally	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
II-2	NN	O	B-cell_line
.	NN	O	O

Over-expression	NN	O	O
of	NN	O	O
both	NN	O	O
genes	NN	O	O
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
II-2	NN	O	B-cell_line
and	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
clinically	NN	O	O
unaffected	NN	O	O
family	NN	O	O
members	NN	O	O
,	NN	O	O
unlike	NN	O	O
that	NN	O	O
of	NN	O	O
SR	NN	O	B-protein
.	NN	O	O

Gel	NN	O	O
retardation	NN	O	O
assays	NN	O	O
with	NN	O	O
THP-1	NN	O	B-cell_line
cell	NN	O	I-cell_line
extracts	NN	O	O
identified	NN	O	O
gamma-IFN	NN	O	B-protein
inducible	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
to	NN	O	O
three	NN	O	O
potential	NN	O	O
STATI	NN	O	B-DNA
DNA	NN	O	I-DNA
binding	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IP-10	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
from	NN	O	O
nucleotides	NN	O	B-DNA
-	NN	O	I-DNA
245	NN	O	I-DNA
to	NN	O	I-DNA
-	NN	O	I-DNA
188	NN	O	I-DNA
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
gamma-interferon	NN	O	O
mediated	NN	O	O
cell	NN	O	O
activation	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
STAT1alpha	NN	O	B-protein
-induced	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IP-10	NN	O	B-protein
gene	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
macrophages	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
II-2	NN	O	B-cell_line
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
familial	NN	O	O
hypercholesterolemic	NN	O	O
(	NN	O	O
FH	NN	O	O
)	NN	O	O
subjects	NN	O	O
,	NN	O	O
who	NN	O	O
frequently	NN	O	O
develop	NN	O	O
xanthomatosis	NN	O	O
,	NN	O	O
revealed	NN	O	O
a	NN	O	O
significant	NN	O	O
number	NN	O	O
of	NN	O	O
subjects	NN	O	O
with	NN	O	O
elevated	NN	O	O
STAT1alpha	NN	O	O
and	NN	O	O
IP-10	NN	O	O
expression	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
effects	NN	O	O
of	NN	O	O
gamma-IFN	NN	O	B-protein
signaling	NN	O	O
could	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
foam	NN	O	O
cell	NN	O	O
formation	NN	O	O
and	NN	O	O
xanthomatosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Redox	NN	O	O
signals	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Accumulating	NN	O	O
data	NN	O	O
from	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
laboratories	NN	O	O
have	NN	O	O
recently	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	B-protein
to	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
redox	NN	O	O
homeostasis	NN	O	O
of	NN	O	O
cells	NN	O	O
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
modulating	NN	O	O
immune	NN	O	O
function	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
has	NN	O	O
been	NN	O	O
recognized	NN	O	O
to	NN	O	O
regulate	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
genes	NN	O	O
necessary	NN	O	O
for	NN	O	O
normal	NN	O	O
T	NN	O	O
cell	NN	O	O
responses	NN	O	O
including	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
,	NN	O	O
IL-8	NN	O	B-protein
,	NN	O	O
and	NN	O	O
several	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Diminished	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
occur	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
aging	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
impaired	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
might	NN	O	O
occur	NN	O	O
during	NN	O	O
cellular	NN	O	O
senescence	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
aberrancies	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
activity	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
immunopathogenesis	NN	O	O
of	NN	O	O
diseases	NN	O	O
involving	NN	O	O
immune	NN	O	O
or	NN	O	O
inflammatory	NN	O	O
processes	NN	O	O
such	NN	O	O
as	NN	O	O
atherosclerosis	NN	O	O
and	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
H2O2	NN	O	O
and	NN	O	O
other	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
as	NN	O	O
an	NN	O	O
integratory	NN	O	O
secondary	NN	O	O
messenger	NN	O	O
for	NN	O	O
divergent	NN	O	O
T	NN	O	O
cell	NN	O	O
signals	NN	O	O
has	NN	O	O
been	NN	O	O
complicated	NN	O	O
by	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
various	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
differ	NN	O	O
markedly	NN	O	O
in	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
oxidant	NN	O	O
stress	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
proposed	NN	O	O
pathways	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
have	NN	O	O
been	NN	O	O
based	NN	O	O
on	NN	O	O
indirect	NN	O	O
evidence	NN	O	O
provided	NN	O	O
by	NN	O	O
experiments	NN	O	O
which	NN	O	O
used	NN	O	O
antioxidants	NN	O	O
to	NN	O	O
inhibit	NN	O	O
active	NN	O	O
NF-kappaB	NN	O	B-protein
formation	NN	O	O
.	NN	O	O

Further	NN	O	O
,	NN	O	O
complete	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
signals	NN	O	O
,	NN	O	O
one	NN	O	O
that	NN	O	O
stimulates	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
and	NN	O	O
one	NN	O	O
that	NN	O	O
stimulates	NN	O	O
enzymatic	NN	O	O
processes	NN	O	O
including	NN	O	O
kinases	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
substantial	NN	O	O
evidence	NN	O	O
indicates	NN	O	O
that	NN	O	O
full	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
requires	NN	O	O
dual	NN	O	O
signals	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
H2O2	NN	O	O
or	NN	O	O
other	NN	O	O
ROS	NN	O	O
to	NN	O	O
induce	NN	O	O
T	NN	O	O
cell	NN	O	O
signals	NN	O	O
and	NN	O	O
functional	NN	O	O
responses	NN	O	O
by	NN	O	O
these	NN	O	O
two	NN	O	O
mechanisms	NN	O	O
is	NN	O	O
reviewed	NN	O	O
and	NN	O	O
the	NN	O	O
specific	NN	O	O
response	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
to	NN	O	O
redox	NN	O	O
changes	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
examined	NN	O	O
.	NN	O	O

Data	NN	O	O
are	NN	O	O
also	NN	O	O
presented	NN	O	O
to	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
redox	NN	O	O
regulation	NN	O	O
in	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
may	NN	O	O
be	NN	O	O
relevant	NN	O	O
to	NN	O	O
immune-related	NN	O	O
diseases	NN	O	O
and	NN	O	O
to	NN	O	O
aging	NN	O	O
.	NN	O	O

-DOCSTART-	O

Uncoupling	NN	O	O
of	NN	O	O
nonreceptor	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
from	NN	O	O
PLC-gamma1	NN	O	B-protein
in	NN	O	O
an	NN	O	O
SLP-76-deficient	NN	O	O
T	NN	O	O
cell	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
nonreceptor	NN	O	B-protein
protein	NN	O	I-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
(	NN	O	O
PTKs	NN	O	B-protein
)	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
responsiveness	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
individual	NN	O	O
PTK	NN	O	B-protein
substrates	NN	O	I-protein
is	NN	O	O
often	NN	O	O
uncertain	NN	O	O
.	NN	O	O

A	NN	O	O
mutant	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
was	NN	O	O
isolated	NN	O	O
that	NN	O	O
lacked	NN	O	O
expression	NN	O	O
of	NN	O	O
SLP-76	NN	O	B-protein
(	NN	O	O
SH2	NN	O	B-protein
domain-containing	NN	O	I-protein
leukocyte	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
76	NN	O	O
kilodaltons	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
hematopoietically	NN	O	B-protein
expressed	NN	O	I-protein
adaptor	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
PTK	NN	O	B-protein
substrate	NN	O	I-protein
.	NN	O	O

SLP-76	NN	O	B-protein
was	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
TCR	NN	O	B-protein
-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
most	NN	O	O
proteins	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
phospholipase	NN	O	B-protein
C-gamma1	NN	O	I-protein
(	NN	O	O
PLC-gamma1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
Ras	NN	O	B-protein
pathway	NN	O	O
activation	NN	O	O
.	NN	O	O

TCR	NN	O	B-protein
-inducible	NN	O	O
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
SLP-76	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
coupling	NN	O	O
of	NN	O	O
TCR-regulated	NN	O	B-protein
PTKs	NN	O	I-protein
to	NN	O	O
downstream	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
requires	NN	O	O
SLP-76	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
function	NN	O	O
of	NN	O	O
Stat1	NN	O	B-protein
and	NN	O	O
Stat3	NN	O	B-protein
proteins	NN	O	O
in	NN	O	O
erythropoietin	NN	O	B-protein
-induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
recently	NN	O	O
determined	NN	O	O
that	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	O
EPO	NN	O	B-protein
)	NN	O	O
activates	NN	O	O
3	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	O
transducer	NN	O	O
and	NN	O	O
activator	NN	O	O
of	NN	O	O
transcription	NN	O	O
(	NN	O	O
STAT	NN	O	O
)	NN	O	O
family	NN	O	O
,	NN	O	O
Stat1alpha	NN	O	B-protein
,	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Stat5	NN	O	B-protein
,	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
EPO-dependent	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
UT-7	NN	O	B-cell_line
and	NN	O	O
UT-7/EPO	NN	O	B-cell_line
(	NN	O	O
Kirito	NN	O	O
et	NN	O	O
al	NN	O	O
,	NN	O	O
J	NN	O	O
Biol	NN	O	O
Chem	NN	O	O
272	NN	O	O
:	NN	O	O
16507	NN	O	O
,	NN	O	O
1997	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
Stat1alpha	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
EPO	NN	O	B-protein
-induced	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
Stat1alpha	NN	O	B-protein
and	NN	O	O
Stat3	NN	O	B-protein
in	NN	O	O
EPO	NN	O	B-protein
-induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

UT-7/GM	NN	O	B-cell_line
was	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
model	NN	O	O
system	NN	O	O
,	NN	O	O
because	NN	O	O
this	NN	O	O
cell	NN	O	O
line	NN	O	O
can	NN	O	O
differentiate	NN	O	O
into	NN	O	O
erythroid-lineage	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
EPO	NN	O	B-protein
treatment	NN	O	O
(	NN	O	O
Komatsu	NN	O	O
et	NN	O	O
al	NN	O	O
,	NN	O	O
Blood	NN	O	O
89	NN	O	O
:	NN	O	O
4021	NN	O	O
,	NN	O	O
1997	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
EPO	NN	O	B-protein
did	NN	O	O
not	NN	O	O
activate	NN	O	O
Stat1alpha	NN	O	B-protein
or	NN	O	O
Stat3	NN	O	B-protein
in	NN	O	O
UT-7/GM	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Transfection	NN	O	O
experiments	NN	O	O
showed	NN	O	O
that	NN	O	O
both	NN	O	O
Stat1alpha	NN	O	B-protein
and	NN	O	O
Stat3	NN	O	B-protein
inhibited	NN	O	O
the	NN	O	O
induction	NN	O	O
by	NN	O	O
EPO	NN	O	B-protein
of	NN	O	O
gamma-globin	NN	O	B-RNA
and	NN	O	I-RNA
erythroid-specific	NN	O	I-RNA
5-aminolevulinate	NN	O	I-RNA
synthetase	NN	O	I-RNA
transcripts	NN	O	I-RNA
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
reduction	NN	O	O
of	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
hemoglobin-positive	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Dominant	NN	O	O
negative	NN	O	O
forms	NN	O	O
of	NN	O	O
Stat1alpha	NN	O	B-protein
or	NN	O	O
Stat3	NN	O	B-protein
promoted	NN	O	O
the	NN	O	O
EPO	NN	O	B-protein
-induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
UT-7/GM	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
although	NN	O	O
this	NN	O	O
cytokine	NN	O	O
never	NN	O	O
induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
parent	NN	O	B-cell_line
UT-7/GM	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
or	NN	O	O
without	NN	O	O
EPO	NN	O	B-protein
.	NN	O	O

A	NN	O	O
cell	NN	O	O
cycle	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
Stat1alpha	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
shortened	NN	O	O
the	NN	O	O
period	NN	O	O
of	NN	O	O
G0/G1	NN	O	O
prolongation	NN	O	O
caused	NN	O	O
by	NN	O	O
EPO	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
Stat1alpha	NN	O	B-protein
and	NN	O	O
Stat3	NN	O	B-protein
act	NN	O	O
as	NN	O	O
negative	NN	O	O
regulators	NN	O	O
in	NN	O	O
EPO	NN	O	B-protein
-induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Specifically	NN	O	O
,	NN	O	O
Stat1alpha	NN	O	B-protein
may	NN	O	O
activate	NN	O	O
a	NN	O	O
cell	NN	O	B-DNA
cycle-associated	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
s	NN	O	O
)	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
entry	NN	O	O
of	NN	O	O
cells	NN	O	O
into	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

-DOCSTART-	O

Ro	NN	O	O
09-2210	NN	O	O
exhibits	NN	O	O
potent	NN	O	O
anti-proliferative	NN	O	O
effects	NN	O	O
on	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
selectively	NN	O	O
blocking	NN	O	O
MKK	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
high	NN	O	O
throughput	NN	O	O
screening	NN	O	O
of	NN	O	O
microbial	NN	O	O
broths	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
compound	NN	O	O
,	NN	O	O
designated	NN	O	O
Ro	NN	O	O
09-2210	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
block	NN	O	O
anti-CD3	NN	O	B-protein
induced	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
with	NN	O	O
an	NN	O	O
IC50	NN	O	O
=	NN	O	O
40	NN	O	O
nM	NN	O	O
.	NN	O	O

Ro	NN	O	O
09-2210	NN	O	O
was	NN	O	O
also	NN	O	O
able	NN	O	O
to	NN	O	O
block	NN	O	O
antigen-induced	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
with	NN	O	O
an	NN	O	O
IC50	NN	O	O
=	NN	O	O
30	NN	O	O
nM	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
considerably	NN	O	O
less	NN	O	O
potent	NN	O	O
at	NN	O	O
blocking	NN	O	O
Ca2+	NN	O	O
flux	NN	O	O
stimulated	NN	O	O
by	NN	O	O
anti-CD3	NN	O	B-protein
treatment	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
Ro	NN	O	O
09-2210	NN	O	O
,	NN	O	O
we	NN	O	O
set	NN	O	O
up	NN	O	O
a	NN	O	O
transient	NN	O	O
expression	NN	O	O
system	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
using	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
reporter	NN	O	O
gene	NN	O	O
constructs	NN	O	O
and	NN	O	O
showed	NN	O	O
effective	NN	O	O
inhibition	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester/ionomycin-induced	NN	O	O
NF-AT	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
anti-CD3	NN	O	O
induced	NN	O	O
NF-AT	NN	O	B-protein
with	NN	O	O
IC50	NN	O	O
=	NN	O	O
7.7	NN	O	O
and	NN	O	O
10	NN	O	O
nM	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Ro	NN	O	O
09-2210	NN	O	O
was	NN	O	O
also	NN	O	O
able	NN	O	O
to	NN	O	O
inhibit	NN	O	O
phorbol	NN	O	O
ester/ionomycin-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
AP1	NN	O	B-protein
with	NN	O	O
IC50	NN	O	O
=	NN	O	O
<	NN	O	O
10	NN	O	O
nM	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
showed	NN	O	O
that	NN	O	O
Ro	NN	O	O
09-2210	NN	O	O
was	NN	O	O
unable	NN	O	O
to	NN	O	O
inhibit	NN	O	O
c-jun	NN	O	B-DNA
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
AP1-dependent	NN	O	B-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
(	NN	O	O
IC50	NN	O	O
>	NN	O	O
500	NN	O	O
nM	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
potently	NN	O	O
inhibit	NN	O	O
ras-induced	NN	O	O
AP1	NN	O	B-protein
activation	NN	O	O
(	NN	O	O
IC50	NN	O	O
=	NN	O	O
20	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
suggested	NN	O	O
that	NN	O	O
Ro	NN	O	O
09-2210	NN	O	O
was	NN	O	O
inhibiting	NN	O	O
an	NN	O	O
activator	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
which	NN	O	O
was	NN	O	O
upstream	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
downstream	NN	O	O
of	NN	O	O
ras	NN	O	B-protein
signaling	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
further	NN	O	O
,	NN	O	O
we	NN	O	O
then	NN	O	O
purified	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
different	NN	O	O
kinases	NN	O	O
,	NN	O	O
including	NN	O	O
PKC	NN	O	B-protein
,	NN	O	O
PhK	NN	O	B-protein
,	NN	O	O
ZAP-70	NN	O	B-protein
,	NN	O	O
ERK	NN	O	B-protein
,	NN	O	O
and	NN	O	O
MEK	NN	O	B-protein
1	NN	O	I-protein
(	NN	O	O
a	NN	O	O
MKK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
showed	NN	O	O
that	NN	O	O
Ro	NN	O	O
09-2210	NN	O	O
was	NN	O	O
a	NN	O	O
selective	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
MEK1	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
(	NN	O	O
IC50	NN	O	O
=	NN	O	O
59	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Coactivation	NN	O	O
by	NN	O	O
OCA-B	NN	O	B-protein
:	NN	O	O
definition	NN	O	O
of	NN	O	O
critical	NN	O	O
regions	NN	O	O
and	NN	O	O
synergism	NN	O	O
with	NN	O	O
general	NN	O	O
cofactors	NN	O	O
.	NN	O	O

Molecular	NN	O	O
dissection	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	O
transcription	NN	O	O
coactivator	NN	O	O
OCA-B	NN	O	B-protein
has	NN	O	O
revealed	NN	O	O
distinct	NN	O	O
regions	NN	O	O
important	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
for	NN	O	O
recruitment	NN	O	O
to	NN	O	O
immunoglobulin	NN	O	B-DNA
promoters	NN	O	I-DNA
through	NN	O	O
interaction	NN	O	O
with	NN	O	O
octamer-bound	NN	O	B-protein
Oct-1	NN	O	I-protein
and	NN	O	O
for	NN	O	O
subsequent	NN	O	O
coactivator	NN	O	O
function	NN	O	O
.	NN	O	O

Further	NN	O	O
analysis	NN	O	O
of	NN	O	O
general	NN	O	O
coactivator	NN	O	O
requirements	NN	O	O
showed	NN	O	O
that	NN	O	O
selective	NN	O	O
removal	NN	O	O
of	NN	O	O
PC4	NN	O	B-protein
from	NN	O	O
the	NN	O	O
essential	NN	O	O
USA	NN	O	O
fraction	NN	O	O
severely	NN	O	O
impairs	NN	O	O
Oct-1	NN	O	O
and	NN	O	O
OCA-B	NN	O	O
function	NN	O	O
in	NN	O	O
a	NN	O	O
cell-free	NN	O	O
system	NN	O	O
reconstituted	NN	O	O
with	NN	O	O
partially	NN	O	O
purified	NN	O	O
factors	NN	O	O
.	NN	O	O

Full	NN	O	O
activity	NN	O	O
can	NN	O	O
be	NN	O	O
restored	NN	O	O
by	NN	O	O
the	NN	O	O
combined	NN	O	O
action	NN	O	O
of	NN	O	O
recombinant	NN	O	O
PC4	NN	O	B-protein
and	NN	O	O
the	NN	O	O
PC4	NN	O	B-protein
-depleted	NN	O	O
USA	NN	O	O
fraction	NN	O	O
,	NN	O	O
thus	NN	O	O
suggesting	NN	O	O
a	NN	O	O
joint	NN	O	O
requirement	NN	O	O
for	NN	O	O
PC4	NN	O	B-protein
and	NN	O	O
another	NN	O	O
,	NN	O	O
USA-derived	NN	O	O
component	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
for	NN	O	O
optimal	NN	O	O
function	NN	O	O
of	NN	O	O
Oct-1	NN	O	B-protein
/OCA-B	NN	O	B-protein
in	NN	O	O
the	NN	O	O
reconstituted	NN	O	O
system	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
USA-derived	NN	O	B-protein
PC2	NN	O	I-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
act	NN	O	O
synergistically	NN	O	O
with	NN	O	O
PC4	NN	O	B-protein
in	NN	O	O
reproducing	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
intact	NN	O	O
USA	NN	O	O
in	NN	O	O
the	NN	O	O
assay	NN	O	O
system	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
requirement	NN	O	O
for	NN	O	O
PC4	NN	O	B-protein
in	NN	O	O
the	NN	O	O
reconstituted	NN	O	O
system	NN	O	O
,	NN	O	O
OCA-B	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
interact	NN	O	O
directly	NN	O	O
with	NN	O	O
PC4	NN	O	B-protein
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
removal	NN	O	O
of	NN	O	O
PC4	NN	O	B-protein
from	NN	O	O
the	NN	O	O
unfractionated	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
has	NN	O	O
no	NN	O	O
detrimental	NN	O	O
effect	NN	O	O
on	NN	O	O
OCA-B	NN	O	B-protein
/	NN	O	O
Oct-1	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
lead	NN	O	O
to	NN	O	O
a	NN	O	O
general	NN	O	O
model	NN	O	O
for	NN	O	O
the	NN	O	O
synergistic	NN	O	O
function	NN	O	O
of	NN	O	O
activation	NN	O	O
domains	NN	O	O
in	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
OCA-B	NN	O	B-protein
(	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
combined	NN	O	O
action	NN	O	O
of	NN	O	O
the	NN	O	O
multiple	NN	O	O
USA	NN	O	O
components	NN	O	O
)	NN	O	O
and	NN	O	O
,	NN	O	O
further	NN	O	O
,	NN	O	O
suggest	NN	O	O
a	NN	O	O
functional	NN	O	O
redundancy	NN	O	O
in	NN	O	O
general	NN	O	B-protein
coactivators	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
Stat-3	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
after	NN	O	O
ligation	NN	O	O
of	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
class	NN	O	I-protein
I	NN	O	I-protein
molecules	NN	O	I-protein
on	NN	O	O
human	NN	O	B-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
Janus	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
(	NN	O	O
Jak	NN	O	B-protein
)	NN	O	O
and	NN	O	O
Signal	NN	O	B-protein
transducers	NN	O	I-protein
and	NN	O	I-protein
activators	NN	O	I-protein
of	NN	O	I-protein
transcription	NN	O	I-protein
(	NN	O	O
Stat	NN	O	B-protein
)	NN	O	O
after	NN	O	O
ligation	NN	O	O
of	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
class	NN	O	I-protein
I	NN	O	I-protein
(	NN	O	O
MHC-I	NN	O	B-protein
)	NN	O	O
was	NN	O	O
explored	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Cross-linking	NN	O	O
of	NN	O	O
MHC-I	NN	O	B-protein
mediated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Tyk2	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Jak1	NN	O	B-protein
,	NN	O	O
Jak2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Jak3	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Stat-3	NN	O	B-protein
was	NN	O	O
tyrosine	NN	O	O
phosphorylated	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
and	NN	O	O
subsequently	NN	O	O
translocated	NN	O	O
to	NN	O	O
the	NN	O	O
cell	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Data	NN	O	O
obtained	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
activated	NN	O	B-protein
Stat-3	NN	O	I-protein
protein	NN	O	I-protein
associates	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
serum-inducible	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
hSIE	NN	O	I-DNA
)	NN	O	I-DNA
DNA-probe	NN	O	I-DNA
derived	NN	O	O
from	NN	O	O
the	NN	O	O
interferon-gamma	NN	O	B-DNA
activated	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
GAS	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
common	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
for	NN	O	O
Stat	NN	O	B-protein
protein	NN	O	I-protein
binding	NN	O	O
.	NN	O	O

An	NN	O	O
association	NN	O	O
between	NN	O	O
hSIE	NN	O	B-DNA
and	NN	O	O
Stat-3	NN	O	B-protein
after	NN	O	O
MHC-I	NN	O	B-protein
ligation	NN	O	O
was	NN	O	O
directly	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
precipitating	NN	O	O
Stat-3	NN	O	B-protein
from	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
with	NN	O	O
biotinylated	NN	O	O
hSIE	NN	O	B-DNA
probe	NN	O	I-DNA
and	NN	O	O
avidin	NN	O	B-protein
-coupled	NN	O	O
agarose	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
activated	NN	O	O
Stat-3	NN	O	B-protein
,	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
Stat-3	NN	O	B-protein
isoform	NN	O	I-protein
lacking	NN	O	O
the	NN	O	O
transactivating	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

This	NN	O	O
dominant-negative	NN	O	B-protein
acting	NN	O	I-protein
Stat-3	NN	O	I-protein
isoform	NN	O	I-protein
significantly	NN	O	O
inhibited	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
ligation	NN	O	O
of	NN	O	O
MHC-I	NN	O	B-protein
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
suggest	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
the	NN	O	O
Jak/Stat	NN	O	B-protein
signal	NN	O	O
pathway	NN	O	O
in	NN	O	O
MHC-I	NN	O	B-protein
-induced	NN	O	O
signal	NN	O	O
transduction	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

IL-2	NN	O	B-protein
-induced	NN	O	O
growth	NN	O	O
of	NN	O	O
CD8+	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
prolymphocytic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
mediated	NN	O	O
by	NN	O	O
NF-kappaB	NN	O	B-protein
induction	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
to	NN	O	O
its	NN	O	O
receptor	NN	O	O
on	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
induces	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NF-kappaB	NN	O	B-protein
)	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
IL-2R	NN	O	I-DNA
)	NN	O	I-DNA
alpha	NN	O	I-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
signaling	NN	O	O
in	NN	O	O
the	NN	O	O
growth	NN	O	O
of	NN	O	O
CD8+	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
prolymphocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
(	NN	O	I-cell_line
T-PLL	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
has	NN	O	O
been	NN	O	O
investigated	NN	O	O
.	NN	O	O

Flow	NN	O	O
cytometry	NN	O	O
revealed	NN	O	O
that	NN	O	O
primary	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
CD8+	NN	O	O
T-PLL	NN	O	O
expressed	NN	O	O
IL-2Ralpha	NN	O	B-protein
and	NN	O	I-protein
beta	NN	O	I-protein
chains	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
cells	NN	O	O
showed	NN	O	O
a	NN	O	O
proliferative	NN	O	O
response	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
IL-2Ralpha	NN	O	B-protein
expression	NN	O	O
on	NN	O	O
culture	NN	O	O
with	NN	O	O
exogeneous	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
failed	NN	O	O
to	NN	O	O
detect	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
may	NN	O	O
act	NN	O	O
in	NN	O	O
a	NN	O	O
paracrine	NN	O	O
manner	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility-shift	NN	O	O
assays	NN	O	O
revealed	NN	O	O
that	NN	O	O
recombinant	NN	O	O
IL-2	NN	O	B-protein
increased	NN	O	O
NF-kappaB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
the	NN	O	O
leukemia	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
increased	NN	O	O
the	NN	O	O
abundance	NN	O	O
of	NN	O	O
mRNAs	NN	O	O
encoding	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
components	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	O
KBF1	NN	O	B-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

IL-2	NN	O	B-protein
binding	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
markedly	NN	O	O
increased	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
low	NN	O	O
affinity	NN	O	O
IL-2Rs	NN	O	B-protein
on	NN	O	O
the	NN	O	O
leukemia	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
without	NN	O	O
an	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
high-affinity	NN	O	B-protein
IL-2Rs	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
the	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_line
CD8+	NN	O	I-cell_line
T-PLL	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
effect	NN	O	O
is	NN	O	O
mediated	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
at	NN	O	O
a	NN	O	O
pretranslational	NN	O	O
level	NN	O	O

-DOCSTART-	O

Dimethyldithiocarbamate	NN	O	O
inhibits	NN	O	O
in	NN	O	O
vitro	NN	O	O
activation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Dithiocarbamates	NN	O	O
(	NN	O	O
DTC	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
diverse	NN	O	O
group	NN	O	O
of	NN	O	O
industrial	NN	O	O
and	NN	O	O
therapeutic	NN	O	O
chemicals	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
inhibit	NN	O	O
,	NN	O	O
enhance	NN	O	O
or	NN	O	O
have	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

These	NN	O	O
apparent	NN	O	O
inconsistencies	NN	O	O
reflect	NN	O	O
the	NN	O	O
complexity	NN	O	O
of	NN	O	O
the	NN	O	O
DTCs	NN	O	O
biological	NN	O	O
activities	NN	O	O
and	NN	O	O
are	NN	O	O
probably	NN	O	O
due	NN	O	O
in	NN	O	O
part	NN	O	O
to	NN	O	O
differences	NN	O	O
in	NN	O	O
dose	NN	O	O
,	NN	O	O
route	NN	O	O
of	NN	O	O
exposure	NN	O	O
,	NN	O	O
animal	NN	O	O
species	NN	O	O
used	NN	O	O
and/or	NN	O	O
specific	NN	O	O
compound	NN	O	O
tested	NN	O	O
.	NN	O	O

The	NN	O	O
studies	NN	O	O
described	NN	O	O
herein	NN	O	O
were	NN	O	O
undertaken	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
immunotoxicity	NN	O	O
of	NN	O	O
one	NN	O	O
member	NN	O	O
of	NN	O	O
this	NN	O	O
family	NN	O	O
,	NN	O	O
dimethyldithiocarbamate	NN	O	O
(	NN	O	O
DMDTC	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
0.1-0.5	NN	O	O
microM	NN	O	O
DMDTC	NN	O	O
inhibits	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
is	NN	O	O
not	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
loss	NN	O	O
in	NN	O	O
viability	NN	O	O
,	NN	O	O
and	NN	O	O
DMDTC-treated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
retain	NN	O	O
other	NN	O	O
active	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
throughout	NN	O	O
the	NN	O	O
exposure	NN	O	O
duration	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
is	NN	O	O
apparently	NN	O	O
permanent	NN	O	O
as	NN	O	O
DMDTC-treated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
did	NN	O	O
not	NN	O	O
regain	NN	O	O
normal	NN	O	O
TNF-alpha	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
even	NN	O	O
after	NN	O	O
72	NN	O	O
h	NN	O	O
in	NN	O	O
culture	NN	O	O
.	NN	O	O

DMDTC	NN	O	O
does	NN	O	O
not	NN	O	O
appear	NN	O	O
to	NN	O	O
alter	NN	O	O
NF-kappaB	NN	O	B-protein
directly	NN	O	O
as	NN	O	O
pre-incubation	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
with	NN	O	O
DMDTC	NN	O	O
does	NN	O	O
not	NN	O	O
diminish	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
0.1-0.5	NN	O	O
microM	NN	O	O
DMDTC	NN	O	O
inhibits	NN	O	O
intracellular	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
and	NN	O	O
decreases	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
CD25	NN	O	B-protein
(	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
)	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
DMDTC	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
immunosuppressive	NN	O	O
compound	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-DNA
15-lipoxygenase	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
:	NN	O	O
analysis	NN	O	O
and	NN	O	O
identification	NN	O	O
of	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
IL-13-induced	NN	O	B-protein
regulatory	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
of	NN	O	O
15-LO	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
cloned	NN	O	O
and	NN	O	O
sequenced	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
15-LO	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
15-LO	NN	O	B-protein
promoter	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
CpG	NN	O	O
island	NN	O	O
at	NN	O	O
the	NN	O	O
5'-end	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
gene	NN	O	O
,	NN	O	O
and	NN	O	O
sequence	NN	O	O
analysis	NN	O	O
reveals	NN	O	O
putative	NN	O	O
Sp1	NN	O	B-DNA
and	NN	O	I-DNA
Ap2	NN	O	I-DNA
binding	NN	O	I-DNA
site/s	NN	O	I-DNA
and	NN	O	O
absence	NN	O	O
of	NN	O	O
TATA	NN	O	B-DNA
or	NN	O	I-DNA
CAAT	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

Transcription	NN	O	O
is	NN	O	O
initiated	NN	O	O
at	NN	O	O
one	NN	O	O
major	NN	O	O
site	NN	O	O
.	NN	O	O

Using	NN	O	O
deletion	NN	O	B-DNA
constructs	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
have	NN	O	O
defined	NN	O	O
an	NN	O	O
active	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
1056	NN	O	O
bp	NN	O	O
.	NN	O	O

Gel-shift	NN	O	O
assays	NN	O	O
revealed	NN	O	O
that	NN	O	O
transcriptional	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
induced	NN	O	O
only	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-13	NN	O	B-protein
treatment	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
bind	NN	O	O
to	NN	O	O
the	NN	O	O
15-LO	NN	O	B-DNA
promoter	NN	O	I-DNA
DNA	NN	O	I-DNA
.	NN	O	O

Two	NN	O	O
regions	NN	O	O
,	NN	O	O
DP1	NN	O	B-DNA
(	NN	O	O
-140	NN	O	B-DNA
to	NN	O	I-DNA
-92	NN	O	I-DNA
bp	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
DP2	NN	O	B-DNA
(	NN	O	O
-353	NN	O	B-DNA
to	NN	O	I-DNA
-304	NN	O	I-DNA
bp	NN	O	I-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	O
were	NN	O	O
essential	NN	O	O
for	NN	O	O
transcription	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Hela	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
contained	NN	O	O
a	NN	O	O
specific	NN	O	B-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
s	NN	O	I-protein
)	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
15-LO	NN	O	B-DNA
promoter	NN	O	I-DNA
DNA	NN	O	I-DNA
which	NN	O	O
are	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
derived	NN	O	O
from	NN	O	O
IL-13-treated	NN	O	B-cell_line
human	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
monocyte	NN	O	I-cell_line
nuclear	NN	O	O
extracts	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
fluorescent	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
results	NN	O	O
refined	NN	O	O
the	NN	O	O
previous	NN	O	O
localization	NN	O	O
of	NN	O	O
15-LO	NN	O	B-protein
to	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
17p13.3	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Ex	NN	O	O
vivo	NN	O	O
activation	NN	O	O
of	NN	O	O
tumor-draining	NN	O	B-cell_type
lymph	NN	O	I-cell_type
node	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
reverses	NN	O	O
defects	NN	O	O
in	NN	O	O
signal	NN	O	B-protein
transduction	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

The	NN	O	O
adoptive	NN	O	O
transfer	NN	O	O
of	NN	O	O
tumor-draining	NN	O	O
lymph	NN	O	O
node	NN	O	O
(	NN	O	O
LN	NN	O	O
)	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
activated	NN	O	O
ex	NN	O	O
vivo	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
and	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
mediates	NN	O	O
the	NN	O	O
regression	NN	O	O
of	NN	O	O
the	NN	O	O
poorly	NN	O	O
immunogenic	NN	O	O
murine	NN	O	O
melanoma	NN	O	O
D5	NN	O	O
.	NN	O	O

The	NN	O	O
efficacy	NN	O	O
of	NN	O	O
the	NN	O	O
activated	NN	O	B-cell_type
LN	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
augmented	NN	O	O
when	NN	O	O
the	NN	O	O
sensitizing	NN	O	O
tumor	NN	O	O
is	NN	O	O
a	NN	O	O
genetically	NN	O	B-cell_line
modified	NN	O	I-cell_line
variant	NN	O	I-cell_line
(	NN	O	O
designated	NN	O	O
D5G6	NN	O	B-cell_line
)	NN	O	O
that	NN	O	O
secretes	NN	O	O
granulocyte/macrophage-colony-stimulating	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
anti-CD3/IL-2-activated	NN	O	B-cell_line
LN	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
adoptive	NN	O	O
transfer	NN	O	O
of	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
tumor-draining	NN	O	B-cell_type
LN	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
no	NN	O	O
therapeutic	NN	O	O
activity	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
acquisition	NN	O	O
of	NN	O	O
antitumor	NN	O	O
function	NN	O	O
during	NN	O	O
ex	NN	O	O
vivo	NN	O	O
activation	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
modifications	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
capacity	NN	O	O
,	NN	O	O
the	NN	O	O
protein	NN	O	O
tyrosine	NN	O	B-protein
kinases	NN	O	I-protein
p56lck	NN	O	B-protein
and	NN	O	O
p59fyn	NN	O	B-protein
and	NN	O	O
proteins	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB	NN	O	B-protein
family	NN	O	I-protein
were	NN	O	O
analyzed	NN	O	O
in	NN	O	O
tumor-draining	NN	O	B-cell_type
LN	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
levels	NN	O	O
of	NN	O	O
p56lck	NN	O	B-protein
and	NN	O	O
p59fyn	NN	O	B-protein
were	NN	O	O
lower	NN	O	O
in	NN	O	O
tumor-draining	NN	O	O
than	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
LN	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
production	NN	O	O
of	NN	O	O
tyrosine-phosphorylated	NN	O	B-protein
substrates	NN	O	I-protein
was	NN	O	O
markedly	NN	O	O
depressed	NN	O	O
following	NN	O	O
anti-CD3	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

After	NN	O	O
5-day	NN	O	O
anti-CD3	NN	O	B-protein
/IL-2	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
levels	NN	O	O
of	NN	O	O
p56lck	NN	O	B-protein
and	NN	O	O
p59fyn	NN	O	B-protein
and	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
activity	NN	O	O
increased	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
p56lck	NN	O	B-protein
,	NN	O	O
p59fyn	NN	O	B-protein
,	NN	O	O
and	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
activity	NN	O	O
were	NN	O	O
higher	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
derived	NN	O	O
from	NN	O	O
LN	NN	O	O
that	NN	O	O
drained	NN	O	O
D5G6	NN	O	O
than	NN	O	O
they	NN	O	O
were	NN	O	O
in	NN	O	O
those	NN	O	O
from	NN	O	O
D5	NN	O	O
tumors	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
levels	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	O
Rel	NN	O	B-protein
A	NN	O	I-protein
were	NN	O	O
normal	NN	O	O
in	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
tumor-draining	NN	O	O
LN	NN	O	O
,	NN	O	O
as	NN	O	O
was	NN	O	O
nuclear	NN	O	O
kappaB	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
induced	NN	O	O
by	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
or	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
LN	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
D5	NN	O	B-cell_type
tumor	NN	O	I-cell_type
cells	NN	O	I-cell_type
induced	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
recovery	NN	O	O
of	NN	O	O
proteins	NN	O	O
mediating	NN	O	O
signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor/CD3	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
LN	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
activated	NN	O	O
ex	NN	O	O
vivo	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
acquisition	NN	O	O
of	NN	O	O
antitumor	NN	O	O
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
human	NN	O	O
anti-porcine	NN	O	O
T-cell	NN	O	O
immune	NN	O	O
responses	NN	O	O
by	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
transactivator	NN	O	I-DNA
constructs	NN	O	I-DNA
lacking	NN	O	O
the	NN	O	O
amino	NN	O	O
terminal	NN	O	O
domain	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
:	NN	O	O
The	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
transactivator	NN	O	I-protein
(	NN	O	O
CIITA	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
bi-	NN	O	B-protein
or	NN	O	I-protein
multifunctional	NN	O	I-protein
domain	NN	O	I-protein
protein	NN	O	I-protein
that	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
and	NN	O	O
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
a	NN	O	O
mutated	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CIITA	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
coding	NN	O	O
for	NN	O	O
a	NN	O	O
protein	NN	O	O
lacking	NN	O	O
the	NN	O	O
amino	NN	O	B-protein
terminal	NN	O	I-protein
151	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
,	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
potent	NN	O	O
dominant-negative	NN	O	O
suppressor	NN	O	O
of	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Porcine	NN	O	B-protein
MHC	NN	O	I-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
are	NN	O	O
potent	NN	O	O
stimulators	NN	O	O
of	NN	O	O
direct	NN	O	O
T-cell	NN	O	O
recognition	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
are	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
likely	NN	O	O
to	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
rejection	NN	O	O
responses	NN	O	O
to	NN	O	O
transgenic	NN	O	O
pig	NN	O	O
donors	NN	O	O
in	NN	O	O
clinical	NN	O	O
xenotransplantation	NN	O	O
.	NN	O	O

We	NN	O	O
were	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
interested	NN	O	O
in	NN	O	O
examining	NN	O	O
mutated	NN	O	O
CIITA	NN	O	B-DNA
constructs	NN	O	I-DNA
for	NN	O	O
their	NN	O	O
effect	NN	O	O
on	NN	O	O
porcine	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Stable	NN	O	O
transfectants	NN	O	O
of	NN	O	O
the	NN	O	O
porcine	NN	O	B-cell_line
vascular	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
PIEC	NN	O	B-cell_line
with	NN	O	O
mutated	NN	O	O
CIITA	NN	O	B-DNA
constructs	NN	O	I-DNA
were	NN	O	O
tested	NN	O	O
for	NN	O	O
SLA-DR	NN	O	B-protein
and	NN	O	O
SLA-DQ	NN	O	B-protein
induction	NN	O	O
by	NN	O	O
recombinant	NN	O	B-protein
porcine	NN	O	I-protein
interferon-gamma	NN	O	I-protein
.	NN	O	O

Transient	NN	O	O
transfectants	NN	O	O
of	NN	O	O
the	NN	O	O
porcine	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
L23	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
mutated	NN	O	O
CIITA	NN	O	B-DNA
constructs	NN	O	I-DNA
were	NN	O	O
tested	NN	O	O
for	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
constitutive	NN	O	O
SLA-DR	NN	O	O
and	NN	O	O
SLA-DQ	NN	O	O
expression	NN	O	O
.	NN	O	O

T-cell	NN	O	O
proliferation	NN	O	O
studies	NN	O	O
were	NN	O	O
performed	NN	O	O
using	NN	O	O
highly	NN	O	O
purified	NN	O	O
human	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
In	NN	O	O
preliminary	NN	O	O
studies	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
transfection	NN	O	O
of	NN	O	O
the	NN	O	O
PIEC	NN	O	B-cell_line
line	NN	O	I-cell_line
with	NN	O	O
full-length	NN	O	O
human	NN	O	O
CIITA	NN	O	B-DNA
constructs	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
strong	NN	O	O
expression	NN	O	O
of	NN	O	O
SLA-DR	NN	O	B-protein
and	NN	O	O
SLA-DQ	NN	O	B-protein
antigens	NN	O	O
,	NN	O	O
thus	NN	O	O
establishing	NN	O	O
the	NN	O	O
cross-species	NN	O	O
effectiveness	NN	O	O
of	NN	O	O
human	NN	O	B-protein
CIITA	NN	O	I-protein
in	NN	O	O
the	NN	O	O
pig	NN	O	O
.	NN	O	O

The	NN	O	O
mutated	NN	O	B-DNA
human	NN	O	I-DNA
CIITA	NN	O	I-DNA
constructs	NN	O	I-DNA
were	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
tested	NN	O	O
in	NN	O	O
the	NN	O	O
pig	NN	O	O
.	NN	O	O

PIEC	NN	O	B-cell_line
clones	NN	O	I-cell_line
stably	NN	O	O
transfected	NN	O	O
with	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
constructs	NN	O	O
showed	NN	O	O
up	NN	O	O
to	NN	O	O
99	NN	O	O
%	NN	O	O
suppression	NN	O	O
of	NN	O	O
SLA-DR	NN	O	B-protein
and	NN	O	O
SLA-DQ	NN	O	B-protein
antigen	NN	O	O
induction	NN	O	O
and	NN	O	O
marked	NN	O	O
suppression	NN	O	O
of	NN	O	O
SLA-DRA	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
the	NN	O	O
porcine	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
L23	NN	O	I-cell_line
showed	NN	O	O
up	NN	O	O
to	NN	O	O
90	NN	O	O
%	NN	O	O
suppression	NN	O	O
of	NN	O	O
constitutive	NN	O	O
SLA-DR	NN	O	O
and	NN	O	O
SLA-DQ	NN	O	O
antigen	NN	O	O
expression	NN	O	O
in	NN	O	O
5-8	NN	O	O
days	NN	O	O
.	NN	O	O

In	NN	O	O
functional	NN	O	O
studies	NN	O	O
,	NN	O	O
interferon-gamma-stimulated	NN	O	B-cell_line
PIEC	NN	O	I-cell_line
clones	NN	O	I-cell_line
transfected	NN	O	O
with	NN	O	O
this	NN	O	O
mutated	NN	O	B-protein
CIITA	NN	O	I-protein
construct	NN	O	O
failed	NN	O	O
to	NN	O	O
stimulate	NN	O	O
purified	NN	O	O
human	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Mutated	NN	O	B-DNA
human	NN	O	I-DNA
CIITA	NN	O	I-DNA
constructs	NN	O	I-DNA
are	NN	O	O
potent	NN	O	O
suppressors	NN	O	O
of	NN	O	O
porcine	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Macrophages	NN	O	B-cell_type
in	NN	O	O
human	NN	O	O
atheroma	NN	O	O
contain	NN	O	O
PPARgamma	NN	O	B-protein
:	NN	O	O
differentiation-dependent	NN	O	B-protein
peroxisomal	NN	O	I-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
gamma	NN	O	I-protein
(	NN	O	O
PPARgamma	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
and	NN	O	O
reduction	NN	O	O
of	NN	O	O
MMP-9	NN	O	B-protein
activity	NN	O	O
through	NN	O	O
PPARgamma	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
mononuclear	NN	O	O
phagocytes	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
atherosclerosis	NN	O	O
and	NN	O	O
its	NN	O	O
sequela	NN	O	O
plaque	NN	O	O
rupture	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
their	NN	O	O
secretion	NN	O	O
of	NN	O	O
matrix	NN	O	B-protein
metalloproteinases	NN	O	I-protein
(	NN	O	O
MMPs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
including	NN	O	O
MMP-9	NN	O	B-protein
.	NN	O	O

Peroxisomal	NN	O	B-protein
proliferator-activated	NN	O	I-protein
receptor	NN	O	I-protein
gamma	NN	O	I-protein
(	NN	O	O
PPARgamma	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
receptor	NN	O	O
superfamily	NN	O	O
,	NN	O	O
regulates	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
various	NN	O	O
activators	NN	O	O
,	NN	O	O
including	NN	O	O
15-deoxy-delta12	NN	O	O
,	NN	O	O
14-prostaglandin	NN	O	O
J2	NN	O	O
and	NN	O	O
the	NN	O	O
antidiabetic	NN	O	O
agent	NN	O	O
troglitazone	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
PPARgamma	NN	O	B-protein
in	NN	O	O
human	NN	O	O
atherosclerosis	NN	O	O
is	NN	O	O
unexplored	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
in	NN	O	O
human	NN	O	O
atherosclerotic	NN	O	O
lesions	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
12	NN	O	O
)	NN	O	O
express	NN	O	O
immunostainable	NN	O	B-protein
PPARgamma	NN	O	I-protein
.	NN	O	O

Normal	NN	O	O
artery	NN	O	O
specimens	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
6	NN	O	O
)	NN	O	O
reveal	NN	O	O
minimal	NN	O	B-protein
immunoreactive	NN	O	I-protein
PPARgamma	NN	O	I-protein
.	NN	O	O

Human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
cultured	NN	O	O
for	NN	O	O
6	NN	O	O
days	NN	O	O
in	NN	O	O
5	NN	O	O
%	NN	O	O
human	NN	O	O
serum	NN	O	O
expressed	NN	O	O
PPARgamma	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
by	NN	O	O
reverse	NN	O	O
transcription-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
and	NN	O	O
Western	NN	O	O
blotting	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
PPARgamma	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
increased	NN	O	O
during	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13	NN	O	O
acetate-induced	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
PPARgamma	NN	O	B-protein
with	NN	O	O
troglitazone	NN	O	O
or	NN	O	O
15-deoxy-delta12	NN	O	O
,	NN	O	O
14-prostaglandin	NN	O	O
J2	NN	O	O
in	NN	O	O
human	NN	O	O
monocyte-derived	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
inhibited	NN	O	O
MMP-9	NN	O	B-protein
gelatinolytic	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
concentration-dependent	NN	O	O
fashion	NN	O	O
as	NN	O	O
revealed	NN	O	O
by	NN	O	O
zymography	NN	O	O
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
correlates	NN	O	O
with	NN	O	O
decreased	NN	O	O
MMP-9	NN	O	B-protein
secretion	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
Western	NN	O	O
blotting	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
PPARgamma	NN	O	B-protein
is	NN	O	O
present	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
in	NN	O	O
human	NN	O	O
atherosclerotic	NN	O	O
lesions	NN	O	O
and	NN	O	O
may	NN	O	O
regulate	NN	O	O
expression	NN	O	O
and	NN	O	O
activity	NN	O	O
of	NN	O	O
MMP-9	NN	O	B-protein
,	NN	O	O
an	NN	O	O
enzyme	NN	O	O
implicated	NN	O	O
in	NN	O	O
plaque	NN	O	O
rupture	NN	O	O
.	NN	O	O

PPARgamma	NN	O	B-protein
is	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
regulator	NN	O	O
of	NN	O	O
monocyte/macrophage	NN	O	O
function	NN	O	O
with	NN	O	O
relevance	NN	O	O
for	NN	O	O
human	NN	O	O
atherosclerotic	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Erythropoietin	NN	O	B-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak2	NN	O	B-protein
,	NN	O	O
STAT5A	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT5B	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
cultured	NN	O	I-cell_type
human	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
precursors	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
examined	NN	O	O
signaling	NN	O	O
by	NN	O	O
erythropoietin	NN	O	B-protein
in	NN	O	O
highly	NN	O	B-cell_type
purified	NN	O	I-cell_type
human	NN	O	I-cell_type
colony	NN	O	I-cell_type
forming	NN	O	I-cell_type
unit-erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
generated	NN	O	O
in	NN	O	O
vitro	NN	O	O
from	NN	O	O
CD34	NN	O	B-cell_type
(	NN	O	I-cell_type
+	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
erythropoietin	NN	O	B-protein
induces	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak2	NN	O	B-protein
,	NN	O	O
STAT5A	NN	O	B-protein
,	NN	O	O
and	NN	O	O
STAT5B	NN	O	B-protein
.	NN	O	O

Tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak2	NN	O	B-protein
reaches	NN	O	O
a	NN	O	O
peak	NN	O	O
around	NN	O	O
10	NN	O	O
minutes	NN	O	O
after	NN	O	O
stimulation	NN	O	O
and	NN	O	O
is	NN	O	O
maximum	NN	O	O
at	NN	O	O
5	NN	O	O
U/mL	NN	O	O
of	NN	O	O
erythropoietin	NN	O	B-protein
.	NN	O	O

Tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT5	NN	O	B-protein
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
the	NN	O	O
translocation	NN	O	O
of	NN	O	O
activated	NN	O	O
STAT5	NN	O	B-protein
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
using	NN	O	O
32Pi-labeled	NN	O	O
STAT5	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
beta-casein	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Tyrosine	NN	O	O
phosphorylation	NN	O	O
STAT1	NN	O	B-protein
or	NN	O	O
STAT3	NN	O	B-protein
was	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
precursors	NN	O	I-cell_type
after	NN	O	O
stimulation	NN	O	O
with	NN	O	O
erythropoietin	NN	O	B-protein
.	NN	O	O

Crkl	NN	O	B-protein
,	NN	O	O
an	NN	O	O
SH2/SH3	NN	O	B-protein
adapter	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
becomes	NN	O	O
coimmunoprecipitated	NN	O	O
specifically	NN	O	O
with	NN	O	O
STAT5	NN	O	B-protein
from	NN	O	O
erythropoietin-stimulated	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
although	NN	O	O
it	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
become	NN	O	O
associated	NN	O	O
with	NN	O	O
c-Cbl	NN	O	B-protein
in	NN	O	O
the	NN	O	O
studies	NN	O	O
using	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
human	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
precursors	NN	O	I-cell_type
can	NN	O	O
be	NN	O	O
expanded	NN	O	O
in	NN	O	O
vitro	NN	O	O
in	NN	O	O
sufficient	NN	O	O
numbers	NN	O	O
and	NN	O	O
purity	NN	O	O
to	NN	O	O
allow	NN	O	O
its	NN	O	O
usage	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
studies	NN	O	O
.	NN	O	O

This	NN	O	O
report	NN	O	O
sets	NN	O	O
a	NN	O	O
basis	NN	O	O
for	NN	O	O
further	NN	O	O
studies	NN	O	O
on	NN	O	O
signaling	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
cultured	NN	O	I-cell_type
human	NN	O	I-cell_type
erythroid	NN	O	I-cell_type
precursors	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
contribute	NN	O	O
to	NN	O	O
our	NN	O	O
better	NN	O	O
understanding	NN	O	O
in	NN	O	O
the	NN	O	O
differentiation	NN	O	O
processes	NN	O	O
of	NN	O	O
erythrocytes	NN	O	B-cell_type
and	NN	O	O
their	NN	O	O
precursors	NN	O	O
.	NN	O	O

-DOCSTART-	O

Oxidative	NN	O	O
stress	NN	O	O
suppresses	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
activities	NN	O	O
in	NN	O	O
stimulated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Effects	NN	O	O
of	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
on	NN	O	O
stimulation-dependent	NN	O	O
signal	NN	O	O
transduction	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

Purified	NN	O	B-cell_type
quiescent	NN	O	I-cell_type
human	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
subjected	NN	O	O
to	NN	O	O
:	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
acute	NN	O	O
exposure	NN	O	O
to	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
chronic	NN	O	O
exposure	NN	O	O
to	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
acute	NN	O	O
exposure	NN	O	O
to	NN	O	O
ionizing	NN	O	O
radiation	NN	O	O
.	NN	O	O

The	NN	O	O
cells	NN	O	O
were	NN	O	O
then	NN	O	O
stimulated	NN	O	O
for	NN	O	O
6	NN	O	O
h	NN	O	O
.	NN	O	O

DNA-binding	NN	O	O
activities	NN	O	O
(	NN	O	O
determined	NN	O	O
by	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
)	NN	O	O
of	NN	O	O
three	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
:	NN	O	O
NFkappaB	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
were	NN	O	O
abolished	NN	O	O
in	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
by	NN	O	O
all	NN	O	O
three	NN	O	O
modes	NN	O	O
of	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
.	NN	O	O

The	NN	O	O
lymphocytes	NN	O	B-cell_type
exhibited	NN	O	O
lipid	NN	O	O
peroxidation	NN	O	O
only	NN	O	O
upon	NN	O	O
exposure	NN	O	O
to	NN	O	O
the	NN	O	O
lowest	NN	O	O
level	NN	O	O
of	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
used	NN	O	O
(	NN	O	O
20	NN	O	O
microM	NN	O	O
)	NN	O	O
.	NN	O	O

All	NN	O	O
three	NN	O	O
modes	NN	O	O
of	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
induced	NN	O	O
catalase	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
only	NN	O	O
exception	NN	O	O
was	NN	O	O
hydrogen	NN	O	O
peroxide	NN	O	O
at	NN	O	O
20	NN	O	O
microM	NN	O	O
,	NN	O	O
which	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
catalase	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
:	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
suppression	NN	O	O
of	NN	O	O
specific	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
functions	NN	O	O
can	NN	O	O
potentially	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
marker	NN	O	O
of	NN	O	O
exposure	NN	O	O
to	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
and	NN	O	O
its	NN	O	O
effects	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
lipid	NN	O	O
peroxidation	NN	O	O
is	NN	O	O
only	NN	O	O
detectable	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
upon	NN	O	O
exposure	NN	O	O
to	NN	O	O
weak	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
which	NN	O	O
does	NN	O	O
not	NN	O	O
induce	NN	O	O
catalase	NN	O	B-protein
activity	NN	O	O
;	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
therefore	NN	O	O
,	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
DNA-binding	NN	O	O
activities	NN	O	O
are	NN	O	O
more	NN	O	O
sensitive	NN	O	O
to	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
than	NN	O	O
lipid	NN	O	O
peroxidation	NN	O	O
.	NN	O	O

-DOCSTART-	O

IL-4	NN	O	B-protein
-dependent	NN	O	O
regulation	NN	O	O
of	NN	O	O
TGF-alpha	NN	O	O
and	NN	O	O
TGF-beta1	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
.	NN	O	O

TGFs	NN	O	B-protein
play	NN	O	O
important	NN	O	O
roles	NN	O	O
in	NN	O	O
wound	NN	O	O
healing	NN	O	O
and	NN	O	O
carcinogenesis	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
eosinophils	NN	O	B-cell_type
infiltrating	NN	O	O
into	NN	O	O
different	NN	O	O
pathologic	NN	O	O
processes	NN	O	O
elaborate	NN	O	O
TGF-alpha	NN	O	B-protein
and	NN	O	O
TGF-beta1	NN	O	B-protein
.	NN	O	O

Eosinophils	NN	O	B-cell_type
infiltrating	NN	O	O
hamster	NN	O	O
cutaneous	NN	O	O
wounds	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
express	NN	O	O
TGFs	NN	O	B-protein
sequentially	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
biologic	NN	O	O
mediators	NN	O	O
that	NN	O	O
may	NN	O	O
regulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
TGF-alpha	NN	O	B-protein
and	NN	O	I-protein
-beta1	NN	O	I-protein
by	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

Eosinophils	NN	O	B-cell_type
were	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
healthy	NN	O	O
donors	NN	O	O
and	NN	O	O
cultured	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
or	NN	O	O
presence	NN	O	O
of	NN	O	O
IL-3	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-5	NN	O	B-protein
.	NN	O	O

Cells	NN	O	O
were	NN	O	O
analyzed	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
and	NN	O	O
immunohistochemistry	NN	O	O
.	NN	O	O

Supernatants	NN	O	O
from	NN	O	O
these	NN	O	O
cultures	NN	O	O
were	NN	O	O
assayed	NN	O	O
for	NN	O	O
secreted	NN	O	O
TGF-alpha	NN	O	B-protein
and	NN	O	O
TGF-beta1	NN	O	B-protein
using	NN	O	O
TGF	NN	O	B-protein
-specific	NN	O	O
ELISAs	NN	O	O
.	NN	O	O

IL-3	NN	O	B-protein
,	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-5	NN	O	B-protein
independently	NN	O	O
up-regulated	NN	O	O
TGF-beta1	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
product	NN	O	O
expression	NN	O	O
by	NN	O	O
eosinophils	NN	O	B-cell_type
in	NN	O	O
all	NN	O	O
donors	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
TGF-alpha	NN	O	B-protein
production	NN	O	O
by	NN	O	O
eosinophils	NN	O	B-cell_type
was	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
IL-3	NN	O	B-protein
and	NN	O	O
IL-5	NN	O	B-protein
but	NN	O	O
was	NN	O	O
down-regulated	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
to	NN	O	O
regulate	NN	O	O
eosinophil	NN	O	O
responses	NN	O	O
,	NN	O	O
IL-4	NN	O	B-protein
signaling	NN	O	I-protein
molecules	NN	O	I-protein
are	NN	O	O
present	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
observation	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
can	NN	O	O
differentially	NN	O	O
regulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
TGF-alpha	NN	O	B-protein
and	NN	O	O
TGF-beta1	NN	O	B-protein
suggests	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
may	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
physiologic	NN	O	O
molecular	NN	O	O
switch	NN	O	O
of	NN	O	O
TGF	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
the	NN	O	O
infiltrating	NN	O	O
eosinophils	NN	O	B-cell_type
in	NN	O	O
wound	NN	O	O
healing	NN	O	O
and	NN	O	O
carcinogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

MLL	NN	O	O
and	NN	O	O
CALM	NN	O	B-DNA
are	NN	O	O
fused	NN	O	O
to	NN	O	O
AF10	NN	O	B-DNA
in	NN	O	O
morphologically	NN	O	O
distinct	NN	O	O
subsets	NN	O	O
of	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
with	NN	O	O
translocation	NN	O	B-DNA
t	NN	O	I-DNA
(	NN	O	I-DNA
10	NN	O	I-DNA
;	NN	O	I-DNA
11	NN	O	I-DNA
)	NN	O	I-DNA
:	NN	O	O
both	NN	O	O
rearrangements	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
poor	NN	O	O
prognosis	NN	O	O
.	NN	O	O

The	NN	O	O
translocation	NN	O	B-DNA
t	NN	O	I-DNA
(	NN	O	I-DNA
10	NN	O	I-DNA
;	NN	O	I-DNA
11	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
p13	NN	O	I-DNA
;	NN	O	I-DNA
q14	NN	O	I-DNA
)	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ALL	NN	O	O
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
(	NN	O	O
AML	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
recent	NN	O	O
study	NN	O	O
showed	NN	O	O
a	NN	O	O
MLL/AF10	NN	O	B-DNA
fusion	NN	O	I-DNA
in	NN	O	O
all	NN	O	O
cases	NN	O	O
of	NN	O	O
AML	NN	O	O
with	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
10	NN	O	I-DNA
;	NN	O	I-DNA
11	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
various	NN	O	O
breakpoints	NN	O	O
on	NN	O	O
chromosome	NN	O	B-DNA
11	NN	O	I-DNA
ranging	NN	O	O
from	NN	O	O
q13	NN	O	B-DNA
to	NN	O	O
q23	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
recently	NN	O	O
cloned	NN	O	O
CALM	NN	O	B-DNA
(	NN	O	O
Clathrin	NN	O	B-DNA
Assembly	NN	O	I-DNA
Lymphoid	NN	O	I-DNA
Myeloid	NN	O	I-DNA
leukemia	NN	O	I-DNA
gene	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
the	NN	O	O
fusion	NN	O	O
partner	NN	O	O
of	NN	O	O
AF10	NN	O	B-DNA
at	NN	O	O
11q14	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U937	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
further	NN	O	O
define	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
in	NN	O	O
acute	NN	O	O
leukemias	NN	O	O
,	NN	O	O
10	NN	O	O
cases	NN	O	O
(	NN	O	O
9	NN	O	O
AML	NN	O	O
and	NN	O	O
1	NN	O	O
ALL	NN	O	O
)	NN	O	O
with	NN	O	O
cytogenetically	NN	O	O
proven	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
10	NN	O	I-DNA
;	NN	O	I-DNA
11	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
p12-14	NN	O	I-DNA
;	NN	O	I-DNA
q13-21	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
well-characterized	NN	O	O
morphology	NN	O	O
,	NN	O	O
immunophenotype	NN	O	O
,	NN	O	O
and	NN	O	O
clinical	NN	O	O
course	NN	O	O
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

Interphase	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
(	NN	O	O
FISH	NN	O	O
)	NN	O	O
was	NN	O	O
performed	NN	O	O
with	NN	O	O
2	NN	O	O
YACs	NN	O	B-DNA
flanking	NN	O	O
the	NN	O	O
CALM	NN	O	B-DNA
region	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
YAC	NN	O	B-DNA
contig	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
MLL	NN	O	B-DNA
region	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
YAC	NN	O	B-DNA
spanning	NN	O	O
the	NN	O	O
AF10	NN	O	B-DNA
breakpoint	NN	O	O
.	NN	O	O

Rearrangement	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
all	NN	O	O
cases	NN	O	O
with	NN	O	O
balanced	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
10	NN	O	I-DNA
;	NN	O	I-DNA
11	NN	O	I-DNA
)	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
4	NN	O	O
cases	NN	O	O
,	NN	O	O
including	NN	O	O
3	NN	O	O
AML	NN	O	O
with	NN	O	O
immature	NN	O	O
morphology	NN	O	O
(	NN	O	O
1	NN	O	O
AML-M0	NN	O	O
and	NN	O	O
2	NN	O	O
AML-M1	NN	O	O
)	NN	O	O
and	NN	O	O
1	NN	O	O
ALL	NN	O	O
,	NN	O	O
the	NN	O	O
signals	NN	O	O
of	NN	O	O
the	NN	O	O
CALM	NN	O	B-DNA
YACS	NN	O	I-DNA
were	NN	O	O
separated	NN	O	O
in	NN	O	O
interphase	NN	O	O
cells	NN	O	O
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
translocation	NN	O	O
breakpoint	NN	O	O
within	NN	O	O
the	NN	O	O
CALM	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

MLL	NN	O	O
was	NN	O	O
rearranged	NN	O	O
in	NN	O	O
3	NN	O	O
AML	NN	O	O
with	NN	O	O
myelomonocytic	NN	O	O
differentiation	NN	O	O
(	NN	O	O
2	NN	O	O
AML-M2	NN	O	O
and	NN	O	O
1	NN	O	O
AML-M5	NN	O	O
)	NN	O	O
,	NN	O	O
including	NN	O	O
1	NN	O	O
secondary	NN	O	O
AML	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
3	NN	O	O
cases	NN	O	O
,	NN	O	O
a	NN	O	O
characteristic	NN	O	O
immunophenotype	NN	O	O
was	NN	O	O
identified	NN	O	O
(	NN	O	O
CD4+	NN	O	B-protein
,	NN	O	O
CD13	NN	O	B-protein
-	NN	O	O
,	NN	O	O
CD33+	NN	O	O
,	NN	O	O
CD65s+	NN	O	O
)	NN	O	O
.	NN	O	O

AF-10	NN	O	O
was	NN	O	O
involved	NN	O	O
in	NN	O	O
5	NN	O	O
of	NN	O	O
6	NN	O	O
evaluable	NN	O	O
cases	NN	O	O
,	NN	O	O
including	NN	O	O
1	NN	O	O
case	NN	O	O
without	NN	O	O
detectable	NN	O	O
CALM	NN	O	B-DNA
or	NN	O	O
MLL	NN	O	O
rearrangement	NN	O	O
.	NN	O	O

In	NN	O	O
2	NN	O	O
complex	NN	O	O
translocations	NN	O	O
,	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
genes	NN	O	O
was	NN	O	O
rearranged	NN	O	O
.	NN	O	O

All	NN	O	O
cases	NN	O	O
had	NN	O	O
a	NN	O	O
remarkably	NN	O	O
poor	NN	O	O
prognosis	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
mean	NN	O	O
survival	NN	O	O
of	NN	O	O
9.6	NN	O	O
+/-	NN	O	O
6.6	NN	O	O
months	NN	O	O
.	NN	O	O

For	NN	O	O
the	NN	O	O
7	NN	O	O
AML	NN	O	O
cases	NN	O	O
that	NN	O	O
were	NN	O	O
uniformly	NN	O	O
treated	NN	O	O
according	NN	O	O
to	NN	O	O
the	NN	O	O
AMLCG86/92	NN	O	O
protocols	NN	O	O
,	NN	O	O
disease-free	NN	O	O
and	NN	O	O
overall	NN	O	O
survival	NN	O	O
was	NN	O	O
significantly	NN	O	O
worse	NN	O	O
than	NN	O	O
for	NN	O	O
the	NN	O	O
overall	NN	O	O
study	NN	O	O
group	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.03	NN	O	O
and	NN	O	O
P	NN	O	O
=	NN	O	O
.01	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
10	NN	O	I-DNA
;	NN	O	I-DNA
11	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
p13	NN	O	I-DNA
;	NN	O	I-DNA
q14	NN	O	I-DNA
)	NN	O	I-DNA
indicates	NN	O	O
CALM	NN	O	B-DNA
and	NN	O	I-DNA
MLL	NN	O	I-DNA
rearrangements	NN	O	I-DNA
in	NN	O	O
morphologically	NN	O	O
distinct	NN	O	O
subsets	NN	O	O
of	NN	O	O
acute	NN	O	O
leukemia	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
poor	NN	O	O
prognosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
E-proteins	NN	O	B-protein
in	NN	O	O
B-	NN	O	O
and	NN	O	O
T-lymphocyte	NN	O	O
development	NN	O	O
.	NN	O	O

Department	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
hematopoietic	NN	O	B-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
controlled	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
one	NN	O	O
class	NN	O	O
of	NN	O	O
helix-loop-helix	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
termed	NN	O	O
E-proteins	NN	O	B-protein
,	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
B-cell	NN	O	O
development	NN	O	O
.	NN	O	O

Recent	NN	O	O
analysis	NN	O	O
of	NN	O	O
gene-targeted	NN	O	O
mice	NN	O	O
has	NN	O	O
allowed	NN	O	O
a	NN	O	O
direct	NN	O	O
assessment	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
roles	NN	O	O
of	NN	O	O
several	NN	O	B-protein
E-protein	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
in	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
review	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
defects	NN	O	O
in	NN	O	O
B-	NN	O	O
and	NN	O	O
T-	NN	O	O
lymphocyte	NN	O	O
development	NN	O	O
in	NN	O	O
mice	NN	O	O
carrying	NN	O	O
targeted	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
E-protein	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
discuss	NN	O	O
our	NN	O	O
current	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
development	NN	O	O

-DOCSTART-	O

Epstein-Barr	NN	O	B-protein
virus-transforming	NN	O	I-protein
protein	NN	O	I-protein
latent	NN	O	B-protein
infection	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
activates	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	I-protein
through	NN	O	O
a	NN	O	O
pathway	NN	O	O
that	NN	O	O
includes	NN	O	O
the	NN	O	O
NF-kappaB-inducing	NN	O	B-protein
kinase	NN	O	I-protein
and	NN	O	O
the	NN	O	O
IkappaB	NN	O	B-protein
kinases	NN	O	I-protein
IKKalpha	NN	O	B-protein
and	NN	O	O
IKKbeta	NN	O	B-protein
.	NN	O	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
oncoprotein	NN	O	I-protein
latent	NN	O	B-protein
infection	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
LMP1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
constitutively	NN	O	O
aggregated	NN	O	O
pseudo-tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TNFR	NN	O	B-protein
)	NN	O	O
that	NN	O	O
activates	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	I-protein
through	NN	O	O
two	NN	O	O
sites	NN	O	O
in	NN	O	O
its	NN	O	O
C-terminal	NN	O	B-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

One	NN	O	O
site	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
activated	NN	O	O
TNFRII	NN	O	B-protein
in	NN	O	O
associating	NN	O	O
with	NN	O	O
TNFR-associated	NN	O	B-protein
factors	NN	O	I-protein
TRAF1	NN	O	B-protein
and	NN	O	O
TRAF2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
second	NN	O	O
site	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
TNFRI	NN	O	B-protein
in	NN	O	O
associating	NN	O	O
with	NN	O	O
the	NN	O	O
TNFRI	NN	O	B-protein
death	NN	O	I-protein
domain	NN	O	I-protein
interacting	NN	O	I-protein
protein	NN	O	I-protein
TRADD	NN	O	B-protein
.	NN	O	O

TNFRI	NN	O	B-protein
has	NN	O	O
been	NN	O	O
recently	NN	O	O
shown	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappaB	NN	O	B-protein
through	NN	O	O
association	NN	O	O
with	NN	O	O
TRADD	NN	O	B-protein
,	NN	O	O
RIP	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TRAF2	NN	O	B-protein
;	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappaB-inducing	NN	O	B-protein
kinase	NN	O	I-protein
(	NN	O	O
NIK	NN	O	B-protein
)	NN	O	O
;	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
kinases	NN	O	I-protein
(	NN	O	O
IKKalpha	NN	O	B-protein
and	NN	O	O
IKKbeta	NN	O	B-protein
)	NN	O	O
;	NN	O	O
and	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

IkappaB	NN	O	B-protein
alpha	NN	O	I-protein
phosphorylation	NN	O	O
on	NN	O	O
Ser-32	NN	O	O
and	NN	O	O
Ser-36	NN	O	O
is	NN	O	O
followed	NN	O	O
by	NN	O	O
its	NN	O	O
degradation	NN	O	O
and	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
LMP1	NN	O	B-protein
or	NN	O	O
by	NN	O	O
each	NN	O	O
of	NN	O	O
its	NN	O	O
effector	NN	O	B-protein
sites	NN	O	I-protein
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
pathway	NN	O	O
that	NN	O	O
includes	NN	O	O
NIK	NN	O	B-protein
,	NN	O	O
IKKalpha	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IKKbeta	NN	O	B-protein
.	NN	O	O

Dominant	NN	O	B-protein
negative	NN	O	I-protein
mutants	NN	O	I-protein
of	NN	O	O
NIK	NN	O	B-protein
,	NN	O	O
IKKalpha	NN	O	B-protein
,	NN	O	O
or	NN	O	O
IKKbeta	NN	O	B-protein
substantially	NN	O	O
inhibited	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
LMP1	NN	O	B-protein
or	NN	O	O
by	NN	O	O
each	NN	O	O
of	NN	O	O
its	NN	O	O
effector	NN	O	B-protein
sites	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Targeted	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
MyD88	NN	O	B-DNA
gene	NN	O	I-DNA
results	NN	O	O
in	NN	O	O
loss	NN	O	O
of	NN	O	O
IL-1-	NN	O	O
and	NN	O	O
IL-18-mediated	NN	O	O
function	NN	O	O
.	NN	O	O

MyD88	NN	O	B-protein
,	NN	O	O
originally	NN	O	O
isolated	NN	O	O
as	NN	O	O
a	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
primary	NN	O	B-DNA
response	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
shown	NN	O	O
to	NN	O	O
act	NN	O	O
as	NN	O	O
an	NN	O	O
adaptor	NN	O	O
in	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
by	NN	O	O
interacting	NN	O	O
with	NN	O	O
both	NN	O	O
the	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
and	NN	O	O
IL-1	NN	O	B-protein
receptor-associated	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
IRAK	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Mice	NN	O	O
generated	NN	O	O
by	NN	O	O
gene	NN	O	O
targeting	NN	O	O
to	NN	O	O
lack	NN	O	O
MyD88	NN	O	B-protein
have	NN	O	O
defects	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
induction	NN	O	O
of	NN	O	O
acute	NN	O	O
phase	NN	O	O
proteins	NN	O	O
and	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-1	NN	O	B-protein
.	NN	O	O

Increases	NN	O	O
in	NN	O	O
interferon-gamma	NN	O	B-protein
production	NN	O	O
and	NN	O	O
natural	NN	O	O
killer	NN	O	O
cell	NN	O	O
activity	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
IL-18	NN	O	B-protein
are	NN	O	O
abrogated	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
Th1	NN	O	O
response	NN	O	O
is	NN	O	O
also	NN	O	O
impaired	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
IL-18	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
N-terminal	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
JNK	NN	O	B-protein
)	NN	O	O
is	NN	O	O
blocked	NN	O	O
in	NN	O	O
MyD88-/-	NN	O	B-cell_line
Th1-developing	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
MyD88	NN	O	B-protein
is	NN	O	O
a	NN	O	O
critical	NN	O	O
component	NN	O	O
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
cascade	NN	O	O
that	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
IL-1	NN	O	B-protein
receptor	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
IL-18	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Ciprofloxacin	NN	O	O
induces	NN	O	O
an	NN	O	O
immunomodulatory	NN	O	O
stress	NN	O	O
response	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
cells	NN	O	O
to	NN	O	O
adverse	NN	O	O
environmental	NN	O	O
conditions	NN	O	O
invokes	NN	O	O
a	NN	O	O
genetically	NN	O	O
programmed	NN	O	O
series	NN	O	O
of	NN	O	O
events	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
fluoroquinolone	NN	O	O
antibiotic	NN	O	O
ciprofloxacin	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
upregulate	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
gene	NN	O	O
induction	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
investigation	NN	O	O
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
ciprofloxacin	NN	O	O
at	NN	O	O
supratherapeutic	NN	O	O
concentrations	NN	O	O
on	NN	O	O
immediate-early	NN	O	O
(	NN	O	O
<	NN	O	O
2	NN	O	O
h	NN	O	O
)	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
was	NN	O	O
studied	NN	O	O
with	NN	O	O
Northern	NN	O	O
blots	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
metallothionein	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	I-DNA
promoter	NN	O	I-DNA
regions	NN	O	I-DNA
and	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP-1	NN	O	B-protein
,	NN	O	O
NF-kappaB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-AT	NN	O	B-protein
were	NN	O	O
analyzed	NN	O	O
by	NN	O	O
chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
(	NN	O	O
CAT	NN	O	B-protein
)	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
concentration	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
,	NN	O	I-RNA
c-jun	NN	O	I-RNA
,	NN	O	I-RNA
c-myc	NN	O	I-RNA
,	NN	O	I-RNA
junB	NN	O	I-RNA
,	NN	O	I-RNA
and	NN	O	I-RNA
fra-1	NN	O	I-RNA
mRNAs	NN	O	I-RNA
was	NN	O	O
increased	NN	O	O
in	NN	O	O
activated	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
incubated	NN	O	O
with	NN	O	O
ciprofloxacin	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
in	NN	O	O
untreated	NN	O	O
controls	NN	O	O
.	NN	O	O

Ciprofloxacin	NN	O	O
increased	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
stimulated	NN	O	O
lymphocytes	NN	O	O
transfected	NN	O	O
with	NN	O	O
plasmids	NN	O	B-cell_type
containing	NN	O	O
either	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
or	NN	O	O
metallothionein	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
among	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
tested	NN	O	O
,	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
increased	NN	O	O
in	NN	O	O
stimulated	NN	O	B-cell_type
purified	NN	O	I-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
incubated	NN	O	O
with	NN	O	O
ciprofloxacin	NN	O	O
compared	NN	O	O
to	NN	O	O
drug-free	NN	O	B-cell_type
controls	NN	O	I-cell_type
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
ciprofloxacin	NN	O	O
increased	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
immediate-early	NN	O	B-RNA
transcripts	NN	O	I-RNA
,	NN	O	O
enhanced	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
metallothionein	NN	O	B-protein
promoter	NN	O	O
induction	NN	O	O
,	NN	O	O
and	NN	O	O
upregulated	NN	O	O
AP-1	NN	O	B-protein
concentrations	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
reflecting	NN	O	O
a	NN	O	O
program	NN	O	O
commonly	NN	O	O
observed	NN	O	O
in	NN	O	O
mammalian	NN	O	O
stress	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
RAR	NN	O	B-protein
)	NN	O	O
-selective	NN	O	O
antagonist	NN	O	O
inhibits	NN	O	O
differentiation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
:	NN	O	O
implications	NN	O	O
of	NN	O	O
RARalpha	NN	O	B-protein
-mediated	NN	O	O
signals	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
induces	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
differentiate	NN	O	O
terminally	NN	O	O
into	NN	O	O
mature	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
subsequently	NN	O	O
die	NN	O	O
by	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

The	NN	O	O
biological	NN	O	O
effects	NN	O	O
of	NN	O	O
RA	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
two	NN	O	O
distinct	NN	O	O
families	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
:	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RARs	NN	O	B-protein
)	NN	O	O
and	NN	O	O
retinoid	NN	O	B-protein
X	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
RXRs	NN	O	B-protein
)	NN	O	O
.	NN	O	O

RARs	NN	O	B-protein
and	NN	O	O
RXRs	NN	O	B-protein
form	NN	O	O
heterodimers	NN	O	B-protein
and	NN	O	O
regulate	NN	O	O
retinoid-mediated	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
developed	NN	O	O
a	NN	O	O
novel	NN	O	O
RAR	NN	O	B-protein
-selective	NN	O	O
antagonist	NN	O	O
(	NN	O	O
ER27191	NN	O	O
)	NN	O	O
which	NN	O	O
prevents	NN	O	O
RAR	NN	O	B-protein
activation	NN	O	O
by	NN	O	O
retinoids	NN	O	O
.	NN	O	O

Using	NN	O	O
this	NN	O	O
RAR-selective	NN	O	O
antagonist	NN	O	O
,	NN	O	O
and	NN	O	O
RXR	NN	O	B-protein
and	NN	O	O
RAR	NN	O	B-protein
agonist	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
RAR	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
is	NN	O	O
important	NN	O	O
for	NN	O	O
differentiation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Simple	NN	O	O
activation	NN	O	O
of	NN	O	O
RXRs	NN	O	B-protein
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
induce	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
the	NN	O	O
RAR-selective	NN	O	O
antagonist	NN	O	O
and	NN	O	O
9-cis	NN	O	O
RA	NN	O	O
resulted	NN	O	O
in	NN	O	O
partial	NN	O	O
differentiation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
and	NN	O	I-cell_line
NB4	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
RAR	NN	O	B-protein
antagonist	NN	O	O
completely	NN	O	O
blocked	NN	O	O
all-trans	NN	O	O
RA-induced	NN	O	O
differentiation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

Additional	NN	O	O
experiments	NN	O	O
showed	NN	O	O
that	NN	O	O
levels	NN	O	O
of	NN	O	O
BCL-2	NN	O	B-protein
protein	NN	O	I-protein
decreased	NN	O	O
during	NN	O	O
differentiation	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
transduced	NN	O	O
with	NN	O	O
a	NN	O	O
bcl-2	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
showed	NN	O	O
the	NN	O	O
same	NN	O	O
differentiation	NN	O	O
response	NN	O	O
to	NN	O	O
retinoids	NN	O	O
as	NN	O	O
did	NN	O	O
parental	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
even	NN	O	O
though	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
inhibited	NN	O	O
in	NN	O	O
these	NN	O	O
bcl-2-transduced	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
differentiation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
are	NN	O	O
regulated	NN	O	O
independently	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Thrombopoietin	NN	O	B-protein
supports	NN	O	O
in	NN	O	O
vitro	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
via	NN	O	O
its	NN	O	O
specific	NN	O	O
receptor	NN	O	O
c-Mpl	NN	O	B-protein
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Thrombopoietin	NN	O	B-protein
(	NN	O	O
TPO	NN	O	B-protein
)	NN	O	O
acts	NN	O	O
on	NN	O	O
megakaryopoiesis	NN	O	O
and	NN	O	O
erythropoiesis	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

We	NN	O	O
isolated	NN	O	O
a	NN	O	O
novel	NN	O	O
subline	NN	O	O
,	NN	O	O
UT-7/GMT	NN	O	B-cell_line
,	NN	O	O
from	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
UT-7/GM	NN	O	I-cell_line
(	NN	O	O
N.	NN	O	O
Komatsu	NN	O	O
,	NN	O	O
et	NN	O	O
al.	NN	O	O
,	NN	O	O
Blood	NN	O	O
,	NN	O	O
89	NN	O	O
:	NN	O	O
4021-4033	NN	O	O
,	NN	O	O
1997	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
small	NN	O	O
population	NN	O	O
of	NN	O	O
UT-7/GM	NN	O	B-cell_line
cells	NN	O	I-cell_line
positively	NN	O	O
stained	NN	O	O
for	NN	O	O
hemoglobin	NN	O	B-protein
(	NN	O	O
Hb	NN	O	B-protein
)	NN	O	O
after	NN	O	O
a	NN	O	O
7-day	NN	O	O
exposure	NN	O	O
to	NN	O	O
TPO	NN	O	B-protein
.	NN	O	O

More	NN	O	O
than	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
TPO-treated	NN	O	B-cell_line
UT-7/GMT	NN	O	I-cell_line
cells	NN	O	I-cell_line
positively	NN	O	O
stained	NN	O	O
for	NN	O	O
Hb	NN	O	B-protein
.	NN	O	O

Using	NN	O	O
UT-7/GMT	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
examined	NN	O	O
how	NN	O	O
TPO	NN	O	B-protein
promotes	NN	O	O
hemoglobinization	NN	O	O
.	NN	O	O

TPO	NN	O	B-protein
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
TPO	NN	O	B-protein
receptor	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
erythropoietin	NN	O	B-protein
(	NN	O	I-protein
EPO	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
competition	NN	O	O
between	NN	O	O
TPO	NN	O	B-protein
and	NN	O	O
EPO	NN	O	B-protein
for	NN	O	O
binding	NN	O	O
to	NN	O	O
EPO	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
TPO	NN	O	B-protein
has	NN	O	O
a	NN	O	O
direct	NN	O	O
effect	NN	O	O
on	NN	O	O
hemoglobinization	NN	O	O
via	NN	O	O
a	NN	O	O
specific	NN	O	O
receptor	NN	O	O
on	NN	O	O
UT-7/GMT	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Isoelectric	NN	O	O
focusing	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
TPO	NN	O	B-protein
induced	NN	O	O
fetal	NN	O	O
and	NN	O	O
adult	NN	O	O
Hb	NN	O	O
synthesis	NN	O	O
,	NN	O	O
whereas	NN	O	O
EPO	NN	O	B-protein
induced	NN	O	O
embryonic	NN	O	O
,	NN	O	O
fetal	NN	O	O
,	NN	O	O
and	NN	O	O
adult	NN	O	O
Hb	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
TPO	NN	O	B-protein
has	NN	O	O
a	NN	O	O
distinct	NN	O	O
action	NN	O	O
on	NN	O	O
erythropoiesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Role	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
in	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
definitive	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
the	NN	O	O
contributions	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
to	NN	O	O
definitive	NN	O	O
hematopoiesis	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
adult	NN	O	O
mice	NN	O	O
that	NN	O	O
were	NN	O	O
rendered	NN	O	O
genetically	NN	O	O
defective	NN	O	O
in	NN	O	O
GATA-1	NN	O	B-protein
synthesis	NN	O	O
(	NN	O	O
Takahashi	NN	O	O
et	NN	O	O
al	NN	O	O
,	NN	O	O
J	NN	O	O
Biol	NN	O	O
Chem	NN	O	O
272	NN	O	O
:	NN	O	O
12611	NN	O	O
,	NN	O	O
1997	NN	O	O
)	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
GATA-1	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
located	NN	O	O
on	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
chromosome	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
randomly	NN	O	O
inactivated	NN	O	O
in	NN	O	O
every	NN	O	O
cell	NN	O	O
,	NN	O	O
heterozygous	NN	O	O
females	NN	O	O
can	NN	O	O
bear	NN	O	O
either	NN	O	O
an	NN	O	O
active	NN	O	O
wild-type	NN	O	O
or	NN	O	O
mutant	NN	O	O
(	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
GATA-1.05	NN	O	B-DNA
)	NN	O	O
GATA-1	NN	O	B-protein
allele	NN	O	O
,	NN	O	O
consequently	NN	O	O
leading	NN	O	O
to	NN	O	O
variable	NN	O	O
anemic	NN	O	O
severity	NN	O	O
.	NN	O	O

These	NN	O	O
heterozygous	NN	O	O
mutant	NN	O	O
mice	NN	O	O
usually	NN	O	O
developed	NN	O	O
normally	NN	O	O
,	NN	O	O
but	NN	O	O
they	NN	O	O
began	NN	O	O
to	NN	O	O
die	NN	O	O
after	NN	O	O
5	NN	O	O
months	NN	O	O
.	NN	O	O

These	NN	O	O
affected	NN	O	O
animals	NN	O	O
displayed	NN	O	O
marked	NN	O	O
splenomegaly	NN	O	O
,	NN	O	O
anemia	NN	O	O
,	NN	O	O
and	NN	O	O
thrombocytopenia	NN	O	O
.	NN	O	O

Proerythroblasts	NN	O	B-cell_type
and	NN	O	O
megakaryocytes	NN	O	B-cell_type
massively	NN	O	O
accumulated	NN	O	O
in	NN	O	O
the	NN	O	O
spleens	NN	O	O
of	NN	O	O
the	NN	O	O
heterozygotes	NN	O	O
,	NN	O	O
and	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
neomycin	NN	O	B-DNA
resistance	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
which	NN	O	O
is	NN	O	O
the	NN	O	O
positive	NN	O	O
selection	NN	O	O
marker	NN	O	O
in	NN	O	O
ES	NN	O	B-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
was	NN	O	O
expressed	NN	O	O
profusely	NN	O	O
in	NN	O	O
the	NN	O	O
abnormally	NN	O	O
abundant	NN	O	O
cells	NN	O	O
generated	NN	O	O
in	NN	O	O
the	NN	O	O
GATA-1.05	NN	O	B-protein
mutant	NN	O	O
females	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
observed	NN	O	O
hematopoiesis	NN	O	O
outside	NN	O	O
of	NN	O	O
the	NN	O	O
bone	NN	O	O
marrow	NN	O	O
in	NN	O	O
the	NN	O	O
affected	NN	O	O
mutant	NN	O	O
mice	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
a	NN	O	O
small	NN	O	O
number	NN	O	O
of	NN	O	O
GATA-1.05	NN	O	B-cell_line
mutant	NN	O	I-cell_line
hematopoietic	NN	O	I-cell_line
progenitor	NN	O	I-cell_line
cells	NN	O	I-cell_line
begin	NN	O	O
to	NN	O	O
proliferate	NN	O	O
vigorously	NN	O	O
during	NN	O	O
early	NN	O	O
adulthood	NN	O	O
,	NN	O	O
but	NN	O	O
because	NN	O	O
the	NN	O	O
cells	NN	O	O
are	NN	O	O
unable	NN	O	O
to	NN	O	O
terminally	NN	O	O
differentiate	NN	O	O
,	NN	O	O
this	NN	O	O
leads	NN	O	O
to	NN	O	O
progenitor	NN	O	O
proliferation	NN	O	O
in	NN	O	O
the	NN	O	O
spleen	NN	O	O
and	NN	O	O
consequently	NN	O	O
death	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
GATA-1	NN	O	B-protein
plays	NN	O	O
important	NN	O	O
in	NN	O	O
vivo	NN	O	O
roles	NN	O	O
for	NN	O	O
directing	NN	O	O
definitive	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
to	NN	O	O
differentiate	NN	O	O
along	NN	O	O
both	NN	O	O
the	NN	O	O
erythroid	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
pathways	NN	O	O
.	NN	O	O

The	NN	O	O
GATA-1	NN	O	B-protein
heterozygous	NN	O	O
mutant	NN	O	O
mouse	NN	O	O
shows	NN	O	O
a	NN	O	O
phenotype	NN	O	O
that	NN	O	O
is	NN	O	O
analogous	NN	O	O
to	NN	O	O
human	NN	O	O
myelodysplastic	NN	O	O
syndrome	NN	O	O
and	NN	O	O
thus	NN	O	O
may	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
useful	NN	O	O
model	NN	O	O
for	NN	O	O
this	NN	O	O
disorder	NN	O	O
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
-STAT	NN	O	B-protein
pathway	NN	O	O
and	NN	O	O
biologic	NN	O	O
function	NN	O	O
of	NN	O	O
IL-13	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
NK	NN	O	I-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
a	NN	O	O
comparative	NN	O	O
study	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

IL-13	NN	O	B-protein
,	NN	O	O
a	NN	O	O
cytokine	NN	O	O
similar	NN	O	O
to	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
is	NN	O	O
a	NN	O	O
regulator	NN	O	O
of	NN	O	O
human	NN	O	O
B	NN	O	O
cell	NN	O	O
and	NN	O	O
monocyte	NN	O	O
functions	NN	O	O
.	NN	O	O

Biologic	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-13	NN	O	B-protein
on	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
NK	NN	O	I-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
have	NN	O	O
not	NN	O	O
been	NN	O	O
well	NN	O	O
defined	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
may	NN	O	O
induce	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

When	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
costimulated	NN	O	O
with	NN	O	O
IL-13	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
IL-13	NN	O	B-protein
generally	NN	O	O
resulted	NN	O	O
in	NN	O	O
two	NN	O	O
types	NN	O	O
of	NN	O	O
reactivity	NN	O	O
:	NN	O	O
IL-13	NN	O	B-protein
synergized	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
to	NN	O	O
stimulate	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
or	NN	O	O
it	NN	O	O
modestly	NN	O	O
inhibited	NN	O	O
IL-2	NN	O	B-protein
-mediated	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
types	NN	O	O
of	NN	O	O
donors	NN	O	O
,	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
IL-13	NN	O	B-protein
on	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
was	NN	O	O
in	NN	O	O
marked	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
strong	NN	O	O
inhibition	NN	O	O
seen	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
in	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
IL-13	NN	O	B-protein
suppresses	NN	O	O
IL-2-induced	NN	O	O
NK	NN	O	O
cytolytic	NN	O	O
and	NN	O	O
proliferative	NN	O	O
activities	NN	O	O
although	NN	O	O
less	NN	O	O
efficiently	NN	O	O
than	NN	O	O
IL-4	NN	O	B-protein
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
IL-13	NN	O	B-protein
inhibits	NN	O	O
anti-CD3	NN	O	O
mAb/IL-2-	NN	O	O
or	NN	O	O
PHA-mediated	NN	O	O
IFN-gamma	NN	O	O
production	NN	O	O
and	NN	O	O
enhances	NN	O	O
cytolytic	NN	O	O
potential	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
IL-13	NN	O	B-protein
,	NN	O	O
like	NN	O	O
IL-4	NN	O	B-protein
,	NN	O	O
induces	NN	O	O
distinct	NN	O	O
STAT6-DNA	NN	O	B-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
and	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
and	NN	O	O
Janus	NN	O	B-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
JAK3	NN	O	B-protein
)	NN	O	O
in	NN	O	O
NK	NN	O	O
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
observed	NN	O	O
that	NN	O	O
Abs	NN	O	O
directed	NN	O	O
against	NN	O	O
unique	NN	O	O
domains	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
have	NN	O	O
differential	NN	O	O
effects	NN	O	O
on	NN	O	O
complexes	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
different	NN	O	O
STAT6	NN	O	B-protein
isoforms	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
show	NN	O	O
that	NN	O	O
IL-13	NN	O	B-protein
and	NN	O	O
IL-4	NN	O	B-protein
have	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
regulate	NN	O	O
NK	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
that	NN	O	O
IL-13	NN	O	B-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
regulator	NN	O	O
of	NN	O	O
STAT6	NN	O	B-protein
and	NN	O	O
JAK3	NN	O	B-protein
in	NN	O	O
these	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
human	NN	O	O
toll	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
:	NN	O	O
divergence	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
and	NN	O	O
JNK	NN	O	B-protein
/SAPK	NN	O	B-protein
activation	NN	O	O
upstream	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-associated	NN	O	I-protein
factor	NN	O	I-protein
6	NN	O	I-protein
(	NN	O	O
TRAF6	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
homologue	NN	O	O
of	NN	O	O
Drosophila	NN	O	B-protein
Toll	NN	O	I-protein
(	NN	O	O
hToll	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
recently	NN	O	O
cloned	NN	O	O
receptor	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-protein
1	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
IL-1R	NN	O	I-protein
)	NN	O	I-protein
superfamily	NN	O	I-protein
,	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
adaptive	NN	O	O
immunity	NN	O	O
.	NN	O	O

Signaling	NN	O	O
by	NN	O	O
hToll	NN	O	B-protein
is	NN	O	O
shown	NN	O	O
to	NN	O	O
occur	NN	O	O
through	NN	O	O
sequential	NN	O	O
recruitment	NN	O	O
of	NN	O	O
the	NN	O	O
adapter	NN	O	B-protein
molecule	NN	O	I-protein
MyD88	NN	O	B-protein
and	NN	O	O
the	NN	O	O
IL-1R-associated	NN	O	B-protein
kinase	NN	O	I-protein
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor-activated	NN	O	I-protein
factor	NN	O	I-protein
6	NN	O	I-protein
(	NN	O	O
TRAF6	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	I-protein
NF-kappaB	NN	O	I-protein
)	NN	O	I-protein
-inducing	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
NIK	NN	O	B-protein
)	NN	O	O
are	NN	O	O
both	NN	O	O
involved	NN	O	O
in	NN	O	O
subsequent	NN	O	O
steps	NN	O	O
of	NN	O	O
NF-kappaB	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
version	NN	O	O
of	NN	O	O
TRAF6	NN	O	B-protein
failed	NN	O	O
to	NN	O	O
block	NN	O	O
hToll	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
stress-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase/c-Jun	NN	O	I-protein
NH2-terminal	NN	O	I-protein
kinases	NN	O	I-protein
,	NN	O	O
thus	NN	O	O
suggesting	NN	O	O
an	NN	O	O
early	NN	O	O
divergence	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
activation	NN	O	O
attenuates	NN	O	O
apoptosis	NN	O	O
resistance	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Death-inducing	NN	O	B-protein
ligands	NN	O	I-protein
(	NN	O	O
DILs	NN	O	B-protein
)	NN	O	O
such	NN	O	O
as	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNFalpha	NN	O	B-protein
)	NN	O	O
or	NN	O	O
the	NN	O	O
cytotoxic	NN	O	O
drug	NN	O	O
doxorubicin	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
activate	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappaB	NN	O	I-protein
(	NN	O	O
NFkappaB	NN	O	B-protein
)	NN	O	O
-dependent	NN	O	O
program	NN	O	O
that	NN	O	O
may	NN	O	O
rescue	NN	O	O
cells	NN	O	O
from	NN	O	O
apoptosis	NN	O	O
induction	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
here	NN	O	O
that	NN	O	O
TRAIL	NN	O	B-protein
(	NN	O	O
TNF-related	NN	O	B-protein
apoptosis-inducing	NN	O	I-protein
ligand	NN	O	I-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
recently	NN	O	O
identified	NN	O	O
DIL	NN	O	B-protein
,	NN	O	O
also	NN	O	O
activates	NN	O	O
NFkappaB	NN	O	B-protein
in	NN	O	O
lymphoid	NN	O	O
cell	NN	O	O
lines	NN	O	O
in	NN	O	O
a	NN	O	O
kinetic	NN	O	O
similar	NN	O	O
to	NN	O	O
TNFalpha	NN	O	B-protein
.	NN	O	O

NFkappaB	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
independent	NN	O	O
from	NN	O	O
FADD	NN	O	B-protein
,	NN	O	O
caspases	NN	O	B-protein
,	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
induction	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
activity	NN	O	O
on	NN	O	O
apoptosis	NN	O	O
mediated	NN	O	O
by	NN	O	O
TRAIL	NN	O	B-protein
,	NN	O	O
CD95	NN	O	O
,	NN	O	O
TNFalpha	NN	O	B-protein
,	NN	O	O
or	NN	O	O
doxorubicin	NN	O	O
,	NN	O	O
NFkappaB	NN	O	B-protein
activation	NN	O	O
was	NN	O	O
inhibited	NN	O	O
using	NN	O	O
the	NN	O	O
proteasome	NN	O	B-protein
inhibitor	NN	O	O
N-acetyl-L-leucinyl-L-leucinyl-L-norleucinal	NN	O	O
or	NN	O	O
transient	NN	O	O
overexpression	NN	O	O
of	NN	O	O
mutant	NN	O	B-protein
IkappaBalpha	NN	O	I-protein
.	NN	O	O

Sensitivity	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
was	NN	O	O
markedly	NN	O	O
increased	NN	O	O
by	NN	O	O
these	NN	O	O
treatments	NN	O	O
in	NN	O	O
apoptosis	NN	O	O
sensitive	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
both	NN	O	O
in	NN	O	O
cell	NN	O	O
lines	NN	O	O
and	NN	O	O
in	NN	O	O
primary	NN	O	O
leukemia	NN	O	O
cells	NN	O	O
that	NN	O	O
are	NN	O	O
resistant	NN	O	O
towards	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
by	NN	O	O
DILs	NN	O	B-protein
and	NN	O	O
doxorubicin	NN	O	O
,	NN	O	O
antagonization	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
activity	NN	O	O
partially	NN	O	O
restored	NN	O	O
apoptosis	NN	O	O
sensitivity	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NFkappaB	NN	O	B-protein
activation	NN	O	O
may	NN	O	O
provide	NN	O	O
a	NN	O	O
molecular	NN	O	O
approach	NN	O	O
to	NN	O	O
increase	NN	O	O
apoptosis	NN	O	O
sensitivity	NN	O	O
in	NN	O	O
anticancer	NN	O	O
treatment	NN	O	O
.	NN	O	O

-DOCSTART-	O

Loss-	NN	O	O
and	NN	O	O
gain-of-function	NN	O	O
mutations	NN	O	O
reveal	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
of	NN	O	O
BSAP	NN	O	B-protein
(	NN	O	O
Pax-5	NN	O	B-protein
)	NN	O	O
at	NN	O	O
the	NN	O	O
start	NN	O	O
and	NN	O	O
end	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Pax-5	NN	O	B-protein
codes	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
BSAP	NN	O	B-protein
which	NN	O	O
is	NN	O	O
expressed	NN	O	O
throughout	NN	O	O
B	NN	O	O
cell	NN	O	O
development	NN	O	O
except	NN	O	O
in	NN	O	O
terminally	NN	O	B-cell_type
differentiated	NN	O	I-cell_type
plasma	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Gene	NN	O	O
targeting	NN	O	O
experiments	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
revealed	NN	O	O
a	NN	O	O
differential	NN	O	O
dependency	NN	O	O
of	NN	O	O
fetal	NN	O	O
and	NN	O	O
adult	NN	O	O
B-lymphopoiesis	NN	O	O
on	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

BSAP	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
B-lineage	NN	O	O
commitment	NN	O	O
in	NN	O	O
the	NN	O	O
fetal	NN	O	O
liver	NN	O	O
and	NN	O	O
for	NN	O	O
progression	NN	O	O
beyond	NN	O	O
an	NN	O	O
early	NN	O	O
pro-B	NN	O	O
cell	NN	O	O
stage	NN	O	O
in	NN	O	O
adult	NN	O	O
bone	NN	O	O
marrow	NN	O	O
.	NN	O	O

The	NN	O	O
characterization	NN	O	O
of	NN	O	O
Pax-5-deficient	NN	O	B-cell_type
pro-B	NN	O	I-cell_type
cells	NN	O	I-cell_type
demonstrated	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
of	NN	O	O
BSAP	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
CD19	NN	O	B-protein
,	NN	O	O
mb-1	NN	O	B-protein
(	NN	O	O
Ig	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
and	NN	O	O
N-myc	NN	O	B-DNA
genes	NN	O	I-DNA
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
developmental	NN	O	O
pathway	NN	O	O
controlling	NN	O	O
VH-to-DHJH	NN	O	O
recombination	NN	O	O
at	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-protein
heavy-chain	NN	O	I-protein
(	NN	O	I-protein
IgH	NN	O	I-protein
)	NN	O	I-protein
locus	NN	O	I-protein
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
PAX-5	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
recently	NN	O	O
shown	NN	O	O
to	NN	O	O
participate	NN	O	O
together	NN	O	O
with	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
locus	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
chromosomal	NN	O	O
translocation	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
9	NN	O	I-DNA
;	NN	O	I-DNA
14	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
p13	NN	O	I-DNA
;	NN	O	I-DNA
q32	NN	O	I-DNA
)	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
translocation	NN	O	O
is	NN	O	O
characteristic	NN	O	O
of	NN	O	O
a	NN	O	O
small	NN	O	O
subset	NN	O	O
of	NN	O	O
non-Hodgkin	NN	O	O
lymphomas	NN	O	O
exhibiting	NN	O	O
plasmacytoid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
translocated	NN	O	B-DNA
PAX-5	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
deregulated	NN	O	O
by	NN	O	O
the	NN	O	O
insertion	NN	O	O
of	NN	O	O
IgH	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
into	NN	O	O
its	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
region	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
tumorigenesis	NN	O	O
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
the	NN	O	O
shut-down	NN	O	O
of	NN	O	O
PAX-5	NN	O	B-protein
transcription	NN	O	O
and	NN	O	O
thus	NN	O	O
with	NN	O	O
the	NN	O	O
completion	NN	O	O
of	NN	O	O
plasma	NN	O	B-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O

-DOCSTART-	O

Acetylation	NN	O	O
and	NN	O	O
modulation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-protein
Kruppel-like	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EKLF	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
by	NN	O	O
interaction	NN	O	O
with	NN	O	O
histone	NN	O	B-protein
acetyltransferases	NN	O	I-protein
.	NN	O	O

Erythroid	NN	O	B-protein
Kruppel-like	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EKLF	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
red	NN	O	B-protein
cell-specific	NN	O	I-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
that	NN	O	O
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
consolidating	NN	O	O
the	NN	O	O
switch	NN	O	O
to	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
adult	NN	O	O
beta-globin	NN	O	B-protein
expression	NN	O	O
during	NN	O	O
erythroid	NN	O	O
ontogeny	NN	O	O
.	NN	O	O

EKLF	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
integrity	NN	O	O
of	NN	O	O
the	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
at	NN	O	O
the	NN	O	O
beta-like	NN	O	B-DNA
globin	NN	O	I-DNA
locus	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
it	NN	O	O
interacts	NN	O	O
with	NN	O	O
a	NN	O	O
positive-acting	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
EKLF	NN	O	B-protein
is	NN	O	O
an	NN	O	O
acetylated	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
it	NN	O	O
interacts	NN	O	O
in	NN	O	O
vivo	NN	O	O
with	NN	O	O
CBP	NN	O	B-protein
,	NN	O	O
p300	NN	O	B-protein
,	NN	O	O
and	NN	O	O
P/CAF	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
its	NN	O	O
interactions	NN	O	O
with	NN	O	O
these	NN	O	O
histone	NN	O	B-protein
acetyltransferases	NN	O	I-protein
are	NN	O	O
not	NN	O	O
equivalent	NN	O	O
,	NN	O	O
as	NN	O	O
CBP	NN	O	B-protein
and	NN	O	O
p300	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
P/CAF	NN	O	B-protein
,	NN	O	O
utilize	NN	O	O
EKLF	NN	O	B-protein
as	NN	O	O
a	NN	O	O
substrate	NN	O	O
for	NN	O	O
in	NN	O	O
vitro	NN	O	O
acetylation	NN	O	O
within	NN	O	O
its	NN	O	O
trans-	NN	O	O
activation	NN	O	B-protein
region	NN	O	I-protein
.	NN	O	O

The	NN	O	O
functional	NN	O	O
effects	NN	O	O
of	NN	O	O
these	NN	O	O
interactions	NN	O	O
are	NN	O	O
that	NN	O	O
CBP	NN	O	B-protein
and	NN	O	O
p300	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
P/CAF	NN	O	B-protein
,	NN	O	O
enhance	NN	O	O
EKLF	NN	O	B-protein
's	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
establish	NN	O	O
EKLF	NN	O	B-protein
as	NN	O	O
a	NN	O	O
tissue-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
undergoes	NN	O	O
post-translational	NN	O	O
acetylation	NN	O	O
and	NN	O	O
suggest	NN	O	O
a	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
EKLF	NN	O	B-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
alter	NN	O	O
chromatin	NN	O	B-DNA
structure	NN	O	O
and	NN	O	O
induce	NN	O	O
beta-globin	NN	O	B-protein
expression	NN	O	O
within	NN	O	O
the	NN	O	O
beta-like	NN	O	B-DNA
globin	NN	O	I-DNA
cluster	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Recognition	NN	O	O
of	NN	O	O
herpes	NN	O	B-protein
simplex	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
2	NN	O	I-protein
tegument	NN	O	I-protein
proteins	NN	O	I-protein
by	NN	O	O
CD4	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
infiltrating	NN	O	O
human	NN	O	O
genital	NN	O	O
herpes	NN	O	O
lesions	NN	O	O
.	NN	O	O

The	NN	O	O
local	NN	O	O
cellular	NN	O	O
immune	NN	O	O
response	NN	O	O
to	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
virus	NN	O	O
(	NN	O	O
HSV	NN	O	O
)	NN	O	O
is	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
recurrent	NN	O	O
HSV	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
antiviral	NN	O	O
functions	NN	O	O
of	NN	O	O
infiltrating	NN	O	B-cell_type
CD4-bearing	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
include	NN	O	O
cytotoxicity	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
viral	NN	O	O
growth	NN	O	O
,	NN	O	O
lymphokine	NN	O	B-protein
secretion	NN	O	O
,	NN	O	O
and	NN	O	O
support	NN	O	O
of	NN	O	O
humoral	NN	O	O
and	NN	O	O
CD8	NN	O	O
responses	NN	O	O
.	NN	O	O

The	NN	O	O
antigens	NN	O	O
recognized	NN	O	O
by	NN	O	O
many	NN	O	O
HSV-specific	NN	O	B-cell_type
CD4	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
localizing	NN	O	O
to	NN	O	O
genital	NN	O	O
HSV-2	NN	O	O
lesions	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
recognizing	NN	O	O
antigens	NN	O	O
encoded	NN	O	O
within	NN	O	O
map	NN	O	O
units	NN	O	O
0.67	NN	O	O
to	NN	O	O
0.73	NN	O	O
of	NN	O	O
HSV	NN	O	B-DNA
DNA	NN	O	I-DNA
are	NN	O	O
frequently	NN	O	O
recovered	NN	O	O
from	NN	O	O
herpetic	NN	O	O
lesions	NN	O	O
.	NN	O	O

Expression	NN	O	O
cloning	NN	O	O
with	NN	O	O
this	NN	O	O
region	NN	O	O
of	NN	O	O
DNA	NN	O	O
now	NN	O	O
shows	NN	O	O
that	NN	O	O
tegument	NN	O	B-protein
protein	NN	O	I-protein
VP22	NN	O	B-protein
and	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
dUTPase	NN	O	I-protein
,	NN	O	O
encoded	NN	O	O
by	NN	O	O
genes	NN	O	B-DNA
UL49	NN	O	I-DNA
and	NN	O	I-DNA
UL50	NN	O	I-DNA
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
are	NN	O	O
T-cell	NN	O	B-protein
antigens	NN	O	I-protein
.	NN	O	O

Separate	NN	O	O
epitopes	NN	O	B-protein
in	NN	O	O
VP22	NN	O	B-protein
were	NN	O	O
defined	NN	O	O
for	NN	O	O
T-cell	NN	O	B-cell_line
clones	NN	O	I-cell_line
from	NN	O	O
each	NN	O	O
of	NN	O	O
three	NN	O	O
patients	NN	O	O
.	NN	O	O

Reactivity	NN	O	O
with	NN	O	O
the	NN	O	O
tegument	NN	O	B-protein
protein	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
UL21	NN	O	B-DNA
was	NN	O	O
identified	NN	O	O
for	NN	O	O
an	NN	O	O
additional	NN	O	O
patient	NN	O	O
.	NN	O	O

Three	NN	O	O
new	NN	O	O
epitopes	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
VP16	NN	O	B-protein
,	NN	O	O
a	NN	O	O
tegument	NN	O	B-protein
protein	NN	O	I-protein
associated	NN	O	O
with	NN	O	O
VP22	NN	O	B-protein
.	NN	O	O

Some	NN	O	O
tegument-specific	NN	O	B-cell_line
CD4	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
exhibited	NN	O	O
cytotoxic	NN	O	O
activity	NN	O	O
against	NN	O	O
HSV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
herpes	NN	O	B-protein
simplex	NN	O	I-protein
tegument	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
processed	NN	O	O
for	NN	O	O
antigen	NN	O	O
presentation	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
are	NN	O	O
possible	NN	O	O
candidate	NN	O	O
compounds	NN	O	O
for	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
vaccines	NN	O	O
.	NN	O	O

-DOCSTART-	O

Fibrinogen	NN	O	B-protein
activates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
.	NN	O	O

Adhesion	NN	O	O
to	NN	O	O
extracellular	NN	O	O
matrices	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
modulate	NN	O	O
leukocyte	NN	O	O
activation	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
are	NN	O	O
not	NN	O	O
fully	NN	O	O
understood	NN	O	O
.	NN	O	O

Mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
are	NN	O	O
exposed	NN	O	O
to	NN	O	O
fibrinous	NN	O	O
provisional	NN	O	O
matrix	NN	O	O
throughout	NN	O	O
migration	NN	O	O
into	NN	O	O
inflammatory	NN	O	O
foci	NN	O	O
,	NN	O	O
so	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
undertaken	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
fibrinogen	NN	O	O
triggers	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
differentiated	NN	O	O
with	NN	O	O
PMA	NN	O	O
in	NN	O	O
nonadherent	NN	O	B-cell_line
culture	NN	O	I-cell_line
were	NN	O	O
shown	NN	O	O
to	NN	O	O
express	NN	O	O
two	NN	O	O
fibrinogen-binding	NN	O	B-protein
integrins	NN	O	I-protein
,	NN	O	O
predominately	NN	O	O
CD11b/CD18	NN	O	B-protein
,	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
CD11c/CD18	NN	O	B-protein
.	NN	O	O

Cells	NN	O	O
stimulated	NN	O	O
with	NN	O	O
fibrinogen	NN	O	B-protein
(	NN	O	O
10-100	NN	O	O
microg/ml	NN	O	O
)	NN	O	O
/Mn2+	NN	O	O
(	NN	O	O
50	NN	O	O
microM	NN	O	O
)	NN	O	O
for	NN	O	O
2	NN	O	O
h	NN	O	O
were	NN	O	O
examined	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
minimal	NN	O	O
in	NN	O	O
unstimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
was	NN	O	O
substantially	NN	O	O
up-regulated	NN	O	O
by	NN	O	O
fibrinogen	NN	O	B-protein
.	NN	O	O

Fibrinogen	NN	O	B-protein
also	NN	O	O
caused	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
SP1	NN	O	B-protein
or	NN	O	O
cAMP	NN	O	B-protein
response	NN	O	I-protein
element-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	I-protein
CREB	NN	O	I-protein
)	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Blocking	NN	O	O
mAbs	NN	O	B-protein
against	NN	O	O
CD18	NN	O	B-protein
and	NN	O	O
CD11b	NN	O	B-protein
abrogated	NN	O	O
fibrinogen	NN	O	B-protein
-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
effects	NN	O	O
on	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
,	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
plasmid	NN	O	O
containing	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
(	NN	O	O
bearing	NN	O	O
two	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
)	NN	O	O
coupled	NN	O	O
to	NN	O	O
a	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
reporter	NN	O	I-DNA
.	NN	O	O

Cells	NN	O	O
were	NN	O	O
subsequently	NN	O	O
stimulated	NN	O	O
with	NN	O	O
1	NN	O	O
)	NN	O	O
PMA	NN	O	O
for	NN	O	O
24	NN	O	O
h	NN	O	O
,	NN	O	O
inducing	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
2.6-fold	NN	O	O
,	NN	O	O
2	NN	O	O
)	NN	O	O
fibrinogen	NN	O	B-protein
/Mn2+	NN	O	O
for	NN	O	O
2	NN	O	O
h	NN	O	O
,	NN	O	O
inducing	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
3.2-fold	NN	O	O
,	NN	O	O
or	NN	O	O
3	NN	O	O
)	NN	O	O
costimulation	NN	O	O
with	NN	O	O
fibrinogen	NN	O	B-protein
and	NN	O	O
PMA	NN	O	O
,	NN	O	O
inducing	NN	O	O
5.7-fold	NN	O	O
the	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
induced	NN	O	O
by	NN	O	O
PMA	NN	O	O
alone	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
contact	NN	O	O
with	NN	O	O
fibrinogen-derived	NN	O	B-protein
proteins	NN	O	I-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
mononuclear	NN	O	O
phagocyte	NN	O	O
activation	NN	O	O
by	NN	O	O
signaling	NN	O	O
through	NN	O	O
CD11b/CD18	NN	O	B-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
selective	NN	O	O
activation	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
including	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
lupus	NN	O	O
express	NN	O	O
estrogen	NN	O	O
receptor	NN	O	O
transcripts	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
To	NN	O	O
identify	NN	O	O
and	NN	O	O
characterize	NN	O	O
estrogen	NN	O	B-RNA
receptor	NN	O	I-RNA
(	NN	O	I-RNA
ER	NN	O	I-RNA
)	NN	O	I-RNA
transcripts	NN	O	I-RNA
expressed	NN	O	O
in	NN	O	O
immune	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
(	NN	O	O
SLE	NN	O	O
)	NN	O	O
and	NN	O	O
healthy	NN	O	O
donors	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
prepared	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
SLE	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
6	NN	O	O
)	NN	O	O
and	NN	O	O
healthy	NN	O	O
donors	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
8	NN	O	O
)	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
separated	NN	O	O
into	NN	O	O
CD4	NN	O	B-cell_type
and	NN	O	O
CD8	NN	O	B-cell_type
.	NN	O	O

Some	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
estradiol	NN	O	O
,	NN	O	O
PMA	NN	O	O
,	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
.	NN	O	O

Epstein-Barr	NN	O	O
virus-transformed	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
n	NN	O	O
=	NN	O	O
7	NN	O	O
)	NN	O	O
and	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybridomas	NN	O	I-cell_line
(	NN	O	O
n	NN	O	O
=	NN	O	O
2	NN	O	O
)	NN	O	O
established	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
SLE	NN	O	O
and	NN	O	O
a	NN	O	O
healthy	NN	O	O
individual	NN	O	O
were	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
B	NN	O	O
cell	NN	O	O
source	NN	O	O
.	NN	O	O

These	NN	O	O
cells	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
ER	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
nested	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
.	NN	O	O

Amplified	NN	O	O
cDNA	NN	O	B-DNA
were	NN	O	O
sequenced	NN	O	O
by	NN	O	O
standard	NN	O	O
methods	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
In	NN	O	O
all	NN	O	O
cells	NN	O	O
tested	NN	O	O
,	NN	O	O
ER	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
expressed	NN	O	O
without	NN	O	O
prior	NN	O	O
in	NN	O	O
vitro	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Partial	NN	O	O
sequences	NN	O	O
from	NN	O	O
exons	NN	O	B-DNA
1-8	NN	O	I-DNA
were	NN	O	O
nearly	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
published	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-RNA
ER	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

There	NN	O	O
were	NN	O	O
no	NN	O	O
notable	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
ER	NN	O	B-RNA
transcripts	NN	O	I-RNA
between	NN	O	O
patients	NN	O	O
and	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

Variant	NN	O	O
receptor	NN	O	O
transcripts	NN	O	O
lacking	NN	O	O
exon	NN	O	B-DNA
5	NN	O	I-DNA
or	NN	O	O
exon	NN	O	B-DNA
7	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
encodes	NN	O	O
the	NN	O	O
hormone	NN	O	O
binding	NN	O	O
domain	NN	O	O
,	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

Precise	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
exons	NN	O	O
suggests	NN	O	O
that	NN	O	O
they	NN	O	O
are	NN	O	O
alternatively	NN	O	O
spliced	NN	O	O
transcripts	NN	O	O
.	NN	O	O

Whether	NN	O	O
the	NN	O	O
detected	NN	O	O
transcripts	NN	O	O
are	NN	O	O
translated	NN	O	O
into	NN	O	O
functional	NN	O	O
receptor	NN	O	O
proteins	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
determined	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
stimulation	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
ER	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
variants	NN	O	O
did	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
disease	NN	O	O
activity	NN	O	O
or	NN	O	O
medication	NN	O	O
.	NN	O	O

CONCLUSION	NN	O	O
:	NN	O	O
Monocytes	NN	O	B-cell_type
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
express	NN	O	O
transcripts	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
wild	NN	O	B-protein
type	NN	O	I-protein
ER	NN	O	I-protein
and	NN	O	O
the	NN	O	O
hormone	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
variants	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

DNA	NN	O	O
damaging	NN	O	O
agents	NN	O	O
induce	NN	O	O
expression	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
and	NN	O	O
subsequent	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
via	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
DNA	NN	O	O
damage	NN	O	O
and	NN	O	O
other	NN	O	O
stresses	NN	O	O
can	NN	O	O
proceed	NN	O	O
via	NN	O	O
expression	NN	O	O
of	NN	O	O
Fas	NN	O	B-protein
ligand	NN	O	I-protein
(	NN	O	O
FasL	NN	O	B-protein
)	NN	O	O
and	NN	O	O
ligation	NN	O	O
of	NN	O	O
its	NN	O	O
receptor	NN	O	O
,	NN	O	O
Fas	NN	O	O
(	NN	O	O
CD95	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
transcription	NN	O	O
factors	NN	O	O
NF-kappa	NN	O	O
B	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
crucially	NN	O	O
involved	NN	O	O
in	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
induced	NN	O	O
by	NN	O	O
etoposide	NN	O	O
,	NN	O	O
teniposide	NN	O	O
,	NN	O	O
and	NN	O	O
UV	NN	O	O
irradiation	NN	O	O
.	NN	O	O

A	NN	O	O
nondegradable	NN	O	O
mutant	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
blocked	NN	O	O
both	NN	O	O
FasL	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
induced	NN	O	O
by	NN	O	O
DNA	NN	O	O
damage	NN	O	O
but	NN	O	O
not	NN	O	O
Fas	NN	O	B-protein
ligation	NN	O	O
.	NN	O	O

These	NN	O	O
stimuli	NN	O	O
also	NN	O	O
induced	NN	O	O
the	NN	O	O
stress-activated	NN	O	O
kinase	NN	O	O
pathway	NN	O	O
(	NN	O	O
SAPK	NN	O	B-protein
/JNK	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
maximal	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

A	NN	O	O
1.2	NN	O	O
kb	NN	O	O
FasL	NN	O	B-DNA
promoter	NN	O	I-DNA
responded	NN	O	O
to	NN	O	O
DNA	NN	O	O
damage	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
coexpression	NN	O	O
with	NN	O	O
p65	NN	O	B-protein
Rel	NN	O	I-protein
or	NN	O	O
Fos/Jun	NN	O	B-protein
.	NN	O	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
relevant	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
eliminated	NN	O	O
these	NN	O	O
responses	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
AP-1	NN	O	B-protein
contributes	NN	O	O
to	NN	O	O
stress-induced	NN	O	O
apoptosis	NN	O	O
via	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
FasL	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
small	NN	O	O
,	NN	O	O
nonpeptidyl	NN	O	O
mimic	NN	O	O
of	NN	O	O
granulocyte-colony-stimulating	NN	O	B-protein
factor	NN	O	I-protein
[	NN	O	O
see	NN	O	O
commetns	NN	O	O
]	NN	O	O

A	NN	O	O
nonpeptidyl	NN	O	O
small	NN	O	O
molecule	NN	O	O
SB	NN	O	O
247464	NN	O	O
,	NN	O	O
capable	NN	O	O
of	NN	O	O
activating	NN	O	O
granulocyte-colony-stimulating	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
G-CSF	NN	O	B-protein
)	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
,	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
a	NN	O	O
high-throughput	NN	O	O
assay	NN	O	O
in	NN	O	O
cultured	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Like	NN	O	O
G-CSF	NN	O	B-protein
,	NN	O	O
SB	NN	O	O
247464	NN	O	O
induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
multiple	NN	O	B-protein
signaling	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
stimulated	NN	O	O
primary	NN	O	B-cell_type
murine	NN	O	I-cell_type
bone	NN	O	I-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
form	NN	O	O
granulocytic	NN	O	B-cell_type
colonies	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
elevated	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
neutrophil	NN	O	I-cell_type
counts	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
extracellular	NN	O	O
domain	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
G-CSF	NN	O	I-protein
receptor	NN	O	I-protein
was	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
SB	NN	O	O
247464	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
compound	NN	O	O
acts	NN	O	O
by	NN	O	O
oligomerizing	NN	O	B-protein
receptor	NN	O	I-protein
chains	NN	O	I-protein
.	NN	O	O

The	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
a	NN	O	O
small	NN	O	O
molecule	NN	O	O
can	NN	O	O
activate	NN	O	O
a	NN	O	O
receptor	NN	O	O
that	NN	O	O
normally	NN	O	O
binds	NN	O	O
a	NN	O	O
relatively	NN	O	O
large	NN	O	O
protein	NN	O	O
ligand	NN	O	O
.	NN	O	O

-DOCSTART-	O

Minimal	NN	O	O
residual	NN	O	O
disease	NN	O	O
in	NN	O	O
acute	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
with	NN	O	O
PML/RAR	NN	O	B-RNA
alpha	NN	O	I-RNA
or	NN	O	I-RNA
AML1/ETO	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
phenotypic	NN	O	O
analysis	NN	O	O
of	NN	O	O
possible	NN	O	B-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
natural	NN	O	I-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
bone	NN	O	O
marrow	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
studied	NN	O	O
minimal	NN	O	O
residual	NN	O	O
disease	NN	O	O
(	NN	O	O
MRD	NN	O	O
)	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
myeloid	NN	O	O
leukemia	NN	O	O
(	NN	O	O
AML	NN	O	O
)	NN	O	O
who	NN	O	O
have	NN	O	O
PML/RAR	NN	O	B-protein
alpha	NN	O	I-protein
or	NN	O	O
AML1/ETO	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
analysis	NN	O	O
of	NN	O	O
lymphocyte	NN	O	B-cell_type
subsets	NN	O	I-cell_type
involved	NN	O	O
in	NN	O	O
antitumor	NN	O	O
immunity	NN	O	O
.	NN	O	O

Eight	NN	O	O
patients	NN	O	O
in	NN	O	O
long-term	NN	O	O
(	NN	O	O
LT	NN	O	O
;	NN	O	O
3	NN	O	O
to	NN	O	O
15	NN	O	O
years	NN	O	O
)	NN	O	O
and	NN	O	O
15	NN	O	O
patients	NN	O	O
in	NN	O	O
short-term	NN	O	O
(	NN	O	O
ST	NN	O	O
;	NN	O	O
up	NN	O	O
to	NN	O	O
3	NN	O	O
years	NN	O	O
)	NN	O	O
remission	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
reverse	NN	O	O
transcription-polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
RT	NN	O	O
)	NN	O	O
assay	NN	O	O
,	NN	O	O
the	NN	O	O
limit	NN	O	O
of	NN	O	O
detection	NN	O	O
was	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
for	NN	O	O
PML/RAR	NN	O	B-RNA
alpha	NN	O	I-RNA
transcript	NN	O	I-RNA
and	NN	O	O
10	NN	O	O
(	NN	O	O
-4	NN	O	O
)	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
for	NN	O	O
the	NN	O	O
AML1/ETO	NN	O	B-RNA
transcript	NN	O	I-RNA
.	NN	O	O

Simultaneously	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
subsets	NN	O	I-cell_type
and	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
(	NN	O	O
PB	NN	O	O
)	NN	O	O
and	NN	O	O
bone	NN	O	O
marrow	NN	O	O
(	NN	O	O
BM	NN	O	O
)	NN	O	O
were	NN	O	O
investigated	NN	O	O
by	NN	O	O
flow	NN	O	O
cytometric	NN	O	O
analysis	NN	O	O
.	NN	O	O

Four	NN	O	O
of	NN	O	O
the	NN	O	O
eight	NN	O	O
patients	NN	O	O
in	NN	O	O
LT	NN	O	O
and	NN	O	O
7	NN	O	O
of	NN	O	O
the	NN	O	O
15	NN	O	O
patients	NN	O	O
in	NN	O	O
ST	NN	O	O
remission	NN	O	O
were	NN	O	O
MRD-positive	NN	O	O
.	NN	O	O

Although	NN	O	O
all	NN	O	O
MRD-positive	NN	O	O
patients	NN	O	O
in	NN	O	O
LT	NN	O	O
remission	NN	O	O
are	NN	O	O
still	NN	O	O
until	NN	O	O
now	NN	O	O
event-free	NN	O	O
,	NN	O	O
3	NN	O	O
of	NN	O	O
the	NN	O	O
7	NN	O	O
MRD-positive	NN	O	O
(	NN	O	O
MRD+	NN	O	O
)	NN	O	O
patients	NN	O	O
in	NN	O	O
ST	NN	O	O
remission	NN	O	O
soon	NN	O	O
relapsed	NN	O	O
.	NN	O	O

The	NN	O	O
total	NN	O	O
populations	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
+	NN	O	O
,	NN	O	O
CD8	NN	O	B-protein
+	NN	O	O
and	NN	O	O
CD56	NN	O	B-protein
+	NN	O	O
[	NN	O	O
possible	NN	O	O
T-cell	NN	O	B-cell_type
and	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	O
T/NK	NN	O	B-cell_type
)	NN	O	O
populations	NN	O	O
]	NN	O	O
in	NN	O	O
the	NN	O	O
BM	NN	O	O
of	NN	O	O
ST	NN	O	O
patients	NN	O	O
and	NN	O	O
MRD+/LT	NN	O	O
patients	NN	O	O
were	NN	O	O
significantly	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
.01	NN	O	O
)	NN	O	O
low	NN	O	O
.	NN	O	O

The	NN	O	O
CD8	NN	O	B-protein
+	NN	O	O
CD28	NN	O	B-protein
+	NN	O	O
population	NN	O	O
showed	NN	O	O
the	NN	O	O
same	NN	O	O
tendency	NN	O	O
(	NN	O	O
p	NN	O	O
<	NN	O	O
.01-.02	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
T/NK	NN	O	B-cell_type
subsets	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
BM	NN	O	O
of	NN	O	O
MRD-negative	NN	O	O
(	NN	O	O
MRD-	NN	O	O
)	NN	O	O
LT	NN	O	O
(	NN	O	O
MRD-/LT	NN	O	O
)	NN	O	O
patients	NN	O	O
showed	NN	O	O
similar	NN	O	O
numbers	NN	O	O
of	NN	O	O
cells	NN	O	O
as	NN	O	O
normal	NN	O	O
volunteers	NN	O	O
.	NN	O	O

Basically	NN	O	O
,	NN	O	O
the	NN	O	O
total	NN	O	O
percentage	NN	O	O
of	NN	O	O
the	NN	O	O
CD4+	NN	O	B-cell_type
,	NN	O	I-cell_type
CD8+	NN	O	I-cell_type
and	NN	O	I-cell_type
CD56+	NN	O	I-cell_type
cell	NN	O	I-cell_type
populations	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
BM	NN	O	O
was	NN	O	O
increased	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
following	NN	O	O
order	NN	O	O
:	NN	O	O
MRD-/LT	NN	O	O
patients	NN	O	O
,	NN	O	O
normal	NN	O	O
volunteers	NN	O	O
,	NN	O	O
MRD+/LT	NN	O	O
patients	NN	O	O
and	NN	O	O
MRD+	NN	O	O
or	NN	O	O
-/ST	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
percentages	NN	O	O
of	NN	O	O
the	NN	O	O
T/NK-cell	NN	O	B-cell_type
subsets	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
PB	NN	O	O
were	NN	O	O
not	NN	O	O
significantly	NN	O	O
different	NN	O	O
among	NN	O	O
these	NN	O	O
groups	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
difference	NN	O	O
of	NN	O	O
the	NN	O	O
possible	NN	O	O
T/NK-cell	NN	O	B-cell_type
phenotype	NN	O	O
in	NN	O	O
the	NN	O	O
BM	NN	O	O
may	NN	O	O
strongly	NN	O	O
influence	NN	O	O
clinical	NN	O	O
and	NN	O	O
molecular	NN	O	O
remission	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
still	NN	O	O
remain	NN	O	O
to	NN	O	O
be	NN	O	O
confirmed	NN	O	O
by	NN	O	O
further	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
anti-tumor	NN	O	O
immunity	NN	O	O
of	NN	O	O
T/NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
AML	NN	O	O
in	NN	O	O
remission	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mycobacterium	NN	O	O
tuberculosis	NN	O	O
mannose-capped	NN	O	O
lipoarabinomannan	NN	O	O
can	NN	O	O
induce	NN	O	O
NF-kappaB	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Tuberculosis	NN	O	O
has	NN	O	O
emerged	NN	O	O
as	NN	O	O
an	NN	O	O
epidemic	NN	O	O
,	NN	O	O
extended	NN	O	O
by	NN	O	O
the	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
individuals	NN	O	O
infected	NN	O	O
with	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
major	NN	O	O
goal	NN	O	O
of	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
mycobacterial	NN	O	B-cell_type
cell	NN	O	I-cell_type
wall	NN	O	O
component	NN	O	O
mannose-capped	NN	O	O
lipoarabinomannan	NN	O	O
(	NN	O	O
ManLAM	NN	O	O
)	NN	O	O
of	NN	O	O
Mycobacterium	NN	O	O
tuberculosis	NN	O	O
(	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
)	NN	O	O
could	NN	O	O
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
cell	NN	O	O
culture	NN	O	O
system	NN	O	O
.	NN	O	O

These	NN	O	O
experiments	NN	O	O
are	NN	O	O
of	NN	O	O
prime	NN	O	O
importance	NN	O	O
considering	NN	O	O
that	NN	O	O
CD4-expressing	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
represent	NN	O	O
the	NN	O	O
major	NN	O	O
virus	NN	O	O
reservoir	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
infected	NN	O	O
individuals	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
1G5	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
harbouring	NN	O	O
the	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
,	NN	O	O
it	NN	O	O
was	NN	O	O
first	NN	O	O
found	NN	O	O
that	NN	O	O
culture	NN	O	O
protein	NN	O	O
filtrates	NN	O	O
(	NN	O	O
CFP	NN	O	O
)	NN	O	O
from	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
or	NN	O	O
purified	NN	O	O
ManLAM	NN	O	O
could	NN	O	O
activate	NN	O	O
HIV-1	NN	O	B-DNA
LTR-dependent	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
unlike	NN	O	O
similarly	NN	O	O
prepared	NN	O	O
CFP	NN	O	O
extracts	NN	O	O
devoid	NN	O	O
of	NN	O	O
ManLAM	NN	O	O
.	NN	O	O

The	NN	O	O
implication	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	I-protein
s	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
and/or	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
was	NN	O	O
highlighted	NN	O	O
by	NN	O	O
the	NN	O	O
abrogation	NN	O	O
of	NN	O	O
the	NN	O	O
ManLAM-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-DNA
LTR-driven	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
using	NN	O	O
herbimycin	NN	O	O
A	NN	O	O
and	NN	O	O
H7	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
also	NN	O	O
determined	NN	O	O
,	NN	O	O
using	NN	O	O
electrophoresis	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
that	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
ManLAM	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappaB	NN	O	I-protein
.	NN	O	O

M.	NN	O	O
tuberculosis	NN	O	O
ManLAM	NN	O	O
resulted	NN	O	O
in	NN	O	O
clear	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
luciferase	NN	O	B-DNA
gene	NN	O	I-DNA
placed	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
wild-type	NN	O	B-DNA
,	NN	O	I-DNA
but	NN	O	I-DNA
not	NN	O	I-DNA
the	NN	O	I-DNA
kappaB-mutated	NN	O	I-DNA
,	NN	O	I-DNA
HIV-1	NN	O	I-DNA
LTR	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
ManLAM-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
transcription	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
autocrine	NN	O	O
or	NN	O	O
paracrine	NN	O	O
action	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
TNF-alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
M.	NN	O	O
tuberculosis	NN	O	O
can	NN	O	O
upregulate	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
could	NN	O	O
thus	NN	O	O
have	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
influence	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
quasispecies	NN	O	I-DNA
differ	NN	O	O
in	NN	O	O
basal	NN	O	O
transcription	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
recruitment	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
glial	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
generation	NN	O	O
of	NN	O	O
genomic	NN	O	O
diversity	NN	O	O
during	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
infection	NN	O	O
has	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
affect	NN	O	O
all	NN	O	O
aspects	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
,	NN	O	O
including	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
proviral	NN	O	B-DNA
genome	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
gain	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
impact	NN	O	O
of	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
sequence	NN	O	O
diversity	NN	O	O
on	NN	O	O
LTR-directed	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
(	NN	O	O
CNS	NN	O	O
)	NN	O	O
and	NN	O	O
immune	NN	O	O
system	NN	O	O
,	NN	O	O
we	NN	O	O
amplified	NN	O	O
and	NN	O	O
cloned	NN	O	O
LTRs	NN	O	B-DNA
from	NN	O	O
proviral	NN	O	B-DNA
DNA	NN	O	I-DNA
in	NN	O	O
HIV-1-infected	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
nineteen	NN	O	O
LTRs	NN	O	B-DNA
cloned	NN	O	O
from	NN	O	O
2	NN	O	O
adult	NN	O	O
and	NN	O	O
3	NN	O	O
pediatric	NN	O	O
patients	NN	O	O
revealed	NN	O	O
an	NN	O	O
average	NN	O	O
of	NN	O	O
33	NN	O	O
nucleotide	NN	O	O
changes	NN	O	O
(	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
LAI	NN	O	B-DNA
LTR	NN	O	I-DNA
)	NN	O	O
within	NN	O	O
the	NN	O	O
455-bp	NN	O	B-DNA
U3	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
analyses	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
neuroglial	NN	O	O
and	NN	O	O
lymphocytic	NN	O	O
origin	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
some	NN	O	O
of	NN	O	O
these	NN	O	O
LTRs	NN	O	B-DNA
had	NN	O	O
activities	NN	O	O
which	NN	O	O
varied	NN	O	O
significantly	NN	O	O
from	NN	O	O
the	NN	O	O
LAI	NN	O	B-DNA
LTR	NN	O	I-DNA
in	NN	O	O
U-373	NN	O	B-cell_line
MG	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
an	NN	O	O
astrocytoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
a	NN	O	O
CD4-positive	NN	O	B-cell_line
lymphocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
.	NN	O	O

While	NN	O	O
LTRs	NN	O	B-DNA
which	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
highest	NN	O	O
activities	NN	O	O
in	NN	O	O
U-373	NN	O	B-cell_line
MG	NN	O	I-cell_line
cells	NN	O	I-cell_line
also	NN	O	O
yielded	NN	O	O
high	NN	O	O
activities	NN	O	O
in	NN	O	O
Jurkat	NN	O	O
cells	NN	O	O
,	NN	O	O
the	NN	O	O
LTRs	NN	O	B-DNA
were	NN	O	O
generally	NN	O	O
more	NN	O	O
active	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
when	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
LAI	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

Differences	NN	O	O
in	NN	O	O
LTR	NN	O	B-DNA
sequence	NN	O	I-DNA
also	NN	O	O
resulted	NN	O	O
in	NN	O	O
differences	NN	O	O
in	NN	O	O
transcription	NN	O	O
factor	NN	O	O
recruitment	NN	O	O
to	NN	O	O
cis-acting	NN	O	B-DNA
sites	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
U3	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
,	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
sequence	NN	O	O
variation	NN	O	O
impacted	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
an	NN	O	O
activating	NN	O	B-DNA
transcription	NN	O	I-DNA
factor/cAMP	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
binding	NN	O	I-DNA
(	NN	O	I-DNA
ATF/CREB	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
located	NN	O	O
between	NN	O	O
the	NN	O	O
LEF-1	NN	O	B-DNA
and	NN	O	I-DNA
distal	NN	O	I-DNA
NF-kappaB	NN	O	I-DNA
transcription	NN	O	I-DNA
factor	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
)	NN	O	O
that	NN	O	O
we	NN	O	O
identified	NN	O	O
in	NN	O	O
previous	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
LTR	NN	O	B-DNA
sequence	NN	O	I-DNA
changes	NN	O	O
can	NN	O	O
significantly	NN	O	O
affect	NN	O	O
basal	NN	O	O
LTR	NN	O	B-DNA
function	NN	O	O
and	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
recruitment	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
,	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
alter	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
viral	NN	O	O
replication	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
CNS	NN	O	O
and	NN	O	O
immune	NN	O	O
system	NN	O	O
origin	NN	O	O
.	NN	O	O

-DOCSTART-	O

HMG	NN	O	B-protein
box	NN	O	I-protein
containing	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
lymphocyte	NN	O	B-cell_type
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
the	NN	O	O
mammalian	NN	O	B-DNA
sex-determining	NN	O	I-DNA
gene	NN	O	I-DNA
Sry	NN	O	I-DNA
has	NN	O	O
led	NN	O	O
to	NN	O	O
the	NN	O	O
discovery	NN	O	O
of	NN	O	O
a	NN	O	O
large	NN	O	O
family	NN	O	O
of	NN	O	O
related	NN	O	O
(	NN	O	O
'	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
'	NN	O	O
)	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
control	NN	O	O
developmental	NN	O	O
events	NN	O	O
in	NN	O	O
yeast	NN	O	O
,	NN	O	O
C.	NN	O	O
elegans	NN	O	O
,	NN	O	O
Drosophila	NN	O	O
and	NN	O	O
vertebrates	NN	O	O
.	NN	O	O

In	NN	O	O
lymphocyte	NN	O	B-cell_type
differentiation	NN	O	O
,	NN	O	O
several	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
proteins	NN	O	I-protein
play	NN	O	O
a	NN	O	O
decisive	NN	O	O
role	NN	O	O
.	NN	O	O

Sox-4	NN	O	B-protein
is	NN	O	O
important	NN	O	O
for	NN	O	O
very	NN	O	O
early	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
while	NN	O	O
TCF-1/LEF-1	NN	O	B-protein
play	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
early	NN	O	O
thymocyte	NN	O	O
development	NN	O	O
.	NN	O	O

TCF/LEF	NN	O	B-protein
proteins	NN	O	I-protein
have	NN	O	O
recently	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
constitute	NN	O	O
a	NN	O	O
downstream	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
Wingless/Wnt	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
.	NN	O	O

In	NN	O	O
flies	NN	O	O
,	NN	O	O
this	NN	O	O
pathway	NN	O	O
controls	NN	O	O
segment	NN	O	O
polarity	NN	O	O
;	NN	O	O
in	NN	O	O
Xenopus	NN	O	O
it	NN	O	O
controls	NN	O	O
the	NN	O	O
definition	NN	O	O
of	NN	O	O
the	NN	O	O
body	NN	O	O
axis	NN	O	O
.	NN	O	O

Deregulation	NN	O	O
of	NN	O	O
the	NN	O	O
pathway	NN	O	O
occurs	NN	O	O
in	NN	O	O
several	NN	O	O
human	NN	O	O
tumors	NN	O	O
.	NN	O	O

These	NN	O	O
insights	NN	O	O
in	NN	O	O
the	NN	O	O
molecular	NN	O	O
events	NN	O	O
that	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
TCF/LEF	NN	O	B-protein
function	NN	O	O
in	NN	O	O
these	NN	O	O
organisms	NN	O	O
may	NN	O	O
eventually	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
these	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
lymphoid	NN	O	O
development	NN	O	O

-DOCSTART-	O

Single	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
of	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
mutations	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
androgen	NN	O	O
insensitivity	NN	O	O
syndromes	NN	O	O
:	NN	O	O
application	NN	O	O
for	NN	O	O
diagnosis	NN	O	O
,	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
,	NN	O	O
and	NN	O	O
therapy	NN	O	O
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
are	NN	O	O
associated	NN	O	O
with	NN	O	O
androgen	NN	O	O
insensitivity	NN	O	O
syndromes	NN	O	O
,	NN	O	O
a	NN	O	O
heterogeneous	NN	O	O
group	NN	O	O
of	NN	O	O
related	NN	O	O
disorders	NN	O	O
involving	NN	O	O
defective	NN	O	O
sexual	NN	O	O
differentiation	NN	O	O
in	NN	O	O
karyotypic	NN	O	O
males	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
address	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
rapid	NN	O	O
mutational	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
for	NN	O	O
initial	NN	O	O
diagnosis	NN	O	O
,	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
,	NN	O	O
and	NN	O	O
molecular	NN	O	O
subclassification	NN	O	O
of	NN	O	O
affected	NN	O	O
patients	NN	O	O
and	NN	O	O
their	NN	O	O
families	NN	O	O
.	NN	O	O

DNA	NN	O	O
from	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
of	NN	O	O
six	NN	O	O
patients	NN	O	O
from	NN	O	O
five	NN	O	O
families	NN	O	O
with	NN	O	O
various	NN	O	O
degrees	NN	O	O
of	NN	O	O
androgen	NN	O	O
insensitivity	NN	O	O
was	NN	O	O
studied	NN	O	O
.	NN	O	O

Exons	NN	O	B-DNA
2	NN	O	I-DNA
to	NN	O	I-DNA
8	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
were	NN	O	O
analyzed	NN	O	O
using	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
single	NN	O	O
strand	NN	O	O
conformation	NN	O	O
polymorphism	NN	O	O
analysis	NN	O	O
and	NN	O	O
direct	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
.	NN	O	O

Female	NN	O	O
family	NN	O	O
members	NN	O	O
were	NN	O	O
also	NN	O	O
studied	NN	O	O
to	NN	O	O
identify	NN	O	O
heterozygote	NN	O	O
carriers	NN	O	O
.	NN	O	O

Point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
AR	NN	O	B-DNA
gene	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
in	NN	O	O
all	NN	O	O
six	NN	O	O
patients	NN	O	O
,	NN	O	O
and	NN	O	O
all	NN	O	O
mutations	NN	O	O
caused	NN	O	O
amino	NN	O	O
acid	NN	O	O
substitutions	NN	O	O
.	NN	O	O

One	NN	O	O
patient	NN	O	O
with	NN	O	O
incomplete	NN	O	O
androgen	NN	O	O
insensitivity	NN	O	O
was	NN	O	O
a	NN	O	O
mosaic	NN	O	O
for	NN	O	O
the	NN	O	O
mutation	NN	O	O
.	NN	O	O

Four	NN	O	O
of	NN	O	O
the	NN	O	O
five	NN	O	O
mothers	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
young	NN	O	O
sister	NN	O	O
of	NN	O	O
one	NN	O	O
patient	NN	O	O
,	NN	O	O
were	NN	O	O
carriers	NN	O	O
of	NN	O	O
the	NN	O	O
mutation	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
affected	NN	O	O
child	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
new	NN	O	O
mutations	NN	O	O
may	NN	O	O
occur	NN	O	O
in	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
leading	NN	O	O
to	NN	O	O
sporadic	NN	O	O
androgen	NN	O	O
insensitivity	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Molecular	NN	O	O
genetic	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
variant	NN	O	B-DNA
allele	NN	O	I-DNA
can	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
primary	NN	O	O
tool	NN	O	O
for	NN	O	O
diagnosis	NN	O	O
and	NN	O	O
subsequent	NN	O	O
therapy	NN	O	O
,	NN	O	O
and	NN	O	O
can	NN	O	O
provide	NN	O	O
a	NN	O	O
basis	NN	O	O
for	NN	O	O
distinguishing	NN	O	O
heterozygous	NN	O	O
carriers	NN	O	O
in	NN	O	O
familial	NN	O	O
androgen	NN	O	O
resistance	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
carriers	NN	O	O
is	NN	O	O
of	NN	O	O
substantial	NN	O	O
clinical	NN	O	O
importance	NN	O	O
for	NN	O	O
genetic	NN	O	O
counseling	NN	O	O
.	NN	O	O

-DOCSTART-	O

Proliferation	NN	O	O
index	NN	O	O
as	NN	O	O
a	NN	O	O
prognostic	NN	O	O
marker	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

BACKGROUND	NN	O	O
.	NN	O	O

The	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
of	NN	O	O
tumors	NN	O	O
has	NN	O	O
been	NN	O	O
extensively	NN	O	O
investigated	NN	O	O
with	NN	O	O
different	NN	O	O
approaches	NN	O	O
,	NN	O	O
among	NN	O	O
which	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
Ki-67	NN	O	I-protein
represents	NN	O	O
an	NN	O	O
easy	NN	O	O
and	NN	O	O
reliable	NN	O	O
means	NN	O	O
of	NN	O	O
assessing	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
the	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
of	NN	O	O
129	NN	O	O
primary	NN	O	O
breast	NN	O	O
cancers	NN	O	O
was	NN	O	O
investigated	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
results	NN	O	O
were	NN	O	O
related	NN	O	O
to	NN	O	O
prognosis	NN	O	O
.	NN	O	O

METHODS	NN	O	O
.	NN	O	O

Tumor	NN	O	O
samples	NN	O	O
,	NN	O	O
obtained	NN	O	O
from	NN	O	O
129	NN	O	O
patients	NN	O	O
who	NN	O	O
underwent	NN	O	O
surgery	NN	O	O
between	NN	O	O
January	NN	O	O
1987	NN	O	O
and	NN	O	O
December	NN	O	O
1988	NN	O	O
,	NN	O	O
were	NN	O	O
processed	NN	O	O
for	NN	O	O
staining	NN	O	O
by	NN	O	O
an	NN	O	O
immunohistochemical	NN	O	O
procedure	NN	O	O
(	NN	O	O
avidin-biotin	NN	O	B-protein
complex	NN	O	I-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
median	NN	O	O
time	NN	O	O
of	NN	O	O
observation	NN	O	O
was	NN	O	O
42	NN	O	O
months	NN	O	O
(	NN	O	O
range	NN	O	O
,	NN	O	O
31-55	NN	O	O
months	NN	O	O
)	NN	O	O
.	NN	O	O

Life-table	NN	O	O
analysis	NN	O	O
(	NN	O	O
Mantel-Cox	NN	O	O
)	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
assess	NN	O	O
the	NN	O	O
probability	NN	O	O
of	NN	O	O
disease-free	NN	O	O
survival	NN	O	O
(	NN	O	O
DFS	NN	O	O
)	NN	O	O
and	NN	O	O
overall	NN	O	O
survival	NN	O	O
(	NN	O	O
OS	NN	O	O
)	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
.	NN	O	O

Tumors	NN	O	O
with	NN	O	O
high	NN	O	O
Ki-67	NN	O	B-protein
proliferation	NN	O	O
indices	NN	O	O
(	NN	O	O
>	NN	O	O
20	NN	O	O
%	NN	O	O
)	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
higher	NN	O	O
4-year	NN	O	O
probability	NN	O	O
of	NN	O	O
relapse	NN	O	O
of	NN	O	O
disease	NN	O	O
(	NN	O	O
55.3	NN	O	O
%	NN	O	O
versus	NN	O	O
79.1	NN	O	O
%	NN	O	O
;	NN	O	O
P	NN	O	O
=	NN	O	O
0.003	NN	O	O
)	NN	O	O
and	NN	O	O
death	NN	O	O
(	NN	O	O
71	NN	O	O
%	NN	O	O
versus	NN	O	O
95.6	NN	O	O
%	NN	O	O
;	NN	O	O
P	NN	O	O
=	NN	O	O
0.00005	NN	O	O
)	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
tumors	NN	O	O
with	NN	O	O
low	NN	O	O
Ki-67	NN	O	B-protein
values	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
this	NN	O	O
proliferative	NN	O	O
parameter	NN	O	O
maintained	NN	O	O
its	NN	O	O
prognostic	NN	O	O
significance	NN	O	O
when	NN	O	O
the	NN	O	O
patients	NN	O	O
were	NN	O	O
stratified	NN	O	O
according	NN	O	O
to	NN	O	O
lymph	NN	O	O
node	NN	O	O
involvement	NN	O	O
,	NN	O	O
menopausal	NN	O	O
status	NN	O	O
,	NN	O	O
and	NN	O	O
nuclear	NN	O	B-protein
estrogen	NN	O	I-protein
receptor	NN	O	I-protein
content	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
.	NN	O	O

Tumor	NN	O	O
proliferative	NN	O	O
activity	NN	O	O
as	NN	O	O
evaluated	NN	O	O
by	NN	O	O
the	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
Ki-67	NN	O	I-protein
seems	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
effective	NN	O	O
indicator	NN	O	O
of	NN	O	O
prognosis	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
for	NN	O	O
DFS	NN	O	O
and	NN	O	O
OS	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
Ets-related	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Elf-1	NN	O	B-protein
by	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
retinoblastoma	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
(	NN	O	O
Rb	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
phosphoprotein	NN	O	I-protein
that	NN	O	O
regulates	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
.	NN	O	O

Elf-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
lymphoid-specific	NN	O	B-protein
Ets	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
regulates	NN	O	O
inducible	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
Elf-1	NN	O	B-protein
contains	NN	O	O
a	NN	O	O
sequence	NN	O	O
motif	NN	O	O
that	NN	O	O
is	NN	O	O
highly	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
Rb	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
of	NN	O	O
several	NN	O	O
viral	NN	O	O
oncoproteins	NN	O	B-protein
and	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
pocket	NN	O	O
region	NN	O	O
of	NN	O	O
Rb	NN	O	B-protein
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Elf-1	NN	O	B-protein
binds	NN	O	O
exclusively	NN	O	O
to	NN	O	O
the	NN	O	O
underphosphorylated	NN	O	O
form	NN	O	O
of	NN	O	O
Rb	NN	O	B-protein
and	NN	O	O
fails	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
Rb	NN	O	B-protein
mutants	NN	O	I-protein
derived	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
retinoblastoma	NN	O	O
.	NN	O	O

Co-immunoprecipitation	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
an	NN	O	O
association	NN	O	O
between	NN	O	O
Elf-1	NN	O	B-protein
and	NN	O	O
Rb	NN	O	B-protein
in	NN	O	O
resting	NN	O	B-cell_type
normal	NN	O	I-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

After	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Rb	NN	O	B-protein
results	NN	O	O
in	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
Elf-1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
correlated	NN	O	O
temporally	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Elf-1	NN	O	B-protein
-mediated	NN	O	O
transcription	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
a	NN	O	O
phosphorylation-defective	NN	O	O
form	NN	O	O
of	NN	O	O
Rb	NN	O	B-protein
inhibited	NN	O	O
Elf-1	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
during	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Rb	NN	O	B-protein
interacts	NN	O	O
specifically	NN	O	O
with	NN	O	O
a	NN	O	O
lineage-restricted	NN	O	B-protein
Ets	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

This	NN	O	O
regulated	NN	O	O
interaction	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
coordination	NN	O	O
of	NN	O	O
lineage-specific	NN	O	O
effector	NN	O	O
functions	NN	O	O
such	NN	O	O
as	NN	O	O
lymphokine	NN	O	B-protein
production	NN	O	O
with	NN	O	O
cell	NN	O	O
cycle	NN	O	O
progression	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-cell_line
CD3-CD16+	NN	O	I-cell_line
natural	NN	O	I-cell_line
killer	NN	O	I-cell_line
cells	NN	O	I-cell_line
express	NN	O	O
the	NN	O	O
hGATA-3	NN	O	B-protein
T	NN	O	I-protein
cell	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
an	NN	O	O
unrearranged	NN	O	O
2.3-kb	NN	O	B-protein
TcR	NN	O	I-protein
delta	NN	O	I-protein
transcript	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	I-protein
TcR	NN	O	I-protein
)	NN	O	I-protein
delta	NN	O	I-protein
transcripts	NN	O	I-protein
expressed	NN	O	O
by	NN	O	O
CD3-CD16+	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
we	NN	O	O
investigated	NN	O	O
whether	NN	O	O
these	NN	O	O
cells	NN	O	O
expressed	NN	O	O
the	NN	O	O
hGATA-3	NN	O	B-protein
T	NN	O	I-protein
cell	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
the	NN	O	O
recombination-activating	NN	O	B-DNA
gene	NN	O	I-DNA
(	NN	O	I-DNA
RAG	NN	O	I-DNA
)	NN	O	I-DNA
-1	NN	O	I-DNA
.	NN	O	O

Multiple	NN	O	O
TcR	NN	O	B-protein
delta	NN	O	I-protein
transcripts	NN	O	I-protein
deriving	NN	O	O
from	NN	O	O
an	NN	O	O
unrearranged	NN	O	O
TcR	NN	O	B-DNA
delta	NN	O	I-DNA
gene	NN	O	I-DNA
were	NN	O	O
detected	NN	O	O
in	NN	O	O
both	NN	O	O
polyclonal	NN	O	B-cell_line
and	NN	O	I-cell_line
clonal	NN	O	I-cell_line
CD3-CD16+	NN	O	I-cell_line
natural	NN	O	I-cell_line
killer	NN	O	I-cell_line
(	NN	O	I-cell_line
NK	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Two	NN	O	O
unrearranged	NN	O	O
TcR	NN	O	B-protein
delta	NN	O	I-protein
transcripts	NN	O	I-protein
had	NN	O	O
a	NN	O	O
size	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
TcR	NN	O	B-RNA
delta	NN	O	I-RNA
mRNA	NN	O	I-RNA
(	NN	O	O
2.3	NN	O	O
and	NN	O	O
1.3	NN	O	O
kb	NN	O	O
)	NN	O	O
found	NN	O	O
in	NN	O	O
TcR	NN	O	B-cell_line
gamma/delta+	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Sequence	NN	O	O
analysis	NN	O	O
of	NN	O	O
nine	NN	O	O
different	NN	O	O
2.3-kb	NN	O	O
cDNA	NN	O	O
clones	NN	O	O
obtained	NN	O	O
from	NN	O	O
NK-derived	NN	O	O
polyA+	NN	O	O
RNA	NN	O	O
confirmed	NN	O	O
that	NN	O	O
they	NN	O	O
corresponded	NN	O	O
to	NN	O	O
an	NN	O	O
unrearranged	NN	O	O
TcR	NN	O	B-DNA
delta	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
cDNA	NN	O	O
were	NN	O	O
2343	NN	O	O
bp	NN	O	O
long	NN	O	O
and	NN	O	O
their	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
site	NN	O	O
was	NN	O	O
located	NN	O	O
814	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
J	NN	O	B-DNA
delta	NN	O	I-DNA
1	NN	O	I-DNA
segment	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
sequence	NN	O	O
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
J	NN	O	B-DNA
delta	NN	O	I-DNA
1	NN	O	I-DNA
segment	NN	O	I-DNA
corresponded	NN	O	O
to	NN	O	O
the	NN	O	O
previously	NN	O	O
reported	NN	O	O
germ-line	NN	O	O
sequence	NN	O	O
.	NN	O	O

The	NN	O	O
J	NN	O	B-DNA
delta	NN	O	I-DNA
1	NN	O	I-DNA
segment	NN	O	I-DNA
was	NN	O	O
correctly	NN	O	O
spliced	NN	O	O
to	NN	O	O
C	NN	O	B-DNA
delta	NN	O	I-DNA
;	NN	O	O
in	NN	O	O
addition	NN	O	O
the	NN	O	O
four	NN	O	O
C	NN	O	O
delta	NN	O	O
exons	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
already	NN	O	O
assembled	NN	O	O
.	NN	O	O

Two	NN	O	O
polyadenylation	NN	O	O
sites	NN	O	O
were	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
fourth	NN	O	B-DNA
C	NN	O	I-DNA
delta	NN	O	I-DNA
exon	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
only	NN	O	O
that	NN	O	O
located	NN	O	O
at	NN	O	O
the	NN	O	O
3	NN	O	O
'	NN	O	O
end	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
utilized	NN	O	O
in	NN	O	O
the	NN	O	O
2.3-kb	NN	O	B-DNA
cDNA	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
hGATA-3	NN	O	B-protein
,	NN	O	O
a	NN	O	O
T	NN	O	B-protein
cell-specific	NN	O	I-protein
factor	NN	O	I-protein
known	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
TcR	NN	O	B-DNA
delta	NN	O	I-DNA
locus	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
,	NN	O	O
in	NN	O	O
cultured	NN	O	O
NK	NN	O	B-cell_line
cell	NN	O	I-cell_line
population	NN	O	I-cell_line
and	NN	O	O
clones	NN	O	O
(	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
freshly	NN	O	O
derived	NN	O	O
cell	NN	O	O
populations	NN	O	O
)	NN	O	O
.	NN	O	O

All	NN	O	O
NK	NN	O	B-cell_line
clones	NN	O	I-cell_line
and	NN	O	O
cell	NN	O	O
lines	NN	O	O
studied	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
express	NN	O	O
hGATA-3-specific	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
hGATA-3	NN	O	B-protein
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
unrearranged	NN	O	O
TcR	NN	O	B-DNA
delta	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
no	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
RAG-1	NN	O	B-DNA
gene	NN	O	I-DNA
could	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
all	NN	O	O
NK	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
or	NN	O	O
clones	NN	O	O
analyzed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Negative	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
interleukin	NN	O	I-DNA
2	NN	O	I-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
glucocorticoids	NN	O	O
through	NN	O	O
interference	NN	O	O
with	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
AP-1	NN	O	B-protein
and	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
is	NN	O	O
affected	NN	O	O
by	NN	O	O
several	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
asked	NN	O	O
whether	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
Dex	NN	O	O
)	NN	O	O
and	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
inhibit	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
IL-2	NN	O	O
promoter	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
analyzed	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
DNA	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

Both	NN	O	O
Dex	NN	O	O
and	NN	O	O
CsA	NN	O	O
inhibited	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
transcription	NN	O	O
factors	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
of	NN	O	O
NF-kB	NN	O	B-protein
and	NN	O	O
OCT-1/OAF	NN	O	B-protein
,	NN	O	O
to	NN	O	O
their	NN	O	O
corresponding	NN	O	O
sites	NN	O	O
on	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
correlate	NN	O	O
changes	NN	O	O
in	NN	O	O
nuclear	NN	O	O
factor	NN	O	O
binding	NN	O	O
in	NN	O	O
vitro	NN	O	O
with	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
define	NN	O	O
the	NN	O	O
structural	NN	O	O
requirements	NN	O	O
for	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
repression	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
transient	NN	O	O
DNA	NN	O	O
transfections	NN	O	O
.	NN	O	O

Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
transfected	NN	O	O
with	NN	O	O
plasmids	NN	O	B-DNA
containing	NN	O	O
either	NN	O	O
the	NN	O	O
intact	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
or	NN	O	O
its	NN	O	O
AP-1	NN	O	B-DNA
,	NN	O	I-DNA
NF-AT	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
NF-kB	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

Dex	NN	O	O
inhibited	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-DNA
and	NN	O	O
NF-kB	NN	O	B-DNA
plasmids	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
CsA	NN	O	O
inhibited	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
and	NN	O	O
NF-kB	NN	O	B-DNA
plasmids	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
both	NN	O	O
Dex	NN	O	O
and	NN	O	O
CsA	NN	O	O
inhibited	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
through	NN	O	O
interference	NN	O	O
with	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP-1	NN	O	B-protein
and	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
,	NN	O	O
while	NN	O	O
maximum	NN	O	O
inhibition	NN	O	O
may	NN	O	O
involve	NN	O	O
interaction	NN	O	O
with	NN	O	O
both	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
the	NN	O	O
primary	NN	O	O
target	NN	O	O
of	NN	O	O
Dex	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
mediator	NN	O	O
of	NN	O	O
lipopolysaccharide	NN	O	O
effects	NN	O	O
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
certain	NN	O	O
cell	NN	O	O
types	NN	O	O
to	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
leads	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
an	NN	O	O
inducible	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

One	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
's	NN	O	O
unique	NN	O	O
properties	NN	O	O
is	NN	O	O
its	NN	O	O
posttranslational	NN	O	O
activation	NN	O	O
via	NN	O	O
release	NN	O	O
of	NN	O	O
an	NN	O	O
inhibitory	NN	O	B-protein
subunit	NN	O	I-protein
,	NN	O	O
called	NN	O	O
inhibitor	NN	O	B-protein
of	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
from	NN	O	O
a	NN	O	O
sequestered	NN	O	O
cytoplasmic	NN	O	O
form	NN	O	O
.	NN	O	O

This	NN	O	O
event	NN	O	O
is	NN	O	O
also	NN	O	O
triggered	NN	O	O
under	NN	O	O
various	NN	O	O
other	NN	O	O
conditions	NN	O	O
of	NN	O	O
biomedical	NN	O	O
importance	NN	O	O
.	NN	O	O

Other	NN	O	O
bacterial	NN	O	O
toxins	NN	O	O
,	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
interleukin-1	NN	O	B-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
mitogens	NN	O	I-protein
,	NN	O	O
UV	NN	O	O
light	NN	O	O
,	NN	O	O
gamma	NN	O	O
rays	NN	O	O
and	NN	O	O
oxidative	NN	O	O
stress	NN	O	O
were	NN	O	O
reported	NN	O	O
to	NN	O	O
induce	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
activated	NN	O	O
form	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
rapidly	NN	O	O
taken	NN	O	O
up	NN	O	O
into	NN	O	O
nuclei	NN	O	O
,	NN	O	O
initiates	NN	O	O
transcription	NN	O	O
from	NN	O	O
immediate	NN	O	O
early	NN	O	O
genes	NN	O	O
in	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Most	NN	O	O
of	NN	O	O
the	NN	O	O
target	NN	O	O
genes	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
are	NN	O	O
of	NN	O	O
relevance	NN	O	O
for	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
and	NN	O	O
can	NN	O	O
be	NN	O	O
grouped	NN	O	O
into	NN	O	O
those	NN	O	O
encoding	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
acute	NN	O	B-protein
phase	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
viral	NN	O	B-DNA
genomes	NN	O	I-DNA
,	NN	O	O
such	NN	O	O
as	NN	O	O
that	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
will	NN	O	O
discuss	NN	O	O
recent	NN	O	O
experimental	NN	O	O
evidences	NN	O	O
suggesting	NN	O	O
that	NN	O	O
LPS	NN	O	O
might	NN	O	O
share	NN	O	O
a	NN	O	O
pathway	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
with	NN	O	O
other	NN	O	O
inducers	NN	O	O
of	NN	O	O
the	NN	O	O
factor	NN	O	O
.	NN	O	O

This	NN	O	O
common	NN	O	O
pathway	NN	O	O
may	NN	O	O
involve	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
(	NN	O	O
ROI	NN	O	O
)	NN	O	O
as	NN	O	O
messenger	NN	O	O
molecules	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
The	NN	O	O
trend	NN	O	O
of	NN	O	O
molecular	NN	O	O
biology	NN	O	O
study	NN	O	O
on	NN	O	O
eosinophils	NN	O	B-cell_type
]	NN	O	O

Recently	NN	O	O
,	NN	O	O
many	NN	O	O
investigators	NN	O	O
have	NN	O	O
been	NN	O	O
interested	NN	O	O
in	NN	O	O
the	NN	O	O
study	NN	O	O
on	NN	O	O
eosinophil	NN	O	B-cell_type
biology	NN	O	O
since	NN	O	O
genes	NN	O	O
association	NN	O	O
with	NN	O	O
eosinophils	NN	O	B-cell_type
such	NN	O	O
as	NN	O	O
interleukin-5	NN	O	B-protein
or	NN	O	O
eosinophil	NN	O	B-protein
granule	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
EPO	NN	O	B-protein
,	NN	O	O
ECP	NN	O	B-protein
,	NN	O	O
EDN	NN	O	B-protein
,	NN	O	O
MBP	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CLC	NN	O	B-protein
)	NN	O	O
,	NN	O	O
were	NN	O	O
isolated	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
commitment	NN	O	O
of	NN	O	O
progenitors	NN	O	O
to	NN	O	O
the	NN	O	O
eosinophil	NN	O	B-cell_type
lineage	NN	O	I-cell_type
has	NN	O	O
not	NN	O	O
been	NN	O	O
determined	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
eosinophil-specific	NN	O	B-DNA
genes	NN	O	I-DNA
encoding	NN	O	O
primary	NN	O	O
and	NN	O	O
secondary	NN	O	B-protein
granule	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
e.g	NN	O	O
.	NN	O	O
ECP	NN	O	B-protein
,	NN	O	O
EDN	NN	O	B-protein
,	NN	O	O
EPO	NN	O	B-protein
,	NN	O	O
MBP	NN	O	B-protein
,	NN	O	O
and	NN	O	O
CLC	NN	O	B-protein
)	NN	O	O
are	NN	O	O
expressed	NN	O	O
and	NN	O	O
regulated	NN	O	O
during	NN	O	O
eosinophilopoiesis	NN	O	O
is	NN	O	O
also	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
,	NN	O	O
I	NN	O	O
described	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
genes	NN	O	O
encoding	NN	O	O
eosinophil	NN	O	B-protein
granule	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
the	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
binding	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
during	NN	O	O
eosinophil	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	O
and	NN	O	O
cytoplasmic	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
lymphoid-specific	NN	O	B-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
complex	NN	O	O
.	NN	O	O

The	NN	O	O
lymphoid-specific	NN	O	B-protein
transcription	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
early	NN	O	O
gene	NN	O	O
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
assembled	NN	O	O
from	NN	O	O
a	NN	O	O
pre-existing	NN	O	O
,	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
restricted	NN	O	I-protein
cytoplasmic	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
an	NN	O	O
inducible	NN	O	O
ubiquitous	NN	O	B-protein
nuclear	NN	O	B-protein
component	NN	O	I-protein
within	NN	O	O
30	NN	O	O
min	NN	O	O
after	NN	O	O
activation	NN	O	O
through	NN	O	O
the	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
implicated	NN	O	O
the	NN	O	O
family	NN	O	O
of	NN	O	O
AP1	NN	O	B-protein
factors	NN	O	I-protein
as	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
NF-AT	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Evidence	NN	O	O
is	NN	O	O
provided	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
nuclear	NN	O	O
component	NN	O	O
of	NN	O	O
human	NN	O	B-protein
NF-AT	NN	O	I-protein
contains	NN	O	O
the	NN	O	O
phorbol	NN	O	B-protein
ester-inducible	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
AP1	NN	O	I-protein
(	NN	O	O
Jun/Fos	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
characterize	NN	O	O
which	NN	O	O
AP1	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
can	NN	O	O
assume	NN	O	O
this	NN	O	O
role	NN	O	O
.	NN	O	O

Antisera	NN	O	O
to	NN	O	O
Fos	NN	O	B-protein
inhibits	NN	O	O
NF-AT	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
as	NN	O	O
does	NN	O	O
an	NN	O	O
oligonucleotide	NN	O	O
containing	NN	O	O
a	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	I-DNA
AP1	NN	O	I-DNA
.	NN	O	O

Constitutive	NN	O	O
expression	NN	O	O
in	NN	O	O
vivo	NN	O	O
of	NN	O	O
Fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
Fra-1	NN	O	B-protein
,	NN	O	O
eliminates	NN	O	O
the	NN	O	O
requirement	NN	O	O
for	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
stimulation	NN	O	O
,	NN	O	O
leaving	NN	O	O
NF-AT	NN	O	B-protein
-directed	NN	O	O
transcription	NN	O	O
responsive	NN	O	O
to	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
alone	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
cJun	NN	O	B-protein
or	NN	O	O
JunD	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
JunB	NN	O	B-protein
,	NN	O	O
also	NN	O	O
eliminates	NN	O	O
the	NN	O	O
requirement	NN	O	O
for	NN	O	O
PMA	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
many	NN	O	O
but	NN	O	O
not	NN	O	O
all	NN	O	O
Jun-	NN	O	B-protein
and	NN	O	I-protein
Fos-related	NN	O	I-protein
proteins	NN	O	I-protein
functionally	NN	O	O
activate	NN	O	O
NF-AT	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
component	NN	O	O
.	NN	O	O

NF-AT	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
can	NN	O	O
be	NN	O	O
reconstituted	NN	O	O
in	NN	O	O
vitro	NN	O	O
using	NN	O	O
semi-purified	NN	O	B-protein
AP1	NN	O	I-protein
proteins	NN	O	I-protein
mixed	NN	O	O
with	NN	O	O
cytosol	NN	O	O
from	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Fos	NN	O	O
proteins	NN	O	O
are	NN	O	O
not	NN	O	O
needed	NN	O	O
for	NN	O	O
this	NN	O	O
reconstitution	NN	O	O
,	NN	O	O
and	NN	O	O
although	NN	O	O
JunB	NN	O	B-protein
is	NN	O	O
not	NN	O	O
functional	NN	O	O
,	NN	O	O
it	NN	O	O
can	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
partially	NN	O	O
purified	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
component	NN	O	O
of	NN	O	O
NF-AT	NN	O	O
and	NN	O	O
show	NN	O	O
by	NN	O	O
elution	NN	O	O
and	NN	O	O
renaturation	NN	O	O
from	NN	O	O
SDS-polyacrylamide	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
gels	NN	O	O
that	NN	O	O
it	NN	O	O
has	NN	O	O
a	NN	O	O
molecular	NN	O	O
mass	NN	O	O
between	NN	O	O
94	NN	O	O
and	NN	O	O
116	NN	O	O
kDa	NN	O	O
and	NN	O	O
may	NN	O	O
have	NN	O	O
multiple	NN	O	O
differentially	NN	O	O
modified	NN	O	O
forms	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
sepsis	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
hormone-binding	NN	O	O
activity	NN	O	O
was	NN	O	O
studied	NN	O	O
by	NN	O	O
a	NN	O	O
whole-cell	NN	O	O
method	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
MNC	NN	O	B-cell_type
)	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
7	NN	O	O
patients	NN	O	O
during	NN	O	O
the	NN	O	O
hemodynamic	NN	O	O
compensatory	NN	O	O
phase	NN	O	O
of	NN	O	O
sepsis	NN	O	O
.	NN	O	O

4	NN	O	O
patients	NN	O	O
were	NN	O	O
receiving	NN	O	O
dopamine	NN	O	O
,	NN	O	O
which	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
count	NN	O	O
.	NN	O	O

The	NN	O	O
patients	NN	O	O
'	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
were	NN	O	O
normal	NN	O	O
or	NN	O	O
slightly	NN	O	O
elevated	NN	O	O
.	NN	O	O

Despite	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
,	NN	O	O
the	NN	O	O
mean	NN	O	O
GR	NN	O	B-protein
count	NN	O	O
and	NN	O	O
affinity	NN	O	O
in	NN	O	O
MNC	NN	O	B-cell_type
from	NN	O	O
septic	NN	O	O
patients	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
from	NN	O	O
those	NN	O	O
in	NN	O	O
normal	NN	O	O
controls	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
could	NN	O	O
still	NN	O	O
be	NN	O	O
effective	NN	O	O
in	NN	O	O
the	NN	O	O
hemodynamic	NN	O	O
compensatory	NN	O	O
phase	NN	O	O
of	NN	O	O
sepsis	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
impaired	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
subjects	NN	O	O
with	NN	O	O
some	NN	O	O
symptoms	NN	O	O
of	NN	O	O
premature	NN	O	O
aging	NN	O	O
.	NN	O	O

The	NN	O	O
study	NN	O	O
of	NN	O	O
human	NN	O	O
disorders	NN	O	O
known	NN	O	O
as	NN	O	O
premature	NN	O	O
aging	NN	O	O
syndromes	NN	O	O
may	NN	O	O
provide	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
cellular	NN	O	O
senescence	NN	O	O
.	NN	O	O

The	NN	O	O
main	NN	O	O
feature	NN	O	O
of	NN	O	O
cellular	NN	O	O
senescence	NN	O	O
in	NN	O	O
vitro	NN	O	O
is	NN	O	O
cessation	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Down	NN	O	O
syndrome	NN	O	O
(	NN	O	O
DS	NN	O	O
)	NN	O	O
and	NN	O	O
neuronal	NN	O	O
ceroid-lypofuscinosis	NN	O	O
(	NN	O	O
NCL	NN	O	O
)	NN	O	O
are	NN	O	O
clinically	NN	O	O
characterized	NN	O	O
by	NN	O	O
the	NN	O	O
premature	NN	O	O
onset	NN	O	O
of	NN	O	O
numerous	NN	O	O
features	NN	O	O
normally	NN	O	O
associated	NN	O	O
with	NN	O	O
human	NN	O	O
aging	NN	O	O
.	NN	O	O

Phytohemagglutinin	NN	O	B-protein
stimulated	NN	O	O
lymphocytes	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
DS	NN	O	O
subjects	NN	O	O
showed	NN	O	O
a	NN	O	O
statistically	NN	O	O
significant	NN	O	O
diminished	NN	O	O
proliferation	NN	O	O
capacity	NN	O	O
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
lymphocytes	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
NCL	NN	O	O
and	NN	O	O
healthy	NN	O	O
individuals	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
,	NN	O	O
by	NN	O	O
applying	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
slightly	NN	O	O
impaired	NN	O	O
AP-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
NCL	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
strong	NN	O	O
in	NN	O	O
DS	NN	O	B-cell_type
ones	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
same	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
proliferation	NN	O	O
cessation	NN	O	O
could	NN	O	O
exist	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
characterized	NN	O	O
by	NN	O	O
replicative	NN	O	O
senescence	NN	O	O
and	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
derived	NN	O	O
from	NN	O	O
individuals	NN	O	O
with	NN	O	O
premature	NN	O	O
aging	NN	O	O
syndromes	NN	O	O
(	NN	O	O
Down	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
contribution	NN	O	O
of	NN	O	O
herpes	NN	O	B-protein
simplex	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
gene	NN	O	I-protein
products	NN	O	I-protein
and	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
provirus	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
infection	NN	O	O
with	NN	O	O
herpes	NN	O	O
simplex	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HSV-1	NN	O	O
)	NN	O	O
activates	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
provirus	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
provirus	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
binding	NN	O	O
of	NN	O	O
55-	NN	O	B-protein
and	NN	O	I-protein
85-kDa	NN	O	I-protein
proteins	NN	O	I-protein
to	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
50-kDa	NN	O	B-protein
HLP-1	NN	O	I-protein
protein	NN	O	I-protein
to	NN	O	O
the	NN	O	O
LBP-1	NN	O	B-DNA
sequences	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

Further	NN	O	O
examination	NN	O	O
of	NN	O	O
this	NN	O	O
system	NN	O	O
has	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
HSV-1	NN	O	O
replication	NN	O	O
by	NN	O	O
the	NN	O	O
antiviral	NN	O	O
drug	NN	O	O
acyclovir	NN	O	O
does	NN	O	O
not	NN	O	O
inhibit	NN	O	O
HSV-1-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
provirus	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
HLP-1	NN	O	O
binding	NN	O	O
activities	NN	O	O
were	NN	O	O
substantially	NN	O	O
inhibited	NN	O	O
in	NN	O	O
acyclovir-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
the	NN	O	O
transient-transfection	NN	O	O
assay	NN	O	O
,	NN	O	O
ICP0	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
ICP4	NN	O	B-protein
,	NN	O	O
activated	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
ICP0	NN	O	B-protein
was	NN	O	O
greatly	NN	O	O
enhanced	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
provirus	NN	O	O
involves	NN	O	O
cooperation	NN	O	O
between	NN	O	O
the	NN	O	O
HSV-1-activated	NN	O	B-protein
cellular	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
virus-encoded	NN	O	B-protein
transactivator	NN	O	I-protein
,	NN	O	O
ICP0	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
IL-2	NN	O	B-protein
is	NN	O	O
associated	NN	O	O
with	NN	O	O
secretion	NN	O	O
of	NN	O	O
bioactive	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
protein	NN	O	I-protein
by	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
involves	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
(	NN	O	O
Mo	NN	O	B-cell_type
)	NN	O	O
constitutively	NN	O	O
display	NN	O	O
the	NN	O	O
beta-chain	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-protein
alpha-chain	NN	O	I-protein
is	NN	O	O
not	NN	O	O
constitutive	NN	O	O
but	NN	O	O
inducible	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
human	NN	O	B-protein
IL-2	NN	O	I-protein
to	NN	O	O
its	NN	O	O
binding	NN	O	O
site	NN	O	O
leads	NN	O	O
to	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
macrophage	NN	O	B-DNA
CSF	NN	O	I-DNA
(	NN	O	I-DNA
M-CSF	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
Mo	NN	O	B-cell_type
resulting	NN	O	O
in	NN	O	O
accumulation	NN	O	O
of	NN	O	O
M-CSF	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
subsequent	NN	O	O
release	NN	O	O
of	NN	O	O
bioactive	NN	O	O
M-CSF	NN	O	B-protein
protein	NN	O	I-protein
as	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
ELISA	NN	O	O
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
induced	NN	O	O
release	NN	O	O
of	NN	O	O
an	NN	O	O
activity-stimulating	NN	O	O
growth	NN	O	O
of	NN	O	O
monocyte-type	NN	O	B-cell_line
colonies	NN	O	I-cell_line
by	NN	O	O
a	NN	O	O
neutralizing	NN	O	O
anti-M-CSF	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

Transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
IL-2	NN	O	B-protein
is	NN	O	O
preceded	NN	O	O
by	NN	O	O
enhanced	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
to	NN	O	O
its	NN	O	O
recognition	NN	O	O
sequence	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	O
'	NN	O	O
regulatory	NN	O	O
enhancer	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
M-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	I-DNA
herpes	NN	O	I-DNA
thymidine	NN	O	I-DNA
kinase	NN	O	I-DNA
)	NN	O	I-DNA
construct	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
,	NN	O	O
it	NN	O	O
is	NN	O	O
shown	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
by	NN	O	O
an	NN	O	O
IL-2-induced	NN	O	B-protein
monocyte-derived	NN	O	I-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
confers	NN	O	O
reporter	NN	O	O
gene	NN	O	O
(	NN	O	O
human	NN	O	O
growth	NN	O	O
hormone	NN	O	O
)	NN	O	O
activity	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
induces	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
M-CSF	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
blood-derived	NN	O	I-cell_type
Mo	NN	O	I-cell_type
and	NN	O	O
provide	NN	O	O
evidence	NN	O	O
for	NN	O	O
involvement	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-1	NN	O	B-DNA
beta	NN	O	I-DNA
(	NN	O	I-DNA
IL-1	NN	O	I-DNA
beta	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
mycobacterial	NN	O	B-protein
components	NN	O	I-protein
and	NN	O	O
lipopolysaccharide	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
two	NN	O	O
nuclear	NN	O	B-DNA
factor-IL6	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
cytokines	NN	O	O
interleukin-1	NN	O	O
beta	NN	O	O
(	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
)	NN	O	O
and	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor	NN	O	O
alpha	NN	O	O
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
are	NN	O	O
released	NN	O	O
by	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
after	NN	O	O
stimulation	NN	O	O
with	NN	O	O
mycobacteria	NN	O	O
and	NN	O	O
are	NN	O	O
considered	NN	O	O
to	NN	O	O
mediate	NN	O	O
pathophysiologic	NN	O	O
events	NN	O	O
,	NN	O	O
including	NN	O	O
granuloma	NN	O	O
formation	NN	O	O
and	NN	O	O
systemic	NN	O	O
symptoms	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
Mycobacterium	NN	O	O
tuberculosis	NN	O	O
cell	NN	O	O
wall	NN	O	O
component	NN	O	O
lipoarabinomannan	NN	O	O
(	NN	O	O
LAM	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
very	NN	O	O
potent	NN	O	O
inducer	NN	O	O
of	NN	O	O
IL-1	NN	O	B-DNA
beta	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
investigated	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
this	NN	O	O
effect	NN	O	O
.	NN	O	O

We	NN	O	O
localized	NN	O	O
the	NN	O	O
LAM-	NN	O	O
,	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-	NN	O	O
,	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
-inducible	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
to	NN	O	O
a	NN	O	O
-131/+15	NN	O	O
(	NN	O	O
positions	NN	O	O
-131	NN	O	O
to	NN	O	O
+15	NN	O	O
)	NN	O	O
DNA	NN	O	O
fragment	NN	O	O
of	NN	O	O
the	NN	O	O
IL-1	NN	O	B-DNA
beta	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
deletion	NN	O	O
analysis	NN	O	O
and	NN	O	O
chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
assay	NN	O	O
.	NN	O	O

Within	NN	O	O
this	NN	O	O
DNA	NN	O	O
fragment	NN	O	O
,	NN	O	O
there	NN	O	O
were	NN	O	O
two	NN	O	O
novel	NN	O	O
9-bp	NN	O	O
motifs	NN	O	O
(	NN	O	O
-90/-82	NN	O	O
and	NN	O	O
-40/-32	NN	O	O
)	NN	O	O
with	NN	O	O
high	NN	O	O
homology	NN	O	O
to	NN	O	O
the	NN	O	O
nuclear	NN	O	B-DNA
factor-IL6	NN	O	I-DNA
(	NN	O	I-DNA
NF-IL6	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
NF-IL-6	NN	O	B-DNA
motifs	NN	O	I-DNA
could	NN	O	O
be	NN	O	O
independently	NN	O	O
activated	NN	O	O
by	NN	O	O
LAM	NN	O	O
,	NN	O	O
LPS	NN	O	O
,	NN	O	O
or	NN	O	O
TNF-alpha	NN	O	B-protein
and	NN	O	O
that	NN	O	O
they	NN	O	O
acted	NN	O	O
in	NN	O	O
an	NN	O	O
orientation-independent	NN	O	O
manner	NN	O	O
.	NN	O	O

DNA	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
revealed	NN	O	O
specific	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
from	NN	O	O
LAM-	NN	O	B-cell_line
,	NN	O	I-cell_line
LPS-	NN	O	I-cell_line
,	NN	O	I-cell_line
or	NN	O	I-cell_line
TNF-alpha-	NN	O	I-cell_line
stimulated	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
the	NN	O	O
NF-IL6	NN	O	O
motifs	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
NF-IL6	NN	O	B-DNA
sites	NN	O	I-DNA
mediate	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
stimuli	NN	O	O
LAM	NN	O	O
,	NN	O	O
LPS	NN	O	O
,	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

HIV-1	NN	O	B-protein
Nef	NN	O	I-protein
protein	NN	O	I-protein
inhibits	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
,	NN	O	O
HIV-1-LTR	NN	O	B-DNA
,	NN	O	O
contains	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
several	NN	O	O
cellular	NN	O	O
transcription	NN	O	O
factors	NN	O	O
which	NN	O	O
contribute	NN	O	O
to	NN	O	O
HIV-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
studies	NN	O	O
on	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1-encoded	NN	O	B-protein
Nef	NN	O	I-protein
protein	NN	O	I-protein
suggested	NN	O	O
that	NN	O	O
Nef	NN	O	B-protein
may	NN	O	O
be	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
Nef	NN	O	B-protein
affects	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
implicated	NN	O	O
in	NN	O	O
HIV-1	NN	O	O
regulation	NN	O	O
,	NN	O	O
32P-labeled	NN	O	O
oligonucleotides	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
binding	NN	O	O
sites	NN	O	O
were	NN	O	O
incubated	NN	O	O
with	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
from	NN	O	O
Nef-expressing	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
were	NN	O	O
not	NN	O	O
stimulated	NN	O	O
or	NN	O	O
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
T-cell	NN	O	B-protein
mitogens	NN	O	I-protein
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
Nef	NN	O	B-protein
inhibited	NN	O	O
the	NN	O	O
recruitment	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
mitogen-stimulated	NN	O	O
human	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
Nef	NN	O	B-cell_line
expressing	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
plasmid	NN	O	O
in	NN	O	O
which	NN	O	O
HIV-1	NN	O	B-DNA
AP-1	NN	O	I-DNA
DNA	NN	O	I-DNA
recognition	NN	O	I-DNA
sequences	NN	O	I-DNA
were	NN	O	O
cloned	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
(	NN	O	O
CAT	NN	O	B-protein
)	NN	O	O
gene	NN	O	O
.	NN	O	O

Mitogen-mediated	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
CAT	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
this	NN	O	O
construct	NN	O	O
was	NN	O	O
inhibited	NN	O	O
in	NN	O	O
Nef-expressing	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
control	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
,	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
AP-1	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
Nef	NN	O	B-protein
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
HIV-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
infected	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
locus	NN	O	I-DNA
.	NN	O	O

Transcription	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
gene	NN	O	I-DNA
cluster	NN	O	I-DNA
depends	NN	O	O
upon	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
sequences	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
are	NN	O	O
collectively	NN	O	O
termed	NN	O	O
the	NN	O	O
locus	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Recent	NN	O	O
studies	NN	O	O
have	NN	O	O
provided	NN	O	O
new	NN	O	O
insights	NN	O	O
into	NN	O	O
how	NN	O	O
the	NN	O	O
individual	NN	O	O
genes	NN	O	O
of	NN	O	O
the	NN	O	O
cluster	NN	O	O
are	NN	O	O
regulated	NN	O	O
through	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
crux	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
is	NN	O	O
how	NN	O	O
the	NN	O	O
locus	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
communicates	NN	O	O
with	NN	O	O
the	NN	O	O
gene-proximal	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Enhancing	NN	O	O
effect	NN	O	O
of	NN	O	O
17	NN	O	O
beta-estradiol	NN	O	O
on	NN	O	O
human	NN	O	O
NK	NN	O	O
cell	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
in	NN	O	O
vitro	NN	O	O
effect	NN	O	O
of	NN	O	O
17	NN	O	O
beta-estradiol	NN	O	O
on	NN	O	O
NK	NN	O	O
activity	NN	O	O
was	NN	O	O
studied	NN	O	O
.	NN	O	O

The	NN	O	O
proliferation	NN	O	O
and	NN	O	O
NK	NN	O	O
activity	NN	O	O
of	NN	O	O
YT-N17	NN	O	B-cell_line
(	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
NK-like	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
were	NN	O	O
enhanced	NN	O	O
by	NN	O	O
17	NN	O	O
beta-estradiol	NN	O	O
(	NN	O	O
E2	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
enhancement	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
tamoxifen	NN	O	O
(	NN	O	O
Tx	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
antagonist	NN	O	O
of	NN	O	O
E2	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
contrary	NN	O	O
,	NN	O	O
other	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
such	NN	O	O
as	NN	O	O
Tx	NN	O	O
,	NN	O	O
progesterone	NN	O	O
,	NN	O	O
and	NN	O	O
testosterone	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

YT-N17	NN	O	B-cell_line
contained	NN	O	O
11.8	NN	O	O
fmol/mg	NN	O	O
protein	NN	O	O
of	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
mean	NN	O	O
of	NN	O	O
two	NN	O	O
independent	NN	O	O
assays	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
value	NN	O	O
which	NN	O	O
was	NN	O	O
5-10-fold	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
other	NN	O	O
hematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

An	NN	O	O
enhancement	NN	O	O
of	NN	O	O
NK	NN	O	O
activity	NN	O	O
by	NN	O	O
E2	NN	O	O
was	NN	O	O
also	NN	O	O
seen	NN	O	O
in	NN	O	O
large	NN	O	B-cell_type
granular	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
normal	NN	O	O
subjects	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
was	NN	O	O
again	NN	O	O
suppressed	NN	O	O
by	NN	O	O
Tx	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
E2	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
activating	NN	O	O
factors	NN	O	O
for	NN	O	O
NK/LGL	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
serum	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	I-DNA
NF-Y	NN	O	I-DNA
mediate	NN	O	O
the	NN	O	O
serum	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
thrombospondin	NN	O	I-DNA
1	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
thrombospondin	NN	O	B-protein
1	NN	O	I-protein
(	NN	O	O
TSP	NN	O	B-protein
1	NN	O	I-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
TSP	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
by	NN	O	O
growth	NN	O	O
factors	NN	O	O
.	NN	O	O

We	NN	O	O
tested	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
TSP	NN	O	I-DNA
1-chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
constructs	NN	O	I-DNA
to	NN	O	O
respond	NN	O	O
to	NN	O	O
serum	NN	O	O
in	NN	O	O
stably	NN	O	O
transfected	NN	O	O
NIH-3T3	NN	O	O
cells	NN	O	O
.	NN	O	O

Two	NN	O	O
transcriptional	NN	O	O
elements	NN	O	O
in	NN	O	O
the	NN	O	O
TSP	NN	O	B-protein
1	NN	O	I-protein
promoter	NN	O	O
,	NN	O	O
a	NN	O	O
distal	NN	O	O
element	NN	O	O
at	NN	O	O
-1280	NN	O	O
and	NN	O	O
a	NN	O	O
proximal	NN	O	O
element	NN	O	O
at	NN	O	O
-65	NN	O	O
,	NN	O	O
were	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
TSP	NN	O	O
1	NN	O	O
gene	NN	O	O
to	NN	O	O
serum	NN	O	O
.	NN	O	O

The	NN	O	O
distal	NN	O	O
element	NN	O	O
contains	NN	O	O
the	NN	O	O
5'-CC	NN	O	O
(	NN	O	O
A	NN	O	O
+	NN	O	O
T	NN	O	O
)	NN	O	O
6GG-3	NN	O	O
'	NN	O	O
consensus	NN	O	O
sequence	NN	O	O
characteristic	NN	O	O
of	NN	O	O
a	NN	O	O
serum-response	NN	O	O
element	NN	O	O
(	NN	O	O
SRE	NN	O	O
)	NN	O	O
.	NN	O	O

Deletions	NN	O	O
or	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
element	NN	O	O
reduced	NN	O	O
the	NN	O	O
serum	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
TSP	NN	O	B-DNA
1	NN	O	I-DNA
gene	NN	O	I-DNA
by	NN	O	O
80-90	NN	O	O
%	NN	O	O
.	NN	O	O

In	NN	O	O
gel-shift	NN	O	O
assays	NN	O	O
,	NN	O	O
the	NN	O	O
-1280	NN	O	B-DNA
element	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
SRE	NN	O	I-DNA
cross	NN	O	I-DNA
-competed	NN	O	O
,	NN	O	O
whereas	NN	O	O
their	NN	O	O
functional	NN	O	O
and	NN	O	O
binding	NN	O	B-DNA
mutants	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
.	NN	O	O

The	NN	O	O
proximal	NN	O	B-DNA
element	NN	O	I-DNA
contains	NN	O	O
the	NN	O	O
sequence	NN	O	O
5'-GGCCAATGGG-3	NN	O	O
'	NN	O	O
,	NN	O	O
which	NN	O	O
closely	NN	O	O
resembles	NN	O	O
the	NN	O	O
consensus	NN	O	O
binding	NN	O	O
motif	NN	O	O
for	NN	O	O
the	NN	O	O
CCAAT-binding	NN	O	B-DNA
factor	NN	O	I-DNA
NF-Y	NN	O	B-protein
(	NN	O	O
CBF	NN	O	B-protein
,	NN	O	O
CP1	NN	O	B-protein
,	NN	O	O
alpha	NN	O	B-protein
CP1	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Deletions	NN	O	O
or	NN	O	O
mutations	NN	O	O
in	NN	O	O
this	NN	O	O
element	NN	O	O
also	NN	O	O
reduced	NN	O	O
the	NN	O	O
serum	NN	O	O
response	NN	O	O
by	NN	O	O
80-90	NN	O	O
%	NN	O	O
.	NN	O	O

Methylation	NN	O	O
interference	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
-65	NN	O	B-DNA
region	NN	O	I-DNA
identified	NN	O	O
a	NN	O	O
pattern	NN	O	O
of	NN	O	O
contacts	NN	O	O
with	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
resembling	NN	O	O
that	NN	O	O
for	NN	O	O
NF-Y	NN	O	B-protein
,	NN	O	O
and	NN	O	O
an	NN	O	O
NF-Y-binding	NN	O	B-DNA
site	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
TSP	NN	O	I-DNA
1	NN	O	I-DNA
element	NN	O	I-DNA
cross-competed	NN	O	O
in	NN	O	O
gel-shift	NN	O	O
assays	NN	O	O
,	NN	O	O
whereas	NN	O	O
their	NN	O	O
binding	NN	O	B-DNA
mutants	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
an	NN	O	O
abbreviated	NN	O	B-DNA
TSP	NN	O	I-DNA
1	NN	O	I-DNA
promoter/5'-flank	NN	O	I-DNA
,	NN	O	O
containing	NN	O	O
the	NN	O	O
SRE-	NN	O	B-DNA
and	NN	O	I-DNA
NF-Y-binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
mediated	NN	O	O
a	NN	O	O
serum	NN	O	O
response	NN	O	O
that	NN	O	O
was	NN	O	O
close	NN	O	O
in	NN	O	O
magnitude	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
parent	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
serum	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
TSP	NN	O	B-DNA
1	NN	O	I-DNA
gene	NN	O	I-DNA
requires	NN	O	O
the	NN	O	O
coordinated	NN	O	O
function	NN	O	O
of	NN	O	O
an	NN	O	O
SRE-	NN	O	B-DNA
and	NN	O	I-DNA
NF-Y-binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Cell-type-specific	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
parathyroid	NN	O	B-DNA
hormone-related	NN	O	I-DNA
protein	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
I	NN	O	I-protein
(	NN	O	I-protein
HTLV-I	NN	O	I-protein
)	NN	O	I-protein
tax	NN	O	I-protein
and	NN	O	O
HTLV-II	NN	O	B-protein
tax	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
leukemia	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
I	NN	O	I-protein
(	NN	O	I-protein
HTLV-I	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	I-protein
HTLV-II	NN	O	I-protein
Tax	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
potent	NN	O	O
transactivators	NN	O	B-protein
of	NN	O	O
viral	NN	O	O
and	NN	O	O
cellular	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Using	NN	O	O
deletion	NN	O	B-DNA
mutants	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
downstream	NN	O	O
parathyroid	NN	O	B-DNA
hormone-related	NN	O	I-DNA
protein	NN	O	I-DNA
(	NN	O	I-DNA
PTHrP	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
responsive	NN	O	O
to	NN	O	O
both	NN	O	O
HTLV-I	NN	O	B-protein
and	NN	O	I-protein
HTLV-II	NN	O	I-protein
Tax	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
AP1/c-jun	NN	O	B-DNA
proto-oncogene	NN	O	I-DNA
.	NN	O	O

Transactivation	NN	O	O
of	NN	O	O
PTHrP	NN	O	B-protein
by	NN	O	O
Tax	NN	O	B-protein
was	NN	O	O
seen	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
or	NN	O	O
fibroblasts	NN	O	B-cell_line
.	NN	O	O

A	NN	O	O
carboxy	NN	O	B-DNA
terminal	NN	O	I-DNA
Tax	NN	O	I-DNA
deletion	NN	O	I-DNA
mutant	NN	O	I-DNA
was	NN	O	O
deficient	NN	O	O
in	NN	O	O
transactivation	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
PTHrP	NN	O	B-DNA
and	NN	O	I-DNA
IL2R	NN	O	I-DNA
alpha	NN	O	I-DNA
promoters	NN	O	I-DNA
but	NN	O	O
not	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Exogenous	NN	O	O
provision	NN	O	O
of	NN	O	O
NFkB	NN	O	B-protein
rescued	NN	O	O
IL2R	NN	O	B-protein
alpha	NN	O	O
expression	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
PTHrP	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
HTLV-I	NN	O	B-protein
Tax	NN	O	I-protein
,	NN	O	O
HTLV-II	NN	O	B-protein
Tax	NN	O	I-protein
,	NN	O	O
and	NN	O	O
c-jun	NN	O	B-protein
transactivate	NN	O	O
PTHrP	NN	O	B-protein
and	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
hypercalcemia	NN	O	O
in	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
Alu	NN	O	B-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
cell-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
gamma	NN	O	B-protein
chain	NN	O	I-protein
of	NN	O	O
Fc	NN	O	B-protein
and	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Fc	NN	O	B-protein
epsilon	NN	O	I-protein
RI-gamma	NN	O	I-protein
chains	NN	O	I-protein
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
where	NN	O	O
they	NN	O	O
play	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

They	NN	O	O
are	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
IgE	NN	O	I-protein
receptor	NN	O	I-protein
in	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
basophils	NN	O	B-cell_type
,	NN	O	O
Langerhans	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
possibly	NN	O	O
other	NN	O	O
cells	NN	O	O
;	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
low	NN	O	O
affinity	NN	O	O
receptor	NN	O	O
for	NN	O	O
IgG	NN	O	B-protein
(	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RIIIA	NN	O	I-protein
or	NN	O	O
CD16	NN	O	B-protein
)	NN	O	O
in	NN	O	O
natural	NN	O	O
killer	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
;	NN	O	O
and	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
in	NN	O	O
subsets	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
gamma	NN	O	B-protein
chain	NN	O	I-protein
gene	NN	O	O
by	NN	O	O
analyzing	NN	O	O
the	NN	O	O
2.5-kilobase	NN	O	B-DNA
sequence	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
start	NN	O	O
site	NN	O	O
.	NN	O	O

This	NN	O	O
sequence	NN	O	O
contains	NN	O	O
a	NN	O	O
promoter	NN	O	B-DNA
specific	NN	O	O
to	NN	O	O
cells	NN	O	B-cell_type
of	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
tissue	NN	O	O
specificity	NN	O	O
of	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
is	NN	O	O
only	NN	O	O
partial	NN	O	O
because	NN	O	O
it	NN	O	O
is	NN	O	O
active	NN	O	O
in	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
tested	NN	O	O
here	NN	O	O
,	NN	O	O
regardless	NN	O	O
of	NN	O	O
whether	NN	O	O
they	NN	O	O
constitutively	NN	O	O
express	NN	O	O
Fc	NN	O	O
epsilon	NN	O	O
RI-	NN	O	O
gamma	NN	O	B-protein
chain	NN	O	I-protein
transcripts	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
two	NN	O	O
adjacent	NN	O	O
cis-acting	NN	O	O
regulatory	NN	O	O
elements	NN	O	O
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
part	NN	O	O
of	NN	O	O
an	NN	O	O
Alu	NN	O	O
repeat	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
(	NN	O	O
-445/-366	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
positive	NN	O	O
element	NN	O	O
active	NN	O	O
in	NN	O	O
both	NN	O	O
basophils	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
second	NN	O	O
(	NN	O	O
-365/-264	NN	O	O
)	NN	O	O
binds	NN	O	O
to	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
which	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
different	NN	O	O
in	NN	O	O
basophils	NN	O	B-cell_type
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
negative	NN	O	O
element	NN	O	O
in	NN	O	O
basophils	NN	O	B-cell_type
and	NN	O	O
as	NN	O	O
a	NN	O	O
positive	NN	O	O
one	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
this	NN	O	O
Alu	NN	O	B-DNA
repeat	NN	O	I-DNA
(	NN	O	O
90	NN	O	O
%	NN	O	O
identical	NN	O	O
to	NN	O	O
Alu	NN	O	B-DNA
consensus	NN	O	I-DNA
sequences	NN	O	I-DNA
)	NN	O	O
has	NN	O	O
evolved	NN	O	O
to	NN	O	O
become	NN	O	O
both	NN	O	O
a	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	O
regulator	NN	O	O
.	NN	O	O

-DOCSTART-	O

Chlorinated	NN	O	O
dibenzo-p-dioxins	NN	O	O
and	NN	O	O
dibenzofurans	NN	O	O
and	NN	O	O
the	NN	O	O
human	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

1	NN	O	O
.	NN	O	O
Blood	NN	O	B-protein
cell	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
volunteers	NN	O	O
with	NN	O	O
moderately	NN	O	O
increased	NN	O	O
body	NN	O	O
burdens	NN	O	O
.	NN	O	O

Using	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
(	NN	O	O
mAbs	NN	O	B-protein
)	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
surface	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
lymphocyte	NN	O	B-cell_type
subpopulations	NN	O	I-cell_type
of	NN	O	O
workers	NN	O	O
with	NN	O	O
moderately	NN	O	O
increased	NN	O	O
body	NN	O	O
burdens	NN	O	O
of	NN	O	O
2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8-tetrachlorodibenzo-p-dioxin	NN	O	O
(	NN	O	O
TCDD	NN	O	O
)	NN	O	O
and	NN	O	O
of	NN	O	O
other	NN	O	O
polychlorinated	NN	O	O
dibenzo-p-dioxins	NN	O	O
and	NN	O	O
dibenzofurans	NN	O	O
(	NN	O	O
PCDD/PCDF	NN	O	O
)	NN	O	O
,	NN	O	O
expressed	NN	O	O
here	NN	O	O
as	NN	O	O
International-Toxicity	NN	O	O
Equivalencies	NN	O	O
(	NN	O	O
I-TE	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
hypothesis	NN	O	O
to	NN	O	O
be	NN	O	O
tested	NN	O	O
was	NN	O	O
whether	NN	O	O
or	NN	O	O
not	NN	O	O
humans	NN	O	O
exhibit	NN	O	O
a	NN	O	O
similar	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
PCDDs/PCDFs	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
surface	NN	O	B-protein
receptors	NN	O	I-protein
found	NN	O	O
previously	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
small	NN	O	O
doses	NN	O	O
of	NN	O	O
2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
7	NN	O	O
,	NN	O	O
8-tetrachlorodibenzo-p-dioxin	NN	O	O
(	NN	O	O
TCDD	NN	O	O
)	NN	O	O
in	NN	O	O
Callithrix	NN	O	O
jacchus	NN	O	O
.	NN	O	O

These	NN	O	O
are	NN	O	O
:	NN	O	O
helper-inducer	NN	O	B-cell_type
(	NN	O	I-cell_type
memory	NN	O	I-cell_type
)	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
CD4+CD45R0+CD45RA-CD29highCD11a+	NN	O	B-cell_type
)	NN	O	O
,	NN	O	O
CD20+	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
cytotoxic	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
CD8+CD56+/CD57+	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
68	NN	O	O
triple-labellings	NN	O	O
with	NN	O	O
mAbs	NN	O	B-protein
were	NN	O	O
performed	NN	O	O
on	NN	O	O
the	NN	O	O
cells	NN	O	O
of	NN	O	O
each	NN	O	O
volunteer	NN	O	O
to	NN	O	O
possibly	NN	O	O
generate	NN	O	O
further	NN	O	O
hypotheses	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
evaluated	NN	O	O
whether	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
variables	NN	O	O
might	NN	O	O
be	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
biomarker	NN	O	O
of	NN	O	O
effects	NN	O	O
for	NN	O	O
this	NN	O	O
class	NN	O	O
of	NN	O	O
compounds	NN	O	O
.	NN	O	O

There	NN	O	O
were	NN	O	O
two	NN	O	O
main	NN	O	O
goals	NN	O	O
:	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
to	NN	O	O
evaluate	NN	O	O
whether	NN	O	O
workers	NN	O	O
with	NN	O	O
a	NN	O	O
moderately	NN	O	O
increased	NN	O	O
PCDD/PCDF-body	NN	O	O
burden	NN	O	O
[	NN	O	O
25-140	NN	O	O
ppt	NN	O	O
TCDD	NN	O	O
or	NN	O	O
104-522	NN	O	O
ppt	NN	O	O
I-TE	NN	O	O
in	NN	O	O
blood	NN	O	O
fat	NN	O	O
]	NN	O	O
exhibit	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
surface	NN	O	B-protein
receptors	NN	O	I-protein
of	NN	O	O
white	NN	O	B-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
observed	NN	O	O
in	NN	O	O
previous	NN	O	O
studies	NN	O	O
in	NN	O	O
non-human	NN	O	O
primates	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
to	NN	O	O
clarify	NN	O	O
whether	NN	O	O
persons	NN	O	O
at	NN	O	O
the	NN	O	O
upper	NN	O	O
range	NN	O	O
[	NN	O	O
10-23	NN	O	O
ppt	NN	O	O
TCDD	NN	O	O
or	NN	O	O
30-90	NN	O	O
ppt	NN	O	O
I-TE	NN	O	O
in	NN	O	O
blood	NN	O	O
fat	NN	O	O
]	NN	O	O
of	NN	O	O
the	NN	O	O
body	NN	O	O
burden	NN	O	O
reference	NN	O	O
values	NN	O	O
of	NN	O	O
a	NN	O	O
not	NN	O	O
particularly	NN	O	O
exposed	NN	O	O
population	NN	O	O
show	NN	O	O
detectable	NN	O	O
deviations	NN	O	O
in	NN	O	O
these	NN	O	O
immunological	NN	O	O
variables	NN	O	O
,	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
persons	NN	O	O
at	NN	O	O
the	NN	O	O
lower	NN	O	O
and	NN	O	O
medium	NN	O	O
range	NN	O	O
[	NN	O	O
1-3	NN	O	O
ppt	NN	O	O
TCDD	NN	O	O
or	NN	O	O
9-29	NN	O	O
ppt	NN	O	O
I-TE	NN	O	O
]	NN	O	O
of	NN	O	O
these	NN	O	O
body	NN	O	O
burden	NN	O	O
reference	NN	O	O
values	NN	O	O
.	NN	O	O

Regression	NN	O	O
analysis	NN	O	O
of	NN	O	O
our	NN	O	O
data	NN	O	O
revealed	NN	O	O
slight	NN	O	O
trends	NN	O	O
for	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
biomarkers	NN	O	O
(	NN	O	O
e.g.	NN	O	O
CD45R0+	NN	O	O
)	NN	O	O
.	NN	O	O

With	NN	O	O
one	NN	O	O
exception	NN	O	O
,	NN	O	O
these	NN	O	O
were	NN	O	O
all	NN	O	O
increases	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
the	NN	O	O
alterations	NN	O	O
observed	NN	O	O
are	NN	O	O
of	NN	O	O
medical	NN	O	O
relevance	NN	O	O
.	NN	O	O

The	NN	O	O
slight	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
CD4+CD45R0+	NN	O	B-cell_type
cells	NN	O	I-cell_type
remained	NN	O	O
significant	NN	O	O
even	NN	O	O
after	NN	O	O
covariant	NN	O	O
analysis	NN	O	O
taking	NN	O	O
age-related	NN	O	O
changes	NN	O	O
into	NN	O	O
account	NN	O	O
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
the	NN	O	O
data	NN	O	O
do	NN	O	O
not	NN	O	O
provide	NN	O	O
any	NN	O	O
evidence	NN	O	O
to	NN	O	O
support	NN	O	O
an	NN	O	O
assumption	NN	O	O
that	NN	O	O
moderately	NN	O	O
increased	NN	O	O
body	NN	O	O
burdens	NN	O	O
of	NN	O	O
PCDDs/PCDFs	NN	O	O
in	NN	O	O
adults	NN	O	O
induce	NN	O	O
decreases	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
components	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

Adult	NN	O	O
humans	NN	O	O
certainly	NN	O	O
are	NN	O	O
less	NN	O	O
susceptible	NN	O	O
to	NN	O	O
this	NN	O	O
action	NN	O	O
of	NN	O	O
PCDDs/PCDFs	NN	O	O
than	NN	O	O
adolescent	NN	O	O
Callithrix	NN	O	O
jacchus	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
latency	NN	O	O
reactivation	NN	O	O
by	NN	O	O
dehydroepiandrosterone	NN	O	O
(	NN	O	O
DHEA	NN	O	O
)	NN	O	O
and	NN	O	O
an	NN	O	O
analog	NN	O	O
of	NN	O	O
DHEA	NN	O	O
.	NN	O	O

The	NN	O	O
initial	NN	O	O
infection	NN	O	O
with	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
in	NN	O	O
most	NN	O	O
individuals	NN	O	O
usually	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
establishment	NN	O	O
of	NN	O	O
a	NN	O	O
latent	NN	O	O
or	NN	O	O
chronic	NN	O	O
infection	NN	O	O
before	NN	O	O
eventual	NN	O	O
progression	NN	O	O
toward	NN	O	O
acquired	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
.	NN	O	O

HIV-1	NN	O	O
can	NN	O	O
also	NN	O	O
establish	NN	O	O
a	NN	O	O
latent	NN	O	O
or	NN	O	O
persistent	NN	O	O
infection	NN	O	O
in	NN	O	O
some	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
show	NN	O	O
minimal	NN	O	O
constitutive	NN	O	O
virus	NN	O	O
expression	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
leading	NN	O	O
to	NN	O	O
enhanced	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
antigens	NN	O	B-protein
,	NN	O	O
mitogens	NN	O	O
,	NN	O	O
and	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
[	NN	O	O
TNF-alpha	NN	O	B-protein
]	NN	O	O
,	NN	O	O
interleukin	NN	O	B-protein
1	NN	O	I-protein
,	NN	O	O
and	NN	O	O
interleukin-2	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Various	NN	O	O
gene	NN	O	O
products	NN	O	O
from	NN	O	O
other	NN	O	O
viruses	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
,	NN	O	O
HSV	NN	O	O
,	NN	O	O
EBV	NN	O	O
,	NN	O	O
CMV	NN	O	O
,	NN	O	O
HBV	NN	O	O
,	NN	O	O
and	NN	O	O
HHV-6	NN	O	O
)	NN	O	O
can	NN	O	O
also	NN	O	O
enhance	NN	O	O
HIV-1	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
-driven	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
these	NN	O	O
observations	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
proposed	NN	O	O
that	NN	O	O
reactivation	NN	O	O
of	NN	O	O
latent	NN	O	O
HIV-1	NN	O	O
harbored	NN	O	O
in	NN	O	O
chronically	NN	O	B-cell_type
infected	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
or	NN	O	O
macrophages	NN	O	B-cell_type
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
AIDS	NN	O	O
.	NN	O	O

So	NN	O	O
far	NN	O	O
,	NN	O	O
there	NN	O	O
are	NN	O	O
no	NN	O	O
drugs	NN	O	O
or	NN	O	O
therapy	NN	O	O
available	NN	O	O
that	NN	O	O
can	NN	O	O
provide	NN	O	O
protection	NN	O	O
against	NN	O	O
HIV-1	NN	O	O
latency	NN	O	O
reactivation	NN	O	O
.	NN	O	O

ACH-2	NN	O	B-cell_line
,	NN	O	O
derived	NN	O	O
from	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
CEM	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
is	NN	O	O
chronically	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
with	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
constitutive	NN	O	O
virus	NN	O	O
expression	NN	O	O
.	NN	O	O

ACH-2	NN	O	B-cell_line
can	NN	O	O
be	NN	O	O
converted	NN	O	O
to	NN	O	O
productive	NN	O	O
infection	NN	O	O
by	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
,	NN	O	O
mitogen	NN	O	O
or	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
TNF-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
or	NN	O	O
infection	NN	O	O
with	NN	O	O
HSV	NN	O	O
.	NN	O	O

Therefore	NN	O	O
the	NN	O	O
ACH-2	NN	O	B-cell_line
cell	NN	O	O
line	NN	O	O
is	NN	O	O
a	NN	O	O
good	NN	O	O
candidate	NN	O	O
for	NN	O	O
studying	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
drugs	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
activation	NN	O	O
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
reported	NN	O	O
that	NN	O	O
DHEA	NN	O	O
and	NN	O	O
synthetic	NN	O	O
analogs	NN	O	O
of	NN	O	O
DHEA	NN	O	O
can	NN	O	O
be	NN	O	O
modest	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
IIIB	NN	O	O
replication	NN	O	O
in	NN	O	O
phytohemagglutinin-stimulated	NN	O	B-cell_line
peripheral	NN	O	I-cell_line
blood	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Occurrence	NN	O	O
of	NN	O	O
a	NN	O	O
silencer	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
naive	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
memory	NN	O	B-cell_type
resting	NN	O	I-cell_type
T	NN	O	I-cell_type
helper	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
the	NN	O	O
first	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
naive	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
by	NN	O	O
antigen	NN	O	O
is	NN	O	O
a	NN	O	O
key	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
shaping	NN	O	O
of	NN	O	O
the	NN	O	O
peripheral	NN	O	O
T	NN	O	O
cell	NN	O	O
specificity	NN	O	O
repertoire	NN	O	O
and	NN	O	O
maintenance	NN	O	O
of	NN	O	O
self-tolerance	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
activation	NN	O	O
shows	NN	O	O
that	NN	O	O
naive	NN	O	B-cell_type
human	NN	O	I-cell_type
helper	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
cord	NN	O	B-cell_type
blood	NN	O	I-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
adult	NN	O	B-cell_type
CD4+CD45RO-	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
regulate	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
by	NN	O	O
a	NN	O	O
mechanism	NN	O	O
involving	NN	O	O
both	NN	O	O
a	NN	O	O
silencer	NN	O	B-DNA
and	NN	O	O
an	NN	O	O
activator	NN	O	B-DNA
acting	NN	O	O
on	NN	O	O
the	NN	O	O
purine-rich	NN	O	B-DNA
IL-2	NN	O	I-DNA
promoter	NN	O	I-DNA
elements	NN	O	I-DNA
(	NN	O	O
NF-AT	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
memory	NN	O	O
cells	NN	O	O
,	NN	O	O
either	NN	O	O
in	NN	O	O
vitro	NN	O	B-cell_line
activated	NN	O	I-cell_line
helper	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
reverting	NN	O	O
to	NN	O	O
a	NN	O	O
resting	NN	O	O
state	NN	O	O
,	NN	O	O
or	NN	O	O
CD4+	NN	O	B-cell_line
T	NN	O	I-cell_line
(	NN	O	I-cell_line
memory	NN	O	I-cell_line
)	NN	O	I-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
or	NN	O	O
CD4+CD45RO+	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
isolated	NN	O	O
ex	NN	O	O
vivo	NN	O	O
,	NN	O	O
no	NN	O	O
longer	NN	O	O
have	NN	O	O
a	NN	O	O
silencer	NN	O	B-DNA
.	NN	O	O

Their	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
controlled	NN	O	O
solely	NN	O	O
by	NN	O	O
the	NN	O	O
transition	NN	O	O
from	NN	O	O
inactive	NN	O	O
to	NN	O	O
active	NN	O	O
functional	NN	O	O
state	NN	O	O
of	NN	O	O
a	NN	O	O
positive	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
these	NN	O	O
promoter	NN	O	O
elements	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
its	NN	O	O
cytoplasmic	NN	O	O
or	NN	O	O
nuclear	NN	O	O
location	NN	O	O
:	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
memory	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
the	NN	O	O
activator	NN	O	B-DNA
is	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
and	NN	O	O
is	NN	O	O
inactive	NN	O	O
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
stimulated	NN	O	B-cell_line
cells	NN	O	I-cell_line
it	NN	O	O
is	NN	O	O
functional	NN	O	O
in	NN	O	O
promoting	NN	O	O
transcription	NN	O	O
and	NN	O	O
now	NN	O	O
resides	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
the	NN	O	O
main	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
changes	NN	O	O
irreversibly	NN	O	O
after	NN	O	O
the	NN	O	O
first	NN	O	O
encounter	NN	O	O
of	NN	O	O
T	NN	O	O
cells	NN	O	O
with	NN	O	O
antigen	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
most	NN	O	O
likely	NN	O	O
that	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
silencer	NN	O	B-DNA
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
more	NN	O	O
stringent	NN	O	O
activation	NN	O	O
requirements	NN	O	O
of	NN	O	O
naive	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
the	NN	O	O
GATA-binding	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
eosinophils	NN	O	I-cell_type
and	NN	O	O
basophils	NN	O	B-cell_type
:	NN	O	O
potential	NN	O	O
role	NN	O	O
in	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
GATA-1	NN	O	B-protein
,	NN	O	O
GATA-2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GATA-3	NN	O	B-protein
was	NN	O	O
studied	NN	O	O
in	NN	O	O
eosinophils	NN	O	B-cell_type
and	NN	O	O
basophils	NN	O	B-cell_type
.	NN	O	O

Eosinophils	NN	O	B-cell_type
express	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
GATA-2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GATA-3	NN	O	B-protein
.	NN	O	O

Basophils	NN	O	B-cell_type
express	NN	O	O
GATA-2	NN	O	B-protein
and	NN	O	O
GATA-3	NN	O	B-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
eosinophilic	NN	O	I-cell_line
sublines	NN	O	I-cell_line
with	NN	O	O
either	NN	O	O
interleukin-5	NN	O	B-protein
or	NN	O	O
butyric	NN	O	O
acid	NN	O	O
increased	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
concomitant	NN	O	O
with	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
eosinophil-specific	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
whereas	NN	O	O
levels	NN	O	O
of	NN	O	O
GATA-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
remained	NN	O	O
relatively	NN	O	O
constant	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
for	NN	O	O
these	NN	O	O
proteins	NN	O	O
in	NN	O	O
eosinophils	NN	O	B-cell_type
and	NN	O	O
basophils	NN	O	B-cell_type
suggests	NN	O	O
that	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
these	NN	O	O
lineages	NN	O	O
may	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
GATA-binding	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Oxidoreductive	NN	O	O
regulation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Involvement	NN	O	O
of	NN	O	O
a	NN	O	O
cellular	NN	O	O
reducing	NN	O	O
catalyst	NN	O	O
thioredoxin	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
an	NN	O	O
oxidoreductive	NN	O	O
regulatory	NN	O	O
pathway	NN	O	O
for	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
pleiotropic	NN	O	B-protein
cellular	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
investigated	NN	O	O
by	NN	O	O
using	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
prepared	NN	O	O
from	NN	O	O
the	NN	O	O
nucleus	NN	O	O
and	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
the	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
cellular	NN	O	O
reducing	NN	O	O
catalyst	NN	O	O
thioredoxin	NN	O	B-protein
(	NN	O	O
Trx	NN	O	B-protein
)	NN	O	O
plays	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
stimulation	NN	O	O
of	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
-dependent	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
evidence	NN	O	O
suggesting	NN	O	O
that	NN	O	O
redox	NN	O	O
regulation	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
by	NN	O	O
Trx	NN	O	B-protein
might	NN	O	O
be	NN	O	O
exerted	NN	O	O
at	NN	O	O
a	NN	O	O
step	NN	O	O
after	NN	O	O
dissociation	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
molecule	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
cytosolic-anchoring	NN	O	B-protein
protein	NN	O	I-protein
for	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
Trx	NN	O	B-protein
in	NN	O	O
intact	NN	O	O
cells	NN	O	O
,	NN	O	O
we	NN	O	O
performed	NN	O	O
transient	NN	O	O
assay	NN	O	O
with	NN	O	O
a	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase-expressing	NN	O	I-DNA
plasmid	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
HIV	NN	O	I-DNA
)	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
and	NN	O	O
an	NN	O	O
effector	NN	O	B-DNA
plasmid	NN	O	I-DNA
expressing	NN	O	O
human	NN	O	B-protein
Trx	NN	O	I-protein
.	NN	O	O

The	NN	O	O
promoter	NN	O	O
activity	NN	O	O
from	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
was	NN	O	O
greatly	NN	O	O
augmented	NN	O	O
by	NN	O	O
co-transfecting	NN	O	O
the	NN	O	O
Trx-expressing	NN	O	B-DNA
plasmid	NN	O	I-DNA
,	NN	O	O
whose	NN	O	O
effect	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
-binding	NN	O	O
sites	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
cysteine	NN	O	O
residue	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
might	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
DNA-recognition	NN	O	O
by	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
that	NN	O	O
the	NN	O	O
redox	NN	O	O
control	NN	O	O
mechanism	NN	O	O
mediated	NN	O	O
by	NN	O	O
Trx	NN	O	B-protein
might	NN	O	O
have	NN	O	O
a	NN	O	O
regulatory	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
-mediated	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
may	NN	O	O
also	NN	O	O
provide	NN	O	O
a	NN	O	O
clue	NN	O	O
to	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
process	NN	O	O
of	NN	O	O
AIDS	NN	O	O
pathogenesis	NN	O	O
and	NN	O	O
its	NN	O	O
possible	NN	O	O
biochemical	NN	O	O
intervention	NN	O	O
.	NN	O	O

-DOCSTART-	O

Synergism	NN	O	O
between	NN	O	O
the	NN	O	O
CD3	NN	O	O
antigen-	NN	O	O
and	NN	O	O
CD2	NN	O	O
antigen-derived	NN	O	O
signals	NN	O	O
.	NN	O	O

Exploration	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
induction	NN	O	O
of	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
activity	NN	O	O
of	NN	O	O
cyclosporine	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
earlier	NN	O	O
that	NN	O	O
the	NN	O	O
crosslinkage	NN	O	O
of	NN	O	O
the	NN	O	O
CD3/TCR	NN	O	B-protein
complex	NN	O	I-protein
with	NN	O	O
the	NN	O	O
CD2	NN	O	B-protein
antigen	NN	O	I-protein
results	NN	O	O
in	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
this	NN	O	O
synergism	NN	O	O
was	NN	O	O
perceptible	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
process	NN	O	O
critical	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
understand	NN	O	O
the	NN	O	O
molecular	NN	O	O
and	NN	O	O
nuclear	NN	O	O
basis	NN	O	O
for	NN	O	O
this	NN	O	O
synergism	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
explored	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
highly	NN	O	O
purified	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
signaled	NN	O	O
via	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
and/or	NN	O	O
CD2	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
transmembrane	NN	O	O
signaling	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
ionomycin	NN	O	O
,	NN	O	O
and/or	NN	O	O
sn-1	NN	O	O
,	NN	O	O
2	NN	O	O
dioctanoyl	NN	O	O
glycerol	NN	O	O
,	NN	O	O
was	NN	O	O
also	NN	O	O
determined	NN	O	O
.	NN	O	O

The	NN	O	O
emergence	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
was	NN	O	O
investigated	NN	O	O
using	NN	O	O
interleukin-2	NN	O	B-DNA
sequence	NN	O	I-DNA
specific	NN	O	I-DNA
oligonucleotide	NN	O	I-DNA
probes	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
demonstrate	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
that	NN	O	O
CD3	NN	O	B-protein
antigen-derived	NN	O	O
signals	NN	O	O
and	NN	O	O
CD2	NN	O	B-protein
antigen	NN	O	I-protein
-derived	NN	O	O
signals	NN	O	O
are	NN	O	O
synergistic	NN	O	O
in	NN	O	O
inducing	NN	O	O
the	NN	O	O
emergence	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
NF-AT1	NN	O	B-DNA
,	NN	O	I-DNA
AP-1	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
NF-kB	NN	O	I-DNA
sites	NN	O	I-DNA
located	NN	O	O
in	NN	O	O
the	NN	O	O
promoter/enhancer	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
cyclosporine	NN	O	O
,	NN	O	O
at	NN	O	O
concentrations	NN	O	O
readily	NN	O	O
accomplished	NN	O	O
in	NN	O	O
clinical	NN	O	O
practice	NN	O	O
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
emergence	NN	O	O
of	NN	O	O
these	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
signaled	NN	O	O
via	NN	O	O
cell	NN	O	O
surface	NN	O	O
proteins	NN	O	O
implicated	NN	O	O
in	NN	O	O
antigen-dependent	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
by	NN	O	O
mobilization	NN	O	O
of	NN	O	O
cellular	NN	O	O
calcium	NN	O	O
and	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Induced	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
downregulation	NN	O	O
of	NN	O	O
erythroid	NN	O	B-protein
and	NN	O	I-protein
megakaryocytic	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
:	NN	O	O
a	NN	O	O
novel	NN	O	O
experimental	NN	O	O
model	NN	O	O
for	NN	O	O
hemopoietic	NN	O	O
lineage	NN	O	O
restriction	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-cell_line
erythroleukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
K562	NN	O	I-cell_line
can	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
along	NN	O	O
the	NN	O	O
erythroid	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
lineages	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
hexamethylene	NN	O	O
bisacetamide	NN	O	O
(	NN	O	O
HMBA	NN	O	O
)	NN	O	O
induced	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
differentiate	NN	O	O
along	NN	O	O
a	NN	O	O
third	NN	O	O
pathway	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
downregulation	NN	O	O
of	NN	O	O
two	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
normally	NN	O	O
expressed	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
,	NN	O	I-cell_type
mast	NN	O	I-cell_type
and	NN	O	I-cell_type
megakaryocyte	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

Northern	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
coordinate	NN	O	O
downregulation	NN	O	O
of	NN	O	O
alpha	NN	O	B-protein
globin	NN	O	I-protein
and	NN	O	O
gamma	NN	O	B-protein
globin	NN	O	I-protein
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
two	NN	O	O
lineage-restricted	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
SCL	NN	O	B-protein
and	NN	O	O
GATA-1	NN	O	B-protein
.	NN	O	O

Proliferation	NN	O	O
of	NN	O	O
the	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
also	NN	O	O
suppressed	NN	O	O
.	NN	O	O

Clonal	NN	O	O
assay	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
suppression	NN	O	O
was	NN	O	O
irreversible	NN	O	O
and	NN	O	O
appeared	NN	O	O
analogous	NN	O	O
to	NN	O	O
the	NN	O	O
commitment	NN	O	O
of	NN	O	O
murine	NN	O	B-cell_line
erythroleukemia	NN	O	I-cell_line
(	NN	O	I-cell_line
MEL	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
MEL	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
however	NN	O	O
,	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
acquired	NN	O	O
a	NN	O	O
macrophage-like	NN	O	O
morphology	NN	O	O
and	NN	O	O
exhibited	NN	O	O
a	NN	O	O
complete	NN	O	O
failure	NN	O	O
to	NN	O	O
generate	NN	O	O
benzidine-positive	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Electron	NN	O	O
microscopy	NN	O	O
revealed	NN	O	O
a	NN	O	O
marked	NN	O	O
increase	NN	O	O
in	NN	O	O
granules	NN	O	O
resembling	NN	O	O
those	NN	O	O
specific	NN	O	O
for	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

Surface	NN	O	O
marker	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
HMBA-induced	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressed	NN	O	O
reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
glycophorin	NN	O	B-protein
A	NN	O	I-protein
,	NN	O	O
CD5	NN	O	B-protein
,	NN	O	O
CD7	NN	O	B-protein
and	NN	O	O
CD11b	NN	O	B-protein
.	NN	O	O

No	NN	O	O
upregulation	NN	O	O
of	NN	O	O
megakaryocyte	NN	O	B-protein
or	NN	O	I-protein
lymphoid	NN	O	I-protein
markers	NN	O	I-protein
occurred	NN	O	O
.	NN	O	O

Thus	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
HMBA	NN	O	O
may	NN	O	O
provide	NN	O	O
a	NN	O	O
useful	NN	O	O
experimental	NN	O	O
system	NN	O	O
for	NN	O	O
studying	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
responsible	NN	O	O
for	NN	O	O
downmodulation	NN	O	O
of	NN	O	O
lineage-restricted	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
during	NN	O	O
hemopoietic	NN	O	O
lineage	NN	O	O
commitment	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
through	NN	O	O
CD2	NN	O	B-protein
plus	NN	O	I-protein
CD28	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
induces	NN	O	O
long-term	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
highly	NN	O	B-cell_type
purified	NN	O	I-cell_type
human	NN	O	I-cell_type
T-cells	NN	O	I-cell_type
via	NN	O	O
CD2	NN	O	B-protein
and	NN	O	I-protein
CD28	NN	O	I-protein
adhesion	NN	O	I-protein
molecules	NN	O	I-protein
induces	NN	O	O
and	NN	O	O
maintains	NN	O	O
proliferation	NN	O	O
for	NN	O	O
more	NN	O	O
than	NN	O	O
3	NN	O	O
weeks	NN	O	O
.	NN	O	O

This	NN	O	O
potent	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
-dependent	NN	O	O
activation	NN	O	O
does	NN	O	O
not	NN	O	O
require	NN	O	O
monocytes	NN	O	B-cell_type
or	NN	O	O
accessory	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Long-lasting	NN	O	O
IL-2	NN	O	B-protein
receptivity	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
high-level	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
inducible	NN	O	O
IL-2	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
chain	NN	O	I-DNA
(	NN	O	I-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
regulated	NN	O	O
at	NN	O	O
both	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
posttranscriptional	NN	O	O
levels	NN	O	O
.	NN	O	O

Increase	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
involves	NN	O	O
the	NN	O	O
enhanced	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
to	NN	O	O
its	NN	O	O
consensus	NN	O	B-DNA
sequence	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
5'-regulatory	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
dissect	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
unusually	NN	O	O
persistent	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
nuclear	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
a	NN	O	O
radiolabeled	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
kappa	NN	O	O
B-specific	NN	O	O
oligonucleotide	NN	O	O
probe	NN	O	O
during	NN	O	O
the	NN	O	O
time	NN	O	O
course	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
+	NN	O	O
CD28	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

Resting	NN	O	O
T-cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
contained	NN	O	O
KBF1/p50	NN	O	B-protein
homodimer	NN	O	I-protein
.	NN	O	O

After	NN	O	O
stimulation	NN	O	O
,	NN	O	O
two	NN	O	O
new	NN	O	O
kappa	NN	O	B-protein
B-specific	NN	O	I-protein
complexes	NN	O	I-protein
were	NN	O	O
identified	NN	O	O
as	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50-p65	NN	O	B-protein
heterodimer	NN	O	I-protein
and	NN	O	O
putative	NN	O	O
c-Rel	NN	O	B-protein
homodimer	NN	O	I-protein
or	NN	O	O
c-Rel-p65	NN	O	B-protein
heterodimer	NN	O	I-protein
.	NN	O	O

Both	NN	O	O
inducible	NN	O	O
complexes	NN	O	O
persisted	NN	O	O
for	NN	O	O
at	NN	O	O
least	NN	O	O
3	NN	O	O
weeks	NN	O	O
.	NN	O	O

Their	NN	O	O
relative	NN	O	O
levels	NN	O	O
were	NN	O	O
very	NN	O	O
similar	NN	O	O
for	NN	O	O
the	NN	O	O
duration	NN	O	O
of	NN	O	O
proliferation	NN	O	O
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
,	NN	O	O
CD2	NN	O	B-protein
+	NN	O	O
CD28	NN	O	B-protein
activation	NN	O	O
triggered	NN	O	O
a	NN	O	O
significant	NN	O	O
intracellular	NN	O	O
thiol	NN	O	O
decrease	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
oxygen	NN	O	O
radicals	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
micromolar	NN	O	O
amounts	NN	O	O
of	NN	O	O
pyrrolidine	NN	O	O
dithiocarbamate	NN	O	O
,	NN	O	O
an	NN	O	O
oxygen	NN	O	O
radical	NN	O	O
scavenger	NN	O	O
that	NN	O	O
efficiently	NN	O	O
blocked	NN	O	O
the	NN	O	O
nuclear	NN	O	O
appearance	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
T-lymphocytes	NN	O	B-cell_type
,	NN	O	O
also	NN	O	O
inhibited	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
,	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
long-term	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
primary	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
via	NN	O	O
CD2	NN	O	B-protein
+	NN	O	O
CD28	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Immobilization	NN	O	O
and	NN	O	O
recovery	NN	O	O
of	NN	O	O
fusion	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
B-lymphocyte	NN	O	B-cell_type
cells	NN	O	I-cell_type
using	NN	O	O
magnetic	NN	O	O
separation	NN	O	O
.	NN	O	O

A	NN	O	O
new	NN	O	O
approach	NN	O	O
to	NN	O	O
facilitate	NN	O	O
immobilization	NN	O	O
and	NN	O	O
affinity	NN	O	O
purification	NN	O	O
of	NN	O	O
recombinant	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
selected	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
developed	NN	O	O
.	NN	O	O

Using	NN	O	O
magnetic	NN	O	O
beads	NN	O	O
with	NN	O	O
attached	NN	O	O
DNA	NN	O	O
containing	NN	O	O
the	NN	O	O
Escherichia	NN	O	B-DNA
coli	NN	O	I-DNA
lac	NN	O	I-DNA
operator	NN	O	I-DNA
,	NN	O	O
fusion	NN	O	B-protein
proteins	NN	O	I-protein
comprising	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-DNA
lac	NN	O	I-DNA
repressor	NN	O	I-DNA
could	NN	O	O
be	NN	O	O
affinity-purified	NN	O	O
and	NN	O	O
recovered	NN	O	O
by	NN	O	O
gentle	NN	O	O
elution	NN	O	O
conditions	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
with	NN	O	O
a	NN	O	O
lactose	NN	O	O
analogue	NN	O	O
or	NN	O	O
by	NN	O	O
enzymatic	NN	O	O
means	NN	O	O
using	NN	O	O
either	NN	O	O
deoxyribonuclease	NN	O	B-protein
(	NN	O	O
DNase	NN	O	B-protein
)	NN	O	O
or	NN	O	O
restriction	NN	O	B-protein
endonucleases	NN	O	I-protein
.	NN	O	O

The	NN	O	O
results	NN	O	O
show	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
that	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
can	NN	O	O
be	NN	O	O
used	NN	O	O
for	NN	O	O
affinity	NN	O	O
purification	NN	O	O
of	NN	O	O
fusion	NN	O	B-protein
proteins	NN	O	I-protein
as	NN	O	O
exemplified	NN	O	O
by	NN	O	O
the	NN	O	O
specific	NN	O	O
and	NN	O	O
gentle	NN	O	O
recovery	NN	O	O
of	NN	O	O
beta-galactosidase	NN	O	B-protein
and	NN	O	O
alkaline	NN	O	B-protein
phosphatase	NN	O	I-protein
from	NN	O	O
bacterial	NN	O	O
lysates	NN	O	O
using	NN	O	O
immunomagnetic	NN	O	O
separation	NN	O	O
.	NN	O	O

The	NN	O	O
approach	NN	O	O
was	NN	O	O
further	NN	O	O
extended	NN	O	O
to	NN	O	O
cell	NN	O	O
separation	NN	O	O
by	NN	O	O
the	NN	O	O
efficient	NN	O	O
recovery	NN	O	O
and	NN	O	O
elution	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
CD37	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
.	NN	O	O

-DOCSTART-	O

ras	NN	O	B-protein
protein	NN	O	I-protein
activity	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
Jurkat	NN	O	B-cell_line
cell	NN	O	I-cell_line
model	NN	O	I-cell_line
of	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
an	NN	O	O
interleukin-2	NN	O	B-DNA
promoter/reporter	NN	O	I-DNA
gene	NN	O	I-DNA
construct	NN	O	I-DNA
was	NN	O	O
activated	NN	O	O
by	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
agonism	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
the	NN	O	O
lymphokine	NN	O	B-protein
interleukin-1	NN	O	B-protein
.	NN	O	O

Antigen	NN	O	O
receptor	NN	O	O
signals	NN	O	O
could	NN	O	O
be	NN	O	O
mimicked	NN	O	O
by	NN	O	O
suboptimal	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
calcium	NN	O	O
mobilization	NN	O	O
by	NN	O	O
an	NN	O	O
ionophore	NN	O	O
.	NN	O	O

In	NN	O	O
cotransfection	NN	O	O
experiments	NN	O	O
,	NN	O	O
oncogenic	NN	O	O
rats	NN	O	O
obviated	NN	O	O
the	NN	O	O
need	NN	O	O
for	NN	O	O
PKC	NN	O	B-protein
stimulation	NN	O	O
but	NN	O	O
did	NN	O	O
not	NN	O	O
replace	NN	O	O
either	NN	O	O
the	NN	O	O
calcium	NN	O	O
signal	NN	O	O
or	NN	O	O
interleukin-1	NN	O	B-protein
.	NN	O	O

Activated	NN	O	O
ras	NN	O	B-protein
expression	NN	O	O
also	NN	O	O
replaced	NN	O	O
the	NN	O	O
requirement	NN	O	O
for	NN	O	O
PKC	NN	O	B-protein
stimulation	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-AT	NN	O	B-protein
.	NN	O	O

A	NN	O	O
dominant	NN	O	B-protein
inhibitory	NN	O	I-protein
ras	NN	O	I-protein
mutant	NN	O	I-protein
specifically	NN	O	O
blocked	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
agonism	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
ras	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
signaling	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
blocked	NN	O	O
both	NN	O	O
activated	NN	O	O
ras	NN	O	B-protein
and	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
stimulation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
ras	NN	O	B-protein
upstream	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
Tat	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
HIV1	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Numerous	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
,	NN	O	O
upon	NN	O	O
HIV1	NN	O	O
infection	NN	O	O
,	NN	O	O
human	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
induced	NN	O	O
to	NN	O	O
differentiate	NN	O	O
,	NN	O	O
as	NN	O	O
indicated	NN	O	O
,	NN	O	O
for	NN	O	O
example	NN	O	O
,	NN	O	O
by	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	O
proteins	NN	O	O
involved	NN	O	O
in	NN	O	O
this	NN	O	O
process	NN	O	O
might	NN	O	O
be	NN	O	O
the	NN	O	O
Tat	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
this	NN	O	O
viral	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
productive	NN	O	O
infection	NN	O	O
,	NN	O	O
has	NN	O	O
also	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
display	NN	O	O
growth-stimulating	NN	O	O
properties	NN	O	O
and	NN	O	O
immunomodulatory	NN	O	O
activities	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
apprehend	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
HIV1	NN	O	B-DNA
tat	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
inducing	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HIV1-infected	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
have	NN	O	O
successfully	NN	O	O
introduced	NN	O	O
this	NN	O	O
gene	NN	O	O
into	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
infecting	NN	O	O
them	NN	O	O
with	NN	O	O
retroviral	NN	O	O
particles	NN	O	O
transducing	NN	O	O
tat	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
Tat	NN	O	B-protein
protein	NN	O	I-protein
constitutively	NN	O	O
expressed	NN	O	O
by	NN	O	O
these	NN	O	O
cells	NN	O	O
upon	NN	O	O
their	NN	O	O
differentiation	NN	O	O
was	NN	O	O
then	NN	O	O
evaluated	NN	O	O
by	NN	O	O
looking	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
of	NN	O	O
the	NN	O	O
c-fms	NN	O	B-DNA
proto-oncogenes	NN	O	I-DNA
which	NN	O	O
are	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
myelomonoblastic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
proto-oncogenes	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
this	NN	O	O
increase	NN	O	O
was	NN	O	O
amplified	NN	O	O
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
.	NN	O	O

No	NN	O	O
such	NN	O	O
increase	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
control	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
,	NN	O	O
among	NN	O	O
HIV1	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
,	NN	O	O
the	NN	O	O
Tat	NN	O	B-protein
protein	NN	O	I-protein
appears	NN	O	O
to	NN	O	O
trigger	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
viral	NN	O	B-protein
protein	NN	O	I-protein
directs	NN	O	O
progenitors	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
towards	NN	O	O
a	NN	O	O
differentiation	NN	O	O
stage	NN	O	O
in	NN	O	O
which	NN	O	O
production	NN	O	O
of	NN	O	O
viral	NN	O	B-protein
antigens	NN	O	I-protein
and	NN	O	O
virions	NN	O	O
might	NN	O	O
be	NN	O	O
more	NN	O	O
efficient	NN	O	O
.	NN	O	O

-DOCSTART-	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
preexisting	NN	O	O
and	NN	O	O
nuclear	NN	O	O
forms	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
.	NN	O	O

The	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
3	NN	O	O
is	NN	O	O
an	NN	O	O
inducible	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

NF-AT	NN	O	B-protein
is	NN	O	O
thought	NN	O	O
to	NN	O	O
consist	NN	O	O
of	NN	O	O
two	NN	O	O
components	NN	O	O
:	NN	O	O
a	NN	O	O
ubiquitous	NN	O	B-protein
,	NN	O	I-protein
inducible	NN	O	I-protein
nuclear	NN	O	I-protein
component	NN	O	I-protein
that	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
as	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
preexisting	NN	O	O
,	NN	O	O
T	NN	O	B-protein
cell-specific	NN	O	I-protein
component	NN	O	I-protein
(	NN	O	O
NF-ATp	NN	O	B-protein
)	NN	O	O
which	NN	O	O
is	NN	O	O
the	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
agents	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
and	NN	O	O
FK506	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
form	NN	O	O
two	NN	O	O
inducible	NN	O	O
NF-AT	NN	O	B-protein
complexes	NN	O	I-protein
with	NN	O	O
an	NN	O	O
oligonucleotide	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
NF-AT	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
IL-2	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
although	NN	O	O
hypotonic	NN	O	O
extracts	NN	O	O
of	NN	O	O
unstimulated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
form	NN	O	O
a	NN	O	O
single	NN	O	O
complex	NN	O	O
containing	NN	O	O
NF-ATp	NN	O	B-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
detect	NN	O	O
NF-ATp	NN	O	B-protein
in	NN	O	O
a	NN	O	O
gel	NN	O	O
shift	NN	O	O
assay	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
purification	NN	O	O
and	NN	O	O
biochemical	NN	O	O
studies	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
,	NN	O	O
is	NN	O	O
strikingly	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
precise	NN	O	O
sequence	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
oligonucleotide	NN	O	O
used	NN	O	O
as	NN	O	O
the	NN	O	O
labeled	NN	O	O
probe	NN	O	O
.	NN	O	O

Moreover	NN	O	O
we	NN	O	O
present	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
component	NN	O	O
that	NN	O	O
forms	NN	O	O
the	NN	O	O
faster-migrating	NN	O	O
(	NN	O	O
``	NN	O	O
lower	NN	O	O
''	NN	O	O
)	NN	O	O
nuclear	NN	O	B-protein
NF-AT	NN	O	I-protein
complex	NN	O	I-protein
is	NN	O	O
derived	NN	O	O
by	NN	O	O
a	NN	O	O
calcium-dependent	NN	O	O
,	NN	O	O
cyclosporin-sensitive	NN	O	O
,	NN	O	O
posttranslational	NN	O	O
modification	NN	O	O
of	NN	O	O
NF-ATp	NN	O	B-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
proteins	NN	O	O
stabilize	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
DNA	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
are	NN	O	O
discussed	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
a	NN	O	O
model	NN	O	O
relating	NN	O	O
the	NN	O	O
two	NN	O	O
nuclear	NN	O	O
NF-AT	NN	O	B-protein
complexes	NN	O	I-protein
to	NN	O	O
NF-ATp	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Cell	NN	O	O
cycle	NN	O	O
analysis	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
reveals	NN	O	O
novel	NN	O	B-protein
E2F	NN	O	I-protein
complexes	NN	O	I-protein
and	NN	O	O
biochemically	NN	O	O
distinct	NN	O	O
forms	NN	O	O
of	NN	O	O
free	NN	O	O
E2F	NN	O	B-protein
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
E2F	NN	O	B-protein
activates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
multiple	NN	O	O
genes	NN	O	B-DNA
involved	NN	O	O
in	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
c-myc	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
dihydrofolate	NN	O	B-DNA
reductase	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Regulation	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
involves	NN	O	O
its	NN	O	O
interactions	NN	O	O
with	NN	O	O
other	NN	O	O
cellular	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
including	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
Rb	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
Rb-related	NN	O	B-protein
protein	NN	O	I-protein
p107	NN	O	B-protein
,	NN	O	O
cyclin	NN	O	B-protein
A	NN	O	I-protein
,	NN	O	O
and	NN	O	O
cdk2	NN	O	B-protein
.	NN	O	O

We	NN	O	O
undertook	NN	O	O
a	NN	O	O
detailed	NN	O	O
analysis	NN	O	O
of	NN	O	O
E2F	NN	O	B-protein
DNA-binding	NN	O	O
activities	NN	O	O
and	NN	O	O
their	NN	O	O
cell	NN	O	O
cycle	NN	O	O
behavior	NN	O	O
in	NN	O	O
primary	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Three	NN	O	O
E2F	NN	O	B-protein
DNA-binding	NN	O	O
activities	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
resting	NN	O	O
(	NN	O	O
G0	NN	O	O
)	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
mobilities	NN	O	O
in	NN	O	O
gel	NN	O	O
shift	NN	O	O
assays	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
previously	NN	O	O
defined	NN	O	O
E2F	NN	O	B-protein
complexes	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
activities	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
novel	NN	O	O
,	NN	O	O
less	NN	O	O
abundant	NN	O	O
,	NN	O	O
Rb-	NN	O	O
E2F	NN	O	B-protein
complex	NN	O	O
.	NN	O	O

The	NN	O	O
most	NN	O	O
prominent	NN	O	O
E2F	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
resting	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
termed	NN	O	O
complex	NN	O	O
X	NN	O	O
)	NN	O	O
was	NN	O	O
abundant	NN	O	O
in	NN	O	O
both	NN	O	O
G0	NN	O	O
and	NN	O	O
G1	NN	O	O
but	NN	O	O
disappeared	NN	O	O
as	NN	O	O
cells	NN	O	O
entered	NN	O	O
S	NN	O	O
phase	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
possible	NN	O	O
role	NN	O	O
in	NN	O	O
negatively	NN	O	O
regulating	NN	O	O
E2F	NN	O	B-protein
function	NN	O	O
.	NN	O	O

Complex	NN	O	B-protein
X	NN	O	I-protein
could	NN	O	O
be	NN	O	O
dissociated	NN	O	O
by	NN	O	O
adenovirus	NN	O	O
E1A	NN	O	O
with	NN	O	O
a	NN	O	O
requirement	NN	O	O
for	NN	O	O
an	NN	O	O
intact	NN	O	O
E1A	NN	O	B-DNA
conserved	NN	O	I-DNA
region	NN	O	I-DNA
2	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
X	NN	O	B-protein
failed	NN	O	O
to	NN	O	O
react	NN	O	O
with	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
antibodies	NN	O	O
against	NN	O	O
Rb	NN	O	B-protein
or	NN	O	O
p107	NN	O	B-protein
,	NN	O	O
implicating	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
an	NN	O	O
E1A-binding	NN	O	B-protein
protein	NN	O	I-protein
other	NN	O	O
than	NN	O	O
Rb	NN	O	B-protein
or	NN	O	O
p107	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
these	NN	O	O
novel	NN	O	B-protein
E2F	NN	O	I-protein
complexes	NN	O	I-protein
,	NN	O	O
three	NN	O	O
distinct	NN	O	O
forms	NN	O	O
of	NN	O	O
unbound	NN	O	O
(	NN	O	O
free	NN	O	O
)	NN	O	O
E2F	NN	O	B-protein
were	NN	O	O
resolved	NN	O	O
in	NN	O	O
gel	NN	O	O
shift	NN	O	O
experiments	NN	O	O
.	NN	O	O

These	NN	O	O
species	NN	O	O
showed	NN	O	O
different	NN	O	O
cell	NN	O	O
cycle	NN	O	O
kinetics	NN	O	O
.	NN	O	O

UV	NN	O	O
cross-linking	NN	O	O
experiments	NN	O	O
suggested	NN	O	O
that	NN	O	O
a	NN	O	O
distinct	NN	O	O
E2F	NN	O	B-protein
DNA-binding	NN	O	O
protein	NN	O	O
is	NN	O	O
uniquely	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
S-phase	NN	O	B-protein
p107	NN	O	I-protein
complex	NN	O	I-protein
and	NN	O	O
is	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
Rb	NN	O	B-protein
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
E2F	NN	O	B-protein
consists	NN	O	O
of	NN	O	O
multiple	NN	O	O
,	NN	O	O
biochemically	NN	O	O
distinct	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
which	NN	O	O
function	NN	O	O
at	NN	O	O
different	NN	O	O
points	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

-DOCSTART-	O

Defective	NN	O	O
translocation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
in	NN	O	O
multidrug-resistant	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
confers	NN	O	O
a	NN	O	O
reversible	NN	O	O
loss	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
human	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
myeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
differentiate	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
.	NN	O	O

This	NN	O	O
event	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
c-jun	NN	O	B-DNA
early	NN	O	I-DNA
response	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
appearance	NN	O	O
of	NN	O	O
a	NN	O	O
monocytic	NN	O	O
phenotype	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
studies	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
vincristine-selected	NN	O	O
,	NN	O	O
multidrug	NN	O	O
resistance	NN	O	O
on	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
-induced	NN	O	O
HL-60	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
multidrug-resistant	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
designated	NN	O	O
HL-60/vinc	NN	O	O
,	NN	O	O
fail	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
TPA	NN	O	O
with	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
c-jun	NN	O	B-RNA
transcripts	NN	O	I-RNA
or	NN	O	O
other	NN	O	O
phenotypic	NN	O	O
characteristics	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

By	NN	O	O
contrast	NN	O	O
,	NN	O	O
treatment	NN	O	O
of	NN	O	O
HL-60/vinc	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
serine/threonine	NN	O	B-protein
protein	NN	O	I-protein
phosphatases	NN	O	I-protein
,	NN	O	O
induces	NN	O	O
c-jun	NN	O	B-DNA
transcription	NN	O	O
,	NN	O	O
growth	NN	O	O
arrest	NN	O	O
,	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
c-fms	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Studies	NN	O	O
were	NN	O	O
also	NN	O	O
performed	NN	O	O
with	NN	O	O
an	NN	O	O
HL-60/vinc	NN	O	B-cell_line
revertant	NN	O	I-cell_line
(	NN	O	O
HL-60/vinc/R	NN	O	B-cell_line
)	NN	O	O
line	NN	O	O
that	NN	O	O
has	NN	O	O
regained	NN	O	O
partial	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
vincristine	NN	O	O
.	NN	O	O

The	NN	O	O
finding	NN	O	O
that	NN	O	O
HL-60/vinc/R	NN	O	B-cell_line
cells	NN	O	I-cell_line
respond	NN	O	O
to	NN	O	O
TPA	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
monocytic	NN	O	O
phenotype	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
suggests	NN	O	O
that	NN	O	O
c-jun	NN	O	B-DNA
induction	NN	O	O
is	NN	O	O
not	NN	O	O
obligatory	NN	O	O
for	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Other	NN	O	O
studies	NN	O	O
further	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
jun-B	NN	O	B-DNA
and	NN	O	O
fra-1	NN	O	B-DNA
genes	NN	O	B-DNA
are	NN	O	O
induced	NN	O	O
by	NN	O	O
TPA	NN	O	O
in	NN	O	O
both	NN	O	O
HL-60/vinc	NN	O	B-cell_line
and	NN	O	O
HL-60/vinc/R	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
c-fos	NN	O	B-DNA
expression	NN	O	O
is	NN	O	O
attenuated	NN	O	O
in	NN	O	O
the	NN	O	O
HL-60/vinc	NN	O	O
line	NN	O	O
.	NN	O	O

Since	NN	O	O
TPA	NN	O	O
activates	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
translocation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
from	NN	O	O
the	NN	O	O
cytosol	NN	O	O
to	NN	O	O
the	NN	O	O
membrane	NN	O	O
fraction	NN	O	O
.	NN	O	O

Although	NN	O	O
HL-60	NN	O	B-cell_line
and	NN	O	O
HL-60/vinc/R	NN	O	B-cell_line
cells	NN	O	I-cell_line
demonstrated	NN	O	O
translocation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
this	NN	O	O
subcellular	NN	O	O
redistribution	NN	O	O
was	NN	O	O
undetectable	NN	O	O
in	NN	O	O
HL-60/vinc	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Activity	NN	O	O
of	NN	O	O
the	NN	O	O
mitogen-activated	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
family	NN	O	I-protein
with	NN	O	O
associated	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
Y-peptide	NN	O	I-protein
was	NN	O	O
markedly	NN	O	O
diminished	NN	O	O
in	NN	O	O
TPA-treated	NN	O	B-cell_line
HL-60/vinc	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
vincristine	NN	O	O
resistance	NN	O	O
confers	NN	O	O
insensitivity	NN	O	O
to	NN	O	O
TPA-induced	NN	O	O
differentiation	NN	O	O
and	NN	O	O
can	NN	O	O
include	NN	O	O
defects	NN	O	O
in	NN	O	O
PKC	NN	O	B-protein
-mediated	NN	O	O
signaling	NN	O	O
events	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
jun/fos	NN	O	B-DNA
early	NN	O	I-DNA
response	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
a	NN	O	O
multiple	NN	O	O
lymphokine	NN	O	B-protein
deficiency	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
a	NN	O	O
child	NN	O	O
with	NN	O	O
severe	NN	O	O
combined	NN	O	O
immunodeficiency	NN	O	O
are	NN	O	O
defective	NN	O	O
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
several	NN	O	O
lymphokine	NN	O	B-DNA
genes	NN	O	I-DNA
that	NN	O	O
include	NN	O	O
IL2	NN	O	B-DNA
,	NN	O	O
IL3	NN	O	B-DNA
,	NN	O	O
IL4	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
IL5	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
encode	NN	O	O
interleukins	NN	O	B-protein
2	NN	O	O
,	NN	O	O
3	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
and	NN	O	O
5	NN	O	O
(	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
-3	NN	O	B-protein
,	NN	O	O
-4	NN	O	B-protein
,	NN	O	O
and	NN	O	O
-5	NN	O	B-protein
)	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
defect	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
involved	NN	O	O
a	NN	O	O
trans-acting	NN	O	B-protein
factor	NN	O	I-protein
common	NN	O	O
to	NN	O	O
the	NN	O	O
affected	NN	O	O
lymphokine	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
from	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
to	NN	O	O
bind	NN	O	O
response	NN	O	O
elements	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
regulatory	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
IL2	NN	O	B-DNA
.	NN	O	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
NF-kB	NN	O	I-protein
,	NN	O	O
activation	NN	O	B-protein
protein	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
OCT-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-IL-2B	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
were	NN	O	O
normal	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
to	NN	O	O
its	NN	O	O
response	NN	O	O
element	NN	O	O
in	NN	O	O
the	NN	O	O
IL2	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
to	NN	O	O
an	NN	O	O
NF-AT-like	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
the	NN	O	O
IL4	NN	O	B-DNA
enhancer	NN	O	I-DNA
was	NN	O	O
abnormal	NN	O	O
.	NN	O	O

To	NN	O	O
ascertain	NN	O	O
whether	NN	O	O
the	NN	O	O
abnormal	NN	O	O
NF-AT	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
related	NN	O	O
to	NN	O	O
an	NN	O	O
impaired	NN	O	O
function	NN	O	O
,	NN	O	O
we	NN	O	O
transfected	NN	O	O
patient	NN	O	O
and	NN	O	O
control	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
constructs	NN	O	O
containing	NN	O	O
the	NN	O	O
reporter	NN	O	O
gene	NN	O	O
encoding	NN	O	O
chloramphenicol	NN	O	B-protein
acetyl	NN	O	I-protein
transferase	NN	O	I-protein
(	NN	O	O
CAT	NN	O	B-protein
)	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
the	NN	O	O
entire	NN	O	O
IL2	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
or	NN	O	O
of	NN	O	O
multimers	NN	O	O
of	NN	O	O
individual	NN	O	O
enhancer	NN	O	B-DNA
sequences	NN	O	I-DNA
.	NN	O	O

CAT	NN	O	O
expression	NN	O	O
directed	NN	O	O
by	NN	O	O
the	NN	O	O
IL2	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
or	NN	O	O
by	NN	O	O
a	NN	O	O
multimer	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT-binding	NN	O	B-DNA
site	NN	O	I-DNA
was	NN	O	O
markedly	NN	O	O
lower	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
relative	NN	O	O
to	NN	O	O
controls	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
CAT	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
directed	NN	O	O
by	NN	O	O
a	NN	O	O
multimer	NN	O	O
of	NN	O	O
the	NN	O	O
OCT-1	NN	O	B-DNA
proximal	NN	O	I-DNA
(	NN	O	I-DNA
OCT-1p	NN	O	I-DNA
)	NN	O	I-DNA
-binding	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
equivalent	NN	O	O
in	NN	O	O
patient	NN	O	O
and	NN	O	O
controls	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
an	NN	O	O
abnormality	NN	O	O
of/or	NN	O	O
influencing	NN	O	O
NF-AT	NN	O	B-protein
may	NN	O	O
underlie	NN	O	O
the	NN	O	O
multiple	NN	O	O
lymphokine	NN	O	B-protein
deficiency	NN	O	O
in	NN	O	O
this	NN	O	O
patient	NN	O	O
.	NN	O	O

-DOCSTART-	O

FK506	NN	O	O
and	NN	O	O
ciclosporin	NN	O	O
:	NN	O	O
molecular	NN	O	O
probes	NN	O	O
for	NN	O	O
studying	NN	O	O
intracellular	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

The	NN	O	O
immunosuppressants	NN	O	O
ciclosporin	NN	O	O
and	NN	O	O
FK506	NN	O	O
block	NN	O	O
the	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
signal-transduction	NN	O	O
pathway	NN	O	O
emanating	NN	O	O
from	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
thereby	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
helper	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
these	NN	O	O
drugs	NN	O	O
as	NN	O	O
probes	NN	O	O
,	NN	O	O
chemists	NN	O	O
and	NN	O	O
biologists	NN	O	O
have	NN	O	O
uncovered	NN	O	O
several	NN	O	O
intracellular	NN	O	O
signalling	NN	O	O
molecules	NN	O	O
bridging	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
second-messenger	NN	O	O
Ca2+	NN	O	O
ions	NN	O	O
and	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
among	NN	O	O
which	NN	O	O
are	NN	O	O
calmodulin	NN	O	B-protein
,	NN	O	O
calcineurin	NN	O	B-protein
and	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
Ca2+	NN	O	O
binds	NN	O	O
to	NN	O	O
calmodulin	NN	O	B-protein
,	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
calmodulin	NN	O	B-protein
to	NN	O	O
calcineurin	NN	O	B-protein
;	NN	O	O
the	NN	O	O
activated	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
in	NN	O	O
turn	NN	O	O
,	NN	O	O
may	NN	O	O
dephosphorylate	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	I-protein
NF-AT	NN	O	I-protein
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
its	NN	O	O
translocation	NN	O	O
from	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	NN	O	O
to	NN	O	O
form	NN	O	O
a	NN	O	O
competent	NN	O	B-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
.	NN	O	O

As	NN	O	O
described	NN	O	O
by	NN	O	O
Jun	NN	O	B-protein
Liu	NN	O	O
,	NN	O	O
these	NN	O	O
drugs	NN	O	O
manifest	NN	O	O
their	NN	O	O
effects	NN	O	O
in	NN	O	O
an	NN	O	O
unprecedented	NN	O	O
fashion	NN	O	O
.	NN	O	O

They	NN	O	O
do	NN	O	O
not	NN	O	O
directly	NN	O	O
intercept	NN	O	O
intracellular	NN	O	O
signalling	NN	O	O
molecules	NN	O	O
.	NN	O	O

Instead	NN	O	O
,	NN	O	O
they	NN	O	O
form	NN	O	O
tight	NN	O	O
complexes	NN	O	O
with	NN	O	O
two	NN	O	O
different	NN	O	O
classes	NN	O	O
of	NN	O	O
abundant	NN	O	O
cytosolic	NN	O	B-protein
receptors	NN	O	I-protein
called	NN	O	O
immunophilins	NN	O	B-protein
upon	NN	O	O
entering	NN	O	O
the	NN	O	O
cell	NN	O	O
,	NN	O	O
and	NN	O	O
consequently	NN	O	O
inhibit	NN	O	O
their	NN	O	O
peptidyl	NN	O	B-protein
prolyl	NN	O	I-protein
cis-trans	NN	O	I-protein
isomerase	NN	O	I-protein
activities	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
structurally	NN	O	O
distinct	NN	O	O
immunophilin-drug	NN	O	B-protein
complexes	NN	O	I-protein
bind	NN	O	O
to	NN	O	O
,	NN	O	O
and	NN	O	O
inhibit	NN	O	O
,	NN	O	O
the	NN	O	O
phosphatase	NN	O	B-protein
activity	NN	O	O
of	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

p105	NN	O	B-protein
and	NN	O	O
p98	NN	O	B-protein
precursor	NN	O	B-protein
proteins	NN	O	I-protein
play	NN	O	O
an	NN	O	O
active	NN	O	O
role	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

The	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
is	NN	O	O
composed	NN	O	O
of	NN	O	O
two	NN	O	O
distinct	NN	O	O
subgroups	NN	O	O
,	NN	O	O
proteins	NN	O	O
that	NN	O	O
undergo	NN	O	O
proteolytic	NN	O	O
processing	NN	O	O
and	NN	O	O
contain	NN	O	O
SWI6/ankyrin	NN	O	B-protein
repeats	NN	O	I-protein
in	NN	O	O
their	NN	O	O
carboxyl	NN	O	B-protein
termini	NN	O	I-protein
(	NN	O	O
p105	NN	O	B-protein
,	NN	O	O
p98	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
those	NN	O	O
without	NN	O	O
such	NN	O	O
repeats	NN	O	O
that	NN	O	O
do	NN	O	O
not	NN	O	O
require	NN	O	O
processing	NN	O	O
(	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
RelB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Dorsal	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
p105	NN	O	B-protein
and	NN	O	O
p98	NN	O	B-protein
precursors	NN	O	O
share	NN	O	O
functional	NN	O	O
properties	NN	O	O
with	NN	O	O
the	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
which	NN	O	O
also	NN	O	O
contain	NN	O	O
SWI6/ankyrin	NN	O	B-protein
repeats	NN	O	I-protein
.	NN	O	O

Both	NN	O	O
p105	NN	O	B-protein
and	NN	O	O
p98	NN	O	B-protein
were	NN	O	O
found	NN	O	O
to	NN	O	O
form	NN	O	O
stable	NN	O	O
complexes	NN	O	O
with	NN	O	O
other	NN	O	O
Rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
,	NN	O	O
including	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
c-Rel	NN	O	B-protein
.	NN	O	O

Association	NN	O	O
with	NN	O	O
the	NN	O	O
precursors	NN	O	O
is	NN	O	O
sufficient	NN	O	O
for	NN	O	O
cytoplasmic	NN	O	O
retention	NN	O	O
of	NN	O	O
either	NN	O	O
p65	NN	O	B-protein
or	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
otherwise	NN	O	O
nuclear	NN	O	O
.	NN	O	O

These	NN	O	O
complexes	NN	O	O
undergo	NN	O	O
stimulus-responsive	NN	O	O
processing	NN	O	O
to	NN	O	O
produce	NN	O	O
active	NN	O	O
p50/c-Rel	NN	O	B-protein
and	NN	O	O
p55/c-Rel	NN	O	B-protein
complexes	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
a	NN	O	O
second	NN	O	O
pathway	NN	O	O
leading	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
induction	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
processing	NN	O	O
of	NN	O	O
the	NN	O	O
precursors	NN	O	O
rather	NN	O	O
than	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
controls	NN	O	O
expression	NN	O	O
of	NN	O	O
inhibitor	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
:	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
inducible	NN	O	O
autoregulatory	NN	O	O
pathway	NN	O	O
.	NN	O	O

The	NN	O	O
eukaryotic	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor-kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
participates	NN	O	O
in	NN	O	O
many	NN	O	O
parts	NN	O	O
of	NN	O	O
the	NN	O	O
genetic	NN	O	O
program	NN	O	O
mediating	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
activation	NN	O	O
and	NN	O	O
growth	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
occurs	NN	O	O
after	NN	O	O
its	NN	O	O
induced	NN	O	O
dissociation	NN	O	O
from	NN	O	O
its	NN	O	O
cytoplasmic	NN	O	O
inhibitor	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

Phorbol	NN	O	O
ester	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
induction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
associated	NN	O	O
with	NN	O	O
both	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
performed	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Transfection	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
I	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
specifically	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
65-kilodalton	NN	O	B-protein
transactivating	NN	O	I-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Association	NN	O	O
of	NN	O	O
the	NN	O	O
newly	NN	O	O
synthesized	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
with	NN	O	O
p65	NN	O	B-protein
restores	NN	O	O
intracellular	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
prolongs	NN	O	O
the	NN	O	O
survival	NN	O	O
of	NN	O	O
this	NN	O	O
labile	NN	O	O
inhibitor	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
controls	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	O
autoregulatory	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
human	NN	O	B-DNA
prointerleukin	NN	O	I-DNA
1	NN	O	I-DNA
beta	NN	O	I-DNA
gene	NN	O	I-DNA
requires	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
both	NN	O	O
proximal	NN	O	O
and	NN	O	O
distal	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
for	NN	O	O
tissue-specific	NN	O	O
induction	NN	O	O
.	NN	O	O

In	NN	O	O
these	NN	O	O
studies	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
DNA	NN	O	B-DNA
sequences	NN	O	I-DNA
and	NN	O	O
specific	NN	O	O
protein	NN	O	O
interactions	NN	O	O
necessary	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
prointerleukin	NN	O	I-DNA
1	NN	O	I-DNA
beta	NN	O	I-DNA
(	NN	O	I-DNA
proIL-1	NN	O	I-DNA
beta	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
cell-type-independent	NN	O	B-DNA
lipopolysaccharide	NN	O	I-DNA
(	NN	O	I-DNA
LPS	NN	O	I-DNA
)	NN	O	I-DNA
-responsive	NN	O	I-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
located	NN	O	O
between	NN	O	O
-3757	NN	O	B-DNA
and	NN	O	I-DNA
-2729	NN	O	I-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
cap	NN	O	B-DNA
site	NN	O	I-DNA
)	NN	O	O
consisted	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
six	NN	O	O
discrete	NN	O	O
subregions	NN	O	O
which	NN	O	O
were	NN	O	O
essential	NN	O	O
to	NN	O	O
the	NN	O	O
maximal	NN	O	O
induction	NN	O	O
by	NN	O	O
LPS	NN	O	O
in	NN	O	O
transfected	NN	O	B-cell_line
monocytes	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
enhancer	NN	O	O
also	NN	O	O
appeared	NN	O	O
to	NN	O	O
mediate	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
induction	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
IL-1	NN	O	B-protein
responsiveness	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

Deletion	NN	O	O
and	NN	O	O
base	NN	O	O
substitution	NN	O	O
mutations	NN	O	O
along	NN	O	O
with	NN	O	O
DNA	NN	O	O
binding	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
enhancer	NN	O	O
contained	NN	O	O
a	NN	O	O
minimum	NN	O	O
of	NN	O	O
three	NN	O	O
functional	NN	O	O
protein	NN	O	B-DNA
binding	NN	O	I-DNA
sequences	NN	O	I-DNA
,	NN	O	O
two	NN	O	O
of	NN	O	O
which	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
gene	NN	O	O
induction	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
essential	NN	O	O
proteins	NN	O	O
which	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
enhancer	NN	O	O
was	NN	O	O
similar	NN	O	O
or	NN	O	O
identical	NN	O	O
to	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
C/EBP	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
required	NN	O	O
for	NN	O	O
both	NN	O	O
IL-1	NN	O	B-protein
-and	NN	O	O
LPS-specific	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
gene	NN	O	I-DNA
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
the	NN	O	O
NF-IL6	NN	O	B-DNA
proteins	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

When	NN	O	O
ligated	NN	O	O
to	NN	O	O
the	NN	O	O
proIL-1	NN	O	B-DNA
beta	NN	O	I-DNA
cap	NN	O	I-DNA
site	NN	O	I-DNA
-proximal	NN	O	B-DNA
region	NN	O	I-DNA
(	NN	O	O
located	NN	O	O
between	NN	O	O
-131	NN	O	O
to	NN	O	O
+12	NN	O	O
)	NN	O	O
,	NN	O	O
both	NN	O	O
the	NN	O	O
proIL-1	NN	O	B-DNA
beta	NN	O	I-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
simian	NN	O	I-DNA
virus	NN	O	I-DNA
40	NN	O	I-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
functioned	NN	O	O
more	NN	O	O
efficiently	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
than	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
are	NN	O	O
not	NN	O	O
normally	NN	O	O
competent	NN	O	O
for	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

When	NN	O	O
ligated	NN	O	O
to	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
c-fos	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
proIL-1	NN	O	B-DNA
beta	NN	O	I-DNA
enhancer	NN	O	I-DNA
was	NN	O	O
inducible	NN	O	O
in	NN	O	O
phorbol	NN	O	B-cell_line
myristate	NN	O	I-cell_line
acetate-stimulated	NN	O	I-cell_line
HeLa	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
proIL-1	NN	O	B-protein
beta	NN	O	I-protein
promoter-proximal	NN	O	O
requirement	NN	O	O
for	NN	O	O
tissue	NN	O	O
specificity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lymphocytes	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
disease	NN	O	O
suggest	NN	O	O
adenovirus	NN	O	O
is	NN	O	O
one	NN	O	O
cause	NN	O	O
of	NN	O	O
persistent	NN	O	O
or	NN	O	O
recurrent	NN	O	O
inflammatory	NN	O	O
arthritis	NN	O	O
.	NN	O	O

The	NN	O	O
assessment	NN	O	O
of	NN	O	O
synovial	NN	O	O
lymphocyte	NN	O	O
reactivity	NN	O	O
to	NN	O	O
adenovirus	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
was	NN	O	O
undertaken	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
persistent	NN	O	O
or	NN	O	O
recurrent	NN	O	O
inflammatory	NN	O	O
arthritis	NN	O	O
.	NN	O	O

The	NN	O	O
3H-thymidine	NN	O	O
uptake	NN	O	O
procedure	NN	O	O
was	NN	O	O
employed	NN	O	O
,	NN	O	O
incorporating	NN	O	O
multiple	NN	O	O
microbiological	NN	O	B-protein
antigens	NN	O	I-protein
.	NN	O	O

Five	NN	O	O
patients	NN	O	O
were	NN	O	O
found	NN	O	O
with	NN	O	O
repeated	NN	O	O
maximal	NN	O	O
responses	NN	O	O
to	NN	O	O
adenovirus	NN	O	B-protein
antigen	NN	O	I-protein
;	NN	O	O
in	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
adenovirus	NN	O	B-DNA
nucleotide	NN	O	I-DNA
sequences	NN	O	I-DNA
were	NN	O	O
present	NN	O	O
in	NN	O	O
a	NN	O	O
synovial	NN	O	O
biopsy	NN	O	O
specimen	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
adenovirus	NN	O	O
may	NN	O	O
be	NN	O	O
one	NN	O	O
cause	NN	O	O
of	NN	O	O
persistent	NN	O	O
or	NN	O	O
recurrent	NN	O	O
inflammatory	NN	O	O
arthritis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
PILOT	NN	O	B-protein
,	NN	O	O
a	NN	O	O
putative	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
requires	NN	O	O
two	NN	O	O
signals	NN	O	O
and	NN	O	O
is	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
sensitive	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Few	NN	O	O
known	NN	O	O
genes	NN	O	B-DNA
(	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
IL-8	NN	O	B-DNA
family	NN	O	I-DNA
,	NN	O	O
interferon-gamma	NN	O	B-DNA
)	NN	O	O
are	NN	O	O
induced	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
only	NN	O	O
through	NN	O	O
the	NN	O	O
combined	NN	O	O
effect	NN	O	O
of	NN	O	O
phorbol	NN	O	O
myristic	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-ionophore	NN	O	O
,	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
only	NN	O	O
these	NN	O	O
genes	NN	O	B-DNA
can	NN	O	O
be	NN	O	O
fully	NN	O	O
suppressed	NN	O	O
by	NN	O	O
Cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CyA	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
putative	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
designated	NN	O	O
PILOT	NN	O	B-protein
,	NN	O	O
with	NN	O	O
an	NN	O	O
identical	NN	O	O
dual	NN	O	O
signal	NN	O	O
requirement	NN	O	O
for	NN	O	O
expression	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
the	NN	O	O
PILOT	NN	O	B-protein
gene	NN	O	O
is	NN	O	O
detectable	NN	O	O
in	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
20	NN	O	O
min	NN	O	O
following	NN	O	O
activation	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cycloheximide	NN	O	O
and	NN	O	O
is	NN	O	O
fully	NN	O	O
suppressed	NN	O	O
by	NN	O	O
CyA	NN	O	O
.	NN	O	O

The	NN	O	O
PILOT	NN	O	B-protein
protein	NN	O	O
has	NN	O	O
a	NN	O	O
calculated	NN	O	O
M	NN	O	O
(	NN	O	O
r	NN	O	O
)	NN	O	O
of	NN	O	O
42.6	NN	O	O
kDa	NN	O	O
and	NN	O	O
contains	NN	O	O
three	NN	O	O
zinc	NN	O	B-protein
fingers	NN	O	I-protein
of	NN	O	O
the	NN	O	O
C2H2-type	NN	O	B-protein
at	NN	O	O
the	NN	O	O
carboxyl-terminus	NN	O	B-protein
which	NN	O	O
are	NN	O	O
highly	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
regions	NN	O	I-protein
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
EGR1	NN	O	B-protein
,	NN	O	O
EGR2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
pAT	NN	O	B-protein
133	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
PILOT	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
requires	NN	O	O
only	NN	O	O
one	NN	O	O
signal	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
CyA	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
directly	NN	O	O
demonstrates	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
Ca2+	NN	O	B-protein
signal-dependent	NN	O	I-protein
regulatory	NN	O	I-protein
element	NN	O	I-protein
obligatory	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
some	NN	O	O
genes	NN	O	B-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

This	NN	O	O
differential	NN	O	O
expression	NN	O	O
model	NN	O	O
will	NN	O	O
be	NN	O	O
valuable	NN	O	O
in	NN	O	O
the	NN	O	O
dissection	NN	O	O
of	NN	O	O
the	NN	O	O
dual	NN	O	O
signal	NN	O	O
pathway	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
CyA	NN	O	O
upon	NN	O	O
it	NN	O	O
.	NN	O	O

-DOCSTART-	O

Antisense	NN	O	O
oligonucleotides	NN	O	O
to	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
block	NN	O	O
CD11b	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
alter	NN	O	O
adhesion	NN	O	O
properties	NN	O	O
of	NN	O	O
differentiated	NN	O	O
HL-60	NN	O	B-cell_line
granulocytes	NN	O	I-cell_line
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
pleiotropic	NN	O	B-protein
regulator	NN	O	I-protein
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
genes	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
response	NN	O	O
to	NN	O	O
injury	NN	O	O
.	NN	O	O

This	NN	O	O
function	NN	O	O
has	NN	O	O
been	NN	O	O
attributed	NN	O	O
to	NN	O	O
the	NN	O	O
coordinated	NN	O	O
binding	NN	O	O
of	NN	O	O
subunits	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
to	NN	O	O
distinct	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
numerous	NN	O	O
genes	NN	O	O
,	NN	O	O
including	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
growth	NN	O	B-protein
factor	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

Antisense	NN	O	O
phosphorothioate	NN	O	O
oligonucleotides	NN	O	O
to	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
subunits	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
were	NN	O	O
used	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
physiologic	NN	O	O
role	NN	O	O
of	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
resting	NN	O	B-cell_line
and	NN	O	I-cell_line
stimulated	NN	O	I-cell_line
granulocytes	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
was	NN	O	O
produced	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
the	NN	O	O
phosphorothioate	NN	O	O
antisense	NN	O	O
oligodeoxynucleotide	NN	O	O
.	NN	O	O

This	NN	O	O
reduction	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
rapid	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
adhesion	NN	O	O
of	NN	O	O
dimethyl	NN	O	B-cell_line
sulfoxide-differentiated	NN	O	I-cell_line
HL-60	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
stimulated	NN	O	O
by	NN	O	O
12-O-tetradecanoylphorbol	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
effects	NN	O	O
were	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
marked	NN	O	O
reduction	NN	O	O
in	NN	O	O
CD11b	NN	O	B-protein
integrin	NN	O	B-protein
expression	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
antisense	NN	O	O
oligomer	NN	O	O
effectively	NN	O	O
abolished	NN	O	O
an	NN	O	O
upregulation	NN	O	O
of	NN	O	O
CD11b	NN	O	B-protein
that	NN	O	O
was	NN	O	O
produced	NN	O	O
by	NN	O	O
formyl-met-leu-phe	NN	O	O
and	NN	O	O
TPA	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
antisense	NN	O	O
phosphorothioate	NN	O	O
oligodeoxynucleotide	NN	O	O
had	NN	O	O
no	NN	O	O
significant	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
or	NN	O	O
on	NN	O	O
phagocytosis	NN	O	O
by	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
antisense	NN	O	O
oligomers	NN	O	O
to	NN	O	O
p65	NN	O	B-protein
can	NN	O	O
be	NN	O	O
used	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
activation	NN	O	O
pathways	NN	O	O
of	NN	O	O
neutrophils	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
p65	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
regulates	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
by	NN	O	O
two	NN	O	O
distinct	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

Transcription	NN	O	O
factor	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
p50/p65	NN	O	B-protein
)	NN	O	O
is	NN	O	O
generally	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
by	NN	O	O
its	NN	O	O
inhibitor	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Overproduced	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
free	NN	O	O
from	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
is	NN	O	O
rapidly	NN	O	O
degraded	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
increases	NN	O	O
endogenous	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
protein	NN	O	O
in	NN	O	O
both	NN	O	O
carcinoma	NN	O	B-cell_type
and	NN	O	I-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
two	NN	O	O
mechanisms	NN	O	O
:	NN	O	O
protein	NN	O	O
stabilization	NN	O	O
and	NN	O	O
increased	NN	O	O
transcription	NN	O	O
of	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
p65	NN	O	B-protein
delta	NN	O	I-protein
,	NN	O	O
a	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
splice	NN	O	O
variant	NN	O	O
,	NN	O	O
fails	NN	O	O
to	NN	O	O
markedly	NN	O	O
augment	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
protein	NN	O	O
levels	NN	O	O
.	NN	O	O

Both	NN	O	O
overexpressed	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
coexpressed	NN	O	O
p50	NN	O	B-protein
are	NN	O	O
cytoplasmic	NN	O	O
,	NN	O	O
whereas	NN	O	O
p65	NN	O	B-protein
delta	NN	O	I-protein
is	NN	O	O
partly	NN	O	O
nuclear	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
p65	NN	O	B-protein
can	NN	O	O
maintain	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
are	NN	O	O
linked	NN	O	O
in	NN	O	O
an	NN	O	O
autoregulatory	NN	O	O
loop	NN	O	O
,	NN	O	O
ensuring	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
held	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
until	NN	O	O
cells	NN	O	O
are	NN	O	O
specifically	NN	O	O
induced	NN	O	O
to	NN	O	O
translocate	NN	O	O
it	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lipopolysaccharide	NN	O	O
induces	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
MAD3	NN	O	B-protein
and	NN	O	O
activation	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
and	NN	O	O
related	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
proteins	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Many	NN	O	O
effects	NN	O	O
of	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
on	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
including	NN	O	O
that	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
,	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
kappa	NN	O	O
B	NN	O	O
DNA-binding	NN	O	O
proteins	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
specific	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
LPS	NN	O	O
response	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
through	NN	O	O
which	NN	O	O
LPS-generated	NN	O	O
signals	NN	O	O
are	NN	O	O
transduced	NN	O	O
remain	NN	O	O
unclear	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
LPS	NN	O	O
induces	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
c-Rel/p50	NN	O	B-protein
heterodimers	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
p50/p65	NN	O	B-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
kappa	NN	O	O
B	NN	O	O
DNA-binding	NN	O	O
complexes	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
occurred	NN	O	O
concomitantly	NN	O	O
with	NN	O	O
a	NN	O	O
rapid	NN	O	O
decrease	NN	O	O
in	NN	O	O
their	NN	O	O
cytosolic	NN	O	O
levels	NN	O	O
and	NN	O	O
was	NN	O	O
independent	NN	O	O
of	NN	O	O
phorbol	NN	O	B-protein
ester-sensitive	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
.	NN	O	O

Within	NN	O	O
24	NN	O	O
h	NN	O	O
following	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
striking	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
p105	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p50	NN	O	B-protein
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
.	NN	O	O

The	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
correlated	NN	O	O
with	NN	O	O
increases	NN	O	O
in	NN	O	O
the	NN	O	O
amounts	NN	O	O
of	NN	O	O
their	NN	O	O
mRNAs	NN	O	B-RNA
during	NN	O	O
LPS	NN	O	O
activation	NN	O	O
of	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

LPS	NN	O	O
activation	NN	O	O
of	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
resulted	NN	O	O
in	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
MAD3	NN	O	B-protein
(	NN	O	O
an	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
-like	NN	O	O
protein	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
rapid	NN	O	O
increase	NN	O	O
in	NN	O	O
MAD3	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
MAD3	NN	O	B-protein
protein	NN	O	O
by	NN	O	O
2	NN	O	O
h	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
LPS	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
results	NN	O	O
in	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
c-Rel/p50	NN	O	B-protein
and	NN	O	O
p50/p65	NN	O	B-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
and	NN	O	O
induces	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
MAD3	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
chimeric	NN	O	B-protein
type	NN	O	I-protein
II/type	NN	O	I-protein
I	NN	O	I-protein
interleukin-1	NN	O	I-protein
receptor	NN	O	I-protein
can	NN	O	O
mediate	NN	O	O
interleukin-1	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
interleukin-1	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-1R	NN	O	B-protein
)	NN	O	O
is	NN	O	O
capable	NN	O	O
of	NN	O	O
transducing	NN	O	O
a	NN	O	O
signal	NN	O	O
resulting	NN	O	O
in	NN	O	O
promoter	NN	O	O
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
signal	NN	O	O
transduction	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
which	NN	O	O
consists	NN	O	O
of	NN	O	O
213	NN	O	O
amino	NN	O	O
acids	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
the	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
IL-1R	NN	O	I-protein
has	NN	O	O
a	NN	O	O
small	NN	O	O
cytoplasmic	NN	O	B-protein
tail	NN	O	I-protein
,	NN	O	O
and	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
clear	NN	O	O
whether	NN	O	O
this	NN	O	O
receptor	NN	O	O
is	NN	O	O
a	NN	O	O
signal-transducing	NN	O	B-protein
or	NN	O	O
a	NN	O	O
regulatory	NN	O	B-protein
molecule	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
the	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
IL-1R	NN	O	I-protein
does	NN	O	O
not	NN	O	O
mediate	NN	O	O
gene	NN	O	O
activation	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
a	NN	O	O
hybrid	NN	O	O
receptor	NN	O	O
composed	NN	O	O
of	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
and	NN	O	I-protein
transmembrane	NN	O	I-protein
regions	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
type	NN	O	I-protein
II	NN	O	I-protein
interleukin-1	NN	O	I-protein
fused	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
type	NN	O	I-protein
I	NN	O	I-protein
IL-1R	NN	O	I-protein
was	NN	O	O
capable	NN	O	O
of	NN	O	O
transducing	NN	O	O
a	NN	O	O
signal	NN	O	O
across	NN	O	O
the	NN	O	O
membrane	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
pattern	NN	O	O
of	NN	O	O
gene	NN	O	O
activation	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
IL-1R	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
extracellular	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
IL-1R	NN	O	I-protein
was	NN	O	O
capable	NN	O	O
of	NN	O	O
functionally	NN	O	O
interacting	NN	O	O
with	NN	O	O
interleukin-1	NN	O	B-protein
and	NN	O	O
transmitting	NN	O	O
the	NN	O	O
resulting	NN	O	O
signal	NN	O	O
to	NN	O	O
a	NN	O	O
heterologous	NN	O	B-protein
cytoplasmic	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
primary	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
.	NN	O	O

The	NN	O	O
precise	NN	O	O
molecular	NN	O	O
abnormalities	NN	O	O
that	NN	O	O
cause	NN	O	O
primary	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
completely	NN	O	O
described	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
subject	NN	O	O
with	NN	O	O
primary	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
we	NN	O	O
have	NN	O	O
observed	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
hGR	NN	O	B-protein
)	NN	O	O
with	NN	O	O
a	NN	O	O
decreased	NN	O	O
affinity	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

We	NN	O	O
hypothesize	NN	O	O
that	NN	O	O
a	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
hGR	NN	O	B-protein
glucocorticoid-binding	NN	O	I-protein
domain	NN	O	I-protein
is	NN	O	O
the	NN	O	O
cause	NN	O	O
of	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
.	NN	O	O

Total	NN	O	O
RNA	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
index	NN	O	O
subject	NN	O	O
's	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
was	NN	O	O
used	NN	O	O
to	NN	O	O
produce	NN	O	O
first	NN	O	O
strand	NN	O	O
hGR	NN	O	B-DNA
cDNAs	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
entire	NN	O	O
hGR	NN	O	B-DNA
cDNA	NN	O	I-DNA
was	NN	O	O
amplified	NN	O	O
in	NN	O	O
segments	NN	O	O
and	NN	O	O
sequenced	NN	O	O
.	NN	O	O

At	NN	O	O
nucleotide	NN	O	O
2	NN	O	O
,	NN	O	O
317	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
homozygous	NN	O	O
A	NN	O	O
for	NN	O	O
G	NN	O	O
point	NN	O	O
mutation	NN	O	O
that	NN	O	O
predicts	NN	O	O
an	NN	O	O
isoleucine	NN	O	O
(	NN	O	O
ATT	NN	O	O
)	NN	O	O
for	NN	O	O
valine	NN	O	O
(	NN	O	O
GTT	NN	O	O
)	NN	O	O
substitution	NN	O	O
at	NN	O	O
amino	NN	O	O
acid	NN	O	O
729	NN	O	O
.	NN	O	O

When	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
hGR	NN	O	I-protein
and	NN	O	O
hGR-Ile	NN	O	B-protein
729	NN	O	I-protein
were	NN	O	O
expressed	NN	O	O
in	NN	O	O
COS-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
assayed	NN	O	O
for	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-Dexamethasone	NN	O	O
binding	NN	O	O
,	NN	O	O
the	NN	O	O
dissociation	NN	O	O
constants	NN	O	O
were	NN	O	O
0.799	NN	O	O
+/-	NN	O	O
0.068	NN	O	O
and	NN	O	O
1.54	NN	O	O
+/-	NN	O	O
0.06	NN	O	O
nM	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
)	NN	O	O
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

When	NN	O	O
the	NN	O	O
wild-type	NN	O	B-protein
hGR	NN	O	I-protein
and	NN	O	O
hGR-Ile	NN	O	B-protein
729	NN	O	I-protein
were	NN	O	O
expressed	NN	O	O
in	NN	O	O
CV-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
were	NN	O	O
cotransfected	NN	O	O
with	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
mammary	NN	O	I-DNA
tumor	NN	O	I-DNA
virus	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
fused	NN	O	O
to	NN	O	O
the	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyl	NN	O	I-DNA
transferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
hGR-Ile	NN	O	B-protein
729	NN	O	I-protein
conferred	NN	O	O
a	NN	O	O
fourfold	NN	O	O
decrease	NN	O	O
in	NN	O	O
apparent	NN	O	O
potency	NN	O	O
on	NN	O	O
dexamethasone	NN	O	O
stimulation	NN	O	O
of	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

The	NN	O	O
isoleucine	NN	O	O
for	NN	O	O
valine	NN	O	O
substitution	NN	O	O
at	NN	O	O
amino	NN	O	O
acid	NN	O	O
729	NN	O	O
impairs	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
hGR	NN	O	B-protein
and	NN	O	O
is	NN	O	O
the	NN	O	O
likely	NN	O	O
cause	NN	O	O
of	NN	O	O
primary	NN	O	O
cortisol	NN	O	O
resistance	NN	O	O
in	NN	O	O
this	NN	O	O
subject	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
transcriptionally	NN	O	B-protein
active	NN	O	I-protein
factors	NN	O	I-protein
mediating	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-protein
Tax	NN	O	I-protein
transactivator	NN	O	I-protein
on	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
include	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
c-rel	NN	O	B-DNA
proto-oncogene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
transactivator	NN	O	O
HTLV-I	NN	O	B-protein
Tax	NN	O	I-protein
activates	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
the	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
alpha-chain	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
via	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
that	NN	O	O
can	NN	O	O
bind	NN	O	O
several	NN	O	O
protein	NN	O	O
species	NN	O	O
of	NN	O	O
the	NN	O	O
rel	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

Tax1	NN	O	B-protein
strongly	NN	O	O
activates	NN	O	O
the	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
motif	NN	O	O
,	NN	O	O
in	NN	O	O
both	NN	O	O
epithelial	NN	O	B-cell_line
HeLa	NN	O	I-cell_line
and	NN	O	O
lymphoid	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
activation	NN	O	O
was	NN	O	O
not	NN	O	O
observed	NN	O	O
in	NN	O	O
undifferentiated	NN	O	O
embryocarcinoma	NN	O	O
F9	NN	O	O
cells	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
Rel	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
showed	NN	O	O
that	NN	O	O
significant	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
observed	NN	O	O
only	NN	O	O
with	NN	O	O
Rel	NN	O	B-protein
and	NN	O	O
Rel	NN	O	B-protein
plus	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
whereas	NN	O	O
both	NN	O	O
Tax	NN	O	B-protein
and	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
are	NN	O	O
able	NN	O	O
to	NN	O	O
efficiently	NN	O	O
induce	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
to	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
PMA	NN	O	O
is	NN	O	O
functionally	NN	O	O
inactive	NN	O	O
.	NN	O	O

Using	NN	O	O
the	NN	O	O
DNA	NN	O	O
affinity	NN	O	O
precipitation	NN	O	O
assay	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
Tax1	NN	O	B-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
efficiently	NN	O	O
induce	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
Rel	NN	O	O
,	NN	O	O
whereas	NN	O	O
PMA	NN	O	O
is	NN	O	O
not	NN	O	O
.	NN	O	O

This	NN	O	O
established	NN	O	O
a	NN	O	O
clear	NN	O	O
difference	NN	O	O
between	NN	O	O
both	NN	O	O
stimuli	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
Rel	NN	O	B-protein
is	NN	O	O
the	NN	O	O
functionally	NN	O	B-protein
active	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
from	NN	O	O
these	NN	O	O
results	NN	O	O
that	NN	O	O
the	NN	O	O
functional	NN	O	O
activity	NN	O	O
of	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
rel	NN	O	B-protein
family	NN	O	I-protein
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
their	NN	O	O
interaction	NN	O	O
with	NN	O	O
DNA	NN	O	O
and	NN	O	O
that	NN	O	O
Rel	NN	O	B-protein
can	NN	O	O
be	NN	O	O
a	NN	O	O
potent	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
on	NN	O	O
specific	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
3	NN	O	I-protein
(	NN	O	O
IL3	NN	O	B-protein
)	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Role	NN	O	O
of	NN	O	O
AP-1-	NN	O	B-protein
and	NN	O	I-protein
octamer-binding	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
control	NN	O	O
of	NN	O	O
IL3	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
molecular	NN	O	O
and	NN	O	O
biochemical	NN	O	O
basis	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
3	NN	O	I-protein
(	NN	O	O
IL3	NN	O	B-protein
)	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
following	NN	O	O
CD3	NN	O	O
and	NN	O	O
CD2	NN	O	O
receptor	NN	O	O
stimulation	NN	O	O
or	NN	O	O
activation	NN	O	O
by	NN	O	O
phytohemagglutinin	NN	O	B-protein
plus	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
.	NN	O	O

Using	NN	O	O
transfection	NN	O	O
and	NN	O	O
reporter	NN	O	O
gene	NN	O	O
assays	NN	O	O
specifically	NN	O	O
designed	NN	O	O
for	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
gel	NN	O	O
retardation	NN	O	O
assays	NN	O	O
,	NN	O	O
Western	NN	O	O
blot	NN	O	O
analyses	NN	O	O
and	NN	O	O
UV	NN	O	O
cross-linking	NN	O	O
studies	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
octamer-binding	NN	O	B-protein
proteins	NN	O	I-protein
play	NN	O	O
a	NN	O	O
major	NN	O	O
role	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL3	NN	O	B-DNA
gene	NN	O	I-DNA
via	NN	O	O
their	NN	O	O
interaction	NN	O	O
with	NN	O	O
two	NN	O	O
specific	NN	O	O
regions	NN	O	O
contained	NN	O	O
within	NN	O	O
the	NN	O	O
IL3	NN	O	B-DNA
5'-flanking	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
the	NN	O	O
region	NN	O	O
between	NN	O	O
bases	NN	O	O
-107	NN	O	B-DNA
and	NN	O	I-DNA
-59	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL3	NN	O	B-DNA
promoter	NN	O	I-DNA
containing	NN	O	O
putative	NN	O	O
AP-2	NN	O	B-DNA
and	NN	O	I-DNA
Sp1	NN	O	I-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
appears	NN	O	O
necessary	NN	O	O
for	NN	O	O
basal	NN	O	O
level	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL3	NN	O	B-protein
gene	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
also	NN	O	O
indicate	NN	O	O
that	NN	O	O
CD2	NN	O	B-protein
receptor	NN	O	O
activation	NN	O	O
and	NN	O	O
phytohemagglutinin	NN	O	B-protein
plus	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
stimulation	NN	O	O
augment	NN	O	O
T	NN	O	O
cell	NN	O	O
IL3	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
through	NN	O	O
the	NN	O	O
same	NN	O	O
cis-	NN	O	O
and	NN	O	O
trans-activating	NN	O	O
signals	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
should	NN	O	O
contribute	NN	O	O
to	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
IL3	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Dimerization	NN	O	O
of	NN	O	O
NF-KB2	NN	O	B-protein
with	NN	O	O
RelA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
regulates	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
inhibition	NN	O	O
by	NN	O	O
an	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
(	NN	O	O
MAD-3	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
a	NN	O	O
cellular	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
composed	NN	O	O
of	NN	O	O
distinct	NN	O	O
subunits	NN	O	O
;	NN	O	O
five	NN	O	O
independent	NN	O	O
genes	NN	O	O
,	NN	O	O
NFKB1	NN	O	B-DNA
(	NN	O	O
p105	NN	O	B-protein
)	NN	O	O
,	NN	O	O
NFKB2	NN	O	B-DNA
(	NN	O	O
p100	NN	O	B-protein
)	NN	O	O
,	NN	O	O
RelA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
,	NN	O	O
c-rel	NN	O	B-DNA
and	NN	O	O
relB	NN	O	B-DNA
,	NN	O	O
that	NN	O	O
encode	NN	O	O
related	NN	O	O
proteins	NN	O	O
that	NN	O	O
bind	NN	O	O
to	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
DNA	NN	O	I-DNA
elements	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
isolated	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
found	NN	O	O
that	NN	O	O
NFKB2	NN	O	B-protein
(	NN	O	I-protein
p49/p52	NN	O	I-protein
)	NN	O	I-protein
acts	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
RelA	NN	O	B-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
to	NN	O	O
stimulate	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	O
T-leukemia	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Here	NN	O	O
we	NN	O	O
examine	NN	O	O
the	NN	O	O
biochemical	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
HIV	NN	O	O
by	NN	O	O
NFKB2	NN	O	B-protein
.	NN	O	O

Using	NN	O	O
Scatchard	NN	O	O
analysis	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
determined	NN	O	O
the	NN	O	O
dissociation	NN	O	O
constants	NN	O	O
of	NN	O	O
homodimeric	NN	O	B-protein
p49	NN	O	I-protein
and	NN	O	O
heterodimeric	NN	O	B-protein
p49/p65	NN	O	I-protein
for	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

p49	NN	O	B-protein
has	NN	O	O
a	NN	O	O
approximately	NN	O	O
18-fold-lower	NN	O	O
affinity	NN	O	O
for	NN	O	O
the	NN	O	O
HIV	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
KD	NN	O	O
=	NN	O	O
69.1	NN	O	O
pM	NN	O	O
)	NN	O	O
than	NN	O	O
does	NN	O	O
the	NN	O	O
approximately	NN	O	O
50-kDa	NN	O	O
protein	NN	O	O
NFKB1	NN	O	B-protein
(	NN	O	I-protein
p50	NN	O	I-protein
)	NN	O	I-protein
derived	NN	O	O
from	NN	O	O
p105	NN	O	B-protein
(	NN	O	O
KD	NN	O	O
=	NN	O	O
3.9	NN	O	O
pM	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
affinity	NN	O	O
of	NN	O	O
heterodimeric	NN	O	B-protein
NFKB2	NN	O	I-protein
(	NN	O	I-protein
p49	NN	O	I-protein
)	NN	O	I-protein
/RelA	NN	O	I-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
for	NN	O	O
this	NN	O	O
site	NN	O	O
is	NN	O	O
approximately	NN	O	O
6-fold	NN	O	O
higher	NN	O	O
(	NN	O	O
KD	NN	O	O
=	NN	O	O
11.8	NN	O	O
pM	NN	O	O
)	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
p49	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
these	NN	O	O
findings	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
was	NN	O	O
stimulated	NN	O	O
18-fold	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
preformed	NN	O	O
,	NN	O	O
heterodimeric	NN	O	B-protein
NFKB2	NN	O	I-protein
(	NN	O	I-protein
p49	NN	O	I-protein
)	NN	O	I-protein
/RelA	NN	O	I-protein
(	NN	O	I-protein
p65	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	O
enhancer	NN	O	O
was	NN	O	O
also	NN	O	O
subject	NN	O	O
to	NN	O	O
regulation	NN	O	O
by	NN	O	O
recently	NN	O	O
cloned	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
(	NN	O	O
MAD-3	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Recombinant	NN	O	O
I	NN	O	O
kappa	NN	O	O
B-alpha	NN	O	O
(	NN	O	O
MAD-3	NN	O	B-protein
)	NN	O	O
inhibited	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
p49/p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p50/p65	NN	O	B-protein
but	NN	O	O
stimulated	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
NFKB2	NN	O	B-protein
(	NN	O	O
p49	NN	O	B-protein
)	NN	O	O
or	NN	O	O
NFKB1	NN	O	O
(	NN	O	O
p50	NN	O	O
)	NN	O	O
.	NN	O	O

Functional	NN	O	O
activation	NN	O	O
of	NN	O	O
an	NN	O	O
HIV	NN	O	O
reporter	NN	O	O
plasmid	NN	O	O
by	NN	O	O
p49/p65	NN	O	B-protein
in	NN	O	O
transiently	NN	O	B-cell_line
transfected	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
T-leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
also	NN	O	O
inhibited	NN	O	O
by	NN	O	O
coexpression	NN	O	O
of	NN	O	O
MAD-3	NN	O	B-protein
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

The	NN	O	O
c-rel	NN	O	B-protein
protooncogene	NN	O	I-protein
product	NN	O	I-protein
represses	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	B-protein
-mediated	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
long	NN	O	O
terminal	NN	O	O
repeat	NN	O	O
of	NN	O	O
type	NN	O	O
1	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
.	NN	O	O

The	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
type	NN	O	O
1	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
subunit	NN	O	I-protein
(	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
share	NN	O	O
functional	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
these	NN	O	O
inducible	NN	O	O
transcription	NN	O	O
units	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

These	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
are	NN	O	O
recognized	NN	O	O
by	NN	O	O
a	NN	O	O
structurally	NN	O	O
related	NN	O	O
family	NN	O	O
of	NN	O	O
interactive	NN	O	O
proteins	NN	O	O
that	NN	O	O
includes	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
c-rel	NN	O	B-DNA
protooncogene	NN	O	I-DNA
(	NN	O	O
c-Rel	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Recent	NN	O	O
biochemical	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
p50	NN	O	B-protein
form	NN	O	O
the	NN	O	O
prototypical	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
rapidly	NN	O	O
translocated	NN	O	O
from	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

This	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
complex	NN	O	O
potently	NN	O	O
stimulates	NN	O	O
kappa	NN	O	O
B-directed	NN	O	O
transcription	NN	O	O
from	NN	O	O
either	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
or	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
via	NN	O	O
the	NN	O	O
strong	NN	O	O
transactivation	NN	O	O
domain	NN	O	O
present	NN	O	O
in	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

We	NN	O	O
now	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
either	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
or	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
with	NN	O	O
delayed	NN	O	O
kinetics	NN	O	O
relative	NN	O	O
to	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
markedly	NN	O	O
represses	NN	O	O
p65	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
these	NN	O	O
transcription	NN	O	O
units	NN	O	O
.	NN	O	O

These	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
correlate	NN	O	O
with	NN	O	O
its	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
heterodimerize	NN	O	O
with	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
c-Rel	NN	O	B-protein
inhibition	NN	O	O
involves	NN	O	O
competition	NN	O	O
with	NN	O	O
p50/p65	NN	O	B-protein
for	NN	O	O
occupancy	NN	O	O
of	NN	O	O
the	NN	O	O
kappa	NN	O	O
B	NN	O	O
enhancer	NN	O	O
element	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
one	NN	O	O
function	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
is	NN	O	O
as	NN	O	O
a	NN	O	O
physiologic	NN	O	O
repressor	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
and	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
promoters	NN	O	O
,	NN	O	O
serving	NN	O	O
to	NN	O	O
efficiently	NN	O	O
counter	NN	O	O
the	NN	O	O
strong	NN	O	O
transcriptional	NN	O	O
activating	NN	O	O
effects	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Ras	NN	O	B-DNA
oncogene	NN	O	I-DNA
transformation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
and	NN	O	O
with	NN	O	O
a	NN	O	O
block	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
p21ras	NN	O	B-protein
small	NN	O	I-protein
GTP	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
participate	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
from	NN	O	O
cell	NN	O	O
surface	NN	O	O
receptors	NN	O	O
and	NN	O	O
affect	NN	O	O
neoplastic	NN	O	O
transformation	NN	O	O
and	NN	O	O
development	NN	O	O
in	NN	O	O
many	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
ras	NN	O	O
transformation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
T24	NN	O	B-DNA
Ha-ras	NN	O	I-DNA
oncogene	NN	O	I-DNA
in	NN	O	O
EBV-immortalized	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphoblasts	NN	O	I-cell_line
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
subunit	NN	O	I-protein
,	NN	O	O
with	NN	O	O
an	NN	O	O
impaired	NN	O	O
immunoglobulin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
altered	NN	O	O
adhesion	NN	O	O
properties	NN	O	O
and	NN	O	O
increased	NN	O	O
survival	NN	O	O
in	NN	O	O
serum-free	NN	O	O
medium	NN	O	O
.	NN	O	O

Since	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
subunit	NN	O	I-protein
is	NN	O	O
a	NN	O	O
hallmark	NN	O	O
of	NN	O	O
lymphocyte	NN	O	B-cell_type
activation	NN	O	O
,	NN	O	O
we	NN	O	O
suggest	NN	O	O
that	NN	O	O
p21ras	NN	O	B-protein
naturally	NN	O	O
triggers	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

The	NN	O	O
ras-transformed	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
displayed	NN	O	O
a	NN	O	O
fully	NN	O	O
functional	NN	O	O
IL-2r	NN	O	B-protein
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
c-fos	NN	O	B-DNA
induction	NN	O	O
following	NN	O	O
treatment	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
;	NN	O	O
nevertheless	NN	O	O
,	NN	O	O
they	NN	O	O
were	NN	O	O
not	NN	O	O
growth	NN	O	O
stimulated	NN	O	O
by	NN	O	O
this	NN	O	O
lymphokine	NN	O	B-protein
.	NN	O	O

The	NN	O	O
decreased	NN	O	O
expression	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
ras	NN	O	B-DNA
oncogene	NN	O	I-DNA
blocks	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
to	NN	O	O
plasma	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
possibly	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
lymphocyte-specific	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Somewhat	NN	O	O
unexpectedly	NN	O	O
,	NN	O	O
the	NN	O	O
constitutive	NN	O	O
p21ras	NN	O	B-protein
activity	NN	O	O
did	NN	O	O
not	NN	O	O
cause	NN	O	O
an	NN	O	O
increased	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
PEA1	NN	O	B-protein
(	NN	O	O
AP1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
PEA3	NN	O	B-protein
,	NN	O	O
Oct-2	NN	O	B-protein
or	NN	O	O
NF-kB	NN	O	B-protein

-DOCSTART-	O

Comparative	NN	O	O
analysis	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
(	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
)	NN	O	O
complex	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Nuclear	NN	O	O
factor	NN	O	O
of	NN	O	O
activated	NN	O	O
T	NN	O	O
cells	NN	O	O
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
to	NN	O	O
sequences	NN	O	O
in	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
largely	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
T	NN	O	O
cell-specific	NN	O	O
inducibility	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
(	NN	O	O
EMSA	NN	O	O
)	NN	O	O
showed	NN	O	O
that	NN	O	O
specific	NN	O	O
NFAT	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
could	NN	O	O
also	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
NFAT	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
however	NN	O	O
,	NN	O	O
was	NN	O	O
not	NN	O	O
functional	NN	O	O
,	NN	O	O
since	NN	O	O
it	NN	O	O
failed	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
from	NN	O	O
an	NN	O	O
NFAT-driven	NN	O	B-DNA
chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
construct	NN	O	I-DNA
.	NN	O	O

Competition	NN	O	O
with	NN	O	O
an	NN	O	O
AP-1	NN	O	B-DNA
motif	NN	O	I-DNA
or	NN	O	O
with	NN	O	O
anti-Jun	NN	O	B-protein
and	NN	O	O
anti-Fos	NN	O	B-protein
antibodies	NN	O	I-protein
abolished	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
motif	NN	O	I-DNA
in	NN	O	O
both	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
Jun	NN	O	O
and	NN	O	O
Fos	NN	O	O
are	NN	O	O
critical	NN	O	O
for	NN	O	O
NFAT	NN	O	B-protein
complex	NN	O	I-protein
formation	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Purified	NN	O	O
recombinant	NN	O	O
Jun	NN	O	B-protein
and	NN	O	I-protein
Fos	NN	O	I-protein
proteins	NN	O	I-protein
failed	NN	O	O
to	NN	O	O
bind	NN	O	O
directly	NN	O	O
to	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
motif	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
when	NN	O	O
combined	NN	O	O
with	NN	O	O
unstimulated	NN	O	O
B	NN	O	O
or	NN	O	O
T	NN	O	O
cell	NN	O	O
extracts	NN	O	O
,	NN	O	O
full-length	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
truncated	NN	O	O
,	NN	O	O
Jun/Fos	NN	O	O
heterodimers	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
form	NN	O	O
an	NN	O	O
NFAT	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
indicating	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
constitutively	NN	O	B-protein
expressed	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-protein
complex	NN	O	I-protein
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

An	NN	O	O
NFAT	NN	O	B-protein
oligonucleotide	NN	O	O
carrying	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
purine-rich	NN	O	I-DNA
part	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
sequence	NN	O	I-DNA
failed	NN	O	O
to	NN	O	O
form	NN	O	O
a	NN	O	O
complex	NN	O	O
and	NN	O	O
to	NN	O	O
compete	NN	O	O
with	NN	O	O
the	NN	O	O
wild	NN	O	O
type	NN	O	O
motif	NN	O	O
for	NN	O	O
NFAT	NN	O	B-protein
complex	NN	O	I-protein
formation	NN	O	O
in	NN	O	O
both	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
propose	NN	O	O
a	NN	O	O
model	NN	O	O
whereby	NN	O	O
a	NN	O	O
core	NN	O	O
NFAT	NN	O	B-protein
complex	NN	O	I-protein
consisting	NN	O	O
of	NN	O	O
Jun	NN	O	B-protein
,	NN	O	O
Fos	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
constitutive	NN	O	B-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
is	NN	O	O
formed	NN	O	O
in	NN	O	O
both	NN	O	O
T	NN	O	B-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
an	NN	O	O
additional	NN	O	O
factor	NN	O	O
and/or	NN	O	O
post-translational	NN	O	O
modification	NN	O	O
of	NN	O	O
a	NN	O	O
factor	NN	O	O
,	NN	O	O
missing	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
might	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
transactivation	NN	O	O
by	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Differential	NN	O	O
autoregulation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
T-	NN	O	I-cell_line
and	NN	O	I-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Regulation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	O
)	NN	O	O
expression	NN	O	O
by	NN	O	O
its	NN	O	O
cognate	NN	O	O
ligand	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
the	NN	O	O
glucocorticoid-sensitive	NN	O	B-cell_line
human	NN	O	I-cell_line
leukemic	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
6TG1.1	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
IM-9	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
decrease	NN	O	O
in	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
seen	NN	O	O
in	NN	O	O
IM-9	NN	O	B-cell_line
cells	NN	O	I-cell_line
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
1	NN	O	O
microM	NN	O	O
dexamethasone	NN	O	O
for	NN	O	O
16-18	NN	O	O
h	NN	O	O
,	NN	O	O
treatment	NN	O	O
of	NN	O	O
6TG1.1	NN	O	B-cell_line
cells	NN	O	I-cell_line
resulted	NN	O	O
in	NN	O	O
an	NN	O	O
8-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
and	NN	O	O
RNase	NN	O	B-protein
protection	NN	O	O
analysis	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
corresponding	NN	O	O
3-	NN	O	O
to	NN	O	O
4-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
GR	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Half-maximal	NN	O	O
induction	NN	O	O
of	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
in	NN	O	O
6TG1.1	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
observed	NN	O	O
between	NN	O	O
10-100	NN	O	O
nM	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
and	NN	O	O
inclusion	NN	O	O
of	NN	O	O
1	NN	O	O
microM	NN	O	O
RU	NN	O	O
38486	NN	O	O
completely	NN	O	O
blocked	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
100	NN	O	O
nM	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
positive	NN	O	O
autoregulation	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
6TG1.1	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
a	NN	O	O
receptor-mediated	NN	O	O
response	NN	O	O
.	NN	O	O

Positive	NN	O	O
autoregulation	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
also	NN	O	O
observed	NN	O	O
in	NN	O	O
glucocorticoid-resistant	NN	O	B-cell_line
CEM-C1	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
contain	NN	O	O
functional	NN	O	O
GR	NN	O	B-protein
,	NN	O	O
but	NN	O	O
whose	NN	O	O
growth	NN	O	O
is	NN	O	O
unaffected	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
positive	NN	O	O
autoregulation	NN	O	O
is	NN	O	O
neither	NN	O	O
a	NN	O	O
consequence	NN	O	O
nor	NN	O	O
the	NN	O	O
sole	NN	O	O
cause	NN	O	O
of	NN	O	O
growth	NN	O	O
arrest	NN	O	O
.	NN	O	O

The	NN	O	O
degree	NN	O	O
of	NN	O	O
negative	NN	O	O
autoregulation	NN	O	O
in	NN	O	O
IM-9	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
positive	NN	O	O
autoregulation	NN	O	O
in	NN	O	O
6TG1.1	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
with	NN	O	O
cycloheximide	NN	O	O
.	NN	O	O

Measurement	NN	O	O
of	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
turnover	NN	O	O
in	NN	O	O
6TG1.1	NN	O	B-cell_line
cells	NN	O	I-cell_line
treated	NN	O	O
with	NN	O	O
actinomycin-D	NN	O	O
revealed	NN	O	O
a	NN	O	O
half-life	NN	O	O
of	NN	O	O
2.5	NN	O	O
h	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
treatment	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
half-life	NN	O	O
was	NN	O	O
determined	NN	O	O
in	NN	O	O
IM-9	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
was	NN	O	O
also	NN	O	O
unaffected	NN	O	O
by	NN	O	O
steroid	NN	O	O
treatment	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
interpretation	NN	O	O
that	NN	O	O
glucocorticoid-mediated	NN	O	O
autoregulation	NN	O	O
of	NN	O	O
GR	NN	O	O
expression	NN	O	O
is	NN	O	O
a	NN	O	O
tissue-specific	NN	O	O
primary	NN	O	O
transcriptional	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
CD8	NN	O	B-protein
antigen	NN	O	I-protein
and	NN	O	O
suppressor	NN	O	O
activity	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
a	NN	O	O
CEM	NN	O	B-cell_line
human	NN	O	I-cell_line
leukemic	NN	O	I-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
relationship	NN	O	O
between	NN	O	O
glucocorticoid	NN	O	O
effect	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
antigens	NN	O	I-protein
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
two	NN	O	O
models	NN	O	O
of	NN	O	O
leukemic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
CEM	NN	O	B-cell_line
C7	NN	O	I-cell_line
denoted	NN	O	O
(	NN	O	O
r+	NN	O	O
,	NN	O	O
ly+	NN	O	O
)	NN	O	O
and	NN	O	O
CEM	NN	O	B-cell_line
C1	NN	O	I-cell_line
(	NN	O	O
r+	NN	O	O
,	NN	O	O
ly-	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
reactivity	NN	O	O
of	NN	O	O
murine	NN	O	B-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
,	NN	O	O
anti-CD4-FITC	NN	O	B-protein
,	NN	O	O
anti-CD8-FITC	NN	O	B-protein
,	NN	O	O
anti-CD2-FITC	NN	O	B-protein
and	NN	O	O
anti-calla-FITC	NN	O	B-protein
,	NN	O	O
were	NN	O	O
analyzed	NN	O	O
using	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
.	NN	O	O

The	NN	O	O
suppressor	NN	O	O
function	NN	O	O
was	NN	O	O
determined	NN	O	O
using	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
into	NN	O	O
phytohemagglutinin-activated	NN	O	B-cell_line
peripheral	NN	O	I-cell_line
blood	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Dexamethasone	NN	O	O
treatment	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
CEM	NN	O	I-cell_line
C7	NN	O	I-cell_line
caused	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
surface	NN	O	B-protein
antigen	NN	O	I-protein
CD8	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
present	NN	O	O
on	NN	O	O
suppressor	NN	O	O
and	NN	O	O
cytotoxic	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
.	NN	O	O

By	NN	O	O
comparison	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
modification	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
antigen	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
high	NN	O	O
levels	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

After	NN	O	O
two	NN	O	O
days	NN	O	O
of	NN	O	O
treatment	NN	O	O
with	NN	O	O
5	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
-8	NN	O	O
)	NN	O	O
M	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
CEM	NN	O	B-cell_line
C7	NN	O	I-cell_line
cells	NN	O	I-cell_line
showed	NN	O	O
a	NN	O	O
two-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
suppressor	NN	O	O
activity	NN	O	O
compared	NN	O	O
to	NN	O	O
untreated	NN	O	O
cells	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
regulation	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
CD8	NN	O	B-protein
or	NN	O	O
CD4	NN	O	B-protein
antigens	NN	O	I-protein
in	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_line
clone	NN	O	I-cell_line
CEM	NN	O	I-cell_line
C1	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
no	NN	O	O
modification	NN	O	O
of	NN	O	O
the	NN	O	O
suppressor	NN	O	O
function	NN	O	O
in	NN	O	O
CEM	NN	O	B-cell_line
C1	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
human	NN	O	O
leukemic	NN	O	O
cells	NN	O	O
studied	NN	O	O
here	NN	O	O
,	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
CD8	NN	O	B-protein
antigen	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
a	NN	O	O
CD4+	NN	O	B-cell_line
cells	NN	O	I-cell_line
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
cell	NN	O	O
lysis	NN	O	O
in	NN	O	O
a	NN	O	O
glucocorticoid	NN	O	B-cell_line
receptor	NN	O	I-cell_line
positive	NN	O	I-cell_line
cell	NN	O	I-cell_line
population	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Adenovirus	NN	O	O
E1A	NN	O	O
inhibits	NN	O	O
IFN	NN	O	B-protein
-induced	NN	O	O
resistance	NN	O	O
to	NN	O	O
cytolysis	NN	O	O
by	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Infection	NN	O	O
of	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
cytopathic	NN	O	O
viruses	NN	O	O
inhibits	NN	O	O
IFN	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
cytolytic	NN	O	O
resistance	NN	O	O
to	NN	O	O
NK	NN	O	O
cell-mediated	NN	O	O
cytolysis	NN	O	O
[	NN	O	O
IFN	NN	O	B-protein
-mediated	NN	O	O
cytoprotection	NN	O	O
(	NN	O	O
IFN	NN	O	B-protein
-MCP	NN	O	O
)	NN	O	O
]	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
thought	NN	O	O
that	NN	O	O
the	NN	O	O
virally	NN	O	O
induced	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IFN	NN	O	B-protein
-MCP	NN	O	O
is	NN	O	O
secondary	NN	O	O
to	NN	O	O
the	NN	O	O
shutdown	NN	O	O
of	NN	O	O
cellular	NN	O	O
macromolecular	NN	O	O
synthesis	NN	O	O
that	NN	O	O
accompanies	NN	O	O
cytopathic	NN	O	O
virus	NN	O	O
infections	NN	O	O
.	NN	O	O

Group	NN	O	O
C	NN	O	O
,	NN	O	O
adenovirus	NN	O	O
serotype	NN	O	O
5	NN	O	O
(	NN	O	O
Ad5	NN	O	O
)	NN	O	O
infection	NN	O	O
inhibits	NN	O	O
both	NN	O	O
IFN	NN	O	B-protein
-MCP	NN	O	O
and	NN	O	O
cellular	NN	O	B-protein
protein	NN	O	I-protein
synthesis	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
determined	NN	O	O
if	NN	O	O
the	NN	O	O
Ad5-induced	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IFN	NN	O	B-protein
-MCP	NN	O	O
was	NN	O	O
independent	NN	O	O
of	NN	O	O
adenovirus	NN	O	O
(	NN	O	O
Ad	NN	O	O
)	NN	O	O
infection	NN	O	O
and	NN	O	O
secondary	NN	O	O
only	NN	O	O
to	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Ad	NN	O	B-DNA
early	NN	O	I-DNA
region	NN	O	I-DNA
1A	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
E1A	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
4-h	NN	O	O
NK	NN	O	O
cytolysis	NN	O	O
assays	NN	O	O
were	NN	O	O
performed	NN	O	O
on	NN	O	O
IFN-gamma-treated	NN	O	B-cell_line
human	NN	O	I-cell_line
cells	NN	O	I-cell_line
infected	NN	O	O
with	NN	O	O
an	NN	O	O
Ad5	NN	O	O
E1A	NN	O	B-DNA
deletion	NN	O	O
mutant	NN	O	O
,	NN	O	O
dl343	NN	O	O
,	NN	O	O
or	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
Ad5	NN	O	B-DNA
E1A	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

IFN	NN	O	B-protein
-MCP	NN	O	O
was	NN	O	O
not	NN	O	O
inhibited	NN	O	O
by	NN	O	O
infection	NN	O	O
with	NN	O	O
dl343	NN	O	O
,	NN	O	O
despite	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
large	NN	O	O
amounts	NN	O	O
of	NN	O	O
both	NN	O	O
early	NN	O	O
(	NN	O	O
E1B	NN	O	B-protein
,	NN	O	O
p55	NN	O	B-protein
)	NN	O	O
and	NN	O	O
late	NN	O	O
(	NN	O	O
hexon	NN	O	B-protein
)	NN	O	O
Ad	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
E1A-negative	NN	O	B-cell_line
,	NN	O	I-cell_line
parental	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
IFN	NN	O	B-protein
-MCP	NN	O	O
was	NN	O	O
blocked	NN	O	O
in	NN	O	O
Ad5	NN	O	O
E1A-transfected	NN	O	B-cell_line
epithelial	NN	O	I-cell_line
and	NN	O	O
fibroblastic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Genetic	NN	O	O
mapping	NN	O	O
studies	NN	O	O
within	NN	O	O
the	NN	O	O
E1A	NN	O	B-DNA
gene	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
only	NN	O	O
the	NN	O	O
first	NN	O	O
exon	NN	O	O
of	NN	O	O
E1A	NN	O	B-DNA
was	NN	O	O
sufficient	NN	O	O
to	NN	O	O
inhibit	NN	O	O
IFN	NN	O	B-protein
-MCP	NN	O	O
.	NN	O	O

DNA	NN	O	O
sequence	NN	O	O
homology	NN	O	O
of	NN	O	O
E1A	NN	O	B-DNA
genes	NN	O	O
between	NN	O	O
different	NN	O	O
Ad	NN	O	O
groups	NN	O	O
(	NN	O	O
group	NN	O	O
A	NN	O	O
,	NN	O	O
Ad12	NN	O	O
;	NN	O	O
group	NN	O	O
C	NN	O	O
,	NN	O	O
Ad5	NN	O	O
)	NN	O	O
is	NN	O	O
limited	NN	O	O
almost	NN	O	O
entirely	NN	O	O
to	NN	O	O
three	NN	O	O
conserved	NN	O	O
regions	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
first	NN	O	O
exon	NN	O	O
of	NN	O	O
E1A	NN	O	B-DNA
.	NN	O	O

Because	NN	O	O
IFN	NN	O	B-protein
-MCP	NN	O	O
was	NN	O	O
also	NN	O	O
blocked	NN	O	O
in	NN	O	O
Ad12	NN	O	B-cell_line
E1A-transfected	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
expression	NN	O	O
of	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
of	NN	O	O
the	NN	O	O
E1A-conserved	NN	O	B-DNA
regions	NN	O	I-DNA
may	NN	O	O
be	NN	O	O
necessary	NN	O	O
to	NN	O	O
inhibit	NN	O	O
IFN	NN	O	B-protein
-MCP	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
E1A	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
inhibited	NN	O	O
IFN	NN	O	B-protein
-MCP	NN	O	O
independently	NN	O	O
of	NN	O	O
virus	NN	O	O
infection	NN	O	O
.	NN	O	O

E1A	NN	O	B-DNA
's	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IFN	NN	O	B-protein
-MCP	NN	O	O
has	NN	O	O
the	NN	O	O
net	NN	O	O
effect	NN	O	O
of	NN	O	O
promoting	NN	O	O
the	NN	O	O
selective	NN	O	O
NK	NN	O	O
cell-mediated	NN	O	O
clearance	NN	O	O
of	NN	O	O
Ad-infected	NN	O	O
or	NN	O	O
Ad-transformed	NN	O	B-cell_line
human	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
protein	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
family	NN	O	I-protein
is	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
.	NN	O	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
(	NN	O	O
NF-AT	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
response	NN	O	O
element	NN	O	O
for	NN	O	O
NF-AT	NN	O	B-protein
is	NN	O	O
a	NN	O	O
sequence	NN	O	O
localized	NN	O	O
between	NN	O	O
-285/-254	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
composition	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
protein	NN	O	O
is	NN	O	O
still	NN	O	O
not	NN	O	O
fully	NN	O	O
elucidated	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
normal	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
an	NN	O	O
AP-1	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
protein	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

This	NN	O	O
was	NN	O	O
evidenced	NN	O	O
by	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
to	NN	O	O
compete	NN	O	O
with	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
site	NN	O	O
for	NN	O	O
binding	NN	O	O
to	NN	O	O
NF-AT	NN	O	B-protein
and	NN	O	O
by	NN	O	O
the	NN	O	O
capacity	NN	O	O
of	NN	O	O
immobilized	NN	O	B-protein
anti-Jun	NN	O	I-protein
and	NN	O	O
anti-Fos	NN	O	B-protein
antibodies	NN	O	O
to	NN	O	O
deplete	NN	O	O
NF-AT	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
from	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
detectable	NN	O	O
binding	NN	O	O
of	NN	O	O
in	NN	O	O
vitro	NN	O	O
translated	NN	O	O
Jun/Fos	NN	O	B-protein
heterodimer	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
to	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
sequence	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-DNA
sequence	NN	O	I-DNA
was	NN	O	O
unable	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
Jun/Fos	NN	O	B-protein
to	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
AP-1	NN	O	B-protein
protein	NN	O	O
in	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
protein	NN	O	O
complex	NN	O	O
may	NN	O	O
regulate	NN	O	O
NF-AT	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
through	NN	O	O
protein-protein	NN	O	O
interaction	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interaction	NN	O	O
between	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-and	NN	O	O
serum	NN	O	B-protein
response	NN	O	I-protein
factor	NN	O	I-protein
-binding	NN	O	O
elements	NN	O	O
activates	NN	O	O
an	NN	O	O
interleukin-2	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha-chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
specifically	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
a	NN	O	O
short	NN	O	O
enhancer	NN	O	O
element	NN	O	O
containing	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha-chain	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
is	NN	O	O
preferentially	NN	O	O
activated	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
enhancer	NN	O	O
binds	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
poorly	NN	O	O
and	NN	O	O
is	NN	O	O
only	NN	O	O
weakly	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
alone	NN	O	O
.	NN	O	O

Serum	NN	O	B-protein
response	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
SRF	NN	O	B-protein
)	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
site	NN	O	O
adjacent	NN	O	O
to	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
both	NN	O	O
sites	NN	O	O
together	NN	O	O
have	NN	O	O
strong	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
specifically	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
SRF	NN	O	B-protein
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
consistently	NN	O	O
higher	NN	O	O
than	NN	O	O
in	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
SRF	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
causes	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
enhancer	NN	O	I-DNA
to	NN	O	O
function	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
it	NN	O	O
does	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
SRF	NN	O	B-protein
binding	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
functionally	NN	O	O
important	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mutual	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
its	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
NK-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complex	NN	O	I-protein
is	NN	O	O
sequestered	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
by	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
protein	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
(	NN	O	O
MAD-3	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Various	NN	O	O
cellular	NN	O	O
stimuli	NN	O	O
relieve	NN	O	O
this	NN	O	O
inhibition	NN	O	O
by	NN	O	O
mechanisms	NN	O	O
largely	NN	O	O
unknown	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
nuclear	NN	O	O
localization	NN	O	O
and	NN	O	O
transactivation	NN	O	O
of	NN	O	O
its	NN	O	O
target	NN	O	O
genes	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
demonstrated	NN	O	O
here	NN	O	O
with	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
that	NN	O	O
different	NN	O	O
stimuli	NN	O	O
,	NN	O	O
including	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
,	NN	O	O
cause	NN	O	O
rapid	NN	O	O
degradation	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
,	NN	O	O
with	NN	O	O
concomitant	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
followed	NN	O	O
by	NN	O	O
a	NN	O	O
dramatic	NN	O	O
increase	NN	O	O
in	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B-alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Transfection	NN	O	O
studies	NN	O	O
reveal	NN	O	O
that	NN	O	O
the	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B-alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
the	NN	O	O
encoded	NN	O	O
protein	NN	O	O
are	NN	O	O
potently	NN	O	O
induced	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
by	NN	O	O
homodimers	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
.	NN	O	O

We	NN	O	O
propose	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
mutually	NN	O	O
regulate	NN	O	O
each	NN	O	O
other	NN	O	O
in	NN	O	O
a	NN	O	O
cycle	NN	O	O
:	NN	O	O
saturating	NN	O	O
amounts	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
protein	NN	O	O
are	NN	O	O
destroyed	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
,	NN	O	O
allowing	NN	O	O
rapid	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Subsequently	NN	O	O
,	NN	O	O
I	NN	O	B-RNA
kappa	NN	O	I-RNA
B-alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
levels	NN	O	O
are	NN	O	O
quickly	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

This	NN	O	O
resurgence	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
protein	NN	O	O
acts	NN	O	O
to	NN	O	O
restore	NN	O	O
an	NN	O	O
equilibrium	NN	O	O
in	NN	O	O
which	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
again	NN	O	O
inhibited	NN	O	O
.	NN	O	O

-DOCSTART-	O

Replication	NN	O	O
of	NN	O	O
type	NN	O	O
1	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
viruses	NN	O	O
containing	NN	O	O
linker	NN	O	O
substitution	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
-201	NN	O	B-DNA
to	NN	O	I-DNA
-130	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
previous	NN	O	O
transfection	NN	O	O
analyses	NN	O	O
using	NN	O	O
the	NN	O	O
chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
system	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
that	NN	O	O
linker	NN	O	O
substitution	NN	O	O
(	NN	O	O
LS	NN	O	O
)	NN	O	O
mutations	NN	O	O
between	NN	O	O
-201	NN	O	B-DNA
and	NN	O	I-DNA
-130	NN	O	I-DNA
(	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	O
start	NN	O	O
site	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
caused	NN	O	O
moderate	NN	O	O
decreases	NN	O	O
in	NN	O	O
LTR	NN	O	B-DNA
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
T-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
(	NN	O	O
S.L.Zeichner	NN	O	O
,	NN	O	O
J.Y.H.	NN	O	O
Kim	NN	O	O
,	NN	O	O
and	NN	O	O
J.C.Alwine	NN	O	O
,	NN	O	O
J.Virol.65	NN	O	O
:	NN	O	O
2436-2444	NN	O	O
,	NN	O	O
1991	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
confirm	NN	O	O
the	NN	O	O
significance	NN	O	O
of	NN	O	O
this	NN	O	O
region	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
viral	NN	O	O
replication	NN	O	O
,	NN	O	O
we	NN	O	O
constructed	NN	O	O
several	NN	O	O
of	NN	O	O
these	NN	O	O
LS	NN	O	O
mutations	NN	O	O
(	NN	O	O
-201	NN	O	B-DNA
to	NN	O	I-DNA
-	NN	O	I-DNA
184	NN	O	I-DNA
,	NN	O	O
-183	NN	O	B-DNA
to	NN	O	I-DNA
-166	NN	O	I-DNA
,	NN	O	O
-165	NN	O	B-DNA
to	NN	O	I-DNA
-148	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
-148	NN	O	B-DNA
to	NN	O	I-DNA
-130	NN	O	I-DNA
)	NN	O	O
in	NN	O	O
proviruses	NN	O	O
and	NN	O	O
prepared	NN	O	O
viral	NN	O	O
stocks	NN	O	O
by	NN	O	O
cocultivation	NN	O	O
of	NN	O	O
transfected	NN	O	B-cell_line
RD	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
CEMx174	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
two	NN	O	O
mutations	NN	O	O
between	NN	O	O
-93	NN	O	B-DNA
and	NN	O	I-DNA
-76	NN	O	I-DNA
and	NN	O	O
between	NN	O	O
-75	NN	O	B-DNA
and	NN	O	I-DNA
-58	NN	O	I-DNA
were	NN	O	O
utilized	NN	O	O
,	NN	O	O
since	NN	O	O
they	NN	O	O
affect	NN	O	O
the	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
(	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
)	NN	O	I-DNA
-	NN	O	I-DNA
and	NN	O	I-DNA
Sp1-binding	NN	O	I-DNA
sites	NN	O	I-DNA
and	NN	O	O
were	NN	O	O
expected	NN	O	O
to	NN	O	O
diminish	NN	O	O
viral	NN	O	O
replication	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
while	NN	O	O
transfection	NN	O	O
analyses	NN	O	O
offer	NN	O	O
an	NN	O	O
adequate	NN	O	O
approximation	NN	O	O
of	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
LS	NN	O	O
mutations	NN	O	O
,	NN	O	O
the	NN	O	O
analysis	NN	O	O
of	NN	O	O
viral	NN	O	O
replication	NN	O	O
using	NN	O	O
a	NN	O	O
mutant	NN	O	O
viral	NN	O	O
stock	NN	O	O
presents	NN	O	O
a	NN	O	O
more	NN	O	O
accurate	NN	O	O
picture	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
sometimes	NN	O	O
at	NN	O	O
variance	NN	O	O
with	NN	O	O
the	NN	O	O
transfection	NN	O	O
results	NN	O	O
.	NN	O	O

Three	NN	O	O
mutants	NN	O	O
(	NN	O	O
-201/-184	NN	O	O
NXS	NN	O	O
,	NN	O	O
-165/-148	NN	O	O
NXS	NN	O	O
,	NN	O	O
and	NN	O	O
-147/-130	NN	O	O
NXS	NN	O	O
)	NN	O	O
had	NN	O	O
effects	NN	O	O
on	NN	O	O
viral	NN	O	O
replication	NN	O	O
that	NN	O	O
were	NN	O	O
much	NN	O	O
more	NN	O	O
severe	NN	O	O
than	NN	O	O
the	NN	O	O
effects	NN	O	O
predicted	NN	O	O
from	NN	O	O
their	NN	O	O
performance	NN	O	O
in	NN	O	O
transfection	NN	O	O
analyses	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
two	NN	O	O
LS	NN	O	O
mutations	NN	O	O
(	NN	O	O
-201/-184	NN	O	O
NXS	NN	O	O
and	NN	O	O
-183/-166	NN	O	O
NXS	NN	O	O
)	NN	O	O
were	NN	O	O
not	NN	O	O
predicted	NN	O	O
by	NN	O	O
their	NN	O	O
effects	NN	O	O
in	NN	O	O
transfection	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
permissiveness	NN	O	O
to	NN	O	O
replication	NN	O	O
of	NN	O	O
some	NN	O	O
mutant	NN	O	O
viruses	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
cell	NN	O	O
types	NN	O	O
tested	NN	O	O
,	NN	O	O
the	NN	O	O
LS	NN	O	O
mutations	NN	O	O
indicated	NN	O	O
an	NN	O	O
apparent	NN	O	O
requirement	NN	O	O
not	NN	O	O
only	NN	O	O
for	NN	O	O
the	NN	O	O
intact	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
SP1-binding	NN	O	B-DNA
sites	NN	O	I-DNA
but	NN	O	O
also	NN	O	O
for	NN	O	O
several	NN	O	O
regions	NN	O	O
between	NN	O	O
-201	NN	O	B-DNA
and	NN	O	I-DNA
-130	NN	O	I-DNA
not	NN	O	O
previously	NN	O	O
associated	NN	O	O
with	NN	O	O
viral	NN	O	O
infectivity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
and	NN	O	O
erythroid	NN	O	O
development	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
we	NN	O	O
derived	NN	O	O
an	NN	O	O
experimental	NN	O	O
system	NN	O	O
that	NN	O	O
allows	NN	O	O
us	NN	O	O
to	NN	O	O
dissect	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
in	NN	O	O
red	NN	O	B-cell_type
cell	NN	O	I-cell_type
development	NN	O	O
at	NN	O	O
a	NN	O	O
genetic	NN	O	O
level	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
established	NN	O	O
the	NN	O	O
essential	NN	O	O
nature	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
during	NN	O	O
both	NN	O	O
primitive	NN	O	O
and	NN	O	O
definitive	NN	O	O
erythropoiesis	NN	O	O
.	NN	O	O

By	NN	O	O
ablating	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	O
GATA-1	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
are	NN	O	O
in	NN	O	O
a	NN	O	O
position	NN	O	O
to	NN	O	O
introduce	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
constructs	NN	O	O
that	NN	O	O
harbor	NN	O	O
subtle	NN	O	O
modifications	NN	O	O
in	NN	O	O
flanking	NN	O	O
or	NN	O	O
protein-coding	NN	O	B-DNA
sequences	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
can	NN	O	O
now	NN	O	O
study	NN	O	O
regulatory	NN	O	O
regions	NN	O	O
and	NN	O	O
functional	NN	O	O
domains	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
a	NN	O	O
true	NN	O	O
erythroid	NN	O	O
environment	NN	O	O
,	NN	O	O
experiments	NN	O	O
that	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
possible	NN	O	O
heretofore	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
assay	NN	O	O
involves	NN	O	O
the	NN	O	O
dramatic	NN	O	O
loss	NN	O	O
of	NN	O	O
red	NN	O	O
cell	NN	O	O
production	NN	O	O
,	NN	O	O
it	NN	O	O
should	NN	O	O
be	NN	O	O
possible	NN	O	O
to	NN	O	O
define	NN	O	O
important	NN	O	O
regulatory	NN	O	O
domains	NN	O	O
that	NN	O	O
can	NN	O	O
then	NN	O	O
be	NN	O	O
assayed	NN	O	O
using	NN	O	O
less	NN	O	O
stringent	NN	O	O
systems	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
cell-free	NN	O	O
extracts	NN	O	O
for	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
ideal	NN	O	O
situation	NN	O	O
would	NN	O	O
be	NN	O	O
analyses	NN	O	O
conducted	NN	O	O
in	NN	O	O
GATA-1	NN	O	B-cell_line
-erythroid	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
these	NN	O	O
cells	NN	O	O
have	NN	O	O
been	NN	O	O
impossible	NN	O	O
to	NN	O	O
generate	NN	O	O
given	NN	O	O
the	NN	O	O
requirement	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
for	NN	O	O
Epo	NN	O	B-protein
receptor	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
red	NN	O	B-cell_type
cell	NN	O	I-cell_type
viability	NN	O	O
(	NN	O	O
C.	NN	O	O
Simon	NN	O	O
and	NN	O	O
S.	NN	O	O
Orkin	NN	O	O
,	NN	O	O
unpublished	NN	O	O
observations	NN	O	O
)	NN	O	O
.	NN	O	O

It	NN	O	O
may	NN	O	O
be	NN	O	O
possible	NN	O	O
to	NN	O	O
produce	NN	O	O
such	NN	O	O
cells	NN	O	O
by	NN	O	O
first	NN	O	O
expressing	NN	O	O
the	NN	O	O
Epo	NN	O	B-protein
receptor	NN	O	I-protein
under	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
a	NN	O	O
constitutive	NN	O	O
promoter	NN	O	O
and	NN	O	O
then	NN	O	O
targeting	NN	O	O
the	NN	O	O
GATA-1	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

If	NN	O	O
GATA-1	NN	O	B-cell_line
-red	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
available	NN	O	O
,	NN	O	O
the	NN	O	O
analyses	NN	O	O
would	NN	O	O
involve	NN	O	O
the	NN	O	O
actual	NN	O	O
transcription	NN	O	O
of	NN	O	O
or	NN	O	O
chromatin	NN	O	O
structure	NN	O	O
surrounding	NN	O	O
the	NN	O	O
globin	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Structure-function	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
GATA-1	NN	O	B-protein
protein	NN	O	O
could	NN	O	O
be	NN	O	O
greatly	NN	O	O
simplified	NN	O	O
and	NN	O	O
a	NN	O	O
larger	NN	O	O
number	NN	O	O
of	NN	O	O
mutants	NN	O	O
studied	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
ES	NN	O	O
cell	NN	O	O
system	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
as	NN	O	O
an	NN	O	O
alternative	NN	O	O
until	NN	O	O
targeted	NN	O	B-cell_line
erythroleukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
become	NN	O	O
available	NN	O	O
.	NN	O	O

Other	NN	O	O
applications	NN	O	O
involve	NN	O	O
the	NN	O	O
introduction	NN	O	O
of	NN	O	O
other	NN	O	O
GATA-binding	NN	O	B-protein
protein	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
they	NN	O	O
rescue	NN	O	O
the	NN	O	O
mutation	NN	O	O
.	NN	O	O

If	NN	O	O
they	NN	O	O
can	NN	O	O
not	NN	O	O
,	NN	O	O
chimeric	NN	O	B-protein
proteins	NN	O	I-protein
can	NN	O	O
be	NN	O	O
tested	NN	O	O
to	NN	O	O
identify	NN	O	O
which	NN	O	O
amino	NN	O	O
acids	NN	O	O
distinguish	NN	O	O
the	NN	O	O
different	NN	O	O
family	NN	O	O
members	NN	O	O
.	NN	O	O

We	NN	O	O
feel	NN	O	O
that	NN	O	O
these	NN	O	O
experiments	NN	O	O
are	NN	O	O
vital	NN	O	O
to	NN	O	O
understanding	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
during	NN	O	O
erythroid	NN	O	O
ontogeny	NN	O	O
.	NN	O	O

How	NN	O	O
does	NN	O	O
GATA-1	NN	O	B-protein
regulate	NN	O	O
red	NN	O	B-DNA
cell	NN	O	I-DNA
genes	NN	O	I-DNA
like	NN	O	O
globin	NN	O	B-DNA
or	NN	O	O
the	NN	O	O
Epo	NN	O	B-protein
receptor	NN	O	I-protein
?	NN	O	O
Once	NN	O	O
we	NN	O	O
identify	NN	O	O
the	NN	O	O
functional	NN	O	O
domains	NN	O	O
of	NN	O	O
the	NN	O	O
GATA-binding	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
we	NN	O	O
hope	NN	O	O
to	NN	O	O
learn	NN	O	O
what	NN	O	O
proteins	NN	O	O
GATA-1	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
in	NN	O	O
the	NN	O	O
basic	NN	O	O
transcription	NN	O	O
machinery	NN	O	O
or	NN	O	O
in	NN	O	O
chromatin	NN	O	O
.	NN	O	O

Is	NN	O	O
GATA-1	NN	O	B-protein
necessary	NN	O	O
for	NN	O	O
globin	NN	O	O
gene	NN	O	O
switching	NN	O	O
?	NN	O	O
GATA-1	NN	O	B-protein
may	NN	O	O
be	NN	O	O
modified	NN	O	O
differently	NN	O	O
during	NN	O	O
development	NN	O	O
so	NN	O	O
that	NN	O	O
the	NN	O	O
locus	NN	O	O
control	NN	O	O
region	NN	O	O
can	NN	O	O
interact	NN	O	O
with	NN	O	O
different	NN	O	O
globin	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
may	NN	O	O
find	NN	O	O
that	NN	O	O
one	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
embryonic	NN	O	O
expression	NN	O	O
and	NN	O	O
another	NN	O	O
for	NN	O	O
adult	NN	O	O
globin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
human	NN	O	B-DNA
putative	NN	O	I-DNA
lymphocyte	NN	O	I-DNA
G0/G1	NN	O	I-DNA
switch	NN	O	I-DNA
gene	NN	O	I-DNA
homologous	NN	O	O
to	NN	O	O
a	NN	O	O
rodent	NN	O	B-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
a	NN	O	O
zinc-binding	NN	O	B-protein
potential	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

G0S24	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
genes	NN	O	O
(	NN	O	O
putative	NN	O	B-DNA
G0/G1	NN	O	I-DNA
switch	NN	O	I-DNA
regulatory	NN	O	I-DNA
genes	NN	O	I-DNA
)	NN	O	O
that	NN	O	O
are	NN	O	O
expressed	NN	O	O
transiently	NN	O	O
within	NN	O	O
1-2	NN	O	O
hr	NN	O	O
of	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
lectin	NN	O	B-protein
or	NN	O	O
cycloheximide	NN	O	O
to	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
a	NN	O	O
full-length	NN	O	B-DNA
cDNA	NN	O	I-DNA
sequence	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
corresponding	NN	O	O
genomic	NN	O	O
sequence	NN	O	O
reveals	NN	O	O
an	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
of	NN	O	O
326	NN	O	O
amino	NN	O	O
acids	NN	O	O
,	NN	O	O
distributed	NN	O	O
across	NN	O	O
two	NN	O	O
exons	NN	O	B-DNA
.	NN	O	O

Potential	NN	O	O
phosphorylation	NN	O	B-protein
sites	NN	O	I-protein
include	NN	O	O
the	NN	O	O
sequence	NN	O	O
PSPTSPT	NN	O	B-protein
,	NN	O	O
which	NN	O	O
resembles	NN	O	O
an	NN	O	O
RNA	NN	O	B-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
repeat	NN	O	I-protein
reported	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	B-protein
cycle	NN	O	I-protein
control	NN	O	I-protein
kinase	NN	O	I-protein
cdc2	NN	O	I-protein
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
the	NN	O	O
derived	NN	O	O
protein	NN	O	O
sequence	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
rodent	NN	O	O
homologs	NN	O	O
allows	NN	O	O
classification	NN	O	O
into	NN	O	O
three	NN	O	O
groups	NN	O	O
.	NN	O	O

Group	NN	O	O
1	NN	O	O
contains	NN	O	O
G0S24	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
rat	NN	O	B-DNA
and	NN	O	I-DNA
mouse	NN	O	I-DNA
TIS11	NN	O	I-DNA
genes	NN	O	I-DNA
(	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
TTP	NN	O	B-DNA
,	NN	O	O
Nup475	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
Zfp36	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Members	NN	O	O
of	NN	O	O
this	NN	O	O
group	NN	O	O
have	NN	O	O
three	NN	O	B-protein
tetraproline	NN	O	I-protein
repeats	NN	O	I-protein
.	NN	O	O

Groups	NN	O	O
1	NN	O	O
and	NN	O	O
2	NN	O	O
have	NN	O	O
a	NN	O	O
serine-rich	NN	O	B-protein
region	NN	O	I-protein
and	NN	O	O
an	NN	O	O
``	NN	O	O
arginine	NN	O	B-protein
element	NN	O	I-protein
``	NN	O	O
(	NN	O	O
RRLPIF	NN	O	B-protein
)	NN	O	O
at	NN	O	O
the	NN	O	O
carboxyl	NN	O	B-protein
terminus	NN	O	I-protein
.	NN	O	O

All	NN	O	O
groups	NN	O	O
contain	NN	O	O
cysteine-	NN	O	B-protein
and	NN	O	I-protein
histidine-rich	NN	O	I-protein
putative	NN	O	I-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
domains	NN	O	I-protein
and	NN	O	O
a	NN	O	O
serine-phenylalanine	NN	O	B-protein
``	NN	O	I-protein
SFS	NN	O	I-protein
''	NN	O	I-protein
domain	NN	O	I-protein
similar	NN	O	O
to	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
large	NN	O	O
subunit	NN	O	O
of	NN	O	O
eukaryotic	NN	O	B-protein
RNA	NN	O	I-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
group	NN	O	O
1	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
mouse	NN	O	I-DNA
genomic	NN	O	I-DNA
sequences	NN	O	I-DNA
shows	NN	O	O
high	NN	O	O
conservation	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flank	NN	O	I-DNA
and	NN	O	O
exons	NN	O	B-DNA
.	NN	O	O

A	NN	O	O
CpG	NN	O	O
island	NN	O	O
suggests	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
germ	NN	O	O
line	NN	O	O
.	NN	O	O

G0S24	NN	O	B-DNA
has	NN	O	O
potential	NN	O	O
sites	NN	O	O
for	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flank	NN	O	I-DNA
and	NN	O	O
intron	NN	O	B-DNA
;	NN	O	O
these	NN	O	O
include	NN	O	O
a	NN	O	O
serum	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Protein	NN	O	O
and	NN	O	O
genomic	NN	O	O
sequences	NN	O	O
show	NN	O	O
similarities	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
proteins	NN	O	O
involved	NN	O	O
in	NN	O	O
transcription	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
G0S24	NN	O	B-protein
product	NN	O	I-protein
has	NN	O	O
a	NN	O	O
similar	NN	O	O
role	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
lymphoid-specific	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
mu	NN	O	I-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
enhancer	NN	O	I-DNA
by	NN	O	O
ETS-domain	NN	O	O
proteins	NN	O	O
.	NN	O	O

The	NN	O	O
enhancer	NN	O	B-DNA
for	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
mu	NN	O	I-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
IgH	NN	O	B-protein
)	NN	O	O
activates	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
gene	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
pre-B	NN	O	O
cell	NN	O	O
stage	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
differentiation	NN	O	O
.	NN	O	O

A	NN	O	O
lymphoid-specific	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
microB	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
enhancer	NN	O	B-DNA
function	NN	O	O
in	NN	O	O
pre-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
microB	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
PU.1/Spi-1	NN	O	B-DNA
proto-oncogene	NN	O	I-DNA
.	NN	O	O

Another	NN	O	O
sequence	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
microA	NN	O	B-DNA
,	NN	O	O
was	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
mu	NN	O	B-DNA
enhancer	NN	O	I-DNA
that	NN	O	O
binds	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
ets-1	NN	O	B-DNA
proto-oncogene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
microA	NN	O	B-DNA
motif	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
microB-dependent	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
,	NN	O	O
which	NN	O	O
suggests	NN	O	O
that	NN	O	O
a	NN	O	O
minimal	NN	O	B-DNA
B	NN	O	I-DNA
cell-specific	NN	O	I-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
composed	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
PU.1	NN	O	B-DNA
and	NN	O	I-DNA
Ets-1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

Co-expression	NN	O	O
of	NN	O	O
both	NN	O	O
PU.1	NN	O	B-protein
and	NN	O	O
Ets-1	NN	O	B-protein
in	NN	O	O
nonlymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
trans-activated	NN	O	B-DNA
reporter	NN	O	I-DNA
plasmids	NN	O	I-DNA
that	NN	O	O
contained	NN	O	O
the	NN	O	O
minimal	NN	O	O
mu	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
implicate	NN	O	O
two	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
IgH	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

1	NN	O	O
,	NN	O	O
25-Dihydroxy	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
and	NN	O	O
12-O-tetradecanoyl	NN	O	O
phorbol-13-acetate	NN	O	O
synergistically	NN	O	O
induce	NN	O	O
monocytic	NN	O	B-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
:	NN	O	O
FOS	NN	O	B-protein
and	NN	O	O
RB	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

1	NN	O	O
,	NN	O	O
25-dihydroxy	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
and	NN	O	O
12-O-tetradecanoyl	NN	O	O
phorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
interact	NN	O	O
synergistically	NN	O	O
to	NN	O	O
induce	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
histiocytic	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
TPA	NN	O	O
causes	NN	O	O
an	NN	O	O
otherwise	NN	O	O
ineffective	NN	O	O
dose	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxy	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
to	NN	O	O
induce	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
simultaneous	NN	O	O
(	NN	O	O
vs.	NN	O	O
sequential	NN	O	O
)	NN	O	O
presence	NN	O	O
of	NN	O	O
both	NN	O	O
agents	NN	O	O
.	NN	O	O

The	NN	O	O
kinetics	NN	O	O
of	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
G1	NN	O	O
specific	NN	O	O
cellular	NN	O	O
response	NN	O	O
to	NN	O	O
initiate	NN	O	O
the	NN	O	O
metabolic	NN	O	O
cascade	NN	O	O
culminating	NN	O	O
in	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
occurs	NN	O	O
with	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
an	NN	O	O
accompanying	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
RB	NN	O	B-protein
protein	NN	O	I-protein
expression	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
a	NN	O	O
possible	NN	O	O
need	NN	O	O
for	NN	O	O
up-regulated	NN	O	O
RB	NN	O	B-protein
expression	NN	O	O
to	NN	O	O
maintain	NN	O	O
a	NN	O	O
given	NN	O	O
differentiated	NN	O	O
phenotype	NN	O	O
and	NN	O	O
suppress	NN	O	O
transcriptional	NN	O	B-protein
activators	NN	O	I-protein
that	NN	O	O
might	NN	O	O
typically	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
conditional	NN	O	O
GATA-2/estrogen	NN	O	B-protein
receptor	NN	O	I-protein
chimera	NN	O	I-protein
arrests	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
in	NN	O	O
a	NN	O	O
hormone-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

The	NN	O	O
GATA	NN	O	B-protein
factors	NN	O	I-protein
are	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
eukaryotes	NN	O	O
that	NN	O	O
share	NN	O	O
extensive	NN	O	O
homology	NN	O	O
in	NN	O	O
their	NN	O	O
DNA-binding	NN	O	B-protein
domains	NN	O	I-protein
.	NN	O	O

One	NN	O	O
enigmatic	NN	O	O
aspect	NN	O	O
of	NN	O	O
GATA	NN	O	B-protein
factor	NN	O	I-protein
expression	NN	O	O
is	NN	O	O
that	NN	O	O
several	NN	O	O
GATA	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
which	NN	O	O
ostensibly	NN	O	O
share	NN	O	O
the	NN	O	O
same	NN	O	O
DNA-binding	NN	O	B-protein
site	NN	O	I-protein
specificity	NN	O	O
,	NN	O	O
are	NN	O	O
coexpressed	NN	O	O
in	NN	O	O
erythroid	NN	O	O
cells	NN	O	O
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
individual	NN	O	O
GATA	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
erythropoiesis	NN	O	O
,	NN	O	O
conditional	NN	O	O
alleles	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
GATA-2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
GATA-3	NN	O	B-protein
were	NN	O	O
prepared	NN	O	O
by	NN	O	O
fusing	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
factors	NN	O	O
to	NN	O	O
the	NN	O	O
hormone-binding	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
estrogen	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
.	NN	O	O

These	NN	O	O
GATA/ER	NN	O	B-protein
chimeric	NN	O	I-protein
factors	NN	O	I-protein
were	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
hormone-inducible	NN	O	B-protein
trans-activating	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
.	NN	O	O

When	NN	O	O
stably	NN	O	O
introduced	NN	O	O
into	NN	O	O
primary	NN	O	B-cell_type
erythroblasts	NN	O	I-cell_type
or	NN	O	O
conditionally	NN	O	B-cell_line
transformed	NN	O	I-cell_line
erythroid	NN	O	I-cell_line
progenitors	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
exogenous	NN	O	O
GATA-2/ER	NN	O	B-protein
promoted	NN	O	O
proliferation	NN	O	O
and	NN	O	O
inhibited	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
in	NN	O	O
an	NN	O	O
estrogen-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

These	NN	O	O
phenotypic	NN	O	O
effects	NN	O	O
are	NN	O	O
specifically	NN	O	O
attributable	NN	O	O
to	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
ectopically	NN	O	O
expressed	NN	O	O
GATA-2/ER	NN	O	B-protein
because	NN	O	O
erythroblasts	NN	O	O
expressing	NN	O	O
exogenous	NN	O	O
GATA-2	NN	O	B-protein
are	NN	O	O
constitutively	NN	O	O
arrested	NN	O	O
in	NN	O	O
differentiation	NN	O	O
and	NN	O	O
because	NN	O	O
erythroid	NN	O	B-cell_line
progenitors	NN	O	I-cell_line
expressing	NN	O	O
either	NN	O	O
Gal/ER	NN	O	B-protein
or	NN	O	O
GATA-3/ER	NN	O	B-protein
do	NN	O	O
not	NN	O	O
display	NN	O	O
a	NN	O	O
hormone-responsive	NN	O	O
block	NN	O	O
in	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
GATA-2	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
appears	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulating	NN	O	O
the	NN	O	O
self-renewal	NN	O	O
capacity	NN	O	O
of	NN	O	O
early	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
progenitor	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Mice	NN	O	O
deficient	NN	O	O
for	NN	O	O
the	NN	O	O
55	NN	O	B-protein
kd	NN	O	I-protein
tumor	NN	O	I-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
are	NN	O	O
resistant	NN	O	O
to	NN	O	O
endotoxic	NN	O	O
shock	NN	O	O
,	NN	O	O
yet	NN	O	O
succumb	NN	O	O
to	NN	O	O
L.	NN	O	O
monocytogenes	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
multiple	NN	O	O
biological	NN	O	O
activities	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
)	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
two	NN	O	O
distinct	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
of	NN	O	O
55	NN	O	B-protein
kd	NN	O	I-protein
(	NN	O	O
TNFRp55	NN	O	B-protein
)	NN	O	O
and	NN	O	O
75	NN	O	B-protein
kd	NN	O	I-protein
(	NN	O	O
TNFRp75	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Using	NN	O	O
gene	NN	O	O
targeting	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
a	NN	O	O
TNFRp55	NN	O	B-protein
-deficient	NN	O	O
mouse	NN	O	O
strain	NN	O	O
.	NN	O	O

Cells	NN	O	O
from	NN	O	O
TNFRp55-/-mutant	NN	O	O
mice	NN	O	O
lack	NN	O	O
expression	NN	O	O
of	NN	O	O
TNFRp55	NN	O	B-protein
but	NN	O	O
display	NN	O	O
normal	NN	O	O
numbers	NN	O	O
of	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
TNFRp75	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

Thymocyte	NN	O	B-cell_type
development	NN	O	O
and	NN	O	O
lymphocyte	NN	O	B-cell_type
populations	NN	O	I-cell_type
are	NN	O	O
unaltered	NN	O	O
,	NN	O	O
and	NN	O	O
clonal	NN	O	O
deletion	NN	O	O
of	NN	O	O
potentially	NN	O	O
self-reactive	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
not	NN	O	O
impaired	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
TNF	NN	O	B-protein
signaling	NN	O	O
is	NN	O	O
largely	NN	O	O
abolished	NN	O	O
,	NN	O	O
as	NN	O	O
judged	NN	O	O
by	NN	O	O
the	NN	O	O
failure	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
TNFRp55	NN	O	B-protein
-deficient	NN	O	O
mice	NN	O	O
.	NN	O	O

The	NN	O	O
loss	NN	O	O
of	NN	O	O
TNFRp55	NN	O	B-protein
function	NN	O	O
renders	NN	O	O
mice	NN	O	O
resistant	NN	O	O
to	NN	O	O
lethal	NN	O	O
dosages	NN	O	O
of	NN	O	O
either	NN	O	O
lipopolysaccharides	NN	O	O
or	NN	O	O
S.	NN	O	B-protein
aureus	NN	O	I-protein
enterotoxin	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
TNFRp55-deficient	NN	O	O
mice	NN	O	O
are	NN	O	O
severely	NN	O	O
impaired	NN	O	O
to	NN	O	O
clear	NN	O	O
L.	NN	O	O
monocytogenes	NN	O	O
and	NN	O	O
readily	NN	O	O
succumb	NN	O	O
to	NN	O	O
infection	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
55	NN	O	B-protein
kd	NN	O	I-protein
TNFR	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
decisive	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
host	NN	O	O
's	NN	O	O
defense	NN	O	O
against	NN	O	O
microorganisms	NN	O	O
and	NN	O	O
their	NN	O	O
pathogenic	NN	O	O
factors	NN	O	O
.	NN	O	O

-DOCSTART-	O

Costimulation	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Enhanced	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
correlates	NN	O	O
with	NN	O	O
increased	NN	O	O
c-fos	NN	O	B-protein
synthesis	NN	O	O
and	NN	O	O
increased	NN	O	O
Fos	NN	O	B-protein
content	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
EC	NN	O	B-cell_type
)	NN	O	O
act	NN	O	O
as	NN	O	O
APC	NN	O	O
for	NN	O	O
resting	NN	O	B-cell_type
PBL	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
have	NN	O	O
important	NN	O	O
roles	NN	O	O
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
allograft	NN	O	O
rejection	NN	O	O
and	NN	O	O
delayed	NN	O	O
hypersensitivity	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
reported	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
umbilical	NN	O	I-cell_type
vein	NN	O	I-cell_type
EC	NN	O	I-cell_type
provide	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
to	NN	O	O
PHA-stimulated	NN	O	B-cell_type
PBL	NN	O	I-cell_type
via	NN	O	O
CD2	NN	O	B-protein
:	NN	O	I-protein
lymphocyte	NN	O	I-protein
function-associated	NN	O	I-protein
Ag-3	NN	O	I-protein
and	NN	O	O
an	NN	O	O
unidentified	NN	O	O
ligand	NN	O	O
pair	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
three-	NN	O	O
to	NN	O	O
eight-fold	NN	O	O
enhancement	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

The	NN	O	O
physiologic	NN	O	O
relevance	NN	O	O
of	NN	O	O
this	NN	O	O
increase	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
the	NN	O	O
proliferative	NN	O	O
advantage	NN	O	O
provided	NN	O	O
by	NN	O	O
EC	NN	O	B-cell_type
to	NN	O	O
PBL	NN	O	B-cell_type
suboptimally	NN	O	O
stimulated	NN	O	O
with	NN	O	O
mAb	NN	O	B-protein
OKT3	NN	O	I-protein
.	NN	O	O

We	NN	O	O
now	NN	O	O
report	NN	O	O
that	NN	O	O
EC	NN	O	B-cell_type
costimulation	NN	O	O
causes	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
increased	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
in	NN	O	O
PBL	NN	O	B-cell_type
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
EC	NN	O	B-cell_type
on	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
nuclear	NN	O	I-protein
factors	NN	O	I-protein
known	NN	O	O
to	NN	O	O
regulate	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
,	NN	O	O
including	NN	O	O
c-jun	NN	O	B-protein
and	NN	O	O
c-fos-two	NN	O	B-protein
components	NN	O	I-protein
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
,	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
and	NN	O	O
others	NN	O	O
.	NN	O	O

PBL	NN	O	B-cell_type
constitutively	NN	O	O
express	NN	O	O
c-jun	NN	O	B-protein
transcripts	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
not	NN	O	O
altered	NN	O	O
by	NN	O	O
PHA	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
or	NN	O	O
presence	NN	O	O
of	NN	O	O
EC	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
absent	NN	O	O
from	NN	O	O
resting	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
induced	NN	O	O
on	NN	O	O
PHA	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

EC	NN	O	B-cell_type
alone	NN	O	O
do	NN	O	O
not	NN	O	O
induce	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
but	NN	O	O
augment	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
PHA-activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
3-	NN	O	O
to	NN	O	O
10-fold	NN	O	O
.	NN	O	O

This	NN	O	O
effect	NN	O	O
is	NN	O	O
largely	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
CD2	NN	O	O
:	NN	O	O
lymphocyte	NN	O	O
function-associated	NN	O	O
Ag-3	NN	O	O
pathway	NN	O	O
.	NN	O	O

Gel-shift	NN	O	O
analysis	NN	O	O
reveals	NN	O	O
the	NN	O	O
constitutive	NN	O	O
presence	NN	O	O
of	NN	O	O
nuclear	NN	O	O
factors	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
PBL	NN	O	I-cell_type
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
AP-1	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
contain	NN	O	O
immunoreactive	NN	O	O
c-Jun	NN	O	B-protein
but	NN	O	O
not	NN	O	O
c-Fos	NN	O	B-protein
protein	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
AP-1	NN	O	B-protein
from	NN	O	O
PHA-activated	NN	O	B-cell_type
cells	NN	O	I-cell_type
contains	NN	O	O
c-Jun	NN	O	B-protein
and	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
.	NN	O	O

Strikingly	NN	O	O
,	NN	O	O
costimulation	NN	O	O
with	NN	O	O
EC	NN	O	B-cell_type
results	NN	O	O
in	NN	O	O
a	NN	O	O
dramatic	NN	O	O
increase	NN	O	O
(	NN	O	O
up	NN	O	O
to	NN	O	O
15-fold	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
c-Fos	NN	O	B-protein
content	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Levels	NN	O	O
of	NN	O	O
other	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
regulation	NN	O	O
were	NN	O	O
not	NN	O	O
altered	NN	O	O
by	NN	O	O
EC	NN	O	B-cell_type
,	NN	O	O
although	NN	O	O
NFAT-DNA	NN	O	B-protein
complexes	NN	O	I-protein
migrated	NN	O	O
at	NN	O	O
a	NN	O	O
slightly	NN	O	O
different	NN	O	O
mobility	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
composition	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
key	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
for	NN	O	O
increasing	NN	O	O
IL-2	NN	O	B-protein
transcription	NN	O	O
and	NN	O	O
may	NN	O	O
underlie	NN	O	O
the	NN	O	O
phenomenon	NN	O	O
of	NN	O	O
costimulation	NN	O	O
by	NN	O	O
EC	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
zeta	NN	O	I-DNA
2	NN	O	I-DNA
globin	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
alpha	NN	O	B-DNA
globin	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
HS-40	NN	O	B-DNA
)	NN	O	O
:	NN	O	O
functional	NN	O	O
role	NN	O	O
of	NN	O	O
specific	NN	O	O
nuclear	NN	O	B-protein
factor-DNA	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

We	NN	O	O
studied	NN	O	O
the	NN	O	O
functional	NN	O	O
interaction	NN	O	O
between	NN	O	O
human	NN	O	B-DNA
embryonic	NN	O	I-DNA
zeta	NN	O	I-DNA
2	NN	O	I-DNA
globin	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
alpha	NN	O	B-DNA
globin	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
HS-40	NN	O	B-DNA
)	NN	O	O
located	NN	O	O
40	NN	O	B-DNA
kb	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
zeta	NN	O	B-DNA
2	NN	O	I-DNA
globin	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
was	NN	O	O
shown	NN	O	O
by	NN	O	O
transient	NN	O	O
expression	NN	O	O
assay	NN	O	O
that	NN	O	O
HS-40	NN	O	B-DNA
behaved	NN	O	O
as	NN	O	O
an	NN	O	O
authentic	NN	O	O
enhancer	NN	O	B-DNA
for	NN	O	O
high-level	NN	O	O
zeta	NN	O	B-DNA
2	NN	O	I-DNA
globin	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
an	NN	O	O
erythroid	NN	O	O
cell	NN	O	O
line	NN	O	O
of	NN	O	O
embryonic	NN	O	O
and/or	NN	O	O
fetal	NN	O	O
origin	NN	O	O
.	NN	O	O

Although	NN	O	O
sequences	NN	O	O
located	NN	O	O
between	NN	O	O
-559	NN	O	O
and	NN	O	O
-88	NN	O	O
of	NN	O	O
the	NN	O	O
zeta	NN	O	B-DNA
2	NN	O	I-DNA
globin	NN	O	I-DNA
gene	NN	O	I-DNA
were	NN	O	O
dispensable	NN	O	O
for	NN	O	O
its	NN	O	O
expression	NN	O	O
on	NN	O	O
enhancerless	NN	O	B-DNA
plasmids	NN	O	I-DNA
,	NN	O	O
they	NN	O	O
were	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
HS-40	NN	O	B-DNA
enhancer	NN	O	I-DNA
-mediated	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
zeta	NN	O	B-DNA
2	NN	O	I-DNA
globin	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
this	NN	O	O
HS-40	NN	O	B-DNA
enhancer-	NN	O	O
zeta	NN	O	B-DNA
2	NN	O	I-DNA
globin	NN	O	I-DNA
promoter	NN	O	I-DNA
interaction	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
two	NN	O	O
GATA-1	NN	O	O
factor	NN	O	O
binding	NN	O	O
motifs	NN	O	O
located	NN	O	O
at	NN	O	O
-230	NN	O	O
and	NN	O	O
-104	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
functional	NN	O	O
domains	NN	O	O
of	NN	O	O
HS-40	NN	O	B-DNA
were	NN	O	O
also	NN	O	O
mapped	NN	O	O
.	NN	O	O

Bal	NN	O	O
31	NN	O	O
deletion	NN	O	O
mapping	NN	O	O
data	NN	O	O
suggested	NN	O	O
that	NN	O	O
one	NN	O	O
GATA-1	NN	O	O
motif	NN	O	O
,	NN	O	O
one	NN	O	O
GT	NN	O	O
motif	NN	O	O
,	NN	O	O
and	NN	O	O
two	NN	O	O
NF-E2/AP1	NN	O	O
motifs	NN	O	O
together	NN	O	O
formed	NN	O	O
the	NN	O	O
functional	NN	O	O
core	NN	O	O
of	NN	O	O
HS-40	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
erythroid-specific	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
zeta	NN	O	B-DNA
2	NN	O	I-DNA
globin	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
further	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
function	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
NF-E2/AP1	NN	O	O
motifs	NN	O	O
of	NN	O	O
HS-40	NN	O	B-DNA
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
its	NN	O	O
binding	NN	O	O
to	NN	O	O
NF-E2	NN	O	B-protein
but	NN	O	O
not	NN	O	O
AP1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
did	NN	O	O
genomic	NN	O	O
footprinting	NN	O	O
of	NN	O	O
the	NN	O	O
HS-40	NN	O	B-DNA
enhancer	NN	O	I-DNA
region	NN	O	I-DNA
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
adult	NN	O	B-cell_line
nucleated	NN	O	I-cell_line
erythroblasts	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
different	NN	O	O
nonerythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

All	NN	O	O
sequence	NN	O	O
motifs	NN	O	O
within	NN	O	O
the	NN	O	O
functional	NN	O	O
core	NN	O	O
of	NN	O	O
HS-40	NN	O	B-DNA
,	NN	O	O
as	NN	O	O
mapped	NN	O	O
by	NN	O	O
transient	NN	O	O
expression	NN	O	O
analysis	NN	O	O
,	NN	O	O
appeared	NN	O	O
to	NN	O	O
bind	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
in	NN	O	O
living	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
nonerythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
only	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
apparently	NN	O	O
nonfunctional	NN	O	B-DNA
sequence	NN	O	I-DNA
motifs	NN	O	I-DNA
was	NN	O	O
bound	NN	O	O
with	NN	O	O
factors	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

In	NN	O	O
comparison	NN	O	O
to	NN	O	O
K562	NN	O	B-cell_line
,	NN	O	O
nucleated	NN	O	B-cell_line
erythroblasts	NN	O	I-cell_line
from	NN	O	O
adult	NN	O	O
human	NN	O	O
bone	NN	O	O
marrow	NN	O	O
exhibited	NN	O	O
a	NN	O	O
similar	NN	O	O
but	NN	O	O
nonidentical	NN	O	O
pattern	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
in	NN	O	O
vivo	NN	O	O
at	NN	O	O
the	NN	O	O
HS-40	NN	O	B-DNA
region	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
embryonic	NN	O	I-DNA
zeta	NN	O	I-DNA
2	NN	O	I-DNA
globin	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
fetal/adult	NN	O	B-DNA
alpha	NN	O	I-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
erythroid	NN	O	O
cell-specific	NN	O	O
and	NN	O	O
developmental	NN	O	B-protein
stage-specific	NN	O	I-protein
nuclear	NN	O	I-protein
factor-DNA	NN	O	I-protein
complexes	NN	O	I-protein
which	NN	O	O
form	NN	O	O
at	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
(	NN	O	O
HS-40	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
the	NN	O	O
globin	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
pyruvate	NN	O	B-DNA
kinase	NN	O	I-DNA
erythroid-specific	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Mammal	NN	O	B-protein
pyruvate	NN	O	I-protein
kinases	NN	O	I-protein
are	NN	O	O
encoded	NN	O	O
by	NN	O	O
two	NN	O	O
genes	NN	O	O
.	NN	O	O

The	NN	O	O
L	NN	O	B-DNA
gene	NN	O	I-DNA
produces	NN	O	O
the	NN	O	O
erythroid	NN	O	B-protein
(	NN	O	O
R-PK	NN	O	B-protein
)	NN	O	O
or	NN	O	O
the	NN	O	O
hepatic	NN	O	B-protein
(	NN	O	I-protein
L-PK	NN	O	I-protein
)	NN	O	I-protein
isozymes	NN	O	I-protein
by	NN	O	O
the	NN	O	O
alternative	NN	O	O
use	NN	O	O
of	NN	O	O
two	NN	O	O
promoters	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
cis-	NN	O	B-DNA
and	NN	O	I-DNA
trans-acting	NN	O	I-DNA
elements	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
L	NN	O	B-DNA
gene	NN	O	I-DNA
erythroid	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
R-PK	NN	O	B-DNA
DNA	NN	O	I-DNA
fragment	NN	O	I-DNA
extending	NN	O	O
from	NN	O	O
-870	NN	O	O
to	NN	O	O
+54	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
cap	NN	O	B-DNA
site	NN	O	I-DNA
confers	NN	O	O
erythroid	NN	O	O
specificity	NN	O	O
to	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Within	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
we	NN	O	O
define	NN	O	O
a	NN	O	O
minimal	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
-62	NN	O	O
to	NN	O	O
+54	NN	O	O
)	NN	O	O
that	NN	O	O
displays	NN	O	O
erythroid-specific	NN	O	O
activity	NN	O	O
and	NN	O	O
contains	NN	O	O
two	NN	O	O
DNA	NN	O	O
binding	NN	O	O
sites	NN	O	O
.	NN	O	O

One	NN	O	O
,	NN	O	O
located	NN	O	O
at	NN	O	O
-50	NN	O	O
,	NN	O	O
binds	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
CCACC/Sp1	NN	O	B-protein
family	NN	O	I-protein
and	NN	O	O
the	NN	O	O
other	NN	O	O
,	NN	O	O
located	NN	O	O
at	NN	O	O
-20	NN	O	O
,	NN	O	O
binds	NN	O	O
the	NN	O	O
erythroid	NN	O	B-protein
factor	NN	O	I-protein
GATA-1	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
the	NN	O	O
-20	NN	O	O
GATA	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
AGATAA	NN	O	O
)	NN	O	O
is	NN	O	O
also	NN	O	O
a	NN	O	O
potential	NN	O	O
TFIID	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
it	NN	O	O
does	NN	O	O
not	NN	O	O
bind	NN	O	O
TFIID	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
this	NN	O	O
GATA	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
by	NN	O	O
a	NN	O	O
canonical	NN	O	O
TFIID	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
suppresses	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Mutations	NN	O	O
and	NN	O	O
deletions	NN	O	O
of	NN	O	O
both	NN	O	O
sites	NN	O	O
indicate	NN	O	O
that	NN	O	O
only	NN	O	O
the	NN	O	O
association	NN	O	O
of	NN	O	O
CCACC/Sp1	NN	O	B-protein
and	NN	O	O
GATA	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
can	NN	O	O
drive	NN	O	O
efficient	NN	O	O
and	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
R-PK	NN	O	B-protein
minimal	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
by	NN	O	O
co-transfection	NN	O	O
experiments	NN	O	O
,	NN	O	O
we	NN	O	O
study	NN	O	O
the	NN	O	O
elements	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
hGATA-1	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
R-PK	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Protease	NN	O	O
treatment	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
distinguishes	NN	O	O
between	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
MHC	NN	O	I-protein
X1	NN	O	I-protein
box	NN	O	I-protein
DNA-binding	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
wild-type	NN	O	B-cell_line
and	NN	O	I-cell_line
class	NN	O	I-cell_line
II-deficient	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
X	NN	O	B-DNA
box	NN	O	I-DNA
region	NN	O	I-DNA
is	NN	O	O
critical	NN	O	O
for	NN	O	O
directing	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
major	NN	O	I-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
several	NN	O	O
class	NN	O	B-protein
II	NN	O	I-protein
promoter-specific	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
factors	NN	O	I-protein
have	NN	O	O
been	NN	O	O
described	NN	O	O
,	NN	O	O
only	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
box	NN	O	I-DNA
region	NN	O	I-DNA
factor	NN	O	O
,	NN	O	O
RFX	NN	O	B-protein
,	NN	O	O
shows	NN	O	O
a	NN	O	O
genetic	NN	O	O
correlation	NN	O	O
with	NN	O	O
class	NN	O	O
II	NN	O	O
expression	NN	O	O
,	NN	O	O
being	NN	O	O
deficient	NN	O	O
in	NN	O	O
some	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
class	NN	O	O
II-deficient	NN	O	O
congenital	NN	O	O
immunodeficiency	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
evaluate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
X	NN	O	B-protein
box	NN	O	I-protein
DNA-binding	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
box	NN	O	I-DNA
region	NN	O	I-DNA
was	NN	O	O
examined	NN	O	O
in	NN	O	O
both	NN	O	O
class	NN	O	B-cell_line
II-positive	NN	O	I-cell_line
and	NN	O	O
-negative	NN	O	B-cell_line
lymphoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
wild-type	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Raji	NN	O	I-cell_line
,	NN	O	O
two	NN	O	B-cell_line
class	NN	O	I-cell_line
II	NN	O	I-cell_line
transcriptional	NN	O	I-cell_line
mutant	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
SJO	NN	O	B-cell_line
and	NN	O	O
RJ2.2.5	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
Jurkat	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
class	NN	O	B-cell_line
II	NN	O	I-cell_line
negative	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
were	NN	O	O
examined	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
wild-type	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
neither	NN	O	O
of	NN	O	O
the	NN	O	O
class	NN	O	B-cell_line
II	NN	O	I-cell_line
mutant	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
could	NN	O	O
use	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
box	NN	O	I-DNA
region	NN	O	I-DNA
to	NN	O	O
direct	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
X	NN	O	O
box-dependent	NN	O	O
transcriptional	NN	O	O
pathway	NN	O	O
is	NN	O	O
defective	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
X1	NN	O	B-protein
box	NN	O	I-protein
DNA-binding	NN	O	I-protein
protein	NN	O	I-protein
RFX	NN	O	B-protein
was	NN	O	O
examined	NN	O	O
and	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
present	NN	O	O
in	NN	O	O
wild-type	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
mutant	NN	O	O
RJ2.2.5	NN	O	B-cell_line
but	NN	O	O
was	NN	O	O
absent	NN	O	O
in	NN	O	O
SJO	NN	O	B-cell_line
and	NN	O	O
Jurkat	NN	O	B-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
other	NN	O	O
X1	NN	O	O
box-specific	NN	O	O
activities	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
all	NN	O	O
these	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
these	NN	O	O
different	NN	O	O
X1	NN	O	O
box	NN	O	O
activities	NN	O	O
represented	NN	O	O
distinct	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
or	NN	O	O
multimeric	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
,	NN	O	O
protease	NN	O	B-protein
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
crude	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
followed	NN	O	O
by	NN	O	O
DNA-binding	NN	O	O
assays	NN	O	O
were	NN	O	O
carried	NN	O	O
out	NN	O	O
and	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
B	NN	O	O
cell	NN	O	O
extracts	NN	O	O
contain	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
X1-specific	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
cleaved	NN	O	O
products	NN	O	O
(	NN	O	O
band	NN	O	O
1	NN	O	O
pk	NN	O	O
)	NN	O	O
correlates	NN	O	O
with	NN	O	O
RFX	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
comparison	NN	O	O
with	NN	O	O
protease-treated	NN	O	O
extracts	NN	O	O
prepared	NN	O	O
from	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
demonstrated	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
band	NN	O	O
1pk	NN	O	O
activity	NN	O	O
despite	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
native	NN	O	O
RFX	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
protease	NN	O	O
treatment	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
SJO	NN	O	B-cell_line
extracts	NN	O	O
showed	NN	O	O
no	NN	O	O
detectable	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
band	NN	O	O
1pk	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
multiple	NN	O	O
X1	NN	O	O
box-specific	NN	O	O
DNA-binding	NN	O	O
activities	NN	O	O
exist	NN	O	O
in	NN	O	O
all	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
actively	NN	O	O
binding	NN	O	O
RFX	NN	O	B-protein
species	NN	O	O
correlates	NN	O	O
with	NN	O	O
class	NN	O	O
II	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
immunodeficiency	NN	O	O
viruses	NN	O	O
containing	NN	O	O
heterologous	NN	O	B-DNA
enhancer/promoters	NN	O	I-DNA
are	NN	O	O
replication	NN	O	O
competent	NN	O	O
and	NN	O	O
exhibit	NN	O	O
different	NN	O	O
lymphocyte	NN	O	O
tropisms	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
HIV	NN	O	I-DNA
)	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
contains	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Sp1	NN	O	I-protein
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
highly	NN	O	O
conserved	NN	O	O
in	NN	O	O
HIV	NN	O	O
and	NN	O	O
simian	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
isolates	NN	O	O
.	NN	O	O

To	NN	O	O
delineate	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
these	NN	O	O
motifs	NN	O	O
on	NN	O	O
the	NN	O	O
replicative	NN	O	O
capacity	NN	O	O
of	NN	O	O
HIV	NN	O	O
and	NN	O	O
to	NN	O	O
explore	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
extending	NN	O	O
the	NN	O	O
virus	NN	O	O
host	NN	O	O
range	NN	O	O
,	NN	O	O
known	NN	O	O
heterologous	NN	O	B-DNA
enhancer/promoters	NN	O	I-DNA
were	NN	O	O
inserted	NN	O	O
into	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
in	NN	O	O
place	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
Sp1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
these	NN	O	O
substitutions	NN	O	O
on	NN	O	O
viral	NN	O	O
replication	NN	O	O
in	NN	O	O
transfected	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
on	NN	O	O
HIV	NN	O	O
infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
or	NN	O	O
continuous	NN	O	B-cell_line
T-leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
evaluated	NN	O	O
.	NN	O	O

HIVs	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B/Sp1	NN	O	I-DNA
enhancer	NN	O	I-DNA
plus	NN	O	O
the	NN	O	O
downstream	NN	O	B-DNA
TATA	NN	O	I-DNA
element	NN	O	I-DNA
were	NN	O	O
replaced	NN	O	O
with	NN	O	O
heterologous	NN	O	B-DNA
enhancer/promoters	NN	O	I-DNA
were	NN	O	O
also	NN	O	O
constructed	NN	O	O
.	NN	O	O

Viruses	NN	O	O
containing	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
cytomegalovirus	NN	O	I-DNA
immediate-early	NN	O	I-DNA
enhancer	NN	O	I-DNA
exhibited	NN	O	O
infectious	NN	O	O
kinetics	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
wild-type	NN	O	O
HIV	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
AA2	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
replicated	NN	O	O
less	NN	O	O
efficiently	NN	O	O
in	NN	O	O
H9	NN	O	B-cell_line
and	NN	O	I-cell_line
CEM	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
heterologous	NN	O	B-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
are	NN	O	O
capable	NN	O	O
of	NN	O	O
restoring	NN	O	O
Tat	NN	O	B-protein
responsiveness	NN	O	O
to	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
directing	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
infectious	NN	O	O
progeny	NN	O	O
virions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tyrosine	NN	O	O
phosphorylation	NN	O	O
is	NN	O	O
a	NN	O	O
mandatory	NN	O	O
proximal	NN	O	O
step	NN	O	O
in	NN	O	O
radiation-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
signaling	NN	O	O
pathway	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B-lymphocyte	NN	O	I-cell_type
precursors	NN	O	I-cell_type
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
Proc	NN	O	O
Natl	NN	O	O
Acad	NN	O	O
Sci	NN	O	O
U	NN	O	O
S	NN	O	O
A	NN	O	O
1993	NN	O	O
Apr	NN	O	O
15	NN	O	O
;	NN	O	O
90	NN	O	O
(	NN	O	O
8	NN	O	O
)	NN	O	O
:	NN	O	O
3775	NN	O	O
]	NN	O	O

Ionizing	NN	O	O
radiation	NN	O	O
triggers	NN	O	O
a	NN	O	O
signal	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B-lymphocyte	NN	O	I-cell_type
precursors	NN	O	I-cell_type
that	NN	O	O
is	NN	O	O
intimately	NN	O	O
linked	NN	O	O
to	NN	O	O
an	NN	O	O
active	NN	O	O
protein-tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
regulatory	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
B-lymphocyte	NN	O	B-cell_type
precursors	NN	O	I-cell_type
,	NN	O	O
irradiation	NN	O	O
with	NN	O	O
gamma-rays	NN	O	O
leads	NN	O	O
to	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
stimulation	NN	O	O
of	NN	O	O
phosphatidylinositol	NN	O	O
turnover	NN	O	O
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
downstream	NN	O	O
activation	NN	O	O
by	NN	O	O
covalent	NN	O	O
modification	NN	O	O
of	NN	O	O
multiple	NN	O	O
serine-specific	NN	O	B-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
,	NN	O	O
including	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
;	NN	O	O
and	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
activation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

All	NN	O	O
of	NN	O	O
the	NN	O	O
radiation-induced	NN	O	O
signals	NN	O	O
were	NN	O	O
effectively	NN	O	O
prevented	NN	O	O
by	NN	O	O
the	NN	O	O
protein-tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
inhibitors	NN	O	I-protein
genistein	NN	O	I-protein
and	NN	O	I-protein
herbimycin	NN	O	I-protein
A	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
and	NN	O	O
perhaps	NN	O	O
mandatory	NN	O	O
proximal	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
signaling	NN	O	O
cascade	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B-lymphocyte	NN	O	I-cell_type
precursors	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
report	NN	O	O
expands	NN	O	O
current	NN	O	O
knowledge	NN	O	O
of	NN	O	O
the	NN	O	O
radiation-induced	NN	O	O
signaling	NN	O	O
cascade	NN	O	O
by	NN	O	O
clarifying	NN	O	O
the	NN	O	O
chronological	NN	O	O
sequence	NN	O	O
of	NN	O	O
biochemical	NN	O	O
events	NN	O	O
that	NN	O	O
follow	NN	O	O
irradiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Detection	NN	O	O
of	NN	O	O
adenovirus	NN	O	B-DNA
DNA	NN	O	I-DNA
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
assay	NN	O	O
.	NN	O	O

Adenovirus	NN	O	O
can	NN	O	O
establish	NN	O	O
persistent	NN	O	O
infections	NN	O	O
which	NN	O	O
may	NN	O	O
reactivate	NN	O	O
and	NN	O	O
cause	NN	O	O
disease	NN	O	O
in	NN	O	O
immunocompromised	NN	O	O
hosts	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
have	NN	O	O
been	NN	O	O
postulated	NN	O	O
to	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
site	NN	O	O
of	NN	O	O
adenoviral	NN	O	O
persistence	NN	O	O
based	NN	O	O
upon	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
isolate	NN	O	O
adenovirus	NN	O	O
from	NN	O	O
tonsils	NN	O	O
and	NN	O	O
to	NN	O	O
detect	NN	O	O
adenovirus	NN	O	B-DNA
DNA	NN	O	I-DNA
by	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
PBMC	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

To	NN	O	O
test	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
a	NN	O	O
more	NN	O	O
sensitive	NN	O	O
and	NN	O	O
specific	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
assay	NN	O	O
was	NN	O	O
developed	NN	O	O
to	NN	O	O
detect	NN	O	O
adenovirus	NN	O	B-DNA
DNA	NN	O	I-DNA
.	NN	O	O

Two	NN	O	O
sets	NN	O	O
of	NN	O	O
nested	NN	O	B-DNA
primers	NN	O	I-DNA
were	NN	O	O
designed	NN	O	O
to	NN	O	O
conserved	NN	O	O
sequences	NN	O	O
in	NN	O	O
the	NN	O	O
adenovirus	NN	O	B-DNA
E1A	NN	O	I-DNA
and	NN	O	I-DNA
hexon	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
E1A	NN	O	B-DNA
and	NN	O	I-DNA
hexon	NN	O	I-DNA
primers	NN	O	I-DNA
amplified	NN	O	O
DNA	NN	O	O
from	NN	O	O
representative	NN	O	O
adenoviral	NN	O	O
serotypes	NN	O	O
in	NN	O	O
all	NN	O	O
six	NN	O	O
adenoviral	NN	O	O
groups	NN	O	O
(	NN	O	O
A-F	NN	O	O
)	NN	O	O
.	NN	O	O

Both	NN	O	O
primers	NN	O	O
detected	NN	O	O
a	NN	O	O
single	NN	O	O
copy	NN	O	O
of	NN	O	O
the	NN	O	O
adenovirus	NN	O	B-DNA
type	NN	O	I-DNA
2	NN	O	I-DNA
genome	NN	O	I-DNA
but	NN	O	O
were	NN	O	O
less	NN	O	O
sensitive	NN	O	O
for	NN	O	O
the	NN	O	O
group	NN	O	O
B	NN	O	O
type	NN	O	O
35	NN	O	O
.	NN	O	O

None	NN	O	O
of	NN	O	O
33	NN	O	O
PBMC	NN	O	B-cell_type
specimens	NN	O	O
from	NN	O	O
healthy	NN	O	O
adults	NN	O	O
and	NN	O	O
only	NN	O	O
one	NN	O	O
of	NN	O	O
40	NN	O	O
pediatric	NN	O	O
samples	NN	O	O
was	NN	O	O
positive	NN	O	O
(	NN	O	O
at	NN	O	O
a	NN	O	O
low	NN	O	O
level	NN	O	O
)	NN	O	O
for	NN	O	O
adenovirus	NN	O	B-DNA
DNA	NN	O	I-DNA
by	NN	O	O
nested	NN	O	O
PCR	NN	O	O
assay	NN	O	O
.	NN	O	O

In	NN	O	O
comparison	NN	O	O
,	NN	O	O
PBMC	NN	O	B-cell_type
from	NN	O	O
two	NN	O	O
children	NN	O	O
with	NN	O	O
fatal	NN	O	O
adenoviral	NN	O	O
infection	NN	O	O
were	NN	O	O
both	NN	O	O
strongly	NN	O	O
positive	NN	O	O
for	NN	O	O
adenovirus	NN	O	B-DNA
DNA	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
a	NN	O	O
previous	NN	O	O
study	NN	O	O
,	NN	O	O
PBMC	NN	O	B-cell_type
are	NN	O	O
not	NN	O	O
a	NN	O	O
common	NN	O	O
site	NN	O	O
of	NN	O	O
persistent	NN	O	O
group	NN	O	O
C	NN	O	O
adenoviral	NN	O	O
infection	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
assay	NN	O	O
of	NN	O	O
PBMC	NN	O	B-cell_type
by	NN	O	O
the	NN	O	O
adenovirus-specific	NN	O	O
PCR	NN	O	O
may	NN	O	O
help	NN	O	O
detect	NN	O	O
early	NN	O	O
invasive	NN	O	O
disease	NN	O	O
and	NN	O	O
warrants	NN	O	O
further	NN	O	O
evaluation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
signal	NN	O	O
transduction	NN	O	O
in	NN	O	O
HIV-1-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
To	NN	O	O
examine	NN	O	O
the	NN	O	O
inter-relationship	NN	O	O
between	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
and	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
receptors	NN	O	O
for	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TNF	NN	O	I-protein
)	NN	O	I-protein
-alpha	NN	O	I-protein
,	NN	O	O
an	NN	O	O
immunoregulatory	NN	O	O
cytokine	NN	O	O
that	NN	O	O
can	NN	O	O
enhance	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

DESIGN	NN	O	O
:	NN	O	O
Infected	NN	O	O
promyelocytic	NN	O	B-cell_type
and	NN	O	O
promonocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
examined	NN	O	O
because	NN	O	O
they	NN	O	O
normally	NN	O	O
express	NN	O	O
both	NN	O	O
types	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
TNF	NN	O	B-protein
receptor	NN	O	O
surface	NN	O	O
expression	NN	O	O
was	NN	O	O
determined	NN	O	O
by	NN	O	O
specific	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
recognition	NN	O	O
and	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
,	NN	O	O
and	NN	O	O
signal	NN	O	O
transduction	NN	O	O
was	NN	O	O
detected	NN	O	O
by	NN	O	O
gel	NN	O	O
shift	NN	O	O
analysis	NN	O	O
.	NN	O	O

HIV-1	NN	O	O
activation	NN	O	O
and	NN	O	O
expression	NN	O	O
was	NN	O	O
quantitated	NN	O	O
by	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
assay	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
In	NN	O	O
the	NN	O	O
OM-10.1	NN	O	O
promyelocytic	NN	O	O
model	NN	O	O
of	NN	O	O
chronic	NN	O	O
infection	NN	O	O
,	NN	O	O
TNF-alpha-induced	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
also	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
substantial	NN	O	O
increase	NN	O	O
in	NN	O	O
75	NN	O	B-protein
kd	NN	O	I-protein
TNF	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TR75	NN	O	B-protein
)	NN	O	O
expression	NN	O	O
although	NN	O	O
55	NN	O	B-protein
kD	NN	O	I-protein
TNF	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TR55	NN	O	B-protein
)	NN	O	O
levels	NN	O	O
were	NN	O	O
not	NN	O	O
dramatically	NN	O	O
altered	NN	O	O
.	NN	O	O

A	NN	O	O
series	NN	O	O
of	NN	O	O
uninfected	NN	O	B-cell_line
parental	NN	O	I-cell_line
HL-60	NN	O	I-cell_line
subclones	NN	O	I-cell_line
all	NN	O	O
reduced	NN	O	O
TR75	NN	O	B-protein
surface	NN	O	O
expression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TNF-alpha	NN	O	B-protein
treatment	NN	O	O
.	NN	O	O

Enhanced	NN	O	O
TR75	NN	O	B-protein
expression	NN	O	O
on	NN	O	O
OM-10.1	NN	O	O
cells	NN	O	O
followed	NN	O	O
the	NN	O	O
same	NN	O	O
TNF-alpha	NN	O	B-protein
-dose	NN	O	O
dependency	NN	O	O
as	NN	O	O
that	NN	O	O
observed	NN	O	O
for	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
.	NN	O	O

An	NN	O	O
increase	NN	O	O
in	NN	O	O
TR75	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
also	NN	O	O
evident	NN	O	O
during	NN	O	O
the	NN	O	O
peak	NN	O	O
of	NN	O	O
an	NN	O	O
acute	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
promyelocytes	NN	O	I-cell_line
.	NN	O	O

Although	NN	O	O
TR55	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
unaltered	NN	O	O
during	NN	O	O
TNF-alpha	NN	O	B-protein
-induced	NN	O	O
HIV	NN	O	O
activation	NN	O	O
,	NN	O	O
this	NN	O	O
receptor	NN	O	O
was	NN	O	O
still	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
viral	NN	O	O
activation	NN	O	O
process	NN	O	O
.	NN	O	O

Antibody	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
TR55	NN	O	B-protein
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
TNF-alpha	NN	O	I-protein
,	NN	O	O
induced	NN	O	O
maximal	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
,	NN	O	O
an	NN	O	O
up-modulation	NN	O	O
of	NN	O	O
surface	NN	O	O
TR75	NN	O	B-protein
,	NN	O	O
and	NN	O	O
nuclear	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
OM-10.1	NN	O	O
cultures	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
this	NN	O	O
was	NN	O	O
the	NN	O	O
case	NN	O	O
even	NN	O	O
when	NN	O	O
an	NN	O	O
antagonistic	NN	O	B-protein
anti-TR55	NN	O	I-protein
antibody	NN	O	I-protein
was	NN	O	O
used	NN	O	O
.	NN	O	O

Anti-TR55	NN	O	B-protein
antibody	NN	O	I-protein
cross-linking	NN	O	O
in	NN	O	O
chronically	NN	O	B-cell_line
infected	NN	O	I-cell_line
U1	NN	O	I-cell_line
promonocytic	NN	O	I-cell_line
cultures	NN	O	I-cell_line
could	NN	O	O
only	NN	O	O
partially	NN	O	O
substitute	NN	O	O
for	NN	O	O
TNF-alpha-induced	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Our	NN	O	O
results	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
can	NN	O	O
selectively	NN	O	O
influence	NN	O	O
the	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
potentially	NN	O	O
influencing	NN	O	O
its	NN	O	O
own	NN	O	O
expression	NN	O	O
and	NN	O	O
altering	NN	O	O
normal	NN	O	O
immunoregulatory	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

-DOCSTART-	O

Comparative	NN	O	O
mapping	NN	O	O
of	NN	O	O
SRY	NN	O	B-DNA
in	NN	O	O
the	NN	O	O
great	NN	O	O
apes	NN	O	O
.	NN	O	O

Cytogenetic	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
primate	NN	O	B-DNA
Y	NN	O	I-DNA
chromosomes	NN	O	I-DNA
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
extensive	NN	O	O
rearrangements	NN	O	O
have	NN	O	O
occurred	NN	O	O
during	NN	O	O
evolution	NN	O	O
of	NN	O	O
the	NN	O	O
great	NN	O	O
apes	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
to	NN	O	O
define	NN	O	O
these	NN	O	O
rearrangements	NN	O	O
at	NN	O	O
the	NN	O	O
molecular	NN	O	O
level	NN	O	O
.	NN	O	O

pHU-14	NN	O	B-DNA
,	NN	O	O
a	NN	O	O
probe	NN	O	O
including	NN	O	O
sequences	NN	O	O
from	NN	O	O
the	NN	O	O
sex	NN	O	B-DNA
determining	NN	O	I-DNA
gene	NN	O	I-DNA
SRY	NN	O	I-DNA
,	NN	O	O
hybridizes	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
early	NN	O	O
replicating	NN	O	O
pseudoautosomal	NN	O	O
segment	NN	O	O
in	NN	O	O
a	NN	O	O
telomeric	NN	O	O
or	NN	O	O
subtelomeric	NN	O	O
position	NN	O	O
of	NN	O	O
the	NN	O	O
Y	NN	O	B-DNA
chromosomes	NN	O	I-DNA
of	NN	O	O
all	NN	O	O
great	NN	O	O
apes	NN	O	O
.	NN	O	O

The	NN	O	O
low	NN	O	B-DNA
copy	NN	O	I-DNA
repeat	NN	O	I-DNA
detected	NN	O	O
by	NN	O	O
the	NN	O	O
probe	NN	O	O
Fr35-II	NN	O	B-DNA
is	NN	O	O
obviously	NN	O	O
included	NN	O	O
in	NN	O	O
Y	NN	O	O
chromosomal	NN	O	O
rearrangements	NN	O	O
during	NN	O	O
hominid	NN	O	O
evolution	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
,	NN	O	O
combined	NN	O	O
with	NN	O	O
previous	NN	O	O
studies	NN	O	O
,	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
Y	NN	O	O
chromosome	NN	O	O
in	NN	O	O
great	NN	O	O
apes	NN	O	O
has	NN	O	O
a	NN	O	O
conserved	NN	O	O
region	NN	O	O
including	NN	O	O
the	NN	O	O
pseudoautosomal	NN	O	B-DNA
region	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
testis-determining	NN	O	O
region	NN	O	O
.	NN	O	O

The	NN	O	O
rest	NN	O	O
of	NN	O	O
the	NN	O	O
Y	NN	O	B-DNA
chromosome	NN	O	I-DNA
has	NN	O	O
undergone	NN	O	O
several	NN	O	O
rearrangements	NN	O	O
in	NN	O	O
the	NN	O	O
different	NN	O	O
great	NN	O	O
apes	NN	O	O
.	NN	O	O

-DOCSTART-	O

Calcium	NN	O	O
dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
by	NN	O	O
p59fyn	NN	O	B-protein
.	NN	O	O

A	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
a	NN	O	O
T-cell	NN	O	B-DNA
antigen	NN	O	I-DNA
receptor	NN	O	I-DNA
element	NN	O	I-DNA
was	NN	O	O
activated	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
triggering	NN	O	O
or	NN	O	O
by	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
,	NN	O	O
which	NN	O	O
activates	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
.	NN	O	O

Both	NN	O	O
these	NN	O	O
signals	NN	O	O
were	NN	O	O
necessary	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
co-transfected	NN	O	O
with	NN	O	O
a	NN	O	O
construct	NN	O	O
capable	NN	O	O
of	NN	O	O
overexpressing	NN	O	O
the	NN	O	O
tyrosine	NN	O	B-protein
kinase	NN	O	I-protein
p59fyn	NN	O	B-protein
,	NN	O	O
the	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
activated	NN	O	O
by	NN	O	O
PMA	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

Thus	NN	O	O
p59fyn	NN	O	B-protein
could	NN	O	O
replace	NN	O	O
the	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
but	NN	O	O
not	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
.	NN	O	O

The	NN	O	O
activation	NN	O	O
by	NN	O	O
p59fyn	NN	O	B-protein
plus	NN	O	O
PMA	NN	O	B-protein
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
EGTA	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
immunosuppressant	NN	O	O
drug	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

-DOCSTART-	O

Dependence	NN	O	O
for	NN	O	O
the	NN	O	O
proliferative	NN	O	O
response	NN	O	O
to	NN	O	O
erythropoietin	NN	O	B-protein
on	NN	O	O
an	NN	O	O
established	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
program	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
hematopoietic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
UT-7	NN	O	B-cell_line
.	NN	O	O

Erythroid	NN	O	O
differentiation	NN	O	O
involves	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
erythroid-specific	NN	O	O
genes	NN	O	O
,	NN	O	O
most	NN	O	O
of	NN	O	O
which	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
globin	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
erythropoietin	NN	O	B-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
Epo-R	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
are	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
relationship	NN	O	O
,	NN	O	O
if	NN	O	O
any	NN	O	O
,	NN	O	O
between	NN	O	O
expression	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
response	NN	O	O
to	NN	O	O
Epo	NN	O	O
and	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
Epo-R	NN	O	B-protein
and	NN	O	O
globin	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
an	NN	O	O
Epo-dependent	NN	O	O
human	NN	O	O
cell	NN	O	O
line	NN	O	O
,	NN	O	O
UT-7	NN	O	B-cell_line
Epo	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
were	NN	O	O
compared	NN	O	O
to	NN	O	O
those	NN	O	O
obtained	NN	O	O
with	NN	O	O
the	NN	O	O
parental	NN	O	B-cell_line
granulocyte-macrophage	NN	O	I-cell_line
colony-stimulating	NN	O	I-cell_line
factor	NN	O	I-cell_line
(	NN	O	I-cell_line
GM-CSF	NN	O	I-cell_line
)	NN	O	I-cell_line
-dependent	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
UT-7	NN	O	B-cell_line
,	NN	O	O
which	NN	O	O
has	NN	O	O
a	NN	O	O
predominantly	NN	O	O
megakaryoblastic	NN	O	O
phenotype	NN	O	O
and	NN	O	O
is	NN	O	O
unable	NN	O	O
to	NN	O	O
proliferate	NN	O	O
continuously	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Epo	NN	O	B-protein
.	NN	O	O

UT-7	NN	O	B-protein
Epo	NN	O	I-protein
and	NN	O	O
UT-7	NN	O	B-cell_line
expressed	NN	O	O
similar	NN	O	O
levels	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
lines	NN	O	O
also	NN	O	O
expressed	NN	O	O
comparable	NN	O	O
levels	NN	O	O
of	NN	O	O
Epo-R	NN	O	B-RNA
mRNA	NN	O	I-RNA
while	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
Epo	NN	O	B-protein
-binding	NN	O	O
sites	NN	O	O
on	NN	O	O
UT-7	NN	O	B-cell_line
Epo	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
one-sixth	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
UT-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
2400	NN	O	O
+/-	NN	O	O
3	NN	O	O
vs.	NN	O	O
13	NN	O	O
,	NN	O	O
800	NN	O	O
+/-	NN	O	O
300	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
could	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
differences	NN	O	O
in	NN	O	O
cell	NN	O	O
surface	NN	O	O
(	NN	O	O
UT-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
20	NN	O	O
%	NN	O	O
smaller	NN	O	O
than	NN	O	O
the	NN	O	O
parental	NN	O	O
UT-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
or	NN	O	O
in	NN	O	O
receptor	NN	O	O
turnover	NN	O	O
.	NN	O	O

By	NN	O	O
Northern	NN	O	O
analysis	NN	O	O
,	NN	O	O
UT-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
expressed	NN	O	O
detectable	NN	O	O
levels	NN	O	O
of	NN	O	O
beta-	NN	O	B-protein
and	NN	O	I-protein
gamma-globin	NN	O	I-protein
but	NN	O	O
not	NN	O	O
alpha-globin	NN	O	B-protein
.	NN	O	O

In	NN	O	O
comparison	NN	O	O
,	NN	O	O
UT-7	NN	O	B-cell_line
Epo	NN	O	I-cell_line
cells	NN	O	I-cell_line
expressed	NN	O	O
alpha-globin	NN	O	O
and	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
gamma-globin	NN	O	B-protein
(	NN	O	O
5-fold	NN	O	O
)	NN	O	O
and	NN	O	O
beta-globin	NN	O	B-protein
(	NN	O	O
from	NN	O	O
barely	NN	O	O
to	NN	O	O
clearly	NN	O	O
detectable	NN	O	O
)	NN	O	O
.	NN	O	O

Globin	NN	O	B-protein
chains	NN	O	I-protein
(	NN	O	O
alpha	NN	O	B-protein
,	NN	O	O
beta	NN	O	B-protein
and	NN	O	O
gamma	NN	O	B-protein
)	NN	O	O
were	NN	O	O
clearly	NN	O	O
detectable	NN	O	O
by	NN	O	O
affinity	NN	O	O
chromatography	NN	O	O
in	NN	O	O
UT-7	NN	O	B-protein
Epo	NN	O	I-protein
but	NN	O	O
not	NN	O	O
in	NN	O	O
UT-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
frequency	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
which	NN	O	O
expressed	NN	O	O
beta-	NN	O	B-DNA
and	NN	O	I-DNA
gamma-	NN	O	I-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
two	NN	O	O
cell	NN	O	O
populations	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
immunofluorescence	NN	O	O
with	NN	O	O
beta-	NN	O	B-protein
and	NN	O	I-protein
gamma-specific	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
gamma-positive	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
their	NN	O	O
fluorescence	NN	O	O
intensity	NN	O	O
were	NN	O	O
higher	NN	O	O
in	NN	O	O
UT-7	NN	O	B-protein
Epo	NN	O	I-protein
than	NN	O	O
in	NN	O	O
UT-7	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
0	NN	O	O
to	NN	O	O
17	NN	O	O
%	NN	O	O
barely	NN	O	O
positive	NN	O	O
cells	NN	O	O
and	NN	O	O
23	NN	O	O
to	NN	O	O
40	NN	O	O
%	NN	O	O
clearly	NN	O	O
positive	NN	O	O
cells	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
globin	NN	O	B-RNA
mRNA	NN	O	I-RNA
observed	NN	O	O
in	NN	O	O
UT-7	NN	O	B-protein
Epo	NN	O	I-protein
is	NN	O	O
due	NN	O	O
to	NN	O	O
both	NN	O	O
an	NN	O	O
increase	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
per	NN	O	O
cell	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
numbers	NN	O	O
of	NN	O	O
cells	NN	O	O
containing	NN	O	O
gamma-globin	NN	O	B-protein
.	NN	O	O

The	NN	O	O
levels	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
Epo-R	NN	O	B-protein
and	NN	O	O
globin	NN	O	B-RNA
mRNA	NN	O	I-RNA
expressed	NN	O	O
were	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
a	NN	O	O
24-hour	NN	O	O
incubation	NN	O	O
of	NN	O	O
either	NN	O	O
cell	NN	O	O
line	NN	O	O
with	NN	O	O
Epo	NN	O	B-protein
,	NN	O	O
GM-CSF	NN	O	B-protein
or	NN	O	O
interleukin-3	NN	O	B-protein
(	NN	O	O
IL-3	NN	O	B-protein
)	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
400	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

The	NN	O	O
Sp1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
binds	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
promoter	NN	O	I-DNA
specifically	NN	O	O
in	NN	O	O
myeloid	NN	O	O
cells	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
myeloid-specific	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

The	NN	O	O
myeloid	NN	O	B-protein
integrin	NN	O	I-protein
CD11b	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
selectively	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
mature	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
,	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Lineage-specific	NN	O	O
expression	NN	O	O
is	NN	O	O
controlled	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
mRNA	NN	O	O
transcription	NN	O	O
.	NN	O	O

Recent	NN	O	O
isolation	NN	O	O
of	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
promoter	NN	O	I-DNA
shows	NN	O	O
that	NN	O	O
92	NN	O	O
base	NN	O	O
pairs	NN	O	O
(	NN	O	O
bp	NN	O	O
)	NN	O	O
of	NN	O	O
5'-flanking	NN	O	B-DNA
DNA	NN	O	I-DNA
are	NN	O	O
sufficient	NN	O	O
to	NN	O	O
direct	NN	O	O
myeloid-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
characterize	NN	O	O
regulatory	NN	O	O
sequences	NN	O	O
important	NN	O	O
for	NN	O	O
promoter	NN	O	O
activity	NN	O	O
,	NN	O	O
we	NN	O	O
performed	NN	O	O
linker	NN	O	O
scanning	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
92-bp	NN	O	B-DNA
CD11b	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
sequence	NN	O	O
at	NN	O	O
bp	NN	O	O
-60	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
CD11b	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
sequence	NN	O	O
binds	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Sp1	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
the	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
bound	NN	O	O
only	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_line
(	NN	O	I-cell_line
U937	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
not	NN	O	O
in	NN	O	O
cervical	NN	O	B-cell_line
carcinoma	NN	O	I-cell_line
(	NN	O	I-cell_line
HeLa	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
macrophage	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
PU.1	NN	O	O
binds	NN	O	O
the	NN	O	O
CD11b	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
Sp1	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
propose	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
myeloid-specific	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
PU.1	NN	O	B-protein
)	NN	O	O
allows	NN	O	O
a	NN	O	O
general	NN	O	O
factor	NN	O	O
(	NN	O	O
Sp1	NN	O	B-protein
)	NN	O	O
to	NN	O	O
bind	NN	O	O
in	NN	O	O
a	NN	O	O
tissue-specific	NN	O	O
fashion	NN	O	O
thereby	NN	O	O
contributing	NN	O	O
to	NN	O	O
the	NN	O	O
myeloid-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
CD11b	NN	O	B-cell_line
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	O
factor	NN	O	O
jun-B	NN	O	O
is	NN	O	O
target	NN	O	O
of	NN	O	O
autoreactive	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
in	NN	O	O
IDDM	NN	O	O
.	NN	O	O

Target	NN	O	O
antigens	NN	O	O
defined	NN	O	O
by	NN	O	O
autoantibodies	NN	O	B-protein
in	NN	O	O
IDDM	NN	O	O
include	NN	O	O
insulin	NN	O	O
,	NN	O	O
a	NN	O	O
putative	NN	O	O
glycolipid	NN	O	O
that	NN	O	O
reacts	NN	O	O
with	NN	O	O
islet	NN	O	B-protein
cell	NN	O	I-protein
antibodies	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
64	NN	O	B-protein
,	NN	O	I-protein
000-M	NN	O	I-protein
(	NN	O	I-protein
r	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
recently	NN	O	O
identified	NN	O	O
as	NN	O	O
glutamic	NN	O	B-protein
acid	NN	O	I-protein
decarboxylase	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
some	NN	O	O
IDDM	NN	O	O
sera	NN	O	O
that	NN	O	O
contain	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	O
glutamic	NN	O	B-protein
acid	NN	O	I-protein
decarboxylase	NN	O	I-protein
also	NN	O	O
coprecipitate	NN	O	O
a	NN	O	O
38	NN	O	B-protein
,	NN	O	I-protein
000-M	NN	O	I-protein
(	NN	O	I-protein
r	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
from	NN	O	O
islets	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
used	NN	O	O
a	NN	O	O
high	NN	O	O
titer	NN	O	O
anti-38	NN	O	O
,	NN	O	O
000-M	NN	O	O
(	NN	O	O
r	NN	O	O
)	NN	O	O
serum	NN	O	O
to	NN	O	O
screen	NN	O	O
bacteriophage	NN	O	B-DNA
lambda	NN	O	I-DNA
cDNA	NN	O	I-DNA
expression	NN	O	O
libraries	NN	O	O
and	NN	O	O
identified	NN	O	O
human	NN	O	B-cell_type
islet	NN	O	I-cell_type
and	NN	O	O
placental	NN	O	B-cell_line
clones	NN	O	I-cell_line
encoding	NN	O	O
jun-B	NN	O	B-protein
,	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
transcription	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
of	NN	O	O
predicted	NN	O	O
38	NN	O	O
,	NN	O	O
000	NN	O	O
M	NN	O	O
(	NN	O	O
r	NN	O	O
)	NN	O	O
.	NN	O	O

Peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T-cells	NN	O	I-cell_type
exhibited	NN	O	O
significant	NN	O	O
proliferation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
a	NN	O	O
recombinant	NN	O	O
fragment	NN	O	O
of	NN	O	O
jun-B	NN	O	B-protein
(	NN	O	O
amino	NN	O	O
acids	NN	O	O
1-180	NN	O	O
)	NN	O	O
in	NN	O	O
12	NN	O	O
of	NN	O	O
17	NN	O	O
(	NN	O	O
71	NN	O	O
%	NN	O	O
)	NN	O	O
recent-onset	NN	O	O
IDDM	NN	O	O
subjects	NN	O	O
,	NN	O	O
8	NN	O	O
of	NN	O	O
16	NN	O	O
(	NN	O	O
50	NN	O	O
%	NN	O	O
)	NN	O	O
ICA-positive	NN	O	O
first-degree	NN	O	O
relatives	NN	O	O
of	NN	O	O
IDDM	NN	O	O
subjects	NN	O	O
who	NN	O	O
were	NN	O	O
at	NN	O	O
risk	NN	O	O
,	NN	O	O
3	NN	O	O
of	NN	O	O
12	NN	O	O
(	NN	O	O
25	NN	O	O
%	NN	O	O
)	NN	O	O
other	NN	O	O
autoimmune	NN	O	O
disease	NN	O	O
subjects	NN	O	O
,	NN	O	O
and	NN	O	O
0	NN	O	O
of	NN	O	O
10	NN	O	O
healthy	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

Proliferation	NN	O	O
to	NN	O	O
tetanus	NN	O	O
toxoid	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
significantly	NN	O	O
between	NN	O	O
the	NN	O	O
groups	NN	O	O
.	NN	O	O

Responses	NN	O	O
to	NN	O	O
jun-B	NN	O	B-protein
were	NN	O	O
not	NN	O	O
related	NN	O	O
to	NN	O	O
age	NN	O	O
,	NN	O	O
sex	NN	O	O
,	NN	O	O
or	NN	O	O
human	NN	O	B-protein
leukocyte	NN	O	I-protein
antigen	NN	O	I-protein
status	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
autoreactive	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
identify	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
antigen	NN	O	I-protein
,	NN	O	O
p38	NN	O	B-protein
jun-B	NN	O	I-protein
,	NN	O	O
in	NN	O	O
IDDM	NN	O	O
and	NN	O	O
appear	NN	O	O
to	NN	O	O
indicate	NN	O	O
subjects	NN	O	O
at	NN	O	O
risk	NN	O	O
for	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
clinical	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stimulation	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
production	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
by	NN	O	O
protein	NN	O	O
phosphatase	NN	O	O
1	NN	O	O
and	NN	O	O
2A	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

Protein	NN	O	B-protein
phosphatases	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	I-protein
2A	NN	O	I-protein
are	NN	O	O
important	NN	O	O
in	NN	O	O
regulating	NN	O	O
cellular	NN	O	O
functions	NN	O	O
by	NN	O	O
controlling	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
state	NN	O	O
of	NN	O	O
their	NN	O	O
substrates	NN	O	O
.	NN	O	O

In	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
inhibitors	NN	O	O
of	NN	O	O
these	NN	O	O
phosphatases	NN	O	B-protein
,	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
and	NN	O	O
calyculin	NN	O	O
A	NN	O	O
,	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
increase	NN	O	O
the	NN	O	O
mRNA	NN	O	O
accumulation	NN	O	O
and	NN	O	O
cytokine	NN	O	O
production	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
and	NN	O	O
interleukin-1	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
increased	NN	O	O
mRNA	NN	O	B-RNA
accumulation	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
primarily	NN	O	O
because	NN	O	O
of	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
transcription	NN	O	O
rate	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-1	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
may	NN	O	O
be	NN	O	O
caused	NN	O	O
by	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
activities	NN	O	O
,	NN	O	O
including	NN	O	O
those	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
by	NN	O	O
these	NN	O	O
protein	NN	O	B-protein
phosphatase	NN	O	I-protein
inhibitors	NN	O	O
.	NN	O	O

Okadaic	NN	O	O
acid	NN	O	O
increased	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
precursor	NN	O	O
and	NN	O	O
mature	NN	O	O
forms	NN	O	O
and	NN	O	O
their	NN	O	O
secretion	NN	O	O
.	NN	O	O

This	NN	O	O
increased	NN	O	O
processing	NN	O	O
and	NN	O	O
secretion	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
IL-1	NN	O	B-RNA
beta	NN	O	I-RNA
convertase	NN	O	I-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
.	NN	O	O

The	NN	O	O
stimulation	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-protein
alpha	NN	O	I-protein
production	NN	O	O
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
was	NN	O	O
more	NN	O	O
modest	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
precursor	NN	O	B-protein
interleukin-1	NN	O	I-protein
alpha	NN	O	I-protein
cytokine	NN	O	I-protein
was	NN	O	O
increased	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
protein	NN	O	O
phosphatase	NN	O	O
1	NN	O	O
and	NN	O	O
2A	NN	O	O
inhibitors	NN	O	O
exert	NN	O	O
multiple	NN	O	O
effects	NN	O	O
on	NN	O	O
cytokine	NN	O	O
production	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
these	NN	O	O
two	NN	O	O
phosphatases	NN	O	B-protein
play	NN	O	O
important	NN	O	O
roles	NN	O	O
in	NN	O	O
regulating	NN	O	O
interleukin-1	NN	O	B-protein
production	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
CD28-responsive	NN	O	I-protein
complex	NN	O	I-protein
contains	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
family	NN	O	I-protein
:	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

Optimal	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
requires	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
signals	NN	O	O
.	NN	O	O

One	NN	O	O
signal	NN	O	O
can	NN	O	O
be	NN	O	O
delivered	NN	O	O
by	NN	O	O
the	NN	O	O
antigen-specific	NN	O	B-protein
T-cell	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
second	NN	O	O
signal	NN	O	O
is	NN	O	O
provided	NN	O	O
by	NN	O	O
the	NN	O	O
costimulatory	NN	O	O
molecule	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
delivered	NN	O	O
by	NN	O	O
the	NN	O	O
antigen-presenting	NN	O	B-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

CD28	NN	O	B-protein
is	NN	O	O
a	NN	O	O
T-cell	NN	O	B-protein
surface	NN	O	I-protein
molecule	NN	O	I-protein
and	NN	O	O
stimulation	NN	O	O
through	NN	O	O
this	NN	O	O
protein	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
delivering	NN	O	O
the	NN	O	O
second	NN	O	O
activation	NN	O	O
signal	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
CD28	NN	O	B-protein
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
involves	NN	O	O
the	NN	O	O
rel	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
--	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
anti-CD28	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
mAb	NN	O	B-protein
)	NN	O	O
results	NN	O	O
in	NN	O	O
augmentation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
c-Rel	NN	O	I-protein
,	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
this	NN	O	O
augmentation	NN	O	O
can	NN	O	O
occur	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
immunosuppressant	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
also	NN	O	O
shown	NN	O	O
in	NN	O	O
this	NN	O	O
report	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
PMA/anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
or	NN	O	O
anti-CD3/anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
,	NN	O	O
c-Rel	NN	O	B-protein
,	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p65	NN	O	B-protein
are	NN	O	O
associated	NN	O	O
with	NN	O	O
CD28-responsive	NN	O	B-DNA
element	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin	NN	O	I-DNA
2	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
functional	NN	O	O
significance	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
involvement	NN	O	O
in	NN	O	O
the	NN	O	O
CD28-responsive	NN	O	B-protein
complex	NN	O	I-protein
is	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
transient	NN	O	O
transfection	NN	O	O
analysis	NN	O	O
,	NN	O	O
where	NN	O	O
cotransfection	NN	O	O
of	NN	O	O
c-Rel	NN	O	B-protein
augments	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
the	NN	O	O
CD28-responsive	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
tal-1	NN	O	B-protein
and	NN	O	O
GATA-binding	NN	O	B-protein
proteins	NN	O	I-protein
during	NN	O	O
human	NN	O	O
hematopoiesis	NN	O	O
.	NN	O	O

Tal-1	NN	O	B-protein
rearrangements	NN	O	O
are	NN	O	O
associated	NN	O	O
with	NN	O	O
nearly	NN	O	O
30	NN	O	O
%	NN	O	O
of	NN	O	O
human	NN	O	O
T	NN	O	O
acute	NN	O	O
lymphoblastic	NN	O	O
leukemia	NN	O	O
.	NN	O	O

Tal-1	NN	O	B-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
putative	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
with	NN	O	O
a	NN	O	O
basic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
domain	NN	O	I-protein
and	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
predominantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
pattern	NN	O	O
of	NN	O	O
tal-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
purified	NN	O	B-cell_type
human	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
and	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
polymerase	NN	O	I-protein
chain	NN	O	O
reaction	NN	O	O
analysis	NN	O	O
.	NN	O	O

Both	NN	O	O
methods	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
tal-1	NN	O	B-protein
gene	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
megakaryocytes	NN	O	O
and	NN	O	O
erythroblasts	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
basophilic	NN	O	B-cell_type
granulocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
tal-1	NN	O	B-DNA
1A	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
contains	NN	O	O
two	NN	O	O
consensus	NN	O	O
GATA-binding	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
active	NN	O	O
mainly	NN	O	O
in	NN	O	O
these	NN	O	O
lineages	NN	O	O
.	NN	O	O

Because	NN	O	O
the	NN	O	O
GATA-1	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
known	NN	O	O
to	NN	O	O
transactivate	NN	O	O
several	NN	O	O
genes	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
erythroid	NN	O	B-cell_type
,	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
mastocytic/basophilic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
studied	NN	O	O
GATA-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
these	NN	O	O
purified	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
GATA-1	NN	O	B-DNA
and	NN	O	I-DNA
tal-1	NN	O	I-DNA
genes	NN	O	I-DNA
are	NN	O	O
coexpressed	NN	O	O
in	NN	O	O
these	NN	O	O
three	NN	O	O
lineages	NN	O	O
.	NN	O	O

Remarkably	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
genes	NN	O	O
is	NN	O	O
downmodulated	NN	O	O
during	NN	O	O
erythroid	NN	O	O
and	NN	O	O
megakaryocytic	NN	O	O
terminal	NN	O	O
maturation	NN	O	O
.	NN	O	O

In	NN	O	O
immature	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
tal-1	NN	O	B-DNA
and	NN	O	I-DNA
GATA-1	NN	O	I-DNA
genes	NN	O	I-DNA
are	NN	O	O
coexpressed	NN	O	O
in	NN	O	O
committed	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
CD34+/CD38	NN	O	B-cell_line
(	NN	O	I-cell_line
2+	NN	O	I-cell_line
)	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
they	NN	O	O
are	NN	O	O
not	NN	O	O
detectable	NN	O	O
in	NN	O	O
the	NN	O	O
most	NN	O	O
primitive	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
CD34	NN	O	B-cell_line
(	NN	O	I-cell_line
2+	NN	O	I-cell_line
)	NN	O	I-cell_line
/CD38-	NN	O	I-cell_line
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
GATA-2	NN	O	B-protein
is	NN	O	O
strongly	NN	O	O
expressed	NN	O	O
in	NN	O	O
both	NN	O	O
most	NN	O	B-cell_type
primitive	NN	O	I-cell_type
and	NN	O	I-cell_type
committed	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
whereas	NN	O	O
GATA-3	NN	O	B-protein
is	NN	O	O
mostly	NN	O	O
detected	NN	O	O
in	NN	O	O
most	NN	O	O
primitive	NN	O	O
ones	NN	O	O
.	NN	O	O

Altogether	NN	O	O
our	NN	O	O
results	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
GATA-1	NN	O	B-protein
modulates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
tal-1	NN	O	B-protein
during	NN	O	O
the	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
erythroid	NN	O	B-cell_type
,	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
,	NN	O	I-cell_type
and	NN	O	I-cell_type
basosophilic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
lytic	NN	O	O
transition	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
is	NN	O	O
imitated	NN	O	O
by	NN	O	O
recombinant	NN	O	B-cell_line
B-cells	NN	O	I-cell_line
.	NN	O	O

Lytic	NN	O	O
transition	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
is	NN	O	O
initiated	NN	O	O
by	NN	O	O
distinct	NN	O	O
immediate	NN	O	O
early	NN	O	O
regulators	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	O
cycle	NN	O	O
,	NN	O	O
in	NN	O	O
synchronization	NN	O	O
to	NN	O	O
temporary	NN	O	O
,	NN	O	O
permissive	NN	O	O
conditions	NN	O	O
during	NN	O	O
host	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

We	NN	O	O
developed	NN	O	O
eukaryotic	NN	O	O
vectors	NN	O	O
suitable	NN	O	O
to	NN	O	O
imitate	NN	O	O
the	NN	O	O
processes	NN	O	O
involved	NN	O	O
in	NN	O	O
lytic	NN	O	O
transition	NN	O	O
in	NN	O	O
cell	NN	O	O
culture	NN	O	O
systems	NN	O	O
.	NN	O	O

Two	NN	O	O
stable	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
established	NN	O	O
:	NN	O	O
R59Z	NN	O	B-cell_line
activator	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
used	NN	O	O
to	NN	O	O
induce	NN	O	O
lytic	NN	O	O
EBV	NN	O	O
expression	NN	O	O
in	NN	O	O
a	NN	O	O
constitutive	NN	O	O
manner	NN	O	O
by	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
the	NN	O	O
BZLF	NN	O	B-protein
1	NN	O	I-protein
trans-activator	NN	O	I-protein
(	NN	O	O
Zta	NN	O	B-protein
)	NN	O	O
.	NN	O	O

R7-57	NN	O	B-cell_line
reporter	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
on	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
signaled	NN	O	O
induced	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
lytic	NN	O	B-DNA
origin	NN	O	I-DNA
of	NN	O	I-DNA
EBV	NN	O	I-DNA
replication	NN	O	I-DNA
(	NN	O	O
ori	NN	O	B-DNA
Lyt	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

Different	NN	O	O
modes	NN	O	O
,	NN	O	O
like	NN	O	O
chemical	NN	O	O
induction	NN	O	O
,	NN	O	O
lytic	NN	O	O
superinfection	NN	O	O
with	NN	O	O
EBV	NN	O	O
and	NN	O	O
single	NN	O	O
gene	NN	O	O
trans-activation	NN	O	O
converted	NN	O	O
the	NN	O	O
recombinant	NN	O	B-DNA
ori	NN	O	I-DNA
Lyt	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
R7-57	NN	O	B-cell_line
reporter	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

BZLF	NN	O	B-protein
1	NN	O	I-protein
,	NN	O	O
transiently	NN	O	O
expressed	NN	O	O
in	NN	O	O
R7-57	NN	O	B-cell_line
reporter	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
was	NN	O	O
the	NN	O	O
only	NN	O	O
EBV	NN	O	B-protein
trans-activator	NN	O	I-protein
found	NN	O	O
,	NN	O	O
sufficient	NN	O	O
in	NN	O	O
inducing	NN	O	O
the	NN	O	O
viral	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
.	NN	O	O

Basing	NN	O	O
on	NN	O	O
these	NN	O	O
experiments	NN	O	O
,	NN	O	O
trans-cellular	NN	O	O
activation	NN	O	O
of	NN	O	O
EBV	NN	O	O
was	NN	O	O
tested	NN	O	O
by	NN	O	O
cocultivation	NN	O	O
of	NN	O	O
BZLF	NN	O	B-cell_type
1-expressing	NN	O	I-cell_type
R59Z	NN	O	I-cell_type
activator	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
R7-57	NN	O	B-cell_line
reporter	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

No	NN	O	O
lytic	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
reporter	NN	O	O
cells	NN	O	O
could	NN	O	O
be	NN	O	O
measured	NN	O	O
,	NN	O	O
neither	NN	O	O
by	NN	O	O
cocultivation	NN	O	O
of	NN	O	O
activator	NN	O	B-cell_line
cells	NN	O	I-cell_line
nor	NN	O	O
by	NN	O	O
coincubation	NN	O	O
of	NN	O	O
BZLF	NN	O	B-protein
1	NN	O	I-protein
-containing	NN	O	O
cell	NN	O	O
lysates	NN	O	O
.	NN	O	O

Latency	NN	O	O
breaking	NN	O	O
activity	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
was	NN	O	O
transferred	NN	O	O
from	NN	O	O
activator	NN	O	B-cell_line
to	NN	O	O
reporter	NN	O	B-cell_line
cells	NN	O	I-cell_line
when	NN	O	O
active	NN	O	O
,	NN	O	O
exogenous	NN	O	O
virus	NN	O	O
was	NN	O	O
added	NN	O	O
.	NN	O	O

The	NN	O	O
cell	NN	O	O
system	NN	O	O
described	NN	O	O
in	NN	O	O
these	NN	O	O
experiments	NN	O	O
provides	NN	O	O
a	NN	O	O
tool	NN	O	O
for	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
EBV	NN	O	O
reactivation	NN	O	O
and	NN	O	O
demonstrates	NN	O	O
the	NN	O	O
potential	NN	O	O
of	NN	O	O
the	NN	O	O
lytic	NN	O	O
regulatory	NN	O	B-DNA
gene	NN	O	I-DNA
BZLF	NN	O	I-DNA
1	NN	O	I-DNA

-DOCSTART-	O

Costimulation	NN	O	O
of	NN	O	O
cAMP	NN	O	O
and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
pathways	NN	O	O
inhibits	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
-dependent	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
persistent	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effects	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
combined	NN	O	O
stimulation	NN	O	O
of	NN	O	O
cAMP	NN	O	O
-and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
-dependent	NN	O	O
pathways	NN	O	O
have	NN	O	O
been	NN	O	O
investigated	NN	O	O
in	NN	O	O
different	NN	O	O
cellular	NN	O	O
systems	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
that	NN	O	O
they	NN	O	O
may	NN	O	O
complement	NN	O	O
each	NN	O	O
other	NN	O	O
in	NN	O	O
activating	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
upon	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
both	NN	O	O
pathways	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
became	NN	O	O
refractory	NN	O	O
to	NN	O	O
activation	NN	O	O
via	NN	O	O
the	NN	O	O
CD3/T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TcR	NN	O	B-protein
)	NN	O	O
complex	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
preincubated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
dibutyryl	NN	O	O
cAMP	NN	O	O
(	NN	O	O
Bt2cAMP	NN	O	O
)	NN	O	O
displayed	NN	O	O
a	NN	O	O
deficient	NN	O	O
proliferative	NN	O	O
ability	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
stimulation	NN	O	O
,	NN	O	O
whereas	NN	O	O
lymphocytes	NN	O	B-cell_type
treated	NN	O	O
individually	NN	O	O
with	NN	O	O
either	NN	O	O
Bt2cAMP	NN	O	O
or	NN	O	O
PMA	NN	O	O
responded	NN	O	O
comparably	NN	O	O
to	NN	O	O
untreated	NN	O	O
samples	NN	O	O
.	NN	O	O

We	NN	O	O
detected	NN	O	O
an	NN	O	O
association	NN	O	O
between	NN	O	O
the	NN	O	O
reduced	NN	O	O
mitogenic	NN	O	O
response	NN	O	O
and	NN	O	O
low	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
chain	NN	O	I-protein
(	NN	O	O
CD25	NN	O	B-protein
)	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
IL-2R	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
mobilization	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
/	NN	O	O
TcR	NN	O	B-protein
-dependent	NN	O	O
signal	NN	O	O
transduction	NN	O	O
was	NN	O	O
impaired	NN	O	O
in	NN	O	O
PMA/Bt2cAMP-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Remarkably	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
these	NN	O	O
samples	NN	O	O
displayed	NN	O	O
a	NN	O	O
persistent	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
protooncogene	NN	O	I-DNA
,	NN	O	O
associated	NN	O	O
to	NN	O	O
an	NN	O	O
increased	NN	O	O
AP-1	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
,	NN	O	O
whereas	NN	O	O
no	NN	O	O
variations	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
or	NN	O	O
NF-kB	NN	O	B-protein
were	NN	O	O
detected	NN	O	O
.	NN	O	O

Neither	NN	O	O
Bt2cAMP	NN	O	O
nor	NN	O	O
PMA	NN	O	O
individually	NN	O	O
mediated	NN	O	O
these	NN	O	O
sustained	NN	O	O
effects	NN	O	O
,	NN	O	O
which	NN	O	O
therefore	NN	O	O
appear	NN	O	O
as	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
the	NN	O	O
interplay	NN	O	O
between	NN	O	O
both	NN	O	O
metabolic	NN	O	O
stimuli	NN	O	O
.	NN	O	O

Altogether	NN	O	O
,	NN	O	O
the	NN	O	O
data	NN	O	O
provide	NN	O	O
the	NN	O	O
evidence	NN	O	O
that	NN	O	O
both	NN	O	O
pathways	NN	O	O
complement	NN	O	O
each	NN	O	O
other	NN	O	O
in	NN	O	O
regulating	NN	O	O
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
,	NN	O	O
conversely	NN	O	O
,	NN	O	O
downregulate	NN	O	O
the	NN	O	O
TcR	NN	O	B-protein
transduction	NN	O	O
mechanisms	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
lymphotoxin	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
stimulated	NN	O	O
by	NN	O	O
HTLV-I	NN	O	B-protein
tax	NN	O	I-protein
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
HTLV-I	NN	O	B-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
tax	NN	O	I-protein
was	NN	O	O
used	NN	O	O
to	NN	O	O
gain	NN	O	O
insight	NN	O	O
into	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
lymphotoxin	NN	O	B-DNA
(	NN	O	I-DNA
LT	NN	O	I-DNA
;	NN	O	I-DNA
TNF-beta	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
induction	NN	O	O
.	NN	O	O

Tax-expressing	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
produce	NN	O	O
LT	NN	O	B-protein
biologic	NN	O	O
activity	NN	O	O
.	NN	O	O

An	NN	O	O
LT	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	I-DNA
LT-293	NN	O	I-DNA
)	NN	O	I-DNA
CAT	NN	O	I-DNA
construct	NN	O	I-DNA
that	NN	O	O
contained	NN	O	O
an	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
active	NN	O	O
in	NN	O	O
the	NN	O	O
LT-producing	NN	O	B-cell_line
C81-66-45	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
contains	NN	O	O
defective	NN	O	O
HTLV-I	NN	O	O
but	NN	O	O
expresses	NN	O	O
tax	NN	O	B-protein
.	NN	O	O

The	NN	O	O
observation	NN	O	O
that	NN	O	O
a	NN	O	O
mutated	NN	O	B-DNA
LT-kappa	NN	O	I-DNA
B	NN	O	I-DNA
construct	NN	O	I-DNA
(	NN	O	O
M1-CAT	NN	O	B-DNA
)	NN	O	O
was	NN	O	O
inactive	NN	O	O
in	NN	O	O
C81-66-45	NN	O	B-cell_line
,	NN	O	O
confirmed	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
LT	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Tax	NN	O	B-protein
was	NN	O	O
transfected	NN	O	O
into	NN	O	O
HTLV-I-negative	NN	O	B-cell_line
human	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
stably	NN	O	O
expressing	NN	O	O
tax	NN	O	B-protein
contained	NN	O	O
elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
that	NN	O	O
directly	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
LT-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Tax	NN	O	B-protein
co-transfected	NN	O	O
with	NN	O	O
reporter	NN	O	B-DNA
constructs	NN	O	I-DNA
into	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
maximally	NN	O	O
activated	NN	O	O
HTLV-I-LTR-CAT	NN	O	B-protein
and	NN	O	O
kappa	NN	O	B-protein
B-fos-CAT	NN	O	I-protein
and	NN	O	O
also	NN	O	O
activated	NN	O	O
LT-293	NN	O	B-DNA
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
.	NN	O	O

In	NN	O	O
JM	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
tax	NN	O	B-protein
induced	NN	O	O
LT-293	NN	O	B-DNA
activity	NN	O	O
by	NN	O	O
two-	NN	O	O
to	NN	O	O
four-fold	NN	O	O
,	NN	O	O
though	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
induction	NN	O	O
of	NN	O	O
M1-CAT	NN	O	B-protein
.	NN	O	O

The	NN	O	O
increase	NN	O	O
in	NN	O	O
LT-293	NN	O	B-DNA
CAT	NN	O	B-protein
activity	NN	O	O
mirrored	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
LT	NN	O	B-protein
biologic	NN	O	O
activity	NN	O	O
seen	NN	O	O
under	NN	O	O
these	NN	O	O
conditions	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
,	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
induction	NN	O	O
of	NN	O	O
LT	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
over	NN	O	O
basal	NN	O	O
levels	NN	O	O
,	NN	O	O
indicate	NN	O	O
that	NN	O	O
HTLV-I	NN	O	B-protein
tax	NN	O	I-protein
causes	NN	O	O
low-level	NN	O	O
activation	NN	O	O
of	NN	O	O
both	NN	O	O
endogenous	NN	O	O
LT	NN	O	B-protein
and	NN	O	O
the	NN	O	O
LT	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
concatenated	NN	O	O
form	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	B-DNA
viral	NN	O	I-DNA
DNA	NN	O	I-DNA
in	NN	O	O
lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
induced	NN	O	O
by	NN	O	O
transfection	NN	O	O
with	NN	O	O
BZLF1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
replicative	NN	O	O
form	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
DNA	NN	O	O
was	NN	O	O
studied	NN	O	O
using	NN	O	O
two	NN	O	O
lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
X50-7	NN	O	O
and	NN	O	O
6F11	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
latently	NN	O	O
infected	NN	O	O
by	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
.	NN	O	O

The	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
of	NN	O	O
EBV	NN	O	O
infection	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
transfection	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
the	NN	O	O
BRLF1/BZLF1	NN	O	B-DNA
coding	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
P3HR-1	NN	O	B-DNA
defective	NN	O	I-DNA
genome	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
combined	NN	O	O
two	NN	O	O
techniques	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
productive	NN	O	O
replicative	NN	O	O
form	NN	O	O
of	NN	O	O
Epstein-Barr	NN	O	B-DNA
viral	NN	O	I-DNA
DNA	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
lytic	NN	O	B-cell_line
cycle-induced	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Restriction	NN	O	O
enzyme	NN	O	O
analysis	NN	O	O
followed	NN	O	O
by	NN	O	O
Southern	NN	O	O
blot	NN	O	O
hybridization	NN	O	O
identified	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
fused	NN	O	O
fragment	NN	O	O
encompassing	NN	O	O
both	NN	O	O
ends	NN	O	O
of	NN	O	O
EBV	NN	O	B-DNA
DNA	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
indicates	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
either	NN	O	O
episomal	NN	O	B-DNA
DNA	NN	O	I-DNA
or	NN	O	O
concatameric	NN	O	B-DNA
linear	NN	O	I-DNA
DNA	NN	O	I-DNA
.	NN	O	O

Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
in	NN	O	O
situ	NN	O	O
lysing	NN	O	O
gels	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
cellular	NN	O	O
content	NN	O	O
of	NN	O	O
linear	NN	O	B-DNA
EBV	NN	O	I-DNA
DNA	NN	O	I-DNA
was	NN	O	O
also	NN	O	O
increased	NN	O	O
significantly	NN	O	O
after	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
circular	NN	O	O
DNA	NN	O	O
remained	NN	O	O
approximately	NN	O	O
constant	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
from	NN	O	O
these	NN	O	O
results	NN	O	O
that	NN	O	O
the	NN	O	O
source	NN	O	O
of	NN	O	O
the	NN	O	O
fused	NN	O	O
fragment	NN	O	O
encompassing	NN	O	O
both	NN	O	O
ends	NN	O	O
of	NN	O	O
EBV	NN	O	B-DNA
DNA	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
concatenated	NN	O	B-DNA
linear	NN	O	I-DNA
EBV	NN	O	I-DNA
DNA	NN	O	I-DNA
molecule	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
that	NN	O	O
such	NN	O	O
a	NN	O	O
concatenated	NN	O	O
molecule	NN	O	O
most	NN	O	O
likely	NN	O	O
represents	NN	O	O
a	NN	O	O
replicative	NN	O	O
form	NN	O	O
of	NN	O	O
EBV	NN	O	B-DNA
DNA	NN	O	I-DNA
in	NN	O	O
productively	NN	O	O
infected	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
thyrotropin	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
autoimmune	NN	O	O
thyroid	NN	O	O
disease	NN	O	O
:	NN	O	O
coregulation	NN	O	O
with	NN	O	O
parameters	NN	O	O
of	NN	O	O
thyroid	NN	O	O
function	NN	O	O
and	NN	O	O
inverse	NN	O	O
relation	NN	O	O
to	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
classes	NN	O	I-protein
I	NN	O	I-protein
and	NN	O	I-protein
II	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
TSH	NN	O	I-DNA
receptor	NN	O	I-DNA
(	NN	O	I-DNA
TSH-R	NN	O	I-DNA
)	NN	O	I-DNA
cDNA	NN	O	I-DNA
probe	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
TSH-R	NN	O	B-protein
transcript	NN	O	O
levels	NN	O	O
in	NN	O	O
13	NN	O	O
human	NN	O	O
thyroid	NN	O	O
fragments	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
;	NN	O	O
7	NN	O	O
Graves	NN	O	O
'	NN	O	O
disease	NN	O	O
,	NN	O	O
2	NN	O	O
Hashimoto	NN	O	O
's	NN	O	O
disease	NN	O	O
,	NN	O	O
3	NN	O	O
endemic	NN	O	O
goiter	NN	O	O
,	NN	O	O
and	NN	O	O
1	NN	O	O
healthy	NN	O	O
thyroid	NN	O	O
gland	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

TSH-R	NN	O	B-protein
expression	NN	O	O
levels	NN	O	O
were	NN	O	O
variable	NN	O	O
,	NN	O	O
but	NN	O	O
displayed	NN	O	O
a	NN	O	O
close	NN	O	O
correlation	NN	O	O
to	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
thyroid	NN	O	B-protein
peroxidase	NN	O	I-protein
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.703	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
0.05	NN	O	O
)	NN	O	O
,	NN	O	O
thyroglobulin	NN	O	B-protein
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.817	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
nuclear	NN	O	B-DNA
oncogene	NN	O	I-DNA
c-fos	NN	O	I-DNA
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.935	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
c-myc	NN	O	B-protein
.	NN	O	O

Overall	NN	O	O
,	NN	O	O
TSH-R	NN	O	B-protein
transcript	NN	O	O
levels	NN	O	O
were	NN	O	O
low	NN	O	O
or	NN	O	O
absent	NN	O	O
in	NN	O	O
those	NN	O	O
thyroids	NN	O	O
in	NN	O	O
which	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-protein
histocompatibility	NN	O	I-protein
complex	NN	O	I-protein
class	NN	O	I-protein
I	NN	O	I-protein
or	NN	O	I-protein
II	NN	O	I-protein
(	NN	O	O
MHC	NN	O	B-protein
I	NN	O	I-protein
or	NN	O	I-protein
II	NN	O	I-protein
)	NN	O	O
was	NN	O	O
high	NN	O	O
,	NN	O	O
thus	NN	O	O
establishing	NN	O	O
an	NN	O	O
inverse	NN	O	O
relation	NN	O	O
(	NN	O	O
MHC	NN	O	B-protein
I	NN	O	I-protein
,	NN	O	O
r	NN	O	O
=	NN	O	O
-0.791	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
;	NN	O	O
MHC	NN	O	B-protein
II	NN	O	I-protein
,	NN	O	O
r	NN	O	O
=	NN	O	O
-0.784	NN	O	O
;	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
showed	NN	O	O
that	NN	O	O
apart	NN	O	O
from	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
thyroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
themselves	NN	O	O
were	NN	O	O
the	NN	O	O
source	NN	O	O
of	NN	O	O
MHC	NN	O	B-protein
II	NN	O	I-protein
transcripts	NN	O	I-protein
.	NN	O	O

gamma-Interferon	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
only	NN	O	O
detectable	NN	O	O
in	NN	O	O
1	NN	O	O
Hashimoto	NN	O	O
's	NN	O	O
goiter	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
next	NN	O	O
to	NN	O	O
lymphocyte	NN	O	O
infiltration	NN	O	O
,	NN	O	O
active	NN	O	O
regulatory	NN	O	O
events	NN	O	O
in	NN	O	O
the	NN	O	O
thyrocyte	NN	O	B-cell_type
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
inverse	NN	O	O
relation	NN	O	O
between	NN	O	O
functional	NN	O	O
parameters	NN	O	O
(	NN	O	O
TSH-R	NN	O	B-protein
,	NN	O	O
thyroid	NN	O	B-protein
peroxidase	NN	O	I-protein
,	NN	O	O
thyroglobulin	NN	O	B-protein
,	NN	O	O
and	NN	O	O
c-fos	NN	O	O
)	NN	O	O
and	NN	O	O
immunological	NN	O	O
markers	NN	O	O
(	NN	O	O
MHC	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
II	NN	O	I-protein
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
antagonism	NN	O	O
between	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
and	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
CD14	NN	O	O
and	NN	O	O
CD23	NN	O	O
expression	NN	O	O
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

1	NN	O	O
,	NN	O	O
25	NN	O	O
alpha-Dihydroxicholecalciferol	NN	O	O
(	NN	O	O
VitD3	NN	O	O
)	NN	O	O
and	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
are	NN	O	O
important	NN	O	O
regulators	NN	O	O
of	NN	O	O
the	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
several	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

This	NN	O	O
paper	NN	O	O
describes	NN	O	O
how	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte-macrophage	NN	O	B-protein
Ag	NN	O	I-protein
,	NN	O	O
CD14	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
low	NN	O	O
affinity	NN	O	B-protein
Fc	NN	O	I-protein
receptor	NN	O	I-protein
for	NN	O	O
IgE	NN	O	B-protein
,	NN	O	O
CD23	NN	O	B-protein
,	NN	O	O
were	NN	O	O
inversely	NN	O	O
regulated	NN	O	O
during	NN	O	O
VitD3	NN	O	O
-and	NN	O	O
RA-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
U-937	NN	O	I-cell_line
monoblasts	NN	O	I-cell_line
.	NN	O	O

PMA	NN	O	O
induced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
CD14	NN	O	B-protein
and	NN	O	O
CD23	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
.	NN	O	O

Exposure	NN	O	O
to	NN	O	O
VitD3	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
the	NN	O	O
de	NN	O	O
novo	NN	O	O
expression	NN	O	O
of	NN	O	O
CD14	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
cycloheximide	NN	O	O
completely	NN	O	O
blocked	NN	O	O
the	NN	O	O
VitD3	NN	O	O
induction	NN	O	O
of	NN	O	O
CD14	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
ongoing	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

While	NN	O	O
inducing	NN	O	O
CD14	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
VitD3	NN	O	O
concomitantly	NN	O	O
suppressed	NN	O	O
the	NN	O	O
basal	NN	O	O
,	NN	O	O
PMA-	NN	O	O
,	NN	O	O
and	NN	O	O
RA-inducible	NN	O	O
CD23	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
induced	NN	O	O
by	NN	O	O
RA	NN	O	O
strongly	NN	O	O
increased	NN	O	O
their	NN	O	O
expression	NN	O	O
of	NN	O	O
CD23	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
,	NN	O	O
whereas	NN	O	O
they	NN	O	O
completely	NN	O	O
lacked	NN	O	O
detectable	NN	O	O
CD14	NN	O	O
cell	NN	O	O
surface	NN	O	O
or	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
VitD3	NN	O	O
-and	NN	O	O
the	NN	O	O
PMA-induced	NN	O	O
CD14	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
inhibited	NN	O	O
as	NN	O	O
a	NN	O	O
temporal	NN	O	O
consequence	NN	O	O
of	NN	O	O
the	NN	O	O
RA-induced	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
there	NN	O	O
exists	NN	O	O
a	NN	O	O
functional	NN	O	O
antagonism	NN	O	O
between	NN	O	O
VitD3	NN	O	O
and	NN	O	O
RA	NN	O	O
that	NN	O	O
may	NN	O	O
have	NN	O	O
important	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
certain	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
through	NN	O	O
their	NN	O	O
inverse	NN	O	O
effects	NN	O	O
on	NN	O	O
CD14	NN	O	O
and	NN	O	O
CD23	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cell-specific	NN	O	O
bifunctional	NN	O	O
role	NN	O	O
of	NN	O	O
Jun	NN	O	B-DNA
oncogene	NN	O	I-DNA
family	NN	O	I-DNA
members	NN	O	I-DNA
on	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

Interaction	NN	O	O
between	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
-	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
-mediated	NN	O	O
signaling	NN	O	O
is	NN	O	O
suggested	NN	O	O
by	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
the	NN	O	O
PKC	NN	O	B-protein
activating	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
to	NN	O	O
inhibit	NN	O	O
GR	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	O
mammary	NN	O	O
tumor	NN	O	O
virus	NN	O	O
(	NN	O	O
MMTV	NN	O	O
)	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
this	NN	O	O
interference	NN	O	O
is	NN	O	O
cell	NN	O	O
specific	NN	O	O
,	NN	O	O
as	NN	O	O
TPA	NN	O	O
augmented	NN	O	O
dexamethasone-induced	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
MMTV	NN	O	B-DNA
LTR	NN	O	I-DNA
in	NN	O	O
several	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
but	NN	O	O
was	NN	O	O
inhibitory	NN	O	O
in	NN	O	O
NIH-3T3	NN	O	B-cell_line
fibroblasts	NN	O	I-cell_line
.	NN	O	O

TPA-GR	NN	O	O
synergism	NN	O	O
was	NN	O	O
determined	NN	O	O
to	NN	O	O
have	NN	O	O
occurred	NN	O	O
at	NN	O	O
the	NN	O	O
GR-responsive	NN	O	B-DNA
element	NN	O	I-DNA
(	NN	O	O
GRE	NN	O	B-DNA
)	NN	O	O
level	NN	O	O
by	NN	O	O
functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
deletion	NN	O	B-DNA
mutants	NN	O	I-DNA
or	NN	O	O
synthetic	NN	O	B-DNA
GRE	NN	O	I-DNA
oligonucleotides	NN	O	I-DNA
driving	NN	O	O
chloramphenicol	NN	O	B-protein
acetyl-transferase	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Synergism	NN	O	O
required	NN	O	O
an	NN	O	O
intact	NN	O	O
GR	NN	O	B-protein
DNA-binding	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
amino-	NN	O	B-protein
or	NN	O	I-protein
carboxyl-terminal	NN	O	I-protein
domains	NN	O	I-protein
were	NN	O	O
dispensable	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
was	NN	O	O
abrogated	NN	O	O
by	NN	O	O
the	NN	O	O
PKC	NN	O	B-protein
inhibitor	NN	O	O
staurosporine	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
PKC	NN	O	B-protein
.	NN	O	O

Increased	NN	O	O
c-jun	NN	O	B-protein
,	NN	O	O
jun-B	NN	O	B-protein
,	NN	O	O
and	NN	O	O
jun-D	NN	O	B-protein
expression	NN	O	O
above	NN	O	O
basal	NN	O	O
levels	NN	O	O
and	NN	O	O
increased	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
AP-1/TPA	NN	O	B-DNA
responsive	NN	O	I-DNA
elements	NN	O	I-DNA
fused	NN	O	O
to	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyl-transferase	NN	O	I-DNA
vectors	NN	O	I-DNA
were	NN	O	O
observed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
TPA	NN	O	O
alone	NN	O	O
or	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
Jun	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
cooperate	NN	O	O
with	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
has	NN	O	O
been	NN	O	O
investigated	NN	O	O
after	NN	O	O
transfection	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	I-DNA
jun-B	NN	O	I-DNA
,	NN	O	I-DNA
or	NN	O	I-DNA
jun-D	NN	O	I-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
augmented	NN	O	O
GR	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
from	NN	O	O
either	NN	O	O
MMTV	NN	O	B-DNA
LTR	NN	O	I-DNA
or	NN	O	O
GRE	NN	O	B-DNA
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
c-jun	NN	O	B-protein
and	NN	O	O
jun-B	NN	O	B-protein
transfection	NN	O	O
blunted	NN	O	O
GR	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
c-fos	NN	O	B-protein
had	NN	O	O
a	NN	O	O
negative	NN	O	O
influence	NN	O	O
on	NN	O	O
GR	NN	O	B-protein
function	NN	O	O
and	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
cell-specific	NN	O	O
synergistic	NN	O	O
or	NN	O	O
antagonistic	NN	O	O
activity	NN	O	O
of	NN	O	O
Jun	NN	O	B-protein
with	NN	O	O
respect	NN	O	O
to	NN	O	O
GR	NN	O	B-protein
;	NN	O	O
high	NN	O	O
basal	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
AP-1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
overexpression	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
c-fos	NN	O	I-protein
has	NN	O	O
an	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
GR	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
from	NN	O	O
GRE	NN	O	B-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
Jun	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
bifunctional	NN	O	O
role	NN	O	O
on	NN	O	O
GR	NN	O	B-protein
-dependent	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
GRE	NN	O	B-DNA
,	NN	O	O
selecting	NN	O	O
either	NN	O	O
synergistic	NN	O	O
or	NN	O	O
antagonistic	NN	O	O
activity	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
cell-specific	NN	O	O
microenvironment	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
regard	NN	O	O
,	NN	O	O
intracellular	NN	O	O
levels	NN	O	O
of	NN	O	O
c-fos	NN	O	B-protein
appear	NN	O	O
to	NN	O	O
be	NN	O	O
influential	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cell	NN	O	O
type-	NN	O	O
and	NN	O	O
stage-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CD20/B1	NN	O	B-protein
antigen	NN	O	I-protein
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
diverged	NN	O	O
octamer	NN	O	O
DNA	NN	O	O
motif	NN	O	O
present	NN	O	O
in	NN	O	O
its	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
CD20	NN	O	B-DNA
(	NN	O	I-DNA
B1	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
protein	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	O
B	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Studies	NN	O	O
with	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
deletion	NN	O	I-DNA
CD20	NN	O	I-DNA
promoter-CAT	NN	O	I-DNA
constructs	NN	O	I-DNA
have	NN	O	O
previously	NN	O	O
revealed	NN	O	O
two	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
between	NN	O	O
bases	NN	O	O
-186	NN	O	O
and	NN	O	O
-280	NN	O	O
and	NN	O	O
between	NN	O	O
bases	NN	O	O
-280	NN	O	O
and	NN	O	O
-454	NN	O	O
which	NN	O	O
contained	NN	O	O
positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
sequence	NN	O	B-DNA
element	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
the	NN	O	O
most	NN	O	O
proximal	NN	O	O
region	NN	O	O
located	NN	O	O
between	NN	O	O
bases	NN	O	O
-214	NN	O	O
and	NN	O	O
-201	NN	O	O
,	NN	O	O
TTCTTCTAATTAA	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
high	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
CD20	NN	O	B-protein
in	NN	O	O
mature	NN	O	O
B	NN	O	O
cells	NN	O	O
and	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
CD20	NN	O	B-protein
in	NN	O	O
pre-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
sequence	NN	O	B-DNA
element	NN	O	I-DNA
was	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
the	NN	O	O
BAT	NN	O	B-DNA
box	NN	O	I-DNA
and	NN	O	O
its	NN	O	O
deletion	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
CD20	NN	O	O
promoter-CAT	NN	O	O
construct	NN	O	O
in	NN	O	O
B	NN	O	O
cells	NN	O	O
.	NN	O	O

Mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
with	NN	O	O
various	NN	O	O
mutant	NN	O	O
probes	NN	O	O
and	NN	O	O
B	NN	O	O
cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
core	NN	O	O
sequence	NN	O	O
TAAT	NN	O	O
was	NN	O	O
essential	NN	O	O
for	NN	O	O
binding	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
.	NN	O	O

Cross	NN	O	O
competition	NN	O	O
experiments	NN	O	O
with	NN	O	O
an	NN	O	O
octamer	NN	O	O
sequence	NN	O	O
from	NN	O	O
the	NN	O	O
Ig	NN	O	O
heavy	NN	O	O
chain	NN	O	O
promoter	NN	O	O
,	NN	O	O
the	NN	O	O
BAT	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
TA-rich	NN	O	O
sequence	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
CD21	NN	O	B-DNA
promoter	NN	O	I-DNA
revealed	NN	O	O
that	NN	O	O
all	NN	O	O
three	NN	O	O
sequences	NN	O	O
bound	NN	O	O
the	NN	O	O
same	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
BAT	NN	O	B-protein
box	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
were	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
.	NN	O	O

Southwestern	NN	O	O
blotting	NN	O	O
and	NN	O	O
UV	NN	O	O
cross-linking	NN	O	O
studies	NN	O	O
confirmed	NN	O	O
that	NN	O	O
the	NN	O	O
BAT	NN	O	B-protein
box	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
were	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
affinity	NN	O	O
of	NN	O	O
the	NN	O	O
BAT	NN	O	B-protein
box	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
for	NN	O	O
the	NN	O	O
BAT	NN	O	B-DNA
box	NN	O	I-DNA
was	NN	O	O
approximately	NN	O	O
25-fold	NN	O	O
less	NN	O	O
than	NN	O	O
for	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
sequence	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
BAT	NN	O	B-protein
box	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
dissociated	NN	O	O
from	NN	O	O
the	NN	O	O
BAT	NN	O	B-DNA
box	NN	O	I-DNA
10-fold	NN	O	O
more	NN	O	O
rapidly	NN	O	O
than	NN	O	O
from	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

Despite	NN	O	O
this	NN	O	O
lower	NN	O	O
affinity	NN	O	O
,	NN	O	O
a	NN	O	O
trimer	NN	O	O
of	NN	O	O
the	NN	O	O
BAT	NN	O	B-DNA
box	NN	O	I-DNA
sequence	NN	O	I-DNA
was	NN	O	O
as	NN	O	O
efficiently	NN	O	O
transactivated	NN	O	O
by	NN	O	O
an	NN	O	O
Oct-2	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
as	NN	O	O
was	NN	O	O
a	NN	O	O
trimer	NN	O	O
of	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
sequence	NN	O	I-DNA
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
BAT	NN	O	B-DNA
box	NN	O	I-DNA
and	NN	O	O
Oct-2	NN	O	B-protein
were	NN	O	O
also	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
CD20	NN	O	B-protein
in	NN	O	O
the	NN	O	O
pre-B	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
PB-697	NN	O	B-cell_line
,	NN	O	O
via	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
CD20	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
temporally	NN	O	O
associated	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
Oct-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
a	NN	O	O
BAT	NN	O	B-DNA
box-deleted	NN	O	I-DNA
CD20-CAT	NN	O	I-DNA
construct	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
wild	NN	O	O
type	NN	O	O
,	NN	O	O
was	NN	O	O
poorly	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
.	NN	O	O

Together	NN	O	O
these	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
BAT	NN	O	B-protein
box	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
B	NN	O	O
cell	NN	O	O
specific	NN	O	O
expression	NN	O	O
of	NN	O	O
CD20	NN	O	B-protein
and	NN	O	O
perhaps	NN	O	O
CD21	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-cell_type
CD4	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
specifically	NN	O	O
recognize	NN	O	O
a	NN	O	O
peptide	NN	O	O
representing	NN	O	O
the	NN	O	O
fusion	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
hybrid	NN	O	B-protein
protein	NN	O	I-protein
pml/RAR	NN	O	B-protein
alpha	NN	O	I-protein
present	NN	O	O
in	NN	O	O
acute	NN	O	B-cell_type
promyelocytic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Fusion	NN	O	O
proteins	NN	O	O
present	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
frequently	NN	O	O
contain	NN	O	O
a	NN	O	O
new	NN	O	O
amino	NN	O	O
acid	NN	O	O
at	NN	O	O
the	NN	O	O
fusion	NN	O	B-protein
point	NN	O	I-protein
.	NN	O	O

We	NN	O	O
tested	NN	O	O
whether	NN	O	O
a	NN	O	O
peptide	NN	O	O
(	NN	O	O
BCR1/25	NN	O	O
)	NN	O	O
encompassing	NN	O	O
the	NN	O	O
fusion	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
hybrid	NN	O	O
molecule	NN	O	O
pml/RAR	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
selectively	NN	O	O
expressed	NN	O	O
by	NN	O	O
acute	NN	O	B-cell_type
promyelocytic	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
(	NN	O	I-cell_type
APL	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
can	NN	O	O
be	NN	O	O
recognized	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

CD4+	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
at	NN	O	O
both	NN	O	O
polyclonal	NN	O	O
and	NN	O	O
clonal	NN	O	O
level	NN	O	O
,	NN	O	O
recognized	NN	O	O
peptide	NN	O	O
BCR1/25	NN	O	O
in	NN	O	O
an	NN	O	O
HLA-DR	NN	O	B-protein
--	NN	O	O
restricted	NN	O	O
fashion	NN	O	O
on	NN	O	O
presentation	NN	O	O
by	NN	O	O
autologous	NN	O	B-cell_type
antigen-presenting	NN	O	I-cell_type
cell	NN	O	I-cell_type
(	NN	O	O
APC	NN	O	B-cell_type
)	NN	O	O
or	NN	O	O
by	NN	O	O
APC	NN	O	B-cell_type
expressing	NN	O	O
the	NN	O	O
HLA-DR11	NN	O	B-protein
restricting	NN	O	I-protein
molecule	NN	O	I-protein
.	NN	O	O

Control	NN	O	O
peptides	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
normal	NN	O	O
pml	NN	O	B-protein
and	NN	O	O
RAR	NN	O	B-protein
alpha	NN	O	I-protein
proteins	NN	O	I-protein
were	NN	O	O
not	NN	O	O
recognized	NN	O	O
.	NN	O	O

One	NN	O	O
clone	NN	O	O
(	NN	O	O
DEG5	NN	O	B-cell_line
)	NN	O	O
also	NN	O	O
exerted	NN	O	O
a	NN	O	O
high	NN	O	O
and	NN	O	O
specific	NN	O	O
cytotoxicity	NN	O	O
against	NN	O	O
autologous	NN	O	B-cell_line
cells	NN	O	I-cell_line
pulsed	NN	O	O
with	NN	O	O
BCR1/25	NN	O	O
.	NN	O	O

The	NN	O	O
autologous	NN	O	B-cell_line
DE	NN	O	I-cell_line
LCL	NN	O	I-cell_line
containing	NN	O	O
a	NN	O	O
transduced	NN	O	O
pml/RAR	NN	O	B-DNA
alpha	NN	O	I-DNA
fusion	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
expressing	NN	O	O
a	NN	O	O
bcr1	NN	O	O
type	NN	O	O
of	NN	O	O
the	NN	O	O
pml/RAR	NN	O	B-protein
alpha	NN	O	I-protein
hybrid	NN	O	O
protein	NN	O	O
induced	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
DE	NN	O	B-cell_line
anti-BCR1/25	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
clones	NN	O	I-cell_line
.	NN	O	O

It	NN	O	O
is	NN	O	O
concluded	NN	O	O
that	NN	O	O
the	NN	O	O
bcr1	NN	O	B-protein
type-pml/RAR	NN	O	I-protein
alpha	NN	O	I-protein
fusion	NN	O	I-protein
protein	NN	O	I-protein
of	NN	O	O
APL	NN	O	B-cell_line
contains	NN	O	O
an	NN	O	O
antigenic	NN	O	O
site	NN	O	O
,	NN	O	O
absent	NN	O	O
from	NN	O	O
the	NN	O	O
normal	NN	O	O
parent	NN	O	O
molecules	NN	O	O
and	NN	O	O
recognized	NN	O	O
by	NN	O	O
human	NN	O	B-cell_line
CD4+	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Egr-1	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
and	NN	O	O
restricts	NN	O	O
differentiation	NN	O	O
along	NN	O	O
the	NN	O	O
macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
isolated	NN	O	O
cDNA	NN	O	B-DNA
clones	NN	O	I-DNA
of	NN	O	O
myeloid	NN	O	B-DNA
differentiation	NN	O	I-DNA
primary	NN	O	I-DNA
response	NN	O	I-DNA
(	NN	O	I-DNA
MyD	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
activated	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
following	NN	O	O
induction	NN	O	O
for	NN	O	O
differentiation	NN	O	O
along	NN	O	O
either	NN	O	O
the	NN	O	O
macrophage	NN	O	B-cell_type
or	NN	O	O
granulocyte	NN	O	B-cell_type
lineage	NN	O	I-cell_type
in	NN	O	O
human	NN	O	O
myeloblastic	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

One	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
of	NN	O	O
a	NN	O	O
primary	NN	O	O
response	NN	O	O
gene	NN	O	O
,	NN	O	O
expressed	NN	O	O
upon	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
,	NN	O	O
encoded	NN	O	O
for	NN	O	O
Egr-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Egr-1	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
observed	NN	O	O
to	NN	O	O
be	NN	O	O
transcriptionally	NN	O	O
silent	NN	O	O
in	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
active	NN	O	O
in	NN	O	O
U-937	NN	O	B-cell_line
and	NN	O	O
M1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
latter	NN	O	O
two	NN	O	O
being	NN	O	O
predetermined	NN	O	O
for	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Egr-1	NN	O	B-protein
antisense	NN	O	O
oligomers	NN	O	O
in	NN	O	O
the	NN	O	O
culture	NN	O	O
media	NN	O	O
blocked	NN	O	O
macrophage	NN	O	O
differentiation	NN	O	O
in	NN	O	O
both	NN	O	O
myeloid	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
normal	NN	O	O
myeloblasts	NN	O	B-cell_type
.	NN	O	O

HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
constitutively	NN	O	O
expressing	NN	O	O
an	NN	O	O
Egr-1	NN	O	B-DNA
transgene	NN	O	I-DNA
(	NN	O	O
HL-60Egr-1	NN	O	B-DNA
)	NN	O	O
could	NN	O	O
be	NN	O	O
induced	NN	O	O
for	NN	O	O
macrophage	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
granulocyte	NN	O	B-cell_type
,	NN	O	O
differentiation	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
indicate	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
Egr-1	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
and	NN	O	O
restricts	NN	O	O
differentiation	NN	O	O
of	NN	O	O
myeloblasts	NN	O	B-cell_type
along	NN	O	O
the	NN	O	O
macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
2	NN	O	I-protein
interacts	NN	O	O
with	NN	O	O
an	NN	O	O
EBNA2	NN	O	B-DNA
responsive	NN	O	I-DNA
cis-element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
terminal	NN	O	B-DNA
protein	NN	O	I-DNA
1	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
protein	NN	O	I-protein
EBNA2	NN	O	I-protein
acts	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
of	NN	O	O
cellular	NN	O	B-DNA
and	NN	O	I-DNA
viral	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
plays	NN	O	O
a	NN	O	O
crucial	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
immortalization	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
primary	NN	O	I-cell_type
B-cells	NN	O	I-cell_type
by	NN	O	O
EBV	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
previously	NN	O	O
that	NN	O	O
EBNA2	NN	O	B-protein
transactivates	NN	O	O
the	NN	O	O
promoters	NN	O	O
of	NN	O	O
the	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
antigens	NN	O	I-protein
LMP	NN	O	B-protein
,	NN	O	O
TP1	NN	O	B-protein
and	NN	O	O
TP2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
TP1	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
chosen	NN	O	O
as	NN	O	O
a	NN	O	O
model	NN	O	O
system	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
of	NN	O	O
EBNA2	NN	O	B-protein
mediated	NN	O	O
transactivation	NN	O	O
.	NN	O	O

To	NN	O	O
identify	NN	O	O
an	NN	O	O
EBNA2	NN	O	B-DNA
dependent	NN	O	I-DNA
cis-acting	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
various	NN	O	O
TP1	NN	O	B-DNA
promoter-reporter	NN	O	I-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
were	NN	O	O
transfected	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
and	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
EBNA2	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
into	NN	O	O
the	NN	O	O
established	NN	O	O
B-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
BL41-P3HR1	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
delineate	NN	O	O
an	NN	O	O
81	NN	O	B-DNA
bp	NN	O	I-DNA
EBNA2	NN	O	I-DNA
responsive	NN	O	I-DNA
region	NN	O	I-DNA
between	NN	O	O
-258	NN	O	O
and	NN	O	O
-177	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
TP1	NN	O	B-DNA
RNA	NN	O	I-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
element	NN	O	O
worked	NN	O	O
in	NN	O	O
either	NN	O	O
orientation	NN	O	O
and	NN	O	O
could	NN	O	O
mediate	NN	O	O
EBNA2	NN	O	B-protein
dependent	NN	O	O
transactivation	NN	O	O
on	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
revealed	NN	O	O
three	NN	O	O
specific	NN	O	O
protein-DNA	NN	O	B-protein
complexes	NN	O	I-protein
formed	NN	O	O
with	NN	O	O
sequences	NN	O	O
of	NN	O	O
the	NN	O	O
EBNA2	NN	O	B-DNA
responsive	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Two	NN	O	O
of	NN	O	O
these	NN	O	O
were	NN	O	O
not	NN	O	O
cell	NN	O	O
type	NN	O	O
specific	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
third	NN	O	O
was	NN	O	O
detected	NN	O	O
only	NN	O	O
in	NN	O	O
EBNA2	NN	O	B-protein
positive	NN	O	O
cell	NN	O	O
extracts	NN	O	O
.	NN	O	O

Gel-shift	NN	O	O
analysis	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
EBNA2	NN	O	B-protein
specific	NN	O	O
monoclonal	NN	O	O
antibodies	NN	O	O
revealed	NN	O	O
that	NN	O	O
EBNA2	NN	O	B-protein
is	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
third	NN	O	O
complex	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
these	NN	O	O
experiments	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
EBNA2	NN	O	B-protein
interacts	NN	O	O
with	NN	O	O
an	NN	O	O
EBNA2	NN	O	B-DNA
responsive	NN	O	I-DNA
cis-element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
TP1	NN	O	B-DNA
promoter	NN	O	I-DNA

-DOCSTART-	O

Minimally	NN	O	O
modified	NN	O	O
low	NN	O	O
density	NN	O	O
lipoprotein	NN	O	B-protein
-induced	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
cyclic	NN	O	O
adenosine	NN	O	O
monophosphate	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
minimally	NN	O	B-cell_line
oxidized	NN	O	I-cell_line
LDL	NN	O	I-cell_line
(	NN	O	I-cell_line
MM-LDL	NN	O	I-cell_line
)	NN	O	I-cell_line
activated	NN	O	I-cell_line
endothelial	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
increase	NN	O	O
their	NN	O	O
interaction	NN	O	O
with	NN	O	O
monocytes	NN	O	B-cell_type
but	NN	O	O
not	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
inducing	NN	O	O
monocyte	NN	O	O
but	NN	O	O
not	NN	O	O
neutrophil	NN	O	O
binding	NN	O	O
and	NN	O	O
synthesis	NN	O	O
of	NN	O	O
monocyte	NN	O	B-protein
chemotactic	NN	O	I-protein
protein-1	NN	O	I-protein
and	NN	O	O
monocyte	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
M-CSF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
studies	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
by	NN	O	O
which	NN	O	O
this	NN	O	O
monocyte-specific	NN	O	O
response	NN	O	O
is	NN	O	O
induced	NN	O	O
.	NN	O	O

Both	NN	O	O
induction	NN	O	O
of	NN	O	O
monocyte	NN	O	O
binding	NN	O	O
and	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
for	NN	O	O
M-CSF	NN	O	B-protein
by	NN	O	O
MM-LDL	NN	O	B-protein
were	NN	O	O
not	NN	O	O
inhibited	NN	O	O
in	NN	O	O
protein	NN	O	O
kinase	NN	O	O
C-depleted	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
number	NN	O	O
of	NN	O	O
our	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
cAMP	NN	O	O
is	NN	O	O
the	NN	O	O
second	NN	O	O
messenger	NN	O	O
for	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
MM-LDL	NN	O	B-protein
cited	NN	O	O
above	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
MM-LDL	NN	O	B-protein
caused	NN	O	O
a	NN	O	O
173	NN	O	O
%	NN	O	O
increase	NN	O	O
in	NN	O	O
intracellular	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
.	NN	O	O

Agents	NN	O	O
which	NN	O	O
increased	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
,	NN	O	O
including	NN	O	O
cholera	NN	O	O
toxin	NN	O	O
,	NN	O	O
pertussis	NN	O	O
toxin	NN	O	O
,	NN	O	O
dibutyryl	NN	O	O
cAMP	NN	O	O
,	NN	O	O
and	NN	O	O
isoproterenol	NN	O	O
mimicked	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
MM-LDL	NN	O	B-protein
.	NN	O	O

Agents	NN	O	O
which	NN	O	O
elevated	NN	O	O
cAMP	NN	O	O
were	NN	O	O
also	NN	O	O
shown	NN	O	O
to	NN	O	O
activate	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
this	NN	O	O
transcription	NN	O	O
factor	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
monocyte-endothelial	NN	O	O
interactions	NN	O	O
.	NN	O	O

Although	NN	O	O
endothelial	NN	O	B-RNA
leukocyte	NN	O	I-RNA
adhesion	NN	O	I-RNA
molecule	NN	O	I-RNA
(	NN	O	I-RNA
ELAM	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
synthesis	NN	O	O
can	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
ELAM	NN	O	B-protein
was	NN	O	O
not	NN	O	O
expressed	NN	O	O
and	NN	O	O
ELAM	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
only	NN	O	O
slightly	NN	O	O
elevated	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
MM-LDL	NN	O	B-protein
.	NN	O	O

We	NN	O	O
present	NN	O	O
evidence	NN	O	O
that	NN	O	O
induction	NN	O	O
of	NN	O	O
neutrophil	NN	O	B-cell_type
binding	NN	O	O
by	NN	O	O
LPS	NN	O	O
is	NN	O	O
actually	NN	O	O
suppressed	NN	O	O
by	NN	O	O
agents	NN	O	O
that	NN	O	O
elevated	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-protein
T	NN	O	I-protein
cell	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
GATA-3	NN	O	B-protein
stimulates	NN	O	O
HIV-1	NN	O	O
expression	NN	O	O
.	NN	O	O

A	NN	O	O
family	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activating	NN	O	O
proteins	NN	O	O
,	NN	O	O
the	NN	O	O
GATA	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
a	NN	O	O
consensus	NN	O	O
motif	NN	O	O
through	NN	O	O
a	NN	O	O
highly	NN	O	B-protein
conserved	NN	O	I-protein
C4	NN	O	I-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

One	NN	O	O
member	NN	O	O
of	NN	O	O
this	NN	O	O
multigene	NN	O	O
family	NN	O	O
,	NN	O	O
GATA-3	NN	O	B-protein
,	NN	O	O
is	NN	O	O
most	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
cellular	NN	O	O
target	NN	O	O
for	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
infection	NN	O	O
and	NN	O	O
replication	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
footprinting	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
six	NN	O	O
hGATA-3	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
U3	NN	O	B-DNA
region	NN	O	I-DNA
(	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
regulatory	NN	O	I-DNA
domain	NN	O	I-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

Cotransfection	NN	O	O
of	NN	O	O
an	NN	O	O
hGATA-3	NN	O	B-DNA
expression	NN	O	I-DNA
plasmid	NN	O	I-DNA
with	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
plasmid	NN	O	I-DNA
whose	NN	O	O
transcription	NN	O	O
is	NN	O	O
directed	NN	O	O
by	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
6-	NN	O	O
to	NN	O	O
10-fold	NN	O	O
stimulation	NN	O	O
of	NN	O	O
LTR	NN	O	B-DNA
-mediated	NN	O	O
transcription	NN	O	O
,	NN	O	O
whereas	NN	O	O
site	NN	O	O
specific	NN	O	O
mutation	NN	O	O
of	NN	O	O
these	NN	O	O
GATA	NN	O	B-DNA
sites	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
virtual	NN	O	O
abrogation	NN	O	O
of	NN	O	O
the	NN	O	O
activation	NN	O	O
by	NN	O	O
hGATA-3	NN	O	B-protein
.	NN	O	O

Further	NN	O	O
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
hGATA-3	NN	O	B-DNA
transcriptional	NN	O	I-DNA
activation	NN	O	I-DNA
domain	NN	O	I-DNA
abolished	NN	O	O
GATA-dependent	NN	O	O
HIV-1	NN	O	O
trans-activation	NN	O	O
,	NN	O	O
showing	NN	O	O
that	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
viral	NN	O	O
transcription	NN	O	O
observed	NN	O	O
is	NN	O	O
a	NN	O	O
direct	NN	O	O
effect	NN	O	O
of	NN	O	O
cotransfected	NN	O	B-protein
hGATA-3	NN	O	I-protein
.	NN	O	O

Introduction	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
plasmids	NN	O	I-DNA
in	NN	O	O
which	NN	O	O
the	NN	O	O
GATA	NN	O	B-DNA
sites	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
mutated	NN	O	O
into	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
also	NN	O	O
caused	NN	O	O
a	NN	O	O
significant	NN	O	O
reduction	NN	O	O
in	NN	O	O
LTR	NN	O	B-DNA
-mediated	NN	O	O
transcription	NN	O	O
at	NN	O	O
both	NN	O	O
the	NN	O	O
basal	NN	O	O
level	NN	O	O
and	NN	O	O
in	NN	O	O
(	NN	O	O
PHA-	NN	O	O
plus	NN	O	O
PMA-	NN	O	O
)	NN	O	O
stimulated	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
its	NN	O	O
normal	NN	O	O
role	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
gene	NN	O	O
regulation	NN	O	O
,	NN	O	O
hGATA-3	NN	O	B-protein
may	NN	O	O
also	NN	O	O
play	NN	O	O
a	NN	O	O
significant	NN	O	O
role	NN	O	O
in	NN	O	O
HIV-1	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cytokine	NN	O	O
modulation	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
.	NN	O	O

Cytokines	NN	O	B-protein
,	NN	O	O
the	NN	O	O
peptide	NN	O	B-protein
hormones	NN	O	I-protein
which	NN	O	O
control	NN	O	O
the	NN	O	O
homeostasis	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
and	NN	O	O
also	NN	O	O
play	NN	O	O
a	NN	O	O
fundamental	NN	O	O
role	NN	O	O
in	NN	O	O
inflammatory	NN	O	O
and	NN	O	O
immune	NN	O	O
mediated	NN	O	O
reactions	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
involved	NN	O	O
at	NN	O	O
multiple	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
the	NN	O	O
acquired	NN	O	O
immune	NN	O	O
deficiency	NN	O	O
syndrome	NN	O	O
(	NN	O	O
AIDS	NN	O	O
)	NN	O	O
.	NN	O	O

Infection	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
induce	NN	O	O
production	NN	O	O
of	NN	O	O
several	NN	O	O
cytokines	NN	O	B-protein
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
several	NN	O	O
cytokines	NN	O	B-protein
modulate	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
in	NN	O	O
infected	NN	O	O
cells	NN	O	O
of	NN	O	O
both	NN	O	O
T	NN	O	B-cell_type
lymphocytic	NN	O	I-cell_type
and	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
phagocytic	NN	O	I-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

Activated	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
particularly	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
which	NN	O	O
are	NN	O	O
in	NN	O	O
a	NN	O	O
state	NN	O	O
of	NN	O	O
chronic	NN	O	O
activation	NN	O	O
in	NN	O	O
HIV	NN	O	O
infected	NN	O	O
individuals	NN	O	O
,	NN	O	O
release	NN	O	O
HIV-inductive	NN	O	B-protein
cytokines	NN	O	I-protein
and	NN	O	O
thus	NN	O	O
play	NN	O	O
a	NN	O	O
potentially	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
HIV	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
and	NN	O	O
functional	NN	O	O
characterization	NN	O	O
of	NN	O	O
early	NN	O	B-protein
B-cell	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
a	NN	O	O
regulator	NN	O	O
of	NN	O	O
lymphocyte-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Early	NN	O	B-protein
B-cell	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
EBF	NN	O	B-protein
)	NN	O	O
was	NN	O	O
identified	NN	O	O
previously	NN	O	O
as	NN	O	O
a	NN	O	O
tissue-specific	NN	O	O
and	NN	O	O
differentiation	NN	O	O
stage-specific	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
participates	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
pre-B	NN	O	B-DNA
and	NN	O	I-DNA
B	NN	O	I-DNA
lymphocyte-specific	NN	O	I-DNA
mb-1	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Partial	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequences	NN	O	O
obtained	NN	O	O
from	NN	O	O
purified	NN	O	O
EBF	NN	O	B-protein
were	NN	O	O
used	NN	O	O
to	NN	O	O
isolate	NN	O	O
cDNA	NN	O	B-DNA
clones	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
by	NN	O	O
multiple	NN	O	O
criteria	NN	O	O
encode	NN	O	O
EBF	NN	O	B-protein
.	NN	O	O

The	NN	O	O
recombinant	NN	O	B-protein
polypeptide	NN	O	I-protein
formed	NN	O	O
sequence-specific	NN	O	O
complexes	NN	O	O
with	NN	O	O
the	NN	O	O
EBF-binding	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
mb-1	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
cDNA	NN	O	B-DNA
hybridized	NN	O	O
to	NN	O	O
multiple	NN	O	O
transcripts	NN	O	O
in	NN	O	O
pre-B	NN	O	B-cell_line
and	NN	O	I-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
transcripts	NN	O	O
were	NN	O	O
not	NN	O	O
detected	NN	O	O
at	NN	O	O
significant	NN	O	O
levels	NN	O	O
in	NN	O	O
plasmacytoma	NN	O	B-cell_line
,	NN	O	O
T-cell	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
nonlymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
recombinant	NN	O	O
EBF	NN	O	B-protein
in	NN	O	O
transfected	NN	O	B-cell_line
nonlymphoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
strongly	NN	O	O
activated	NN	O	O
transcription	NN	O	O
from	NN	O	O
reporter	NN	O	O
plasmids	NN	O	O
containing	NN	O	O
functional	NN	O	O
EBF-binding	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
deletion	NN	O	O
mutants	NN	O	O
of	NN	O	O
EBF	NN	O	B-protein
identified	NN	O	O
an	NN	O	O
amino-terminal	NN	O	B-protein
cysteine-rich	NN	O	I-protein
DNA-binding	NN	O	I-protein
domain	NN	O	I-protein
lacking	NN	O	O
obvious	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
to	NN	O	O
known	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

DNA-binding	NN	O	O
assays	NN	O	O
with	NN	O	O
cotranslated	NN	O	O
wild-type	NN	O	O
and	NN	O	O
truncated	NN	O	O
forms	NN	O	O
of	NN	O	O
EBF	NN	O	B-protein
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
protein	NN	O	O
interacts	NN	O	O
with	NN	O	O
its	NN	O	O
site	NN	O	O
as	NN	O	O
a	NN	O	O
homodimer	NN	O	B-protein
.	NN	O	O

Deletions	NN	O	O
delineated	NN	O	O
a	NN	O	O
carboxy-terminal	NN	O	B-protein
dimerization	NN	O	I-protein
region	NN	O	I-protein
containing	NN	O	O
two	NN	O	O
repeats	NN	O	O
of	NN	O	O
15	NN	O	O
amino	NN	O	O
acids	NN	O	O
that	NN	O	O
show	NN	O	O
similarity	NN	O	O
with	NN	O	O
the	NN	O	O
dimerization	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
basic-helix-loop-helix	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
EBF	NN	O	B-protein
represents	NN	O	O
a	NN	O	O
novel	NN	O	O
regulator	NN	O	O
of	NN	O	O
B	NN	O	O
lymphocyte-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cell-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
helix-loop-helix	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
E2A	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
E2A	NN	O	B-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
of	NN	O	O
the	NN	O	O
helix-loop-helix	NN	O	B-protein
family	NN	O	I-protein
that	NN	O	O
are	NN	O	O
implicated	NN	O	O
in	NN	O	O
cell-specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
as	NN	O	O
part	NN	O	O
of	NN	O	O
dimeric	NN	O	B-protein
complexes	NN	O	I-protein
that	NN	O	O
interact	NN	O	O
with	NN	O	O
E	NN	O	B-DNA
box	NN	O	I-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

It	NN	O	O
has	NN	O	O
previously	NN	O	O
been	NN	O	O
shown	NN	O	O
that	NN	O	O
transcripts	NN	O	O
of	NN	O	O
the	NN	O	O
E2A	NN	O	B-DNA
gene	NN	O	I-DNA
can	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
a	NN	O	O
wide	NN	O	O
range	NN	O	O
of	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
examined	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
E2A	NN	O	I-DNA
gene	NN	O	I-DNA
at	NN	O	O
the	NN	O	O
protein	NN	O	O
level	NN	O	O
using	NN	O	O
polyclonal	NN	O	O
antisera	NN	O	O
directed	NN	O	O
against	NN	O	O
distinct	NN	O	O
portions	NN	O	O
of	NN	O	O
the	NN	O	O
E2A	NN	O	B-protein
protein	NN	O	I-protein
to	NN	O	O
probe	NN	O	O
blots	NN	O	O
of	NN	O	O
cellular	NN	O	O
extracts	NN	O	O
.	NN	O	O

A	NN	O	O
73	NN	O	B-protein
kDa	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
identified	NN	O	O
by	NN	O	O
this	NN	O	O
analysis	NN	O	O
:	NN	O	O
this	NN	O	O
protein	NN	O	O
is	NN	O	O
highly	NN	O	O
enriched	NN	O	O
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
of	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
origin	NN	O	I-cell_line
as	NN	O	O
compared	NN	O	O
to	NN	O	O
pancreatic	NN	O	B-cell_type
beta-cells	NN	O	I-cell_type
and	NN	O	O
fibroblast	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
detection	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
selectively	NN	O	O
in	NN	O	O
extracts	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
E	NN	O	B-DNA
box	NN	O	I-DNA
-binding	NN	O	O
activity	NN	O	O
LEF1/BCF1	NN	O	B-protein
in	NN	O	O
these	NN	O	O
cells	NN	O	O
;	NN	O	O
this	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
previously	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
efficiently	NN	O	O
recognized	NN	O	O
by	NN	O	O
antiserum	NN	O	O
directed	NN	O	O
against	NN	O	O
E2A	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
full	NN	O	O
length	NN	O	O
E2A	NN	O	B-DNA
cDNA	NN	O	I-DNA
leads	NN	O	O
to	NN	O	O
appearance	NN	O	O
of	NN	O	O
protein	NN	O	O
co-migrating	NN	O	O
with	NN	O	O
the	NN	O	O
73	NN	O	B-protein
kDa	NN	O	I-protein
protein	NN	O	I-protein
on	NN	O	O
SDS	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
and	NN	O	O
co-migrating	NN	O	O
with	NN	O	O
LEF1/BCF1	NN	O	B-protein
on	NN	O	O
mobility	NN	O	O
shift	NN	O	O
analysis	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
view	NN	O	O
that	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
LEF1/BCF1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
homodimer	NN	O	B-protein
of	NN	O	I-protein
E2A	NN	O	I-protein
proteins	NN	O	I-protein
;	NN	O	O
the	NN	O	O
selective	NN	O	O
appearance	NN	O	O
of	NN	O	O
this	NN	O	O
putative	NN	O	O
cell-specific	NN	O	O
transcription	NN	O	O
factor	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
seems	NN	O	O
to	NN	O	O
be	NN	O	O
attributable	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
to	NN	O	O
the	NN	O	O
elevated	NN	O	O
E2A	NN	O	B-protein
protein	NN	O	I-protein
concentrations	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Hypertension	NN	O	O
in	NN	O	O
pregnancy	NN	O	O
.	NN	O	O

Pregnancy-induced	NN	O	O
hypertension	NN	O	O
(	NN	O	O
PIH	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
frequent	NN	O	O
cause	NN	O	O
of	NN	O	O
maternal	NN	O	O
and	NN	O	O
neonatal	NN	O	O
morbidity	NN	O	O
and	NN	O	O
mortality	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
focused	NN	O	O
on	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
PIH	NN	O	O
,	NN	O	O
mainly	NN	O	O
on	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
mineralocorticoids	NN	O	O
,	NN	O	O
reversed	NN	O	O
blood	NN	O	O
pressure	NN	O	O
patterns	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
resulting	NN	O	O
necessity	NN	O	O
of	NN	O	O
continuous	NN	O	O
monitoring	NN	O	O
of	NN	O	O
the	NN	O	O
preeclamptic	NN	O	O
mother	NN	O	O
.	NN	O	O

Problems	NN	O	O
of	NN	O	O
antihypertensive	NN	O	O
therapy	NN	O	O
are	NN	O	O
discussed	NN	O	O
and	NN	O	O
the	NN	O	O
first	NN	O	O
results	NN	O	O
of	NN	O	O
a	NN	O	O
pilot	NN	O	O
study	NN	O	O
with	NN	O	O
Urapidil	NN	O	O
are	NN	O	O
presented	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
mineralocorticoids	NN	O	O
in	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
PIH	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
plasma	NN	O	O
aldosterone	NN	O	O
and	NN	O	O
18-hydroxy-corticosterone	NN	O	O
(	NN	O	O
18-OH-B	NN	O	O
)	NN	O	O
levels	NN	O	O
in	NN	O	O
25	NN	O	O
women	NN	O	O
with	NN	O	O
PIH	NN	O	O
and	NN	O	O
in	NN	O	O
25	NN	O	O
healthy	NN	O	O
pregnant	NN	O	O
women	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
evaluated	NN	O	O
the	NN	O	O
mineralocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
MR	NN	O	B-protein
)	NN	O	O
count	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
2	NN	O	O
groups	NN	O	O
.	NN	O	O

The	NN	O	O
MR-count	NN	O	O
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
in	NN	O	O
the	NN	O	O
PIH-group	NN	O	O
.	NN	O	O

The	NN	O	O
values	NN	O	O
of	NN	O	O
plasma	NN	O	O
aldosterone	NN	O	O
and	NN	O	O
18-OH-B	NN	O	O
were	NN	O	O
also	NN	O	O
low	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
receptor	NN	O	O
down-regulation	NN	O	O
due	NN	O	O
to	NN	O	O
higher	NN	O	O
level	NN	O	O
of	NN	O	O
mineralocorticoids	NN	O	O
of	NN	O	O
the	NN	O	O
zona	NN	O	O
glomerulosa	NN	O	O
.	NN	O	O

Perhaps	NN	O	O
deoxycorticosterone	NN	O	O
or	NN	O	O
a	NN	O	O
hitherto	NN	O	O
unknown	NN	O	O
mineralocorticoid	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
hypertension	NN	O	O
and	NN	O	O
altered	NN	O	O
MR	NN	O	B-protein
-status	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
results	NN	O	O
of	NN	O	O
continuous	NN	O	O
blood	NN	O	O
pressure	NN	O	O
measurements	NN	O	O
with	NN	O	O
a	NN	O	O
noninvasive	NN	O	O
,	NN	O	O
real-time	NN	O	O
blood	NN	O	O
pressure	NN	O	O
monitor	NN	O	O
(	NN	O	O
Finapres	NN	O	O
)	NN	O	O
are	NN	O	O
presented	NN	O	O
.	NN	O	O

The	NN	O	O
comparison	NN	O	O
of	NN	O	O
the	NN	O	O
obtained	NN	O	O
values	NN	O	O
with	NN	O	O
intraarterial	NN	O	O
measurements	NN	O	O
demonstrates	NN	O	O
a	NN	O	O
good	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
methods	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
report	NN	O	O
on	NN	O	O
the	NN	O	O
first	NN	O	O
experiences	NN	O	O
with	NN	O	O
Urapidil	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
hypertension	NN	O	O
in	NN	O	O
severe	NN	O	O
preeclampsia	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
hypertension	NN	O	O
in	NN	O	O
preeclamptic	NN	O	O
women	NN	O	O
can	NN	O	O
be	NN	O	O
treated	NN	O	O
by	NN	O	O
Urapidil	NN	O	O
without	NN	O	O
side	NN	O	O
effects	NN	O	O
or	NN	O	O
reflex-tachycardia	NN	O	O
.	NN	O	O

Further	NN	O	O
studies	NN	O	O
will	NN	O	O
have	NN	O	O
to	NN	O	O
prove	NN	O	O
if	NN	O	O
Urapidil	NN	O	O
is	NN	O	O
suited	NN	O	O
for	NN	O	O
prepartal	NN	O	O
treatment	NN	O	O
of	NN	O	O
PIH	NN	O	O
as	NN	O	O
well	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
a	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
program	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
coincides	NN	O	O
with	NN	O	O
inhibition	NN	O	O
of	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
NF-E2	NN	O	B-protein
)	NN	O	O
to	NN	O	O
an	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
required	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
erythroid-specific	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Induction	NN	O	O
by	NN	O	O
hemin	NN	O	O
increases	NN	O	O
,	NN	O	O
while	NN	O	O
induction	NN	O	O
with	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
represses	NN	O	O
,	NN	O	O
erythroid-specific	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
K562	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
hemin	NN	O	O
or	NN	O	O
TPA	NN	O	O
induction	NN	O	O
on	NN	O	O
the	NN	O	O
binding	NN	O	O
and	NN	O	O
activity	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
at	NN	O	O
a	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
found	NN	O	O
within	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
regulatory	NN	O	I-DNA
sequences	NN	O	I-DNA
of	NN	O	O
many	NN	O	O
erythroid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

TPA	NN	O	O
induction	NN	O	O
increases	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
ubiquitous	NN	O	B-protein
AP-1	NN	O	I-protein
factors	NN	O	I-protein
to	NN	O	O
this	NN	O	O
element	NN	O	O
.	NN	O	O

TPA	NN	O	O
induction	NN	O	O
inhibits	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
lineage	NN	O	B-protein
limited	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-E2	NN	O	B-protein
to	NN	O	O
this	NN	O	O
transcriptional	NN	O	B-DNA
control	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Hemin	NN	O	O
induction	NN	O	O
of	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
does	NN	O	O
not	NN	O	O
facilitate	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-E2	NN	O	B-protein
to	NN	O	O
its	NN	O	O
recognition	NN	O	O
site	NN	O	O
.	NN	O	O

Hemin	NN	O	O
induction	NN	O	O
appears	NN	O	O
to	NN	O	O
nonspecifically	NN	O	O
increase	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
genes	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Beyond	NN	O	O
this	NN	O	O
nonspecific	NN	O	O
increase	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
hemin	NN	O	B-protein
induction	NN	O	O
acts	NN	O	O
to	NN	O	O
increase	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
lineage	NN	O	B-protein
limited	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-E2	NN	O	O
.	NN	O	O

The	NN	O	O
divergent	NN	O	O
effects	NN	O	O
of	NN	O	O
hemin	NN	O	O
and	NN	O	O
TPA	NN	O	O
on	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
mediated	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
their	NN	O	O
contrasting	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-E2	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Interleukin-3	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
involves	NN	O	O
an	NN	O	O
inducible	NN	O	O
,	NN	O	O
T-cell-specific	NN	O	B-protein
factor	NN	O	I-protein
and	NN	O	O
an	NN	O	O
octamer	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Interleukin-3	NN	O	B-protein
(	NN	O	O
IL-3	NN	O	B-protein
)	NN	O	O
is	NN	O	O
exclusively	NN	O	O
expressed	NN	O	O
by	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
and	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
function	NN	O	O
that	NN	O	O
is	NN	O	O
tightly	NN	O	O
controlled	NN	O	O
both	NN	O	O
in	NN	O	O
a	NN	O	O
lineage-specific	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
stimulation-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
protein	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
and	NN	O	O
functional	NN	O	O
importance	NN	O	O
of	NN	O	O
the	NN	O	O
ACT-1-activating	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-3	NN	O	B-protein
promoter	NN	O	O
.	NN	O	O

This	NN	O	O
region	NN	O	O
binds	NN	O	O
an	NN	O	O
inducible	NN	O	O
,	NN	O	O
T-cell-specific	NN	O	B-protein
factor	NN	O	I-protein
over	NN	O	O
its	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
end	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
site	NN	O	O
that	NN	O	O
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-3	NN	O	B-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
other	NN	O	O
upstream	NN	O	O
elements	NN	O	O
.	NN	O	O

Over	NN	O	O
its	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
end	NN	O	I-DNA
,	NN	O	O
it	NN	O	O
binds	NN	O	O
a	NN	O	O
factor	NN	O	O
that	NN	O	O
is	NN	O	O
ubiquitously	NN	O	O
and	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
.	NN	O	O

This	NN	O	O
factor	NN	O	O
is	NN	O	O
Oct-1	NN	O	B-protein
or	NN	O	O
an	NN	O	O
immunologically	NN	O	B-protein
related	NN	O	I-protein
octamer-binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
it	NN	O	O
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
coordinating	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
several	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
characteristics	NN	O	O
make	NN	O	O
the	NN	O	O
ACT-1	NN	O	B-DNA
site	NN	O	I-DNA
analogous	NN	O	O
to	NN	O	O
the	NN	O	O
activating	NN	O	O
ARRE-1	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
and	NN	O	O
despite	NN	O	O
a	NN	O	O
lack	NN	O	O
of	NN	O	O
sequence	NN	O	O
homology	NN	O	O
,	NN	O	O
the	NN	O	O
promoters	NN	O	O
of	NN	O	O
IL-3	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
share	NN	O	O
an	NN	O	O
organizational	NN	O	O
pattern	NN	O	O
of	NN	O	O
regulatory	NN	O	O
elements	NN	O	O
that	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
T-cell-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interleukin-4	NN	O	B-protein
inhibits	NN	O	O
the	NN	O	O
lipopolysaccharide-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
and	NN	O	I-RNA
c-fos	NN	O	I-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
and	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
studied	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
interleukin-4	NN	O	B-protein
(	NN	O	O
IL-4	NN	O	B-protein
)	NN	O	O
on	NN	O	O
the	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
induction	NN	O	O
of	NN	O	O
two	NN	O	O
immediate	NN	O	O
early	NN	O	O
genes	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	B-DNA
.	NN	O	O

These	NN	O	O
genes	NN	O	O
encode	NN	O	O
proteins	NN	O	O
that	NN	O	O
form	NN	O	O
the	NN	O	O
dimeric	NN	O	B-protein
complex	NN	O	I-protein
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
active	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

Maximal	NN	O	O
accumulation	NN	O	O
of	NN	O	O
either	NN	O	O
c-fos	NN	O	B-RNA
and	NN	O	I-RNA
c-jun	NN	O	I-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
(	NN	O	O
mRNA	NN	O	O
)	NN	O	O
occurred	NN	O	O
30	NN	O	O
minutes	NN	O	O
after	NN	O	O
LPS	NN	O	O
addition	NN	O	O
.	NN	O	O

When	NN	O	O
cells	NN	O	O
were	NN	O	O
treated	NN	O	O
with	NN	O	O
IL-4	NN	O	B-protein
for	NN	O	O
5	NN	O	O
hours	NN	O	O
before	NN	O	O
LPS	NN	O	O
activation	NN	O	O
,	NN	O	O
both	NN	O	O
the	NN	O	O
c-fos	NN	O	O
and	NN	O	O
the	NN	O	O
c-jun	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
was	NN	O	O
decreased	NN	O	O
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	O
expression	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
in	NN	O	O
LPS-treated	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
lower	NN	O	O
transcription	NN	O	O
rate	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	I-DNA
c-jun	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

IL-4	NN	O	B-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-protein
and	NN	O	O
c-jun	NN	O	B-protein
transcripts	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
evidence	NN	O	O
was	NN	O	O
obtained	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
inhibits	NN	O	O
LPS-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
protein	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
IL-4	NN	O	B-protein
suppresses	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
activated	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Transient	NN	O	O
pseudohypoaldosteronism	NN	O	O
in	NN	O	O
obstructive	NN	O	O
renal	NN	O	O
disease	NN	O	O
with	NN	O	O
transient	NN	O	O
reduction	NN	O	O
of	NN	O	O
lymphocytic	NN	O	B-protein
aldosterone	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Results	NN	O	O
in	NN	O	O
two	NN	O	O
affected	NN	O	O
infants	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
two	NN	O	O
patients	NN	O	O
with	NN	O	O
transient	NN	O	O
pseudohypoaldosteronism	NN	O	O
due	NN	O	O
to	NN	O	O
obstructive	NN	O	O
renal	NN	O	O
disease	NN	O	O
.	NN	O	O

Both	NN	O	O
patients	NN	O	O
presented	NN	O	O
with	NN	O	O
a	NN	O	O
salt-losing	NN	O	O
episode	NN	O	O
simulating	NN	O	O
adrenal	NN	O	O
insufficiency	NN	O	O
.	NN	O	O

In	NN	O	O
one	NN	O	O
patient	NN	O	O
,	NN	O	O
transient	NN	O	O
reduction	NN	O	O
of	NN	O	O
aldosterone	NN	O	B-protein
receptors	NN	O	I-protein
could	NN	O	O
be	NN	O	O
documented	NN	O	O
,	NN	O	O
while	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
patient	NN	O	O
the	NN	O	O
clinical	NN	O	O
and	NN	O	O
biochemical	NN	O	O
parameters	NN	O	O
were	NN	O	O
consistent	NN	O	O
with	NN	O	O
transient	NN	O	O
pseudohypoaldosteronism	NN	O	O
.	NN	O	O

Aldosterone	NN	O	B-protein
receptors	NN	O	I-protein
were	NN	O	O
normal	NN	O	O
in	NN	O	O
both	NN	O	O
patients	NN	O	O

-DOCSTART-	O

Aldosterone-specific	NN	O	B-protein
membrane	NN	O	I-protein
receptors	NN	O	I-protein
and	NN	O	O
rapid	NN	O	O
non-genomic	NN	O	O
actions	NN	O	O
of	NN	O	O
mineralocorticoids	NN	O	O
.	NN	O	O

Functional	NN	O	O
studies	NN	O	O
in	NN	O	O
extrarenal	NN	O	B-cell_type
,	NN	O	I-cell_type
non-epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
such	NN	O	O
as	NN	O	O
smooth	NN	O	B-cell_type
muscle	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
more	NN	O	O
recently	NN	O	O
circulating	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
have	NN	O	O
provided	NN	O	O
increasing	NN	O	O
evidence	NN	O	O
that	NN	O	O
aldosterone	NN	O	O
produces	NN	O	O
not	NN	O	O
only	NN	O	O
classical	NN	O	O
genomic	NN	O	O
effects	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
rapid	NN	O	O
,	NN	O	O
non-genomic	NN	O	O
effects	NN	O	O
on	NN	O	O
transmembrane	NN	O	O
electrolyte	NN	O	O
movements	NN	O	O
.	NN	O	O

These	NN	O	O
involve	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
sodium/proton	NN	O	B-protein
exchanger	NN	O	I-protein
of	NN	O	O
the	NN	O	O
cell	NN	O	O
membrane	NN	O	O
at	NN	O	O
very	NN	O	O
low	NN	O	O
,	NN	O	O
physiological	NN	O	O
concentrations	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
with	NN	O	O
an	NN	O	O
acute	NN	O	O
onset	NN	O	O
within	NN	O	O
1-2	NN	O	O
min	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
messenger	NN	O	O
cascade	NN	O	O
involved	NN	O	O
is	NN	O	O
the	NN	O	O
inositol	NN	O	O
1	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
5-trisphosphate/calcium	NN	O	O
pathway	NN	O	O
which	NN	O	O
responds	NN	O	O
over	NN	O	O
the	NN	O	O
same	NN	O	O
rapid	NN	O	O
time	NN	O	O
course	NN	O	O
.	NN	O	O

Such	NN	O	O
changes	NN	O	O
clearly	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
genomic	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
responsible	NN	O	O
for	NN	O	O
later	NN	O	O
effects	NN	O	O
than	NN	O	O
the	NN	O	O
membrane	NN	O	O
related	NN	O	O
rapid	NN	O	O
responses	NN	O	O
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
these	NN	O	O
rapid	NN	O	O
effects	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
on	NN	O	O
electrolytes	NN	O	O
have	NN	O	O
been	NN	O	O
extensively	NN	O	O
studied	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
thus	NN	O	O
may	NN	O	O
represent	NN	O	O
valuable	NN	O	O
tools	NN	O	O
in	NN	O	O
the	NN	O	O
delineation	NN	O	O
of	NN	O	O
the	NN	O	O
receptor-effector	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
.	NN	O	O

The	NN	O	O
unique	NN	O	O
characteristics	NN	O	O
of	NN	O	O
this	NN	O	O
new	NN	O	O
pathway	NN	O	O
for	NN	O	O
steroid	NN	O	O
action	NN	O	O
include	NN	O	O
its	NN	O	O
rapid	NN	O	O
time	NN	O	O
course	NN	O	O
,	NN	O	O
10	NN	O	O
,	NN	O	O
000-fold	NN	O	O
selectivity	NN	O	O
for	NN	O	O
aldosterone	NN	O	O
over	NN	O	O
cortisol	NN	O	O
and	NN	O	O
the	NN	O	O
ineffectiveness	NN	O	O
of	NN	O	O
spironolactones	NN	O	O
,	NN	O	O
classical	NN	O	O
mineralocorticoid	NN	O	O
antagonists	NN	O	O
,	NN	O	O
as	NN	O	O
antagonists	NN	O	O
of	NN	O	O
the	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-protein
immunodeficiency	NN	O	I-protein
virus	NN	O	I-protein
type	NN	O	I-protein
1	NN	O	I-protein
Nef	NN	O	I-protein
protein	NN	O	I-protein
inhibits	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
induction	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
can	NN	O	O
establish	NN	O	O
a	NN	O	O
persistent	NN	O	O
and	NN	O	O
latent	NN	O	O
infection	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
W.C.Greene	NN	O	O
,	NN	O	O
N.Engl.J.	NN	O	O
Med.324	NN	O	O
:	NN	O	O
308-317	NN	O	O
,	NN	O	O
1991	NN	O	O
;	NN	O	O
S.M.Schnittman	NN	O	O
,	NN	O	O
M.C.Psallidopoulos	NN	O	O
,	NN	O	O
H.C.	NN	O	O
Lane	NN	O	O
,	NN	O	O
L.Thompson	NN	O	O
,	NN	O	O
M.Baseler	NN	O	O
,	NN	O	O
F.Massari	NN	O	O
,	NN	O	O
C.H.Fox	NN	O	O
,	NN	O	O
N.P.Salzman	NN	O	O
,	NN	O	O
and	NN	O	O
A.S.Fauci	NN	O	O
,	NN	O	O
Science	NN	O	O
245	NN	O	O
:	NN	O	O
305-308	NN	O	O
,	NN	O	O
1989	NN	O	O
)	NN	O	O
.	NN	O	O

Production	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
from	NN	O	O
latently	NN	O	O
infected	NN	O	O
cells	NN	O	O
requires	NN	O	O
host	NN	O	O
cell	NN	O	O
activation	NN	O	O
by	NN	O	O
T-cell	NN	O	B-protein
mitogens	NN	O	I-protein
(	NN	O	O
T.Folks	NN	O	O
,	NN	O	O
D.M.Powell	NN	O	O
,	NN	O	O
M.M.Lightfoote	NN	O	O
,	NN	O	O
S.Benn	NN	O	O
,	NN	O	O
M.A.	NN	O	O
Martin	NN	O	O
,	NN	O	O
and	NN	O	O
A.S.Fauci	NN	O	O
,	NN	O	O
Science	NN	O	O
231	NN	O	O
:	NN	O	O
600-602	NN	O	O
,	NN	O	O
1986	NN	O	O
;	NN	O	O
D.Zagury	NN	O	O
,	NN	O	O
J.	NN	O	O
Bernard	NN	O	O
,	NN	O	O
R.Leonard	NN	O	O
,	NN	O	O
R.Cheynier	NN	O	O
,	NN	O	O
M.Feldman	NN	O	O
,	NN	O	O
P.S.Sarin	NN	O	O
,	NN	O	O
and	NN	O	O
R.C.	NN	O	O
Gallo	NN	O	O
,	NN	O	O
Science	NN	O	O
231	NN	O	O
:	NN	O	O
850-853	NN	O	O
,	NN	O	O
1986	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
activation	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
host	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
[	NN	O	O
G.Nabel	NN	O	O
and	NN	O	O
D.Baltimore	NN	O	O
,	NN	O	O
Nature	NN	O	O
(	NN	O	O
London	NN	O	O
)	NN	O	O
326	NN	O	O
:	NN	O	O
711-717	NN	O	O
,	NN	O	O
1987	NN	O	O
]	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
HIV-1-encoded	NN	O	B-protein
Nef	NN	O	I-protein
protein	NN	O	I-protein
inhibits	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA-binding	NN	O	O
activity	NN	O	O
by	NN	O	O
T-	NN	O	B-protein
cell	NN	O	I-protein
mitogens	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
Nef	NN	O	B-protein
does	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
implicated	NN	O	O
in	NN	O	O
HIV-1	NN	O	O
regulation	NN	O	O
,	NN	O	O
including	NN	O	O
SP-1	NN	O	B-protein
,	NN	O	O
USF	NN	O	B-protein
,	NN	O	O
URS	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
Nef	NN	O	B-protein
inhibits	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV-1-	NN	O	O
and	NN	O	O
interleukin	NN	O	O
2-directed	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
effect	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
depends	NN	O	O
on	NN	O	O
an	NN	O	O
intact	NN	O	O
NF-kappa	NN	O	B-DNA
B-binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
defective	NN	O	O
recruitment	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
may	NN	O	O
underlie	NN	O	O
Nef	NN	O	B-protein
's	NN	O	O
negative	NN	O	O
transcriptional	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
and	NN	O	I-DNA
interleukin	NN	O	I-DNA
2	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

Further	NN	O	O
evidence	NN	O	O
suggests	NN	O	O
that	NN	O	O
Nef	NN	O	B-protein
inhibits	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
induction	NN	O	O
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
a	NN	O	O
signal	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
,	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
jun	NN	O	B-DNA
B	NN	O	I-DNA
in	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
lymphocytes	NN	O	O
from	NN	O	O
young	NN	O	O
and	NN	O	O
elderly	NN	O	O
adults	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
,	NN	O	I-DNA
c-jun	NN	O	I-DNA
and	NN	O	I-DNA
jun	NN	O	I-DNA
B	NN	O	I-DNA
proto-oncogenes	NN	O	I-DNA
was	NN	O	O
studied	NN	O	O
in	NN	O	O
phytohemagglutinin	NN	O	B-cell_line
(	NN	O	I-cell_line
PHA	NN	O	I-cell_line
)	NN	O	I-cell_line
activated	NN	O	I-cell_line
peripheral	NN	O	I-cell_line
blood	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
(	NN	O	O
PBL	NN	O	B-cell_type
)	NN	O	O
from	NN	O	O
young	NN	O	O
and	NN	O	O
aged	NN	O	O
humans	NN	O	O
.	NN	O	O

Specific	NN	O	O
mRNAs	NN	O	B-RNA
for	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	B-DNA
were	NN	O	O
detectable	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
after	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
reached	NN	O	O
maximal	NN	O	O
levels	NN	O	O
within	NN	O	O
2	NN	O	O
h	NN	O	O
.	NN	O	O

Both	NN	O	O
c-fos	NN	O	B-RNA
and	NN	O	I-RNA
jun	NN	O	I-RNA
B	NN	O	I-RNA
mRNAs	NN	O	I-RNA
decreased	NN	O	O
to	NN	O	O
pre-activation	NN	O	O
levels	NN	O	O
within	NN	O	O
6	NN	O	O
h	NN	O	O
,	NN	O	O
while	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
remained	NN	O	O
elevated	NN	O	O
.	NN	O	O

In	NN	O	O
PHA-activated	NN	O	B-cell_line
PBL	NN	O	I-cell_line
,	NN	O	O
no	NN	O	O
age-related	NN	O	O
differences	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
c-fos	NN	O	O
or	NN	O	O
jun	NN	O	O
B	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
decreased	NN	O	O
significantly	NN	O	O
(	NN	O	O
1.73	NN	O	O
+/-	NN	O	O
0.08	NN	O	O
vs.	NN	O	O
1.16	NN	O	O
+/-	NN	O	O
0.09	NN	O	O
arbitrary	NN	O	O
units	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.01	NN	O	O
,	NN	O	O
young	NN	O	O
vs.	NN	O	O
old	NN	O	O
)	NN	O	O
in	NN	O	O
PBL	NN	O	B-cell_type
from	NN	O	O
elderly	NN	O	O
individuals	NN	O	O
activated	NN	O	O
with	NN	O	O
PHA	NN	O	B-protein
.	NN	O	O

Because	NN	O	O
previous	NN	O	O
work	NN	O	O
has	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
elderly	NN	O	O
individuals	NN	O	O
may	NN	O	O
display	NN	O	O
normal	NN	O	O
proliferative	NN	O	O
responses	NN	O	O
when	NN	O	O
activated	NN	O	O
via	NN	O	O
the	NN	O	O
anti-CD2	NN	O	B-protein
pathway	NN	O	O
,	NN	O	O
c-jun	NN	O	O
and	NN	O	O
jun	NN	O	O
B	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
was	NN	O	O
also	NN	O	O
studied	NN	O	O
in	NN	O	O
anti-CD2-activated	NN	O	B-cell_line
purified	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

No	NN	O	O
age-related	NN	O	O
differences	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
either	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
proto-oncogenes	NN	O	B-DNA
by	NN	O	O
anti-CD2	NN	O	O
activated	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
decreased	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
and	NN	O	O
proliferative	NN	O	O
response	NN	O	O
displayed	NN	O	O
by	NN	O	O
PHA-activated	NN	O	B-cell_line
PBL	NN	O	I-cell_line
from	NN	O	O
elderly	NN	O	O
adults	NN	O	O
may	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
age-related	NN	O	O
changes	NN	O	O
in	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
ratio	NN	O	O
of	NN	O	O
c-fos	NN	O	O
to	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

-DOCSTART-	O

Oct2	NN	O	B-protein
transactivation	NN	O	O
from	NN	O	O
a	NN	O	O
remote	NN	O	B-DNA
enhancer	NN	O	I-DNA
position	NN	O	I-DNA
requires	NN	O	O
a	NN	O	O
B-cell-restricted	NN	O	O
activity	NN	O	O
.	NN	O	O

Previous	NN	O	O
cotransfection	NN	O	O
experiments	NN	O	O
had	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
lymphocyte-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Oct2	NN	O	B-protein
could	NN	O	O
efficiently	NN	O	O
activate	NN	O	O
a	NN	O	O
promoter	NN	O	B-DNA
containing	NN	O	O
an	NN	O	O
octamer	NN	O	O
motif	NN	O	O
.	NN	O	O

Oct2	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
unable	NN	O	O
to	NN	O	O
stimulate	NN	O	O
a	NN	O	O
multimerized	NN	O	B-DNA
octamer	NN	O	I-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
however	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
tested	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
Oct2	NN	O	B-protein
isoforms	NN	O	I-protein
generated	NN	O	O
by	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
for	NN	O	O
the	NN	O	O
capability	NN	O	O
to	NN	O	O
activate	NN	O	O
an	NN	O	O
octamer	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
nonlymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
a	NN	O	O
B-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
analyses	NN	O	O
show	NN	O	O
that	NN	O	O
several	NN	O	O
Oct2	NN	O	B-protein
isoforms	NN	O	I-protein
can	NN	O	O
stimulate	NN	O	O
from	NN	O	O
a	NN	O	O
remote	NN	O	O
position	NN	O	O
but	NN	O	O
that	NN	O	O
this	NN	O	O
stimulation	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
result	NN	O	O
indicates	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
either	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	B-protein
cofactor	NN	O	I-protein
or	NN	O	O
a	NN	O	O
specific	NN	O	O
modification	NN	O	O
of	NN	O	O
a	NN	O	O
cofactor	NN	O	B-protein
or	NN	O	O
the	NN	O	O
Oct2	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
Oct2	NN	O	B-protein
-mediated	NN	O	O
enhancer	NN	O	O
activation	NN	O	O
.	NN	O	O

Mutational	NN	O	O
analyses	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
carboxy-terminal	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
Oct2	NN	O	B-protein
is	NN	O	O
critical	NN	O	O
for	NN	O	O
enhancer	NN	O	O
activation	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
this	NN	O	O
domain	NN	O	O
conferred	NN	O	O
enhancing	NN	O	O
activity	NN	O	O
when	NN	O	O
fused	NN	O	O
to	NN	O	O
the	NN	O	O
Oct1	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
by	NN	O	O
itself	NN	O	O
was	NN	O	O
unable	NN	O	O
to	NN	O	O
stimulate	NN	O	O
from	NN	O	O
a	NN	O	O
remote	NN	O	O
position	NN	O	O
.	NN	O	O

The	NN	O	O
glutamine-rich	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
present	NN	O	O
in	NN	O	O
the	NN	O	O
amino-terminal	NN	O	B-protein
portion	NN	O	I-protein
of	NN	O	I-protein
Oct2	NN	O	I-protein
and	NN	O	O
the	NN	O	O
POU	NN	O	B-protein
domain	NN	O	I-protein
contribute	NN	O	O
only	NN	O	O
marginally	NN	O	O
to	NN	O	O
the	NN	O	O
transactivation	NN	O	O
function	NN	O	O
from	NN	O	O
a	NN	O	O
distal	NN	O	O
position	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
CD19	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
BSAP	NN	O	B-protein
.	NN	O	O

The	NN	O	O
CD19	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
all	NN	O	O
B-lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
terminally	NN	O	B-cell_type
differentiated	NN	O	I-cell_type
plasma	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
as	NN	O	O
a	NN	O	O
signal-transducing	NN	O	O
receptor	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
proliferation	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
complete	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
expression	NN	O	O
pattern	NN	O	O
of	NN	O	O
the	NN	O	O
CD19	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
BSAP	NN	O	B-protein
in	NN	O	O
a	NN	O	O
large	NN	O	O
panel	NN	O	O
of	NN	O	O
B-lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
CD19	NN	O	I-DNA
gene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
cloned	NN	O	O
,	NN	O	O
and	NN	O	O
several	NN	O	O
BSAP-binding	NN	O	B-DNA
sites	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
mapped	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
protein-DNA	NN	O	O
binding	NN	O	O
studies	NN	O	O
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
a	NN	O	O
high-affinity	NN	O	B-DNA
BSAP-binding	NN	O	I-DNA
site	NN	O	I-DNA
instead	NN	O	O
of	NN	O	O
a	NN	O	O
TATA	NN	O	B-DNA
sequence	NN	O	I-DNA
is	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
-30	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
a	NN	O	O
cluster	NN	O	O
of	NN	O	O
heterogeneous	NN	O	O
transcription	NN	O	O
start	NN	O	O
sites	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
this	NN	O	O
site	NN	O	O
is	NN	O	O
occupied	NN	O	O
by	NN	O	O
BSAP	NN	O	B-protein
in	NN	O	O
vivo	NN	O	O
in	NN	O	O
a	NN	O	O
CD19-expressing	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
plasma	NN	O	B-cell_type
or	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
high-affinity	NN	O	B-DNA
site	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
conserved	NN	O	O
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
both	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
mouse	NN	O	I-DNA
CD19	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
was	NN	O	O
furthermore	NN	O	O
shown	NN	O	O
to	NN	O	O
confer	NN	O	O
B-cell	NN	O	O
specificity	NN	O	O
to	NN	O	O
a	NN	O	O
beta-globin	NN	O	O
reporter	NN	O	O
gene	NN	O	O
in	NN	O	O
transient	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
BSAP	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
only	NN	O	O
abundant	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
B-cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
that	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
CD19	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Together	NN	O	O
,	NN	O	O
this	NN	O	O
evidence	NN	O	O
strongly	NN	O	O
implicates	NN	O	O
BSAP	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
CD19	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Eicosanoids	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
mastectomy	NN	O	O
.	NN	O	O

In	NN	O	O
19	NN	O	O
patients	NN	O	O
with	NN	O	O
a	NN	O	O
malignant	NN	O	O
breast	NN	O	O
tumor	NN	O	O
,	NN	O	O
tumor	NN	O	O
tissue	NN	O	O
and	NN	O	O
blood	NN	O	O
were	NN	O	O
taken	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
eicosanoid	NN	O	O
profile	NN	O	O
and	NN	O	O
platelet	NN	O	B-cell_type
aggregation	NN	O	O
.	NN	O	O

Values	NN	O	O
were	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
benign	NN	O	O
tumors	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
4	NN	O	O
)	NN	O	O
,	NN	O	O
or	NN	O	O
undergoing	NN	O	O
a	NN	O	O
mammary	NN	O	O
reduction	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
7	NN	O	O
)	NN	O	O
.	NN	O	O

Postoperatively	NN	O	O
,	NN	O	O
blood	NN	O	O
was	NN	O	O
taken	NN	O	O
as	NN	O	O
well	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
compare	NN	O	O
pre-	NN	O	O
and	NN	O	O
postoperative	NN	O	O
values	NN	O	O
.	NN	O	O

Eicosanoids	NN	O	O
were	NN	O	O
measured	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
mammary	NN	O	O
tissue	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
HPLC	NN	O	O
;	NN	O	O
furthermore	NN	O	O
,	NN	O	O
TXA2	NN	O	O
,	NN	O	O
6-keto-PGF1	NN	O	O
alpha	NN	O	O
,	NN	O	O
and	NN	O	O
PGE2	NN	O	O
were	NN	O	O
determined	NN	O	O
by	NN	O	O
RIA	NN	O	O
.	NN	O	O

Differences	NN	O	O
in	NN	O	O
pre-	NN	O	O
and	NN	O	O
postoperative	NN	O	O
values	NN	O	O
of	NN	O	O
cancer	NN	O	O
patients	NN	O	O
were	NN	O	O
seen	NN	O	O
in	NN	O	O
plasma	NN	O	O
RIA	NN	O	O
values	NN	O	O
:	NN	O	O
PGE2	NN	O	O
and	NN	O	O
6-k-PGF1	NN	O	O
alpha	NN	O	O
were	NN	O	O
significantly	NN	O	O
higher	NN	O	O
preoperatively	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
postoperatively	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
such	NN	O	O
differences	NN	O	O
were	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
groups	NN	O	O
as	NN	O	O
well	NN	O	O
.	NN	O	O

Compared	NN	O	O
to	NN	O	O
benign	NN	O	O
tumor	NN	O	O
or	NN	O	O
mammary	NN	O	O
reduction	NN	O	O
test	NN	O	O
material	NN	O	O
the	NN	O	O
eicosanoid	NN	O	O
profile	NN	O	O
of	NN	O	O
tissue	NN	O	O
obtained	NN	O	O
from	NN	O	O
malignant	NN	O	O
mammary	NN	O	O
tumors	NN	O	O
showed	NN	O	O
important	NN	O	O
differences	NN	O	O
.	NN	O	O

Except	NN	O	O
for	NN	O	O
PGF2	NN	O	O
alpha	NN	O	O
,	NN	O	O
HHT	NN	O	O
and	NN	O	O
15-HETE	NN	O	O
no	NN	O	O
detectable	NN	O	O
quantities	NN	O	O
of	NN	O	O
eicosanoids	NN	O	O
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
non-tumor	NN	O	O
material	NN	O	O
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
the	NN	O	O
malignant	NN	O	O
tumor	NN	O	O
material	NN	O	O
substantial	NN	O	O
quantities	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
eicosanoid	NN	O	O
metabolites	NN	O	O
were	NN	O	O
present	NN	O	O
.	NN	O	O

Statistically	NN	O	O
significant	NN	O	O
correlations	NN	O	O
could	NN	O	O
be	NN	O	O
established	NN	O	O
between	NN	O	O
patient/histopathology	NN	O	O
data	NN	O	O
and	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
the	NN	O	O
platelet	NN	O	B-cell_type
aggregation	NN	O	O
assays	NN	O	O
,	NN	O	O
e.g.	NN	O	O
between	NN	O	O
menopausal	NN	O	O
status	NN	O	O
and	NN	O	O
ADP	NN	O	O
aggregation	NN	O	O
;	NN	O	O
oestrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
+/-	NN	O	O
)	NN	O	O
and	NN	O	O
collagen	NN	O	B-protein
and	NN	O	O
arachidonic	NN	O	O
acid	NN	O	O
aggregation	NN	O	O
,	NN	O	O
inflammatory	NN	O	O
cell	NN	O	O
infiltration	NN	O	O
score	NN	O	O
and	NN	O	O
arachidonic	NN	O	O
acid	NN	O	O
aggregation	NN	O	O
and	NN	O	O
fibrosis	NN	O	O
score	NN	O	O
and	NN	O	O
ADP	NN	O	O
aggregation	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
eicosanoid	NN	O	O
synthesis	NN	O	O
in	NN	O	O
material	NN	O	O
from	NN	O	O
mammary	NN	O	O
cancer	NN	O	O
patients	NN	O	O
is	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
in	NN	O	O
benign	NN	O	O
mammary	NN	O	O
tissue	NN	O	O
.	NN	O	O

The	NN	O	O
implications	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
future	NN	O	O
prognosis	NN	O	O
of	NN	O	O
the	NN	O	O
patient	NN	O	O
,	NN	O	O
remain	NN	O	O
obscure	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mineralocorticoids	NN	O	O
and	NN	O	O
mineralocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
pregnancy-induced	NN	O	O
hypertension	NN	O	O
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
mineralocorticoids	NN	O	O
in	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
pregnancy-induced	NN	O	O
hypertension	NN	O	O
(	NN	O	O
PIH	NN	O	O
)	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
plasma	NN	O	O
aldosterone	NN	O	O
and	NN	O	O
18-hydroxycorticosterone	NN	O	O
levels	NN	O	O
in	NN	O	O
25	NN	O	O
women	NN	O	O
with	NN	O	O
PIH	NN	O	O
and	NN	O	O
25	NN	O	O
normal	NN	O	O
pregnant	NN	O	O
women	NN	O	O
,	NN	O	O
as	NN	O	O
controls	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
evaluated	NN	O	O
the	NN	O	O
mineralocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
MR	NN	O	B-protein
)	NN	O	O
status	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
2	NN	O	O
groups	NN	O	O
.	NN	O	O

MR	NN	O	B-protein
count	NN	O	O
was	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.0005	NN	O	O
)	NN	O	O
decreased	NN	O	O
in	NN	O	O
the	NN	O	O
PIH	NN	O	O
group	NN	O	O
(	NN	O	O
148	NN	O	O
+/-	NN	O	O
9	NN	O	O
binding	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
control	NN	O	O
group	NN	O	O
(	NN	O	O
300	NN	O	O
+/-	NN	O	O
17	NN	O	O
binding	NN	O	O
sites/cell	NN	O	O
;	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
)	NN	O	O
.	NN	O	O

Plasma	NN	O	O
aldosterone	NN	O	O
in	NN	O	O
women	NN	O	O
with	NN	O	O
PIH	NN	O	O
was	NN	O	O
281	NN	O	O
+/-	NN	O	O
61	NN	O	O
pmol/L	NN	O	O
;	NN	O	O
in	NN	O	O
normal	NN	O	O
pregnant	NN	O	O
women	NN	O	O
it	NN	O	O
was	NN	O	O
697	NN	O	O
+/-	NN	O	O
172	NN	O	O
pmol/L	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.025	NN	O	O
)	NN	O	O
.	NN	O	O

Plasma	NN	O	O
18-hydroxycorticosterone	NN	O	O
was	NN	O	O
also	NN	O	O
significantly	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.025	NN	O	O
)	NN	O	O
lower	NN	O	O
(	NN	O	O
PIH	NN	O	O
,	NN	O	O
1071	NN	O	O
+/-	NN	O	O
149	NN	O	O
pmol/L	NN	O	O
;	NN	O	O
controls	NN	O	O
,	NN	O	O
1907	NN	O	O
+/-	NN	O	O
318	NN	O	O
pmol/L	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
values	NN	O	O
were	NN	O	O
determined	NN	O	O
at	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
clinical	NN	O	O
symptoms	NN	O	O
of	NN	O	O
PIH	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
receptor	NN	O	O
down-regulation	NN	O	O
due	NN	O	O
to	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
mineralocorticoids	NN	O	O
in	NN	O	O
PIH	NN	O	O
;	NN	O	O
a	NN	O	O
hitherto	NN	O	O
unknown	NN	O	O
mineralocorticoid	NN	O	O
may	NN	O	O
,	NN	O	O
thus	NN	O	O
,	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
hypertension	NN	O	O
and	NN	O	O
altered	NN	O	O
MR	NN	O	B-protein
status	NN	O	O
.	NN	O	O

-DOCSTART-	O

Structure	NN	O	O
function	NN	O	O
analysis	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D	NN	O	O
analogs	NN	O	O
with	NN	O	O
C-ring	NN	O	O
modifications	NN	O	O
.	NN	O	O

Analogs	NN	O	O
of	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
)	NN	O	O
with	NN	O	O
substitutions	NN	O	O
on	NN	O	O
C-11	NN	O	O
were	NN	O	O
synthesized	NN	O	O
.	NN	O	O

Small	NN	O	O
apolar	NN	O	O
substitutions	NN	O	O
(	NN	O	O
11	NN	O	O
alpha-methyl	NN	O	O
,	NN	O	O
11	NN	O	O
alpha-fluoromethyl	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
markedly	NN	O	O
decrease	NN	O	O
the	NN	O	O
affinity	NN	O	O
for	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
but	NN	O	O
larger	NN	O	O
(	NN	O	O
11	NN	O	O
alpha-chloromethyl	NN	O	O
or	NN	O	O
11	NN	O	O
alpha-	NN	O	O
or	NN	O	O
11	NN	O	O
beta-phenyl	NN	O	O
)	NN	O	O
or	NN	O	O
more	NN	O	O
polar	NN	O	O
substitutions	NN	O	O
(	NN	O	O
11	NN	O	O
alpha-hydroxymethyl	NN	O	O
,	NN	O	O
11	NN	O	O
alpha-	NN	O	O
(	NN	O	O
2-hydroxyethyl	NN	O	O
]	NN	O	O
decreased	NN	O	O
the	NN	O	O
affinity	NN	O	O
to	NN	O	O
less	NN	O	O
than	NN	O	O
5	NN	O	O
%	NN	O	O
of	NN	O	O
that	NN	O	O
of	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
.	NN	O	O

Their	NN	O	O
affinity	NN	O	O
for	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D-binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
however	NN	O	O
,	NN	O	O
increased	NN	O	O
up	NN	O	O
to	NN	O	O
4-fold	NN	O	O
.	NN	O	O

The	NN	O	O
biological	NN	O	O
activity	NN	O	O
of	NN	O	O
11	NN	O	O
alpha-methyl-1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
closely	NN	O	O
resembled	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
natural	NN	O	O
hormone	NN	O	O
on	NN	O	O
normal	NN	O	O
and	NN	O	O
leukemic	NN	O	B-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
and	NN	O	O
bone	NN	O	O
resorption	NN	O	O
,	NN	O	O
whereas	NN	O	O
its	NN	O	O
in	NN	O	O
vivo	NN	O	O
effect	NN	O	O
on	NN	O	O
calcium	NN	O	O
metabolism	NN	O	O
of	NN	O	O
the	NN	O	O
rachitic	NN	O	O
chick	NN	O	O
was	NN	O	O
about	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
that	NN	O	O
of	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
.	NN	O	O

The	NN	O	O
11	NN	O	O
beta-methyl	NN	O	O
analog	NN	O	O
had	NN	O	O
a	NN	O	O
greater	NN	O	O
than	NN	O	O
10-fold	NN	O	O
lower	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
differentiating	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
C-11	NN	O	O
analogs	NN	O	O
on	NN	O	O
human	NN	O	B-cell_line
promyeloid	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
HL-60	NN	O	B-cell_line
)	NN	O	O
agreed	NN	O	O
well	NN	O	O
with	NN	O	O
their	NN	O	O
bone-resorbing	NN	O	O
activity	NN	O	O
and	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
,	NN	O	O
but	NN	O	O
they	NN	O	O
demonstrated	NN	O	O
lower	NN	O	O
calcemic	NN	O	O
effects	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Large	NN	O	O
or	NN	O	O
polar	NN	O	O
substitutions	NN	O	O
on	NN	O	O
C-11	NN	O	O
of	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
thus	NN	O	O
impair	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
but	NN	O	O
increase	NN	O	O
the	NN	O	O
affinity	NN	O	O
to	NN	O	O
vitamin	NN	O	B-protein
D-binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
many	NN	O	O
C-11-substituted	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
analogs	NN	O	O
on	NN	O	O
HL-60	NN	O	B-cell_line
cell	NN	O	O
differentiation	NN	O	O
exceeded	NN	O	O
their	NN	O	O
activity	NN	O	O
on	NN	O	O
calcium	NN	O	O
metabolism	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
in	NN	O	O
three	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
in	NN	O	O
major	NN	O	O
depressive	NN	O	O
disorder	NN	O	O
:	NN	O	O
lack	NN	O	O
of	NN	O	O
evidence	NN	O	O
of	NN	O	O
receptor	NN	O	O
binding	NN	O	O
defect	NN	O	O
.	NN	O	O

1	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
further	NN	O	O
understand	NN	O	O
the	NN	O	O
apparent	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
in	NN	O	O
major	NN	O	O
depressive	NN	O	O
disorder	NN	O	O
,	NN	O	O
circadian	NN	O	O
variation	NN	O	O
in	NN	O	O
cortisol	NN	O	O
concentration	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
suppression	NN	O	O
and	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
binding	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
and	NN	O	O
cultured	NN	O	B-cell_line
skin	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
were	NN	O	O
measured	NN	O	O
in	NN	O	O
rigidly	NN	O	O
defined	NN	O	O
major	NN	O	O
depressive	NN	O	O
disorder	NN	O	O
patients	NN	O	O
and	NN	O	O
non-depressed	NN	O	O
psychiatric	NN	O	O
controls	NN	O	O
.	NN	O	O

2	NN	O	O
.	NN	O	O

Mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
binding	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
correlated	NN	O	O
significantly	NN	O	O
with	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
binding	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
,	NN	O	O
but	NN	O	O
both	NN	O	O
determinations	NN	O	O
failed	NN	O	O
to	NN	O	O
differentiate	NN	O	O
major	NN	O	O
depressive	NN	O	O
disorder	NN	O	O
and	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

3	NN	O	O
.	NN	O	O

Initial	NN	O	O
and	NN	O	O
post-dexamethasone	NN	O	O
in	NN	O	O
vitro	NN	O	O
fibroblast	NN	O	O
binding	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
was	NN	O	O
not	NN	O	O
different	NN	O	O
between	NN	O	O
major	NN	O	O
depressive	NN	O	O
disorder	NN	O	O
and	NN	O	O
non-depressed	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

4	NN	O	O
.	NN	O	O

The	NN	O	O
phenomenon	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
in	NN	O	O
major	NN	O	O
depressive	NN	O	O
disorder	NN	O	O
remains	NN	O	O
unexplained	NN	O	O
.	NN	O	O

-DOCSTART-	O

High	NN	O	O
affinity	NN	O	O
aldosterone	NN	O	O
binding	NN	O	O
to	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
rich	NN	O	O
fractions	NN	O	O
from	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
:	NN	O	O
is	NN	O	O
there	NN	O	O
a	NN	O	O
membrane	NN	O	B-protein
receptor	NN	O	I-protein
for	NN	O	O
mineralocorticoids	NN	O	O
?	NN	O	O

In	NN	O	O
vitro	NN	O	O
effects	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
on	NN	O	O
the	NN	O	O
intracellular	NN	O	O
concentrations	NN	O	O
of	NN	O	O
sodium	NN	O	O
,	NN	O	O
potassium	NN	O	O
and	NN	O	O
calcium	NN	O	O
,	NN	O	O
cell	NN	O	O
volume	NN	O	O
and	NN	O	O
the	NN	O	O
sodium-proton-antiport	NN	O	B-protein
have	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
intact	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
HML	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
paper	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
[	NN	O	O
125I	NN	O	O
]	NN	O	O
-labeled	NN	O	O
aldosterone	NN	O	O
derivative	NN	O	O
to	NN	O	O
plasma	NN	O	O
membrane	NN	O	O
rich	NN	O	O
fractions	NN	O	O
of	NN	O	O
HML	NN	O	B-cell_type
was	NN	O	O
studied	NN	O	O
.	NN	O	O

High	NN	O	O
affinity	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
radioligand	NN	O	O
with	NN	O	O
an	NN	O	O
apparent	NN	O	O
Kd	NN	O	O
of	NN	O	O
approximately	NN	O	O
0.1	NN	O	O
nM	NN	O	O
was	NN	O	O
found	NN	O	O
.	NN	O	O

Aldosterone	NN	O	O
displaced	NN	O	O
the	NN	O	O
tracer	NN	O	O
at	NN	O	O
a	NN	O	O
similar	NN	O	O
Kd	NN	O	O
.	NN	O	O

Both	NN	O	O
canrenone	NN	O	O
and	NN	O	O
cortisol	NN	O	O
were	NN	O	O
inactive	NN	O	O
as	NN	O	O
ligands	NN	O	O
up	NN	O	O
to	NN	O	O
concentrations	NN	O	O
of	NN	O	O
0.1	NN	O	O
microM	NN	O	O
.	NN	O	O

The	NN	O	O
findings	NN	O	O
are	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
membrane	NN	O	O
binding	NN	O	O
sites	NN	O	O
with	NN	O	O
a	NN	O	O
high	NN	O	O
affinity	NN	O	O
for	NN	O	O
aldosterone	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
for	NN	O	O
cortisol	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
perfectly	NN	O	O
compatible	NN	O	O
with	NN	O	O
major	NN	O	O
properties	NN	O	O
of	NN	O	O
steroidal	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
sodium-proton-antiport	NN	O	B-protein
in	NN	O	O
HML	NN	O	B-cell_type
and	NN	O	O
thus	NN	O	O
very	NN	O	O
likely	NN	O	O
represent	NN	O	O
membrane	NN	O	B-protein
receptors	NN	O	I-protein
for	NN	O	O
aldosterone	NN	O	O
.	NN	O	O

-DOCSTART-	O

Natural	NN	O	O
variants	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
:	NN	O	O
analysis	NN	O	O
of	NN	O	O
promoters	NN	O	B-DNA
with	NN	O	O
duplicated	NN	O	O
DNA	NN	O	B-DNA
regulatory	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

Sequence	NN	O	O
variation	NN	O	O
in	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	I-DNA
LTR	NN	O	I-DNA
)	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
HIV-1	NN	O	O
was	NN	O	O
analyzed	NN	O	O
in	NN	O	O
viral	NN	O	O
isolates	NN	O	O
of	NN	O	O
17	NN	O	O
infected	NN	O	O
individuals	NN	O	O
.	NN	O	O

Two	NN	O	O
classes	NN	O	O
of	NN	O	O
LTR	NN	O	O
size	NN	O	O
variants	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

One	NN	O	O
HIV-1	NN	O	O
variant	NN	O	O
was	NN	O	O
detected	NN	O	O
containing	NN	O	O
an	NN	O	O
additional	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
Sp1	NN	O	O
.	NN	O	O

Another	NN	O	O
LTR	NN	O	B-DNA
size	NN	O	I-DNA
variation	NN	O	I-DNA
was	NN	O	O
observed	NN	O	O
in	NN	O	O
four	NN	O	O
patients	NN	O	O
in	NN	O	O
a	NN	O	O
region	NN	O	O
just	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
variation	NN	O	O
was	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
a	NN	O	O
duplication	NN	O	O
of	NN	O	O
a	NN	O	O
short	NN	O	B-DNA
DNA	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
CTG-motif	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Cell	NN	O	O
culture	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
natural	NN	O	O
variant	NN	O	O
with	NN	O	O
four	NN	O	O
Sp1	NN	O	B-DNA
sites	NN	O	I-DNA
had	NN	O	O
a	NN	O	O
slightly	NN	O	O
higher	NN	O	O
promoter	NN	O	O
activity	NN	O	O
and	NN	O	O
viral	NN	O	O
replication	NN	O	O
rate	NN	O	O
than	NN	O	O
the	NN	O	O
isogenic	NN	O	B-DNA
control	NN	O	I-DNA
LTR	NN	O	I-DNA
with	NN	O	O
three	NN	O	O
Sp1	NN	O	B-DNA
sites	NN	O	I-DNA
.	NN	O	O

No	NN	O	O
positive	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
duplicated	NN	O	O
CTG-motif	NN	O	B-DNA
could	NN	O	O
be	NN	O	O
detected	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
measure	NN	O	O
small	NN	O	O
differences	NN	O	O
in	NN	O	O
virus	NN	O	O
production	NN	O	O
more	NN	O	O
accurately	NN	O	O
,	NN	O	O
equal	NN	O	O
amounts	NN	O	O
of	NN	O	O
a	NN	O	O
size	NN	O	O
variant	NN	O	O
and	NN	O	O
the	NN	O	O
wild-type	NN	O	B-DNA
plasmid	NN	O	I-DNA
were	NN	O	O
cotransfected	NN	O	O
into	NN	O	O
T-cells	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
virus	NN	O	O
with	NN	O	O
four	NN	O	O
Sp1	NN	O	O
sites	NN	O	O
did	NN	O	O
outgrow	NN	O	O
the	NN	O	O
three	NN	O	O
Sp1	NN	O	O
virus	NN	O	O
in	NN	O	O
35	NN	O	O
days	NN	O	O
of	NN	O	O
culture	NN	O	O
and	NN	O	O
CTG-monomer	NN	O	O
virus	NN	O	O
outcompeted	NN	O	O
the	NN	O	O
CTG-dimer	NN	O	O
virus	NN	O	O
in	NN	O	O
42	NN	O	O
days	NN	O	O
.	NN	O	O

Based	NN	O	O
on	NN	O	O
these	NN	O	O
results	NN	O	O
we	NN	O	O
estimate	NN	O	O
a	NN	O	O
5-10	NN	O	O
%	NN	O	O
difference	NN	O	O
in	NN	O	O
virus	NN	O	O
production	NN	O	O
of	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
variants	NN	O	I-DNA
when	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
wild-type	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	B-protein
B	NN	O	I-protein
cell-derived	NN	O	I-protein
coactivator	NN	O	I-protein
potentiates	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
promoters	NN	O	I-DNA
by	NN	O	O
octamer-binding	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

A	NN	O	O
novel	NN	O	O
B	NN	O	O
cell-restricted	NN	O	O
activity	NN	O	O
,	NN	O	O
required	NN	O	O
for	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
octamer	NN	O	B-DNA
/	NN	O	O
Oct	NN	O	B-DNA
-dependent	NN	O	O
transcription	NN	O	O
from	NN	O	O
an	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
(	NN	O	I-DNA
IgH	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
system	NN	O	O
consisting	NN	O	O
of	NN	O	O
HeLa	NN	O	B-cell_line
cell	NN	O	I-cell_line
-derived	NN	O	O
extracts	NN	O	O
complemented	NN	O	O
with	NN	O	O
fractionated	NN	O	B-protein
B	NN	O	I-protein
cell	NN	O	I-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
factor	NN	O	O
responsible	NN	O	O
for	NN	O	O
this	NN	O	O
activity	NN	O	O
was	NN	O	O
designated	NN	O	O
Oct	NN	O	B-protein
coactivator	NN	O	I-protein
from	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
OCA-B	NN	O	B-protein
)	NN	O	O
.	NN	O	O

OCA-B	NN	O	B-protein
stimulates	NN	O	O
the	NN	O	O
transcription	NN	O	O
from	NN	O	O
an	NN	O	O
IgH	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
either	NN	O	O
Oct-1	NN	O	B-protein
or	NN	O	O
Oct-2	NN	O	B-protein
but	NN	O	O
shows	NN	O	O
no	NN	O	O
significant	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
/	NN	O	O
Oct	NN	O	B-DNA
-dependent	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
ubiquitously	NN	O	B-DNA
expressed	NN	O	I-DNA
histone	NN	O	I-DNA
H2B	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
USF-	NN	O	B-DNA
and	NN	O	I-DNA
Sp1-regulated	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
OCA-B	NN	O	B-protein
is	NN	O	O
a	NN	O	O
tissue-	NN	O	B-protein
,	NN	O	I-protein
promoter-	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
factor-specific	NN	O	I-protein
coactivator	NN	O	I-protein
and	NN	O	O
that	NN	O	O
OCA-B	NN	O	B-protein
may	NN	O	O
be	NN	O	O
a	NN	O	O
major	NN	O	O
determinant	NN	O	O
for	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
activation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
light	NN	O	O
of	NN	O	O
the	NN	O	O
evidence	NN	O	O
showing	NN	O	O
physical	NN	O	O
and	NN	O	O
functional	NN	O	O
interactions	NN	O	O
between	NN	O	O
Oct	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
OCA-B	NN	O	B-protein
,	NN	O	O
we	NN	O	O
propose	NN	O	O
a	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
for	NN	O	O
OCA-B	NN	O	B-protein
and	NN	O	O
discuss	NN	O	O
the	NN	O	O
implications	NN	O	O
of	NN	O	O
OCA-B	NN	O	B-protein
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
other	NN	O	O
tissue-specific	NN	O	B-DNA
promoters	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

SCL	NN	O	B-protein
and	NN	O	O
related	NN	O	O
hemopoietic	NN	O	B-protein
helix-loop-helix	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
helix-loop-helix	NN	O	B-protein
(	NN	O	I-protein
HLH	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
include	NN	O	O
proteins	NN	O	O
critical	NN	O	O
to	NN	O	O
differentiation	NN	O	O
and	NN	O	O
development	NN	O	O
in	NN	O	O
species	NN	O	O
ranging	NN	O	O
from	NN	O	O
plants	NN	O	O
to	NN	O	O
mammals	NN	O	O
.	NN	O	O

Five	NN	O	O
members	NN	O	O
of	NN	O	O
this	NN	O	O
family	NN	O	O
(	NN	O	O
MYC	NN	O	B-protein
,	NN	O	O
SCL	NN	O	B-protein
,	NN	O	O
TAL-2	NN	O	B-protein
,	NN	O	O
LYL-1	NN	O	B-protein
and	NN	O	O
E2A	NN	O	B-protein
)	NN	O	O
are	NN	O	O
implicated	NN	O	O
in	NN	O	O
oncogenic	NN	O	O
events	NN	O	O
in	NN	O	O
human	NN	O	O
lymphoid	NN	O	O
tumors	NN	O	O
because	NN	O	O
of	NN	O	O
their	NN	O	O
consistent	NN	O	O
involvement	NN	O	O
in	NN	O	O
chromosomal	NN	O	O
translocations	NN	O	O
.	NN	O	O

Although	NN	O	O
activated	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemias	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
SCL	NN	O	B-protein
and	NN	O	O
LYL-1	NN	O	B-protein
is	NN	O	O
low	NN	O	O
or	NN	O	O
undetectable	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
populations	NN	O	I-cell_type
.	NN	O	O

SCL	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
,	NN	O	I-cell_type
megakaryocyte	NN	O	I-cell_type
and	NN	O	I-cell_type
mast	NN	O	I-cell_type
cell	NN	O	I-cell_type
populations	NN	O	I-cell_type
(	NN	O	O
the	NN	O	O
same	NN	O	O
cell	NN	O	O
lineages	NN	O	O
as	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
zinc-finger	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
both	NN	O	O
SCL	NN	O	B-protein
and	NN	O	O
GATA-1	NN	O	B-protein
undergo	NN	O	O
coordinate	NN	O	O
modulation	NN	O	O
during	NN	O	O
chemically	NN	O	O
induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
mouse	NN	O	B-cell_type
erythroleukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
are	NN	O	O
down-modulated	NN	O	O
during	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
K562	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
thus	NN	O	O
implying	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
SCL	NN	O	B-protein
in	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
events	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
GATA-1	NN	O	B-protein
,	NN	O	O
SCL	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
developing	NN	O	O
brain	NN	O	O
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
SCL	NN	O	B-protein
suggest	NN	O	O
it	NN	O	O
is	NN	O	O
also	NN	O	O
important	NN	O	O
in	NN	O	O
proliferation	NN	O	O
and	NN	O	O
self-renewal	NN	O	O
events	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
the	NN	O	O
POU	NN	O	B-protein
domain	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Oct-2	NN	O	B-protein
during	NN	O	O
T-cell	NN	O	B-cell_type
activation	NN	O	O
by	NN	O	O
cognate	NN	O	O
antigen	NN	O	O
.	NN	O	O

Oct-2	NN	O	B-protein
is	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
specifically	NN	O	O
to	NN	O	O
octamer	NN	O	B-DNA
DNA	NN	O	I-DNA
motifs	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
immunoglobulin	NN	O	B-DNA
and	NN	O	I-DNA
interleukin-2	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

All	NN	O	O
tumor	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
B-cell	NN	O	B-cell_type
lineage	NN	O	I-cell_type
and	NN	O	O
a	NN	O	O
few	NN	O	O
from	NN	O	O
the	NN	O	O
T-cell	NN	O	B-cell_type
lineage	NN	O	I-cell_type
express	NN	O	O
Oct-2	NN	O	B-protein
.	NN	O	O

To	NN	O	O
address	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
Oct-2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
T-cell	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Oct-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
in	NN	O	O
nontransformed	NN	O	B-cell_line
human	NN	O	I-cell_line
and	NN	O	I-cell_line
mouse	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Oct-2	NN	O	B-protein
was	NN	O	O
found	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_line
and	NN	O	I-cell_line
CD8+	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
prepared	NN	O	O
from	NN	O	O
human	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
and	NN	O	O
in	NN	O	O
mouse	NN	O	B-cell_type
lymph	NN	O	I-cell_type
node	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
a	NN	O	O
T-cell	NN	O	B-cell_line
clone	NN	O	I-cell_line
specific	NN	O	O
for	NN	O	O
pigeon	NN	O	B-protein
cytochrome	NN	O	I-protein
c	NN	O	I-protein
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
I-Ek	NN	O	B-protein
,	NN	O	O
Oct-2	NN	O	B-protein
was	NN	O	O
induced	NN	O	O
by	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
Oct-2	NN	O	B-protein
protein	NN	O	I-protein
seen	NN	O	O
first	NN	O	O
at	NN	O	O
3	NN	O	O
h	NN	O	O
after	NN	O	O
activation	NN	O	O
and	NN	O	O
continuing	NN	O	O
for	NN	O	O
at	NN	O	O
least	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

Oct-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
during	NN	O	O
antigen-driven	NN	O	O
T-cell	NN	O	B-cell_type
activation	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
Oct-2	NN	O	B-protein
participates	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
during	NN	O	O
T-cell	NN	O	B-cell_type
activation.	NN	O	O
The	NN	O	O
relatively	NN	O	O
delayed	NN	O	O
kinetics	NN	O	O
of	NN	O	O
Oct-2	NN	O	B-protein
induction	NN	O	O
suggests	NN	O	O
that	NN	O	O
Oct-2	NN	O	B-protein
mediates	NN	O	O
the	NN	O	O
changes	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
which	NN	O	O
occur	NN	O	O
many	NN	O	O
hours	NN	O	O
or	NN	O	O
days	NN	O	O
following	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
patients	NN	O	O
with	NN	O	O
lupus	NN	O	O
nephritis	NN	O	O
:	NN	O	O
relationship	NN	O	O
between	NN	O	O
receptor	NN	O	O
levels	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
and	NN	O	O
effect	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
clinical	NN	O	O
significance	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
determination	NN	O	O
in	NN	O	O
20	NN	O	O
patients	NN	O	O
with	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
(	NN	O	O
SLE	NN	O	O
)	NN	O	O
who	NN	O	O
afterwards	NN	O	O
developed	NN	O	O
nephrotic	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentrations	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
MNL	NN	O	B-cell_type
)	NN	O	O
in	NN	O	O
these	NN	O	O
patients	NN	O	O
were	NN	O	O
comparable	NN	O	O
with	NN	O	O
those	NN	O	O
in	NN	O	O
both	NN	O	O
other	NN	O	O
patients	NN	O	O
with	NN	O	O
SLE	NN	O	O
and	NN	O	O
healthy	NN	O	O
persons	NN	O	O
.	NN	O	O

Improvement	NN	O	O
in	NN	O	O
urinary	NN	O	O
protein	NN	O	O
excretion	NN	O	O
and	NN	O	O
in	NN	O	O
disease	NN	O	O
activity	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
scored	NN	O	O
according	NN	O	O
to	NN	O	O
the	NN	O	O
SLE	NN	O	O
Disease	NN	O	O
Activity	NN	O	O
Index	NN	O	O
system	NN	O	O
of	NN	O	O
the	NN	O	O
University	NN	O	O
of	NN	O	O
Toronto	NN	O	O
,	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentrations	NN	O	O
in	NN	O	O
MNL	NN	O	B-cell_type
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
corresponding	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
determination	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
lupus	NN	O	O
nephritis	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
predictive	NN	O	O
clue	NN	O	O
for	NN	O	O
assessing	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Changes	NN	O	O
in	NN	O	O
plasma	NN	O	B-protein
interleukin-1	NN	O	I-protein
and	NN	O	O
their	NN	O	O
possible	NN	O	O
relationship	NN	O	O
with	NN	O	O
the	NN	O	O
changes	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
aged	NN	O	O
long-distance	NN	O	O
runner	NN	O	O
]	NN	O	O

For	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
the	NN	O	O
changes	NN	O	O
in	NN	O	O
plasma	NN	O	B-protein
interleukin-1	NN	O	I-protein
(	NN	O	O
IL-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
their	NN	O	O
possible	NN	O	O
relationship	NN	O	O
with	NN	O	O
the	NN	O	O
changes	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
plasma	NN	O	B-protein
IL-1	NN	O	I-protein
and	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
aged	NN	O	O
long-distance	NN	O	O
runner	NN	O	O
were	NN	O	O
measured	NN	O	O
simultaneously	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
IL-1	NN	O	B-protein
was	NN	O	O
expressed	NN	O	O
as	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
stimulate	NN	O	O
3H-TdR	NN	O	B-protein
incorporation	NN	O	O
in	NN	O	O
the	NN	O	O
thymocytes	NN	O	B-cell_type
of	NN	O	O
C57	NN	O	O
mice	NN	O	O
.	NN	O	O

GR	NN	O	B-protein
was	NN	O	O
determined	NN	O	O
by	NN	O	O
whole	NN	O	O
cell	NN	O	O
assay	NN	O	O
with	NN	O	O
3H-Dex	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
plasma	NN	O	O
IL-1	NN	O	B-protein
in	NN	O	O
aged	NN	O	O
long-distance	NN	O	O
runner	NN	O	O
was	NN	O	O
209	NN	O	O
%	NN	O	O
,	NN	O	O
223	NN	O	O
%	NN	O	O
and	NN	O	O
145	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
control	NN	O	O
at	NN	O	O
14.7-18.7	NN	O	O
,	NN	O	O
3.8-7.0	NN	O	O
and	NN	O	O
1.5-2.6	NN	O	O
KD	NN	O	O
fractions	NN	O	O
.	NN	O	O

The	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
aged	NN	O	O
runner	NN	O	O
was	NN	O	O
65	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
control	NN	O	O
.	NN	O	O

Possible	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
changes	NN	O	O
in	NN	O	O
IL-1	NN	O	B-protein
and	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
aged	NN	O	O
long-distance	NN	O	O
runner	NN	O	O
and	NN	O	O
its	NN	O	O
physiological	NN	O	O
significance	NN	O	O
are	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mineralocorticoid	NN	O	O
effector	NN	O	O
mechanism	NN	O	O
in	NN	O	O
preeclampsia	NN	O	O
.	NN	O	O

Mineralocorticoid	NN	O	O
effector	NN	O	O
mechanisms	NN	O	O
were	NN	O	O
evaluated	NN	O	O
in	NN	O	O
29	NN	O	O
patients	NN	O	O
with	NN	O	O
preeclampsia	NN	O	O
and	NN	O	O
in	NN	O	O
25	NN	O	O
uncomplicated	NN	O	O
pregnancies	NN	O	O
by	NN	O	O
measurement	NN	O	O
of	NN	O	O
plasma	NN	O	O
aldosterone	NN	O	O
,	NN	O	O
levels	NN	O	O
of	NN	O	O
mineralocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
MR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leucocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
subtraction	NN	O	O
potential	NN	O	O
difference	NN	O	O
(	NN	O	O
SPD	NN	O	O
;	NN	O	O
rectal	NN	O	O
minus	NN	O	O
oral	NN	O	O
values	NN	O	O
)	NN	O	O
.	NN	O	O

Mean	NN	O	O
values	NN	O	O
for	NN	O	O
plasma	NN	O	O
aldosterone	NN	O	O
were	NN	O	O
not	NN	O	O
different	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
groups	NN	O	O
,	NN	O	O
but	NN	O	O
significant	NN	O	O
differences	NN	O	O
were	NN	O	O
observed	NN	O	O
for	NN	O	O
MR	NN	O	B-protein
(	NN	O	O
preeclampsia	NN	O	O
,	NN	O	O
81	NN	O	O
+/-	NN	O	O
44	NN	O	O
receptors/cell	NN	O	O
;	NN	O	O
controls	NN	O	O
,	NN	O	O
306	NN	O	O
+/-	NN	O	O
168	NN	O	O
)	NN	O	O
and	NN	O	O
SPD	NN	O	O
(	NN	O	O
preeclampsia	NN	O	O
,	NN	O	O
65	NN	O	O
+/-	NN	O	O
7	NN	O	O
mV	NN	O	O
;	NN	O	O
controls	NN	O	O
,	NN	O	O
12	NN	O	O
+/-	NN	O	O
5	NN	O	O
mV	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
six	NN	O	O
cases	NN	O	O
we	NN	O	O
determined	NN	O	O
MR	NN	O	B-protein
,	NN	O	O
plasma	NN	O	O
aldosterone	NN	O	O
,	NN	O	O
and	NN	O	O
SPD	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
preeclampsia	NN	O	O
before	NN	O	O
and	NN	O	O
3	NN	O	O
months	NN	O	O
after	NN	O	O
delivery	NN	O	O
.	NN	O	O

MR	NN	O	B-protein
were	NN	O	O
reduced	NN	O	O
before	NN	O	O
delivery	NN	O	O
(	NN	O	O
96	NN	O	O
+/-	NN	O	O
27	NN	O	O
receptors/cell	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
SPD	NN	O	O
increased	NN	O	O
(	NN	O	O
64	NN	O	O
+/-	NN	O	O
8	NN	O	O
mV	NN	O	O
)	NN	O	O
,	NN	O	O
with	NN	O	O
both	NN	O	O
parameters	NN	O	O
normalizing	NN	O	O
after	NN	O	O
delivery	NN	O	O
(	NN	O	O
MR	NN	O	B-protein
,	NN	O	O
242	NN	O	O
+/-	NN	O	O
79	NN	O	O
;	NN	O	O
SPD	NN	O	O
,	NN	O	O
14.0	NN	O	O
+/-	NN	O	O
4	NN	O	O
mV	NN	O	O
)	NN	O	O
.	NN	O	O

Aldosterone	NN	O	O
levels	NN	O	O
returned	NN	O	O
to	NN	O	O
normal	NN	O	O
nonpregnant	NN	O	O
values	NN	O	O
after	NN	O	O
delivery	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
for	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
mineralocorticoid	NN	O	O
effector	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
the	NN	O	O
etiology	NN	O	O
of	NN	O	O
preeclampsia	NN	O	O
and	NN	O	O
could	NN	O	O
be	NN	O	O
an	NN	O	O
useful	NN	O	O
marker	NN	O	O
for	NN	O	O
diagnosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Kinetics	NN	O	O
of	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
turnover	NN	O	O
of	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_line
HL60	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
--	NN	O	O
coincident	NN	O	O
rise	NN	O	O
of	NN	O	O
DNA-relaxing	NN	O	O
activity	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
.	NN	O	O

High	NN	O	B-protein
affinity	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
for	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxycholecalciferol	NN	O	O
(	NN	O	O
calcitriol	NN	O	O
)	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
HL60	NN	O	O
human	NN	O	O
leukemia	NN	O	O
cells	NN	O	O
and	NN	O	O
in	NN	O	O
low	NN	O	O
numbers	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
PBL	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
expressing	NN	O	O
some	NN	O	O
characteristics	NN	O	O
of	NN	O	O
promyelocytes	NN	O	B-cell_line
,	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
to	NN	O	O
monocytoid	NN	O	O
differentiation	NN	O	O
by	NN	O	O
calcitriol	NN	O	O
.	NN	O	O

Specific	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
[	NN	O	B-protein
3H	NN	O	I-protein
]	NN	O	I-protein
calcitriol/VDR	NN	O	I-protein
was	NN	O	O
examined	NN	O	O
after	NN	O	O
exposure	NN	O	O
of	NN	O	O
whole	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
M/l	NN	O	O
calcitriol	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
and	NN	O	O
absence	NN	O	O
of	NN	O	O
a	NN	O	O
500-fold	NN	O	O
excess	NN	O	O
of	NN	O	O
unlabeled	NN	O	O
ligand	NN	O	O
and	NN	O	O
subsequent	NN	O	O
isolation	NN	O	O
of	NN	O	O
nuclei	NN	O	O
.	NN	O	O

Specific	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
[	NN	O	B-protein
3H	NN	O	I-protein
]	NN	O	I-protein
calcitriol/VDR	NN	O	I-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
time	NN	O	O
dependent	NN	O	O
reaching	NN	O	O
a	NN	O	O
maximum	NN	O	O
of	NN	O	O
approximately	NN	O	O
2100	NN	O	O
binding	NN	O	O
sites/nucleus	NN	O	O
after	NN	O	O
3	NN	O	O
h	NN	O	O
of	NN	O	O
incubation	NN	O	O
in	NN	O	O
HL60	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
maximum	NN	O	O
of	NN	O	O
approximately	NN	O	O
310	NN	O	O
binding	NN	O	O
sites/nucleus	NN	O	O
was	NN	O	O
found	NN	O	O
after	NN	O	O
3	NN	O	O
h	NN	O	O
in	NN	O	O
PBL	NN	O	B-cell_type
.	NN	O	O

Pulse	NN	O	O
exposure	NN	O	O
of	NN	O	O
HL60	NN	O	B-cell_line
to	NN	O	O
radiolabeled	NN	O	O
hormone	NN	O	O
for	NN	O	O
3	NN	O	O
h	NN	O	O
followed	NN	O	O
by	NN	O	O
culture	NN	O	O
in	NN	O	O
medium	NN	O	O
without	NN	O	O
serum	NN	O	O
and	NN	O	O
calcitriol	NN	O	O
lead	NN	O	O
to	NN	O	O
nuclear	NN	O	O
retention	NN	O	O
of	NN	O	O
approximately	NN	O	O
1600	NN	O	B-protein
radiolabeled	NN	O	I-protein
VDR	NN	O	I-protein
by	NN	O	O
8	NN	O	O
h	NN	O	O
and	NN	O	O
approximately	NN	O	O
1000	NN	O	O
VDR	NN	O	B-protein
by	NN	O	O
24	NN	O	O
h	NN	O	O
.	NN	O	O

Radiolabeled	NN	O	B-protein
VDR	NN	O	I-protein
disappeared	NN	O	O
from	NN	O	O
the	NN	O	O
nuclear	NN	O	O
compartment	NN	O	O
with	NN	O	O
a	NN	O	O
halflife	NN	O	O
of	NN	O	O
approximately	NN	O	O
30	NN	O	O
min	NN	O	O
if	NN	O	O
cells	NN	O	O
were	NN	O	O
cultured	NN	O	O
with	NN	O	O
identical	NN	O	O
concentrations	NN	O	O
of	NN	O	O
unlabeled	NN	O	O
hormone	NN	O	O
after	NN	O	O
the	NN	O	O
pulse	NN	O	O
(	NN	O	O
pulse/chase-experiments	NN	O	O
)	NN	O	O
.	NN	O	O

No	NN	O	O
difference	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
retention	NN	O	O
in	NN	O	O
pulse	NN	O	O
and	NN	O	O
pulse/chase-experiments	NN	O	O
was	NN	O	O
seen	NN	O	O
in	NN	O	O
PBL	NN	O	B-cell_type
,	NN	O	O
where	NN	O	O
VDR	NN	O	B-protein
halflife	NN	O	O
was	NN	O	O
approximately	NN	O	O
30	NN	O	O
min	NN	O	O
.	NN	O	O

No	NN	O	O
specific	NN	O	O
translocation	NN	O	O
into	NN	O	O
the	NN	O	O
nuclear	NN	O	O
compartment	NN	O	O
was	NN	O	O
seen	NN	O	O
when	NN	O	O
isolated	NN	O	O
nuclei	NN	O	O
were	NN	O	O
incubated	NN	O	O
in	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
calcitriol	NN	O	O
.	NN	O	O

Radiolabeled	NN	O	B-protein
hormone/receptor	NN	O	I-protein
complexes	NN	O	I-protein
of	NN	O	O
nuclei	NN	O	O
isolated	NN	O	O
from	NN	O	O
cells	NN	O	O
exposed	NN	O	O
for	NN	O	O
3	NN	O	O
h	NN	O	O
to	NN	O	O
radiolabeled	NN	O	O
hormone	NN	O	O
--	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
identical	NN	O	O
experiments	NN	O	O
with	NN	O	O
intact	NN	O	O
cells	NN	O	O
--	NN	O	O
did	NN	O	O
not	NN	O	O
disappear	NN	O	O
from	NN	O	O
the	NN	O	O
nuclear	NN	O	O
compartment	NN	O	O
upon	NN	O	O
incubation	NN	O	O
of	NN	O	O
nuclei	NN	O	O
with	NN	O	O
identical	NN	O	O
concentrations	NN	O	O
of	NN	O	O
the	NN	O	O
unlabeled	NN	O	O
compound	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
DNA	NN	O	B-protein
relaxing	NN	O	I-protein
enzymes	NN	O	I-protein
(	NN	O	O
e.g	NN	O	O
.	NN	O	O
topoisomerases	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
II	NN	O	I-protein
)	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
was	NN	O	O
measured	NN	O	O
using	NN	O	O
a	NN	O	O
PBR	NN	O	O
322-relaxation-assay	NN	O	O
.	NN	O	O

Enhanced	NN	O	O
overall	NN	O	O
enzyme	NN	O	O
activity	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
by	NN	O	O
1	NN	O	O
h	NN	O	O
after	NN	O	O
incubation	NN	O	O
with	NN	O	O
calcitriol	NN	O	O
(	NN	O	O
final	NN	O	O
ethanol	NN	O	O
concentration	NN	O	O
0.0001	NN	O	O
%	NN	O	O
v/v	NN	O	O
)	NN	O	O
in	NN	O	O
HL60	NN	O	B-cell_line
and	NN	O	O
PBL	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
enhanced	NN	O	O
activity	NN	O	O
disappeared	NN	O	O
after	NN	O	O
2	NN	O	O
h	NN	O	O
in	NN	O	O
PBL	NN	O	B-cell_type
,	NN	O	O
whereas	NN	O	O
it	NN	O	O
was	NN	O	O
still	NN	O	O
enhanced	NN	O	O
by	NN	O	O
4	NN	O	O
h	NN	O	O
in	NN	O	O
HL60	NN	O	B-cell_line
.	NN	O	O

No	NN	O	O
effect	NN	O	O
was	NN	O	O
seen	NN	O	O
in	NN	O	O
ethanol	NN	O	B-cell_line
treated	NN	O	I-cell_line
controls	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
a	NN	O	O
specific	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
mechanism	NN	O	O
exists	NN	O	O
for	NN	O	O
calcitriol	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
examined	NN	O	O
,	NN	O	O
most	NN	O	O
likely	NN	O	O
due	NN	O	O
to	NN	O	O
translocation	NN	O	O
of	NN	O	O
receptor	NN	O	B-protein
proteins	NN	O	I-protein
after	NN	O	O
hormone	NN	O	O
binding	NN	O	O
.	NN	O	O

Translocated	NN	O	B-protein
hormone/receptor	NN	O	I-protein
complexes	NN	O	I-protein
compete	NN	O	O
for	NN	O	O
a	NN	O	O
limited	NN	O	O
number	NN	O	O
of	NN	O	O
specific	NN	O	O
nuclear	NN	O	O
binding	NN	O	O
sites	NN	O	O
.	NN	O	O

Enhanced	NN	O	O
activity	NN	O	O
of	NN	O	O
topoisomerases	NN	O	B-protein
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
upon	NN	O	O
translocation	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
might	NN	O	O
reflect	NN	O	O
interaction	NN	O	O
of	NN	O	O
both	NN	O	O
within	NN	O	O
the	NN	O	O
nuclear	NN	O	O
compartment	NN	O	O
,	NN	O	O
thus	NN	O	O
initiating	NN	O	O
DNA-unwinding	NN	O	O
,	NN	O	O
a	NN	O	O
prerequisite	NN	O	O
of	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Influence	NN	O	O
of	NN	O	O
estradiol	NN	O	O
and	NN	O	O
tamoxifen	NN	O	O
on	NN	O	O
susceptibility	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
breast	NN	O	I-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
to	NN	O	O
lysis	NN	O	O
by	NN	O	O
lymphokine-activated	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
design	NN	O	O
of	NN	O	O
combination	NN	O	O
hormonal	NN	O	O
and	NN	O	O
immunotherapeutic	NN	O	O
protocols	NN	O	O
for	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
may	NN	O	O
be	NN	O	O
facilitated	NN	O	O
by	NN	O	O
analysis	NN	O	O
of	NN	O	O
preclinical	NN	O	O
in	NN	O	O
vitro	NN	O	O
model	NN	O	O
systems	NN	O	O
.	NN	O	O

Estrogen	NN	O	B-cell_line
receptor	NN	O	I-cell_line
positive	NN	O	I-cell_line
(	NN	O	I-cell_line
ER+	NN	O	I-cell_line
:	NN	O	I-cell_line
MCF-7	NN	O	I-cell_line
)	NN	O	I-cell_line
and	NN	O	I-cell_line
negative	NN	O	I-cell_line
(	NN	O	I-cell_line
ER-	NN	O	I-cell_line
:	NN	O	I-cell_line
MDA-MB-231	NN	O	I-cell_line
)	NN	O	I-cell_line
human	NN	O	I-cell_line
breast	NN	O	I-cell_line
cancer	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
utilized	NN	O	O
to	NN	O	O
evaluate	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
tamoxifen	NN	O	O
(	NN	O	O
TAM	NN	O	O
)	NN	O	O
and	NN	O	O
estradiol	NN	O	O
(	NN	O	O
E2	NN	O	O
)	NN	O	O
on	NN	O	O
modulation	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
target	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
lysis	NN	O	O
by	NN	O	O
lymphokine-activated	NN	O	B-cell_type
killer	NN	O	I-cell_type
(	NN	O	I-cell_type
LAK	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

E2-stimulated	NN	O	B-cell_line
ER+	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
more	NN	O	O
susceptible	NN	O	O
to	NN	O	O
lysis	NN	O	O
by	NN	O	O
LAK	NN	O	B-cell_type
cells	NN	O	I-cell_type
than	NN	O	O
corresponding	NN	O	O
TAM-treated	NN	O	B-cell_line
or	NN	O	O
control	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
treatment	NN	O	O
of	NN	O	O
ER-	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
either	NN	O	O
E2	NN	O	O
or	NN	O	O
TAM	NN	O	O
alone	NN	O	O
did	NN	O	O
not	NN	O	O
alter	NN	O	O
from	NN	O	O
control	NN	O	O
their	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
this	NN	O	O
immune-mediated	NN	O	O
lysis	NN	O	O
.	NN	O	O

All	NN	O	O
ER+	NN	O	B-cell_line
and	NN	O	I-cell_line
ER-	NN	O	I-cell_line
cells	NN	O	I-cell_line
tested	NN	O	O
remained	NN	O	O
sensitive	NN	O	O
after	NN	O	O
treatment	NN	O	O
with	NN	O	O
TAM	NN	O	O
to	NN	O	O
lysis	NN	O	O
by	NN	O	O
LAK	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
an	NN	O	O
adenocarcinoma	NN	O	B-protein
reactive	NN	O	I-protein
human-mouse	NN	O	I-protein
chimeric	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
ING-1	NN	O	B-protein
)	NN	O	O
was	NN	O	O
able	NN	O	O
to	NN	O	O
significantly	NN	O	O
boost	NN	O	O
in	NN	O	O
vivo	NN	O	O
generated	NN	O	O
LAK	NN	O	O
cell-mediated	NN	O	O
lysis	NN	O	O
of	NN	O	O
control	NN	O	B-cell_line
,	NN	O	I-cell_line
E2-treated	NN	O	I-cell_line
,	NN	O	I-cell_line
and	NN	O	I-cell_line
TAM-treated	NN	O	I-cell_line
ER+	NN	O	I-cell_line
and	NN	O	I-cell_line
ER-	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
in	NN	O	O
vitro	NN	O	O
results	NN	O	O
provide	NN	O	O
a	NN	O	O
preclinical	NN	O	O
rationale	NN	O	O
for	NN	O	O
in	NN	O	O
vivo	NN	O	O
testing	NN	O	O
of	NN	O	O
TAM	NN	O	O
,	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
breast	NN	O	O
cancer	NN	O	O
reactive	NN	O	O
antibody-dependent	NN	O	O
cellular	NN	O	O
cytotoxicity	NN	O	O
facilitating	NN	O	O
antibody	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
refractory	NN	O	O
or	NN	O	O
high	NN	O	O
risk	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
cellular	NN	O	B-DNA
oncogene	NN	O	I-DNA
c-myb	NN	O	B-DNA
can	NN	O	O
interact	NN	O	O
synergistically	NN	O	O
with	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
BZLF1	NN	O	I-protein
transactivator	NN	O	I-protein
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Regulation	NN	O	O
of	NN	O	O
replicative	NN	O	O
functions	NN	O	O
in	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
EBV	NN	O	I-DNA
)	NN	O	I-DNA
genome	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
virally	NN	O	B-protein
encoded	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
Z	NN	O	B-protein
(	NN	O	O
BZLF1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
Z	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
AP-1	NN	O	B-DNA
sites	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
homodimer	NN	O	O
and	NN	O	O
has	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
to	NN	O	O
c-Fos	NN	O	B-DNA
,	NN	O	O
can	NN	O	O
efficiently	NN	O	O
activate	NN	O	O
the	NN	O	O
EBV	NN	O	B-DNA
early	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
BMRF1	NN	O	B-DNA
,	NN	O	O
in	NN	O	O
certain	NN	O	O
cell	NN	O	O
types	NN	O	O
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
but	NN	O	O
not	NN	O	O
others	NN	O	O
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
c-myb	NN	O	B-protein
proto-oncogene	NN	O	I-protein
product	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
itself	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
transcriptional	NN	O	B-protein
transactivator	NN	O	I-protein
,	NN	O	O
can	NN	O	O
interact	NN	O	O
synergistically	NN	O	O
with	NN	O	O
Z	NN	O	B-protein
in	NN	O	O
activating	NN	O	O
the	NN	O	O
BMRF1	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
a	NN	O	O
T-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
)	NN	O	O
or	NN	O	O
Raji	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
an	NN	O	O
EBV-positive	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
c-myb	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
by	NN	O	O
itself	NN	O	O
has	NN	O	O
little	NN	O	O
effect	NN	O	O
.	NN	O	O

The	NN	O	O
simian	NN	O	B-DNA
virus	NN	O	I-DNA
40	NN	O	I-DNA
early	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
also	NN	O	O
synergistically	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
Z/c-myb	NN	O	B-protein
combination	NN	O	I-protein
.	NN	O	O

Synergistic	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
BMRF1	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
Z/c-myb	NN	O	B-protein
combination	NN	O	I-protein
appears	NN	O	O
to	NN	O	O
involve	NN	O	O
direct	NN	O	O
binding	NN	O	O
by	NN	O	O
the	NN	O	O
Z	NN	O	B-protein
protein	NN	O	I-protein
but	NN	O	O
not	NN	O	O
the	NN	O	O
c-myb	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

A	NN	O	O
30-bp	NN	O	B-DNA
sequence	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
BMRF1	NN	O	B-DNA
promoter	NN	O	I-DNA
which	NN	O	O
contains	NN	O	O
a	NN	O	O
Z	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
(	NN	O	O
a	NN	O	O
consensus	NN	O	B-DNA
AP-1	NN	O	I-DNA
site	NN	O	I-DNA
)	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
transfer	NN	O	O
high-level	NN	O	O
lymphoid-specific	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
the	NN	O	O
Z/c-myb	NN	O	B-protein
combination	NN	O	I-protein
to	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

That	NN	O	O
the	NN	O	O
c-myb	NN	O	B-protein
oncogene	NN	O	I-protein
product	NN	O	I-protein
can	NN	O	O
interact	NN	O	O
synergistically	NN	O	O
with	NN	O	O
an	NN	O	O
EBV-encoded	NN	O	B-protein
member	NN	O	I-protein
of	NN	O	O
the	NN	O	O
leucine	NN	O	B-protein
zipper	NN	O	I-protein
protein	NN	O	I-protein
family	NN	O	I-protein
suggests	NN	O	O
c-myb	NN	O	B-DNA
is	NN	O	O
likely	NN	O	O
to	NN	O	O
engage	NN	O	O
in	NN	O	O
similar	NN	O	O
interactions	NN	O	O
with	NN	O	O
cellularly	NN	O	B-protein
encoded	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Alpha-lipoic	NN	O	O
acid	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Acquired	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
(	NN	O	O
AIDS	NN	O	O
)	NN	O	O
results	NN	O	O
from	NN	O	O
infection	NN	O	O
with	NN	O	O
a	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	I-DNA
LTR	NN	O	I-DNA
)	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
HIV	NN	O	B-DNA
proviral	NN	O	I-DNA
DNA	NN	O	I-DNA
contains	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
appears	NN	O	O
to	NN	O	O
regulate	NN	O	O
HIV	NN	O	O
activation	NN	O	O
.	NN	O	O

Recent	NN	O	O
findings	NN	O	O
suggest	NN	O	O
an	NN	O	O
involvement	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
species	NN	O	O
(	NN	O	O
ROS	NN	O	O
)	NN	O	O
in	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
leading	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
was	NN	O	O
based	NN	O	O
on	NN	O	O
reports	NN	O	O
that	NN	O	O
antioxidants	NN	O	O
which	NN	O	O
eliminate	NN	O	O
ROS	NN	O	O
should	NN	O	O
block	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
subsequently	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
thus	NN	O	O
antioxidants	NN	O	O
can	NN	O	O
be	NN	O	O
used	NN	O	O
as	NN	O	O
therapeutic	NN	O	O
agents	NN	O	O
for	NN	O	O
AIDS	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
1	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
cells/ml	NN	O	O
)	NN	O	O
with	NN	O	O
a	NN	O	O
natural	NN	O	O
thiol	NN	O	O
antioxidant	NN	O	O
,	NN	O	O
alpha-lipoic	NN	O	O
acid	NN	O	O
,	NN	O	O
prior	NN	O	O
to	NN	O	O
the	NN	O	O
stimulation	NN	O	O
of	NN	O	O
cells	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
inhibit	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
(	NN	O	O
25	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
or	NN	O	O
by	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
50	NN	O	O
ng/ml	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
action	NN	O	O
of	NN	O	O
alpha-lipoic	NN	O	O
acid	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
very	NN	O	O
potent	NN	O	O
as	NN	O	O
only	NN	O	O
4	NN	O	O
mM	NN	O	O
was	NN	O	O
needed	NN	O	O
for	NN	O	O
a	NN	O	O
complete	NN	O	O
inhibition	NN	O	O
,	NN	O	O
whereas	NN	O	O
20	NN	O	O
mM	NN	O	O
was	NN	O	O
required	NN	O	O
for	NN	O	O
N-acetylcysteine	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
alpha-lipoic	NN	O	O
acid	NN	O	O
may	NN	O	O
be	NN	O	O
effective	NN	O	O
in	NN	O	O
AIDS	NN	O	O
therapeutics	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reticuloendotheliosis	NN	O	B-DNA
virus	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
elements	NN	O	I-DNA
are	NN	O	O
efficient	NN	O	O
promoters	NN	O	B-DNA
in	NN	O	O
cells	NN	O	O
of	NN	O	O
various	NN	O	O
species	NN	O	O
and	NN	O	O
tissue	NN	O	O
origin	NN	O	O
,	NN	O	O
including	NN	O	O
human	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Promiscuous	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
reticuloendotheliosis	NN	O	B-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
REV	NN	O	I-DNA
)	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
transient	NN	O	O
expression	NN	O	O
assays	NN	O	O
using	NN	O	O
LTR-chloramphenicol	NN	O	B-DNA
acetyltransferase-encoding	NN	O	I-DNA
gene	NN	O	I-DNA
chimeras	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
cells	NN	O	O
of	NN	O	O
diverse	NN	O	O
species	NN	O	O
and	NN	O	O
tissue	NN	O	O
type	NN	O	O
;	NN	O	O
levels	NN	O	O
of	NN	O	O
expression	NN	O	O
from	NN	O	O
two	NN	O	O
different	NN	O	O
REV	NN	O	B-DNA
LTRs	NN	O	I-DNA
correlate	NN	O	O
with	NN	O	O
reports	NN	O	O
of	NN	O	O
pathogenicity	NN	O	O
of	NN	O	O
the	NN	O	O
respective	NN	O	O
viruses	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

REVs	NN	O	O
do	NN	O	O
not	NN	O	O
encode	NN	O	O
a	NN	O	O
transactivator	NN	O	O
targeted	NN	O	O
to	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
LTR	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
Marek	NN	O	O
's	NN	O	O
disease	NN	O	O
virus	NN	O	O
,	NN	O	O
a	NN	O	O
herpesvirus	NN	O	O
with	NN	O	O
an	NN	O	O
overlapping	NN	O	O
host	NN	O	O
range	NN	O	O
,	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
factors	NN	O	O
that	NN	O	O
preferentially	NN	O	O
enhance	NN	O	O
expression	NN	O	O
from	NN	O	O
REV	NN	O	O
or	NN	O	O
avian	NN	O	B-DNA
sarcoma/leukemia	NN	O	I-DNA
virus	NN	O	I-DNA
LTRs	NN	O	I-DNA
.	NN	O	O

REV	NN	O	B-DNA
LTRs	NN	O	I-DNA
work	NN	O	O
efficiently	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
are	NN	O	O
viable	NN	O	O
alternatives	NN	O	O
to	NN	O	O
promoters	NN	O	B-DNA
commonly	NN	O	O
used	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
cloned	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

They	NN	O	O
may	NN	O	O
also	NN	O	O
prove	NN	O	O
useful	NN	O	O
in	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
new	NN	O	O
,	NN	O	O
ubiquitous	NN	O	B-protein
cellular	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
Ets-related	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
Elf-1	NN	O	B-protein
,	NN	O	O
binds	NN	O	O
to	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
2	NN	O	I-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
that	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
inducible	NN	O	O
trans	NN	O	O
activation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
and	NN	O	O
HIV-2	NN	O	O
are	NN	O	O
structurally	NN	O	O
related	NN	O	O
retroviruses	NN	O	O
which	NN	O	O
both	NN	O	O
cause	NN	O	O
AIDS	NN	O	O
in	NN	O	O
humans	NN	O	O
.	NN	O	O

Although	NN	O	O
both	NN	O	O
viruses	NN	O	O
establish	NN	O	O
latency	NN	O	O
in	NN	O	O
quiescent	NN	O	O
human-peripheral-blood	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
asymptomatic	NN	O	O
phase	NN	O	O
of	NN	O	O
HIV-2	NN	O	O
infection	NN	O	O
may	NN	O	O
be	NN	O	O
more	NN	O	O
prolonged	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

The	NN	O	O
latent	NN	O	O
phases	NN	O	O
of	NN	O	O
both	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
HIV-2	NN	O	O
infection	NN	O	O
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
disrupted	NN	O	O
by	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
a	NN	O	O
process	NN	O	O
that	NN	O	O
requires	NN	O	O
host	NN	O	O
cell	NN	O	O
transcription	NN	O	O
factors	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
the	NN	O	O
transcription	NN	O	O
factor	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
for	NN	O	O
inducible	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
factors	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
are	NN	O	O
required	NN	O	O
to	NN	O	O
activate	NN	O	O
HIV-2	NN	O	O
transcription	NN	O	O
in	NN	O	O
infected	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
novel	NN	O	O
Ets-related	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
Elf-1	NN	O	B-protein
,	NN	O	O
binds	NN	O	O
specifically	NN	O	O
to	NN	O	O
two	NN	O	B-DNA
purine-rich	NN	O	I-DNA
motifs	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
HIV-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Mutagenesis	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
these	NN	O	O
Elf-1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV-2	NN	O	O
transcription	NN	O	O
following	NN	O	O
T-cell-receptor-mediated	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
Elf-1	NN	O	B-protein
is	NN	O	O
the	NN	O	O
only	NN	O	O
factor	NN	O	O
present	NN	O	O
in	NN	O	O
activated	NN	O	O
T-cell	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
these	NN	O	O
sites	NN	O	O
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Elf-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
T-cell-receptor-mediated	NN	O	O
trans	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-2	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
may	NN	O	O
explain	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
clinical	NN	O	O
spectra	NN	O	O
of	NN	O	O
diseases	NN	O	O
caused	NN	O	O
by	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
HIV-2	NN	O	O
and	NN	O	O
may	NN	O	O
also	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
design	NN	O	O
of	NN	O	O
therapeutic	NN	O	O
approaches	NN	O	O
to	NN	O	O
HIV-2	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Okadaic	NN	O	O
acid	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
inducer	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor-alpha	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
an	NN	O	O
optimal	NN	O	O
concentration	NN	O	O
of	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
phosphatases	NN	O	B-protein
1	NN	O	I-protein
and	NN	O	I-protein
2A	NN	O	I-protein
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
a	NN	O	O
marked	NN	O	O
increase	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
levels	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
octamer	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
Oct-1	NN	O	B-protein
or	NN	O	O
Oct-2	NN	O	B-protein
,	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

Since	NN	O	O
both	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
have	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor-alpha	NN	O	I-DNA
(	NN	O	I-DNA
TNF-alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
we	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
on	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
striking	NN	O	O
increase	NN	O	O
in	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcripts	NN	O	O
within	NN	O	O
1	NN	O	O
h	NN	O	O
of	NN	O	O
stimulation	NN	O	O
and	NN	O	O
large	NN	O	O
amounts	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
were	NN	O	O
released	NN	O	O
into	NN	O	O
the	NN	O	O
culture	NN	O	O
media	NN	O	O
.	NN	O	O

Although	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
provides	NN	O	O
a	NN	O	O
potent	NN	O	O
inductive	NN	O	O
signal	NN	O	O
for	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
it	NN	O	O
did	NN	O	O
not	NN	O	O
induce	NN	O	O
either	NN	O	O
B	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
or	NN	O	O
immunoglobulin	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

-DOCSTART-	O

Heterodimerization	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
proteins	NN	O	I-protein
represents	NN	O	O
multiple	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
rel	NN	O	B-protein
related	NN	O	I-protein
polypeptides	NN	O	I-protein
that	NN	O	O
contribute	NN	O	O
to	NN	O	O
regulation	NN	O	O
of	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
immune	NN	O	O
responsiveness	NN	O	O
and	NN	O	O
inflammation	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
multiple	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
related	NN	O	I-protein
polypeptides	NN	O	I-protein
ranging	NN	O	O
from	NN	O	O
85	NN	O	O
to	NN	O	O
45	NN	O	O
kDa	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
their	NN	O	O
capacity	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
PRDII	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
interferon	NN	O	B-DNA
beta	NN	O	I-DNA
and	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
possess	NN	O	O
distinct	NN	O	O
intrinsic	NN	O	O
DNA	NN	O	O
binding	NN	O	O
affinities	NN	O	O
for	NN	O	O
this	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
form	NN	O	O
multiple	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
homo-	NN	O	I-protein
and	NN	O	I-protein
heterodimer	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
co-renaturation	NN	O	O
experiments	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
using	NN	O	O
DNA	NN	O	B-DNA
templates	NN	O	I-DNA
containing	NN	O	O
two	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
PRDII	NN	O	B-DNA
domain	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
the	NN	O	O
rabbit	NN	O	B-DNA
beta	NN	O	I-DNA
globin	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
purified	NN	O	O
polypeptides	NN	O	O
specifically	NN	O	O
stimulated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
dependent	NN	O	O
transcription	NN	O	O
in	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
reconstitution	NN	O	O
assay	NN	O	O
as	NN	O	O
heterodimers	NN	O	O
but	NN	O	O
not	NN	O	O
as	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	I-protein
.	NN	O	O

These	NN	O	O
experiments	NN	O	O
emphasize	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
dimerization	NN	O	O
as	NN	O	O
a	NN	O	O
distinct	NN	O	O
level	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
that	NN	O	O
may	NN	O	O
permit	NN	O	O
functional	NN	O	O
diversification	NN	O	O
of	NN	O	O
a	NN	O	O
limited	NN	O	O
number	NN	O	O
of	NN	O	O
regulatory	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
minor	NN	O	O
salivary	NN	O	O
glands	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
Sjogren	NN	O	O
's	NN	O	O
syndrome	NN	O	O
.	NN	O	O

c-myc	NN	O	B-DNA
protooncogene	NN	O	I-DNA
is	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
B	NN	O	O
cell	NN	O	O
lymphoid	NN	O	O
malignancies	NN	O	O
and	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
are	NN	O	O
observed	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Sjogren	NN	O	O
's	NN	O	O
syndrome	NN	O	O
(	NN	O	O
SS	NN	O	O
)	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
lymphocytic	NN	O	B-cell_type
infiltrates	NN	O	I-cell_type
of	NN	O	O
exocrine	NN	O	O
glands	NN	O	O
,	NN	O	O
remarkable	NN	O	O
B	NN	O	O
cell	NN	O	O
hyperreactivity	NN	O	O
and	NN	O	O
a	NN	O	O
strong	NN	O	O
predisposition	NN	O	O
to	NN	O	O
B	NN	O	O
cell	NN	O	O
neoplasia	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
c-myc	NN	O	B-RNA
protooncogene	NN	O	I-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
29	NN	O	O
labial	NN	O	O
minor	NN	O	O
salivary	NN	O	O
gland	NN	O	O
biopsies	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
primary	NN	O	O
SS	NN	O	O
and	NN	O	O
15	NN	O	O
controls	NN	O	O
was	NN	O	O
examined	NN	O	O
using	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
histochemistry	NN	O	O
.	NN	O	O

Two	NN	O	O
40mer	NN	O	O
oligonucleotides	NN	O	O
from	NN	O	O
the	NN	O	O
1st	NN	O	B-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
2nd	NN	O	I-DNA
exon	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
c-myc	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
labeled	NN	O	O
with	NN	O	O
35S	NN	O	O
,	NN	O	O
were	NN	O	O
used	NN	O	O
as	NN	O	O
probes	NN	O	O
.	NN	O	O

To	NN	O	O
detect	NN	O	O
the	NN	O	O
origin	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
hybridized	NN	O	O
with	NN	O	O
a	NN	O	O
c-myc	NN	O	B-DNA
probe	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
combined	NN	O	O
immunochemistry	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
histochemistry	NN	O	O
technique	NN	O	O
was	NN	O	O
used	NN	O	O
.	NN	O	O

High	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
was	NN	O	O
detected	NN	O	O
on	NN	O	O
acinar	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

c-myc	NN	O	B-protein
did	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
c-fos	NN	O	B-protein
and	NN	O	O
c-jun	NN	O	B-protein
protein	NN	O	O
expression	NN	O	O
.	NN	O	O

Stronger	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
labial	NN	O	O
salivary	NN	O	O
glands	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
longer	NN	O	O
disease	NN	O	O
duration	NN	O	O
(	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
or	NN	O	O
equal	NN	O	O
to	NN	O	O
0.002	NN	O	O
)	NN	O	O
and	NN	O	O
more	NN	O	O
intense	NN	O	O
T	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
infiltrates	NN	O	I-cell_type
(	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0.05	NN	O	O
)	NN	O	O
although	NN	O	O
these	NN	O	O
patients	NN	O	O
revealed	NN	O	O
no	NN	O	O
hypergammaglobulinemia	NN	O	O
.	NN	O	O

No	NN	O	O
correlation	NN	O	O
was	NN	O	O
observed	NN	O	O
between	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
B	NN	O	O
lymphocyte	NN	O	O
monoclonicity	NN	O	O
or	NN	O	O
lymphoma	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
strong	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
was	NN	O	O
observed	NN	O	O
on	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
labial	NN	O	O
salivary	NN	O	O
glands	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
primary	NN	O	O
SS	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
may	NN	O	O
indicate	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
reactivated	NN	O	O
virus	NN	O	O
hosted	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
lymphocyte	NN	O	B-cell_type
subsets	NN	O	I-cell_type
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
in	NN	O	O
aging	NN	O	O
.	NN	O	O

Plasma	NN	O	O
cortisol	NN	O	O
and	NN	O	O
aldosterone	NN	O	O
levels	NN	O	O
and	NN	O	O
number	NN	O	O
of	NN	O	O
related	NN	O	O
receptors	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
were	NN	O	O
measured	NN	O	O
in	NN	O	O
49	NN	O	O
healthy	NN	O	O
aged	NN	O	O
subjects	NN	O	O
(	NN	O	O
62-97	NN	O	O
yr	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
21	NN	O	O
adult	NN	O	O
controls	NN	O	O
(	NN	O	O
21-50	NN	O	O
yr	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
all	NN	O	O
subjects	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
,	NN	O	O
lymphocyte	NN	O	B-cell_type
subsets	NN	O	I-cell_type
were	NN	O	O
determined	NN	O	O
as	NN	O	O
an	NN	O	O
index	NN	O	O
of	NN	O	O
corticosteroid	NN	O	O
action	NN	O	O
.	NN	O	O

The	NN	O	O
mean	NN	O	O
number	NN	O	O
of	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
aged	NN	O	O
subjects	NN	O	O
than	NN	O	O
in	NN	O	O
controls	NN	O	O
(	NN	O	O
respectively	NN	O	O
,	NN	O	O
198	NN	O	O
+/-	NN	O	O
96	NN	O	O
and	NN	O	O
272	NN	O	O
+/-	NN	O	O
97	NN	O	O
receptors	NN	O	B-protein
/cell	NN	O	O
for	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
,	NN	O	O
and	NN	O	O
1	NN	O	O
,	NN	O	O
794	NN	O	O
+/-	NN	O	O
803	NN	O	O
and	NN	O	O
3	NN	O	O
,	NN	O	O
339	NN	O	O
+/-	NN	O	O
918	NN	O	O
for	NN	O	O
type	NN	O	O
II	NN	O	B-protein
receptors	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Plasma	NN	O	O
aldosterone	NN	O	O
and	NN	O	O
cortisol	NN	O	O
and	NN	O	O
lymphocyte	NN	O	B-cell_type
subsets	NN	O	I-cell_type
were	NN	O	O
not	NN	O	O
different	NN	O	O
in	NN	O	O
the	NN	O	O
two	NN	O	O
groups	NN	O	O
.	NN	O	O

All	NN	O	O
of	NN	O	O
the	NN	O	O
parameters	NN	O	O
were	NN	O	O
also	NN	O	O
tested	NN	O	O
for	NN	O	O
correlation	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
significant	NN	O	O
inverse	NN	O	O
correlation	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
age	NN	O	O
and	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
receptors	NN	O	I-protein
when	NN	O	O
all	NN	O	O
subjects	NN	O	O
were	NN	O	O
plotted	NN	O	O
and	NN	O	O
between	NN	O	O
aged	NN	O	O
and	NN	O	O
CD4	NN	O	B-cell_type
and	NN	O	O
age	NN	O	O
and	NN	O	O
CD4/CD8	NN	O	B-cell_type
in	NN	O	O
the	NN	O	O
aged	NN	O	O
group	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
show	NN	O	O
that	NN	O	O
aged	NN	O	O
subjects	NN	O	O
have	NN	O	O
reductions	NN	O	O
of	NN	O	O
corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
increase	NN	O	O
of	NN	O	O
related	NN	O	O
steroids	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
situation	NN	O	O
probably	NN	O	O
represents	NN	O	O
a	NN	O	O
concomitant	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
aging	NN	O	O
process	NN	O	O
.	NN	O	O

-DOCSTART-	O

T	NN	O	O
cell-specific	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
cytokine	NN	O	I-protein
IL-4	NN	O	I-protein
.	NN	O	O

IL-4	NN	O	B-protein
secreted	NN	O	O
by	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
a	NN	O	O
pleiotropic	NN	O	B-protein
cytokine	NN	O	I-protein
affecting	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
diverse	NN	O	O
cell	NN	O	O
types	NN	O	O
such	NN	O	O
as	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
mast	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
novel	NN	O	B-DNA
T	NN	O	I-DNA
cell-specific	NN	O	I-DNA
negative	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
NRE	NN	O	B-DNA
)	NN	O	O
composed	NN	O	O
of	NN	O	O
two	NN	O	O
protein-binding	NN	O	O
sites	NN	O	O
were	NN	O	O
mapped	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	O
'	NN	O	O
flanking	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
:	NN	O	O
-311CTCCCTTCT-303	NN	O	O
(	NN	O	O
NRE-I	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
-288CTTTTTGCTT-TGC-300	NN	O	O
(	NN	O	O
NRE-II	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

A	NN	O	O
T	NN	O	B-protein
cell-specific	NN	O	I-protein
protein	NN	O	I-protein
Neg-1	NN	O	B-protein
and	NN	O	O
a	NN	O	O
ubiquitous	NN	O	B-protein
protein	NN	O	I-protein
Neg-2	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
NRE-I	NN	O	B-DNA
and	NN	O	O
NRE-II	NN	O	B-DNA
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
a	NN	O	O
positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
was	NN	O	O
found	NN	O	O
45	NN	O	B-DNA
bp	NN	O	I-DNA
downstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
NRE	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
PRE	NN	O	B-DNA
was	NN	O	O
completely	NN	O	O
suppressed	NN	O	O
when	NN	O	O
the	NN	O	O
NRE	NN	O	B-DNA
was	NN	O	O
present	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
IL-4	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
is	NN	O	O
normally	NN	O	O
down-regulated	NN	O	O
by	NN	O	O
an	NN	O	O
NRE	NN	O	B-DNA
via	NN	O	O
repression	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
positive	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
may	NN	O	O
have	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
stringent	NN	O	O
control	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Interleukin	NN	O	O
6-induced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
into	NN	O	O
IgM-secreting	NN	O	B-cell_type
plasma	NN	O	I-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
c-fos	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
protooncogene	NN	O	B-DNA
c-fos	NN	O	I-DNA
in	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
6	NN	O	I-protein
-induced	NN	O	O
B	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
was	NN	O	O
assessed	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
SKW	NN	O	O
6.4	NN	O	O
cells	NN	O	O
with	NN	O	O
IL	NN	O	B-protein
6	NN	O	I-protein
induced	NN	O	O
a	NN	O	O
transient	NN	O	O
and	NN	O	O
early	NN	O	O
stimulation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
sense	NN	O	I-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
appeared	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
and	NN	O	O
returned	NN	O	O
to	NN	O	O
basal	NN	O	O
levels	NN	O	O
after	NN	O	O
2	NN	O	O
h	NN	O	O
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
antisense	NN	O	O
oligonucleotides	NN	O	O
to	NN	O	O
c-fos	NN	O	B-DNA
significantly	NN	O	O
inhibited	NN	O	O
IL	NN	O	B-protein
6	NN	O	I-protein
-induced	NN	O	O
IgM	NN	O	B-protein
production	NN	O	O
by	NN	O	O
SKW	NN	O	B-cell_line
6.4	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0.001	NN	O	O
)	NN	O	O
,	NN	O	O
whereas	NN	O	O
control	NN	O	O
oligonucleotides	NN	O	O
had	NN	O	O
no	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
is	NN	O	O
involved	NN	O	O
in	NN	O	O
IL	NN	O	B-protein
6	NN	O	I-protein
-induced	NN	O	O
differentiation	NN	O	O
of	NN	O	O
SKW	NN	O	B-cell_line
6.4	NN	O	I-cell_line
cells	NN	O	I-cell_line
into	NN	O	O
IgM-secreting	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
regulation	NN	O	O
during	NN	O	O
T-cell	NN	O	B-cell_type
development	NN	O	O
:	NN	O	O
the	NN	O	O
alpha	NN	O	B-DNA
TCR	NN	O	I-DNA
gene	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
molecular	NN	O	O
model	NN	O	O
.	NN	O	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
lymphocyte	NN	O	B-cell_type
differentiation	NN	O	O
is	NN	O	O
a	NN	O	O
complex	NN	O	O
process	NN	O	O
involving	NN	O	O
interactions	NN	O	O
between	NN	O	O
multiple	NN	O	B-DNA
positive	NN	O	I-DNA
and	NN	O	I-DNA
negative	NN	O	I-DNA
transcriptional	NN	O	I-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
article	NN	O	O
,	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
archetypal	NN	O	B-DNA
T-cell-specific	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
alpha	NN	O	B-DNA
TCR	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

Major	NN	O	O
recent	NN	O	O
developments	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
novel	NN	O	O
families	NN	O	O
of	NN	O	O
transcription	NN	O	B-DNA
factors	NN	O	I-DNA
that	NN	O	O
regulate	NN	O	O
multiple	NN	O	O
T-cell	NN	O	B-DNA
genes	NN	O	I-DNA
during	NN	O	O
thymocyte	NN	O	O
ontogeny	NN	O	O
and	NN	O	O
T-cell	NN	O	B-cell_type
activation	NN	O	O
,	NN	O	O
are	NN	O	O
described	NN	O	O
.	NN	O	O

-DOCSTART-	O

Photoaffinity	NN	O	O
labeling	NN	O	O
of	NN	O	O
plasma	NN	O	B-protein
membrane	NN	O	I-protein
receptors	NN	O	I-protein
for	NN	O	O
aldosterone	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

Non-genomic	NN	O	O
effects	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
on	NN	O	O
the	NN	O	O
sodium-proton-antiport	NN	O	B-protein
have	NN	O	O
been	NN	O	O
shown	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
which	NN	O	O
could	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
a	NN	O	O
new	NN	O	O
aldosterone	NN	O	B-protein
membrane	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
paper	NN	O	O
plasma	NN	O	O
membranes	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
were	NN	O	O
covalently	NN	O	O
photolabeled	NN	O	O
with	NN	O	O
a	NN	O	O
[	NN	O	O
125I	NN	O	O
]	NN	O	O
-aldosterone	NN	O	O
derivative	NN	O	O
.	NN	O	O

Sodium	NN	O	O
dodecyl	NN	O	O
sulfate-polyacrylamide	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
showed	NN	O	O
significant	NN	O	O
aldosterone	NN	O	O
binding	NN	O	O
at	NN	O	O
a	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
approximately	NN	O	O
50000	NN	O	O
Dalton	NN	O	O
which	NN	O	O
was	NN	O	O
absent	NN	O	O
with	NN	O	O
1	NN	O	O
microM	NN	O	O
cold	NN	O	O
aldosterone	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
cortisol	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
media	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
sulfhydryl	NN	O	O
agent	NN	O	O
dithiothreitol	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
results	NN	O	O
suggesting	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
disulfide	NN	O	B-protein
bridges	NN	O	I-protein
in	NN	O	O
the	NN	O	O
steroid	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
are	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
the	NN	O	O
membrane	NN	O	B-protein
receptor	NN	O	I-protein
for	NN	O	O
aldosterone	NN	O	O
.	NN	O	O

-DOCSTART-	O

Redox	NN	O	O
status	NN	O	O
of	NN	O	O
cells	NN	O	O
influences	NN	O	O
constitutive	NN	O	O
or	NN	O	O
induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
and	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
and	NN	O	I-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
tested	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
cellular	NN	O	O
activation	NN	O	O
events	NN	O	O
occurring	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
mediated	NN	O	O
through	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
are	NN	O	O
dependent	NN	O	O
upon	NN	O	O
the	NN	O	O
constitutive	NN	O	O
redox	NN	O	O
status	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
phenolic	NN	O	O
,	NN	O	O
lipid-soluble	NN	O	O
,	NN	O	O
chain-breaking	NN	O	O
antioxidants	NN	O	O
(	NN	O	O
butylated	NN	O	O
hydroxyanisole	NN	O	O
(	NN	O	O
BHA	NN	O	O
)	NN	O	O
,	NN	O	O
nordihydroquairetic	NN	O	O
acid	NN	O	O
,	NN	O	O
or	NN	O	O
alpha-tocopherol	NN	O	O
(	NN	O	O
vitamin	NN	O	O
E	NN	O	O
)	NN	O	O
to	NN	O	O
show	NN	O	O
that	NN	O	O
peroxyl	NN	O	O
radical	NN	O	O
scavenging	NN	O	O
in	NN	O	O
unstimulated	NN	O	B-cell_line
and	NN	O	I-cell_line
PMA-	NN	O	I-cell_line
or	NN	O	I-cell_line
TNF-stimulated	NN	O	I-cell_line
cells	NN	O	I-cell_line
blocks	NN	O	O
the	NN	O	O
functions	NN	O	O
depending	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

BHA	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
suppress	NN	O	O
not	NN	O	O
only	NN	O	O
PMA-	NN	O	O
or	NN	O	O
TNF-induced	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
constitutive	NN	O	O
,	NN	O	O
HIV-enhancer	NN	O	O
activity	NN	O	O
concomitant	NN	O	O
to	NN	O	O
an	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
both	NN	O	O
lymphoblastoid	NN	O	B-cell_line
T	NN	O	I-cell_line
(	NN	O	O
J.Jhan	NN	O	B-cell_line
)	NN	O	O
and	NN	O	O
monocytic	NN	O	B-cell_line
(	NN	O	I-cell_line
U937	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
was	NN	O	O
also	NN	O	O
true	NN	O	O
for	NN	O	O
KBF	NN	O	O
(	NN	O	O
p50	NN	O	O
homodimer	NN	O	O
)	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Secretion	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
,	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
another	NN	O	O
NF-kappa	NN	O	B-DNA
B-dependent	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
abolished	NN	O	O
by	NN	O	O
BHA	NN	O	O
in	NN	O	O
PMA-stimulated	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
anti-oxidative	NN	O	O
effect	NN	O	O
of	NN	O	O
BHA	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
thiol	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
glutathione	NN	O	O
,	NN	O	O
content	NN	O	O
in	NN	O	O
stimulated	NN	O	B-cell_line
and	NN	O	I-cell_line
unstimulated	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
TNF	NN	O	B-protein
stimulation	NN	O	O
itself	NN	O	O
barely	NN	O	O
modified	NN	O	O
the	NN	O	O
cellular	NN	O	O
thiol	NN	O	O
level	NN	O	O
.	NN	O	O

Oxidative	NN	O	O
stress	NN	O	O
obtained	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
H2O2	NN	O	O
to	NN	O	O
the	NN	O	O
culture	NN	O	O
medium	NN	O	O
of	NN	O	O
J.Jhan	NN	O	B-cell_line
or	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
could	NN	O	O
not	NN	O	O
by	NN	O	O
itself	NN	O	O
induce	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
TNF	NN	O	B-protein
and	NN	O	O
PMA	NN	O	B-protein
do	NN	O	O
not	NN	O	O
lead	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
through	NN	O	O
induction	NN	O	O
of	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
redox	NN	O	O
status	NN	O	O
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
TNF	NN	O	B-protein
and	NN	O	O
PMA	NN	O	B-protein
can	NN	O	O
exert	NN	O	O
their	NN	O	O
effect	NN	O	O
only	NN	O	O
if	NN	O	O
cells	NN	O	O
are	NN	O	O
in	NN	O	O
an	NN	O	O
appropriate	NN	O	O
redox	NN	O	O
status	NN	O	O
,	NN	O	O
because	NN	O	O
prior	NN	O	O
modification	NN	O	O
toward	NN	O	O
reduction	NN	O	O
with	NN	O	O
BHA	NN	O	O
treatment	NN	O	O
prevents	NN	O	O
this	NN	O	O
activation	NN	O	O
.	NN	O	O

It	NN	O	O
appears	NN	O	O
that	NN	O	O
a	NN	O	O
basal	NN	O	O
redox	NN	O	O
equilibrium	NN	O	O
tending	NN	O	O
toward	NN	O	O
oxidation	NN	O	O
is	NN	O	O
a	NN	O	O
prerequisite	NN	O	O
for	NN	O	O
full	NN	O	O
activation	NN	O	O
of	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
regulating	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B-dependent	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Selection	NN	O	O
of	NN	O	O
optimal	NN	O	O
kappa	NN	O	O
B/Rel	NN	O	O
DNA-binding	NN	O	O
motifs	NN	O	O
:	NN	O	O
interaction	NN	O	O
of	NN	O	O
both	NN	O	O
subunits	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
with	NN	O	O
DNA	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
subunits	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complex	NN	O	I-protein
has	NN	O	O
revealed	NN	O	O
that	NN	O	O
both	NN	O	O
proteins	NN	O	O
can	NN	O	O
interact	NN	O	O
with	NN	O	O
related	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
through	NN	O	O
either	NN	O	O
homo-	NN	O	O
or	NN	O	O
heterodimer	NN	O	O
formation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
proto-oncogene	NN	O	B-DNA
c-rel	NN	O	I-DNA
can	NN	O	O
bind	NN	O	O
to	NN	O	O
similar	NN	O	O
DNA	NN	O	O
motifs	NN	O	O
by	NN	O	O
itself	NN	O	O
or	NN	O	O
as	NN	O	O
a	NN	O	O
heterodimer	NN	O	O
with	NN	O	O
p50	NN	O	B-protein
or	NN	O	O
p65	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
these	NN	O	O
studies	NN	O	O
have	NN	O	O
used	NN	O	O
a	NN	O	O
limited	NN	O	O
number	NN	O	O
of	NN	O	O
known	NN	O	O
kappa	NN	O	O
B	NN	O	O
DNA	NN	O	O
motifs	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
question	NN	O	O
of	NN	O	O
the	NN	O	O
optimal	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
preferred	NN	O	O
by	NN	O	O
each	NN	O	O
homodimer	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
addressed	NN	O	O
.	NN	O	O

Using	NN	O	O
purified	NN	O	O
recombinant	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
and	NN	O	O
c-Rel	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
optimal	NN	O	O
DNA-binding	NN	O	O
motifs	NN	O	O
were	NN	O	O
selected	NN	O	O
from	NN	O	O
a	NN	O	O
pool	NN	O	O
of	NN	O	O
random	NN	O	O
oligonucleotides	NN	O	O
.	NN	O	O

Alignment	NN	O	O
of	NN	O	O
the	NN	O	O
selected	NN	O	O
sequences	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
predict	NN	O	O
a	NN	O	O
consensus	NN	O	O
sequence	NN	O	O
for	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
individual	NN	O	O
homodimeric	NN	O	B-protein
Rel-related	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
and	NN	O	O
DNA-protein	NN	O	O
binding	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
selected	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
revealed	NN	O	O
sequence	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
proteins	NN	O	O
.	NN	O	O

Contrary	NN	O	O
to	NN	O	O
previous	NN	O	O
assumptions	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
p65	NN	O	B-protein
homodimers	NN	O	O
can	NN	O	O
interact	NN	O	O
with	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
not	NN	O	O
recognized	NN	O	O
by	NN	O	O
p50	NN	O	B-protein
homodimers	NN	O	I-protein
.	NN	O	O

Differential	NN	O	O
binding	NN	O	O
affinities	NN	O	O
were	NN	O	O
also	NN	O	O
obtained	NN	O	O
with	NN	O	O
p50-	NN	O	B-DNA
and	NN	O	I-DNA
c-Rel-selected	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
either	NN	O	O
a	NN	O	O
p50-	NN	O	O
or	NN	O	O
p65-	NN	O	O
selected	NN	O	O
kappa	NN	O	O
B	NN	O	O
motif	NN	O	O
,	NN	O	O
which	NN	O	O
displayed	NN	O	O
differential	NN	O	O
binding	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
the	NN	O	O
other	NN	O	O
protein	NN	O	O
,	NN	O	O
little	NN	O	O
to	NN	O	O
no	NN	O	O
binding	NN	O	O
was	NN	O	O
observed	NN	O	O
with	NN	O	O
the	NN	O	O
heterodimeric	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
in	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
selective	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
were	NN	O	O
used	NN	O	O
to	NN	O	O
drive	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
p65	NN	O	B-DNA
-and	NN	O	I-DNA
p50-selected	NN	O	I-DNA
motifs	NN	O	I-DNA
were	NN	O	O
activated	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
p50/65	NN	O	B-protein
(	NN	O	O
a	NN	O	O
chimeric	NN	O	B-protein
protein	NN	O	I-protein
with	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
and	NN	O	O
p65	NN	O	B-protein
activation	NN	O	I-protein
domain	NN	O	I-protein
)	NN	O	O
expression	NN	O	O
vectors	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
and	NN	O	O
neither	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
significant	NN	O	O
response	NN	O	O
to	NN	O	O
stimuli	NN	O	O
that	NN	O	O
induce	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
interaction	NN	O	O
of	NN	O	O
both	NN	O	O
subunits	NN	O	O
of	NN	O	O
the	NN	O	O
heterodimeric	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
complex	NN	O	I-protein
with	NN	O	O
DNA	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
individual	NN	O	O
rel-related	NN	O	B-protein
proteins	NN	O	I-protein
will	NN	O	O
differ	NN	O	O
dramatically	NN	O	O
,	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
specific	NN	O	O
kappa	NN	O	O
B	NN	O	O
motifs	NN	O	O
present	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	B-DNA
subunit	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
underlies	NN	O	O
self-perpetuation	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
transcription	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
the	NN	O	O
sustained	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
observed	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
chronically	NN	O	O
infected	NN	O	O
with	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
regulating	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
the	NN	O	O
p105	NN	O	B-protein
precursor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	I-protein
subunit	NN	O	I-protein
was	NN	O	O
enhanced	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Deletions	NN	O	O
in	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
indicated	NN	O	O
that	NN	O	O
this	NN	O	O
upregulation	NN	O	O
was	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
NF-kappa	NN	O	O
B-	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
AP-1-binding	NN	O	B-DNA
motif	NN	O	I-DNA
,	NN	O	O
by	NN	O	O
bona	NN	O	O
fide	NN	O	O
p50/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
cytosolic	NN	O	O
extracts	NN	O	O
indicated	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
levels	NN	O	O
were	NN	O	O
increased	NN	O	O
in	NN	O	O
HIV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
transient	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
,	NN	O	O
the	NN	O	O
permanent	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
induced	NN	O	O
by	NN	O	O
HIV	NN	O	O
infection	NN	O	O
was	NN	O	O
not	NN	O	O
dependent	NN	O	O
on	NN	O	O
PKC	NN	O	B-protein
isoenzymes	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
beta	NN	O	I-protein
as	NN	O	O
shown	NN	O	O
by	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
a	NN	O	O
specific	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
GF	NN	O	O
109203X	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
indicate	NN	O	O
that	NN	O	O
during	NN	O	O
chronic	NN	O	O
HIV	NN	O	O
infection	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
continuous	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
p50/p65	NN	O	B-protein
)	NN	O	O
translocation	NN	O	O
results	NN	O	O
in	NN	O	O
p105	NN	O	O
promoter	NN	O	O
upregulation	NN	O	O
with	NN	O	O
subsequent	NN	O	O
cytosolic	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
accumulation	NN	O	O
,	NN	O	O
ready	NN	O	O
for	NN	O	O
further	NN	O	O
translocation	NN	O	O
.	NN	O	O

This	NN	O	O
HIV-mediated	NN	O	O
mechanism	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
self-perpetuating	NN	O	O
loop	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
production	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
acts	NN	O	O
on	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
by	NN	O	O
enhancing	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
to	NN	O	O
its	NN	O	O
consensus	NN	O	O
sequence	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	O
'	NN	O	O
regulatory	NN	O	O
enhancer	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
chain	NN	O	I-DNA
(	NN	O	O
p55	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
activates	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U	NN	O	I-cell_line
937	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
human	NN	O	I-cell_type
T-cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
effect	NN	O	O
is	NN	O	O
detectable	NN	O	O
within	NN	O	O
15	NN	O	O
min	NN	O	O
and	NN	O	O
peaks	NN	O	O
1	NN	O	O
h	NN	O	O
after	NN	O	O
exposure	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
.	NN	O	O

Enhanced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
followed	NN	O	O
by	NN	O	O
functional	NN	O	O
activation	NN	O	O
in	NN	O	O
that	NN	O	O
inducibility	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
enhanced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
and	NN	O	O
that	NN	O	O
a	NN	O	O
heterologous	NN	O	O
promoter	NN	O	O
containing	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
consensus	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
-291	NN	O	B-DNA
to	NN	O	I-DNA
-245	NN	O	I-DNA
)	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
activated	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
increasing	NN	O	O
transcript	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	O
factor	NN	O	O
,	NN	O	O
whereas	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
gene	NN	O	I-DNA
remained	NN	O	O
unchanged	NN	O	O
.	NN	O	O

-DOCSTART-	O

Studies	NN	O	O
on	NN	O	O
the	NN	O	O
biological	NN	O	O
activity	NN	O	O
of	NN	O	O
triiodothyronine	NN	O	O
sulfate	NN	O	O
.	NN	O	O

Hepatic	NN	O	B-cell_type
microsomes	NN	O	I-cell_type
and	NN	O	O
isolated	NN	O	B-cell_type
hepatocytes	NN	O	I-cell_type
in	NN	O	O
short	NN	O	O
term	NN	O	O
culture	NN	O	O
desulfate	NN	O	O
T3	NN	O	O
sulfate	NN	O	O
(	NN	O	O
T3SO4	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
wished	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
T3SO4	NN	O	O
could	NN	O	O
mimic	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
thyroid	NN	O	O
hormone	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

T3SO4	NN	O	O
had	NN	O	O
no	NN	O	O
thyromimetic	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
Ca	NN	O	B-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-ATPase	NN	O	I-protein
in	NN	O	O
human	NN	O	O
erythrocyte	NN	O	O
membranes	NN	O	O
at	NN	O	O
doses	NN	O	O
up	NN	O	O
to	NN	O	O
10	NN	O	O
,	NN	O	O
000	NN	O	O
times	NN	O	O
the	NN	O	O
maximally	NN	O	O
effective	NN	O	O
dose	NN	O	O
of	NN	O	O
T3	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-10	NN	O	O
)	NN	O	O
mol/L	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
GH4C1	NN	O	B-cell_line
pituitary	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
T3SO4	NN	O	O
failed	NN	O	O
to	NN	O	O
displace	NN	O	O
[	NN	O	O
125I	NN	O	O
]	NN	O	O
T3	NN	O	O
from	NN	O	O
nuclear	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
intact	NN	O	B-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
soluble	NN	O	O
preparations	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
T3SO4	NN	O	O
was	NN	O	O
not	NN	O	O
directly	NN	O	O
thyromimetic	NN	O	O
in	NN	O	O
either	NN	O	O
an	NN	O	O
isolated	NN	O	O
human	NN	O	O
membrane	NN	O	O
system	NN	O	O
or	NN	O	O
a	NN	O	O
pituitary	NN	O	O
cell	NN	O	O
system	NN	O	O
in	NN	O	O
which	NN	O	O
nuclear	NN	O	O
receptor	NN	O	O
occupancy	NN	O	O
correlates	NN	O	O
with	NN	O	O
GH	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Thyroid	NN	O	O
hormones	NN	O	O
inhibit	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
glycosaminoglycan	NN	O	O
synthesis	NN	O	O
by	NN	O	O
cultured	NN	O	B-cell_line
human	NN	O	I-cell_line
dermal	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
T3SO4	NN	O	O
displayed	NN	O	O
about	NN	O	O
0.5	NN	O	O
%	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
T3	NN	O	O
at	NN	O	O
72	NN	O	O
h	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
contained	NN	O	O
roughly	NN	O	O
the	NN	O	O
same	NN	O	O
level	NN	O	O
of	NN	O	O
microsomal	NN	O	O
p-nitrophenyl	NN	O	B-protein
sulfatase	NN	O	I-protein
activity	NN	O	O
as	NN	O	O
that	NN	O	O
previously	NN	O	O
observed	NN	O	O
in	NN	O	O
hepatic	NN	O	B-cell_type
microsomes	NN	O	I-cell_type
.	NN	O	O

Propylthiouracil	NN	O	O
(	NN	O	O
50	NN	O	O
mumol/L	NN	O	O
)	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
T3SO4	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
deiodination	NN	O	O
was	NN	O	O
not	NN	O	O
important	NN	O	O
for	NN	O	O
this	NN	O	O
activity	NN	O	O
of	NN	O	O
T3SO4	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
it	NN	O	O
appears	NN	O	O
T3SO4	NN	O	O
has	NN	O	O
no	NN	O	O
intrinsic	NN	O	O
biological	NN	O	O
activity	NN	O	O
,	NN	O	O
but	NN	O	O
,	NN	O	O
under	NN	O	O
certain	NN	O	O
circumstances	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
reactivated	NN	O	O
by	NN	O	O
desulfation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
through	NN	O	O
the	NN	O	O
activation-inducer	NN	O	B-protein
molecule/CD69	NN	O	I-protein
enhances	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
has	NN	O	O
been	NN	O	O
ascribed	NN	O	O
to	NN	O	O
the	NN	O	O
early	NN	O	O
events	NN	O	O
leading	NN	O	O
to	NN	O	O
T	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
stimulated	NN	O	O
through	NN	O	O
the	NN	O	O
activation	NN	O	O
inducer	NN	O	O
molecule	NN	O	O
(	NN	O	O
AIM	NN	O	O
)	NN	O	O
/	NN	O	O
CD69	NN	O	B-protein
activation	NN	O	O
pathway	NN	O	O
.	NN	O	O

Phorbol	NN	O	O
esters	NN	O	O
are	NN	O	O
required	NN	O	O
to	NN	O	O
induce	NN	O	O
AIM/CD69	NN	O	B-protein
cell-surface	NN	O	O
expression	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
for	NN	O	O
triggering	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
anti-AIM	NN	O	B-protein
mAb	NN	O	I-protein
.	NN	O	O

Mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
addition	NN	O	O
of	NN	O	O
anti-AIM	NN	O	B-protein
mAb	NN	O	I-protein
to	NN	O	O
PMA-treated	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
markedly	NN	O	O
enhanced	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
to	NN	O	O
its	NN	O	O
cognate	NN	O	O
sequence	NN	O	O
,	NN	O	O
the	NN	O	O
phorbol	NN	O	B-DNA
ester	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
anti-AIM	NN	O	B-protein
mAb	NN	O	I-protein
did	NN	O	O
not	NN	O	O
induce	NN	O	O
any	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
whose	NN	O	O
activity	NN	O	O
is	NN	O	O
also	NN	O	O
regulated	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
.	NN	O	O

The	NN	O	O
increase	NN	O	O
in	NN	O	O
AP-1	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
the	NN	O	O
marked	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
c-fos	NN	O	B-protein
but	NN	O	O
not	NN	O	O
that	NN	O	O
of	NN	O	O
c-jun	NN	O	B-protein
.	NN	O	O

Blockade	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
complexes	NN	O	I-protein
with	NN	O	O
an	NN	O	O
anti-Fos	NN	O	B-protein
mAb	NN	O	I-protein
demonstrated	NN	O	O
a	NN	O	O
direct	NN	O	O
participation	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
in	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
complexes	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
anti-AIM	NN	O	B-protein
mAb	NN	O	I-protein
.	NN	O	O

Most	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
could	NN	O	O
be	NN	O	O
eliminated	NN	O	O
when	NN	O	O
the	NN	O	O
anti-AIM	NN	O	B-protein
mAb	NN	O	I-protein
was	NN	O	O
added	NN	O	O
to	NN	O	O
the	NN	O	O
culture	NN	O	O
medium	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cycloheximide	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
provide	NN	O	O
the	NN	O	O
evidence	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
through	NN	O	O
the	NN	O	O
AIM	NN	O	O
activation	NN	O	O
pathway	NN	O	O
regulate	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
this	NN	O	O
pathway	NN	O	O
appears	NN	O	O
as	NN	O	O
a	NN	O	O
crucial	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
early	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
events	NN	O	O
.	NN	O	O

-DOCSTART-	O

Gangliosides	NN	O	O
suppress	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
production	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Both	NN	O	O
normal	NN	O	B-cell_type
and	NN	O	I-cell_type
malignant	NN	O	I-cell_type
cells	NN	O	I-cell_type
contain	NN	O	O
gangliosides	NN	O	O
as	NN	O	O
important	NN	O	O
cell	NN	O	O
membrane	NN	O	O
constituents	NN	O	O
that	NN	O	O
,	NN	O	O
after	NN	O	O
being	NN	O	O
shed	NN	O	O
,	NN	O	O
may	NN	O	O
influence	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
impact	NN	O	O
of	NN	O	O
gangliosides	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
in	NN	O	O
blood	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Mono	NN	O	I-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
.	NN	O	O

Although	NN	O	O
under	NN	O	O
standard	NN	O	O
culture	NN	O	O
conditions	NN	O	O
,	NN	O	O
bovine	NN	O	O
brain	NN	O	O
gangliosides	NN	O	O
(	NN	O	O
100	NN	O	O
micrograms/ml	NN	O	O
)	NN	O	O
suppressed	NN	O	O
LPS-stimulated	NN	O	O
TNF	NN	O	B-protein
production	NN	O	O
5-fold	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
and	NN	O	O
10-fold	NN	O	O
in	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suppression	NN	O	O
was	NN	O	O
more	NN	O	O
efficient	NN	O	O
under	NN	O	O
serum-free	NN	O	O
conditions	NN	O	O
.	NN	O	O

Looking	NN	O	O
at	NN	O	O
highly	NN	O	O
purified	NN	O	O
gangliosides	NN	O	O
,	NN	O	O
GD3	NN	O	O
,	NN	O	O
GD1a	NN	O	O
,	NN	O	O
GM3	NN	O	O
,	NN	O	O
GM2	NN	O	O
,	NN	O	O
and	NN	O	O
GM1	NN	O	O
were	NN	O	O
all	NN	O	O
effective	NN	O	O
in	NN	O	O
reducing	NN	O	O
TNF	NN	O	B-protein
production	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
in	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
by	NN	O	O
factor	NN	O	O
10	NN	O	O
to	NN	O	O
50	NN	O	O
.	NN	O	O

The	NN	O	O
suppressive	NN	O	O
activity	NN	O	O
was	NN	O	O
lost	NN	O	O
in	NN	O	O
molecules	NN	O	O
,	NN	O	O
lacking	NN	O	O
the	NN	O	O
sugar	NN	O	O
moiety	NN	O	O
or	NN	O	O
the	NN	O	O
lipid	NN	O	O
moiety	NN	O	O
.	NN	O	O

Gangliosides	NN	O	O
appear	NN	O	O
to	NN	O	O
act	NN	O	O
at	NN	O	O
an	NN	O	O
early	NN	O	O
step	NN	O	O
of	NN	O	O
activation	NN	O	O
in	NN	O	O
that	NN	O	O
TNF	NN	O	B-RNA
transcripts	NN	O	I-RNA
were	NN	O	O
reduced	NN	O	O
and	NN	O	O
the	NN	O	O
mobilization	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
was	NN	O	O
blocked	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
in	NN	O	O
time	NN	O	O
kinetics	NN	O	O
,	NN	O	O
gangliosides	NN	O	O
were	NN	O	O
effective	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
30	NN	O	O
min	NN	O	O
after	NN	O	O
addition	NN	O	O
of	NN	O	O
LPS	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
thereafter	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CD14	NN	O	B-protein
Ag	NN	O	I-protein
,	NN	O	O
a	NN	O	O
receptor	NN	O	B-protein
molecule	NN	O	I-protein
for	NN	O	O
LPS-LPS	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
complexes	NN	O	I-protein
,	NN	O	O
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
gangliosides	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
when	NN	O	O
using	NN	O	O
Staphylococcus	NN	O	O
aureus	NN	O	O
or	NN	O	O
platelet	NN	O	B-protein
activating	NN	O	I-protein
factor	NN	O	I-protein
as	NN	O	O
a	NN	O	O
stimulus	NN	O	O
,	NN	O	O
gangliosides	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
suppress	NN	O	O
TNF	NN	O	B-protein
production	NN	O	O
in	NN	O	O
Mono	NN	O	B-cell_line
Mac	NN	O	I-cell_line
6	NN	O	I-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
factor	NN	O	O
5	NN	O	O
to	NN	O	O
10	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
other	NN	O	O
hand	NN	O	O
,	NN	O	O
phorbol	NN	O	O
ester-induced	NN	O	O
production	NN	O	O
of	NN	O	O
O2-	NN	O	O
was	NN	O	O
similar	NN	O	O
in	NN	O	O
cells	NN	O	O
treated	NN	O	O
with	NN	O	O
and	NN	O	O
without	NN	O	O
gangliosides	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
TNF	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
induced	NN	O	O
by	NN	O	O
different	NN	O	O
types	NN	O	O
of	NN	O	O
stimuli	NN	O	O
can	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
gangliosides	NN	O	O
at	NN	O	O
an	NN	O	O
early	NN	O	O
step	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
mitogen-inducible	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
related	NN	O	O
to	NN	O	O
p50/p105-NF-kappa	NN	O	B-protein
B	NN	O	I-protein
participates	NN	O	O
in	NN	O	O
transactivation	NN	O	O
through	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
Rel-related	NN	O	B-protein
,	NN	O	I-protein
mitogen-inducible	NN	O	I-protein
,	NN	O	I-protein
kappa	NN	O	I-protein
B-binding	NN	O	I-protein
protein	NN	O	I-protein
has	NN	O	O
been	NN	O	O
cloned	NN	O	O
as	NN	O	O
an	NN	O	O
immediate-early	NN	O	B-DNA
activation	NN	O	I-DNA
gene	NN	O	I-DNA
of	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
cDNA	NN	O	B-DNA
has	NN	O	O
an	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
of	NN	O	O
900	NN	O	B-DNA
amino	NN	O	I-DNA
acids	NN	O	I-DNA
capable	NN	O	O
of	NN	O	O
encoding	NN	O	O
a	NN	O	O
97-kDa	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

This	NN	O	O
protein	NN	O	O
is	NN	O	O
most	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
105-kDa	NN	O	O
precursor	NN	O	O
polypeptide	NN	O	O
of	NN	O	O
p50-NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Like	NN	O	O
the	NN	O	O
105-kDa	NN	O	O
precursor	NN	O	O
,	NN	O	O
it	NN	O	O
contains	NN	O	O
an	NN	O	O
amino-terminal	NN	O	B-protein
Rel-related	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
about	NN	O	O
300	NN	O	O
amino	NN	O	O
acids	NN	O	O
and	NN	O	O
a	NN	O	O
carboxy-terminal	NN	O	B-protein
domain	NN	O	I-protein
containing	NN	O	O
six	NN	O	O
full	NN	O	B-protein
cell	NN	O	I-protein
cycle	NN	O	I-protein
or	NN	O	O
ankyrin	NN	O	B-protein
repeats	NN	O	I-protein
.	NN	O	O

In	NN	O	O
vitro-translated	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
truncated	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
Rel	NN	O	B-DNA
domain	NN	O	I-DNA
and	NN	O	O
excluding	NN	O	O
the	NN	O	O
repeats	NN	O	O
,	NN	O	O
bind	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
refer	NN	O	O
to	NN	O	O
the	NN	O	O
kappa	NN	O	B-protein
B-binding	NN	O	I-protein
,	NN	O	I-protein
truncated	NN	O	I-protein
protein	NN	O	I-protein
as	NN	O	O
p50B	NN	O	B-protein
by	NN	O	O
analogy	NN	O	O
with	NN	O	O
p50-NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
to	NN	O	O
the	NN	O	O
full-length	NN	O	O
protein	NN	O	O
as	NN	O	O
p97	NN	O	B-protein
.	NN	O	O

p50B	NN	O	B-protein
is	NN	O	O
able	NN	O	O
to	NN	O	O
form	NN	O	O
heteromeric	NN	O	B-protein
kappa	NN	O	I-protein
B-binding	NN	O	I-protein
complexes	NN	O	I-protein
with	NN	O	O
RelB	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
with	NN	O	O
p65	NN	O	B-protein
and	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
the	NN	O	O
two	NN	O	O
subunits	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Transient-transfection	NN	O	O
experiments	NN	O	O
in	NN	O	O
embryonal	NN	O	B-cell_line
carcinoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
demonstrate	NN	O	O
a	NN	O	O
functional	NN	O	O
cooperation	NN	O	O
between	NN	O	O
p50B	NN	O	B-protein
and	NN	O	O
RelB	NN	O	B-protein
or	NN	O	O
p65	NN	O	B-protein
in	NN	O	O
transactivation	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
plasmid	NN	O	I-DNA
dependent	NN	O	O
on	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
data	NN	O	O
imply	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
complex	NN	O	O
family	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B-like	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Surrogate	NN	O	O
thyroglobulin	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
in	NN	O	O
Hashimoto	NN	O	O
's	NN	O	O
thyroiditis	NN	O	O
.	NN	O	O

Immunoglobulin	NN	O	B-protein
molecules	NN	O	I-protein
on	NN	O	O
the	NN	O	O
surface	NN	O	O
of	NN	O	O
a	NN	O	O
B	NN	O	B-cell_type
lymphocyte	NN	O	I-cell_type
are	NN	O	O
the	NN	O	O
endogenous	NN	O	O
``	NN	O	O
receptors	NN	O	O
''	NN	O	O
to	NN	O	O
which	NN	O	O
specific	NN	O	O
antigens	NN	O	O
bind	NN	O	O
.	NN	O	O

Studies	NN	O	O
in	NN	O	O
mice	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
,	NN	O	O
conjugated	NN	O	O
with	NN	O	O
palmitate	NN	O	O
to	NN	O	O
provide	NN	O	O
a	NN	O	O
lipid	NN	O	O
tail	NN	O	O
,	NN	O	O
can	NN	O	O
be	NN	O	O
inserted	NN	O	O
into	NN	O	O
the	NN	O	O
cell	NN	O	O
membrane	NN	O	O
to	NN	O	O
provide	NN	O	O
a	NN	O	O
``	NN	O	O
surrogate	NN	O	O
''	NN	O	O
antigen	NN	O	O
receptor	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
whether	NN	O	O
a	NN	O	O
palmitate	NN	O	B-protein
conjugate	NN	O	I-protein
of	NN	O	O
a	NN	O	O
human	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
specific	NN	O	O
for	NN	O	O
thyroglobulin	NN	O	B-protein
(	NN	O	O
TG	NN	O	B-protein
)	NN	O	O
could	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
surrogate	NN	O	B-protein
TG	NN	O	I-protein
receptor	NN	O	I-protein
on	NN	O	O
blood	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
separated	NN	O	O
into	NN	O	O
fractions	NN	O	O
enriched	NN	O	O
for	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
or	NN	O	O
depleted	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
non-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
.	NN	O	O

Using	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
,	NN	O	O
we	NN	O	O
detected	NN	O	O
surrogate	NN	O	B-protein
TG	NN	O	I-protein
receptors	NN	O	I-protein
on	NN	O	O
non-T	NN	O	B-cell_type
(	NN	O	I-cell_type
but	NN	O	I-cell_type
not	NN	O	I-cell_type
on	NN	O	I-cell_type
T	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
11	NN	O	O
of	NN	O	O
11	NN	O	O
individuals	NN	O	O
studied	NN	O	O
(	NN	O	O
5	NN	O	O
Hashimoto	NN	O	O
patients	NN	O	O
and	NN	O	O
6	NN	O	O
control	NN	O	O
donors	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
endogenous	NN	O	B-protein
TG	NN	O	I-protein
receptors	NN	O	I-protein
could	NN	O	O
only	NN	O	O
be	NN	O	O
detected	NN	O	O
on	NN	O	O
non-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
1	NN	O	O
of	NN	O	O
3	NN	O	O
Hashimoto	NN	O	O
patients	NN	O	O
and	NN	O	O
from	NN	O	O
0	NN	O	O
of	NN	O	O
4	NN	O	O
control	NN	O	O
donors	NN	O	O
.	NN	O	O

Because	NN	O	O
of	NN	O	O
the	NN	O	O
efficient	NN	O	O
binding	NN	O	O
of	NN	O	O
TG	NN	O	B-protein
by	NN	O	O
surrogate	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
non-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
assessed	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
such	NN	O	O
cells	NN	O	O
to	NN	O	O
present	NN	O	O
TG	NN	O	B-protein
to	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Proliferation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
TG	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
only	NN	O	O
1	NN	O	O
of	NN	O	O
5	NN	O	O
Hashimoto	NN	O	O
patients	NN	O	O
.	NN	O	O

This	NN	O	O
low	NN	O	O
frequency	NN	O	O
of	NN	O	O
response	NN	O	O
was	NN	O	O
no	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
previously	NN	O	O
detected	NN	O	O
using	NN	O	O
cultures	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
autologous	NN	O	B-cell_type
dendritic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
successful	NN	O	O
generation	NN	O	O
of	NN	O	O
surrogate	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
non-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
more	NN	O	O
efficient	NN	O	O
TG	NN	O	B-protein
presentation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
significance	NN	O	O
of	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
is	NN	O	O
that	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
not	NN	O	O
the	NN	O	O
antigen-presenting	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
are	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
limiting	NN	O	O
element	NN	O	O
in	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferative	NN	O	O
response	NN	O	O
to	NN	O	O
TG	NN	O	B-protein
and	NN	O	O
other	NN	O	O
thyroid	NN	O	B-protein
autoantigens	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Targeted	NN	O	O
degradation	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
v-Fos	NN	O	B-protein
,	NN	O	O
by	NN	O	O
a	NN	O	O
phosphorylation-dependent	NN	O	B-protein
signal	NN	O	I-protein
on	NN	O	O
c-Jun	NN	O	B-protein
.	NN	O	O

The	NN	O	O
proto-oncogene	NN	O	B-protein
products	NN	O	I-protein
c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
heterodimerize	NN	O	O
through	NN	O	O
their	NN	O	O
leucine	NN	O	B-protein
zippers	NN	O	I-protein
to	NN	O	O
form	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
heterodimer	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
signal-dependent	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
dephosphorylation	NN	O	O
events	NN	O	O
.	NN	O	O

The	NN	O	O
stability	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
also	NN	O	O
be	NN	O	O
controlled	NN	O	O
by	NN	O	O
intracellular	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

In	NN	O	O
transient	NN	O	O
expression	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
degradation	NN	O	O
experiments	NN	O	O
,	NN	O	O
the	NN	O	O
stability	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
was	NN	O	O
decreased	NN	O	O
when	NN	O	O
the	NN	O	O
protein	NN	O	O
was	NN	O	O
dimerized	NN	O	O
with	NN	O	O
phosphorylated	NN	O	B-protein
c-Jun	NN	O	I-protein
.	NN	O	O

c-Jun	NN	O	B-protein
protein	NN	O	O
isolated	NN	O	O
from	NN	O	O
phorbol	NN	O	B-cell_line
ester-induced	NN	O	I-cell_line
cells	NN	O	I-cell_line
did	NN	O	O
not	NN	O	O
target	NN	O	O
c-Fos	NN	O	B-protein
for	NN	O	O
degradation	NN	O	O
,	NN	O	O
which	NN	O	O
suggests	NN	O	O
that	NN	O	O
c-Fos	NN	O	B-protein
is	NN	O	O
transiently	NN	O	O
stabilized	NN	O	O
after	NN	O	O
stimulation	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

v-Fos	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
the	NN	O	O
retroviral	NN	O	O
counterpart	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
was	NN	O	O
not	NN	O	O
susceptible	NN	O	O
to	NN	O	O
degradation	NN	O	O
targeted	NN	O	O
by	NN	O	O
c-Jun	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
tumor	NN	O	I-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
in	NN	O	O
macrophage	NN	O	B-cell_line
,	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
1311-base	NN	O	B-DNA
pair	NN	O	I-DNA
human	NN	O	I-DNA
tumor	NN	O	I-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
TNF	NN	O	I-DNA
)	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
was	NN	O	O
fused	NN	O	O
to	NN	O	O
the	NN	O	O
luciferase	NN	O	B-DNA
(	NN	O	I-DNA
Luc	NN	O	I-DNA
)	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
studied	NN	O	O
in	NN	O	O
a	NN	O	O
transient	NN	O	O
transfection	NN	O	O
system	NN	O	O
in	NN	O	O
three	NN	O	O
TNF	NN	O	B-cell_line
producing	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
U937	NN	O	B-cell_line
macrophage	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
MLA	NN	O	B-cell_line
144	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
the	NN	O	O
729-6	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
full	NN	O	O
length	NN	O	O
promoter	NN	O	O
construct	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
13-myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
in	NN	O	O
each	NN	O	O
of	NN	O	O
these	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
5'-truncations	NN	O	B-DNA
showed	NN	O	O
several	NN	O	O
peaks	NN	O	O
of	NN	O	O
basal	NN	O	O
and	NN	O	O
PMA	NN	O	O
induced	NN	O	O
activity	NN	O	O
suggesting	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
several	NN	O	O
positive	NN	O	B-DNA
and	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
elements	NN	O	O
.	NN	O	O

A	NN	O	O
PMA	NN	O	B-DNA
responsive	NN	O	I-DNA
element	NN	O	I-DNA
was	NN	O	O
localized	NN	O	O
to	NN	O	O
a	NN	O	O
region	NN	O	O
between	NN	O	O
-95	NN	O	B-DNA
and	NN	O	I-DNA
-36	NN	O	I-DNA
bp	NN	O	I-DNA
relative	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Within	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
single	NN	O	O
AP-2-	NN	O	B-DNA
and	NN	O	I-DNA
AP-1-	NN	O	I-DNA
like	NN	O	I-DNA
consensus	NN	O	I-DNA
sequences	NN	O	I-DNA
were	NN	O	O
noted	NN	O	O
.	NN	O	O

These	NN	O	O
AP-2	NN	O	B-DNA
and	NN	O	O
AP-1	NN	O	B-DNA
sites	NN	O	I-DNA
were	NN	O	O
each	NN	O	O
modified	NN	O	O
with	NN	O	O
a	NN	O	O
double	NN	O	O
point	NN	O	O
mutation	NN	O	O
.	NN	O	O

A	NN	O	O
modest	NN	O	O
(	NN	O	O
20-50	NN	O	O
%	NN	O	O
)	NN	O	O
reduction	NN	O	O
in	NN	O	O
TNF	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
was	NN	O	O
observed	NN	O	O
with	NN	O	O
the	NN	O	O
AP-2	NN	O	B-DNA
site	NN	O	I-DNA
mutation	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
markedly	NN	O	O
diminished	NN	O	O
both	NN	O	O
the	NN	O	O
basal	NN	O	O
and	NN	O	O
PMA-activated	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

Also	NN	O	O
co-transfections	NN	O	O
of	NN	O	O
the	NN	O	O
wild-type	NN	O	B-DNA
promoter	NN	O	I-DNA
construct	NN	O	I-DNA
with	NN	O	O
an	NN	O	O
AP-1/c-jun	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
augmented	NN	O	O
basal	NN	O	O
and	NN	O	O
PMA-induced	NN	O	O
promoter	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stable	NN	O	O
expression	NN	O	O
of	NN	O	O
transdominant	NN	O	B-protein
Rev	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
inhibits	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
replication	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
Rev	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
viral	NN	O	O
structural	NN	O	O
protein	NN	O	O
expression	NN	O	O
(	NN	O	O
Gag	NN	O	B-protein
,	NN	O	O
Pol	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Env	NN	O	B-protein
)	NN	O	O
and	NN	O	O
,	NN	O	O
hence	NN	O	O
,	NN	O	O
for	NN	O	O
viral	NN	O	O
replication	NN	O	O
.	NN	O	O

In	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
,	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
Rev	NN	O	B-protein
have	NN	O	O
been	NN	O	O
identified	NN	O	O
that	NN	O	O
inhibit	NN	O	O
wild-type	NN	O	O
Rev	NN	O	B-protein
activity	NN	O	O
and	NN	O	O
therefore	NN	O	O
suppress	NN	O	O
viral	NN	O	O
replication	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
such	NN	O	O
transdominant	NN	O	B-protein
Rev	NN	O	I-protein
proteins	NN	O	I-protein
could	NN	O	O
provide	NN	O	O
long-term	NN	O	O
protection	NN	O	O
against	NN	O	O
HIV	NN	O	O
infection	NN	O	O
without	NN	O	O
affecting	NN	O	O
T	NN	O	O
cell	NN	O	O
function	NN	O	O
,	NN	O	O
T	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
stably	NN	O	O
transduced	NN	O	O
with	NN	O	O
a	NN	O	O
retroviral	NN	O	B-DNA
vector	NN	O	I-DNA
encoding	NN	O	O
a	NN	O	O
transdominant	NN	O	O
mutant	NN	O	O
of	NN	O	O
the	NN	O	O
Rev	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
M10	NN	O	B-protein
.	NN	O	O

While	NN	O	O
all	NN	O	O
the	NN	O	O
M10-expressing	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
remained	NN	O	O
infectable	NN	O	O
by	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
these	NN	O	O
same	NN	O	O
cells	NN	O	O
failed	NN	O	O
to	NN	O	O
support	NN	O	O
a	NN	O	O
productive	NN	O	O
replication	NN	O	O
cycle	NN	O	O
when	NN	O	O
infected	NN	O	O
with	NN	O	O
a	NN	O	O
cloned	NN	O	O
isolate	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
two	NN	O	O
out	NN	O	O
of	NN	O	O
three	NN	O	O
M10-expressing	NN	O	B-cell_line
CEM	NN	O	I-cell_line
clones	NN	O	I-cell_line
were	NN	O	O
also	NN	O	O
resistant	NN	O	O
to	NN	O	O
highly	NN	O	O
productive	NN	O	O
infection	NN	O	O
by	NN	O	O
a	NN	O	O
heterogeneous	NN	O	O
HIV-1	NN	O	O
pool	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
M10	NN	O	B-protein
did	NN	O	O
not	NN	O	O
affect	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
or	NN	O	O
Tat	NN	O	B-protein
.	NN	O	O

Importantly	NN	O	O
,	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
Rev	NN	O	B-protein
M10	NN	O	I-protein
did	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
secretion	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogen	NN	O	O
stimulation	NN	O	O
of	NN	O	O
EL-4	NN	O	B-cell_line
and	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
inhibition	NN	O	O
of	NN	O	O
HIV	NN	O	O
infection	NN	O	O
in	NN	O	O
cells	NN	O	O
stably	NN	O	O
expressing	NN	O	O
a	NN	O	O
transdominant	NN	O	B-protein
Rev	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
any	NN	O	O
deleterious	NN	O	O
effect	NN	O	O
on	NN	O	O
T	NN	O	O
cell	NN	O	O
function	NN	O	O
,	NN	O	O
suggests	NN	O	O
that	NN	O	O
such	NN	O	O
a	NN	O	O
strategy	NN	O	O
could	NN	O	O
provide	NN	O	O
a	NN	O	O
therapeutic	NN	O	O
effect	NN	O	O
in	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
of	NN	O	O
acquired	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Simple	NN	O	O
derivation	NN	O	O
of	NN	O	O
TFIID-dependent	NN	O	O
RNA	NN	O	B-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
transcription	NN	O	O
systems	NN	O	O
from	NN	O	O
Schizosaccharomyces	NN	O	O
pombe	NN	O	O
and	NN	O	O
other	NN	O	O
organisms	NN	O	O
,	NN	O	O
and	NN	O	O
factors	NN	O	O
required	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Resolution	NN	O	O
of	NN	O	O
whole	NN	O	O
cell	NN	O	O
extract	NN	O	O
through	NN	O	O
two	NN	O	O
chromatographic	NN	O	O
steps	NN	O	O
yields	NN	O	O
a	NN	O	O
single	NN	O	O
protein	NN	O	O
fraction	NN	O	O
requiring	NN	O	O
only	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
TFIID	NN	O	B-protein
for	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
transcription	NN	O	O
at	NN	O	O
RNA	NN	O	B-DNA
polymerase	NN	O	I-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
approach	NN	O	O
allows	NN	O	O
the	NN	O	O
convenient	NN	O	O
generation	NN	O	O
of	NN	O	O
RNA	NN	O	B-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
transcription	NN	O	O
systems	NN	O	O
from	NN	O	O
Saccharomyces	NN	O	O
cerevisiae	NN	O	O
,	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
Schizosaccharomyces	NN	O	O
pombe	NN	O	O
.	NN	O	O

TFIIDs	NN	O	B-protein
from	NN	O	O
all	NN	O	O
three	NN	O	O
organisms	NN	O	O
are	NN	O	O
interchangeable	NN	O	O
among	NN	O	O
all	NN	O	O
three	NN	O	O
systems	NN	O	O
.	NN	O	O

The	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
and	NN	O	O
Sch.	NN	O	O
pombe	NN	O	O
systems	NN	O	O
support	NN	O	O
effects	NN	O	O
of	NN	O	O
acidic	NN	O	B-protein
activator	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
provided	NN	O	O
a	NN	O	O
further	NN	O	O
protein	NN	O	O
fraction	NN	O	O
from	NN	O	O
S.	NN	O	O
cerevisiae	NN	O	O
is	NN	O	O
supplied	NN	O	O
.	NN	O	O

This	NN	O	O
further	NN	O	O
fraction	NN	O	O
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
mediator	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
described	NN	O	O
previously	NN	O	O
and	NN	O	O
represents	NN	O	O
a	NN	O	O
second	NN	O	O
component	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
general	NN	O	B-protein
initiation	NN	O	I-protein
factors	NN	O	I-protein
that	NN	O	O
may	NN	O	O
facilitate	NN	O	O
a	NN	O	O
response	NN	O	O
to	NN	O	O
acidic	NN	O	B-protein
activators	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
during	NN	O	O
induction	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
work	NN	O	O
has	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
type	NN	O	B-protein
1	NN	O	I-protein
and	NN	O	I-protein
2A	NN	O	I-protein
protein	NN	O	I-protein
phosphatases	NN	O	I-protein
,	NN	O	O
on	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
U-937	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
treatment	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
differentiated	NN	O	O
monocyte	NN	O	O
phenotype	NN	O	O
characterized	NN	O	O
by	NN	O	O
:	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
growth	NN	O	O
arrest	NN	O	O
;	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
increases	NN	O	O
in	NN	O	O
Mac-1	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
antigen	NN	O	I-protein
expression	NN	O	O
;	NN	O	O
(	NN	O	O
c	NN	O	O
)	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
c-myc	NN	O	B-protein
transcripts	NN	O	I-protein
;	NN	O	O
and	NN	O	O
(	NN	O	O
d	NN	O	O
)	NN	O	O
induction	NN	O	O
of	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

This	NN	O	O
induction	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
transient	NN	O	O
increases	NN	O	O
in	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
,	NN	O	O
which	NN	O	O
were	NN	O	O
maximal	NN	O	O
at	NN	O	O
6	NN	O	O
h	NN	O	O
.	NN	O	O

Similar	NN	O	O
effects	NN	O	O
were	NN	O	O
obtained	NN	O	O
for	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Run-on	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
detectable	NN	O	O
levels	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
transcription	NN	O	O
in	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
that	NN	O	O
this	NN	O	O
rate	NN	O	O
is	NN	O	O
increased	NN	O	O
approximately	NN	O	O
40-fold	NN	O	O
following	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
exposure	NN	O	O
.	NN	O	O

c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
were	NN	O	O
superinduced	NN	O	O
in	NN	O	O
cells	NN	O	O
treated	NN	O	O
with	NN	O	O
both	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
and	NN	O	O
cycloheximide	NN	O	O
,	NN	O	O
whereas	NN	O	O
inhibition	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
had	NN	O	O
little	NN	O	O
,	NN	O	O
if	NN	O	O
any	NN	O	O
,	NN	O	O
effect	NN	O	O
on	NN	O	O
okadaic	NN	O	O
acid-induced	NN	O	O
c-jun	NN	O	B-DNA
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
half-life	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
similar	NN	O	O
(	NN	O	O
45-50	NN	O	O
min	NN	O	O
)	NN	O	O
in	NN	O	O
both	NN	O	O
untreated	NN	O	O
and	NN	O	O
okadaic	NN	O	B-cell_line
acid-induced	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
treatment	NN	O	O
with	NN	O	O
both	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
and	NN	O	O
cycloheximide	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
stabilization	NN	O	O
(	NN	O	O
t	NN	O	O
1/2	NN	O	O
=	NN	O	O
90	NN	O	O
min	NN	O	O
)	NN	O	O
of	NN	O	O
c-jun	NN	O	B-protein
transcripts	NN	O	I-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
transcription	NN	O	O
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
is	NN	O	O
controlled	NN	O	O
primarily	NN	O	O
by	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Since	NN	O	O
previous	NN	O	O
studies	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
c-jun	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
autoinduced	NN	O	O
by	NN	O	O
Jun/AP-1	NN	O	B-protein
,	NN	O	O
we	NN	O	O
also	NN	O	O
studied	NN	O	O
transcription	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	I-DNA
positions	NN	O	I-DNA
-132/+170	NN	O	I-DNA
)	NN	O	I-DNA
-reporter	NN	O	I-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
with	NN	O	O
and	NN	O	O
without	NN	O	O
a	NN	O	O
mutated	NN	O	O
AP-1	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Mitogen	NN	O	O
stimulation	NN	O	O
of	NN	O	O
T-cells	NN	O	O
increases	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
,	NN	O	O
AP-1	NN	O	O
binding	NN	O	O
and	NN	O	O
AP-1	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analysed	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
mitogenic	NN	O	O
lectins	NN	O	O
on	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
on	NN	O	O
activator	NN	O	O
protein-1	NN	O	O
(	NN	O	O
AP-1	NN	O	O
)	NN	O	O
binding	NN	O	O
and	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
in	NN	O	O
Jurkat	NN	O	O
T-cells	NN	O	O
.	NN	O	O

Both	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
were	NN	O	O
increased	NN	O	O
after	NN	O	O
Con	NN	O	O
A	NN	O	O
and	NN	O	O
PHA	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Since	NN	O	O
T-cell	NN	O	O
stimulation	NN	O	O
increases	NN	O	O
both	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
and	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
and	NN	O	O
activates	NN	O	O
protein	NN	O	O
kinase	NN	O	O
C	NN	O	O
(	NN	O	O
PKC	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
possible	NN	O	O
involvement	NN	O	O
of	NN	O	O
these	NN	O	O
intracellular	NN	O	O
messengers	NN	O	O
in	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
induction	NN	O	O
was	NN	O	O
tested	NN	O	O
.	NN	O	O

PMA	NN	O	O
,	NN	O	O
which	NN	O	O
directly	NN	O	O
activates	NN	O	O
PKC	NN	O	O
,	NN	O	O
mimicked	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
lectins	NN	O	O
on	NN	O	O
c-Fos	NN	O	O
and	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
but	NN	O	O
elevation	NN	O	O
of	NN	O	O
either	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
or	NN	O	O
cAMP	NN	O	O
levels	NN	O	O
had	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

The	NN	O	O
mitogen-induced	NN	O	O
increase	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
immunoreactivity	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
H-7	NN	O	O
,	NN	O	O
a	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
with	NN	O	O
relatively	NN	O	O
high	NN	O	O
specificity	NN	O	O
for	NN	O	O
PKC	NN	O	O
,	NN	O	O
and	NN	O	O
less	NN	O	O
efficiently	NN	O	O
by	NN	O	O
H-8	NN	O	O
,	NN	O	O
a	NN	O	O
structurally	NN	O	O
related	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
less	NN	O	O
active	NN	O	O
on	NN	O	O
PKC	NN	O	O
,	NN	O	O
but	NN	O	O
more	NN	O	O
active	NN	O	O
on	NN	O	O
cyclic	NN	O	O
nucleotide-dependent	NN	O	O
kinases	NN	O	O
.	NN	O	O

Con	NN	O	O
A	NN	O	O
stimulation	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
increase	NN	O	O
both	NN	O	O
binding	NN	O	O
of	NN	O	O
AP-1	NN	O	O
to	NN	O	O
the	NN	O	O
AP-1	NN	O	O
consensus	NN	O	O
sequence	NN	O	O
,	NN	O	O
TRE	NN	O	O
,	NN	O	O
and	NN	O	O
AP-1	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
,	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

PMA	NN	O	O
was	NN	O	O
also	NN	O	O
found	NN	O	O
to	NN	O	O
increase	NN	O	O
the	NN	O	O
AP-1	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
,	NN	O	O
whereas	NN	O	O
elevation	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
or	NN	O	O
cAMP	NN	O	O
had	NN	O	O
only	NN	O	O
minor	NN	O	O
effects	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
stimulation	NN	O	O
with	NN	O	O
mitogenic	NN	O	O
lectins	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
increase	NN	O	O
both	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
c-Jun	NN	O	B-protein
protein	NN	O	O
levels	NN	O	O
,	NN	O	O
AP-1	NN	O	O
binding	NN	O	O
and	NN	O	O
AP-1	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
that	NN	O	O
they	NN	O	O
act	NN	O	O
via	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
could	NN	O	O
involve	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
11-base-pair	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
motif	NN	O	O
apparently	NN	O	O
unique	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin	NN	O	I-DNA
4	NN	O	I-DNA
gene	NN	O	I-DNA
confers	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
DNA	NN	O	B-DNA
segment	NN	O	I-DNA
that	NN	O	O
confers	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
signals	NN	O	O
on	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin	NN	O	I-DNA
(	NN	O	I-DNA
IL	NN	O	I-DNA
)	NN	O	I-DNA
4	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
of	NN	O	O
10	NN	O	O
kilobases	NN	O	O
,	NN	O	O
is	NN	O	O
composed	NN	O	O
of	NN	O	O
four	NN	O	O
exons	NN	O	B-DNA
and	NN	O	O
three	NN	O	O
introns	NN	O	B-DNA
.	NN	O	O

A	NN	O	O
cis-acting	NN	O	B-DNA
element	NN	O	I-DNA
(	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
)	NN	O	O
resides	NN	O	O
in	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
upstream	NN	O	I-DNA
region	NN	O	I-DNA
;	NN	O	O
no	NN	O	O
additional	NN	O	O
DNA	NN	O	B-DNA
segments	NN	O	I-DNA
with	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
were	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

For	NN	O	O
further	NN	O	O
mapping	NN	O	O
purposes	NN	O	O
,	NN	O	O
a	NN	O	O
fusion	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
constructed	NN	O	O
with	NN	O	O
the	NN	O	O
granulocyte/macrophage	NN	O	B-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
basic	NN	O	I-DNA
promoter	NN	O	I-DNA
containing	NN	O	O
60	NN	O	B-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
of	NN	O	I-DNA
sequence	NN	O	I-DNA
upstream	NN	O	O
from	NN	O	O
the	NN	O	O
cap	NN	O	B-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
granulocyte/macrophage	NN	O	I-DNA
colony-stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
various	NN	O	O
lengths	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
upstream	NN	O	I-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
was	NN	O	O
located	NN	O	O
between	NN	O	O
positions	NN	O	O
-79	NN	O	O
and	NN	O	O
-69	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
this	NN	O	O
location	NN	O	O
was	NN	O	O
confirmed	NN	O	O
by	NN	O	O
base-substitution	NN	O	O
mutations	NN	O	O
.	NN	O	O

The	NN	O	O
plasmids	NN	O	O
carrying	NN	O	O
multiple	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
showed	NN	O	O
higher	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
the	NN	O	O
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
that	NN	O	O
recognize	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
by	NN	O	O
DNA-mobility-shift	NN	O	O
assays	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
(	NN	O	O
a	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
that	NN	O	O
specifically	NN	O	O
recognizes	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
)	NN	O	O
to	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
was	NN	O	O
abolished	NN	O	O
when	NN	O	O
oligonucleotides	NN	O	O
carrying	NN	O	O
base	NN	O	O
substitutions	NN	O	O
were	NN	O	O
used	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
(	NN	O	I-protein
P	NN	O	I-protein
)	NN	O	I-protein
interaction	NN	O	O
is	NN	O	O
sequence-specific	NN	O	O
and	NN	O	O
that	NN	O	O
binding	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
paralleled	NN	O	O
the	NN	O	O
sequence	NN	O	O
requirements	NN	O	O
for	NN	O	O
IL-4	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

The	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
does	NN	O	O
not	NN	O	O
share	NN	O	O
homology	NN	O	O
with	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
upstream	NN	O	I-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
even	NN	O	O
though	NN	O	O
surrounding	NN	O	O
sequences	NN	O	O
of	NN	O	O
the	NN	O	O
IL-4	NN	O	B-DNA
gene	NN	O	I-DNA
share	NN	O	O
high	NN	O	O
homology	NN	O	O
with	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
a	NN	O	O
different	NN	O	O
set	NN	O	O
of	NN	O	O
proteins	NN	O	B-protein
recognize	NN	O	O
IL-2	NN	O	B-DNA
and	NN	O	I-DNA
IL-4	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Leukotriene	NN	O	O
B4	NN	O	O
stimulates	NN	O	O
c-fos	NN	O	O
and	NN	O	O
c-jun	NN	O	O
gene	NN	O	O
transcription	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
leukotriene	NN	O	O
B4	NN	O	O
(	NN	O	O
LTB4	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
potent	NN	O	O
lipid	NN	O	O
proinflammatory	NN	O	B-protein
mediator	NN	O	I-protein
,	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
proto-oncogenes	NN	O	B-DNA
c-jun	NN	O	I-DNA
and	NN	O	I-DNA
c-fos	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
we	NN	O	O
looked	NN	O	O
at	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
binding	NN	O	O
specifically	NN	O	O
to	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
element	NN	O	I-DNA
after	NN	O	O
LTB4	NN	O	O
stimulation	NN	O	O
.	NN	O	O

LTB4	NN	O	O
increased	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
time-	NN	O	O
and	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

The	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral-blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
at	NN	O	O
relatively	NN	O	O
high	NN	O	O
levels	NN	O	O
,	NN	O	O
was	NN	O	O
also	NN	O	O
slightly	NN	O	O
augmented	NN	O	O
by	NN	O	O
LTB4	NN	O	O
,	NN	O	O
although	NN	O	O
to	NN	O	O
a	NN	O	O
much	NN	O	O
lower	NN	O	O
extent	NN	O	O
than	NN	O	O
c-fos	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
kinetics	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
genes	NN	O	O
were	NN	O	O
also	NN	O	O
slightly	NN	O	O
different	NN	O	O
,	NN	O	O
with	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
reaching	NN	O	O
a	NN	O	O
peak	NN	O	O
at	NN	O	O
15	NN	O	O
min	NN	O	O
after	NN	O	O
stimulation	NN	O	O
and	NN	O	O
c-jun	NN	O	B-DNA
at	NN	O	O
30	NN	O	O
min	NN	O	O
.	NN	O	O

Both	NN	O	O
messages	NN	O	O
rapidly	NN	O	O
declined	NN	O	O
thereafter	NN	O	O
.	NN	O	O

Stability	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
LTB4	NN	O	O
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
after	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
treatment	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
transcription	NN	O	O
studies	NN	O	O
in	NN	O	O
vitro	NN	O	O
showed	NN	O	O
that	NN	O	O
LTB4	NN	O	O
increased	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
gene	NN	O	I-DNA
7-fold	NN	O	O
and	NN	O	O
the	NN	O	O
c-jun	NN	O	B-DNA
gene	NN	O	I-DNA
1.4-fold	NN	O	O
.	NN	O	O

Resting	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
contained	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
stimulation	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
with	NN	O	O
LTB4	NN	O	O
induced	NN	O	O
greater	NN	O	O
AP-1	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
LTB4	NN	O	O
may	NN	O	O
regulate	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
different	NN	O	O
cytokines	NN	O	B-protein
by	NN	O	O
modulating	NN	O	O
the	NN	O	O
yield	NN	O	O
and/or	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
AP-1	NN	O	B-protein
-binding	NN	O	O
proto-oncogene	NN	O	B-protein
products	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Modulation	NN	O	O
of	NN	O	O
normal	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
by	NN	O	O
the	NN	O	O
endogenous	NN	O	O
thyroid	NN	O	O
hormone	NN	O	O
and	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptors	NN	O	I-protein
:	NN	O	O
a	NN	O	O
possible	NN	O	O
target	NN	O	O
for	NN	O	O
v-erbA	NN	O	B-DNA
oncogene	NN	O	I-DNA
action	NN	O	O
.	NN	O	O

The	NN	O	O
v-erbA	NN	O	B-DNA
oncogene	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
mutated	NN	O	O
version	NN	O	O
of	NN	O	O
the	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
c-erbA/TR-alpha	NN	O	B-protein
)	NN	O	O
,	NN	O	O
inhibits	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
and	NN	O	O
constitutively	NN	O	O
represses	NN	O	O
transcription	NN	O	O
of	NN	O	O
certain	NN	O	O
erythrocyte	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
normal	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
proto-oncogene	NN	O	B-DNA
c-erbA	NN	O	I-DNA
in	NN	O	O
erythropoiesis	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
endogenous	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
c-erbA/TR-alpha	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
closely	NN	O	O
related	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
RAR-alpha	NN	O	B-protein
)	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
normal	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Retinoic	NN	O	O
acid	NN	O	O
(	NN	O	O
RA	NN	O	O
)	NN	O	O
distinctly	NN	O	O
modulated	NN	O	O
the	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
program	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
and	NN	O	O
erythroblasts	NN	O	O
reversibly	NN	O	O
transformed	NN	O	O
by	NN	O	O
a	NN	O	O
conditional	NN	O	O
tyrosine	NN	O	B-DNA
kinase	NN	O	I-DNA
oncogene	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
added	NN	O	O
pulsewise	NN	O	O
to	NN	O	O
immature	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
differentiation	NN	O	O
was	NN	O	O
accelerated	NN	O	O
while	NN	O	O
more	NN	O	O
mature	NN	O	B-cell_type
cells	NN	O	I-cell_type
underwent	NN	O	O
premature	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

Thyroid	NN	O	O
hormone	NN	O	O
(	NN	O	O
T3	NN	O	O
)	NN	O	O
alone	NN	O	O
caused	NN	O	O
similar	NN	O	O
but	NN	O	O
weaker	NN	O	O
effects	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
T3	NN	O	O
strongly	NN	O	O
enhanced	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
RA	NN	O	O
,	NN	O	O
suggesting	NN	O	O
cooperative	NN	O	O
action	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
receptors	NN	O	O
in	NN	O	O
modulating	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
RAR-alpha	NN	O	I-protein
in	NN	O	O
receptor-negative	NN	O	B-cell_line
erythroblasts	NN	O	I-cell_line
conferred	NN	O	O
RA-induced	NN	O	O
regulation	NN	O	O
of	NN	O	O
differentiation	NN	O	O
to	NN	O	O
the	NN	O	O
otherwise	NN	O	O
unresponsive	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
thus	NN	O	O
showing	NN	O	O
that	NN	O	O
the	NN	O	O
RAR-alpha	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
RA	NN	O	O
effect	NN	O	O
.	NN	O	O

Likewise	NN	O	O
,	NN	O	O
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
exogenous	NN	O	B-protein
c-erbA/TR-alpha	NN	O	I-protein
in	NN	O	O
erythroblasts	NN	O	B-cell_type
rendered	NN	O	O
them	NN	O	O
susceptible	NN	O	O
to	NN	O	O
modulation	NN	O	O
of	NN	O	O
differentiation	NN	O	O
by	NN	O	O
T3	NN	O	O
,	NN	O	O
suggesting	NN	O	O
a	NN	O	O
similar	NN	O	O
function	NN	O	O
of	NN	O	O
both	NN	O	O
receptors	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
lymphokine	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
role	NN	O	O
of	NN	O	O
cis-acting	NN	O	B-DNA
DNA	NN	O	I-DNA
elements	NN	O	I-DNA
that	NN	O	O
respond	NN	O	O
to	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
initiated	NN	O	O
by	NN	O	O
the	NN	O	O
recognition	NN	O	O
of	NN	O	O
antigen	NN	O	O
on	NN	O	O
antigen	NN	O	B-cell_type
presenting	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
exert	NN	O	O
the	NN	O	O
effector	NN	O	O
functions	NN	O	O
in	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

Two	NN	O	O
types	NN	O	O
of	NN	O	O
helper	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
(	NN	O	I-cell_line
Th	NN	O	I-cell_line
)	NN	O	I-cell_line
clones	NN	O	I-cell_line
(	NN	O	O
Th1	NN	O	B-cell_line
and	NN	O	O
Th2	NN	O	B-cell_line
)	NN	O	O
are	NN	O	O
defined	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
different	NN	O	O
patterns	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
(	NN	O	O
lymphokine	NN	O	B-protein
)	NN	O	O
secretion	NN	O	O
.	NN	O	O

They	NN	O	O
determine	NN	O	O
the	NN	O	O
outcome	NN	O	O
of	NN	O	O
an	NN	O	O
antigenic	NN	O	O
response	NN	O	O
toward	NN	O	O
humoral	NN	O	O
or	NN	O	O
cell-mediated	NN	O	O
immunity	NN	O	O
.	NN	O	O

Although	NN	O	O
lymphokine	NN	O	B-DNA
genes	NN	O	I-DNA
are	NN	O	O
coordinately	NN	O	O
regulated	NN	O	O
upon	NN	O	O
antigen	NN	O	O
stimulation	NN	O	O
,	NN	O	O
they	NN	O	O
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
common	NN	O	O
to	NN	O	O
all	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
those	NN	O	O
which	NN	O	O
are	NN	O	O
unique	NN	O	O
to	NN	O	O
each	NN	O	O
gene	NN	O	O
.	NN	O	O

For	NN	O	O
most	NN	O	O
lymphokine	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
(	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13	NN	O	O
acetate	NN	O	O
,	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophores	NN	O	O
(	NN	O	O
A23187	NN	O	O
)	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
their	NN	O	O
maximal	NN	O	O
induction	NN	O	O
.	NN	O	O

Yet	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
alone	NN	O	O
or	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
alone	NN	O	O
produce	NN	O	O
several	NN	O	O
lymphokines	NN	O	B-protein
.	NN	O	O

The	NN	O	O
production	NN	O	O
of	NN	O	O
the	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony	NN	O	I-protein
stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
is	NN	O	O
completely	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
two	NN	O	O
signals	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
found	NN	O	O
a	NN	O	O
cis-acting	NN	O	B-DNA
region	NN	O	I-DNA
spanning	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
positions	NN	O	O
-95	NN	O	O
to	NN	O	O
+27	NN	O	O
)	NN	O	O
that	NN	O	O
confers	NN	O	O
inducibility	NN	O	O
to	NN	O	O
reporter	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
.	NN	O	O

Further	NN	O	O
analysis	NN	O	O
identified	NN	O	O
three	NN	O	O
elements	NN	O	O
required	NN	O	O
for	NN	O	O
efficient	NN	O	O
induction	NN	O	O
,	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
GM2	NN	O	B-DNA
,	NN	O	O
GC-box	NN	O	B-DNA
and	NN	O	O
conserved	NN	O	B-DNA
lymphokine	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
CLE0	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

GM2	NN	O	B-DNA
defines	NN	O	O
a	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
protein	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
whose	NN	O	O
binding	NN	O	O
is	NN	O	O
inducible	NN	O	O
by	NN	O	O
PMA	NN	O	O
.	NN	O	O

One	NN	O	O
protein	NN	O	O
,	NN	O	O
NF-GM2	NN	O	B-protein
is	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kB	NN	O	B-protein
.	NN	O	O

GC-box	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
constitutively	NN	O	B-protein
bound	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

CLEO	NN	O	O
defines	NN	O	O
a	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
protein	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
whose	NN	O	O
optimum	NN	O	O
binding	NN	O	O
is	NN	O	O
stimulated	NN	O	O
by	NN	O	O
PMA	NN	O	O
and	NN	O	O
A23187	NN	O	O
.	NN	O	O

Viral	NN	O	B-protein
trans-activators	NN	O	I-protein
such	NN	O	O
as	NN	O	O
Tax	NN	O	B-protein
(	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
virus-1	NN	O	O
,	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
and	NN	O	O
E2	NN	O	B-protein
(	NN	O	O
bovine	NN	O	O
papilloma	NN	O	O
virus	NN	O	O
,	NN	O	O
BPV	NN	O	O
)	NN	O	O
proteins	NN	O	O
are	NN	O	O
other	NN	O	O
agents	NN	O	O
which	NN	O	O
activate	NN	O	O
lymphokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
bypassing	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
mediated	NN	O	O
signaling	NN	O	O
.	NN	O	O

The	NN	O	O
trans-activation	NN	O	O
domain	NN	O	O
of	NN	O	O
E2	NN	O	B-protein
and	NN	O	O
Tax	NN	O	B-protein
is	NN	O	O
interchangeable	NN	O	O
although	NN	O	O
they	NN	O	O
have	NN	O	O
no	NN	O	O
obvious	NN	O	O
sequence	NN	O	O
homology	NN	O	O
between	NN	O	O
them	NN	O	O
.	NN	O	O

The	NN	O	O
viral	NN	O	B-protein
trans-activators	NN	O	I-protein
appear	NN	O	O
to	NN	O	O
target	NN	O	O
specific	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
such	NN	O	O
as	NN	O	O
NF-kB	NN	O	B-protein
and	NN	O	O
Sp1	NN	O	B-protein
to	NN	O	O
cis-acting	NN	O	B-DNA
DNA	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
promote	NN	O	O
lymphokine	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
without	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
stimulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Pit-1	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
children	NN	O	O
with	NN	O	O
combined	NN	O	O
pituitary	NN	O	O
hormone	NN	O	O
deficiency	NN	O	O
.	NN	O	O

Pit-1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
pituitary-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
binds	NN	O	O
to	NN	O	O
and	NN	O	O
transactivates	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
growth	NN	O	B-DNA
hormone	NN	O	I-DNA
and	NN	O	I-DNA
prolactin	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
three	NN	O	O
unrelated	NN	O	O
Japanese	NN	O	O
children	NN	O	O
with	NN	O	O
combined	NN	O	O
pituitary	NN	O	O
hormone	NN	O	O
deficiency	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
three	NN	O	O
point	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
Pit-1	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
Pro24Leu	NN	O	B-DNA
,	NN	O	O
Arg143Gln	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
Arg271Trp	NN	O	B-DNA
,	NN	O	O
located	NN	O	O
on	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
transactivation	NN	O	I-DNA
region	NN	O	I-DNA
,	NN	O	O
POU-specific	NN	O	B-DNA
domain	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
POU-homeodomain	NN	O	B-DNA
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
and	NN	O	O
elevation	NN	O	O
of	NN	O	O
cAMP	NN	O	O
interact	NN	O	O
synergistically	NN	O	O
to	NN	O	O
raise	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
Jurkat	NN	O	O
cells	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
earlier	NN	O	O
found	NN	O	O
that	NN	O	O
in	NN	O	O
Jurkat	NN	O	O
cells	NN	O	O
activation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
enhances	NN	O	O
the	NN	O	O
cyclic	NN	O	O
adenosine	NN	O	O
monophosphate	NN	O	O
(	NN	O	O
cAMP	NN	O	O
)	NN	O	O
accumulation	NN	O	O
induced	NN	O	O
by	NN	O	O
adenosine	NN	O	O
receptor	NN	O	O
stimulation	NN	O	O
or	NN	O	O
activation	NN	O	O
of	NN	O	O
Gs	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
have	NN	O	O
therefore	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
PMA	NN	O	O
(	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
)	NN	O	O
which	NN	O	O
stimulates	NN	O	O
PKC	NN	O	B-protein
and	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
the	NN	O	O
adenosine	NN	O	B-protein
receptor	NN	O	I-protein
agonist	NN	O	O
NECA	NN	O	O
(	NN	O	O
5'-	NN	O	O
(	NN	O	O
N-ethyl	NN	O	O
)	NN	O	O
-carboxamido	NN	O	O
adenosine	NN	O	O
)	NN	O	O
and	NN	O	O
forskolin	NN	O	O
to	NN	O	O
raise	NN	O	O
cAMP	NN	O	O
,	NN	O	O
on	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
and	NN	O	O
on	NN	O	O
the	NN	O	O
binding	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
activator	NN	O	B-protein
protein-1	NN	O	I-protein
(	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
.	NN	O	O

PMA	NN	O	O
treatment	NN	O	O
caused	NN	O	O
a	NN	O	O
concentration-	NN	O	O
and	NN	O	O
time-dependent	NN	O	O
increase	NN	O	O
in	NN	O	O
both	NN	O	O
c-Fos	NN	O	O
and	NN	O	O
Jun	NN	O	O
immunoreactivity	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
cAMP	NN	O	O
elevation	NN	O	O
that	NN	O	O
had	NN	O	O
only	NN	O	O
a	NN	O	O
slight	NN	O	O
effect	NN	O	O
.	NN	O	O

Both	NN	O	O
PMA	NN	O	O
and	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
NECA	NN	O	O
and	NN	O	O
forskolin	NN	O	O
acted	NN	O	O
together	NN	O	O
either	NN	O	O
to	NN	O	O
increase	NN	O	O
(	NN	O	O
c-Fos	NN	O	B-protein
)	NN	O	O
or	NN	O	O
decrease	NN	O	O
(	NN	O	O
Jun	NN	O	B-protein
)	NN	O	O
protein	NN	O	O
levels	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
increasing	NN	O	O
AP-1	NN	O	B-protein
binding	NN	O	O
,	NN	O	O
as	NN	O	O
judged	NN	O	O
by	NN	O	O
gel-shift	NN	O	O
assay	NN	O	O
,	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
clear-cut	NN	O	O
synergy	NN	O	O
between	NN	O	O
the	NN	O	O
PKC	NN	O	B-protein
stimulator	NN	O	O
and	NN	O	O
the	NN	O	O
cAMP	NN	O	O
elevating	NN	O	O
agents	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
simultaneous	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
and	NN	O	O
elevation	NN	O	O
of	NN	O	O
cAMP	NN	O	O
leads	NN	O	O
to	NN	O	O
an	NN	O	O
enhanced	NN	O	O
AP-1	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
T-leukemia	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
previously	NN	O	O
observed	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
parallel	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
may	NN	O	O
have	NN	O	O
functional	NN	O	O
consequences	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
anti-CD3	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
-induced	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
isoproterenol	NN	O	O
,	NN	O	O
or	NN	O	O
prostaglandin	NN	O	O
E2	NN	O	O
either	NN	O	O
alone	NN	O	O
or	NN	O	O
in	NN	O	O
combination	NN	O	O
.	NN	O	O

1	NN	O	O
.	NN	O	O

The	NN	O	O
purpose	NN	O	O
of	NN	O	O
these	NN	O	O
studies	NN	O	O
was	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
proliferation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
obtained	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
DEX	NN	O	O
)	NN	O	O
,	NN	O	O
isoproterenol	NN	O	O
(	NN	O	O
ISO	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
prostaglandin	NN	O	O
E2	NN	O	O
(	NN	O	O
PGE2	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
former	NN	O	O
two	NN	O	O
substances	NN	O	O
interact	NN	O	O
with	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
and	NN	O	I-protein
beta-adrenergic	NN	O	I-protein
receptors	NN	O	I-protein
respectively	NN	O	O
.	NN	O	O

When	NN	O	O
occupied	NN	O	O
by	NN	O	O
their	NN	O	O
natural	NN	O	O
ligands	NN	O	O
,	NN	O	O
glucocorticosteroids	NN	O	O
and	NN	O	O
catecholamines	NN	O	O
,	NN	O	O
these	NN	O	O
receptors	NN	O	O
have	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
modulating	NN	O	O
T-cell	NN	O	O
function	NN	O	O
during	NN	O	O
stress	NN	O	O
.	NN	O	O

During	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
PGE2	NN	O	O
bind	NN	O	O
to	NN	O	O
their	NN	O	O
receptors	NN	O	O
on	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
thus	NN	O	O
alter	NN	O	O
responsiveness	NN	O	O
.	NN	O	O

Proliferation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
induced	NN	O	O
by	NN	O	O
immobilized	NN	O	O
anti-CD3	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
(	NN	O	O
mAb	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
an	NN	O	O
additional	NN	O	O
costimulatory	NN	O	O
signal	NN	O	O
delivered	NN	O	O
by	NN	O	O
anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
.	NN	O	O

2	NN	O	O
.	NN	O	O

Various	NN	O	O
physiologic	NN	O	O
concentrations	NN	O	O
of	NN	O	O
DEX	NN	O	O
,	NN	O	O
ISO	NN	O	O
,	NN	O	O
or	NN	O	O
PGE2	NN	O	O
were	NN	O	O
added	NN	O	O
at	NN	O	O
the	NN	O	O
time	NN	O	O
of	NN	O	O
initiation	NN	O	O
of	NN	O	O
the	NN	O	O
cultures	NN	O	O
and	NN	O	O
subsequent	NN	O	O
proliferation	NN	O	O
of	NN	O	O
the	NN	O	O
unstimulated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
determined	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
physiologic	NN	O	O
concentrations	NN	O	O
of	NN	O	O
all	NN	O	O
three	NN	O	O
of	NN	O	O
these	NN	O	O
agents	NN	O	O
inhibit	NN	O	O
the	NN	O	O
anti-CD3	NN	O	B-protein
mAb	NN	O	I-protein
-induced	NN	O	O
proliferation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

3	NN	O	O
.	NN	O	O

Although	NN	O	O
DEX	NN	O	O
and	NN	O	O
PGE2	NN	O	O
were	NN	O	O
equipotent	NN	O	O
in	NN	O	O
suppressing	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
ISO	NN	O	O
was	NN	O	O
much	NN	O	O
less	NN	O	O
effective	NN	O	O
.	NN	O	O

4	NN	O	O
.	NN	O	O

Because	NN	O	O
concomitant	NN	O	O
elevations	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
levels	NN	O	O
of	NN	O	O
these	NN	O	O
substances	NN	O	O
may	NN	O	O
occur	NN	O	O
,	NN	O	O
experiments	NN	O	O
were	NN	O	O
performed	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
T-cell	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
DEX	NN	O	O
together	NN	O	O
with	NN	O	O
either	NN	O	O
PGE2	NN	O	O
or	NN	O	O
ISO	NN	O	O
.	NN	O	O

Synergistic	NN	O	O
suppression	NN	O	O
of	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
was	NN	O	O
observed	NN	O	O
when	NN	O	O
various	NN	O	O
concentrations	NN	O	O
of	NN	O	O
DEX	NN	O	O
and	NN	O	O
PGE2	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
DEX	NN	O	O
and	NN	O	O
ISO	NN	O	O
,	NN	O	O
were	NN	O	O
added	NN	O	O
to	NN	O	O
cultures	NN	O	O
.	NN	O	O

This	NN	O	O
synergistic	NN	O	O
suppression	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
cAMP	NN	O	O
accumulation	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
with	NN	O	O
DEX	NN	O	O
and	NN	O	O
PGE2	NN	O	O
.	NN	O	O

5	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
anti-CD28	NN	O	B-protein
mAb	NN	O	I-protein
to	NN	O	O
anti-CD3	NN	O	B-cell_type
mAb-stimulated	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
overcame	NN	O	O
much	NN	O	O
of	NN	O	O
the	NN	O	O
suppression	NN	O	O
of	NN	O	O
proliferation	NN	O	O
induced	NN	O	O
by	NN	O	O
PGE2	NN	O	O
or	NN	O	O
ISO	NN	O	O
but	NN	O	O
less	NN	O	O
so	NN	O	O
than	NN	O	O
that	NN	O	O
induced	NN	O	O
by	NN	O	O
DEX	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
footprint	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
HLA-DRA	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
:	NN	O	O
cell-specific	NN	O	O
interaction	NN	O	O
at	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
and	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
X	NN	O	O
box	NN	O	O
binding	NN	O	O
by	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
complex	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
DRA	NN	O	B-DNA
has	NN	O	O
previously	NN	O	O
identified	NN	O	O
at	NN	O	O
least	NN	O	O
five	NN	O	O
cis-acting	NN	O	B-DNA
regions	NN	O	I-DNA
required	NN	O	O
for	NN	O	O
maximal	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
for	NN	O	O
protein-DNA	NN	O	O
interactions	NN	O	O
in	NN	O	O
the	NN	O	O
intact	NN	O	B-cell_type
cell	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
may	NN	O	O
mediate	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

Using	NN	O	O
in	NN	O	O
vivo	NN	O	O
genomic	NN	O	O
footprinting	NN	O	O
we	NN	O	O
identified	NN	O	O
interactions	NN	O	O
in	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
at	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
Y	NN	O	B-DNA
,	NN	O	I-DNA
X1	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
X2	NN	O	I-DNA
boxes	NN	O	I-DNA
.	NN	O	O

Class	NN	O	B-cell_line
II	NN	O	I-cell_line
antigen	NN	O	I-cell_line
expressing	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
maintained	NN	O	O
contacts	NN	O	O
identical	NN	O	O
to	NN	O	O
B-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
class	NN	O	B-cell_line
II-negative	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
exhibited	NN	O	O
no	NN	O	O
interactions	NN	O	O
.	NN	O	O

In	NN	O	O
lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
occupied	NN	O	O
and	NN	O	O
required	NN	O	O
for	NN	O	O
maximal	NN	O	O
expression	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
most	NN	O	O
likely	NN	O	O
due	NN	O	O
to	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
lymphoid-specific	NN	O	B-protein
OTF-2	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
class	NN	O	B-cell_line
II-positive	NN	O	I-cell_line
nonlymphoid	NN	O	I-cell_line
glioblastoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
does	NN	O	O
not	NN	O	O
exhibit	NN	O	O
interactions	NN	O	O
at	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
despite	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
ubiquitous	NN	O	B-protein
OTF-1	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
an	NN	O	O
open	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
discriminates	NN	O	O
against	NN	O	O
OTF-1	NN	O	B-protein
activation	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
in	NN	O	O
the	NN	O	O
glioblastoma	NN	O	B-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Interferon	NN	O	B-protein
gamma	NN	O	I-protein
induces	NN	O	O
class	NN	O	O
II	NN	O	O
expression	NN	O	O
in	NN	O	O
this	NN	O	O
glioblastoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
,	NN	O	O
in	NN	O	O
parallel	NN	O	O
,	NN	O	O
up-regulates	NN	O	O
X1	NN	O	O
and	NN	O	O
X2	NN	O	O
box	NN	O	O
protein-DNA	NN	O	O
interactions	NN	O	O
,	NN	O	O
while	NN	O	O
all	NN	O	O
other	NN	O	O
interactions	NN	O	O
remain	NN	O	O
unchanged	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
functions	NN	O	O
on	NN	O	O
a	NN	O	O
poised	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
altering	NN	O	O
weak	NN	O	O
,	NN	O	O
nonproductive	NN	O	O
interactions	NN	O	O
at	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
boxes	NN	O	I-DNA
to	NN	O	O
strong	NN	O	O
interactions	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
provide	NN	O	O
direct	NN	O	O
in	NN	O	O
vivo	NN	O	O
evidence	NN	O	O
to	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
modulation	NN	O	O
of	NN	O	O
X1	NN	O	B-DNA
and	NN	O	O
X2	NN	O	B-DNA
interactions	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
constituent	NN	O	O
of	NN	O	O
the	NN	O	O
interferon	NN	O	B-protein
gamma	NN	O	I-protein
induction	NN	O	O
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	O

Estrogen	NN	O	O
binding	NN	O	O
sites	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
effects	NN	O	O
of	NN	O	O
danazol	NN	O	O
on	NN	O	O
their	NN	O	O
sites	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

1	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
was	NN	O	O
designed	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
estrogen	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
high	NN	O	O
affinity	NN	O	O
,	NN	O	O
low	NN	O	O
capacity	NN	O	O
)	NN	O	O
and	NN	O	O
type	NN	O	O
II	NN	O	O
(	NN	O	O
low	NN	O	O
affinity	NN	O	O
,	NN	O	O
high	NN	O	O
capacity	NN	O	O
)	NN	O	O
binding	NN	O	O
sites	NN	O	O
in	NN	O	O
human	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
danazol	NN	O	O
on	NN	O	O
these	NN	O	O
sites	NN	O	O
.	NN	O	O

2	NN	O	O
.	NN	O	O

These	NN	O	O
two	NN	O	O
types	NN	O	O
of	NN	O	O
estrogen	NN	O	O
binding	NN	O	O
sites	NN	O	O
existed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

3	NN	O	O
.	NN	O	O

Danazol	NN	O	O
bound	NN	O	O
to	NN	O	O
these	NN	O	O
sites	NN	O	O
in	NN	O	O
high	NN	O	O
concentration	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
M	NN	O	O
,	NN	O	O
clinical	NN	O	O
serum	NN	O	O
concentration	NN	O	O
during	NN	O	O
danazol	NN	O	O
therapy	NN	O	O
)	NN	O	O
and	NN	O	O
decreased	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
both	NN	O	O
sites	NN	O	O
.	NN	O	O

4	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
suggested	NN	O	O
that	NN	O	O
danazol	NN	O	O
has	NN	O	O
an	NN	O	O
anti-estrogenic	NN	O	O
action	NN	O	O
to	NN	O	O
the	NN	O	O
monocytes	NN	O	O
through	NN	O	O
the	NN	O	O
competition	NN	O	O
and	NN	O	O
suppression	NN	O	O
of	NN	O	O
estrogen	NN	O	O
binding	NN	O	O
sites	NN	O	O
as	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
estrogen	NN	O	O
target	NN	O	O
organ	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
microtitre	NN	O	O
assay	NN	O	O
system	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
:	NN	O	O
decreased	NN	O	O
receptor	NN	O	O
concentration	NN	O	O
in	NN	O	O
myocardial	NN	O	O
infarction	NN	O	O
.	NN	O	O

A	NN	O	O
major	NN	O	O
difficulty	NN	O	O
in	NN	O	O
determination	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
sites	NN	O	O
is	NN	O	O
the	NN	O	O
very	NN	O	O
complicated	NN	O	O
assay	NN	O	O
procedure	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
a	NN	O	O
microtitre	NN	O	O
assay	NN	O	O
system	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
which	NN	O	O
is	NN	O	O
a	NN	O	O
whole-cell	NN	O	O
competitive	NN	O	O
binding	NN	O	O
radioassay	NN	O	O
using	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-dexamethasone	NN	O	O
as	NN	O	O
radioligand	NN	O	O
.	NN	O	O

This	NN	O	O
modification	NN	O	O
of	NN	O	O
a	NN	O	O
previously	NN	O	O
described	NN	O	O
protocol	NN	O	O
simplifies	NN	O	O
and	NN	O	O
reduces	NN	O	O
laboratory	NN	O	O
work	NN	O	O
and	NN	O	O
allows	NN	O	O
assay	NN	O	O
reproducibility	NN	O	O
to	NN	O	O
be	NN	O	O
controlled	NN	O	O
more	NN	O	O
reliably	NN	O	O
.	NN	O	O

Thus	NN	O	O
enabled	NN	O	O
to	NN	O	O
perform	NN	O	O
the	NN	O	O
test	NN	O	O
on	NN	O	O
multiple	NN	O	O
blood	NN	O	B-cell_type
samples	NN	O	I-cell_type
in	NN	O	O
parallel	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
cardiac	NN	O	O
infarction	NN	O	O
patients	NN	O	O
over	NN	O	O
a	NN	O	O
12-day	NN	O	O
period	NN	O	O
to	NN	O	O
test	NN	O	O
if	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
is	NN	O	O
altered	NN	O	O
in	NN	O	O
this	NN	O	O
'stressful	NN	O	O
'	NN	O	O
disease	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
first	NN	O	O
day	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
,	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
capacity	NN	O	O
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
without	NN	O	O
alteration	NN	O	O
of	NN	O	O
the	NN	O	O
receptor-ligand	NN	O	O
affinity	NN	O	O
,	NN	O	O
whereas	NN	O	O
on	NN	O	O
days	NN	O	O
4	NN	O	O
and	NN	O	O
12	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
receptor	NN	O	O
sites	NN	O	O
was	NN	O	O
normal	NN	O	O
again	NN	O	O
.	NN	O	O

This	NN	O	O
result	NN	O	O
fits	NN	O	O
well	NN	O	O
into	NN	O	O
the	NN	O	O
general	NN	O	O
observation	NN	O	O
of	NN	O	O
stress-induced	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
immune	NN	O	O
responses	NN	O	O
.	NN	O	O

-DOCSTART-	O

SRC-related	NN	O	B-DNA
proto-oncogenes	NN	O	I-DNA
and	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
primary	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
modulation	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
induces	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
encoding	NN	O	O
for	NN	O	O
lymphokines	NN	O	B-protein
.	NN	O	O

Interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
is	NN	O	O
controlled	NN	O	O
transcriptionally	NN	O	O
by	NN	O	O
the	NN	O	O
cooperative	NN	O	O
activity	NN	O	O
of	NN	O	O
specific	NN	O	O
trans-activating	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
and	NN	O	O
FK506	NN	O	O
inhibit	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

A	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
src	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
lymphocyte-specific	NN	O	B-protein
protein	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
p56lck	NN	O	B-protein
,	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

CsA	NN	O	O
was	NN	O	O
found	NN	O	O
not	NN	O	O
to	NN	O	O
inhibit	NN	O	O
lck	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
nor	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
lck	NN	O	B-DNA
gene	NN	O	I-DNA
product	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
CsA	NN	O	O
and	NN	O	O
FK506	NN	O	O
inhibit	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
factors	NN	O	O
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-DNA
and	NN	O	O
AP-1	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Since	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
is	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
same	NN	O	O
stimuli	NN	O	O
that	NN	O	O
stimulate	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
immunosuppressant	NN	O	O
action	NN	O	O
of	NN	O	O
CsA	NN	O	O
and	NN	O	O
FK506	NN	O	O
is	NN	O	O
exerted	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
these	NN	O	O
trans-activating	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
-150	NN	O	B-DNA
bp	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
represent	NN	O	O
the	NN	O	O
major	NN	O	O
target	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
in	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
antigen	NN	O	O
results	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
several	NN	O	O
kinases	NN	O	B-protein
,	NN	O	O
including	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
,	NN	O	O
that	NN	O	O
may	NN	O	O
mediate	NN	O	O
the	NN	O	O
later	NN	O	O
induction	NN	O	O
of	NN	O	O
activation-related	NN	O	O
genes	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
in	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
through	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor/CD3	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
prolonged	NN	O	O
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
untransformed	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
Ar-5	NN	O	I-cell_line
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
results	NN	O	O
in	NN	O	O
downmodulation	NN	O	O
of	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
and	NN	O	I-protein
beta	NN	O	I-protein
isozymes	NN	O	I-protein
of	NN	O	O
PKC	NN	O	B-protein
,	NN	O	O
and	NN	O	O
abrogates	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
treatment	NN	O	O
also	NN	O	O
abolishes	NN	O	O
induction	NN	O	O
of	NN	O	O
chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
Ar-5	NN	O	B-cell_line
cells	NN	O	I-cell_line
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
plasmid	NN	O	B-DNA
containing	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
this	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
including	NN	O	O
NFAT-1	NN	O	B-protein
,	NN	O	O
Oct	NN	O	B-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
and	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
are	NN	O	O
all	NN	O	O
potentially	NN	O	O
sensitive	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
trimer	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT	NN	O	B-DNA
and	NN	O	I-DNA
Oct	NN	O	I-DNA
sites	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
sensitive	NN	O	O
to	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
treatment	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
have	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
inducibility	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
located	NN	O	O
at	NN	O	O
-150	NN	O	B-DNA
bp	NN	O	I-DNA
almost	NN	O	O
completely	NN	O	O
abrogate	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
appearance	NN	O	O
of	NN	O	O
an	NN	O	O
inducible	NN	O	B-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
is	NN	O	O
sensitive	NN	O	O
to	NN	O	O
PKC	NN	O	B-protein
depletion	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
cotransfections	NN	O	O
with	NN	O	O
c-fos	NN	O	O
and	NN	O	O
c-jun	NN	O	O
expression	NN	O	O
plasmids	NN	O	O
markedly	NN	O	O
enhance	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
minimally	NN	O	B-cell_line
stimulated	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
at	NN	O	O
-150	NN	O	B-DNA
bp	NN	O	I-DNA
represents	NN	O	O
a	NN	O	O
major	NN	O	O
,	NN	O	O
if	NN	O	O
not	NN	O	O
the	NN	O	O
only	NN	O	O
,	NN	O	O
site	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
responsiveness	NN	O	O
in	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Effect	NN	O	O
of	NN	O	O
antihypertensive	NN	O	O
therapy	NN	O	O
with	NN	O	O
captopril	NN	O	O
on	NN	O	O
gluco-	NN	O	B-protein
and	NN	O	I-protein
mineralocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
hypertensive	NN	O	O
patients	NN	O	O
of	NN	O	O
various	NN	O	O
age	NN	O	O
]	NN	O	O

Binding	NN	O	O
of	NN	O	O
3H-dexamethasone	NN	O	O
and	NN	O	O
3H-aldosterone	NN	O	O
by	NN	O	O
peripheral	NN	O	B-protein
lymphocyte	NN	O	I-protein
receptors	NN	O	I-protein
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
healthy	NN	O	O
persons	NN	O	O
and	NN	O	O
hypertensive	NN	O	O
patients	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
2-week	NN	O	O
captopril	NN	O	O
treatment	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
and	NN	O	I-protein
mineralocorticoid	NN	O	I-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
was	NN	O	O
increased	NN	O	O
in	NN	O	O
hypertensives	NN	O	O
vs	NN	O	O
normotensives	NN	O	O
.	NN	O	O

The	NN	O	O
treatment	NN	O	O
with	NN	O	O
the	NN	O	O
ACE	NN	O	O
inhibitor	NN	O	O
captopril	NN	O	O
led	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
hormone-receptor	NN	O	O
interactions	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
more	NN	O	O
marked	NN	O	O
rise	NN	O	O
of	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
receptors	NN	O	B-protein
in	NN	O	O
middle-aged	NN	O	O
(	NN	O	O
44-55	NN	O	O
years	NN	O	O
)	NN	O	O
hypertensives	NN	O	O
vs	NN	O	O
elderly	NN	O	O
(	NN	O	O
61-80	NN	O	O
years	NN	O	O
)	NN	O	O
subjects	NN	O	O
after	NN	O	O
captopril	NN	O	O
treatment	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
use	NN	O	O
of	NN	O	O
interferon-gamma-treated	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
chemiluminescence	NN	O	O
assays	NN	O	O
to	NN	O	O
detect	NN	O	O
red	NN	O	B-protein
cell	NN	O	I-protein
,	NN	O	I-protein
platelet	NN	O	I-protein
and	NN	O	I-protein
granulocyte	NN	O	I-protein
antibodies	NN	O	I-protein
of	NN	O	O
potential	NN	O	O
clinical	NN	O	O
significance	NN	O	O
.	NN	O	O

The	NN	O	O
chemiluminescent	NN	O	O
(	NN	O	O
CL	NN	O	O
)	NN	O	O
response	NN	O	O
of	NN	O	O
interferon-gamma-treated	NN	O	B-cell_line
U937	NN	O	I-cell_line
(	NN	O	I-cell_line
IFN-U937	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
sensitized	NN	O	O
target	NN	O	O
cells	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
detect	NN	O	O
red	NN	O	B-protein
cell	NN	O	I-protein
,	NN	O	O
platelet	NN	O	B-protein
and	NN	O	O
granulocyte	NN	O	B-protein
antibodies	NN	O	I-protein
.	NN	O	O

A	NN	O	O
clone	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
selected	NN	O	O
which	NN	O	O
expressed	NN	O	O
Fc	NN	O	B-protein
receptor	NN	O	I-protein
I	NN	O	I-protein
(	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RI	NN	O	I-protein
)	NN	O	O
and	NN	O	O
which	NN	O	O
,	NN	O	O
after	NN	O	O
incubation	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
for	NN	O	O
72	NN	O	O
h	NN	O	O
,	NN	O	O
was	NN	O	O
capable	NN	O	O
of	NN	O	O
generating	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
lucigenin-enhanced	NN	O	O
CL	NN	O	O
.	NN	O	O

The	NN	O	O
CL	NN	O	O
responses	NN	O	O
of	NN	O	O
IFN-U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
to	NN	O	O
sensitized	NN	O	B-cell_type
red	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
platelets	NN	O	B-cell_type
or	NN	O	O
granulocytes	NN	O	B-cell_type
were	NN	O	O
then	NN	O	O
compared	NN	O	O
.	NN	O	O

Assays	NN	O	O
using	NN	O	O
monocytes	NN	O	B-cell_type
or	NN	O	O
IFN-U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
of	NN	O	O
comparable	NN	O	O
sensitivity	NN	O	O
for	NN	O	O
detection	NN	O	O
of	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
all	NN	O	O
three	NN	O	O
types	NN	O	O
of	NN	O	O
target	NN	O	O
cell	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
IFN-U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
reduced	NN	O	O
interassay	NN	O	O
variation	NN	O	O
and	NN	O	O
simplified	NN	O	O
assay	NN	O	O
performance	NN	O	O
.	NN	O	O

The	NN	O	O
potential	NN	O	O
clinical	NN	O	O
usefulness	NN	O	O
of	NN	O	O
these	NN	O	O
CL	NN	O	O
assays	NN	O	O
was	NN	O	O
suggested	NN	O	O
by	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
both	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
IFN-U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
respond	NN	O	O
to	NN	O	O
red	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
platelets	NN	O	B-cell_type
or	NN	O	O
granulocytes	NN	O	B-cell_type
sensitized	NN	O	O
with	NN	O	O
sera	NN	O	O
from	NN	O	O
pregnant	NN	O	O
women	NN	O	O
whose	NN	O	O
babies	NN	O	O
had	NN	O	O
either	NN	O	O
haemolytic	NN	O	O
disease	NN	O	O
of	NN	O	O
the	NN	O	O
newborn	NN	O	O
(	NN	O	O
HDN	NN	O	O
)	NN	O	O
,	NN	O	O
alloimmune	NN	O	O
thrombocytopenia	NN	O	O
or	NN	O	O
alloimmune	NN	O	O
neutropenia	NN	O	O
respectively	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
IFN-U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
both	NN	O	O
responded	NN	O	O
to	NN	O	O
red	NN	O	B-cell_type
cells	NN	O	I-cell_type
sensitized	NN	O	O
with	NN	O	O
antibodies	NN	O	B-protein
against	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
specificities	NN	O	O
of	NN	O	O
assumed	NN	O	O
(	NN	O	O
although	NN	O	O
not	NN	O	O
documented	NN	O	O
)	NN	O	O
clinical	NN	O	O
significance	NN	O	O
for	NN	O	O
blood	NN	O	O
transfusion	NN	O	O
recipients	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
IFN-U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
responded	NN	O	O
only	NN	O	O
weakly	NN	O	O
to	NN	O	O
red	NN	O	B-cell_type
cells	NN	O	I-cell_type
sensitized	NN	O	O
with	NN	O	O
either	NN	O	O
anti-D	NN	O	B-protein
in	NN	O	O
sera	NN	O	O
from	NN	O	O
mothers	NN	O	O
of	NN	O	O
babies	NN	O	O
unaffected	NN	O	O
by	NN	O	O
HDN	NN	O	O
,	NN	O	O
or	NN	O	O
with	NN	O	O
antisera	NN	O	O
containing	NN	O	O
high	NN	O	B-protein
titre	NN	O	I-protein
antibodies	NN	O	I-protein
with	NN	O	O
specificities	NN	O	O
not	NN	O	O
normally	NN	O	O
associated	NN	O	O
with	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
red	NN	O	B-cell_type
cell	NN	O	I-cell_type
survival	NN	O	O
.	NN	O	O

-DOCSTART-	O

Ablation	NN	O	O
of	NN	O	O
transplanted	NN	O	O
HTLV-I	NN	O	O
Tax-transformed	NN	O	O
tumors	NN	O	O
in	NN	O	O
mice	NN	O	O
by	NN	O	O
antisense	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
Science	NN	O	O
1993	NN	O	O
Mar	NN	O	O
12	NN	O	O
;	NN	O	O
259	NN	O	O
(	NN	O	O
5101	NN	O	O
)	NN	O	O
:	NN	O	O
1523	NN	O	O
]	NN	O	O

Mice	NN	O	O
transgenic	NN	O	O
for	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
leukemia	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
HTLV-I	NN	O	I-DNA
)	NN	O	I-DNA
Tax	NN	O	I-DNA
gene	NN	O	I-DNA
develop	NN	O	O
fibroblastic	NN	O	O
tumors	NN	O	O
that	NN	O	O
express	NN	O	O
NF-kappa	NN	O	B-DNA
B-inducible	NN	O	I-DNA
early	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
antisense	NN	O	O
oligodeoxynucleotides	NN	O	O
(	NN	O	O
ODNs	NN	O	O
)	NN	O	O
inhibited	NN	O	O
growth	NN	O	O
of	NN	O	O
these	NN	O	O
culture-adapted	NN	O	B-cell_line
Tax-transformed	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
as	NN	O	O
an	NN	O	O
HTLV-I-transformed	NN	O	B-cell_line
human	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
antisense	NN	O	O
inhibition	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
itself	NN	O	O
had	NN	O	O
no	NN	O	O
apparent	NN	O	O
effect	NN	O	O
on	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

Mice	NN	O	O
treated	NN	O	O
with	NN	O	O
antisense	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
ODNs	NN	O	O
showed	NN	O	O
rapid	NN	O	O
regression	NN	O	O
of	NN	O	O
transplanted	NN	O	O
fibrosarcomas	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
expression	NN	O	O
may	NN	O	O
be	NN	O	O
necessary	NN	O	O
for	NN	O	O
the	NN	O	O
maintenance	NN	O	O
of	NN	O	O
the	NN	O	O
malignant	NN	O	O
phenotype	NN	O	O
and	NN	O	O
provides	NN	O	O
a	NN	O	O
therapeutic	NN	O	O
approach	NN	O	O
for	NN	O	O
HTLV-I-associated	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Membrane	NN	O	B-protein
receptors	NN	O	I-protein
for	NN	O	O
aldosterone	NN	O	O
:	NN	O	O
a	NN	O	O
novel	NN	O	O
pathway	NN	O	O
for	NN	O	O
mineralocorticoid	NN	O	O
action	NN	O	O
.	NN	O	O

Rapid	NN	O	O
nongenomic	NN	O	O
in	NN	O	O
vitro	NN	O	O
effects	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
on	NN	O	O
intracellular	NN	O	O
electrolytes	NN	O	O
,	NN	O	O
cell	NN	O	O
volume	NN	O	O
,	NN	O	O
and	NN	O	O
Na	NN	O	B-protein
(	NN	O	I-protein
+	NN	O	I-protein
)	NN	O	I-protein
-H+	NN	O	I-protein
antiport	NN	O	I-protein
have	NN	O	O
been	NN	O	O
found	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
HML	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
125I-labeled	NN	O	O
aldosterone	NN	O	O
to	NN	O	O
plasma	NN	O	O
membranes	NN	O	O
of	NN	O	O
HML	NN	O	B-cell_type
shares	NN	O	O
important	NN	O	O
features	NN	O	O
with	NN	O	O
these	NN	O	O
functional	NN	O	O
data	NN	O	O
.	NN	O	O

This	NN	O	O
includes	NN	O	O
a	NN	O	O
very	NN	O	O
low	NN	O	O
apparent	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
of	NN	O	O
0.1	NN	O	O
nM	NN	O	O
for	NN	O	O
both	NN	O	O
aldosterone	NN	O	O
and	NN	O	O
the	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
Na	NN	O	B-protein
(	NN	O	I-protein
+	NN	O	I-protein
)	NN	O	I-protein
-H	NN	O	I-protein
(	NN	O	I-protein
+	NN	O	I-protein
)	NN	O	I-protein
-antiport	NN	O	I-protein
,	NN	O	O
a	NN	O	O
high	NN	O	O
turnover	NN	O	O
rate	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
almost	NN	O	O
exclusive	NN	O	O
binding	NN	O	O
selectivity	NN	O	O
for	NN	O	O
aldosterone	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
,	NN	O	O
RU	NN	O	O
26988	NN	O	O
,	NN	O	O
corticosterone	NN	O	O
,	NN	O	O
ouabain	NN	O	O
,	NN	O	O
amiloride	NN	O	O
,	NN	O	O
and	NN	O	O
18-hydroxyprogesterone	NN	O	O
were	NN	O	O
inactive	NN	O	O
as	NN	O	O
ligands	NN	O	O
.	NN	O	O

Deoxycorticosterone	NN	O	O
acetate	NN	O	O
had	NN	O	O
an	NN	O	O
intermediate	NN	O	O
activity	NN	O	O
with	NN	O	O
an	NN	O	O
apparent	NN	O	O
Kd	NN	O	O
of	NN	O	O
100	NN	O	O
nM	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
are	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
demonstrate	NN	O	O
membrane	NN	O	O
binding	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
being	NN	O	O
compatible	NN	O	O
with	NN	O	O
major	NN	O	O
aspects	NN	O	O
of	NN	O	O
its	NN	O	O
nongenomic	NN	O	O
effects	NN	O	O
.	NN	O	O

-DOCSTART-	O

Leukotriene	NN	O	O
B4	NN	O	O
transcriptionally	NN	O	O
activates	NN	O	O
interleukin-6	NN	O	B-protein
expression	NN	O	O
involving	NN	O	O
NK-chi	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
NF-IL6	NN	O	B-protein
.	NN	O	O

Leukotriene	NN	O	O
B4	NN	O	O
(	NN	O	O
LTB4	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
notable	NN	O	O
participant	NN	O	O
in	NN	O	O
inflammation	NN	O	O
and	NN	O	O
chemotaxis	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
still	NN	O	O
unclear	NN	O	O
whether	NN	O	O
LTB4	NN	O	O
acts	NN	O	O
in	NN	O	O
this	NN	O	O
regard	NN	O	O
directly	NN	O	O
or	NN	O	O
indirectly	NN	O	O
by	NN	O	O
stimulating	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
chemotactic	NN	O	O
and	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
LTB4	NN	O	O
induces	NN	O	O
synthesis	NN	O	O
of	NN	O	O
interleukin	NN	O	B-protein
(	NN	O	I-protein
IL	NN	O	I-protein
)	NN	O	I-protein
-6	NN	O	I-protein
by	NN	O	O
human	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
through	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
furthermore	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
this	NN	O	O
process	NN	O	O
involves	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-chi	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
,	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
,	NN	O	O
of	NN	O	O
NF-IL6	NN	O	B-protein
,	NN	O	O
while	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
,	NN	O	O
shown	NN	O	O
to	NN	O	O
otherwise	NN	O	O
confer	NN	O	O
IL-6	NN	O	B-protein
inducibility	NN	O	O
,	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
unaffected	NN	O	O
by	NN	O	O
LTB4	NN	O	O
.	NN	O	O

Involvement	NN	O	O
of	NN	O	O
NF-chi	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
NF-IL6	NN	O	B-protein
in	NN	O	O
induction	NN	O	O
of	NN	O	O
IL-6	NN	O	B-protein
transcription	NN	O	O
by	NN	O	O
monocytes	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
using	NN	O	O
deleted	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
LTB4	NN	O	O
was	NN	O	O
not	NN	O	O
only	NN	O	O
associated	NN	O	O
with	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
respective	NN	O	O
transcripts	NN	O	O
but	NN	O	O
resulted	NN	O	O
in	NN	O	O
synthesis	NN	O	O
of	NN	O	O
functional	NN	O	O
IL-6	NN	O	B-protein
protein	NN	O	I-protein
as	NN	O	O
well	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
LTB4	NN	O	O
mediated	NN	O	O
transactivation	NN	O	O
of	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
construct	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
NF-chi	NN	O	B-DNA
B	NN	O	I-DNA
or	NN	O	O
the	NN	O	O
NF-IL6	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
signaling	NN	O	O
events	NN	O	O
mediating	NN	O	O
this	NN	O	O
effect	NN	O	O
appeared	NN	O	O
to	NN	O	O
involve	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
H2O2	NN	O	O
,	NN	O	O
since	NN	O	O
LTB4	NN	O	O
failed	NN	O	O
to	NN	O	O
induce	NN	O	O
NF-chi	NN	O	B-protein
B	NN	O	I-protein
or	NN	O	O
NF-IL6	NN	O	B-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
scavenger	NN	O	O
of	NN	O	O
H2O2	NN	O	O
,	NN	O	O
N-acetyl-L-cysteine	NN	O	O
.	NN	O	O

-DOCSTART-	O

Bcl-2	NN	O	B-protein
:	NN	O	O
a	NN	O	O
repressor	NN	O	O
of	NN	O	O
lymphocyte	NN	O	B-cell_type
death	NN	O	O
.	NN	O	O

The	NN	O	O
genes	NN	O	O
and	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
are	NN	O	O
currently	NN	O	O
the	NN	O	O
subject	NN	O	O
of	NN	O	O
intense	NN	O	O
study	NN	O	O
.	NN	O	O

The	NN	O	O
bcl-2	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
repressor	NN	O	O
of	NN	O	O
lymphocyte	NN	O	B-cell_type
death	NN	O	O
,	NN	O	O
is	NN	O	O
perhaps	NN	O	O
the	NN	O	O
best	NN	O	O
understood	NN	O	O
of	NN	O	O
the	NN	O	O
programmed	NN	O	O
cell	NN	O	O
death	NN	O	O
associated	NN	O	O
genes	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
Stanley	NN	O	O
Korsmeyer	NN	O	O
provides	NN	O	O
a	NN	O	O
brief	NN	O	O
overview	NN	O	O
of	NN	O	O
bcl-2	NN	O	B-protein
,	NN	O	O
concentrating	NN	O	O
on	NN	O	O
its	NN	O	O
roles	NN	O	O
in	NN	O	O
B-	NN	O	O
and	NN	O	O
T-cell	NN	O	O
development	NN	O	O
and	NN	O	O
in	NN	O	O
oncogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cytoplasmic	NN	O	O
domain	NN	O	O
heterogeneity	NN	O	O
and	NN	O	O
functions	NN	O	O
of	NN	O	O
IgG	NN	O	B-protein
Fc	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	O
express	NN	O	O
closely	NN	O	O
related	NN	O	O
immunoglobulin	NN	O	B-protein
G	NN	O	I-protein
(	NN	O	I-protein
IgG	NN	O	I-protein
)	NN	O	I-protein
Fc	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RII	NN	O	I-protein
)	NN	O	O
that	NN	O	O
differ	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
structures	NN	O	O
of	NN	O	O
their	NN	O	O
cytoplasmic	NN	O	B-protein
domains	NN	O	I-protein
.	NN	O	O

Because	NN	O	O
of	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
alternative	NN	O	O
messenger	NN	O	O
RNA	NN	O	O
splicing	NN	O	O
,	NN	O	O
B-cell	NN	O	B-protein
Fc	NN	O	I-protein
gamma	NN	O	I-protein
RII	NN	O	I-protein
contains	NN	O	O
an	NN	O	O
insertion	NN	O	O
of	NN	O	O
47	NN	O	O
amino	NN	O	O
acids	NN	O	O
that	NN	O	O
participates	NN	O	O
in	NN	O	O
determining	NN	O	O
receptor	NN	O	O
function	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
an	NN	O	O
Fc	NN	O	B-cell_line
gamma	NN	O	I-cell_line
RII-negative	NN	O	I-cell_line
B-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
with	NN	O	O
complementary	NN	O	O
DNA	NN	O	O
's	NN	O	O
encoding	NN	O	O
the	NN	O	O
two	NN	O	O
splice	NN	O	O
products	NN	O	O
and	NN	O	O
various	NN	O	O
receptor	NN	O	O
mutants	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
insertion	NN	O	O
was	NN	O	O
responsible	NN	O	O
for	NN	O	O
preventing	NN	O	O
both	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RII	NN	O	I-protein
-mediated	NN	O	O
endocytosis	NN	O	O
and	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RII	NN	O	I-protein
-mediated	NN	O	O
antigen	NN	O	O
presentation	NN	O	O
.	NN	O	O

The	NN	O	O
insertion	NN	O	O
was	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RII	NN	O	I-protein
to	NN	O	O
modulate	NN	O	O
surface	NN	O	B-protein
immunoglobulin	NN	O	I-protein
-triggered	NN	O	O
B-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Instead	NN	O	O
,	NN	O	O
regulation	NN	O	O
of	NN	O	O
activation	NN	O	O
involved	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
domain	NN	O	I-protein
common	NN	O	O
to	NN	O	O
both	NN	O	O
the	NN	O	O
lymphocyte	NN	O	B-protein
and	NN	O	I-protein
macrophage	NN	O	I-protein
receptor	NN	O	I-protein
isoforms	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
insertion	NN	O	O
did	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
caps	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
receptor	NN	O	O
cross-linking	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
suggestions	NN	O	O
that	NN	O	O
the	NN	O	O
lymphocyte	NN	O	O
but	NN	O	O
not	NN	O	O
macrophage	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
can	NN	O	O
associate	NN	O	O
with	NN	O	O
the	NN	O	O
detergent-insoluble	NN	O	O
cytoskeleton	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Age-related	NN	O	O
changes	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
and	NN	O	I-protein
mineralocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
healthy	NN	O	O
persons	NN	O	O
and	NN	O	O
patients	NN	O	O
with	NN	O	O
hypertension	NN	O	O
]	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
healthy	NN	O	O
elderly	NN	O	O
subjects	NN	O	O
increases	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
mineralocorticoid	NN	O	O
receptors	NN	O	O
decreases	NN	O	O
.	NN	O	O

The	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
hormone-receptor	NN	O	O
interactions	NN	O	O
in	NN	O	O
hypertension	NN	O	O
are	NN	O	O
activated	NN	O	O
:	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
and	NN	O	I-protein
mineralocorticoid	NN	O	I-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
grows	NN	O	O
in	NN	O	O
hypertensive	NN	O	O
patients	NN	O	O
.	NN	O	O

Still	NN	O	O
a	NN	O	O
more	NN	O	O
essential	NN	O	O
rise	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
receptors	NN	O	O
is	NN	O	O
observed	NN	O	O
in	NN	O	O
mid-age	NN	O	O
hypertensive	NN	O	O
patients	NN	O	O
than	NN	O	O
in	NN	O	O
elderly	NN	O	O
ones	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
.	NN	O	O

CsA	NN	O	O
and	NN	O	O
FK506	NN	O	O
are	NN	O	O
powerful	NN	O	O
suppressors	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
,	NN	O	O
most	NN	O	O
notably	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

They	NN	O	O
act	NN	O	O
at	NN	O	O
a	NN	O	O
point	NN	O	O
in	NN	O	O
activation	NN	O	O
that	NN	O	O
lies	NN	O	O
between	NN	O	O
receptor	NN	O	O
ligation	NN	O	O
and	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
early	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
,	NN	O	O
Stuart	NN	O	O
Schreiber	NN	O	O
and	NN	O	O
Gerald	NN	O	O
Crabtree	NN	O	O
review	NN	O	O
recent	NN	O	O
findings	NN	O	O
that	NN	O	O
indicate	NN	O	O
CsA	NN	O	O
and	NN	O	O
FK506	NN	O	O
operate	NN	O	O
as	NN	O	O
prodrugs	NN	O	O
:	NN	O	O
they	NN	O	O
bind	NN	O	O
endogenous	NN	O	B-protein
intracellular	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
the	NN	O	O
immunophilins	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
resulting	NN	O	O
complex	NN	O	O
targets	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
phosphatase	NN	O	I-protein
,	NN	O	I-protein
calcineurin	NN	O	B-protein
,	NN	O	O
to	NN	O	O
exert	NN	O	O
the	NN	O	O
immunosuppressive	NN	O	O
effect	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-like	NN	O	O
activities	NN	O	O
by	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
1	NN	O	O
infection	NN	O	O
of	NN	O	O
myelomonoblastic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
infection	NN	O	O
on	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
have	NN	O	O
been	NN	O	O
investigated	NN	O	O
in	NN	O	O
a	NN	O	O
new	NN	O	O
model	NN	O	O
of	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

PLB-985	NN	O	B-cell_line
cells	NN	O	I-cell_line
represent	NN	O	O
a	NN	O	O
bipotential	NN	O	B-cell_line
myelomonoblastic	NN	O	I-cell_line
cell	NN	O	I-cell_line
population	NN	O	I-cell_line
capable	NN	O	O
of	NN	O	O
either	NN	O	O
granulocytic	NN	O	O
or	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
after	NN	O	O
induction	NN	O	O
with	NN	O	O
appropriate	NN	O	O
inducers	NN	O	O
.	NN	O	O

By	NN	O	O
virtue	NN	O	O
of	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
on	NN	O	O
the	NN	O	O
cell	NN	O	O
surface	NN	O	O
,	NN	O	O
PLB-985	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
chronically	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
strain	NN	O	O
IIIB	NN	O	O
.	NN	O	O

PLB-IIIB	NN	O	B-cell_line
cells	NN	O	I-cell_line
clearly	NN	O	O
possessed	NN	O	O
a	NN	O	O
more	NN	O	O
monocytic	NN	O	O
phenotype	NN	O	O
than	NN	O	O
the	NN	O	O
parental	NN	O	B-cell_type
myeloblasts	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
differential	NN	O	O
staining	NN	O	O
,	NN	O	O
increased	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
myeloid-specific	NN	O	B-protein
surface	NN	O	I-protein
markers	NN	O	I-protein
,	NN	O	O
and	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
c-fms	NN	O	B-DNA
proto-oncogene	NN	O	I-DNA
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
inducible	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
in	NN	O	O
PLB-985	NN	O	B-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
PLB-IIIB	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
,	NN	O	O
composed	NN	O	O
of	NN	O	O
proteins	NN	O	O
ranging	NN	O	O
between	NN	O	O
70	NN	O	O
and	NN	O	O
110	NN	O	O
kD	NN	O	O
.	NN	O	O

These	NN	O	O
proteins	NN	O	O
interacted	NN	O	O
specifically	NN	O	O
with	NN	O	O
the	NN	O	O
symmetric	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
interferon	NN	O	B-DNA
beta	NN	O	I-DNA
(	NN	O	I-DNA
IFN-beta	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Mutations	NN	O	O
affecting	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
guanine	NN	O	I-DNA
residues	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
were	NN	O	O
unable	NN	O	O
to	NN	O	O
compete	NN	O	O
for	NN	O	O
these	NN	O	O
NF-kappa	NN	O	B-protein
B-related	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Inducibility	NN	O	O
of	NN	O	O
endogenous	NN	O	O
IFN-beta	NN	O	B-RNA
and	NN	O	I-RNA
IFN-alpha	NN	O	I-RNA
RNA	NN	O	I-RNA
was	NN	O	O
also	NN	O	O
increased	NN	O	O
in	NN	O	O
PLB-IIIB	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
of	NN	O	O
myelomonoblastic	NN	O	B-cell_type
cells	NN	O	I-cell_type
may	NN	O	O
select	NN	O	O
for	NN	O	O
a	NN	O	O
more	NN	O	O
mature	NN	O	O
monocytic	NN	O	O
phenotype	NN	O	O
and	NN	O	O
that	NN	O	O
unique	NN	O	O
subunit	NN	O	O
associations	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
differential	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-mediated	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cortisol	NN	O	O
receptor	NN	O	O
resistance	NN	O	O
:	NN	O	O
the	NN	O	O
variability	NN	O	O
of	NN	O	O
its	NN	O	O
clinical	NN	O	O
presentation	NN	O	O
and	NN	O	O
response	NN	O	O
to	NN	O	O
treatment	NN	O	O
.	NN	O	O

Primary	NN	O	O
(	NN	O	O
partial	NN	O	O
)	NN	O	O
cortisol	NN	O	B-protein
receptor	NN	O	I-protein
resistance	NN	O	O
was	NN	O	O
previously	NN	O	O
reported	NN	O	O
in	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
7	NN	O	O
patients	NN	O	O
and	NN	O	O
14	NN	O	O
asymptomatic	NN	O	O
family	NN	O	O
members	NN	O	O
.	NN	O	O

Its	NN	O	O
occurrence	NN	O	O
is	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
extremely	NN	O	O
rare	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
study	NN	O	O
we	NN	O	O
report	NN	O	O
on	NN	O	O
6	NN	O	O
patients	NN	O	O
(	NN	O	O
2	NN	O	O
males	NN	O	O
and	NN	O	O
4	NN	O	O
females	NN	O	O
)	NN	O	O
with	NN	O	O
the	NN	O	O
syndrome	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
male	NN	O	O
patient	NN	O	O
presented	NN	O	O
with	NN	O	O
mild	NN	O	O
hypertension	NN	O	O
.	NN	O	O

Hydrochlorothiazide	NN	O	O
therapy	NN	O	O
resulted	NN	O	O
in	NN	O	O
life-threatening	NN	O	O
hypokalemia	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
male	NN	O	O
patient	NN	O	O
had	NN	O	O
slight	NN	O	O
hypertension	NN	O	O
without	NN	O	O
hypokalemia	NN	O	O
.	NN	O	O

All	NN	O	O
four	NN	O	O
female	NN	O	O
patients	NN	O	O
presented	NN	O	O
between	NN	O	O
the	NN	O	O
age	NN	O	O
of	NN	O	O
20-30	NN	O	O
yr	NN	O	O
with	NN	O	O
acne	NN	O	O
,	NN	O	O
hirsutism	NN	O	O
,	NN	O	O
and	NN	O	O
irregular	NN	O	O
menstruations	NN	O	O
.	NN	O	O

Low	NN	O	O
dose	NN	O	O
dexamethasone	NN	O	O
therapy	NN	O	O
(	NN	O	O
1-1.5	NN	O	O
mg/day	NN	O	O
)	NN	O	O
was	NN	O	O
of	NN	O	O
clinical	NN	O	O
benefit	NN	O	O
in	NN	O	O
these	NN	O	O
patients	NN	O	O
.	NN	O	O

All	NN	O	O
patients	NN	O	O
showed	NN	O	O
insufficient	NN	O	O
suppression	NN	O	O
of	NN	O	O
serum	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
in	NN	O	O
the	NN	O	O
overnight	NN	O	O
1-mg	NN	O	O
dexamethasone	NN	O	O
test	NN	O	O
.	NN	O	O

The	NN	O	O
diurnal	NN	O	O
rhythm	NN	O	O
of	NN	O	O
ACTH	NN	O	O
and	NN	O	O
cortisol	NN	O	O
was	NN	O	O
intact	NN	O	O
,	NN	O	O
albeit	NN	O	O
at	NN	O	O
an	NN	O	O
elevated	NN	O	O
level	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
normal	NN	O	O
increase	NN	O	O
in	NN	O	O
ACTH	NN	O	O
,	NN	O	O
cortisol	NN	O	O
,	NN	O	O
and	NN	O	O
GH	NN	O	O
(	NN	O	O
except	NN	O	O
in	NN	O	O
one	NN	O	O
obese	NN	O	O
patient	NN	O	O
)	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
insulin-induced	NN	O	O
hypoglycemia	NN	O	O
,	NN	O	O
while	NN	O	O
cortisol	NN	O	O
production	NN	O	O
was	NN	O	O
elevated	NN	O	O
in	NN	O	O
three	NN	O	O
patients	NN	O	O
.	NN	O	O

Circulating	NN	O	O
adrenal	NN	O	O
androgen	NN	O	O
levels	NN	O	O
were	NN	O	O
increased	NN	O	O
in	NN	O	O
all	NN	O	O
patients	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
were	NN	O	O
investigated	NN	O	O
in	NN	O	O
a	NN	O	O
whole	NN	O	O
cell	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
assay	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
first	NN	O	O
male	NN	O	O
patient	NN	O	O
,	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
receptors	NN	O	O
was	NN	O	O
very	NN	O	O
low	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
affinity	NN	O	O
was	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
controls	NN	O	O
.	NN	O	O

A	NN	O	O
lowered	NN	O	O
affinity	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
one	NN	O	O
female	NN	O	O
patient	NN	O	O
,	NN	O	O
while	NN	O	O
a	NN	O	O
lowered	NN	O	O
number	NN	O	O
of	NN	O	O
receptors	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
three	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
second	NN	O	O
male	NN	O	O
patient	NN	O	O
,	NN	O	O
no	NN	O	O
abnormalities	NN	O	O
were	NN	O	O
found	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
bioassay	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	O
action	NN	O	O
we	NN	O	O
also	NN	O	O
measured	NN	O	O
dexamethasone	NN	O	O
suppressibility	NN	O	O
of	NN	O	O
mitogen-stimulated	NN	O	O
incorporation	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
male	NN	O	O
patient	NN	O	O
with	NN	O	O
normal	NN	O	O
receptor	NN	O	O
status	NN	O	O
,	NN	O	O
dexamethasone	NN	O	O
suppressibility	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
healthy	NN	O	O
controls	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
both	NN	O	O
maximal	NN	O	O
suppression	NN	O	O
and	NN	O	O
IC50	NN	O	O
.	NN	O	O

Partial	NN	O	O
cortisol	NN	O	O
receptor	NN	O	O
resistance	NN	O	O
might	NN	O	O
be	NN	O	O
less	NN	O	O
rare	NN	O	O
than	NN	O	O
previously	NN	O	O
thought	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
six	NN	O	O
patients	NN	O	O
presented	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
different	NN	O	O
forms	NN	O	O
can	NN	O	O
be	NN	O	O
recognized	NN	O	O
.	NN	O	O

Therapy	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
successful	NN	O	O
in	NN	O	O
female	NN	O	O
patients	NN	O	O
with	NN	O	O
acne	NN	O	O
and	NN	O	O
hirsutism	NN	O	O
,	NN	O	O
as	NN	O	O
the	NN	O	O
secondary	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
adrenal	NN	O	O
androgens	NN	O	O
was	NN	O	O
effectively	NN	O	O
controlled	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
development	NN	O	O
of	NN	O	O
functionally	NN	O	O
responsive	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
work	NN	O	O
reviewed	NN	O	O
in	NN	O	O
this	NN	O	O
article	NN	O	O
separates	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
into	NN	O	O
four	NN	O	O
phases	NN	O	O
.	NN	O	O

First	NN	O	O
is	NN	O	O
an	NN	O	O
expansion	NN	O	O
phase	NN	O	O
prior	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
rearrangement	NN	O	O
,	NN	O	O
which	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
correlated	NN	O	O
with	NN	O	O
programming	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
some	NN	O	O
response	NN	O	O
genes	NN	O	O
for	NN	O	O
inducibility	NN	O	O
.	NN	O	O

This	NN	O	O
phase	NN	O	O
can	NN	O	O
occur	NN	O	O
to	NN	O	O
some	NN	O	O
extent	NN	O	O
outside	NN	O	O
of	NN	O	O
the	NN	O	O
thymus	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
profound	NN	O	O
T	NN	O	O
cell	NN	O	O
deficit	NN	O	O
of	NN	O	O
nude	NN	O	O
mice	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
thymus	NN	O	O
is	NN	O	O
by	NN	O	O
far	NN	O	O
the	NN	O	O
most	NN	O	O
potent	NN	O	O
site	NN	O	O
for	NN	O	O
inducing	NN	O	O
the	NN	O	O
expansion	NN	O	O
per	NN	O	O
se	NN	O	O
,	NN	O	O
even	NN	O	O
if	NN	O	O
other	NN	O	O
sites	NN	O	O
can	NN	O	O
induce	NN	O	O
some	NN	O	O
response	NN	O	O
acquisition	NN	O	O
.	NN	O	O

Second	NN	O	O
is	NN	O	O
a	NN	O	O
controlled	NN	O	O
phase	NN	O	O
of	NN	O	O
TCR	NN	O	B-DNA
gene	NN	O	I-DNA
rearrangement	NN	O	O
.	NN	O	O

The	NN	O	O
details	NN	O	O
of	NN	O	O
the	NN	O	O
regulatory	NN	O	O
mechanism	NN	O	O
that	NN	O	O
selects	NN	O	O
particular	NN	O	O
loci	NN	O	B-DNA
for	NN	O	O
rearrangement	NN	O	O
are	NN	O	O
still	NN	O	O
not	NN	O	O
known	NN	O	O
.	NN	O	O

It	NN	O	O
seems	NN	O	O
that	NN	O	O
the	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
gamma	NN	O	I-DNA
loci	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
gamma	NN	O	B-cell_type
delta	NN	O	I-cell_type
lineage	NN	O	I-cell_type
may	NN	O	O
not	NN	O	O
always	NN	O	O
take	NN	O	O
place	NN	O	O
at	NN	O	O
a	NN	O	O
developmental	NN	O	O
stage	NN	O	O
strictly	NN	O	O
equivalent	NN	O	O
to	NN	O	O
the	NN	O	O
rearrangement	NN	O	O
of	NN	O	O
TCR	NN	O	B-DNA
beta	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
alpha	NN	O	B-cell_type
beta	NN	O	I-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
clear	NN	O	O
just	NN	O	O
how	NN	O	O
early	NN	O	O
the	NN	O	O
two	NN	O	O
lineages	NN	O	O
diverge	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
TCR	NN	O	B-cell_type
alpha	NN	O	I-cell_type
beta	NN	O	I-cell_type
lineage	NN	O	I-cell_type
,	NN	O	I-cell_type
however	NN	O	O
,	NN	O	O
the	NN	O	O
final	NN	O	O
gene	NN	O	O
rearrangement	NN	O	O
events	NN	O	O
are	NN	O	O
accompanied	NN	O	O
by	NN	O	O
rapid	NN	O	O
proliferation	NN	O	O
and	NN	O	O
an	NN	O	O
interruption	NN	O	O
in	NN	O	O
cellular	NN	O	O
response	NN	O	O
gene	NN	O	O
inducibility	NN	O	O
.	NN	O	O

The	NN	O	O
loss	NN	O	O
of	NN	O	O
conventional	NN	O	O
responsiveness	NN	O	O
is	NN	O	O
probably	NN	O	O
caused	NN	O	O
by	NN	O	O
alterations	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
signaling	NN	O	O
,	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
manifestation	NN	O	O
of	NN	O	O
the	NN	O	O
physiological	NN	O	O
state	NN	O	O
that	NN	O	O
is	NN	O	O
a	NN	O	O
precondition	NN	O	O
for	NN	O	O
selection	NN	O	O
.	NN	O	O

Third	NN	O	O
is	NN	O	O
the	NN	O	O
complex	NN	O	O
process	NN	O	O
of	NN	O	O
selection	NN	O	O
.	NN	O	O

Whereas	NN	O	O
peripheral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
undergo	NN	O	O
forms	NN	O	O
of	NN	O	O
positive	NN	O	O
selection	NN	O	O
(	NN	O	O
by	NN	O	O
antigen-driven	NN	O	O
clonal	NN	O	O
expansion	NN	O	O
)	NN	O	O
and	NN	O	O
negative	NN	O	O
selection	NN	O	O
(	NN	O	O
by	NN	O	O
abortive	NN	O	O
stimulation	NN	O	O
leading	NN	O	O
to	NN	O	O
anergy	NN	O	O
or	NN	O	O
death	NN	O	O
)	NN	O	O
,	NN	O	O
neither	NN	O	O
is	NN	O	O
exactly	NN	O	O
the	NN	O	O
same	NN	O	O
phenomenon	NN	O	O
that	NN	O	O
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
thymic	NN	O	O
cortex	NN	O	O
.	NN	O	O

Negative	NN	O	O
selection	NN	O	O
in	NN	O	O
the	NN	O	O
cortex	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
suicidal	NN	O	O
inversion	NN	O	O
of	NN	O	O
antigen	NN	O	O
responsiveness	NN	O	O
:	NN	O	O
instead	NN	O	O
of	NN	O	O
turning	NN	O	O
on	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
the	NN	O	O
activated	NN	O	B-cell_type
cell	NN	O	I-cell_type
destroys	NN	O	O
its	NN	O	O
own	NN	O	O
chromatin	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
genes	NN	O	O
that	NN	O	O
need	NN	O	O
to	NN	O	O
be	NN	O	O
induced	NN	O	O
for	NN	O	O
this	NN	O	O
response	NN	O	O
are	NN	O	O
not	NN	O	O
yet	NN	O	O
identified	NN	O	O
,	NN	O	O
but	NN	O	O
it	NN	O	O
is	NN	O	O
unquestionably	NN	O	O
a	NN	O	O
form	NN	O	O
of	NN	O	O
activation	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
interesting	NN	O	O
that	NN	O	O
in	NN	O	O
humans	NN	O	O
and	NN	O	O
rats	NN	O	O
,	NN	O	O
cortical	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
undergoing	NN	O	O
negative	NN	O	O
selection	NN	O	O
can	NN	O	O
still	NN	O	O
induce	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
even	NN	O	O
be	NN	O	O
rescued	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
if	NN	O	O
exogenous	NN	O	B-protein
IL-2	NN	O	I-protein
is	NN	O	O
provided	NN	O	O
.	NN	O	O

Perhaps	NN	O	O
murine	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
are	NN	O	O
denied	NN	O	O
this	NN	O	O
form	NN	O	O
of	NN	O	O
rescue	NN	O	O
because	NN	O	O
they	NN	O	O
shut	NN	O	O
off	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
chain	NN	O	I-protein
expression	NN	O	O
at	NN	O	O
an	NN	O	O
earlier	NN	O	O
stage	NN	O	O
or	NN	O	O
because	NN	O	O
they	NN	O	O
may	NN	O	O
be	NN	O	O
uncommonly	NN	O	O
Bcl-2	NN	O	B-protein
deficient	NN	O	O
(	NN	O	O
cf.	NN	O	O
Sentman	NN	O	O
et	NN	O	O
al.	NN	O	O
,	NN	O	O
1991	NN	O	O
;	NN	O	O
Strasser	NN	O	O
et	NN	O	O
al.	NN	O	O
,	NN	O	O
1991	NN	O	O
)	NN	O	O
.	NN	O	O

Even	NN	O	O
so	NN	O	O
,	NN	O	O
medullary	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
remain	NN	O	O
at	NN	O	O
least	NN	O	O
partially	NN	O	O
susceptible	NN	O	O
to	NN	O	O
negative	NN	O	O
selection	NN	O	O
even	NN	O	O
as	NN	O	O
they	NN	O	O
continue	NN	O	O
to	NN	O	O
mature	NN	O	O
.	NN	O	O

-DOCSTART-	O

I	NN	O	B-protein
kappa	NN	O	I-protein
B/MAD-3	NN	O	I-protein
masks	NN	O	O
the	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
signal	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
and	NN	O	O
requires	NN	O	O
the	NN	O	O
transactivation	NN	O	B-protein
domain	NN	O	I-protein
to	NN	O	O
inhibit	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

The	NN	O	O
active	NN	O	O
nuclear	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complex	NN	O	I-protein
is	NN	O	O
composed	NN	O	O
of	NN	O	O
two	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
subunits	NN	O	I-protein
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	I-protein
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
share	NN	O	O
extensive	NN	O	O
N-terminal	NN	O	O
sequence	NN	O	O
homology	NN	O	O
with	NN	O	O
the	NN	O	O
v-rel	NN	O	B-protein
oncogene	NN	O	I-protein
product	NN	O	I-protein
.	NN	O	O

The	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
subunit	NN	O	I-protein
provides	NN	O	O
the	NN	O	O
transactivation	NN	O	O
activity	NN	O	O
in	NN	O	O
this	NN	O	O
complex	NN	O	O
and	NN	O	O
serves	NN	O	O
as	NN	O	O
an	NN	O	O
intracellular	NN	O	O
receptor	NN	O	O
for	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
termed	NN	O	O
I	NN	O	O
kappa	NN	O	O
B	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	I-protein
alone	NN	O	O
fails	NN	O	O
to	NN	O	O
stimulate	NN	O	O
kappa	NN	O	O
B-directed	NN	O	O
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
based	NN	O	O
on	NN	O	O
prior	NN	O	O
in	NN	O	O
vitro	NN	O	O
studies	NN	O	O
,	NN	O	O
is	NN	O	O
not	NN	O	O
directly	NN	O	O
regulated	NN	O	O
by	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
critical	NN	O	O
regulatory	NN	O	O
interaction	NN	O	O
between	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B/MAD-3	NN	O	I-protein
,	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
human	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
p65	NN	O	I-protein
mutants	NN	O	I-protein
was	NN	O	O
identified	NN	O	O
that	NN	O	O
functionally	NN	O	O
segregated	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
-mediated	NN	O	O
inhibition	NN	O	O
,	NN	O	O
and	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
-induced	NN	O	O
nuclear	NN	O	O
exclusion	NN	O	O
of	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

Results	NN	O	O
from	NN	O	O
in	NN	O	O
vivo	NN	O	O
expression	NN	O	O
studies	NN	O	O
performed	NN	O	O
with	NN	O	O
these	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
mutants	NN	O	I-protein
revealed	NN	O	O
the	NN	O	O
following	NN	O	O
:	NN	O	O
1	NN	O	O
)	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B/MAD-3	NN	O	I-protein
completely	NN	O	O
inhibits	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
-dependent	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B/MAD-3	NN	O	I-protein
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
retarget	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
from	NN	O	O
the	NN	O	O
nucleus	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
,	NN	O	O
3	NN	O	O
)	NN	O	O
selective	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
functional	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
signal	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
Rel	NN	O	B-protein
homology	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
disrupts	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
engage	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B/MAD-3	NN	O	I-protein
,	NN	O	O
and	NN	O	O
4	NN	O	O
)	NN	O	O
the	NN	O	O
unique	NN	O	O
C-terminus	NN	O	B-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
attenuates	NN	O	O
its	NN	O	O
own	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
and	NN	O	O
contains	NN	O	O
sequences	NN	O	O
that	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Together	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
signal	NN	O	O
and	NN	O	O
transactivation	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
constitute	NN	O	O
a	NN	O	O
bipartite	NN	O	O
system	NN	O	O
that	NN	O	O
is	NN	O	O
critically	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
function	NN	O	O
of	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B/MAD-3	NN	O	I-protein
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
our	NN	O	O
in	NN	O	O
vivo	NN	O	O
studies	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B/MAD-3	NN	O	I-protein
binds	NN	O	O
directly	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	I-protein
.	NN	O	O

This	NN	O	O
interaction	NN	O	O
is	NN	O	O
functional	NN	O	O
as	NN	O	O
it	NN	O	O
leads	NN	O	O
to	NN	O	O
retargeting	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50	NN	O	I-protein
from	NN	O	O
the	NN	O	O
nucleus	NN	O	O
to	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
no	NN	O	O
loss	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
observed	NN	O	O
,	NN	O	O
presumably	NN	O	O
reflecting	NN	O	O
the	NN	O	O
unique	NN	O	O
C-terminal	NN	O	B-protein
domain	NN	O	I-protein
that	NN	O	O
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
that	NN	O	O
present	NN	O	O
in	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p65	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
tissue-specific	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
simian	NN	O	B-DNA
virus	NN	O	I-DNA
40	NN	O	I-DNA
enhancer	NN	O	I-DNA
TC-II	NN	O	I-DNA
(	NN	O	I-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
)	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
biochemically	NN	O	O
and	NN	O	O
functionally	NN	O	O
characterized	NN	O	O
a	NN	O	O
new	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NP-TCII	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
nuclei	NN	O	O
from	NN	O	O
unstimulated	NN	O	B-cell_line
T	NN	O	I-cell_line
and	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
but	NN	O	O
is	NN	O	O
not	NN	O	O
found	NN	O	O
in	NN	O	O
nonhematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
factor	NN	O	O
has	NN	O	O
a	NN	O	O
DNA-binding	NN	O	O
specificity	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
but	NN	O	O
is	NN	O	O
unrelated	NN	O	O
to	NN	O	O
this	NN	O	O
or	NN	O	O
other	NN	O	O
Rel	NN	O	B-protein
proteins	NN	O	I-protein
by	NN	O	O
functional	NN	O	O
and	NN	O	O
biochemical	NN	O	O
criteria	NN	O	O
.	NN	O	O

It	NN	O	O
can	NN	O	O
also	NN	O	O
be	NN	O	O
distinguished	NN	O	O
from	NN	O	O
other	NN	O	O
previously	NN	O	O
described	NN	O	O
lymphocyte-specific	NN	O	B-protein
DNA-binding	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
candidate	NN	O	B-protein
oncoprotein	NN	O	I-protein
Bcl-3	NN	O	I-protein
is	NN	O	O
an	NN	O	O
antagonist	NN	O	O
of	NN	O	O
p50/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-mediated	NN	O	O
inhibition	NN	O	O
.	NN	O	O

The	NN	O	O
candidate	NN	O	B-DNA
oncogene	NN	O	I-DNA
bcl-3	NN	O	I-DNA
was	NN	O	O
discovered	NN	O	O
as	NN	O	O
a	NN	O	O
translocation	NN	O	O
into	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
alpha-locus	NN	O	I-DNA
in	NN	O	O
some	NN	O	O
cases	NN	O	O
of	NN	O	O
B-cell	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukaemias	NN	O	O
.	NN	O	O

The	NN	O	O
protein	NN	O	B-protein
Bcl-3	NN	O	I-protein
contains	NN	O	O
seven	NN	O	O
so-called	NN	O	O
ankyrin	NN	O	B-protein
repeats	NN	O	I-protein
.	NN	O	O

Similar	NN	O	O
repeat	NN	O	B-protein
motifs	NN	O	I-protein
are	NN	O	O
found	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
diverse	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
but	NN	O	O
the	NN	O	O
motifs	NN	O	O
of	NN	O	O
Bcl-3	NN	O	B-protein
are	NN	O	O
most	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
those	NN	O	O
found	NN	O	O
in	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
which	NN	O	O
the	NN	O	O
ankyrin	NN	O	B-protein
repeat	NN	O	I-protein
domain	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
directly	NN	O	O
involved	NN	O	O
in	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

No	NN	O	O
biological	NN	O	O
function	NN	O	O
has	NN	O	O
yet	NN	O	O
been	NN	O	O
described	NN	O	O
for	NN	O	O
Bcl-3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
it	NN	O	O
was	NN	O	O
noted	NN	O	O
recently	NN	O	O
that	NN	O	O
Bcl-3	NN	O	B-protein
interferes	NN	O	O
with	NN	O	O
DNA-binding	NN	O	O
of	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
Bcl-3	NN	O	B-protein
can	NN	O	O
aid	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
-dependent	NN	O	O
transcription	NN	O	O
in	NN	O	O
vivo	NN	O	O
by	NN	O	O
counteracting	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
of	NN	O	O
p50/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
homodimers	NN	O	I-protein
.	NN	O	O

Bcl-3	NN	O	B-protein
may	NN	O	O
therefore	NN	O	O
aid	NN	O	O
activation	NN	O	O
of	NN	O	O
select	NN	O	O
NF-kappa	NN	O	B-DNA
B-regulated	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
steroid	NN	O	O
hormones.	NN	O	O
I.	NN	O	O
Estrogens	NN	O	O
]	NN	O	O

The	NN	O	O
steroid	NN	O	O
hormone	NN	O	O
are	NN	O	O
very	NN	O	O
versatile	NN	O	O
molecules	NN	O	O
:	NN	O	O
although	NN	O	O
they	NN	O	O
are	NN	O	O
related	NN	O	O
among	NN	O	O
them	NN	O	O
by	NN	O	O
their	NN	O	O
chemical	NN	O	O
structure	NN	O	O
,	NN	O	O
they	NN	O	O
have	NN	O	O
very	NN	O	O
diverse	NN	O	O
functions	NN	O	O
and	NN	O	O
including	NN	O	O
antagonic	NN	O	O
.	NN	O	O

Their	NN	O	O
action	NN	O	O
mechanism	NN	O	O
is	NN	O	O
not	NN	O	O
completely	NN	O	O
cleared	NN	O	O
.	NN	O	O

The	NN	O	O
estrogens	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
practically	NN	O	O
all	NN	O	O
the	NN	O	O
reproductive	NN	O	O
and	NN	O	O
sexual	NN	O	O
events	NN	O	O
of	NN	O	O
the	NN	O	O
female	NN	O	O
,	NN	O	O
although	NN	O	O
the	NN	O	O
intracellular	NN	O	O
actions	NN	O	O
by	NN	O	O
which	NN	O	O
they	NN	O	O
take	NN	O	O
place	NN	O	O
are	NN	O	O
not	NN	O	O
well	NN	O	O
known	NN	O	O
and	NN	O	O
the	NN	O	O
proposed	NN	O	O
models	NN	O	O
do	NN	O	O
not	NN	O	O
adequately	NN	O	O
satisfy	NN	O	O
the	NN	O	O
questions	NN	O	O
.	NN	O	O

Currently	NN	O	O
it	NN	O	O
is	NN	O	O
accepted	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	B-protein
and/or	NN	O	I-protein
nuclear	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
without	NN	O	O
explaining	NN	O	O
satisfactorily	NN	O	O
how	NN	O	O
the	NN	O	O
hormones	NN	O	O
come	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

The	NN	O	O
endocrine	NN	O	O
events	NN	O	O
that	NN	O	O
are	NN	O	O
rapidly	NN	O	O
expressed	NN	O	O
(	NN	O	O
seconds	NN	O	O
)	NN	O	O
are	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
possible	NN	O	O
interaction	NN	O	O
with	NN	O	O
cellular	NN	O	O
membrane	NN	O	O
.	NN	O	O

The	NN	O	O
purpose	NN	O	O
of	NN	O	O
this	NN	O	O
review	NN	O	O
is	NN	O	O
to	NN	O	O
analyze	NN	O	O
and	NN	O	O
concilliate	NN	O	O
the	NN	O	O
reported	NN	O	O
data	NN	O	O
on	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
estrogens	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reduced	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
of	NN	O	O
ethyl-methanesulfonate-treated	NN	O	B-cell_line
CEM	NN	O	I-cell_line
subclones	NN	O	I-cell_line
correlates	NN	O	O
with	NN	O	O
a	NN	O	O
blockade	NN	O	O
in	NN	O	O
their	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
signaling	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
described	NN	O	O
the	NN	O	O
isolation	NN	O	O
of	NN	O	O
chemically	NN	O	O
induced	NN	O	O
CEM	NN	O	B-cell_line
subclones	NN	O	I-cell_line
that	NN	O	O
express	NN	O	O
CD4	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
bind	NN	O	O
soluble	NN	O	O
gp120	NN	O	B-protein
,	NN	O	O
yet	NN	O	O
show	NN	O	O
a	NN	O	O
markedly	NN	O	O
reduced	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
infection	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

Two	NN	O	O
subclones	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
have	NN	O	O
an	NN	O	O
abnormal	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
activator	NN	O	O
PMA	NN	O	O
.	NN	O	O

PMA	NN	O	O
treatment	NN	O	O
induced	NN	O	O
CD3	NN	O	B-protein
and	NN	O	I-protein
CD25	NN	O	I-protein
(	NN	O	I-protein
IL-2R	NN	O	I-protein
)	NN	O	I-protein
receptors	NN	O	I-protein
on	NN	O	O
the	NN	O	O
parental	NN	O	O
line	NN	O	O
and	NN	O	O
on	NN	O	O
other	NN	O	O
ethyl-methanesulfonate-derived	NN	O	B-cell_line
subclones	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
on	NN	O	O
these	NN	O	O
two	NN	O	O
mutants	NN	O	O
.	NN	O	O

Direct	NN	O	O
assays	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
activity	NN	O	O
were	NN	O	O
conducted	NN	O	O
.	NN	O	O

Total	NN	O	O
cellular	NN	O	O
PKC	NN	O	B-protein
enzymatic	NN	O	O
activity	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
normal	NN	O	O
in	NN	O	O
these	NN	O	O
subclones	NN	O	O
.	NN	O	O

PMA-induced	NN	O	O
CD4	NN	O	B-protein
down-modulation	NN	O	O
occurred	NN	O	O
normally	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
c-raf	NN	O	B-protein
kinase	NN	O	I-protein
was	NN	O	O
normal	NN	O	O
.	NN	O	O

Since	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
contains	NN	O	O
two	NN	O	O
functional	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
kB	NN	O	I-DNA
(	NN	O	I-DNA
NF-kB	NN	O	I-DNA
)	NN	O	I-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
PMA	NN	O	O
to	NN	O	O
induce	NN	O	O
NF-kB	NN	O	B-protein
binding	NN	O	O
activity	NN	O	O
by	NN	O	O
different	NN	O	O
assays	NN	O	O
.	NN	O	O

Chloramphenicol	NN	O	B-protein
acetyl	NN	O	I-protein
transferase	NN	O	I-protein
(	NN	O	O
CAT	NN	O	B-protein
)	NN	O	O
assays	NN	O	O
using	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
(	NN	O	I-DNA
-139	NN	O	I-DNA
)	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat-CAT	NN	O	I-DNA
construct	NN	O	I-DNA
showed	NN	O	O
no	NN	O	O
PMA	NN	O	O
induction	NN	O	O
of	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
these	NN	O	O
subclones	NN	O	O
(	NN	O	O
unlike	NN	O	O
the	NN	O	O
parental	NN	O	O
line	NN	O	O
and	NN	O	O
other	NN	O	O
subclones	NN	O	O
)	NN	O	O
.	NN	O	O

Okadaic	NN	O	O
acid	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
phosphatases	NN	O	B-protein
1	NN	O	I-protein
and	NN	O	I-protein
2A	NN	O	I-protein
,	NN	O	O
did	NN	O	O
not	NN	O	O
overcome	NN	O	O
the	NN	O	O
defect	NN	O	O
in	NN	O	O
these	NN	O	O
subclones	NN	O	O
.	NN	O	O

Gel	NN	O	O
retardation	NN	O	O
assays	NN	O	O
,	NN	O	O
using	NN	O	O
a	NN	O	O
32P-probe	NN	O	O
containing	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
NF-kB	NN	O	B-protein
probe	NN	O	O
and	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
PMA-treated	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
showed	NN	O	O
significantly	NN	O	O
reduced	NN	O	O
induction	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
NF-kB	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
these	NN	O	O
two	NN	O	O
subclones	NN	O	O
compared	NN	O	O
with	NN	O	O
wild	NN	O	B-protein
type	NN	O	I-protein
CEM	NN	O	I-protein
and	NN	O	O
a	NN	O	O
control	NN	O	O
subclone	NN	O	O
.	NN	O	O

Deoxycholate	NN	O	O
treatment	NN	O	O
of	NN	O	O
cytoplasmic	NN	O	O
extracts	NN	O	O
from	NN	O	O
these	NN	O	O
subclones	NN	O	O
released	NN	O	O
much	NN	O	O
reduced	NN	O	O
NF-kB	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
from	NN	O	O
their	NN	O	O
cytoplasmic	NN	O	O
pools	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
reduced	NN	O	O
levels	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
-induced	NN	O	O
nuclear	NN	O	O
NF-kB	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
two	NN	O	O
T	NN	O	O
cell	NN	O	O
subclones	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
their	NN	O	O
normal	NN	O	O
cell	NN	O	O
growth	NN	O	O
,	NN	O	O
but	NN	O	O
correlated	NN	O	O
with	NN	O	O
a	NN	O	O
pronounced	NN	O	O
reduction	NN	O	O
in	NN	O	O
their	NN	O	O
susceptibility	NN	O	O
to	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
contains	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
.	NN	O	O

The	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
NF-AT	NN	O	I-protein
(	NN	O	O
ref.	NN	O	O
1	NN	O	O
)	NN	O	O
is	NN	O	O
induced	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
stimulated	NN	O	O
through	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
receptor/CD3	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
and	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
interleukin-2	NN	O	B-DNA
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
induction	NN	O	O
.	NN	O	O

Although	NN	O	O
NF-AT	NN	O	B-protein
has	NN	O	O
not	NN	O	O
been	NN	O	O
cloned	NN	O	O
or	NN	O	O
purified	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
evidence	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
a	NN	O	O
major	NN	O	O
target	NN	O	O
for	NN	O	O
immunosuppression	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
and	NN	O	O
FK506	NN	O	O
(	NN	O	O
refs	NN	O	O
2-7	NN	O	O
)	NN	O	O
.	NN	O	O

NF-AT	NN	O	B-protein
induction	NN	O	O
may	NN	O	O
require	NN	O	O
two	NN	O	O
activation-dependent	NN	O	O
events	NN	O	O
:	NN	O	O
the	NN	O	O
CsA-sensitive	NN	O	O
translocation	NN	O	O
of	NN	O	O
a	NN	O	O
pre-existing	NN	O	O
component	NN	O	O
and	NN	O	O
the	NN	O	O
CsA-resistant	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
component	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
the	NN	O	O
newly	NN	O	O
synthesized	NN	O	O
nuclear	NN	O	B-protein
component	NN	O	I-protein
of	NN	O	O
NF-AT	NN	O	B-protein
is	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
inducible	NN	O	O
nuclear	NN	O	O
form	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
contains	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
proteins	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
identify	NN	O	O
a	NN	O	O
pre-existing	NN	O	B-protein
NF-AT-binding	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
hypotonic	NN	O	O
extracts	NN	O	O
of	NN	O	O
unstimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

On	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
binding	NN	O	O
,	NN	O	O
reconstitution	NN	O	O
and	NN	O	O
cotransfection	NN	O	O
experiments	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
occurs	NN	O	O
in	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
stages	NN	O	O
:	NN	O	O
a	NN	O	O
CsA-sensitive	NN	O	O
stage	NN	O	O
involving	NN	O	O
modification	NN	O	O
and/or	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
pre-existing	NN	O	O
NF-AT	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
CsA-insensitive	NN	O	O
stage	NN	O	O
involving	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
newly	NN	O	O
synthesized	NN	O	O
Fos	NN	O	B-protein
or	NN	O	O
Fos/Jun	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
the	NN	O	O
pre-existing	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
lymphoid	NN	O	B-protein
cell-specific	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
containing	NN	O	O
c-Rel-like	NN	O	B-protein
proteins	NN	O	I-protein
preferentially	NN	O	O
interacts	NN	O	O
with	NN	O	O
interleukin-6	NN	O	B-DNA
kappa	NN	O	I-DNA
B-related	NN	O	I-DNA
motifs	NN	O	I-DNA
whose	NN	O	O
activities	NN	O	O
are	NN	O	O
repressed	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
proto-oncoprotein	NN	O	B-protein
c-Rel	NN	O	I-protein
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
which	NN	O	O
includes	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
subunits	NN	O	I-protein
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
c-Rel	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
as	NN	O	O
homodimers	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
heterodimers	NN	O	B-protein
with	NN	O	O
p50	NN	O	B-protein
.	NN	O	O

These	NN	O	O
homodimers	NN	O	B-protein
and	NN	O	O
heterodimers	NN	O	B-protein
show	NN	O	O
distinct	NN	O	O
DNA-binding	NN	O	O
specificities	NN	O	O
and	NN	O	O
affinities	NN	O	O
for	NN	O	O
various	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
particular	NN	O	O
,	NN	O	O
the	NN	O	O
c-Rel	NN	O	B-protein
homodimer	NN	O	I-protein
has	NN	O	O
a	NN	O	O
high	NN	O	O
affinity	NN	O	O
for	NN	O	O
interleukin-6	NN	O	B-protein
(	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
and	NN	O	O
beta	NN	O	B-DNA
interferon	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
spite	NN	O	O
of	NN	O	O
its	NN	O	O
association	NN	O	O
with	NN	O	O
p50	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
a	NN	O	O
lymphoid	NN	O	B-protein
cell-specific	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
vivo	NN	O	O
that	NN	O	O
contains	NN	O	O
c-Rel	NN	O	B-protein
but	NN	O	O
not	NN	O	O
p50	NN	O	B-protein
epitopes	NN	O	I-protein
;	NN	O	O
this	NN	O	O
factor	NN	O	O
,	NN	O	O
termed	NN	O	O
IL-6	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
II	NN	O	I-protein
,	NN	O	O
appears	NN	O	O
to	NN	O	O
contain	NN	O	O
the	NN	O	O
c-Rel	NN	O	B-protein
homodimer	NN	O	I-protein
and	NN	O	O
preferentially	NN	O	O
recognizes	NN	O	O
several	NN	O	O
IL-6	NN	O	B-DNA
kappa	NN	O	I-DNA
B-related	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

Although	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
IL-6	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
motif	NN	O	I-DNA
functions	NN	O	O
as	NN	O	O
a	NN	O	O
potent	NN	O	O
IL-1/tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor-responsive	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
nonlymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
its	NN	O	O
activity	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
repressed	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
such	NN	O	O
as	NN	O	O
a	NN	O	O
Jurkat	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
also	NN	O	O
present	NN	O	O
evidence	NN	O	O
that	NN	O	O
IL-6	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
II	NN	O	I-protein
functions	NN	O	O
as	NN	O	O
a	NN	O	O
repressor	NN	O	O
specific	NN	O	O
for	NN	O	O
IL-6	NN	O	B-DNA
kappa	NN	O	I-DNA
B-related	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
motifs	NN	O	I-DNA
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Cortisol	NN	O	O
resistance	NN	O	O
in	NN	O	O
acquired	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
concerns	NN	O	O
9	NN	O	O
iv	NN	O	O
drug	NN	O	O
abusers	NN	O	O
with	NN	O	O
acquired	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
(	NN	O	O
AIDS	NN	O	O
)	NN	O	O
who	NN	O	O
developed	NN	O	O
hypercortisolism	NN	O	O
without	NN	O	O
the	NN	O	O
clinical	NN	O	O
signs	NN	O	O
or	NN	O	O
metabolic	NN	O	O
consequences	NN	O	O
of	NN	O	O
hypercortisolism	NN	O	O
.	NN	O	O

All	NN	O	O
patients	NN	O	O
were	NN	O	O
characterized	NN	O	O
by	NN	O	O
an	NN	O	O
Addisonian	NN	O	O
picture	NN	O	O
(	NN	O	O
weakness	NN	O	O
,	NN	O	O
weight	NN	O	O
loss	NN	O	O
,	NN	O	O
hypotension	NN	O	O
,	NN	O	O
hyponatremia	NN	O	O
,	NN	O	O
and	NN	O	O
intense	NN	O	O
mucocutaneous	NN	O	O
melanosis	NN	O	O
)	NN	O	O
.	NN	O	O

An	NN	O	O
acquired	NN	O	O
form	NN	O	O
of	NN	O	O
peripheral	NN	O	O
resistance	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
was	NN	O	O
suspected	NN	O	O
.	NN	O	O

We	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
examined	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
characteristics	NN	O	O
on	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
by	NN	O	O
measuring	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
and	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
on	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
activation	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
density	NN	O	O
was	NN	O	O
increased	NN	O	O
in	NN	O	O
AIDS	NN	O	O
patients	NN	O	O
with	NN	O	O
an	NN	O	O
Addisonian	NN	O	O
picture	NN	O	O
(	NN	O	O
group	NN	O	O
1	NN	O	O
;	NN	O	O
16.2	NN	O	O
+/-	NN	O	O
9.4	NN	O	O
fmol/million	NN	O	O
cells	NN	O	O
)	NN	O	O
compared	NN	O	O
to	NN	O	O
values	NN	O	O
in	NN	O	O
12	NN	O	O
AIDS	NN	O	O
patients	NN	O	O
without	NN	O	O
an	NN	O	O
Addisonian	NN	O	O
picture	NN	O	O
(	NN	O	O
group	NN	O	O
2	NN	O	O
;	NN	O	O
6.05	NN	O	O
+/-	NN	O	O
2.6	NN	O	O
fmol/million	NN	O	O
cells	NN	O	O
;	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.01	NN	O	O
)	NN	O	O
and	NN	O	O
sex-	NN	O	O
and	NN	O	O
age-matched	NN	O	O
controls	NN	O	O
(	NN	O	O
3.15	NN	O	O
+/-	NN	O	O
2.3	NN	O	O
fmol/million	NN	O	O
cells	NN	O	O
;	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
affinity	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
was	NN	O	O
strikingly	NN	O	O
decreased	NN	O	O
(	NN	O	O
9.36	NN	O	O
+/-	NN	O	O
3.44	NN	O	O
nM	NN	O	O
in	NN	O	O
group	NN	O	O
1	NN	O	O
;	NN	O	O
3.2	NN	O	O
+/-	NN	O	O
1.5	NN	O	O
nM	NN	O	O
in	NN	O	O
group	NN	O	O
2	NN	O	O
;	NN	O	O
2.0	NN	O	O
+/-	NN	O	O
0.8	NN	O	O
nM	NN	O	O
in	NN	O	O
controls	NN	O	O
;	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

[	NN	O	O
3H	NN	O	O
]	NN	O	O
Thymidine	NN	O	O
incorporation	NN	O	O
was	NN	O	O
decreased	NN	O	O
dose-dependently	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
in	NN	O	O
controls	NN	O	O
and	NN	O	O
patients	NN	O	O
;	NN	O	O
the	NN	O	O
effect	NN	O	O
was	NN	O	O
significantly	NN	O	O
blunted	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.05	NN	O	O
)	NN	O	O
in	NN	O	O
group	NN	O	O
1	NN	O	O
patients	NN	O	O
,	NN	O	O
which	NN	O	O
suggests	NN	O	O
that	NN	O	O
activation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
impaired	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
abnormality	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
AIDS	NN	O	O
patients	NN	O	O
with	NN	O	O
hypercortisolism	NN	O	O
and	NN	O	O
clinical	NN	O	O
features	NN	O	O
of	NN	O	O
peripheral	NN	O	O
resistance	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
are	NN	O	O
characterized	NN	O	O
by	NN	O	O
abnormal	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
on	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

Resistance	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
implies	NN	O	O
a	NN	O	O
complex	NN	O	O
change	NN	O	O
in	NN	O	O
immune-endocrine	NN	O	O
function	NN	O	O
,	NN	O	O
which	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
course	NN	O	O
of	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
primer	NN	O	O
extension	NN	O	O
method	NN	O	O
to	NN	O	O
detect	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
CAG	NN	O	O
repeats	NN	O	O
in	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
families	NN	O	O
with	NN	O	O
X-linked	NN	O	O
spinal	NN	O	O
and	NN	O	O
bulbar	NN	O	O
muscular	NN	O	O
atrophy	NN	O	O
.	NN	O	O

X-linked	NN	O	O
spinal	NN	O	O
and	NN	O	O
bulbar	NN	O	O
muscular	NN	O	O
atrophy	NN	O	O
(	NN	O	O
SBMA	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
adult-onset	NN	O	O
form	NN	O	O
of	NN	O	O
motor	NN	O	O
neuron	NN	O	O
disease	NN	O	O
,	NN	O	O
was	NN	O	O
recently	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
caused	NN	O	O
by	NN	O	O
amplification	NN	O	O
of	NN	O	O
the	NN	O	O
CAG	NN	O	O
repeats	NN	O	O
in	NN	O	O
the	NN	O	O
androgen	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
a	NN	O	O
simple	NN	O	O
and	NN	O	O
rapid	NN	O	O
strategy	NN	O	O
to	NN	O	O
detect	NN	O	O
the	NN	O	O
precise	NN	O	O
number	NN	O	O
of	NN	O	O
the	NN	O	O
CAGs	NN	O	O
.	NN	O	O

After	NN	O	O
the	NN	O	O
DNA	NN	O	B-DNA
fragment	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
CAG	NN	O	B-DNA
repeats	NN	O	I-DNA
is	NN	O	O
amplified	NN	O	O
by	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
,	NN	O	O
a	NN	O	O
primer	NN	O	O
extension	NN	O	O
is	NN	O	O
carried	NN	O	O
out	NN	O	O
;	NN	O	O
the	NN	O	O
extension	NN	O	O
of	NN	O	O
the	NN	O	O
end-labelled	NN	O	B-DNA
reverse	NN	O	I-DNA
primer	NN	O	I-DNA
adjacent	NN	O	O
to	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
end	NN	O	I-DNA
of	NN	O	O
CAG	NN	O	B-DNA
repeats	NN	O	I-DNA
stops	NN	O	O
at	NN	O	O
the	NN	O	O
first	NN	O	O
T	NN	O	O
after	NN	O	O
CAG	NN	O	B-DNA
repeats	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
incorporation	NN	O	O
of	NN	O	O
dideoxy	NN	O	O
ATP	NN	O	O
in	NN	O	O
the	NN	O	O
reaction	NN	O	O
mixture	NN	O	O
.	NN	O	O

The	NN	O	O
resultant	NN	O	O
primer	NN	O	B-DNA
products	NN	O	I-DNA
are	NN	O	O
analysed	NN	O	O
by	NN	O	O
denaturing	NN	O	O
polyacrylamide	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
and	NN	O	O
autoradiography	NN	O	O
.	NN	O	O

This	NN	O	O
method	NN	O	O
could	NN	O	O
be	NN	O	O
quite	NN	O	O
useful	NN	O	O
to	NN	O	O
detect	NN	O	O
not	NN	O	O
only	NN	O	O
CAG	NN	O	B-DNA
repeats	NN	O	I-DNA
in	NN	O	O
SBMA	NN	O	O
but	NN	O	O
also	NN	O	O
other	NN	O	O
polymorphic	NN	O	B-DNA
dinucleotide	NN	O	I-DNA
and	NN	O	O
trinucleotide	NN	O	B-DNA
repeats	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Cellular	NN	O	O
immune	NN	O	O
and	NN	O	O
cytokine	NN	O	O
pathways	NN	O	O
resulting	NN	O	O
in	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
relevance	NN	O	O
to	NN	O	O
septic	NN	O	O
shock	NN	O	O
.	NN	O	O

Cells	NN	O	O
of	NN	O	O
monocyte	NN	O	O
lineage	NN	O	O
serve	NN	O	O
as	NN	O	O
effector	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
they	NN	O	O
respond	NN	O	O
to	NN	O	O
LPS	NN	O	O
and	NN	O	O
cytokines	NN	O	O
with	NN	O	O
activation	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
effector	NN	O	I-protein
gene	NN	O	I-protein
products	NN	O	I-protein
similar	NN	O	O
to	NN	O	O
those	NN	O	O
elicited	NN	O	O
by	NN	O	O
the	NN	O	O
antigen	NN	O	O
driven	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
response	NN	O	O
to	NN	O	O
antigen	NN	O	O
proceeds	NN	O	O
at	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
helper	NN	O	I-cell_type
cell	NN	O	I-cell_type
level	NN	O	O
through	NN	O	O
two	NN	O	O
independent	NN	O	O
forms	NN	O	O
of	NN	O	O
cellular	NN	O	O
collaboration	NN	O	O
,	NN	O	O
contact	NN	O	O
and	NN	O	O
lymphokine	NN	O	B-protein
.	NN	O	O

We	NN	O	O
review	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Tissue	NN	O	B-DNA
Factor	NN	O	I-DNA
(	NN	O	I-DNA
TF	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
TF	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
enhanced	NN	O	O
initiation	NN	O	O
of	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
TF	NN	O	O
gene	NN	O	O
appears	NN	O	O
to	NN	O	O
require	NN	O	O
engagement	NN	O	O
of	NN	O	O
a	NN	O	O
56	NN	O	O
bp	NN	O	O
LPS	NN	O	B-DNA
Response	NN	O	I-DNA
Element	NN	O	I-DNA
,	NN	O	O
an	NN	O	O
enhancer	NN	O	O
that	NN	O	O
is	NN	O	O
engaged	NN	O	O
by	NN	O	O
both	NN	O	O
AP-1	NN	O	B-protein
type	NN	O	I-protein
heterodimeric	NN	O	I-protein
complexes	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
like	NN	O	I-protein
heterodimeric	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Dissociation	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
from	NN	O	O
Ig	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
by	NN	O	O
cytokine	NN	O	O
and	NN	O	O
LPS	NN	O	O
stimulation	NN	O	O
,	NN	O	O
and	NN	O	O
possibly	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
may	NN	O	O
represent	NN	O	O
a	NN	O	O
common	NN	O	O
pathway	NN	O	O
to	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
TF	NN	O	B-DNA
and	NN	O	I-DNA
other	NN	O	I-DNA
inflammatory	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Enhancement	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
is	NN	O	O
observed	NN	O	O
upon	NN	O	O
adhesion	NN	O	O
of	NN	O	O
Mo	NN	O	O
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
extracellular	NN	O	O
matrix	NN	O	O
proteins	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
upon	NN	O	O
engagement	NN	O	O
of	NN	O	O
leukocyte	NN	O	B-protein
integrins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
biological	NN	O	O
effects	NN	O	O
that	NN	O	O
follow	NN	O	O
from	NN	O	O
expression	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
by	NN	O	O
vascular	NN	O	O
cells	NN	O	O
have	NN	O	O
been	NN	O	O
resolved	NN	O	O
by	NN	O	O
analysis	NN	O	O
of	NN	O	O
function	NN	O	O
aided	NN	O	O
by	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
recombinant	NN	O	B-protein
full	NN	O	I-protein
length	NN	O	I-protein
TF	NN	O	I-protein
and	NN	O	O
truncated	NN	O	B-protein
surface	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
TF	NN	O	B-protein
.	NN	O	O

The	NN	O	O
rules	NN	O	O
of	NN	O	O
assembly	NN	O	O
of	NN	O	O
the	NN	O	O
cognate	NN	O	O
ligands	NN	O	O
of	NN	O	O
TF	NN	O	B-protein
,	NN	O	O
namely	NN	O	O
the	NN	O	O
zymogen	NN	O	B-protein
plasma	NN	O	I-protein
factors	NN	O	I-protein
VII	NN	O	I-protein
and	NN	O	O
the	NN	O	O
serine	NN	O	B-protein
protease	NN	O	I-protein
factor	NN	O	I-protein
VIIa	NN	O	I-protein
,	NN	O	O
with	NN	O	O
the	NN	O	O
soluble	NN	O	B-protein
surface	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
TF	NN	O	B-protein
in	NN	O	O
free	NN	O	O
solution	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
phospholipid	NN	O	O
surfaces	NN	O	O
and	NN	O	O
cell	NN	O	O
surface	NN	O	O
and	NN	O	O
of	NN	O	O
the	NN	O	O
anchored	NN	O	B-protein
TF	NN	O	I-protein
molecule	NN	O	I-protein
have	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
evident	NN	O	O
that	NN	O	O
assembly	NN	O	O
of	NN	O	O
the	NN	O	O
surface	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
TF	NN	O	B-protein
with	NN	O	O
VIIa	NN	O	B-protein
to	NN	O	O
form	NN	O	O
the	NN	O	O
binary	NN	O	O
TF.VIIa	NN	O	B-protein
complex	NN	O	I-protein
induces	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
Kcat	NN	O	O
of	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
VIIa	NN	O	B-protein
for	NN	O	O
small	NN	O	O
peptidyl	NN	O	O
substrates	NN	O	O
and	NN	O	O
more	NN	O	O
profoundly	NN	O	O
for	NN	O	O
protein	NN	O	B-protein
substrate	NN	O	I-protein
.	NN	O	O

This	NN	O	O
provides	NN	O	O
substantial	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
allosteric	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
catalytic	NN	O	O
cleft	NN	O	O
of	NN	O	O
VIIa	NN	O	B-protein
that	NN	O	O
is	NN	O	O
imparted	NN	O	O
by	NN	O	O
binding	NN	O	O
to	NN	O	O
TF	NN	O	B-protein
,	NN	O	O
its	NN	O	O
cognate	NN	O	O
catalytic	NN	O	O
cofactor	NN	O	B-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
also	NN	O	O
evident	NN	O	O
that	NN	O	O
the	NN	O	O
TF.VIIa	NN	O	B-protein
complex	NN	O	I-protein
is	NN	O	O
proteolytically	NN	O	O
active	NN	O	O
and	NN	O	O
can	NN	O	O
activate	NN	O	O
the	NN	O	O
zymogen	NN	O	B-protein
plasma	NN	O	I-protein
factor	NN	O	I-protein
X	NN	O	I-protein
to	NN	O	O
the	NN	O	O
serine	NN	O	B-protein
protease	NN	O	I-protein
Xa	NN	O	I-protein
in	NN	O	O
free	NN	O	O
solution	NN	O	O
,	NN	O	O
inferring	NN	O	O
that	NN	O	O
extended	NN	O	O
substrate	NN	O	O
recognition	NN	O	O
by	NN	O	O
induced	NN	O	O
structural	NN	O	O
loci	NN	O	O
of	NN	O	O
the	NN	O	O
TF.VIIa	NN	O	B-protein
complex	NN	O	I-protein
are	NN	O	O
created	NN	O	O
from	NN	O	O
either	NN	O	O
or	NN	O	O
both	NN	O	O
proteins	NN	O	O
to	NN	O	O
constitute	NN	O	O
a	NN	O	O
new	NN	O	B-protein
recognition	NN	O	I-protein
structure	NN	O	I-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
also	NN	O	O
evident	NN	O	O
that	NN	O	O
association	NN	O	O
of	NN	O	O
X	NN	O	B-protein
with	NN	O	O
charged	NN	O	O
phospholipid	NN	O	O
surfaces	NN	O	O
enhances	NN	O	O
the	NN	O	O
proteolytic	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
zymogen	NN	O	B-protein
by	NN	O	O
increasing	NN	O	O
recognition	NN	O	O
and	NN	O	O
susceptibility	NN	O	O
of	NN	O	O
the	NN	O	O
sessile	NN	O	O
peptide	NN	O	O
bond	NN	O	O
deduced	NN	O	O
from	NN	O	O
the	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
Km	NN	O	O
and	NN	O	O
increased	NN	O	O
Kcat	NN	O	O
.	NN	O	O

-DOCSTART-	O

Phorbol	NN	O	O
ester	NN	O	O
reduces	NN	O	O
constitutive	NN	O	B-protein
nuclear	NN	O	I-protein
NF	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
inhibits	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
mediator	NN	O	O
of	NN	O	O
specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	O
monocytes	NN	O	O
and	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
genome	NN	O	O
in	NN	O	O
promonocytic	NN	O	O
leukemias	NN	O	O
.	NN	O	O

There	NN	O	O
is	NN	O	O
little	NN	O	O
information	NN	O	O
available	NN	O	O
on	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
to	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
a	NN	O	O
32P-labeled	NN	O	O
oligonucleotide	NN	O	O
derived	NN	O	O
from	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
HIV-1	NN	O	I-DNA
)	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
contains	NN	O	O
a	NN	O	O
tandem	NN	O	O
repeat	NN	O	O
of	NN	O	O
the	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sequence	NN	O	I-DNA
,	NN	O	O
as	NN	O	O
a	NN	O	O
probe	NN	O	O
in	NN	O	O
a	NN	O	O
gel	NN	O	O
retardation	NN	O	O
assay	NN	O	O
to	NN	O	O
study	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
this	NN	O	O
assay	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
detected	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
in	NN	O	O
extracts	NN	O	O
of	NN	O	O
nuclei	NN	O	O
from	NN	O	O
normal	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
with	NN	O	O
12-0-tetradecanoyl	NN	O	O
phorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
for	NN	O	O
4-24	NN	O	O
h	NN	O	O
caused	NN	O	O
the	NN	O	O
complete	NN	O	O
disappearance	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
from	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
monocytes	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
result	NN	O	O
was	NN	O	O
obtained	NN	O	O
with	NN	O	O
the	NN	O	O
mature	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
THP-1	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
constitutive	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
SP1	NN	O	B-protein
was	NN	O	O
unaffected	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
TPA	NN	O	O
.	NN	O	O

The	NN	O	O
disappearance	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
from	NN	O	O
the	NN	O	O
nucleus	NN	O	O
was	NN	O	O
concentration	NN	O	O
dependent	NN	O	O
between	NN	O	O
10	NN	O	O
and	NN	O	O
50	NN	O	O
ng/ml	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
.	NN	O	O

In	NN	O	O
THP-1	NN	O	O
cells	NN	O	O
,	NN	O	O
TPA	NN	O	O
also	NN	O	O
induced	NN	O	O
a	NN	O	O
new	NN	O	O
,	NN	O	O
faster-migrating	NN	O	B-protein
NF	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
species	NN	O	I-protein
not	NN	O	O
induced	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Protein	NN	O	O
kinase	NN	O	O
C	NN	O	O
inhibitor	NN	O	O
staurosporine	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
cyclic	NN	O	O
nucleotide-dependent	NN	O	O
protein	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
HA-1004	NN	O	O
,	NN	O	O
also	NN	O	O
dramatically	NN	O	O
reduced	NN	O	O
constitutive	NN	O	O
levels	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
NF	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
TPA	NN	O	O
addition	NN	O	O
to	NN	O	O
monocytes	NN	O	B-cell_type
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
inhibited	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
reverse	NN	O	B-protein
transcriptase	NN	O	I-protein
assays	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
concentration-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
are	NN	O	O
in	NN	O	O
striking	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
nuclear	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
in	NN	O	O
promonocytic	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
U937	NN	O	I-cell_line
and	NN	O	O
HL-60	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
emphasize	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
studying	NN	O	O
cytokine	NN	O	B-protein
regulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
antiinflammatory	NN	O	O
effects	NN	O	O
of	NN	O	O
corticosteroids	NN	O	O
:	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
regulates	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
expression	NN	O	O
of	NN	O	O
endothelial-leukocyte	NN	O	B-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
1	NN	O	I-protein
.	NN	O	O

Corticosteroids	NN	O	O
are	NN	O	O
the	NN	O	O
preeminent	NN	O	O
antiinflammatory	NN	O	O
agents	NN	O	O
although	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
impart	NN	O	O
their	NN	O	O
efficacy	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
defined	NN	O	O
.	NN	O	O

The	NN	O	O
endothelium	NN	O	O
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
inflammation	NN	O	O
by	NN	O	O
directing	NN	O	O
circulating	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
into	NN	O	O
extravascular	NN	O	O
tissues	NN	O	O
by	NN	O	O
expressing	NN	O	O
adhesive	NN	O	B-protein
molecules	NN	O	I-protein
for	NN	O	O
leukocytes	NN	O	B-cell_type
[	NN	O	O
e.g.	NN	O	O
,	NN	O	O
endothelial-leukocyte	NN	O	B-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
ELAM-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
intercellular	NN	O	B-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
ICAM-1	NN	O	B-protein
)	NN	O	O
]	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
determined	NN	O	O
whether	NN	O	O
corticosteroids	NN	O	O
suppress	NN	O	O
inflammation	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
endothelial	NN	O	O
expression	NN	O	O
of	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
for	NN	O	O
neutrophils	NN	O	B-cell_type
(	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
)	NN	O	O
.	NN	O	O

Preincubation	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
endotoxin	NN	O	O
[	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
,	NN	O	O
1	NN	O	O
microgram/ml	NN	O	O
]	NN	O	O
led	NN	O	O
to	NN	O	O
a	NN	O	O
4-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
subsequent	NN	O	O
adherence	NN	O	O
of	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.0001	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
10	NN	O	O
)	NN	O	O
to	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
an	NN	O	O
increase	NN	O	O
that	NN	O	O
was	NN	O	O
markedly	NN	O	O
attenuated	NN	O	O
when	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
treated	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
IC50	NN	O	O
<	NN	O	O
1	NN	O	O
nM	NN	O	O
,	NN	O	O
P	NN	O	O
<	NN	O	O
0.0001	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
6	NN	O	O
or	NN	O	O
7	NN	O	O
)	NN	O	O
during	NN	O	O
preincubation	NN	O	O
with	NN	O	O
LPS	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
steroid	NN	O	B-protein
receptor	NN	O	I-protein
agonist	NN	O	O
cortisol	NN	O	O
(	NN	O	O
10	NN	O	O
microM	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
its	NN	O	O
inactive	NN	O	O
metabolite	NN	O	O
tetrahydrocortisol	NN	O	O
(	NN	O	O
10	NN	O	O
microM	NN	O	O
)	NN	O	O
,	NN	O	O
diminished	NN	O	O
LPS-induced	NN	O	O
endothelial	NN	O	O
cell	NN	O	O
adhesiveness	NN	O	O
.	NN	O	O

Further	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
mediated	NN	O	O
through	NN	O	O
ligation	NN	O	O
of	NN	O	O
corticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
[	NN	O	O
human	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
hGRs	NN	O	B-protein
)	NN	O	O
]	NN	O	O
was	NN	O	O
provided	NN	O	O
by	NN	O	O
experiments	NN	O	O
utilizing	NN	O	O
the	NN	O	O
steroid	NN	O	O
antagonist	NN	O	O
RU-486	NN	O	O
.	NN	O	O

RU-486	NN	O	O
(	NN	O	O
10	NN	O	O
microM	NN	O	O
)	NN	O	O
,	NN	O	O
which	NN	O	O
prevents	NN	O	O
translocation	NN	O	O
of	NN	O	O
ligated	NN	O	O
hGR	NN	O	B-protein
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
dissociation	NN	O	O
of	NN	O	O
hGR	NN	O	B-protein
from	NN	O	O
heat	NN	O	O
shock	NN	O	O
protein	NN	O	O
90	NN	O	O
,	NN	O	O
completely	NN	O	O
aborted	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
on	NN	O	O
adhesiveness	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
P	NN	O	O
<	NN	O	O
0.0005	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
3	NN	O	O
)	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
LPS	NN	O	O
(	NN	O	O
1	NN	O	O
microgram/ml	NN	O	O
)	NN	O	O
stimulated	NN	O	O
transcription	NN	O	O
of	NN	O	O
ELAM-1	NN	O	B-protein
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
membrane-associated	NN	O	B-protein
ELAM-1	NN	O	I-protein
and	NN	O	O
ICAM-1	NN	O	B-protein
,	NN	O	O
as	NN	O	O
shown	NN	O	O
by	NN	O	O
quantitative	NN	O	O
immunofluorescence	NN	O	O
(	NN	O	O
both	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
9	NN	O	O
)	NN	O	O
.	NN	O	O

Dexamethasone	NN	O	O
markedly	NN	O	O
inhibited	NN	O	O
LPS-stimulated	NN	O	O
accumulation	NN	O	O
of	NN	O	O
mRNA	NN	O	O
for	NN	O	O
ELAM-1	NN	O	B-protein
and	NN	O	O
expression	NN	O	O
of	NN	O	O
ELAM-1	NN	O	B-protein
and	NN	O	O
ICAM-1	NN	O	B-protein
(	NN	O	O
IC50	NN	O	O
<	NN	O	O
10	NN	O	O
nM	NN	O	O
,	NN	O	O
both	NN	O	O
P	NN	O	O
<	NN	O	O
0.001	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
4-9	NN	O	O
)	NN	O	O
;	NN	O	O
inhibition	NN	O	O
of	NN	O	O
expression	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
reversed	NN	O	O
by	NN	O	O
RU-486	NN	O	O
(	NN	O	O
both	NN	O	O
P	NN	O	O
<	NN	O	O
0.005	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
4-6	NN	O	O
)	NN	O	O
.	NN	O	O

As	NN	O	O
in	NN	O	O
the	NN	O	O
adhesion	NN	O	O
studies	NN	O	O
,	NN	O	O
cortisol	NN	O	O
but	NN	O	O
not	NN	O	O
tetrahydrocortisol	NN	O	O
inhibited	NN	O	O
expression	NN	O	O
of	NN	O	O
ELAM-1	NN	O	B-protein
and	NN	O	O
ICAM-1	NN	O	B-protein
(	NN	O	O
both	NN	O	O
P	NN	O	O
<	NN	O	O
0.005	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
3	NN	O	O
or	NN	O	O
4	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
(	NN	O	O
1	NN	O	O
mM	NN	O	O
)	NN	O	O
inhibited	NN	O	O
neither	NN	O	O
adhesion	NN	O	O
nor	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
adhesion	NN	O	B-protein
molecules	NN	O	I-protein
.	NN	O	O

These	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
antagonism	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
of	NN	O	O
endotoxin	NN	O	B-protein
-induced	NN	O	O
inflammation	NN	O	O
is	NN	O	O
a	NN	O	O
specific	NN	O	O
instance	NN	O	O
of	NN	O	O
the	NN	O	O
general	NN	O	O
biological	NN	O	O
principle	NN	O	O
that	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
is	NN	O	O
a	NN	O	O
hormone-dependent	NN	O	O
regulator	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Pax-5	NN	O	B-DNA
encodes	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
BSAP	NN	O	B-protein
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
developing	NN	O	O
CNS	NN	O	O
,	NN	O	O
and	NN	O	O
adult	NN	O	O
testis	NN	O	O
.	NN	O	O

BSAP	NN	O	B-protein
has	NN	O	O
been	NN	O	O
identified	NN	O	O
previously	NN	O	O
as	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
is	NN	O	O
expressed	NN	O	O
at	NN	O	O
early	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
late	NN	O	O
,	NN	O	O
stages	NN	O	O
of	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Biochemical	NN	O	O
purification	NN	O	O
and	NN	O	O
cDNA	NN	O	O
cloning	NN	O	O
has	NN	O	O
now	NN	O	O
revealed	NN	O	O
that	NN	O	O
BSAP	NN	O	B-protein
belongs	NN	O	O
to	NN	O	O
the	NN	O	O
family	NN	O	O
of	NN	O	O
paired	NN	O	B-protein
domain	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

BSAP	NN	O	B-protein
is	NN	O	O
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
Pax-5	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
has	NN	O	O
been	NN	O	O
highly	NN	O	O
conserved	NN	O	O
between	NN	O	O
human	NN	O	O
and	NN	O	O
mouse	NN	O	O
.	NN	O	O

An	NN	O	O
intact	NN	O	O
paired	NN	O	O
domain	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
both	NN	O	O
necessary	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
DNA	NN	O	O
binding	NN	O	O
of	NN	O	O
BSAP	NN	O	B-protein
.	NN	O	O

Binding	NN	O	O
studies	NN	O	O
with	NN	O	O
several	NN	O	O
BSAP	NN	O	B-DNA
recognition	NN	O	I-DNA
sequences	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
sequence	NN	O	O
specificity	NN	O	O
of	NN	O	O
BSAP	NN	O	B-protein
differs	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
distantly	NN	O	O
related	NN	O	O
paired	NN	O	B-protein
domain	NN	O	I-protein
protein	NN	O	I-protein
Pax-1	NN	O	I-protein
.	NN	O	O

During	NN	O	O
embryogenesis	NN	O	O
,	NN	O	O
the	NN	O	O
BSAP	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
transiently	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
mesencephalon	NN	O	O
and	NN	O	O
spinal	NN	O	O
cord	NN	O	O
with	NN	O	O
a	NN	O	O
spatial	NN	O	O
and	NN	O	O
temporal	NN	O	O
expression	NN	O	O
pattern	NN	O	O
that	NN	O	O
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
other	NN	O	O
Pax	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
developing	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
(	NN	O	O
CNS	NN	O	O
)	NN	O	O
.	NN	O	O

Later	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
BSAP	NN	O	B-DNA
gene	NN	O	I-DNA
shifts	NN	O	O
to	NN	O	O
the	NN	O	O
fetal	NN	O	O
liver	NN	O	O
where	NN	O	O
it	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
B	NN	O	O
lymphopoiesis	NN	O	O
.	NN	O	O

BSAP	NN	O	B-protein
expression	NN	O	O
persists	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
also	NN	O	O
seen	NN	O	O
in	NN	O	O
the	NN	O	O
testis	NN	O	O
of	NN	O	O
the	NN	O	O
adult	NN	O	O
mouse	NN	O	O
.	NN	O	O

All	NN	O	O
of	NN	O	O
this	NN	O	O
evidence	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
BSAP	NN	O	B-protein
may	NN	O	O
not	NN	O	O
only	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
neural	NN	O	O
development	NN	O	O
and	NN	O	O
spermatogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cell	NN	O	O
cycle-dependent	NN	O	O
initiation	NN	O	O
and	NN	O	O
lineage-dependent	NN	O	O
abrogation	NN	O	O
of	NN	O	O
GATA-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
pure	NN	O	B-cell_type
differentiating	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
programmed	NN	O	O
activation/repression	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
early	NN	O	O
hematopoietic	NN	O	O
differentiation	NN	O	O
has	NN	O	O
not	NN	O	O
yet	NN	O	O
been	NN	O	O
explored	NN	O	O
.	NN	O	O

The	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
GATA-1	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
normal	NN	O	O
erythroid	NN	O	O
development	NN	O	O
and	NN	O	O
regulates	NN	O	O
erythroid-expressed	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
maturing	NN	O	B-cell_type
erythroblasts	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
analyzed	NN	O	O
GATA-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
early	NN	O	O
human	NN	O	O
adult	NN	O	O
hematopoiesis	NN	O	O
by	NN	O	O
using	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
system	NN	O	O
in	NN	O	O
which	NN	O	O
``	NN	O	O
pure	NN	O	O
''	NN	O	O
early	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
are	NN	O	O
induced	NN	O	O
to	NN	O	O
gradual	NN	O	O
and	NN	O	O
synchronized	NN	O	O
differentiation	NN	O	O
selectively	NN	O	O
along	NN	O	O
the	NN	O	O
erythroid	NN	O	O
or	NN	O	O
granulocyte-macrophage	NN	O	O
pathway	NN	O	O
by	NN	O	O
differential	NN	O	O
treatment	NN	O	O
with	NN	O	O
hematopoietic	NN	O	O
growth	NN	O	O
factors	NN	O	O
.	NN	O	O

The	NN	O	O
GATA-1	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
though	NN	O	O
virtually	NN	O	O
silent	NN	O	O
in	NN	O	O
quiescent	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
activated	NN	O	O
after	NN	O	O
entrance	NN	O	O
into	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
hematopoietic	NN	O	O
growth	NN	O	O
factors	NN	O	O
.	NN	O	O

Subsequently	NN	O	O
,	NN	O	O
increasing	NN	O	O
expression	NN	O	O
along	NN	O	O
the	NN	O	O
erythroid	NN	O	O
pathway	NN	O	O
contrasts	NN	O	O
with	NN	O	O
an	NN	O	O
abrupt	NN	O	O
downregulation	NN	O	O
in	NN	O	O
the	NN	O	O
granulocyte-macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
microenvironment-directed	NN	O	O
,	NN	O	O
two-step	NN	O	O
model	NN	O	O
for	NN	O	O
GATA-1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
differentiating	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
that	NN	O	O
involves	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
cycle-dependent	NN	O	O
initiation	NN	O	O
and	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
lineage-dependent	NN	O	O
maintenance	NN	O	O
or	NN	O	O
suppression	NN	O	O
.	NN	O	O

Hypothetically	NN	O	O
,	NN	O	O
on/off	NN	O	O
switches	NN	O	O
of	NN	O	O
lineage-restricted	NN	O	B-protein
transactivators	NN	O	I-protein
may	NN	O	O
underlie	NN	O	O
the	NN	O	O
binary	NN	O	O
fate	NN	O	O
decisions	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Specific	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
subunits	NN	O	I-protein
act	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
Tat	NN	O	B-protein
to	NN	O	O
stimulate	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
transcription	NN	O	O
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
complex	NN	O	I-protein
which	NN	O	O
functions	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
the	NN	O	O
tat-I	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
to	NN	O	O
stimulate	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
transcription	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
specific	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
contribute	NN	O	O
to	NN	O	O
this	NN	O	O
effect	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
abilities	NN	O	O
of	NN	O	O
different	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
subunits	NN	O	I-protein
to	NN	O	O
act	NN	O	O
with	NN	O	O
Tat-I	NN	O	B-protein
to	NN	O	O
stimulate	NN	O	O
transcription	NN	O	O
of	NN	O	O
HIV	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_type
T-leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
p49	NN	O	B-protein
(	NN	O	I-protein
100	NN	O	I-protein
)	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
subunit	NN	O	I-protein
,	NN	O	O
together	NN	O	O
with	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
can	NN	O	O
act	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
Tat-I	NN	O	B-protein
to	NN	O	O
stimulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
HIV-CAT	NN	O	B-DNA
plasmid	NN	O	I-DNA
.	NN	O	O

Little	NN	O	O
effect	NN	O	O
was	NN	O	O
observed	NN	O	O
with	NN	O	O
50-kDa	NN	O	B-protein
forms	NN	O	I-protein
of	NN	O	O
p105	NN	O	B-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
or	NN	O	O
rel	NN	O	B-protein
,	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
p65	NN	O	B-protein
or	NN	O	O
full-length	NN	O	B-protein
c-rel	NN	O	I-protein
,	NN	O	O
which	NN	O	O
do	NN	O	O
not	NN	O	O
stimulate	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
p49	NN	O	B-protein
(	NN	O	I-protein
100	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
p65	NN	O	B-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
can	NN	O	O
act	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
the	NN	O	O
tat-I	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
to	NN	O	O
stimulate	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
HIV	NN	O	B-RNA
RNA	NN	O	I-RNA
.	NN	O	O

-DOCSTART-	O

Stable	NN	O	O
expression	NN	O	O
of	NN	O	O
HB24	NN	O	B-DNA
,	NN	O	O
a	NN	O	O
diverged	NN	O	B-DNA
human	NN	O	I-DNA
homeobox	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
T	NN	O	O
lymphocytes	NN	O	O
induces	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
growth	NN	O	O
.	NN	O	O

A	NN	O	O
diverged	NN	O	B-DNA
homeobox	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
HB24	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
induced	NN	O	O
following	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
,	NN	O	O
was	NN	O	O
introduced	NN	O	O
into	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
a	NN	O	O
constitutive	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Stable	NN	O	O
transfectants	NN	O	O
of	NN	O	O
HB24	NN	O	B-DNA
were	NN	O	O
established	NN	O	O
that	NN	O	O
expressed	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
HB24	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
possessed	NN	O	O
an	NN	O	O
altered	NN	O	O
phenotype	NN	O	O
suggestive	NN	O	O
of	NN	O	O
activated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
number	NN	O	O
of	NN	O	O
genes	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
induced	NN	O	O
following	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
associated	NN	O	O
with	NN	O	O
cell	NN	O	O
growth	NN	O	O
were	NN	O	O
increased	NN	O	O
in	NN	O	O
the	NN	O	O
transfectants	NN	O	B-cell_line
,	NN	O	O
including	NN	O	O
c-fos	NN	O	B-DNA
,	NN	O	O
c-myc	NN	O	B-DNA
,	NN	O	O
c-myb	NN	O	B-DNA
,	NN	O	O
HLA-DR	NN	O	B-DNA
,	NN	O	O
lck	NN	O	B-DNA
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
interleukin-2	NN	O	B-DNA
and	NN	O	O
interleukin-2	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
expression	NN	O	O
by	NN	O	O
transient	NN	O	O
transfection	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
into	NN	O	O
the	NN	O	O
HB24	NN	O	B-cell_line
transfectants	NN	O	I-cell_line
revealed	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
(	NN	O	O
about	NN	O	O
60	NN	O	O
%	NN	O	O
of	NN	O	O
phytohemagglutinin-	NN	O	B-cell_line
and	NN	O	I-cell_line
phorbol	NN	O	I-cell_line
ester-activated	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
that	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
IL-2R	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
the	NN	O	O
increased	NN	O	O
HB24	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
,	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
HB24	NN	O	I-cell_line
transfectants	NN	O	I-cell_line
proliferated	NN	O	O
more	NN	O	O
rapidly	NN	O	O
than	NN	O	O
control	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
stable	NN	O	O
expression	NN	O	O
of	NN	O	O
HB24	NN	O	B-DNA
confers	NN	O	O
an	NN	O	O
activation	NN	O	O
phenotype	NN	O	O
on	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
implicating	NN	O	O
this	NN	O	O
gene	NN	O	O
as	NN	O	O
an	NN	O	O
important	NN	O	O
transcriptional	NN	O	B-protein
factor	NN	O	I-protein
during	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
and	NN	O	O
growth	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
interaction	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
the	NN	O	O
v-erb	NN	O	B-protein
A	NN	O	I-protein
oncoprotein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
v-erb	NN	O	B-DNA
A	NN	O	I-DNA
oncogene	NN	O	I-DNA
of	NN	O	O
avian	NN	O	O
erythroblastosis	NN	O	O
virus	NN	O	O
is	NN	O	O
a	NN	O	O
mutated	NN	O	O
and	NN	O	O
virally	NN	O	O
transduced	NN	O	O
copy	NN	O	O
of	NN	O	O
a	NN	O	O
host	NN	O	B-DNA
cell	NN	O	I-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
a	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
protein	NN	O	O
expressed	NN	O	O
by	NN	O	O
the	NN	O	O
v-erb	NN	O	B-DNA
A	NN	O	I-DNA
oncogene	NN	O	I-DNA
binds	NN	O	O
to	NN	O	O
DNA	NN	O	O
and	NN	O	O
acts	NN	O	O
as	NN	O	O
a	NN	O	O
dominant	NN	O	O
negative	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
the	NN	O	O
closely	NN	O	O
related	NN	O	O
retinoic	NN	O	B-protein
acid	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
v-erb	NN	O	B-protein
A	NN	O	I-protein
protein	NN	O	I-protein
has	NN	O	O
sustained	NN	O	O
two	NN	O	O
amino	NN	O	O
acid	NN	O	O
alterations	NN	O	O
within	NN	O	O
its	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
relative	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
c-erb	NN	O	B-protein
A	NN	O	I-protein
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
,	NN	O	O
at	NN	O	O
serine	NN	O	O
61	NN	O	O
,	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
v-erb	NN	O	B-protein
A	NN	O	I-protein
function	NN	O	O
in	NN	O	O
the	NN	O	O
neoplastic	NN	O	B-cell_type
cell	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
second	NN	O	O
alteration	NN	O	O
,	NN	O	O
at	NN	O	O
threonine	NN	O	O
78	NN	O	O
,	NN	O	O
also	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
,	NN	O	O
although	NN	O	O
more	NN	O	O
indirect	NN	O	O
,	NN	O	O
role	NN	O	O
:	NN	O	O
alteration	NN	O	O
of	NN	O	O
the	NN	O	O
sequence	NN	O	O
at	NN	O	O
threonine	NN	O	O
78	NN	O	O
such	NN	O	O
that	NN	O	O
it	NN	O	O
resembles	NN	O	O
that	NN	O	O
of	NN	O	O
c-erb	NN	O	B-protein
A	NN	O	I-protein
can	NN	O	O
act	NN	O	O
as	NN	O	O
an	NN	O	O
intragenic	NN	O	O
suppressor	NN	O	O
and	NN	O	O
can	NN	O	O
partially	NN	O	O
restore	NN	O	O
function	NN	O	O
to	NN	O	O
a	NN	O	O
v-erb	NN	O	B-protein
A	NN	O	I-protein
protein	NN	O	I-protein
rendered	NN	O	O
defective	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
mutation	NN	O	O
at	NN	O	O
position	NN	O	B-protein
61	NN	O	I-protein
.	NN	O	O

Threonine	NN	O	O
78	NN	O	O
lies	NN	O	O
within	NN	O	O
the	NN	O	O
D-box	NN	O	B-protein
of	NN	O	O
the	NN	O	O
v-erb	NN	O	B-protein
A	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
a	NN	O	O
region	NN	O	O
thought	NN	O	O
to	NN	O	O
mediate	NN	O	O
receptor-receptor	NN	O	O
dimerizations	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
in	NN	O	O
physical	NN	O	O
proximity	NN	O	O
to	NN	O	O
the	NN	O	O
serine	NN	O	O
at	NN	O	O
position	NN	O	B-protein
61	NN	O	I-protein
.	NN	O	O

It	NN	O	O
therefore	NN	O	O
appears	NN	O	O
that	NN	O	O
an	NN	O	O
indirect	NN	O	O
interaction	NN	O	O
occurs	NN	O	O
between	NN	O	O
these	NN	O	O
two	NN	O	O
sites	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
interaction	NN	O	O
is	NN	O	O
crucial	NN	O	O
for	NN	O	O
v-erb	NN	O	B-protein
A	NN	O	I-protein
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

cis-acting	NN	O	B-DNA
sequences	NN	O	I-DNA
required	NN	O	O
for	NN	O	O
inducible	NN	O	O
interleukin-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
function	NN	O	O
bind	NN	O	O
a	NN	O	O
novel	NN	O	O
Ets-related	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
Elf-1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
recent	NN	O	O
definition	NN	O	O
of	NN	O	O
a	NN	O	O
consensus	NN	O	O
DNA	NN	O	B-DNA
binding	NN	O	I-DNA
sequence	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
has	NN	O	O
allowed	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
potential	NN	O	O
Ets	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
and	NN	O	O
enhancers	NN	O	B-DNA
of	NN	O	O
many	NN	O	O
inducible	NN	O	B-DNA
T-cell	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
studies	NN	O	O
described	NN	O	O
in	NN	O	O
this	NN	O	O
report	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
two	NN	O	O
potential	NN	O	O
Ets	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
EBS1	NN	O	O
and	NN	O	O
EBS2	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
are	NN	O	O
conserved	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
murine	NN	O	I-DNA
interleukin-2	NN	O	I-DNA
enhancers	NN	O	I-DNA
.	NN	O	O

Within	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
these	NN	O	O
two	NN	O	O
sites	NN	O	O
are	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
previously	NN	O	O
defined	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
footprints	NN	O	O
,	NN	O	O
NFAT-1	NN	O	B-DNA
and	NN	O	O
NFIL-2B	NN	O	B-DNA
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
and	NN	O	O
methylation	NN	O	O
interference	NN	O	O
analyses	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
EBS1	NN	O	O
and	NN	O	O
EBS2	NN	O	B-DNA
are	NN	O	O
essential	NN	O	O
for	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT-1	NN	O	B-protein
and	NN	O	I-protein
NFIL-2B	NN	O	I-protein
nuclear	NN	O	I-protein
protein	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
mutagenesis	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
inducible	NN	O	O
interleukin-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
function	NN	O	O
requires	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
either	NN	O	O
EBS1	NN	O	O
or	NN	O	O
EBS2	NN	O	B-DNA
.	NN	O	O

Two	NN	O	O
well-characterized	NN	O	O
Ets	NN	O	B-protein
family	NN	O	I-protein
members	NN	O	I-protein
,	NN	O	O
Ets-1	NN	O	B-protein
and	NN	O	O
Ets-2	NN	O	B-protein
,	NN	O	O
are	NN	O	O
reciprocally	NN	O	O
expressed	NN	O	O
during	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
neither	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
bound	NN	O	O
in	NN	O	O
vitro	NN	O	O
to	NN	O	O
EBS1	NN	O	O
or	NN	O	O
EBS2	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
screened	NN	O	O
a	NN	O	O
T-cell	NN	O	B-DNA
cDNA	NN	O	I-DNA
library	NN	O	O
under	NN	O	O
low-stringency	NN	O	O
conditions	NN	O	O
with	NN	O	O
a	NN	O	O
probe	NN	O	O
from	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
of	NN	O	O
Ets-1	NN	O	B-protein
and	NN	O	O
isolated	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
Ets	NN	O	I-protein
family	NN	O	I-protein
member	NN	O	I-protein
,	NN	O	O
Elf-1	NN	O	B-protein
.	NN	O	O

Elf-1	NN	O	B-protein
contains	NN	O	O
a	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
that	NN	O	O
is	NN	O	O
nearly	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
E74	NN	O	B-protein
,	NN	O	O
the	NN	O	O
ecdysone-inducible	NN	O	B-protein
Drosophila	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
required	NN	O	O
for	NN	O	O
metamorphosis	NN	O	O
(	NN	O	O
hence	NN	O	O
the	NN	O	O
name	NN	O	O
Elf-1	NN	O	B-protein
,	NN	O	O
for	NN	O	O
E74-like	NN	O	B-protein
factor	NN	O	I-protein
1	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Elf-1	NN	O	B-protein
bound	NN	O	O
specifically	NN	O	O
to	NN	O	O
both	NN	O	O
EBS1	NN	O	O
and	NN	O	O
EBS2	NN	O	B-DNA
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

It	NN	O	O
also	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
purine-rich	NN	O	B-DNA
CD3R	NN	O	I-DNA
element	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
2	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
inducible	NN	O	O
virus	NN	O	O
expression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
signalling	NN	O	O
through	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
multiple	NN	O	B-protein
Ets	NN	O	I-protein
family	NN	O	I-protein
members	NN	O	I-protein
with	NN	O	O
apparently	NN	O	O
distinct	NN	O	O
DNA	NN	O	O
binding	NN	O	O
specificities	NN	O	O
regulate	NN	O	O
differential	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
and	NN	O	I-cell_type
activated	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Binding	NN	O	O
of	NN	O	O
erythroid	NN	O	B-protein
and	NN	O	I-protein
non-erythroid	NN	O	I-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
to	NN	O	O
the	NN	O	O
silencer	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
epsilon-globin-encoding	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
clarify	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
developmental	NN	O	O
control	NN	O	O
of	NN	O	O
hemoglobin-encoding	NN	O	B-DNA
genes	NN	O	I-DNA
we	NN	O	O
have	NN	O	O
been	NN	O	O
studying	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	B-DNA
in	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
continuous	NN	O	O
culture	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
reported	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-DNA
control	NN	O	I-DNA
element	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
properties	NN	O	O
of	NN	O	O
a	NN	O	O
silencer	NN	O	B-DNA
extending	NN	O	O
from	NN	O	O
-392	NN	O	B-DNA
to	NN	O	I-DNA
-177	NN	O	I-DNA
bp	NN	O	I-DNA
relative	NN	O	O
to	NN	O	O
the	NN	O	O
cap	NN	O	B-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
epsilon-globin-encoding	NN	O	I-DNA
gene	NN	O	I-DNA
[	NN	O	O
Cao	NN	O	O
et	NN	O	O
al.	NN	O	O
,	NN	O	O
Proc.Natl.Acad.Sci.USA	NN	O	O
86	NN	O	O
(	NN	O	O
1989	NN	O	O
)	NN	O	O
5306-5309	NN	O	O
]	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
showed	NN	O	O
that	NN	O	O
this	NN	O	O
silencer	NN	O	B-DNA
has	NN	O	O
stronger	NN	O	O
inhibitory	NN	O	O
activity	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
K562	NN	O	B-cell_line
human	NN	O	I-cell_line
erythroleukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Using	NN	O	O
deletion	NN	O	B-DNA
mutants	NN	O	I-DNA
and	NN	O	O
cis-cloned	NN	O	O
synthetic	NN	O	O
oligodeoxyribonucleotides	NN	O	O
in	NN	O	O
transient	NN	O	O
expression	NN	O	O
assays	NN	O	O
,	NN	O	O
nucleotide	NN	O	O
sequences	NN	O	O
responsible	NN	O	O
for	NN	O	O
this	NN	O	O
effect	NN	O	O
have	NN	O	O
now	NN	O	O
been	NN	O	O
further	NN	O	O
delimited	NN	O	O
to	NN	O	O
44	NN	O	O
bp	NN	O	O
located	NN	O	O
from	NN	O	O
-294	NN	O	B-DNA
to	NN	O	I-DNA
-251	NN	O	I-DNA
bp	NN	O	I-DNA
.	NN	O	O

Gel	NN	O	O
electrophoresis	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
and	NN	O	O
DNaseI	NN	O	B-protein
footprinting	NN	O	O
assays	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
these	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
sequences	NN	O	O
are	NN	O	O
recognized	NN	O	O
differently	NN	O	O
by	NN	O	O
proteins	NN	O	O
present	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
obtained	NN	O	O
from	NN	O	O
HeLa	NN	O	B-cell_line
and	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Two	NN	O	O
binding	NN	O	O
proteins	NN	O	O
are	NN	O	O
detected	NN	O	O
in	NN	O	O
K562	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
,	NN	O	O
while	NN	O	O
only	NN	O	O
one	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
extracts	NN	O	O
from	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Possible	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
these	NN	O	O
proteins	NN	O	O
may	NN	O	O
regulate	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
epsilon-globin-encoding	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
erythroid	NN	O	B-cell_type
and	NN	O	I-cell_type
non-erythroid	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Calcitriol	NN	O	O
:	NN	O	O
a	NN	O	O
hematolymphopoietrope	NN	O	O
?	NN	O	O
[	NN	O	O
editorial	NN	O	O
]	NN	O	O

A	NN	O	O
MEDLINE	NN	O	O
search	NN	O	O
of	NN	O	O
the	NN	O	O
English-language	NN	O	O
literature	NN	O	O
was	NN	O	O
conducted	NN	O	O
using	NN	O	O
the	NN	O	O
indexing	NN	O	O
terms	NN	O	O
'immunology	NN	O	O
,	NN	O	O
calcitriol	NN	O	O
and	NN	O	O
vitamin	NN	O	O
D	NN	O	O
'	NN	O	O
to	NN	O	O
identify	NN	O	O
studies	NN	O	O
indicating	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
calcitriol	NN	O	O
as	NN	O	O
a	NN	O	O
primary	NN	O	O
immunomodulator	NN	O	O
.	NN	O	O

Sixty-six	NN	O	O
papers	NN	O	O
published	NN	O	O
between	NN	O	O
January	NN	O	O
1956	NN	O	O
and	NN	O	O
June	NN	O	O
1991	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Forty-five	NN	O	O
of	NN	O	O
these	NN	O	O
reports	NN	O	O
are	NN	O	O
cited	NN	O	O
in	NN	O	O
this	NN	O	O
review	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
an	NN	O	O
endocrine	NN	O	O
,	NN	O	O
autocrine	NN	O	O
and/or	NN	O	O
paracrine	NN	O	O
role	NN	O	O
for	NN	O	O
calcitriol	NN	O	O
in	NN	O	O
immune	NN	O	O
regulation	NN	O	O
.	NN	O	O

No	NN	O	O
unifying	NN	O	O
hypothesis	NN	O	O
has	NN	O	O
yet	NN	O	O
emerged	NN	O	O
explaining	NN	O	O
this	NN	O	O
collection	NN	O	O
of	NN	O	O
data	NN	O	O
.	NN	O	O

This	NN	O	O
paper	NN	O	O
provides	NN	O	O
a	NN	O	O
brief	NN	O	O
review	NN	O	O
of	NN	O	O
immune	NN	O	O
properties	NN	O	O
currently	NN	O	O
attributed	NN	O	O
to	NN	O	O
calcitriol	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	O
of	NN	O	O
the	NN	O	O
hypersensitive	NN	O	B-DNA
site	NN	O	I-DNA
HS2	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
genome	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
erythroid-specific	NN	O	B-DNA
hypersensitive	NN	O	I-DNA
site	NN	O	I-DNA
HS2	NN	O	I-DNA
enhancer	NN	O	I-DNA
regulates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
downstream	NN	O	B-DNA
beta-like	NN	O	I-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
10-50	NN	O	B-DNA
kilobases	NN	O	I-DNA
away	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
HS2	NN	O	B-DNA
enhancer	NN	O	I-DNA
function	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
employs	NN	O	O
RNA	NN	O	O
protection	NN	O	O
assays	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
HS2	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
transfected	NN	O	B-DNA
recombinant	NN	O	I-DNA
chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
plasmids	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
erythroid	NN	O	B-cell_line
K562	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
which	NN	O	O
the	NN	O	O
HS2	NN	O	B-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
active	NN	O	O
,	NN	O	O
the	NN	O	O
HS2	NN	O	B-DNA
sequence	NN	O	I-DNA
directs	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
long	NN	O	O
enhancer	NN	O	O
transcripts	NN	O	O
that	NN	O	O
are	NN	O	O
initiated	NN	O	O
apparently	NN	O	O
from	NN	O	O
within	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
and	NN	O	O
elongated	NN	O	O
through	NN	O	O
the	NN	O	O
intervening	NN	O	O
DNA	NN	O	O
into	NN	O	O
the	NN	O	O
cis-linked	NN	O	B-DNA
CAT	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
nonerythroid	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
which	NN	O	O
the	NN	O	O
HS2	NN	O	B-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
inactive	NN	O	O
,	NN	O	O
long	NN	O	B-RNA
enhancer	NN	O	I-RNA
transcripts	NN	O	I-RNA
are	NN	O	O
not	NN	O	O
detectable	NN	O	O
.	NN	O	O

Splitting	NN	O	O
the	NN	O	O
HS2	NN	O	B-DNA
enhancer	NN	O	I-DNA
between	NN	O	O
two	NN	O	O
tandem	NN	O	B-DNA
Ap1	NN	O	I-DNA
sites	NN	O	I-DNA
abolishes	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
long	NN	O	B-RNA
enhancer	NN	O	I-RNA
transcripts	NN	O	I-RNA
and	NN	O	O
results	NN	O	O
in	NN	O	O
loss	NN	O	O
of	NN	O	O
enhancer	NN	O	O
function	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
silencing	NN	O	O
of	NN	O	O
the	NN	O	O
cis-linked	NN	O	O
CAT	NN	O	O
gene	NN	O	O
.	NN	O	O

In	NN	O	O
directing	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
RNA	NN	O	B-RNA
through	NN	O	O
the	NN	O	O
intervening	NN	O	O
DNA	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
gene	NN	O	O
by	NN	O	O
a	NN	O	O
tracking	NN	O	O
and	NN	O	O
transcription	NN	O	O
mechanism	NN	O	O
,	NN	O	O
the	NN	O	O
HS2	NN	O	B-DNA
enhancer	NN	O	I-DNA
may	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
open	NN	O	O
up	NN	O	O
the	NN	O	O
chromatin	NN	O	B-DNA
structure	NN	O	I-DNA
of	NN	O	O
a	NN	O	O
gene	NN	O	B-DNA
domain	NN	O	I-DNA
and	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
deliver	NN	O	O
enhancer	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
to	NN	O	O
the	NN	O	O
promoter	NN	O	O
sequence	NN	O	O
where	NN	O	O
they	NN	O	O
may	NN	O	O
stimulate	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
at	NN	O	O
the	NN	O	O
cap	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
T	NN	O	I-protein
lymphocyte	NN	O	I-protein
protein	NN	O	I-protein
which	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
site	NN	O	O
related	NN	O	O
to	NN	O	O
steroid/thyroid	NN	O	B-DNA
hormone	NN	O	I-DNA
receptor	NN	O	I-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
previously	NN	O	O
identified	NN	O	O
a	NN	O	O
T	NN	O	B-protein
lymphocyte	NN	O	I-protein
protein	NN	O	I-protein
which	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
site	NN	O	O
within	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
and	NN	O	O
exerts	NN	O	O
an	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
on	NN	O	O
virus	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
palindromic	NN	O	B-DNA
site	NN	O	I-DNA
(	NN	O	O
site	NN	O	B-DNA
B	NN	O	I-DNA
)	NN	O	O
recognized	NN	O	O
by	NN	O	O
this	NN	O	O
protein	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
palindromic	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
of	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
steroid/thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
characterize	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
protein	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
as	NN	O	O
a	NN	O	O
100	NN	O	O
kD	NN	O	O
protein	NN	O	O
which	NN	O	O
is	NN	O	O
most	NN	O	O
abundant	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
site	NN	O	B-DNA
B	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
200	NN	O	B-protein
kD	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

This	NN	O	O
protein	NN	O	O
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
other	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
steroid/thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
family	NN	O	I-protein
including	NN	O	O
the	NN	O	O
COUP	NN	O	B-protein
protein	NN	O	I-protein
which	NN	O	O
has	NN	O	O
a	NN	O	O
closely	NN	O	O
related	NN	O	O
DNA	NN	O	O
binding	NN	O	O
specificity	NN	O	O
.	NN	O	O

-DOCSTART-	O

TAR	NN	O	B-DNA
-independent	NN	O	O
transactivation	NN	O	O
by	NN	O	O
Tat	NN	O	B-protein
in	NN	O	O
cells	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
CNS	NN	O	O
:	NN	O	O
a	NN	O	O
novel	NN	O	O
mechanism	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
regulation	NN	O	O
.	NN	O	O

The	NN	O	O
Tat	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
productive	NN	O	O
infection	NN	O	O
and	NN	O	O
is	NN	O	O
a	NN	O	O
potential	NN	O	O
target	NN	O	O
for	NN	O	O
antiviral	NN	O	O
therapy	NN	O	O
.	NN	O	O

Tat	NN	O	B-protein
,	NN	O	O
a	NN	O	O
potent	NN	O	O
activator	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
serves	NN	O	O
to	NN	O	O
greatly	NN	O	O
increase	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
transcription	NN	O	O
directed	NN	O	O
by	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
induction	NN	O	O
,	NN	O	O
which	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
component	NN	O	O
in	NN	O	O
the	NN	O	O
progression	NN	O	O
of	NN	O	O
acquired	NN	O	O
immune	NN	O	O
deficiency	NN	O	O
syndrome	NN	O	O
(	NN	O	O
AIDS	NN	O	O
)	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
increased	NN	O	O
transcriptional	NN	O	O
initiation	NN	O	O
,	NN	O	O
increased	NN	O	O
transcriptional	NN	O	O
elongation	NN	O	O
,	NN	O	O
or	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
these	NN	O	O
processes	NN	O	O
.	NN	O	O

Much	NN	O	O
attention	NN	O	O
has	NN	O	O
been	NN	O	O
focused	NN	O	O
on	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
Tat	NN	O	B-protein
with	NN	O	O
a	NN	O	O
specific	NN	O	O
RNA	NN	O	B-RNA
target	NN	O	I-RNA
termed	NN	O	O
TAR	NN	O	B-RNA
(	NN	O	O
transactivation	NN	O	B-RNA
responsive	NN	O	I-RNA
)	NN	O	O
which	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
leader	NN	O	O
sequence	NN	O	O
of	NN	O	O
all	NN	O	O
HIV-1	NN	O	B-RNA
mRNAs	NN	O	I-RNA
.	NN	O	O

This	NN	O	O
interaction	NN	O	O
is	NN	O	O
believed	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
transactivation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
in	NN	O	O
certain	NN	O	O
CNS-derived	NN	O	B-cell_line
cells	NN	O	I-cell_line
Tat	NN	O	B-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
activating	NN	O	O
HIV-1	NN	O	O
through	NN	O	O
a	NN	O	O
TAR	NN	O	B-DNA
-independent	NN	O	O
pathway	NN	O	O
.	NN	O	O

A	NN	O	O
Tat-responsive	NN	O	B-DNA
element	NN	O	I-DNA
is	NN	O	O
found	NN	O	O
upstream	NN	O	O
within	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
that	NN	O	O
in	NN	O	O
glial-derived	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
allows	NN	O	O
transactivation	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
TAR	NN	O	B-RNA
.	NN	O	O

Deletion	NN	O	O
mapping	NN	O	O
and	NN	O	O
hybrid	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
newly	NN	O	O
identified	NN	O	O
Tat-responsive	NN	O	B-DNA
element	NN	O	I-DNA
corresponds	NN	O	O
to	NN	O	O
a	NN	O	O
sequence	NN	O	O
within	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
previously	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
or	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
domain	NN	O	I-DNA
.	NN	O	O

DNA	NN	O	O
band-shift	NN	O	O
analysis	NN	O	O
reveals	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
glial	NN	O	O
cells	NN	O	O
that	NN	O	O
differs	NN	O	O
from	NN	O	O
that	NN	O	O
present	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Further	NN	O	O
,	NN	O	O
we	NN	O	O
observe	NN	O	O
that	NN	O	O
TAR-deleted	NN	O	B-DNA
mutants	NN	O	I-DNA
of	NN	O	O
HIV-1	NN	O	O
demonstrate	NN	O	O
normal	NN	O	O
late	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
glial	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
syncytia	NN	O	O
formation	NN	O	O
and	NN	O	O
production	NN	O	O
of	NN	O	O
viral	NN	O	B-protein
p24	NN	O	I-protein
antigen	NN	O	I-protein
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-2	NN	O	I-protein
activates	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

The	NN	O	O
HTLV-I	NN	O	B-DNA
LTR	NN	O	I-DNA
contains	NN	O	O
three	NN	O	O
conserved	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
known	NN	O	O
as	NN	O	O
21	NN	O	B-DNA
base	NN	O	I-DNA
pair	NN	O	I-DNA
repeats	NN	O	I-DNA
which	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
stimulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
the	NN	O	O
transactivator	NN	O	B-protein
protein	NN	O	I-protein
tax	NN	O	B-protein
.	NN	O	O

Mutagenesis	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeats	NN	O	I-DNA
can	NN	O	O
be	NN	O	O
subdivided	NN	O	O
into	NN	O	O
three	NN	O	O
motifs	NN	O	O
,	NN	O	O
A	NN	O	O
,	NN	O	O
B	NN	O	O
and	NN	O	O
C	NN	O	O
,	NN	O	O
each	NN	O	O
of	NN	O	O
which	NN	O	O
influences	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
tax	NN	O	B-protein
activation	NN	O	O
.	NN	O	O

The	NN	O	O
A	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeat	NN	O	I-DNA
has	NN	O	O
strong	NN	O	O
homology	NN	O	O
with	NN	O	O
previously	NN	O	O
described	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP-2	NN	O	I-protein
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
AP-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
present	NN	O	O
in	NN	O	O
T-lymphocytes	NN	O	B-cell_type
and	NN	O	O
that	NN	O	O
cellular	NN	O	O
factors	NN	O	O
from	NN	O	O
both	NN	O	O
non-transformed	NN	O	O
and	NN	O	O
transformed	NN	O	O
T-lymphocytes	NN	O	B-cell_type
specifically	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
motif	NN	O	I-DNA
for	NN	O	O
AP-2	NN	O	B-protein
in	NN	O	O
each	NN	O	O
21	NN	O	O
bp	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
AP-2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
LTR	NN	O	I-DNA
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
we	NN	O	O
used	NN	O	O
an	NN	O	O
AP-2	NN	O	B-DNA
cDNA	NN	O	I-DNA
in	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
transient	NN	O	O
expression	NN	O	O
assays	NN	O	O
.	NN	O	O

Gel	NN	O	O
retardation	NN	O	O
and	NN	O	O
methylation	NN	O	O
interference	NN	O	O
studies	NN	O	O
revealed	NN	O	O
that	NN	O	O
bacterially	NN	O	B-protein
produced	NN	O	I-protein
AP-2	NN	O	I-protein
bound	NN	O	O
specifically	NN	O	O
and	NN	O	O
with	NN	O	O
high	NN	O	O
affinity	NN	O	O
to	NN	O	O
all	NN	O	O
three	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeats	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
that	NN	O	O
it	NN	O	O
required	NN	O	O
the	NN	O	O
core	NN	O	B-DNA
sequence	NN	O	I-DNA
AGGC	NN	O	O
for	NN	O	O
specific	NN	O	O
binding	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
AP-2	NN	O	B-protein
prevented	NN	O	O
the	NN	O	O
subsequent	NN	O	O
binding	NN	O	O
of	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
CREB/ATF	NN	O	B-protein
family	NN	O	I-protein
to	NN	O	O
an	NN	O	O
adjacent	NN	O	B-DNA
regulatory	NN	O	I-DNA
motif	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
an	NN	O	O
AP-2	NN	O	B-DNA
expression	NN	O	I-DNA
construct	NN	O	I-DNA
into	NN	O	O
T-lymphocytes	NN	O	B-cell_type
activated	NN	O	O
gene	NN	O	O
expression	NN	O	O
from	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

At	NN	O	O
least	NN	O	O
two	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeats	NN	O	I-DNA
were	NN	O	O
required	NN	O	O
for	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
AP-2	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
AP-2	NN	O	B-DNA
consensus	NN	O	I-DNA
binding	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
21	NN	O	B-DNA
bp	NN	O	I-DNA
repeats	NN	O	I-DNA
eliminate	NN	O	O
this	NN	O	O
activation	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
dependent	NN	O	O
on	NN	O	O
NFAT-1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
function	NN	O	O
of	NN	O	O
a	NN	O	O
putative	NN	O	B-DNA
NFAT-1	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
enhancer	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
analyzed	NN	O	O
.	NN	O	O

Activation	NN	O	O
by	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
is	NN	O	O
minimal	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
putative	NN	O	B-DNA
NFAT-1	NN	O	I-DNA
region	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
anti-CD3	NN	O	B-protein
or	NN	O	O
to	NN	O	O
mitogens	NN	O	B-protein
in	NN	O	O
T-cell	NN	O	B-cell_line
,	NN	O	O
B-cell	NN	O	B-cell_line
,	NN	O	O
or	NN	O	O
monocyte/macrophage	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
nor	NN	O	O
is	NN	O	O
it	NN	O	O
a	NN	O	O
cis-acting	NN	O	B-DNA
negative	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
OTF-2	NN	O	B-protein
positively	NN	O	O
regulates	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
transplantation	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
DRA	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
promoter	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	B-DNA
histocompatibility	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
DRA	NN	O	B-DNA
contains	NN	O	O
an	NN	O	O
octamer	NN	O	B-DNA
element	NN	O	I-DNA
(	NN	O	O
ATTTGCAT	NN	O	O
)	NN	O	O
that	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
efficient	NN	O	O
DRA	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Several	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
known	NN	O	O
to	NN	O	O
bind	NN	O	O
this	NN	O	O
sequence	NN	O	O
.	NN	O	O

The	NN	O	O
best	NN	O	O
characterized	NN	O	O
are	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
OTF-2	NN	O	I-protein
and	NN	O	O
the	NN	O	O
ubiquitous	NN	O	O
OTF-1	NN	O	B-protein
.	NN	O	O

This	NN	O	O
report	NN	O	O
directly	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
OTF-2	NN	O	B-protein
but	NN	O	O
not	NN	O	O
OTF-1	NN	O	B-protein
regulates	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
analysis	NN	O	O
using	NN	O	O
protein	NN	O	O
fractions	NN	O	O
enriched	NN	O	O
for	NN	O	O
the	NN	O	O
octamer-binding	NN	O	O
protein	NN	O	O
OTF-2	NN	O	B-protein
demonstrate	NN	O	O
a	NN	O	O
positive	NN	O	O
functional	NN	O	O
role	NN	O	O
for	NN	O	O
OTF-2	NN	O	B-protein
in	NN	O	O
DRA	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
OTF-1	NN	O	B-protein
-enriched	NN	O	O
protein	NN	O	O
fractions	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
DRA	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
although	NN	O	O
it	NN	O	O
functionally	NN	O	O
enhanced	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
another	NN	O	O
gene	NN	O	O
.	NN	O	O

Recombinant	NN	O	B-protein
OTF-2	NN	O	I-protein
protein	NN	O	I-protein
produced	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription/translation	NN	O	O
could	NN	O	O
also	NN	O	O
enhance	NN	O	O
DRA	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

In	NN	O	O
vivo	NN	O	O
transient	NN	O	O
transfection	NN	O	O
studies	NN	O	O
utilizing	NN	O	O
an	NN	O	O
OTF-2	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
similar	NN	O	O
findings	NN	O	O
:	NN	O	O
that	NN	O	O
OTF-2	NN	O	B-protein
protein	NN	O	I-protein
enhanced	NN	O	O
DRA	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
this	NN	O	O
effect	NN	O	O
requires	NN	O	O
an	NN	O	O
intact	NN	O	O
octamer	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

Together	NN	O	O
these	NN	O	O
results	NN	O	O
constitute	NN	O	O
the	NN	O	O
first	NN	O	O
direct	NN	O	O
evidence	NN	O	O
of	NN	O	O
a	NN	O	O
positive	NN	O	O
role	NN	O	O
for	NN	O	O
the	NN	O	O
lymphoid-specific	NN	O	B-protein
octamer-binding	NN	O	I-protein
factor	NN	O	I-protein
in	NN	O	O
DRA	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interferon-gamma	NN	O	B-protein
potentiates	NN	O	O
the	NN	O	O
antiviral	NN	O	O
activity	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
interferon-stimulated	NN	O	B-DNA
genes	NN	O	I-DNA
induced	NN	O	O
by	NN	O	O
interferon-alpha	NN	O	B-cell_line
in	NN	O	I-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
interferon	NN	O	I-protein
(	NN	O	I-protein
IFN-alpha/beta	NN	O	I-protein
)	NN	O	I-protein
to	NN	O	I-protein
specific	NN	O	I-protein
receptors	NN	O	I-protein
results	NN	O	O
in	NN	O	O
the	NN	O	O
rapid	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
,	NN	O	O
independent	NN	O	O
of	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
,	NN	O	O
of	NN	O	O
IFN-alpha-stimulated	NN	O	B-DNA
genes	NN	O	I-DNA
(	NN	O	O
ISGs	NN	O	B-DNA
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
and	NN	O	O
HeLa	NN	O	B-cell_line
and	NN	O	O
Daudi	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
ISGF3	NN	O	B-protein
(	NN	O	O
IFN-stimulated	NN	O	B-protein
gene	NN	O	I-protein
factor	NN	O	I-protein
3	NN	O	I-protein
)	NN	O	O
to	NN	O	O
the	NN	O	O
conserved	NN	O	O
IFN-stimulated	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
ISRE	NN	O	B-DNA
)	NN	O	O
results	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

This	NN	O	O
factor	NN	O	O
is	NN	O	O
composed	NN	O	O
of	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
ISGF3	NN	O	B-protein
gamma	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
normally	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
,	NN	O	O
and	NN	O	O
other	NN	O	O
IFN-alpha-activated	NN	O	B-protein
proteins	NN	O	I-protein
which	NN	O	O
preexist	NN	O	O
as	NN	O	O
latent	NN	O	B-protein
cytoplasmic	NN	O	I-protein
precursors	NN	O	I-protein
(	NN	O	O
ISGF3	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
ISG	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_line
U937	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
differs	NN	O	O
from	NN	O	O
most	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
previously	NN	O	O
examined	NN	O	O
.	NN	O	O

U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
express	NN	O	O
both	NN	O	O
type	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
type	NN	O	I-protein
II	NN	O	I-protein
IFN	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
but	NN	O	O
only	NN	O	O
IFN-alpha	NN	O	B-protein
is	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
antiviral	NN	O	O
protection	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Pretreatment	NN	O	O
with	NN	O	O
IFN-gamma	NN	O	B-protein
potentiates	NN	O	O
the	NN	O	O
IFN-alpha	NN	O	B-protein
-induced	NN	O	O
protection	NN	O	O
,	NN	O	O
but	NN	O	O
IFN-gamma	NN	O	B-protein
alone	NN	O	O
does	NN	O	O
not	NN	O	O
have	NN	O	O
any	NN	O	O
antiviral	NN	O	O
activity	NN	O	O
.	NN	O	O

ISG15	NN	O	B-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
not	NN	O	O
detectable	NN	O	O
before	NN	O	O
6	NN	O	O
h	NN	O	O
of	NN	O	O
IFN-alpha	NN	O	B-protein
treatment	NN	O	O
,	NN	O	O
peaks	NN	O	O
at	NN	O	O
24	NN	O	O
h	NN	O	O
,	NN	O	O
and	NN	O	O
requires	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

Although	NN	O	O
IFN-gamma	NN	O	B-protein
alone	NN	O	O
does	NN	O	O
not	NN	O	O
induce	NN	O	O
ISG	NN	O	O
expression	NN	O	O
,	NN	O	O
IFN-gamma	NN	O	B-protein
pretreatment	NN	O	O
markedly	NN	O	O
increases	NN	O	O
and	NN	O	O
hastens	NN	O	O
ISG	NN	O	O
expression	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
extracts	NN	O	O
assayed	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
ISRE	NN	O	B-protein
binding	NN	O	I-protein
factors	NN	O	I-protein
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
show	NN	O	O
that	NN	O	O
ISGF3	NN	O	B-protein
is	NN	O	O
induced	NN	O	O
by	NN	O	O
IFN-alpha	NN	O	B-protein
within	NN	O	O
6	NN	O	O
h	NN	O	O
from	NN	O	O
undetectable	NN	O	O
basal	NN	O	O
levels	NN	O	O
in	NN	O	O
untreated	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
ISGF3	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
the	NN	O	O
latent	NN	O	O
component	NN	O	O
of	NN	O	O
ISGF3	NN	O	B-protein
,	NN	O	O
occurs	NN	O	O
rapidly	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
ISGF3	NN	O	B-protein
activity	NN	O	O
ultimately	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
ISGF3	NN	O	B-protein
gamma	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
IFN-alpha	NN	O	B-protein
or	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Single	NN	O	O
point	NN	O	O
estimation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
normal	NN	O	O
subjects	NN	O	O
and	NN	O	O
of	NN	O	O
children	NN	O	O
under	NN	O	O
long	NN	O	O
term	NN	O	O
glucocorticoid	NN	O	O
treatment	NN	O	O
.	NN	O	O

A	NN	O	O
single	NN	O	O
point	NN	O	O
assay	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
human	NN	O	O
lymphocytes	NN	O	B-cell_type
based	NN	O	O
on	NN	O	O
the	NN	O	O
measurement	NN	O	O
of	NN	O	O
specific	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
has	NN	O	O
been	NN	O	O
developed	NN	O	O
and	NN	O	O
compared	NN	O	O
with	NN	O	O
a	NN	O	O
common	NN	O	O
multi-point	NN	O	O
Scatchard	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
assay	NN	O	O
conditions-concentration	NN	O	O
of	NN	O	O
the	NN	O	O
ligand	NN	O	O
20	NN	O	O
nmol/l	NN	O	O
,	NN	O	O
incubation	NN	O	O
time	NN	O	O
2	NN	O	O
h	NN	O	O
and	NN	O	O
the	NN	O	O
cell	NN	O	O
count	NN	O	O
2-6	NN	O	O
mil	NN	O	O
.	NN	O	O

cells/tube	NN	O	O
in	NN	O	O
the	NN	O	O
assay	NN	O	O
volume	NN	O	O
0.25	NN	O	O
ml	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
optimal	NN	O	O
.	NN	O	O

An	NN	O	O
attempt	NN	O	O
was	NN	O	O
also	NN	O	O
undertaken	NN	O	O
to	NN	O	O
use	NN	O	O
a	NN	O	O
cell	NN	O	O
harvester	NN	O	O
for	NN	O	O
the	NN	O	O
separation	NN	O	O
of	NN	O	O
cells	NN	O	O
from	NN	O	O
unbound	NN	O	O
ligand	NN	O	O
.	NN	O	O

Though	NN	O	O
specifically	NN	O	O
bound	NN	O	O
dexamethasone	NN	O	O
measured	NN	O	O
by	NN	O	O
whole-cell	NN	O	O
assay	NN	O	O
and	NN	O	O
that	NN	O	O
using	NN	O	O
cell	NN	O	O
harvester	NN	O	O
correlated	NN	O	O
well	NN	O	O
,	NN	O	O
almost	NN	O	O
by	NN	O	O
one	NN	O	O
order	NN	O	O
lower	NN	O	O
values	NN	O	O
obtained	NN	O	O
with	NN	O	O
the	NN	O	O
latter	NN	O	O
method	NN	O	O
render	NN	O	O
it	NN	O	O
non-applicable	NN	O	O
for	NN	O	O
receptor	NN	O	O
quantitation	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
from	NN	O	O
9	NN	O	O
healthy	NN	O	O
volunteers	NN	O	O
(	NN	O	O
average	NN	O	O
GR	NN	O	B-protein
concentration	NN	O	O
7131	NN	O	O
+/-	NN	O	O
1256	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
correlated	NN	O	O
excellently	NN	O	O
with	NN	O	O
those	NN	O	O
obtained	NN	O	O
by	NN	O	O
the	NN	O	O
Scatchard	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
single	NN	O	O
point	NN	O	O
assay	NN	O	O
has	NN	O	O
been	NN	O	O
also	NN	O	O
applied	NN	O	O
for	NN	O	O
determination	NN	O	O
of	NN	O	O
GH	NN	O	O
in	NN	O	O
10	NN	O	O
children	NN	O	O
treated	NN	O	O
with	NN	O	O
large	NN	O	O
doses	NN	O	O
of	NN	O	O
prednisone	NN	O	O
.	NN	O	O

The	NN	O	O
average	NN	O	O
values	NN	O	O
from	NN	O	O
healthy	NN	O	O
volunteers	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
significantly	NN	O	O
from	NN	O	O
those	NN	O	O
found	NN	O	O
in	NN	O	O
these	NN	O	O
children	NN	O	O
,	NN	O	O
though	NN	O	O
much	NN	O	O
broader	NN	O	O
range	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
3-O-methyl-D-glucose	NN	O	O
uptake	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
from	NN	O	O
normal	NN	O	O
humans	NN	O	O
:	NN	O	O
correlation	NN	O	O
between	NN	O	O
receptor	NN	O	O
level	NN	O	O
and	NN	O	O
hormone	NN	O	O
effect	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
measured	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentration	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
and	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
were	NN	O	O
isolated	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
from	NN	O	O
ten	NN	O	O
healthy	NN	O	O
male	NN	O	O
volunteers	NN	O	O
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
,	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
on	NN	O	O
3-O-methyl-D-glucose	NN	O	O
uptake	NN	O	O
was	NN	O	O
assayed	NN	O	O
in	NN	O	O
the	NN	O	O
corresponding	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
correlated	NN	O	O
with	NN	O	O
those	NN	O	O
in	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
linear	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
cellular	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
and	NN	O	O
glucocorticoid-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
uptake	NN	O	O
of	NN	O	O
3-O-methyl-D-glucose	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

When	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
were	NN	O	O
incubated	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
8-bromo-cAMP	NN	O	O
,	NN	O	O
cellular	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
increased	NN	O	O
and	NN	O	O
a	NN	O	O
more	NN	O	O
pronounced	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
observed	NN	O	O
on	NN	O	O
the	NN	O	O
transport	NN	O	O
of	NN	O	O
3-O-methyl-D-glucose	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
cellular	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
reflect	NN	O	O
in	NN	O	O
vitro	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
from	NN	O	O
healthy	NN	O	O
males	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
individual	NN	O	O
responsiveness	NN	O	O
may	NN	O	O
alter	NN	O	O
upon	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
levels	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
functional	NN	O	O
stimulation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
encoding	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
:	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
formation	NN	O	O
of	NN	O	O
AP1	NN	O	B-protein
transcriptional	NN	O	I-protein
complex	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
was	NN	O	O
investigated	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
lipopolysaccharide	NN	O	O
induced	NN	O	O
strongly	NN	O	O
both	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
AP1	NN	O	B-protein
formation	NN	O	O
.	NN	O	O

Interferon	NN	O	B-protein
gamma	NN	O	I-protein
activated	NN	O	O
strongly	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
weakly	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
AP1	NN	O	B-protein
.	NN	O	O

Tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
induced	NN	O	O
slightly	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
had	NN	O	O
almost	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
AP1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
differences	NN	O	O
in	NN	O	O
functional	NN	O	O
responses	NN	O	O
elicited	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
by	NN	O	O
all	NN	O	O
three	NN	O	O
factors	NN	O	B-protein
may	NN	O	O
be	NN	O	O
dependent	NN	O	O
on	NN	O	O
different	NN	O	O
routes	NN	O	O
on	NN	O	O
nuclear	NN	O	O
signalling	NN	O	O
employed	NN	O	O
by	NN	O	O
the	NN	O	O
factors	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
production	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
:	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
activation	NN	O	O
signal	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
downregulate	NN	O	O
interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
production	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
two	NN	O	O
different	NN	O	O
mechanims	NN	O	O
:	NN	O	O
direct	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
transcription	NN	O	O
and	NN	O	O
destabilization	NN	O	O
of	NN	O	O
the	NN	O	O
preformed	NN	O	O
interleukin-1	NN	O	B-RNA
beta	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Now	NN	O	O
we	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte	NN	O	O
activating	NN	O	O
signal	NN	O	O
on	NN	O	O
these	NN	O	O
two	NN	O	O
inhibitory	NN	O	O
mechanims	NN	O	O
.	NN	O	O

When	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
were	NN	O	O
preincubated	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
for	NN	O	O
1	NN	O	O
hour	NN	O	O
and	NN	O	O
then	NN	O	O
stimulated	NN	O	O
either	NN	O	O
with	NN	O	O
bacterial	NN	O	O
lipopolysaccharide	NN	O	O
or	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
dexamethasone	NN	O	O
inhibited	NN	O	O
the	NN	O	O
lipopolysaccharide-induced	NN	O	O
interleukin-1	NN	O	B-protein
beta	NN	O	I-protein
protein	NN	O	O
production	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
phorbol	NN	O	O
myristate-induced	NN	O	O
production	NN	O	O
was	NN	O	O
increased	NN	O	O
3-10	NN	O	O
fold	NN	O	O
.	NN	O	O

This	NN	O	O
difference	NN	O	O
was	NN	O	O
also	NN	O	O
seen	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	O
level	NN	O	O
.	NN	O	O

When	NN	O	O
dexamethasone	NN	O	O
was	NN	O	O
added	NN	O	O
to	NN	O	O
the	NN	O	O
cultures	NN	O	O
3	NN	O	O
hours	NN	O	O
after	NN	O	O
the	NN	O	O
stimulators	NN	O	O
,	NN	O	O
it	NN	O	O
clearly	NN	O	O
decreased	NN	O	O
the	NN	O	O
interleukin-1	NN	O	B-RNA
beta	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
regardless	NN	O	O
of	NN	O	O
the	NN	O	O
stimulator	NN	O	O
used	NN	O	O
(	NN	O	O
although	NN	O	O
the	NN	O	O
effect	NN	O	O
was	NN	O	O
clearly	NN	O	O
weaker	NN	O	O
on	NN	O	O
the	NN	O	O
PMA-induced	NN	O	B-RNA
mRNA	NN	O	I-RNA
)	NN	O	O
.	NN	O	O

Thus	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
phorbol	NN	O	O
myristate-induced	NN	O	O
signal	NN	O	O
(	NN	O	O
prolonged	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
activation	NN	O	O
?	NN	O	O
)	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
inhibited	NN	O	O
by	NN	O	O
prior	NN	O	O
incubation	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
it	NN	O	O
also	NN	O	O
protects	NN	O	O
the	NN	O	O
induced	NN	O	B-RNA
mRNA	NN	O	I-RNA
for	NN	O	O
the	NN	O	O
degradative	NN	O	O
action	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
elements	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Induction	NN	O	O
requirements	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Prior	NN	O	O
studies	NN	O	O
have	NN	O	O
identified	NN	O	O
several	NN	O	O
elements	NN	O	O
that	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
stimulated	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
Jurkat	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
sites	NN	O	O
and	NN	O	O
their	NN	O	O
corresponding	NN	O	O
nuclear	NN	O	O
binding	NN	O	O
factors	NN	O	O
include	NN	O	O
:	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
,	NN	O	O
AP-3	NN	O	B-protein
,	NN	O	O
OCT-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

The	NN	O	O
latter	NN	O	O
``	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
for	NN	O	I-protein
activated	NN	O	I-protein
T	NN	O	I-protein
cells	NN	O	I-protein
``	NN	O	O
likely	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
tissue	NN	O	O
specificity	NN	O	O
of	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
studied	NN	O	O
these	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
human	NN	O	O
blood	NN	O	O
to	NN	O	O
verify	NN	O	O
their	NN	O	O
presence	NN	O	O
in	NN	O	O
a	NN	O	O
physiologic	NN	O	O
setting	NN	O	O
and	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
signals	NN	O	O
that	NN	O	O
stimulate	NN	O	O
factor	NN	O	O
activity	NN	O	O
.	NN	O	O

All	NN	O	O
factors	NN	O	O
are	NN	O	O
induced	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
upon	NN	O	O
activation	NN	O	O
with	NN	O	O
mitogens	NN	O	B-protein
but	NN	O	O
not	NN	O	O
with	NN	O	O
exogenous	NN	O	B-protein
IL-2	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
signaling	NN	O	O
requirements	NN	O	O
and	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
inhibitors	NN	O	O
differ	NN	O	O
considerably	NN	O	O
.	NN	O	O

Only	NN	O	O
the	NN	O	O
activities	NN	O	O
for	NN	O	O
NF-AT	NN	O	B-DNA
and	NN	O	I-DNA
AP-1	NN	O	I-DNA
sites	NN	O	I-DNA
require	NN	O	O
two	NN	O	O
signals	NN	O	O
for	NN	O	O
optimal	NN	O	O
induction	NN	O	O
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
PMA	NN	O	O
plus	NN	O	O
either	NN	O	O
lectin	NN	O	B-protein
or	NN	O	O
antibody	NN	O	B-protein
to	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
or	NN	O	O
CD28	NN	O	B-protein
surface	NN	O	I-protein
molecules	NN	O	I-protein
.	NN	O	O

Other	NN	O	O
factors	NN	O	O
are	NN	O	O
induced	NN	O	O
by	NN	O	O
lectin	NN	O	B-protein
,	NN	O	O
antibody	NN	O	B-protein
,	NN	O	O
and/or	NN	O	O
PMA	NN	O	O
alone	NN	O	O
.	NN	O	O

After	NN	O	O
appropriate	NN	O	O
stimulation	NN	O	O
,	NN	O	O
both	NN	O	O
NF-AT	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
are	NN	O	O
peculiarly	NN	O	O
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
inhibitor	NN	O	O
anisomycin	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
correlate	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
and	NN	O	O
AP-1	NN	O	B-protein
in	NN	O	O
gel	NN	O	O
shift	NN	O	O
assays	NN	O	O
with	NN	O	O
the	NN	O	O
two	NN	O	O
signals	NN	O	O
requirements	NN	O	O
for	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
stably	NN	O	O
expressing	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
lymphotropic	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
constitutive	NN	O	O
Tax	NN	O	B-protein
expression	NN	O	O
on	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
with	NN	O	O
its	NN	O	O
recognition	NN	O	B-DNA
sequence	NN	O	I-DNA
and	NN	O	O
on	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
T	NN	O	B-cell_line
lymphoid	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
19D	NN	O	B-cell_line
and	NN	O	O
9J	NN	O	B-cell_line
)	NN	O	O
stably	NN	O	O
transformed	NN	O	O
with	NN	O	O
a	NN	O	O
Tax	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
.	NN	O	O

Tax	NN	O	B-cell_line
expressing	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
contained	NN	O	O
a	NN	O	O
constitutive	NN	O	O
level	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
detectable	NN	O	O
by	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
and	NN	O	O
uv	NN	O	O
cross-linking	NN	O	O
using	NN	O	O
a	NN	O	O
palindromic	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
probe	NN	O	I-DNA
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
interferon	NN	O	B-DNA
beta	NN	O	I-DNA
PRDII	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
Jurkat	NN	O	O
and	NN	O	O
NC2.10	NN	O	O
induction	NN	O	O
with	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
new	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
of	NN	O	O
85	NN	O	B-protein
,	NN	O	I-protein
75	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
54	NN	O	I-protein
kDa	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
Tax	NN	O	B-cell_line
expressing	NN	O	I-cell_line
cells	NN	O	I-cell_line
the	NN	O	O
85-kDa	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
a	NN	O	O
92-kDa	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
were	NN	O	O
constitutively	NN	O	O
induced	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
19D	NN	O	B-cell_line
and	NN	O	O
9J	NN	O	B-cell_line
resulted	NN	O	O
in	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B-dependent	NN	O	I-DNA
granulocyte-macrophage	NN	O	I-DNA
colony	NN	O	I-DNA
stimulating	NN	O	I-DNA
factor	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
increased	NN	O	O
basal	NN	O	O
level	NN	O	O
expression	NN	O	O
of	NN	O	O
transfected	NN	O	O
NF-kappa	NN	O	B-DNA
B-regulated	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

Nonetheless	NN	O	O
transcription	NN	O	O
of	NN	O	O
both	NN	O	O
the	NN	O	O
endogenous	NN	O	O
and	NN	O	O
the	NN	O	O
transfected	NN	O	O
gene	NN	O	O
was	NN	O	O
inducible	NN	O	O
by	NN	O	O
PMA	NN	O	O
treatment	NN	O	O
.	NN	O	O

Tax	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
may	NN	O	O
alter	NN	O	O
the	NN	O	O
stoichiometry	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
thus	NN	O	O
change	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B-regulated	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
NF-GM2	NN	O	B-protein
that	NN	O	O
interacts	NN	O	O
with	NN	O	O
a	NN	O	O
regulatory	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
essential	NN	O	O
for	NN	O	O
its	NN	O	O
induction	NN	O	O
in	NN	O	O
responses	NN	O	O
to	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
:	NN	O	O
purification	NN	O	O
from	NN	O	O
human	NN	O	O
T-cell	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
line	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
similarity	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
antigen	NN	O	O
,	NN	O	O
lectin	NN	O	B-protein
,	NN	O	O
or	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
phorbol-12-myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
(	NN	O	O
A23187	NN	O	O
)	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
genes	NN	O	O
for	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
lymphokines	NN	O	B-protein
,	NN	O	O
including	NN	O	O
granulocyte-macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
GM-CSF	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrated	NN	O	O
in	NN	O	O
earlier	NN	O	O
studies	NN	O	O
that	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
promoter	NN	O	I-DNA
at	NN	O	O
positions	NN	O	O
between	NN	O	O
-95	NN	O	B-DNA
and	NN	O	I-DNA
-73	NN	O	I-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
PMA/A23187	NN	O	O
.	NN	O	O

This	NN	O	O
region	NN	O	O
contains	NN	O	O
two	NN	O	O
DNA-binding	NN	O	B-DNA
motifs	NN	O	I-DNA
,	NN	O	O
GM2	NN	O	B-DNA
and	NN	O	O
GC-box	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
GM2	NN	O	B-DNA
sequence	NN	O	I-DNA
(	NN	O	O
GGTAGTTCCC	NN	O	O
)	NN	O	O
is	NN	O	O
recognized	NN	O	O
by	NN	O	O
an	NN	O	O
inducible	NN	O	B-protein
factor	NN	O	I-protein
NF-GM2	NN	O	I-protein
;	NN	O	O
the	NN	O	O
other	NN	O	O
(	NN	O	O
CCGCCC	NN	O	O
)	NN	O	O
by	NN	O	O
constitutive	NN	O	B-protein
factors	NN	O	I-protein
A1	NN	O	B-protein
,	NN	O	O
A2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
B	NN	O	B-protein
.	NN	O	O

To	NN	O	O
elucidate	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-protein
gene	NN	O	O
activation	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
purified	NN	O	O
the	NN	O	O
inducible	NN	O	B-protein
factor	NN	O	I-protein
NF-GM2	NN	O	I-protein
from	NN	O	O
the	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
of	NN	O	O
stimulated	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
specific	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

The	NN	O	O
purified	NN	O	O
NF-GM2	NN	O	B-protein
consists	NN	O	O
of	NN	O	O
50	NN	O	O
(	NN	O	O
p50	NN	O	B-protein
)	NN	O	O
and	NN	O	O
65	NN	O	O
kDa	NN	O	O
(	NN	O	O
p65	NN	O	B-protein
)	NN	O	O
polypeptides	NN	O	O
and	NN	O	O
has	NN	O	O
a	NN	O	O
binding	NN	O	O
activity	NN	O	O
specific	NN	O	O
for	NN	O	O
both	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-protein
and	NN	O	O
immunoglobulin	NN	O	B-DNA
kappa	NN	O	I-DNA
(	NN	O	I-DNA
GGAAAGTCCC	NN	O	I-DNA
)	NN	O	I-DNA
enhancers	NN	O	I-DNA
.	NN	O	O

Electrophoretically	NN	O	O
purified	NN	O	O
p50	NN	O	B-protein
alone	NN	O	O
can	NN	O	O
form	NN	O	O
a	NN	O	O
protein-DNA	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
but	NN	O	O
in	NN	O	O
the	NN	O	O
mixture	NN	O	O
,	NN	O	O
p50	NN	O	B-protein
associates	NN	O	O
preferentially	NN	O	O
with	NN	O	O
p65	NN	O	B-protein
to	NN	O	O
form	NN	O	O
the	NN	O	O
NF-GM2	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
p65	NN	O	B-protein
gave	NN	O	O
per	NN	O	O
se	NN	O	O
,	NN	O	O
with	NN	O	O
low	NN	O	O
affinity	NN	O	O
,	NN	O	O
a	NN	O	O
protein-DNA	NN	O	B-protein
complex	NN	O	I-protein
that	NN	O	O
migrated	NN	O	O
more	NN	O	O
slowly	NN	O	O
than	NN	O	O
native	NN	O	O
NF-GM2	NN	O	B-protein
complex	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
an	NN	O	O
antiserum	NN	O	O
against	NN	O	O
KBF1	NN	O	B-protein
(	NN	O	O
identical	NN	O	O
to	NN	O	O
50	NN	O	B-protein
kDa	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
reacted	NN	O	O
with	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
of	NN	O	O
NF-GM2	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
NF-GM2	NN	O	B-protein
polypeptide	NN	O	I-protein
can	NN	O	O
not	NN	O	O
be	NN	O	O
immunologically	NN	O	O
differentiated	NN	O	O
from	NN	O	O
the	NN	O	O
50	NN	O	B-protein
kDa	NN	O	I-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
purified	NN	O	O
NF-GM2	NN	O	B-protein
activated	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
while	NN	O	O
it	NN	O	O
failed	NN	O	O
to	NN	O	O
stimulate	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
harboring	NN	O	O
the	NN	O	O
GM2	NN	O	O
sequence	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
mechanism	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
through	NN	O	O
the	NN	O	O
GM2/GC-box	NN	O	B-DNA
sequence	NN	O	I-DNA
is	NN	O	O
different	NN	O	O
from	NN	O	O
that	NN	O	O
of	NN	O	O
genes	NN	O	O
carrying	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
alone	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
an	NN	O	O
immediate-early	NN	O	B-DNA
gene	NN	O	I-DNA
induced	NN	O	O
in	NN	O	O
adherent	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
that	NN	O	O
encodes	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
-like	NN	O	O
activity	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
cDNAs	NN	O	B-DNA
representing	NN	O	O
mRNAs	NN	O	B-RNA
that	NN	O	O
are	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
following	NN	O	O
adherence	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
induced	NN	O	O
transcripts	NN	O	O
(	NN	O	O
MAD-3	NN	O	B-RNA
)	NN	O	O
encodes	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
317	NN	O	O
amino	NN	O	O
acids	NN	O	O
with	NN	O	O
one	NN	O	O
domain	NN	O	O
containing	NN	O	O
five	NN	O	B-protein
tandem	NN	O	I-protein
repeats	NN	O	I-protein
of	NN	O	O
the	NN	O	O
cdc10/ankyrin	NN	O	B-protein
motif	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
60	NN	O	O
%	NN	O	O
similar	NN	O	O
(	NN	O	O
46	NN	O	O
%	NN	O	O
identical	NN	O	O
)	NN	O	O
to	NN	O	O
the	NN	O	O
ankyrin	NN	O	B-protein
repeat	NN	O	I-protein
region	NN	O	I-protein
of	NN	O	O
the	NN	O	O
precursor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/KBF1	NN	O	I-protein
p50	NN	O	I-protein
.	NN	O	O

The	NN	O	O
C-terminus	NN	O	B-protein
has	NN	O	O
a	NN	O	O
putative	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
phosphorylation	NN	O	I-protein
site	NN	O	I-protein
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
translated	NN	O	O
MAD-3	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
specifically	NN	O	O
inhibit	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
p50/p65	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
complex	NN	O	I-protein
but	NN	O	O
not	NN	O	O
that	NN	O	O
of	NN	O	O
the	NN	O	O
p50/p50	NN	O	B-protein
KBF1	NN	O	I-protein
factor	NN	O	I-protein
or	NN	O	O
of	NN	O	O
other	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
MAD-3	NN	O	B-DNA
cDNA	NN	O	I-DNA
encodes	NN	O	O
an	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-like	NN	O	I-protein
protein	NN	O	I-protein
that	NN	O	O
is	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
responses	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
including	NN	O	O
adhesion-dependent	NN	O	O
pathways	NN	O	O
of	NN	O	O
monocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cortivazol	NN	O	O
mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-RNA
receptor	NN	O	I-RNA
messenger	NN	O	I-RNA
ribonucleic	NN	O	I-RNA
acid	NN	O	I-RNA
in	NN	O	O
wild-type	NN	O	B-cell_line
and	NN	O	O
dexamethasone-resistant	NN	O	B-cell_line
human	NN	O	I-cell_line
leukemic	NN	O	I-cell_line
(	NN	O	I-cell_line
CEM	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Cortivazol	NN	O	O
is	NN	O	O
a	NN	O	O
phenylpyrazolo	NN	O	O
glucocorticoid	NN	O	O
of	NN	O	O
high	NN	O	O
potency	NN	O	O
and	NN	O	O
unusual	NN	O	O
structure	NN	O	O
.	NN	O	O

In	NN	O	O
both	NN	O	O
wild-type	NN	O	B-cell_line
and	NN	O	O
highly	NN	O	O
dexamethasone	NN	O	B-cell_line
(	NN	O	I-cell_line
dex	NN	O	I-cell_line
)	NN	O	I-cell_line
-resistant	NN	O	I-cell_line
clones	NN	O	I-cell_line
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
CEM	NN	O	I-cell_line
,	NN	O	O
exposure	NN	O	O
to	NN	O	O
cortivazol	NN	O	O
leads	NN	O	O
to	NN	O	O
cell	NN	O	O
death	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
recently	NN	O	O
that	NN	O	O
in	NN	O	O
wild-type	NN	O	B-cell_line
CEM	NN	O	I-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
a	NN	O	O
dex-resistant	NN	O	B-cell_line
,	NN	O	I-cell_line
glucocorticoid	NN	O	I-cell_line
receptor	NN	O	I-cell_line
(	NN	O	I-cell_line
GR	NN	O	I-cell_line
)	NN	O	I-cell_line
-defective	NN	O	I-cell_line
clone	NN	O	I-cell_line
ICR-27	NN	O	B-cell_line
TK-3	NN	O	I-cell_line
,	NN	O	O
dex	NN	O	O
induces	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

To	NN	O	O
test	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
cortivazol	NN	O	O
acts	NN	O	O
in	NN	O	O
dex-resistant	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
making	NN	O	O
use	NN	O	O
of	NN	O	O
the	NN	O	O
residual	NN	O	O
GR	NN	O	B-protein
found	NN	O	O
there	NN	O	O
,	NN	O	O
wild-type	NN	O	O
and	NN	O	O
dex-resistant	NN	O	B-cell_line
clones	NN	O	I-cell_line
were	NN	O	O
treated	NN	O	O
with	NN	O	O
various	NN	O	O
concentrations	NN	O	O
of	NN	O	O
cortivazol	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
studied	NN	O	O
.	NN	O	O

Cortivazol	NN	O	O
significantly	NN	O	O
induced	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
normal	NN	O	B-cell_line
CEM-C7	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
two	NN	O	O
classes	NN	O	O
of	NN	O	O
dex-resistant	NN	O	B-cell_line
clones	NN	O	I-cell_line
,	NN	O	O
although	NN	O	O
the	NN	O	O
dex-resistant	NN	O	B-cell_line
clones	NN	O	I-cell_line
needed	NN	O	O
at	NN	O	O
least	NN	O	O
10	NN	O	O
times	NN	O	O
more	NN	O	O
cortivazol	NN	O	O
than	NN	O	O
the	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
significant	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
.	NN	O	O

Increased	NN	O	O
levels	NN	O	O
of	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
were	NN	O	O
noticed	NN	O	O
as	NN	O	O
early	NN	O	O
as	NN	O	O
3	NN	O	O
h	NN	O	O
after	NN	O	O
treatment	NN	O	O
.	NN	O	O

A	NN	O	O
general	NN	O	O
correlation	NN	O	O
between	NN	O	O
induction	NN	O	O
of	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
lysis	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
and	NN	O	O
dex-resistant	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
found	NN	O	O
.	NN	O	O

Positive	NN	O	O
induction	NN	O	O
of	NN	O	O
GR	NN	O	B-RNA
mRNA	NN	O	I-RNA
might	NN	O	O
be	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
earliest	NN	O	O
crucial	NN	O	O
steps	NN	O	O
in	NN	O	O
the	NN	O	O
lysis	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_line
and	NN	O	I-cell_line
dex-resistant	NN	O	I-cell_line
CEM	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
or	NN	O	O
might	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
marker	NN	O	O
for	NN	O	O
the	NN	O	O
process	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
lysis	NN	O	O
pathway	NN	O	O
in	NN	O	O
the	NN	O	O
dex-resistant	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
defective	NN	O	O
in	NN	O	O
that	NN	O	O
dex-resistant	NN	O	B-cell_line
clones	NN	O	I-cell_line
needed	NN	O	O
significantly	NN	O	O
more	NN	O	O
cortivazol	NN	O	O
than	NN	O	O
the	NN	O	O
normal	NN	O	B-cell_line
cells	NN	O	I-cell_line
for	NN	O	O
lysis	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
perpetuated	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
induction	NN	O	O
on	NN	O	O
infection	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Permissiveness	NN	O	O
to	NN	O	O
replication	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
differs	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
is	NN	O	O
poorly	NN	O	O
detected	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Cloned	NN	O	O
,	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
show	NN	O	O
very	NN	O	O
little	NN	O	O
,	NN	O	O
if	NN	O	O
any	NN	O	O
,	NN	O	O
basal	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
low	NN	O	O
nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
a	NN	O	O
potent	NN	O	B-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
fixed	NN	O	O
tissue	NN	O	O
macrophages	NN	O	B-cell_type
express	NN	O	O
detectable	NN	O	O
HIV	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
indicating	NN	O	O
permanent	NN	O	O
virus	NN	O	O
transcription	NN	O	O
.	NN	O	O

One	NN	O	O
explanation	NN	O	O
for	NN	O	O
the	NN	O	O
perpetuation	NN	O	O
of	NN	O	O
virus	NN	O	O
infection	NN	O	O
in	NN	O	O
macrophages	NN	O	B-cell_type
could	NN	O	O
be	NN	O	O
sustained	NN	O	O
nuclear	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
U937	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
is	NN	O	O
fully	NN	O	O
permissive	NN	O	O
to	NN	O	O
HIV	NN	O	O
replication	NN	O	O
,	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
express	NN	O	O
only	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
chronic	NN	O	O
HIV	NN	O	O
infection	NN	O	O
results	NN	O	O
in	NN	O	O
both	NN	O	O
induction	NN	O	O
of	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
with	NN	O	O
antigenic	NN	O	O
properties	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
permanently	NN	O	O
increased	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

This	NN	O	O
phenomenon	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
independent	NN	O	O
of	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
HIV	NN	O	O
replication	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
thus	NN	O	O
likely	NN	O	O
to	NN	O	O
explain	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
the	NN	O	O
perpetuation	NN	O	O
of	NN	O	O
HIV	NN	O	O
infection	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Isolation	NN	O	O
of	NN	O	O
a	NN	O	O
rel-related	NN	O	B-DNA
human	NN	O	I-DNA
cDNA	NN	O	I-DNA
that	NN	O	O
potentially	NN	O	O
encodes	NN	O	O
the	NN	O	O
65-kD	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
Science	NN	O	O
1991	NN	O	O
Oct	NN	O	O
4	NN	O	O
;	NN	O	O
254	NN	O	O
(	NN	O	O
5028	NN	O	O
)	NN	O	O
:	NN	O	O
11	NN	O	O
]	NN	O	O

A	NN	O	O
DNA	NN	O	B-DNA
probe	NN	O	I-DNA
that	NN	O	O
spanned	NN	O	O
a	NN	O	O
domain	NN	O	O
conserved	NN	O	O
among	NN	O	O
the	NN	O	O
proto-oncogene	NN	O	B-DNA
c-rel	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-protein
morphogen	NN	O	I-protein
dorsal	NN	O	I-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
generated	NN	O	O
from	NN	O	O
Jurkat	NN	O	B-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
complementary	NN	O	I-DNA
DNA	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
(	NN	O	O
PCR	NN	O	O
)	NN	O	O
and	NN	O	O
degenerate	NN	O	O
oligonucleotides	NN	O	O
.	NN	O	O

This	NN	O	O
probe	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
a	NN	O	O
rel-related	NN	O	B-DNA
complementary	NN	O	I-DNA
DNA	NN	O	I-DNA
that	NN	O	O
hybridized	NN	O	O
to	NN	O	O
a	NN	O	O
2.6-kilobase	NN	O	B-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
present	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
and	NN	O	O
translation	NN	O	O
of	NN	O	O
the	NN	O	O
complementary	NN	O	B-DNA
DNA	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
a	NN	O	O
protein	NN	O	O
with	NN	O	O
an	NN	O	O
apparent	NN	O	O
molecular	NN	O	O
size	NN	O	O
of	NN	O	O
65	NN	O	B-protein
kilodaltons	NN	O	I-protein
(	NN	O	O
kD	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
translated	NN	O	O
protein	NN	O	O
showed	NN	O	O
weak	NN	O	O
DNA	NN	O	O
binding	NN	O	O
with	NN	O	O
a	NN	O	O
specificity	NN	O	O
for	NN	O	O
the	NN	O	O
kappa	NN	O	O
B	NN	O	O
binding	NN	O	O
motif	NN	O	O
.	NN	O	O

This	NN	O	O
protein-DNA	NN	O	B-protein
complex	NN	O	I-protein
comigrated	NN	O	O
with	NN	O	O
the	NN	O	O
complex	NN	O	O
obtained	NN	O	O
with	NN	O	O
the	NN	O	O
purified	NN	O	B-protein
human	NN	O	I-protein
p65	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
subunit	NN	O	I-protein
and	NN	O	O
binding	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B-alpha	NN	O	I-protein
and	NN	O	O
-beta	NN	O	O
proteins	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
65-kD	NN	O	B-protein
protein	NN	O	I-protein
associated	NN	O	O
with	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
probe	NN	O	I-DNA
to	NN	O	O
form	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
the	NN	O	O
same	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
as	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B-DNA	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Therefore	NN	O	O
the	NN	O	O
rel-related	NN	O	B-protein
65-kD	NN	O	I-protein
protein	NN	O	I-protein
may	NN	O	O
represent	NN	O	O
the	NN	O	O
p65	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
active	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
human	NN	O	B-cell_line
myelomonocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
U-937	NN	O	I-cell_line
as	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
studying	NN	O	O
alterations	NN	O	O
in	NN	O	O
steroid-induced	NN	O	O
monokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
:	NN	O	O
marked	NN	O	O
enhancement	NN	O	O
of	NN	O	O
lipopolysaccharide-stimulated	NN	O	B-RNA
interleukin-1	NN	O	I-RNA
beta	NN	O	I-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
levels	NN	O	O
by	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
.	NN	O	O

The	NN	O	O
active	NN	O	O
metabolite	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
[	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
]	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
regulator	NN	O	O
of	NN	O	O
human	NN	O	O
monocyte/macrophage	NN	O	O
function	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

To	NN	O	O
establish	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	O
monocyte	NN	O	O
monokine	NN	O	O
synthesis	NN	O	O
,	NN	O	O
three	NN	O	O
human	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
(	NN	O	O
U-937	NN	O	B-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
HL-60	NN	O	B-cell_line
)	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
:	NN	O	O
1	NN	O	O
)	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
functional	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
receptors	NN	O	O
;	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
accumulation	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-RNA
beta	NN	O	I-RNA
(	NN	O	I-RNA
IL-1	NN	O	I-RNA
beta	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
IL-1	NN	O	B-protein
beta	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
;	NN	O	O
and	NN	O	O
3	NN	O	O
)	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
this	NN	O	O
response	NN	O	O
by	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
.	NN	O	O

All	NN	O	O
three	NN	O	O
cell	NN	O	O
lines	NN	O	O
expressed	NN	O	O
vitamin	NN	O	O
D	NN	O	O
receptor	NN	O	O
and	NN	O	O
had	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
IL-1	NN	O	B-RNA
beta	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
.	NN	O	O

Preincubation	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
augmented	NN	O	O
IL-1	NN	O	B-RNA
beta	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
only	NN	O	O
in	NN	O	O
U-937	NN	O	B-cell_line
and	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	O
.	NN	O	O

From	NN	O	O
these	NN	O	O
data	NN	O	O
,	NN	O	O
and	NN	O	O
taking	NN	O	O
into	NN	O	O
consideration	NN	O	O
their	NN	O	O
state	NN	O	O
of	NN	O	O
differentiation	NN	O	O
and	NN	O	O
relative	NN	O	O
ease	NN	O	O
of	NN	O	O
culture	NN	O	O
,	NN	O	O
U-937	NN	O	B-cell_line
was	NN	O	O
chosen	NN	O	O
over	NN	O	O
HL-60	NN	O	B-cell_line
and	NN	O	O
THP-1	NN	O	B-cell_line
as	NN	O	O
the	NN	O	O
cell	NN	O	O
line	NN	O	O
we	NN	O	O
further	NN	O	O
characterized	NN	O	O
.	NN	O	O

In	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
optimum	NN	O	O
time	NN	O	O
and	NN	O	O
dose	NN	O	O
of	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
were	NN	O	O
determined	NN	O	O
to	NN	O	O
be	NN	O	O
12-24	NN	O	O
h	NN	O	O
at	NN	O	O
a	NN	O	O
receptor	NN	O	O
saturating	NN	O	O
concentration	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
(	NN	O	O
10	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

Preincubation	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
time	NN	O	O
course	NN	O	O
of	NN	O	O
IL-1	NN	O	B-RNA
beta	NN	O	I-RNA
mRNA	NN	O	I-RNA
appearance	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
LPS	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
exposure	NN	O	O
of	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
increased	NN	O	O
by	NN	O	O
200	NN	O	O
%	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
IL-1	NN	O	B-RNA
beta	NN	O	I-RNA
mRNA	NN	O	I-RNA
detected	NN	O	O
and	NN	O	O
decreased	NN	O	O
by	NN	O	O
three	NN	O	O
orders	NN	O	O
of	NN	O	O
magnitude	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
LPS	NN	O	O
required	NN	O	O
to	NN	O	O
achieve	NN	O	O
steady	NN	O	O
state	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
equivalent	NN	O	O
to	NN	O	O
those	NN	O	O
observed	NN	O	O
in	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
not	NN	O	O
preincubated	NN	O	O
with	NN	O	O
the	NN	O	O
hormone.2+o	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
M-CSF	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
M-CSF	NN	O	B-protein
:	NN	O	O
role	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
and	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Macrophage-colony-stimulating	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
M-CSF	NN	O	B-protein
)	NN	O	O
,	NN	O	O
also	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
CSF-1	NN	O	B-protein
,	NN	O	O
regulates	NN	O	O
the	NN	O	O
survival	NN	O	O
,	NN	O	O
growth	NN	O	O
,	NN	O	O
differentiation	NN	O	O
and	NN	O	O
functional	NN	O	O
activity	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
by	NN	O	O
binding	NN	O	O
to	NN	O	O
a	NN	O	O
single	NN	O	O
class	NN	O	O
of	NN	O	O
high-affinity	NN	O	B-protein
cell	NN	O	I-protein
surface	NN	O	I-protein
receptors	NN	O	I-protein
,	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
the	NN	O	O
c-fms	NN	O	B-DNA
protooncogene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
detection	NN	O	O
of	NN	O	O
both	NN	O	O
M-CSF	NN	O	O
and	NN	O	O
c-fms	NN	O	O
expression	NN	O	O
by	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte	NN	O	B-cell_type
lineage	NN	O	I-cell_type
has	NN	O	O
suggested	NN	O	O
that	NN	O	O
M-CSF	NN	O	B-protein
may	NN	O	O
act	NN	O	O
by	NN	O	O
an	NN	O	O
autocrine	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
it	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
that	NN	O	O
M-CSF	NN	O	B-protein
can	NN	O	O
induce	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
its	NN	O	O
own	NN	O	O
gene	NN	O	O
.	NN	O	O

Although	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
M-CSF	NN	O	B-protein
can	NN	O	O
be	NN	O	O
modulated	NN	O	O
by	NN	O	O
regulation	NN	O	O
of	NN	O	O
receptor	NN	O	O
expression	NN	O	O
and	NN	O	O
function	NN	O	O
,	NN	O	O
M-CSF	NN	O	B-protein
responsiveness	NN	O	O
is	NN	O	O
largely	NN	O	O
determined	NN	O	O
at	NN	O	O
a	NN	O	O
postreceptor	NN	O	O
level	NN	O	O
.	NN	O	O

To	NN	O	O
date	NN	O	O
,	NN	O	O
little	NN	O	O
is	NN	O	O
known	NN	O	O
about	NN	O	O
the	NN	O	O
intracellular	NN	O	O
pathway	NN	O	O
of	NN	O	O
M-CSF	NN	O	B-protein
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
therefore	NN	O	O
investigated	NN	O	O
the	NN	O	O
changes	NN	O	O
in	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
activity	NN	O	O
upon	NN	O	O
exposure	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
to	NN	O	O
M-CSF	NN	O	B-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
M-CSF	NN	O	B-protein
activates	NN	O	O
and	NN	O	O
translocates	NN	O	O
PKC	NN	O	B-protein
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
by	NN	O	O
the	NN	O	O
isoquinoline	NN	O	O
derivative	NN	O	O
H7	NN	O	O
abolishes	NN	O	O
induction	NN	O	O
of	NN	O	O
M-CSF	NN	O	B-protein
by	NN	O	O
M-CSF	NN	O	B-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
was	NN	O	O
pertussis-toxin-sensitive	NN	O	O
and	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
detection	NN	O	O
of	NN	O	O
an	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
M-CSF-induced	NN	O	B-cell_line
blood	NN	O	I-cell_line
monocytes	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
exposed	NN	O	O
to	NN	O	O
medium	NN	O	O
treatment	NN	O	O
only	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
M-CSF	NN	O	B-protein
induction	NN	O	O
of	NN	O	O
M-CSF	NN	O	B-protein
involves	NN	O	O
G	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
PKC	NN	O	B-protein
and	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Clone	NN	O	B-DNA
pAT	NN	O	I-DNA
133	NN	O	I-DNA
identifies	NN	O	O
a	NN	O	O
gene	NN	O	O
that	NN	O	O
encodes	NN	O	O
another	NN	O	O
human	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
class	NN	O	O
of	NN	O	O
growth	NN	O	B-DNA
factor-induced	NN	O	I-DNA
genes	NN	O	I-DNA
with	NN	O	O
almost	NN	O	O
identical	NN	O	O
zinc-finger	NN	O	B-protein
domains	NN	O	I-protein
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
structure	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
a	NN	O	O
gene	NN	O	O
represented	NN	O	O
by	NN	O	O
clone	NN	O	B-DNA
pAT	NN	O	I-DNA
133	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
induced	NN	O	O
upon	NN	O	O
transition	NN	O	O
from	NN	O	O
a	NN	O	O
resting	NN	O	O
state	NN	O	O
(	NN	O	O
G0	NN	O	O
)	NN	O	O
through	NN	O	O
the	NN	O	O
early	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
(	NN	O	O
G1	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
pAT	NN	O	B-DNA
133	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
immediately	NN	O	O
induced	NN	O	O
,	NN	O	O
with	NN	O	O
FOS	NN	O	B-protein
-like	NN	O	O
kinetics	NN	O	O
,	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

Primary	NN	O	O
structure	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
encoded	NN	O	O
protein	NN	O	O
contains	NN	O	O
three	NN	O	O
tandem	NN	O	B-protein
zinc-finger	NN	O	I-protein
sequences	NN	O	I-protein
of	NN	O	O
the	NN	O	O
type	NN	O	O
Cys2-Xaa12-His2	NN	O	B-protein
.	NN	O	O

This	NN	O	O
zinc-finger	NN	O	B-protein
region	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
bind	NN	O	O
DNA	NN	O	O
in	NN	O	O
a	NN	O	O
sequence-specific	NN	O	O
manner	NN	O	O
,	NN	O	O
is	NN	O	O
similar	NN	O	O
(	NN	O	O
greater	NN	O	O
than	NN	O	O
80	NN	O	O
%	NN	O	O
on	NN	O	O
the	NN	O	O
amino	NN	O	O
acid	NN	O	O
level	NN	O	O
)	NN	O	O
to	NN	O	O
two	NN	O	O
previously	NN	O	O
described	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
pAT	NN	O	B-protein
225/EGR1	NN	O	I-protein
and	NN	O	O
pAT	NN	O	O
591/EGR2	NN	O	B-protein
.	NN	O	O

Except	NN	O	O
for	NN	O	O
the	NN	O	O
conserved	NN	O	O
zinc-finger	NN	O	B-protein
domains	NN	O	I-protein
,	NN	O	O
the	NN	O	O
amino	NN	O	B-protein
acid	NN	O	I-protein
sequences	NN	O	I-protein
of	NN	O	O
the	NN	O	O
three	NN	O	O
proteins	NN	O	O
are	NN	O	O
distinct	NN	O	O
.	NN	O	O

This	NN	O	O
structural	NN	O	O
similarity	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
pAT	NN	O	B-DNA
133	NN	O	I-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
with	NN	O	O
a	NN	O	O
specific	NN	O	O
biological	NN	O	O
function	NN	O	O
.	NN	O	O

Comparing	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
these	NN	O	O
related	NN	O	O
zinc-finger-encoding	NN	O	B-DNA
genes	NN	O	I-DNA
showed	NN	O	O
coordinate	NN	O	O
induction	NN	O	O
upon	NN	O	O
mitogenic	NN	O	O
stimulation	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
of	NN	O	O
resting	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
upon	NN	O	O
transition	NN	O	O
from	NN	O	O
a	NN	O	O
proliferating	NN	O	O
(	NN	O	O
G1	NN	O	O
)	NN	O	O
to	NN	O	O
a	NN	O	O
resting	NN	O	O
state	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
the	NN	O	O
three	NN	O	O
genes	NN	O	O
were	NN	O	O
differently	NN	O	O
regulated	NN	O	O
.	NN	O	O

In	NN	O	O
human	NN	O	B-RNA
histiocytic	NN	O	I-RNA
U937	NN	O	I-RNA
cells	NN	O	I-RNA
mRNA	NN	O	I-RNA
of	NN	O	O
clone	NN	O	O
pAT	NN	O	O
133	NN	O	O
was	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
,	NN	O	O
whereas	NN	O	O
mRNA	NN	O	B-RNA
of	NN	O	O
pAT	NN	O	B-protein
225/EGR1	NN	O	I-protein
was	NN	O	O
induced	NN	O	O
upon	NN	O	O
induction	NN	O	O
of	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
mRNA	NN	O	O
representing	NN	O	O
pAT	NN	O	O
591/EGR2	NN	O	B-protein
was	NN	O	O
not	NN	O	O
expressed	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

This	NN	O	O
difference	NN	O	O
in	NN	O	O
gene	NN	O	O
regulation	NN	O	O
suggests	NN	O	O
distinct	NN	O	O
biological	NN	O	O
roles	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
for	NN	O	O
the	NN	O	O
respective	NN	O	O
proteins	NN	O	O
.	NN	O	O

-DOCSTART-	O

v-erbA	NN	O	B-protein
overexpression	NN	O	O
is	NN	O	O
required	NN	O	O
to	NN	O	O
extinguish	NN	O	O
c-erbA	NN	O	B-protein
function	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
erbA	NN	O	B-DNA
target	NN	O	I-DNA
gene	NN	O	I-DNA
CAII	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
v-erbA	NN	O	B-protein
oncoprotein	NN	O	I-protein
represents	NN	O	O
a	NN	O	O
retrovirus-transduced	NN	O	O
oncogenic	NN	O	O
version	NN	O	O
of	NN	O	O
the	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
(	NN	O	I-protein
T3/T4	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
c-erbA	NN	O	B-protein
(	NN	O	I-protein
type	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

It	NN	O	O
contributes	NN	O	O
to	NN	O	O
virus-induced	NN	O	O
erythroleukemia	NN	O	O
by	NN	O	O
efficiently	NN	O	O
arresting	NN	O	O
differentiation	NN	O	O
of	NN	O	O
red	NN	O	B-cell_type
cell	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
and	NN	O	O
by	NN	O	O
suppressing	NN	O	O
transcription	NN	O	O
of	NN	O	O
erythrocyte-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
v-erbA	NN	O	B-protein
and	NN	O	O
c-erbA	NN	O	B-protein
bind	NN	O	O
directly	NN	O	O
to	NN	O	O
sequences	NN	O	O
within	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
erythrocyte-specific	NN	O	B-protein
carbonic	NN	O	I-protein
anhydrase	NN	O	I-protein
II	NN	O	I-protein
(	NN	O	O
CAII	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
a	NN	O	O
gene	NN	O	O
whose	NN	O	O
transcription	NN	O	O
is	NN	O	O
efficiently	NN	O	O
suppressed	NN	O	O
by	NN	O	O
v-erbA	NN	O	B-protein
.	NN	O	O

This	NN	O	O
erbA-binding	NN	O	B-DNA
site	NN	O	I-DNA
confers	NN	O	O
thyroid	NN	O	O
hormone	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
transient	NN	O	O
expression	NN	O	O
experiments	NN	O	O
and	NN	O	O
is	NN	O	O
a	NN	O	O
target	NN	O	O
for	NN	O	O
efficient	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
CAII	NN	O	B-DNA
transcription	NN	O	O
by	NN	O	O
the	NN	O	O
v-erbA	NN	O	B-protein
oncoprotein	NN	O	I-protein
.	NN	O	O

In	NN	O	O
stably	NN	O	B-cell_line
transformed	NN	O	I-cell_line
erythroblasts	NN	O	I-cell_line
coexpressing	NN	O	O
the	NN	O	O
v-erbA	NN	O	B-protein
oncoprotein	NN	O	I-protein
and	NN	O	O
the	NN	O	O
c-erbA/T3	NN	O	B-protein
receptor	NN	O	I-protein
at	NN	O	O
an	NN	O	O
approximately	NN	O	O
equimolar	NN	O	O
ratio	NN	O	O
,	NN	O	O
c-erbA	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
dominant	NN	O	O
over	NN	O	O
v-erbA	NN	O	B-protein
.	NN	O	O

T3	NN	O	O
efficiently	NN	O	O
induced	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
thus	NN	O	O
overcoming	NN	O	O
the	NN	O	O
v-erbA	NN	O	B-protein
-mediated	NN	O	O
differentiation	NN	O	O
arrest	NN	O	O
.	NN	O	O

Likewise	NN	O	O
,	NN	O	O
T3	NN	O	O
activated	NN	O	O
CAII	NN	O	B-DNA
transcription	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
transient	NN	O	O
expression	NN	O	O
of	NN	O	O
a	NN	O	O
T3-responsive	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
containing	NN	O	O
the	NN	O	O
CAII-specific	NN	O	B-DNA
erbA-binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
c-erbA	NN	O	B-protein
-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
CAII	NN	O	B-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
could	NN	O	O
only	NN	O	O
be	NN	O	O
suppressed	NN	O	O
by	NN	O	O
very	NN	O	O
high	NN	O	O
amounts	NN	O	O
of	NN	O	O
v-erbA	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
overexpression	NN	O	O
of	NN	O	O
v-erbA	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
its	NN	O	O
function	NN	O	O
as	NN	O	O
an	NN	O	O
oncoprotein	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Anti-CD2	NN	O	B-protein
receptor	NN	O	I-protein
antibodies	NN	O	I-protein
activate	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
CD2	NN	O	B-protein
T	NN	O	I-protein
lymphocyte	NN	O	I-protein
glycoprotein	NN	O	I-protein
surface	NN	O	I-protein
molecule	NN	O	I-protein
mediates	NN	O	O
both	NN	O	O
cell	NN	O	O
to	NN	O	O
cell	NN	O	O
adhesion	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
two	NN	O	O
processes	NN	O	O
that	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
spread	NN	O	O
of	NN	O	O
HIV	NN	O	O
infection	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
chronically	NN	O	B-cell_line
HIV-infected	NN	O	I-cell_line
PBMC	NN	O	I-cell_line
with	NN	O	O
anti-CD2	NN	O	B-protein
mAb	NN	O	I-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
infectious	NN	O	O
virus	NN	O	O
from	NN	O	O
these	NN	O	O
cultures	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
whereby	NN	O	O
anti-CD2	NN	O	B-protein
antibodies	NN	O	I-protein
stimulate	NN	O	O
viral	NN	O	O
production	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
treatment	NN	O	O
of	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
anti-CD2	NN	O	B-protein
antibodies	NN	O	I-protein
results	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
CAT	NN	O	B-protein
assays	NN	O	O
using	NN	O	O
mutated	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat-CAT	NN	O	I-DNA
constructs	NN	O	I-DNA
and	NN	O	O
gel	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
this	NN	O	O
activation	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
studies	NN	O	O
suggest	NN	O	O
that	NN	O	O
interaction	NN	O	O
of	NN	O	O
CD2	NN	O	B-protein
with	NN	O	O
its	NN	O	O
natural	NN	O	O
ligand	NN	O	O
,	NN	O	O
LFA-3	NN	O	B-protein
,	NN	O	O
may	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
regulation	NN	O	O
of	NN	O	O
HIV	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
as	NN	O	O
apparently	NN	O	O
widely	NN	O	O
used	NN	O	O
messengers	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

Hydrogen	NN	O	O
peroxide	NN	O	O
and	NN	O	O
oxygen	NN	O	O
radicals	NN	O	O
are	NN	O	O
agents	NN	O	O
commonly	NN	O	O
produced	NN	O	O
during	NN	O	O
inflammatory	NN	O	O
processes	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
micromolar	NN	O	O
concentrations	NN	O	O
of	NN	O	O
H2O2	NN	O	O
can	NN	O	O
induce	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
replication	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
effect	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
which	NN	O	O
is	NN	O	O
potently	NN	O	O
and	NN	O	O
rapidly	NN	O	O
activated	NN	O	O
by	NN	O	O
an	NN	O	O
H2O2	NN	O	O
treatment	NN	O	O
of	NN	O	O
cells	NN	O	O
from	NN	O	O
its	NN	O	O
inactive	NN	O	B-protein
cytoplasmic	NN	O	I-protein
form	NN	O	I-protein
.	NN	O	O

N-acetyl-L-cysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
well	NN	O	O
characterized	NN	O	O
antioxidant	NN	O	O
which	NN	O	O
counteracts	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediates	NN	O	O
(	NN	O	O
ROI	NN	O	O
)	NN	O	O
in	NN	O	O
living	NN	O	O
cells	NN	O	O
,	NN	O	O
prevented	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
H2O2	NN	O	O
.	NN	O	O

NAC	NN	O	O
and	NN	O	O
other	NN	O	O
thiol	NN	O	O
compounds	NN	O	O
also	NN	O	O
blocked	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
cycloheximide	NN	O	O
,	NN	O	O
double-stranded	NN	O	B-RNA
RNA	NN	O	I-RNA
,	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
active	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
,	NN	O	O
interleukin-1	NN	O	B-protein
,	NN	O	O
lipopolysaccharide	NN	O	O
and	NN	O	O
lectin	NN	O	B-protein
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
diverse	NN	O	O
agents	NN	O	O
thought	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
distinct	NN	O	O
intracellular	NN	O	O
pathways	NN	O	O
might	NN	O	O
all	NN	O	O
act	NN	O	O
through	NN	O	O
a	NN	O	O
common	NN	O	O
mechanism	NN	O	O
involving	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
ROI	NN	O	O
.	NN	O	O

ROI	NN	O	O
appear	NN	O	O
to	NN	O	O
serve	NN	O	O
as	NN	O	O
messengers	NN	O	O
mediating	NN	O	O
directly	NN	O	O
or	NN	O	O
indirectly	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	B-protein
subunit	NN	O	I-protein
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
from	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
belonging	NN	O	O
to	NN	O	O
the	NN	O	O
rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
by	NN	O	O
a	NN	O	O
transdominant	NN	O	B-protein
negative	NN	O	I-protein
mutant	NN	O	I-protein
.	NN	O	O

The	NN	O	O
KBF1	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
A	NN	O	I-DNA
located	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
MHC	NN	O	I-DNA
class	NN	O	I-DNA
I	NN	O	I-DNA
gene	NN	O	I-DNA
H-2Kb	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
the	NN	O	O
p50	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
regulates	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
immune	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
responses	NN	O	O
.	NN	O	O

The	NN	O	O
KBF1/p50	NN	O	B-protein
factor	NN	O	I-protein
binds	NN	O	O
as	NN	O	O
a	NN	O	O
homodimer	NN	O	B-protein
but	NN	O	O
can	NN	O	O
also	NN	O	O
form	NN	O	O
heterodimers	NN	O	B-protein
with	NN	O	O
the	NN	O	O
products	NN	O	O
of	NN	O	O
other	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
family	NN	O	O
,	NN	O	O
like	NN	O	O
the	NN	O	O
c-rel	NN	O	B-DNA
and	NN	O	I-DNA
v-rel	NN	O	I-DNA
(	NN	O	I-DNA
proto	NN	O	I-DNA
)	NN	O	I-DNA
oncogenes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
dimerization	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
KBF1/p50	NN	O	B-protein
is	NN	O	O
contained	NN	O	O
between	NN	O	B-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
201	NN	O	I-protein
and	NN	O	I-protein
367	NN	O	I-protein
.	NN	O	O

A	NN	O	O
mutant	NN	O	O
of	NN	O	O
KBF1/p50	NN	O	B-protein
(	NN	O	O
delta	NN	O	B-protein
SP	NN	O	I-protein
)	NN	O	O
,	NN	O	O
unable	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
DNA	NN	O	O
but	NN	O	O
able	NN	O	O
to	NN	O	O
form	NN	O	O
homo-	NN	O	O
or	NN	O	O
heterodimers	NN	O	B-protein
,	NN	O	O
has	NN	O	O
been	NN	O	O
constructed	NN	O	O
.	NN	O	O

This	NN	O	O
protein	NN	O	O
reduces	NN	O	O
or	NN	O	O
abolishes	NN	O	O
in	NN	O	O
vitro	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
wild-type	NN	O	B-protein
proteins	NN	O	I-protein
of	NN	O	O
the	NN	O	O
same	NN	O	O
family	NN	O	O
(	NN	O	O
KBF1/p50	NN	O	B-protein
,	NN	O	O
c-	NN	O	B-protein
and	NN	O	I-protein
v-rel	NN	O	I-protein
)	NN	O	O
.	NN	O	O

This	NN	O	O
mutant	NN	O	O
also	NN	O	O
functions	NN	O	O
in	NN	O	O
vivo	NN	O	O
as	NN	O	O
a	NN	O	O
trans-acting	NN	O	B-protein
dominant	NN	O	I-protein
negative	NN	O	I-protein
regulator	NN	O	I-protein
:	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
inducibility	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
which	NN	O	O
contains	NN	O	O
two	NN	O	O
potential	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
sites	NN	O	O
)	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
is	NN	O	O
inhibited	NN	O	O
when	NN	O	O
it	NN	O	O
is	NN	O	O
co-transfected	NN	O	O
into	NN	O	O
CD4+	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
the	NN	O	O
delta	NN	O	B-protein
SP	NN	O	I-protein
mutant	NN	O	O
.	NN	O	O

Similarly	NN	O	O
the	NN	O	O
basal	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
TNF	NN	O	B-protein
or	NN	O	O
IL1-induced	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
I	NN	O	I-DNA
H-2Kb	NN	O	I-DNA
promoter	NN	O	I-DNA
can	NN	O	O
be	NN	O	O
inhibited	NN	O	O
by	NN	O	O
this	NN	O	O
mutant	NN	O	O
in	NN	O	O
two	NN	O	O
different	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
constitute	NN	O	O
the	NN	O	O
first	NN	O	O
formal	NN	O	O
demonstration	NN	O	O
that	NN	O	O
these	NN	O	O
genes	NN	O	O
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
rel/NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
platelet	NN	O	B-DNA
GPIIb	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	O
objectives	NN	O	O
in	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
thrombogenesis	NN	O	O
is	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
a	NN	O	O
hematopoietic	NN	O	B-cell_type
progenitor	NN	O	I-cell_type
is	NN	O	O
activated	NN	O	O
and	NN	O	O
committed	NN	O	O
to	NN	O	O
the	NN	O	O
megakaryocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

Recent	NN	O	O
development	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_line
cultures	NN	O	I-cell_line
of	NN	O	O
human	NN	O	B-cell_line
megakaryocytes	NN	O	I-cell_line
and	NN	O	O
the	NN	O	O
molecular	NN	O	O
cloning	NN	O	O
of	NN	O	O
genes	NN	O	O
that	NN	O	O
are	NN	O	O
specific	NN	O	O
to	NN	O	O
this	NN	O	O
lineage	NN	O	O
offer	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
getting	NN	O	O
some	NN	O	O
insights	NN	O	O
into	NN	O	O
the	NN	O	O
genetic	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
control	NN	O	O
megakaryocytopoiesis	NN	O	O
.	NN	O	O

One	NN	O	O
gene	NN	O	O
of	NN	O	O
interest	NN	O	O
is	NN	O	O
the	NN	O	O
glycoprotein	NN	O	B-DNA
IIb	NN	O	I-DNA
(	NN	O	I-DNA
GPIIb	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
;	NN	O	O
GPIIb	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
alpha	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
platelet	NN	O	B-protein
cytoadhesin	NN	O	I-protein
GPIIb-IIIa	NN	O	I-protein
,	NN	O	O
is	NN	O	O
produced	NN	O	O
in	NN	O	O
megakaryocytes	NN	O	B-cell_line
at	NN	O	O
an	NN	O	O
early	NN	O	O
stage	NN	O	O
of	NN	O	O
the	NN	O	O
differentiation	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
other	NN	O	O
subunit	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
,	NN	O	O
GPIIIa	NN	O	B-protein
,	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
other	NN	O	O
cells	NN	O	O
.	NN	O	O

For	NN	O	O
these	NN	O	O
reasons	NN	O	O
,	NN	O	O
the	NN	O	O
5'-flanking	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
GPIIb	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
regions	NN	O	O
that	NN	O	O
interact	NN	O	O
with	NN	O	O
DNA-binding	NN	O	B-protein
nuclear	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

A	NN	O	O
fragment	NN	O	O
extending	NN	O	O
from	NN	O	O
-643	NN	O	B-DNA
to	NN	O	I-DNA
+33	NN	O	I-DNA
is	NN	O	O
capable	NN	O	O
of	NN	O	O
controlling	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CAT	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
transfection	NN	O	O
experiments	NN	O	O
.	NN	O	O

Within	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
identified	NN	O	O
several	NN	O	O
sequences	NN	O	O
that	NN	O	O
are	NN	O	O
implicated	NN	O	O
in	NN	O	O
DNA	NN	O	O
protein	NN	O	O
interactions	NN	O	O
as	NN	O	O
shown	NN	O	O
in	NN	O	O
DNAse	NN	O	B-protein
I	NN	O	I-protein
footprints	NN	O	O
and	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

One	NN	O	O
region	NN	O	O
,	NN	O	O
centered	NN	O	O
at	NN	O	O
-54	NN	O	O
,	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
a	NN	O	O
nuclear	NN	O	B-DNA
factor	NN	O	I-DNA
E1-binding	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
a	NN	O	O
region	NN	O	O
located	NN	O	O
at	NN	O	O
position	NN	O	O
-233	NN	O	O
contains	NN	O	O
a	NN	O	O
CCAAT	NN	O	B-DNA
motif	NN	O	I-DNA
.	NN	O	O

Two	NN	O	O
domains	NN	O	O
centered	NN	O	O
at	NN	O	O
positions	NN	O	O
-345	NN	O	O
and	NN	O	O
-540	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
bind	NN	O	O
proteins	NN	O	O
that	NN	O	O
are	NN	O	O
present	NN	O	O
in	NN	O	O
megakaryocytic	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
nonrelated	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
two	NN	O	O
other	NN	O	O
domains	NN	O	O
,	NN	O	O
located	NN	O	O
at	NN	O	O
positions	NN	O	O
-460	NN	O	O
and	NN	O	O
-510	NN	O	O
,	NN	O	O
interact	NN	O	O
with	NN	O	O
proteins	NN	O	O
that	NN	O	O
are	NN	O	O
only	NN	O	O
present	NN	O	O
in	NN	O	O
megakaryocytic	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
region	NN	O	O
containing	NN	O	O
these	NN	O	O
two	NN	O	O
domains	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
activity	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
very	NN	O	O
likely	NN	O	O
that	NN	O	O
these	NN	O	O
domains	NN	O	O
bind	NN	O	O
megakaryocyte-specific	NN	O	B-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
acting	NN	O	O
as	NN	O	O
positive	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Lymphocyte	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
during	NN	O	O
depression	NN	O	O
and	NN	O	O
after	NN	O	O
clinical	NN	O	O
recovery	NN	O	O
.	NN	O	O

Lymphocyte	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
binding	NN	O	O
parameters	NN	O	O
were	NN	O	O
studied	NN	O	O
in	NN	O	O
15	NN	O	O
severely	NN	O	O
depressed	NN	O	O
patients	NN	O	O
during	NN	O	O
depression	NN	O	O
and	NN	O	O
after	NN	O	O
clinical	NN	O	O
recovery	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
15	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
difference	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
number	NN	O	O
or	NN	O	O
affinity	NN	O	O
between	NN	O	O
depressed	NN	O	O
patients	NN	O	O
and	NN	O	O
recovered	NN	O	O
or	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

Afternoon	NN	O	O
ACTH	NN	O	O
and	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
significantly	NN	O	O
between	NN	O	O
the	NN	O	O
three	NN	O	O
groups	NN	O	O
.	NN	O	O

No	NN	O	O
relationship	NN	O	O
could	NN	O	O
be	NN	O	O
established	NN	O	O
between	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
and	NN	O	O
antidepressant	NN	O	O
medication	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
view	NN	O	O
of	NN	O	O
an	NN	O	O
impaired	NN	O	O
ligand-induced	NN	O	O
plasticity	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
regulation	NN	O	O
rather	NN	O	O
than	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
of	NN	O	O
decreased	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
numbers	NN	O	O
during	NN	O	O
depression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Every	NN	O	O
enhancer	NN	O	O
works	NN	O	O
with	NN	O	O
every	NN	O	O
promoter	NN	O	B-DNA
for	NN	O	O
all	NN	O	O
the	NN	O	O
combinations	NN	O	O
tested	NN	O	O
:	NN	O	O
could	NN	O	O
new	NN	O	O
regulatory	NN	O	O
pathways	NN	O	O
evolve	NN	O	O
by	NN	O	O
enhancer	NN	O	O
shuffling	NN	O	O
?	NN	O	O

The	NN	O	O
promoters	NN	O	B-DNA
and	NN	O	O
enhancers	NN	O	B-DNA
of	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
genes	NN	O	O
are	NN	O	O
often	NN	O	O
conserved	NN	O	O
in	NN	O	O
evolution	NN	O	O
,	NN	O	O
and	NN	O	O
hence	NN	O	O
one	NN	O	O
might	NN	O	O
expect	NN	O	O
that	NN	O	O
a	NN	O	O
given	NN	O	O
enhancer	NN	O	B-DNA
has	NN	O	O
evolved	NN	O	O
to	NN	O	O
work	NN	O	O
best	NN	O	O
with	NN	O	O
its	NN	O	O
own	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

While	NN	O	O
this	NN	O	O
expectation	NN	O	O
may	NN	O	O
be	NN	O	O
realized	NN	O	O
in	NN	O	O
some	NN	O	O
cases	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
not	NN	O	O
found	NN	O	O
evidence	NN	O	O
for	NN	O	O
it	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
27	NN	O	O
combinations	NN	O	O
of	NN	O	O
different	NN	O	O
promoters	NN	O	B-DNA
and	NN	O	O
enhancers	NN	O	B-DNA
were	NN	O	O
tested	NN	O	O
by	NN	O	O
transfection	NN	O	O
into	NN	O	O
cultured	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
relative	NN	O	O
efficiency	NN	O	O
of	NN	O	O
the	NN	O	O
enhancers	NN	O	B-DNA
is	NN	O	O
approximately	NN	O	O
the	NN	O	O
same	NN	O	O
,	NN	O	O
irrespective	NN	O	O
of	NN	O	O
the	NN	O	O
type	NN	O	O
of	NN	O	O
promoter	NN	O	B-DNA
used	NN	O	O
,	NN	O	O
i.e.	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
strong	NN	O	O
preference	NN	O	O
for	NN	O	O
any	NN	O	O
given	NN	O	O
enhancer/	NN	O	O
promoter	NN	O	B-DNA
combination	NN	O	O
.	NN	O	O

Notably	NN	O	O
,	NN	O	O
we	NN	O	O
do	NN	O	O
not	NN	O	O
see	NN	O	O
particularly	NN	O	O
strong	NN	O	O
transcription	NN	O	O
when	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
kappa	NN	O	I-DNA
enhancer	NN	O	I-DNA
(	NN	O	O
or	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
)	NN	O	O
is	NN	O	O
used	NN	O	O
to	NN	O	O
activate	NN	O	O
a	NN	O	O
kappa	NN	O	O
gene	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
a	NN	O	O
generally	NN	O	O
permissive	NN	O	O
enhancer/	NN	O	O
promoter	NN	O	B-DNA
interaction	NN	O	O
is	NN	O	O
of	NN	O	O
evolutionary	NN	O	O
benefit	NN	O	O
for	NN	O	O
higher	NN	O	O
eukaryotes	NN	O	O
:	NN	O	O
by	NN	O	O
enhancer	NN	O	O
shuffling	NN	O	O
,	NN	O	O
genes	NN	O	O
could	NN	O	O
be	NN	O	O
easily	NN	O	O
brought	NN	O	O
under	NN	O	O
a	NN	O	O
new	NN	O	O
type	NN	O	O
of	NN	O	O
inducibility/cell	NN	O	O
type	NN	O	O
specificity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Towards	NN	O	O
a	NN	O	O
molecular	NN	O	O
understanding	NN	O	O
of	NN	O	O
T-cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Lymphoid	NN	O	O
differentiation	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
best	NN	O	O
studied	NN	O	O
examples	NN	O	O
of	NN	O	O
mammalian	NN	O	O
development	NN	O	O
.	NN	O	O

Here	NN	O	O
Hans	NN	O	O
Clevers	NN	O	O
and	NN	O	O
Michael	NN	O	O
Owen	NN	O	O
describe	NN	O	O
how	NN	O	O
the	NN	O	O
cloning	NN	O	O
of	NN	O	O
the	NN	O	O
genes	NN	O	O
that	NN	O	O
encode	NN	O	O
T-cell-specific	NN	O	B-protein
membrane	NN	O	I-protein
proteins	NN	O	I-protein
allows	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
control	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
T-cell	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Such	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
play	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
the	NN	O	O
mature	NN	O	O
T-cell	NN	O	O
phenotype	NN	O	O
by	NN	O	O
functioning	NN	O	O
as	NN	O	O
'master	NN	O	O
regulators	NN	O	O
of	NN	O	O
T-cell	NN	O	O
differentiation'	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
study	NN	O	O
on	NN	O	O
the	NN	O	O
circadian	NN	O	O
rhythm	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Circadian	NN	O	O
rhythm	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
was	NN	O	O
studied	NN	O	O
in	NN	O	O
the	NN	O	O
rat	NN	O	B-cell_type
liver	NN	O	I-cell_type
and	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

For	NN	O	O
rats	NN	O	O
exposed	NN	O	O
to	NN	O	O
a	NN	O	O
natural	NN	O	O
environmental	NN	O	O
photic	NN	O	O
cycle	NN	O	O
or	NN	O	O
a	NN	O	O
12L	NN	O	O
:	NN	O	O
12D	NN	O	O
artificial	NN	O	O
light	NN	O	O
regime	NN	O	O
,	NN	O	O
peak	NN	O	O
values	NN	O	O
of	NN	O	O
hepatic	NN	O	B-protein
GR	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
between	NN	O	O
23	NN	O	O
:	NN	O	O
00	NN	O	O
and	NN	O	O
02	NN	O	O
:	NN	O	O
00	NN	O	O
h	NN	O	O
.	NN	O	O

Except	NN	O	O
for	NN	O	O
a	NN	O	O
4-hour	NN	O	O
advancement	NN	O	O
of	NN	O	O
the	NN	O	O
peak	NN	O	O
,	NN	O	O
a	NN	O	O
similar	NN	O	O
circadian	NN	O	O
rhythm	NN	O	O
of	NN	O	O
hepatic	NN	O	B-protein
GR	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
in	NN	O	O
rats	NN	O	O
reared	NN	O	O
under	NN	O	O
a	NN	O	O
reversed	NN	O	O
lighting	NN	O	O
regimen	NN	O	O
(	NN	O	O
12D	NN	O	O
:	NN	O	O
12L	NN	O	O
;	NN	O	O
lights	NN	O	O
on	NN	O	O
between	NN	O	O
18	NN	O	O
:	NN	O	O
30	NN	O	O
and	NN	O	O
06	NN	O	O
:	NN	O	O
30	NN	O	O
h	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
human	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
peak	NN	O	O
value	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
parallel	NN	O	O
that	NN	O	O
of	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
with	NN	O	O
high	NN	O	O
and	NN	O	O
low	NN	O	O
values	NN	O	O
detected	NN	O	O
at	NN	O	O
04	NN	O	O
:	NN	O	O
00-08	NN	O	O
:	NN	O	O
00	NN	O	O
h	NN	O	O
and	NN	O	O
23	NN	O	O
:	NN	O	O
00-24	NN	O	O
:	NN	O	O
00	NN	O	O
h	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

In	NN	O	O
patients	NN	O	O
suffering	NN	O	O
from	NN	O	O
Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
,	NN	O	O
the	NN	O	O
circadian	NN	O	O
rhythm	NN	O	O
of	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
either	NN	O	O
disappeared	NN	O	O
or	NN	O	O
was	NN	O	O
inverted	NN	O	O
while	NN	O	O
that	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
did	NN	O	O
not	NN	O	O
significantly	NN	O	O
deviate	NN	O	O
from	NN	O	O
the	NN	O	O
normal	NN	O	O
subjects	NN	O	O
.	NN	O	O

For	NN	O	O
apoplexic	NN	O	O
patients	NN	O	O
with	NN	O	O
lesions	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
base	NN	O	O
of	NN	O	O
the	NN	O	O
brain	NN	O	O
as	NN	O	O
indicated	NN	O	O
by	NN	O	O
computerized	NN	O	O
tomography	NN	O	O
,	NN	O	O
the	NN	O	O
diurnal	NN	O	O
variation	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
was	NN	O	O
abolished	NN	O	O
.	NN	O	O

Conversely	NN	O	O
,	NN	O	O
diurnal	NN	O	O
rhythmicity	NN	O	O
persisted	NN	O	O
in	NN	O	O
apoplexy	NN	O	O
patients	NN	O	O
whose	NN	O	O
lesions	NN	O	O
were	NN	O	O
in	NN	O	O
the	NN	O	O
cerebral	NN	O	O
cortex	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
we	NN	O	O
postulated	NN	O	O
that	NN	O	O
the	NN	O	O
circadian	NN	O	O
modification	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
was	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
diurnal	NN	O	O
fluctuations	NN	O	O
in	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
level	NN	O	O
or	NN	O	O
the	NN	O	O
circadian	NN	O	O
variations	NN	O	O
in	NN	O	O
environmental	NN	O	O
lighting	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
rhythmicity	NN	O	O
might	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
'circadian	NN	O	O
pacemaker	NN	O	O
'	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	O
basal	NN	O	O
brain	NN	O	O
.	NN	O	O

These	NN	O	O
diurnal	NN	O	O
variations	NN	O	O
in	NN	O	O
GR	NN	O	B-protein
might	NN	O	O
serve	NN	O	O
to	NN	O	O
coordinate	NN	O	O
the	NN	O	O
reactivity	NN	O	O
of	NN	O	O
the	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
cortisol	NN	O	O
because	NN	O	O
the	NN	O	O
diurnal	NN	O	O
rhythms	NN	O	O
of	NN	O	O
a	NN	O	O
GR	NN	O	B-protein
-mediated	NN	O	O
response	NN	O	O
,	NN	O	O
the	NN	O	O
fractional	NN	O	O
inhibition	NN	O	O
of	NN	O	O
chemotactic	NN	O	O
migration	NN	O	O
rate	NN	O	O
of	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
by	NN	O	O
cortisol	NN	O	O
,	NN	O	O
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
synchronous	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
requirements	NN	O	O
for	NN	O	O
U2	NN	O	B-RNA
snRNA	NN	O	I-RNA
-encoding	NN	O	O
gene	NN	O	O
activation	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Transcription	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	O
U2	NN	O	B-DNA
small	NN	O	I-DNA
nuclear	NN	O	I-DNA
RNA	NN	O	I-DNA
(	NN	O	I-DNA
snRNA	NN	O	I-DNA
)	NN	O	I-DNA
-encoding	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
requires	NN	O	O
a	NN	O	O
distal	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
composed	NN	O	O
of	NN	O	O
one	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
(	NN	O	O
Oct	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
three	NN	O	O
Sp	NN	O	B-DNA
1-binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
study	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
requirement	NN	O	O
in	NN	O	O
B-cells	NN	O	B-cell_type
,	NN	O	O
different	NN	O	O
U2	NN	O	B-DNA
enhancer	NN	O	I-DNA
constructions	NN	O	I-DNA
were	NN	O	O
transfected	NN	O	O
into	NN	O	O
the	NN	O	O
lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
BJA-B	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
U2	NN	O	O
snRNA	NN	O	O
transcription	NN	O	O
in	NN	O	O
B-cells	NN	O	B-cell_type
also	NN	O	O
requires	NN	O	O
an	NN	O	O
enhancer	NN	O	O
comprising	NN	O	O
both	NN	O	O
the	NN	O	O
Oct	NN	O	B-DNA
and	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
Sp	NN	O	B-DNA
1-binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
all	NN	O	O
the	NN	O	O
Sp	NN	O	B-DNA
1-binding	NN	O	I-DNA
sites	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
reduces	NN	O	O
transcription	NN	O	O
by	NN	O	O
80-90	NN	O	O
%	NN	O	O
in	NN	O	O
HeLa	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
BJA-B	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
removal	NN	O	O
of	NN	O	O
the	NN	O	O
octamer-binding	NN	O	B-DNA
site	NN	O	I-DNA
reduces	NN	O	O
transcription	NN	O	O
to	NN	O	O
levels	NN	O	O
below	NN	O	O
detection	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Enhancers	NN	O	O
containing	NN	O	O
a	NN	O	O
single	NN	O	O
Oct	NN	O	B-DNA
have	NN	O	O
,	NN	O	O
nevertheless	NN	O	O
,	NN	O	O
the	NN	O	O
capacity	NN	O	O
to	NN	O	O
partially	NN	O	O
activate	NN	O	O
U2	NN	O	B-RNA
snRNA	NN	O	I-RNA
transcription	NN	O	O
in	NN	O	O
both	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
which	NN	O	O
only	NN	O	O
OTF-1	NN	O	O
is	NN	O	O
expressed	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
BJA-B	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
which	NN	O	O
OTF-2	NN	O	B-protein
is	NN	O	O
the	NN	O	O
predominantly	NN	O	O
expressed	NN	O	O
octamer-binding	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
most	NN	O	O
likely	NN	O	O
interpretation	NN	O	O
of	NN	O	O
our	NN	O	O
results	NN	O	O
is	NN	O	O
that	NN	O	O
both	NN	O	O
the	NN	O	O
ubiquitous	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
OTF-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
B-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
OTF-2	NN	O	B-protein
,	NN	O	O
can	NN	O	O
activate	NN	O	O
U2	NN	O	B-RNA
snRNA	NN	O	I-RNA
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
also	NN	O	O
revealed	NN	O	O
a	NN	O	O
similar	NN	O	O
functional	NN	O	O
cooperation	NN	O	O
between	NN	O	O
the	NN	O	O
transcription	NN	O	O
factors	NN	O	O
which	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
Oct	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
adjacent	NN	O	O
Sp	NN	O	B-DNA
1-binding	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
BJA-B	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
as	NN	O	O
has	NN	O	O
been	NN	O	O
observed	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
since	NN	O	O
a	NN	O	O
template	NN	O	O
which	NN	O	O
contains	NN	O	O
a	NN	O	O
weak	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
OTFs	NN	O	B-DNA
expresses	NN	O	O
wild-type	NN	O	O
levels	NN	O	O
of	NN	O	O
U2	NN	O	B-RNA
snRNA	NN	O	I-RNA
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
when	NN	O	O
the	NN	O	O
weak	NN	O	O
octamer-binding	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
combined	NN	O	O
with	NN	O	O
a	NN	O	O
Sp	NN	O	B-DNA
1-binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

One	NN	O	O
base	NN	O	O
pair	NN	O	O
change	NN	O	O
abolishes	NN	O	O
the	NN	O	O
T	NN	O	O
cell-restricted	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
kB-like	NN	O	B-DNA
proto-enhancer	NN	O	I-DNA
element	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
inducible	NN	O	O
,	NN	O	O
T	NN	O	B-DNA
cell-specific	NN	O	I-DNA
enhancers	NN	O	I-DNA
of	NN	O	O
murine	NN	O	B-DNA
and	NN	O	I-DNA
human	NN	O	I-DNA
Interleukin	NN	O	I-DNA
2	NN	O	I-DNA
(	NN	O	I-DNA
Il-2	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
contain	NN	O	O
the	NN	O	O
kB-like	NN	O	B-DNA
sequence	NN	O	I-DNA
GGGATTTCACC	NN	O	O
as	NN	O	O
an	NN	O	O
essential	NN	O	B-DNA
cis-acting	NN	O	I-DNA
enhancer	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

When	NN	O	O
cloned	NN	O	O
in	NN	O	O
multiple	NN	O	O
copies	NN	O	O
this	NN	O	O
so-called	NN	O	O
TCEd	NN	O	B-DNA
(	NN	O	O
distal	NN	O	B-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
element	NN	O	I-DNA
)	NN	O	O
acts	NN	O	O
as	NN	O	O
an	NN	O	O
inducible	NN	O	O
proto-enhancer	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
E14	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphoma	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
extracts	NN	O	O
of	NN	O	O
induced	NN	O	O
,	NN	O	O
Il-2	NN	O	B-cell_line
secreting	NN	O	I-cell_line
El4	NN	O	I-cell_line
cells	NN	O	I-cell_line
three	NN	O	O
individual	NN	O	O
protein	NN	O	B-protein
factors	NN	O	I-protein
bind	NN	O	O
to	NN	O	O
TCEd	NN	O	B-DNA
DNA	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
prominent	NN	O	O
factor	NN	O	O
,	NN	O	O
named	NN	O	O
TCF-1	NN	O	B-protein
(	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
)	NN	O	O
,	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
proto-enhancer	NN	O	O
activity	NN	O	O
of	NN	O	O
TCEd	NN	O	O
.	NN	O	O

TCF-1	NN	O	B-protein
consists	NN	O	O
of	NN	O	O
two	NN	O	O
polypeptides	NN	O	O
of	NN	O	O
about	NN	O	O
50	NN	O	O
kD	NN	O	O
and	NN	O	O
105	NN	O	O
kD	NN	O	O
;	NN	O	O
the	NN	O	O
former	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
50	NN	O	B-protein
kD	NN	O	I-protein
polypeptide	NN	O	I-protein
of	NN	O	O
NF-kB	NN	O	B-protein
.	NN	O	O

Purified	NN	O	B-protein
NF-kB	NN	O	I-protein
is	NN	O	O
also	NN	O	O
able	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
TCEd	NN	O	B-DNA
,	NN	O	O
but	NN	O	O
TCF-1	NN	O	B-protein
binds	NN	O	O
stronger	NN	O	O
than	NN	O	O
NF-kB	NN	O	B-protein
to	NN	O	O
TCEd	NN	O	B-DNA
DNA	NN	O	O
.	NN	O	O

The	NN	O	O
conversion	NN	O	O
of	NN	O	O
the	NN	O	O
TCEd	NN	O	B-DNA
to	NN	O	O
a	NN	O	O
'perfect	NN	O	B-DNA
'	NN	O	I-DNA
NF-kB	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
leads	NN	O	O
to	NN	O	O
a	NN	O	O
tighter	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kB	NN	O	B-protein
to	NN	O	O
TCEd	NN	O	B-DNA
DNA	NN	O	I-DNA
and	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
functional	NN	O	O
consequence	NN	O	O
,	NN	O	O
to	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
'converted	NN	O	B-DNA
'	NN	O	I-DNA
TCEd	NN	O	I-DNA
motifs	NN	O	I-DNA
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
substitution	NN	O	O
of	NN	O	O
the	NN	O	O
underlined	NN	O	O
A	NN	O	O
residue	NN	O	O
to	NN	O	O
a	NN	O	O
C	NN	O	O
within	NN	O	O
the	NN	O	O
GGGATTTCACC	NN	O	B-DNA
motif	NN	O	I-DNA
abolishes	NN	O	O
its	NN	O	O
T	NN	O	O
cell-restricted	NN	O	O
activity	NN	O	O
and	NN	O	O
leads	NN	O	O
to	NN	O	O
its	NN	O	O
functioning	NN	O	O
in	NN	O	O
both	NN	O	O
El4	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
lymphocyte-specific	NN	O	O
factors	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
TCEd	NN	O	B-DNA
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
T	NN	O	O
cell	NN	O	O
specific-transcription	NN	O	O
of	NN	O	O
the	NN	O	O
Il-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
erythroid	NN	O	B-DNA
specific	NN	O	I-DNA
enhancer	NN	O	I-DNA
upstream	NN	O	O
to	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
cell-type	NN	O	B-protein
specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
GATA-1	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
hemopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
where	NN	O	O
it	NN	O	O
mediates	NN	O	O
the	NN	O	O
cell-type	NN	O	O
specific	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
genes	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
the	NN	O	O
mouse	NN	O	O
and	NN	O	O
human	NN	O	O
GATA-1	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
region	NN	O	O
upstream	NN	O	O
to	NN	O	O
the	NN	O	O
first	NN	O	O
exon	NN	O	O
,	NN	O	O
and	NN	O	O
highly	NN	O	O
conserved	NN	O	O
between	NN	O	O
mouse	NN	O	O
and	NN	O	O
man	NN	O	O
,	NN	O	O
acts	NN	O	O
as	NN	O	O
an	NN	O	O
erythroid	NN	O	O
specific	NN	O	O
enhancer	NN	O	O
in	NN	O	O
transient	NN	O	O
assays	NN	O	O
,	NN	O	O
if	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
GATA-1	NN	O	B-protein
or	NN	O	O
to	NN	O	O
the	NN	O	O
SV40	NN	O	O
promoter	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	O
is	NN	O	O
almost	NN	O	O
completely	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
integrity	NN	O	O
of	NN	O	O
a	NN	O	O
dimeric	NN	O	B-DNA
GATA-1	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Demonstration	NN	O	O
of	NN	O	O
a	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25-dihydroxyvitamin	NN	O	I-protein
D3-responsive	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
immunologic	NN	O	O
crossreactivity	NN	O	O
and	NN	O	O
inverse	NN	O	O
regulation	NN	O	O
with	NN	O	O
the	NN	O	O
vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
with	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
recognizing	NN	O	O
a	NN	O	O
17-amino	NN	O	B-protein
acid	NN	O	I-protein
epitope	NN	O	I-protein
of	NN	O	O
the	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25-dihydroxyvitamin	NN	O	I-protein
D3	NN	O	I-protein
[	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
]	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
we	NN	O	O
have	NN	O	O
detected	NN	O	O
two	NN	O	O
crossreacting	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
activated	NN	O	B-cell_type
normal	NN	O	I-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
smaller	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
proteins	NN	O	O
(	NN	O	O
50	NN	O	B-protein
kDa	NN	O	I-protein
)	NN	O	O
was	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
the	NN	O	O
classical	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
,	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
classical	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
was	NN	O	O
upregulated	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
fashion	NN	O	O
by	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
.	NN	O	O

The	NN	O	O
larger	NN	O	O
crossreacting	NN	O	O
protein	NN	O	O
exhibited	NN	O	O
an	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
of	NN	O	O
80	NN	O	O
kDa	NN	O	O
,	NN	O	O
was	NN	O	O
localized	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
cytosol	NN	O	O
,	NN	O	O
and	NN	O	O
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
specific	NN	O	O
for	NN	O	O
activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
since	NN	O	O
it	NN	O	O
was	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
several	NN	O	O
other	NN	O	O
human	NN	O	B-cell_type
cells	NN	O	I-cell_type
including	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

More	NN	O	O
strikingly	NN	O	O
,	NN	O	O
the	NN	O	O
80-kDa	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
downregulated	NN	O	O
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
fashion	NN	O	O
by	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
;	NN	O	O
this	NN	O	O
effect	NN	O	O
was	NN	O	O
independent	NN	O	O
of	NN	O	O
the	NN	O	O
mode	NN	O	O
of	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
and	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
metabolite	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
two	NN	O	O
potent	NN	O	O
immunosuppressive	NN	O	O
agents	NN	O	O
,	NN	O	O
glucocorticoids	NN	O	O
and	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
also	NN	O	O
inhibited	NN	O	O
the	NN	O	O
80-kDa	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
treatment	NN	O	O
,	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
and	NN	O	O
ketoconazole	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GcR	NN	O	B-protein
)	NN	O	O
were	NN	O	O
determined	NN	O	O
by	NN	O	O
a	NN	O	O
whole	NN	O	O
cell	NN	O	O
assay	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
mononulear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
hMNL	NN	O	B-cell_type
)	NN	O	O
from	NN	O	O
control	NN	O	O
subjects	NN	O	O
,	NN	O	O
patients	NN	O	O
receiving	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
for	NN	O	O
systemic	NN	O	O
diseases	NN	O	O
and	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
patients	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
ketoconazole	NN	O	O
therapy	NN	O	O
.	NN	O	O

Prolonged	NN	O	O
corticosteroid	NN	O	O
treatment	NN	O	O
resulted	NN	O	O
in	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
GcR	NN	O	B-protein
,	NN	O	O
while	NN	O	O
the	NN	O	O
mean	NN	O	O
level	NN	O	O
of	NN	O	O
GcR	NN	O	B-protein
in	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
was	NN	O	O
normal	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
group	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
receptor	NN	O	O
levels	NN	O	O
and	NN	O	O
morning	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
values	NN	O	O
showed	NN	O	O
a	NN	O	O
negative	NN	O	O
correlation	NN	O	O
,	NN	O	O
indicating	NN	O	O
a	NN	O	O
subtle	NN	O	O
down-regulatory	NN	O	O
effect	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
GcR	NN	O	B-protein
were	NN	O	O
unaltered	NN	O	O
after	NN	O	O
these	NN	O	O
patients	NN	O	O
received	NN	O	O
ketoconazole	NN	O	O
,	NN	O	O
in	NN	O	O
spite	NN	O	O
of	NN	O	O
a	NN	O	O
marked	NN	O	O
reduction	NN	O	O
in	NN	O	O
morning	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
and	NN	O	O
urinary	NN	O	O
free	NN	O	O
cortisol	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
observed	NN	O	O
that	NN	O	O
ketoconazole	NN	O	O
was	NN	O	O
a	NN	O	O
weak	NN	O	O
competitor	NN	O	O
of	NN	O	O
GcR	NN	O	B-protein
in	NN	O	O
intact	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
although	NN	O	O
it	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
dexamethasone	NN	O	O
binding	NN	O	O
in	NN	O	O
cytosolic	NN	O	O
preparations	NN	O	O
from	NN	O	O
rat	NN	O	O
tissues	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
suggested	NN	O	O
that	NN	O	O
GcR	NN	O	B-protein
in	NN	O	O
hMNL	NN	O	B-cell_type
are	NN	O	O
down-regulated	NN	O	O
by	NN	O	O
synthetic	NN	O	O
steroids	NN	O	O
given	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
but	NN	O	O
they	NN	O	O
showed	NN	O	O
very	NN	O	O
mild	NN	O	O
down-regulation	NN	O	O
in	NN	O	O
hypercortisolemic	NN	O	O
patients	NN	O	O
suffering	NN	O	O
from	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
did	NN	O	O
not	NN	O	O
observed	NN	O	O
either	NN	O	O
up-regulation	NN	O	O
or	NN	O	O
antagonism	NN	O	O
of	NN	O	O
GcR	NN	O	B-protein
by	NN	O	O
ketoconazole	NN	O	O
treatment	NN	O	O
,	NN	O	O
at	NN	O	O
the	NN	O	O
time	NN	O	O
that	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
were	NN	O	O
reduced	NN	O	O
.	NN	O	O

This	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
beneficial	NN	O	O
effects	NN	O	O
of	NN	O	O
ketoconazole	NN	O	O
in	NN	O	O
Cushing	NN	O	O
's	NN	O	O
disease	NN	O	O
are	NN	O	O
due	NN	O	O
to	NN	O	O
adrenal	NN	O	O
cortisol	NN	O	O
suppression	NN	O	O
and	NN	O	O
not	NN	O	O
to	NN	O	O
interaction	NN	O	O
with	NN	O	O
GcR	NN	O	B-protein
of	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
GcR	NN	O	B-protein
regulation	NN	O	O
in	NN	O	O
hMNL	NN	O	B-cell_type
is	NN	O	O
a	NN	O	O
complex	NN	O	O
phenomenon	NN	O	O
awaiting	NN	O	O
further	NN	O	O
elucidation	NN	O	O
.	NN	O	O

-DOCSTART-	O

HTLV-1	NN	O	B-protein
Tax	NN	O	I-protein
induces	NN	O	O
expression	NN	O	O
of	NN	O	O
various	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
serum	NN	O	I-DNA
responsive	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HTLV-1	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
etiological	NN	O	O
agent	NN	O	O
of	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
(	NN	O	O
ATL	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
showed	NN	O	O
here	NN	O	O
by	NN	O	O
mobility-shift	NN	O	O
assay	NN	O	O
that	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
transformed	NN	O	O
with	NN	O	O
the	NN	O	O
virus	NN	O	O
contained	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activities	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
this	NN	O	O
result	NN	O	O
,	NN	O	O
these	NN	O	O
cell	NN	O	O
lines	NN	O	O
expressed	NN	O	O
increased	NN	O	O
levels	NN	O	O
of	NN	O	O
mRNAs	NN	O	B-RNA
encoding	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
Fra-1	NN	O	B-protein
,	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
JunB	NN	O	B-protein
,	NN	O	O
and	NN	O	O
JunD	NN	O	B-protein
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
gene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
transactivated	NN	O	O
by	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
Tax1	NN	O	B-protein
.	NN	O	O

By	NN	O	O
using	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
JPX-9	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Tax1	NN	O	B-protein
is	NN	O	O
inducible	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
mRNAs	NN	O	B-RNA
for	NN	O	O
Fra-1	NN	O	B-protein
,	NN	O	O
c-Jun	NN	O	B-protein
,	NN	O	O
and	NN	O	O
JunD	NN	O	B-protein
was	NN	O	O
also	NN	O	O
transactivated	NN	O	O
by	NN	O	O
Tax1	NN	O	B-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
Tax1	NN	O	B-protein
activated	NN	O	O
expression	NN	O	O
of	NN	O	O
two	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
having	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
motifs	NN	O	I-protein
,	NN	O	O
Egr-1	NN	O	B-protein
and	NN	O	O
Egr-2	NN	O	B-protein
,	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
cells	NN	O	O
.	NN	O	O

The	NN	O	O
Tax1-inducible	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
identified	NN	O	O
here	NN	O	O
are	NN	O	O
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
members	NN	O	O
of	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
genes	NN	O	I-DNA
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
growth	NN	O	O
signals	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
Tax1	NN	O	B-protein
was	NN	O	O
suggested	NN	O	O
to	NN	O	O
replace	NN	O	O
growth	NN	O	O
signals	NN	O	O
,	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
by	NN	O	O
this	NN	O	O
mechanism	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
characteristics	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
corticosteroid-resistant	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
corticosteroid	NN	O	O
resistance	NN	O	O
in	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
has	NN	O	O
been	NN	O	O
studied	NN	O	O
by	NN	O	O
determining	NN	O	O
the	NN	O	O
rank	NN	O	O
order	NN	O	O
of	NN	O	O
potency	NN	O	O
for	NN	O	O
different	NN	O	O
corticosteroids	NN	O	O
in	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
a	NN	O	O
3	NN	O	O
kD	NN	O	O
molecule	NN	O	O
from	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
isolated	NN	O	O
from	NN	O	O
corticosteroid-sensitive	NN	O	O
(	NN	O	O
CS	NN	O	O
)	NN	O	O
and	NN	O	O
corticosteroid-resistant	NN	O	O
(	NN	O	O
CR	NN	O	O
)	NN	O	O
asthmatic	NN	O	O
subjects	NN	O	O
,	NN	O	O
which	NN	O	O
augments	NN	O	O
leukotriene	NN	O	O
B4	NN	O	O
(	NN	O	O
LTB4	NN	O	B-protein
)	NN	O	O
generation	NN	O	O
by	NN	O	O
human	NN	O	B-cell_type
neutrophils	NN	O	I-cell_type
(	NN	O	O
PMN	NN	O	B-cell_type
)	NN	O	O
stimulated	NN	O	O
by	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
binding	NN	O	O
studies	NN	O	O
with	NN	O	O
(	NN	O	O
3H	NN	O	O
)	NN	O	O
dexamethasone	NN	O	O
have	NN	O	O
been	NN	O	O
performed	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
and	NN	O	O
receptor	NN	O	O
numbers	NN	O	O
(	NN	O	O
Ro	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
monocytes	NN	O	B-cell_type
of	NN	O	O
these	NN	O	O
two	NN	O	O
groups	NN	O	O
of	NN	O	O
subjects	NN	O	O
.	NN	O	O

The	NN	O	O
concentration	NN	O	O
of	NN	O	O
corticosteroid	NN	O	O
producing	NN	O	O
50	NN	O	O
%	NN	O	O
inhibition	NN	O	O
(	NN	O	O
IC50	NN	O	O
)	NN	O	O
was	NN	O	O
600	NN	O	O
nM	NN	O	O
,	NN	O	O
70	NN	O	O
nM	NN	O	O
,	NN	O	O
and	NN	O	O
0.5	NN	O	O
nM	NN	O	O
for	NN	O	O
hydrocortisone	NN	O	O
,	NN	O	O
methylprednisolone	NN	O	O
,	NN	O	O
and	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
CS	NN	O	O
individuals	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
only	NN	O	O
weak	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
enhancing	NN	O	O
activity	NN	O	O
by	NN	O	O
the	NN	O	O
corticosteroids	NN	O	O
in	NN	O	O
the	NN	O	O
CR	NN	O	O
asthmatic	NN	O	O
individuals	NN	O	O
.	NN	O	O

The	NN	O	O
dexamethasone	NN	O	O
Kd	NN	O	O
was	NN	O	O
2.45	NN	O	O
+/-	NN	O	O
0.58	NN	O	O
nM	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
6	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
CS	NN	O	O
group	NN	O	O
and	NN	O	O
1.6	NN	O	O
+/-	NN	O	O
0.35	NN	O	O
nM	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
6	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
CR	NN	O	O
group	NN	O	O
of	NN	O	O
patients	NN	O	O
(	NN	O	O
p	NN	O	O
=	NN	O	O
0.14	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
Ro	NN	O	O
in	NN	O	O
the	NN	O	O
CS	NN	O	O
group	NN	O	O
was	NN	O	O
3	NN	O	O
,	NN	O	O
605	NN	O	O
+/-	NN	O	O
984	NN	O	O
binding	NN	O	O
sites	NN	O	O
per	NN	O	O
nucleus	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
6	NN	O	O
)	NN	O	O
and	NN	O	O
4	NN	O	O
,	NN	O	O
757	NN	O	O
+/-	NN	O	O
692	NN	O	O
binding	NN	O	O
sites	NN	O	O
per	NN	O	O
nucleus	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
6	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
CR	NN	O	O
group	NN	O	O
(	NN	O	O
p	NN	O	O
=	NN	O	O
0.23	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
corticosteroid	NN	O	O
resistance	NN	O	O
in	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
corticosteroid	NN	O	O
receptor	NN	O	O
characteristics	NN	O	O
.	NN	O	O

-DOCSTART-	O

Vitamin	NN	O	B-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Signal	NN	O	O
requirements	NN	O	O
and	NN	O	O
characterization	NN	O	O
by	NN	O	O
western	NN	O	O
blots	NN	O	O
and	NN	O	O
DNA	NN	O	O
sequencing	NN	O	O
.	NN	O	O

The	NN	O	O
signals	NN	O	O
controlling	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
protein	NN	O	I-protein
for	NN	O	O
1	NN	O	O
alpha	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
in	NN	O	O
normal	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
relationship	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
to	NN	O	O
the	NN	O	O
classical	NN	O	B-protein
vitamin	NN	O	I-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
were	NN	O	O
examined	NN	O	O
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
activated	NN	O	O
with	NN	O	O
the	NN	O	O
OKT3	NN	O	B-protein
antibody	NN	O	I-protein
to	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
expressed	NN	O	O
fewer	NN	O	O
binding	NN	O	O
sites	NN	O	O
as	NN	O	O
compared	NN	O	O
to	NN	O	O
lymphocytes	NN	O	B-cell_type
that	NN	O	O
were	NN	O	O
activated	NN	O	O
by	NN	O	O
the	NN	O	O
polyclonal	NN	O	O
activator	NN	O	O
phytohemagglutinin	NN	O	B-protein
(	NN	O	O
PHA	NN	O	B-protein
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
combination	NN	O	O
of	NN	O	O
OKT3	NN	O	B-protein
and	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
produced	NN	O	O
a	NN	O	O
concentration	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
PHA-activated	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
receptor	NN	O	O
from	NN	O	O
OKT3	NN	O	B-protein
and	NN	O	O
OKT3	NN	O	B-protein
+	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
-activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
exhibited	NN	O	O
decreased	NN	O	O
binding	NN	O	O
to	NN	O	O
DNA-cellulose	NN	O	O
compared	NN	O	O
to	NN	O	O
PHA-activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
lymphocytes	NN	O	B-cell_type
activated	NN	O	O
either	NN	O	O
by	NN	O	O
PHA	NN	O	B-protein
or	NN	O	O
OKT3	NN	O	B-protein
(	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
,	NN	O	O
a	NN	O	O
50-kDa	NN	O	B-protein
species	NN	O	I-protein
cross-reacting	NN	O	O
with	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
against	NN	O	O
the	NN	O	O
intestinal	NN	O	B-protein
vitamin	NN	O	I-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
RNA	NN	O	O
from	NN	O	O
activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
was	NN	O	O
amplified	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
using	NN	O	O
oligonucleotide	NN	O	O
primers	NN	O	O
flanking	NN	O	O
the	NN	O	O
196	NN	O	B-DNA
base	NN	O	I-DNA
pair	NN	O	I-DNA
long	NN	O	I-DNA
region	NN	O	I-DNA
encoding	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
intestinal	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
amplified	NN	O	O
product	NN	O	O
showed	NN	O	O
an	NN	O	O
identical	NN	O	B-DNA
nucleotide	NN	O	I-DNA
sequence	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
intestinal	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25-	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptor	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
is	NN	O	O
triggered	NN	O	O
by	NN	O	O
distinct	NN	O	O
and	NN	O	O
contingent	NN	O	O
signals	NN	O	O
,	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
protein	NN	O	O
and	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
encoding	NN	O	O
it	NN	O	O
are	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
classical	NN	O	B-protein
vitamin	NN	O	I-protein
D	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Comparison	NN	O	O
of	NN	O	O
constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
transcriptional	NN	O	O
enhancement	NN	O	O
mediated	NN	O	O
by	NN	O	O
kappa	NN	O	B-DNA
B-related	NN	O	I-DNA
sequences	NN	O	I-DNA
:	NN	O	O
modulation	NN	O	O
of	NN	O	O
activity	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
human	NN	O	B-DNA
T-cell	NN	O	I-DNA
leukemia	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
I	NN	O	I-DNA
tax	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sequence	NN	O	I-DNA
(	NN	O	O
GGGACTTTCC	NN	O	O
)	NN	O	O
binds	NN	O	O
a	NN	O	O
factor	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
that	NN	O	O
is	NN	O	O
constitutively	NN	O	O
found	NN	O	O
in	NN	O	O
its	NN	O	O
functional	NN	O	O
,	NN	O	O
DNA	NN	O	O
binding	NN	O	O
form	NN	O	O
only	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
factor	NN	O	O
with	NN	O	O
apparently	NN	O	O
indistinguishable	NN	O	O
sequence	NN	O	O
specificity	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
many	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
,	NN	O	O
where	NN	O	O
it	NN	O	O
is	NN	O	O
used	NN	O	O
to	NN	O	O
regulate	NN	O	O
inducible	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

For	NN	O	O
example	NN	O	O
,	NN	O	O
kappa	NN	O	B-DNA
B-related	NN	O	I-DNA
sequences	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
few	NN	O	O
inducible	NN	O	O
genes	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
interleukin	NN	O	B-DNA
2	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha-chain	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
beta-interferon	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
these	NN	O	O
genes	NN	O	O
are	NN	O	O
not	NN	O	O
constitutively	NN	O	O
active	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
other	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
must	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
determining	NN	O	O
the	NN	O	O
patterns	NN	O	O
of	NN	O	O
expression	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
the	NN	O	O
constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
mediated	NN	O	O
by	NN	O	O
five	NN	O	O
kappa	NN	O	B-DNA
B-related	NN	O	I-DNA
sequence	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
two	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
in	NN	O	O
S194	NN	O	B-cell_line
plasma	NN	O	I-cell_line
cells	NN	O	I-cell_line
the	NN	O	O
activity	NN	O	O
of	NN	O	O
each	NN	O	O
element	NN	O	O
correlates	NN	O	O
well	NN	O	O
with	NN	O	O
the	NN	O	O
relative	NN	O	O
affinity	NN	O	O
of	NN	O	O
B-cell-derived	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
for	NN	O	O
that	NN	O	O
element	NN	O	O
.	NN	O	O

This	NN	O	O
leads	NN	O	O
to	NN	O	O
significantly	NN	O	O
lower	NN	O	O
transcription	NN	O	O
enhancement	NN	O	O
by	NN	O	O
sites	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
receptor	NN	O	I-protein
or	NN	O	O
T-cell	NN	O	B-DNA
receptor	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
S194	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
either	NN	O	O
EL-4	NN	O	B-cell_line
(	NN	O	I-cell_line
T	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
S194	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
both	NN	O	O
lower-affinity	NN	O	O
sites	NN	O	O
can	NN	O	O
be	NN	O	O
significantly	NN	O	O
induced	NN	O	O
by	NN	O	O
the	NN	O	O
tax	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
,	NN	O	O
showing	NN	O	O
that	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
can	NN	O	O
be	NN	O	O
modulated	NN	O	O
even	NN	O	O
in	NN	O	O
a	NN	O	O
B-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
that	NN	O	O
constitutively	NN	O	O
expresses	NN	O	O
this	NN	O	O
factor	NN	O	O
.	NN	O	O

-DOCSTART-	O

Specific	NN	O	O
depletion	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell	NN	O	B-cell_line
population	NN	O	I-cell_line
induced	NN	O	O
by	NN	O	O
aberrant	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
interferon	NN	O	I-DNA
regulatory	NN	O	I-DNA
factor	NN	O	I-DNA
1	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Interferons	NN	O	B-protein
(	NN	O	O
IFNs	NN	O	B-protein
)	NN	O	O
are	NN	O	O
well	NN	O	O
known	NN	O	O
both	NN	O	O
as	NN	O	O
antiviral	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
as	NN	O	O
potent	NN	O	O
regulators	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
IFNs	NN	O	B-protein
inhibit	NN	O	O
growth	NN	O	O
of	NN	O	O
various	NN	O	O
normal	NN	O	B-cell_type
and	NN	O	O
transformed	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
.	NN	O	O

Previously	NN	O	O
,	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
IRF-1	NN	O	B-protein
(	NN	O	O
interferon	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
type	NN	O	B-DNA
I	NN	O	I-DNA
IFN	NN	O	I-DNA
and	NN	O	O
some	NN	O	O
IFN-inducible	NN	O	B-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
identified	NN	O	O
and	NN	O	O
cloned	NN	O	O
.	NN	O	O

Since	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-protein
gene	NN	O	O
is	NN	O	O
both	NN	O	O
virus	NN	O	O
and	NN	O	O
IFN	NN	O	B-protein
inducible	NN	O	O
,	NN	O	O
an	NN	O	O
intriguing	NN	O	O
issue	NN	O	O
is	NN	O	O
raised	NN	O	O
as	NN	O	O
to	NN	O	O
whether	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
functioning	NN	O	O
in	NN	O	O
IFN	NN	O	B-protein
-mediated	NN	O	O
regulation	NN	O	O
of	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
generated	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
carrying	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IRF-1	NN	O	I-DNA
gene	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
heavy-chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
the	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
,	NN	O	O
all	NN	O	O
the	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
examined	NN	O	O
showed	NN	O	O
a	NN	O	O
dramatic	NN	O	O
reduction	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
(	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
)	NN	O	O
.	NN	O	O

Preparation	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
chimeric	NN	O	O
mice	NN	O	O
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
bone	NN	O	O
marrow	NN	O	O
is	NN	O	O
the	NN	O	O
effective	NN	O	O
site	NN	O	O
for	NN	O	O
specific	NN	O	O
depletion	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell	NN	O	B-cell_type
population	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
fact	NN	O	O
,	NN	O	O
transgenic	NN	O	B-cell_line
bone	NN	O	I-cell_line
marrow	NN	O	I-cell_line
cells	NN	O	I-cell_line
cocultured	NN	O	O
with	NN	O	O
a	NN	O	O
bone	NN	O	B-cell_line
marrow-derived	NN	O	I-cell_line
stromal	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
revealed	NN	O	O
an	NN	O	O
altered	NN	O	O
B-cell	NN	O	B-cell_type
maturation	NN	O	O
pattern	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	NN	O	O
of	NN	O	O
a	NN	O	O
cofactor	NN	O	B-protein
that	NN	O	O
regulates	NN	O	O
dimerization	NN	O	O
of	NN	O	O
a	NN	O	O
mammalian	NN	O	B-protein
homeodomain	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Dimerization	NN	O	O
among	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
has	NN	O	O
become	NN	O	O
a	NN	O	O
recurrent	NN	O	O
theme	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
eukaryotic	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Hepatocyte	NN	O	B-protein
nuclear	NN	O	I-protein
factor-1	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
HNF-1	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
homeodomain-containing	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
dimer	NN	O	B-protein
.	NN	O	O

A	NN	O	O
dimerization	NN	O	B-protein
cofactor	NN	O	I-protein
of	NN	O	O
HNF-1	NN	O	B-protein
alpha	NN	O	I-protein
(	NN	O	O
DCoH	NN	O	B-protein
)	NN	O	O
was	NN	O	O
identified	NN	O	O
that	NN	O	O
displayed	NN	O	O
a	NN	O	O
restricted	NN	O	O
tissue	NN	O	O
distribution	NN	O	O
and	NN	O	O
did	NN	O	O
not	NN	O	O
bind	NN	O	O
to	NN	O	O
DNA	NN	O	O
,	NN	O	O
but	NN	O	O
,	NN	O	O
rather	NN	O	O
,	NN	O	O
selectively	NN	O	O
stabilized	NN	O	O
HNF-1	NN	O	B-protein
alpha	NN	O	I-protein
dimers	NN	O	I-protein
.	NN	O	O

The	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
stable	NN	O	O
tetrameric	NN	O	B-protein
DCoH-HNF-1	NN	O	I-protein
alpha	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
which	NN	O	O
required	NN	O	O
the	NN	O	O
dimerization	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
HNF-1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
did	NN	O	O
not	NN	O	O
change	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
of	NN	O	O
HNF-1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
but	NN	O	O
enhanced	NN	O	O
its	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
DCoH	NN	O	B-protein
did	NN	O	O
not	NN	O	O
confer	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
to	NN	O	O
the	NN	O	O
GAL4	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
DCoH	NN	O	B-protein
regulates	NN	O	O
formation	NN	O	O
of	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
tetrameric	NN	O	B-protein
complexes	NN	O	I-protein
and	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
developmental	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
complex	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	O
resistance	NN	O	O
in	NN	O	O
chronic	NN	O	O
asthma	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	O
pharmacokinetics	NN	O	O
,	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
characteristics	NN	O	O
,	NN	O	O
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
proliferation	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

A	NN	O	O
total	NN	O	O
of	NN	O	O
37	NN	O	O
chronic	NN	O	O
,	NN	O	O
severe	NN	O	O
,	NN	O	O
nonsmoking	NN	O	O
asthmatic	NN	O	O
patients	NN	O	O
with	NN	O	O
documented	NN	O	O
reversible	NN	O	O
airways	NN	O	O
obstruction	NN	O	O
were	NN	O	O
classified	NN	O	O
as	NN	O	O
glucocorticoid-sensitive	NN	O	O
or	NN	O	O
-resistant	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
changes	NN	O	O
in	NN	O	O
FEV1	NN	O	O
,	NN	O	O
FVC	NN	O	O
,	NN	O	O
and	NN	O	O
peak	NN	O	O
expiratory	NN	O	O
flow	NN	O	O
(	NN	O	O
PEF	NN	O	O
)	NN	O	O
after	NN	O	O
oral	NN	O	O
prednisolone	NN	O	O
.	NN	O	O

The	NN	O	O
resistant	NN	O	O
patients	NN	O	O
showed	NN	O	O
no	NN	O	O
significant	NN	O	O
improvements	NN	O	O
in	NN	O	O
airflow	NN	O	O
limitation	NN	O	O
.	NN	O	O

Phytohemagglutinin	NN	O	B-protein
(	NN	O	O
PHA	NN	O	B-protein
)	NN	O	O
-induced	NN	O	O
proliferation	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
sensitive	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
resistant	NN	O	O
asthmatic	NN	O	O
patients	NN	O	O
was	NN	O	O
significantly	NN	O	O
(	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0.01	NN	O	O
)	NN	O	O
inhibited	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
mol/L	NN	O	O
)	NN	O	O
,	NN	O	O
reflecting	NN	O	O
a	NN	O	O
shift	NN	O	O
of	NN	O	O
the	NN	O	O
dose-response	NN	O	O
curve	NN	O	O
.	NN	O	O

When	NN	O	O
all	NN	O	O
the	NN	O	O
asthmatic	NN	O	O
patients	NN	O	O
were	NN	O	O
analyzed	NN	O	O
together	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
significant	NN	O	O
correlation	NN	O	O
between	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
the	NN	O	O
clinical	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
prednisolone	NN	O	O
(	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0.01	NN	O	O
)	NN	O	O
.	NN	O	O

No	NN	O	O
differences	NN	O	O
were	NN	O	O
observed	NN	O	O
between	NN	O	O
six	NN	O	O
of	NN	O	O
the	NN	O	O
sensitive	NN	O	O
and	NN	O	O
resistant	NN	O	O
patients	NN	O	O
in	NN	O	O
the	NN	O	O
clearance	NN	O	O
of	NN	O	O
plasma	NN	O	O
prednisolone	NN	O	O
derived	NN	O	O
from	NN	O	O
orally	NN	O	O
administered	NN	O	O
prednisone	NN	O	O
.	NN	O	O

Peripheral	NN	O	B-protein
blood	NN	O	I-protein
mononuclear	NN	O	I-protein
cell	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
were	NN	O	O
also	NN	O	O
characterized	NN	O	O
in	NN	O	O
five	NN	O	O
sensitive	NN	O	O
and	NN	O	O
seven	NN	O	O
resistant	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
numbers	NN	O	O
and	NN	O	O
binding	NN	O	O
affinities	NN	O	O
of	NN	O	O
these	NN	O	O
receptors	NN	O	B-protein
could	NN	O	O
not	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
observed	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
susceptibility	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
to	NN	O	O
functional	NN	O	O
inhibition	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
clinical	NN	O	O
glucocorticoid	NN	O	O
resistance	NN	O	O
in	NN	O	O
chronic	NN	O	O
asthma	NN	O	O
does	NN	O	O
not	NN	O	O
reflect	NN	O	O
abnormal	NN	O	O
glucocorticoid	NN	O	O
clearance	NN	O	O
but	NN	O	O
may	NN	O	O
be	NN	O	O
due	NN	O	O
at	NN	O	O
least	NN	O	O
partly	NN	O	O
to	NN	O	O
a	NN	O	O
relative	NN	O	O
insensitivity	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

This	NN	O	O
lack	NN	O	O
of	NN	O	O
sensitivity	NN	O	O
is	NN	O	O
unexplained	NN	O	O
but	NN	O	O
is	NN	O	O
not	NN	O	O
attributable	NN	O	O
to	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	B-DNA
(	NN	O	O
AP-1	NN	O	B-DNA
)	NN	O	O
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
treatment	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
myeloid	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	I-DNA
c-fos	NN	O	I-DNA
early	NN	O	I-DNA
response	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
present	NN	O	O
work	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
dexamethasone	NN	O	O
inhibits	NN	O	O
TPA-induced	NN	O	O
increases	NN	O	O
in	NN	O	O
c-jun	NN	O	O
and	NN	O	O
c-fos	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
in	NN	O	O
U-937	NN	O	B-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
findings	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
block	NN	O	O
in	NN	O	O
appearance	NN	O	O
of	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_type
phenotype	NN	O	I-cell_type
,	NN	O	O
including	NN	O	O
inhibition	NN	O	O
of	NN	O	O
TPA-induced	NN	O	O
increases	NN	O	O
in	NN	O	O
lamin	NN	O	B-RNA
A	NN	O	I-RNA
,	NN	O	I-RNA
lamin	NN	O	I-RNA
C	NN	O	I-RNA
,	NN	O	I-RNA
and	NN	O	I-RNA
vimentin	NN	O	I-RNA
transcripts	NN	O	I-RNA
.	NN	O	O

Other	NN	O	O
studies	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
TPA-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
c-fos	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
myeloid	NN	O	B-cell_type
leukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
finding	NN	O	O
that	NN	O	O
dexamethasone	NN	O	O
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
TPA-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
suggests	NN	O	O
that	NN	O	O
this	NN	O	O
glucocorticoid	NN	O	O
inhibits	NN	O	O
signals	NN	O	O
downstream	NN	O	O
or	NN	O	O
parallel	NN	O	O
to	NN	O	O
this	NN	O	O
enzyme	NN	O	B-protein
.	NN	O	O

Nuclear	NN	O	O
run-on	NN	O	O
assays	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
:	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
induction	NN	O	O
of	NN	O	O
c-jun	NN	O	O
and	NN	O	O
c-fos	NN	O	O
expression	NN	O	O
by	NN	O	O
TPA	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
transcriptional	NN	O	O
mechanisms	NN	O	O
,	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
TPA-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
c-fos	NN	O	B-protein
does	NN	O	O
not	NN	O	O
require	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
TPA-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
both	NN	O	O
genes	NN	O	O
is	NN	O	O
inhibited	NN	O	O
at	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
level	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
define	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
at	NN	O	O
the	NN	O	O
molecular	NN	O	O
level	NN	O	O
,	NN	O	O
we	NN	O	O
prepared	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
deleted	NN	O	O
c-jun	NN	O	B-DNA
promoter	NN	O	I-DNA
fragments	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
the	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Increases	NN	O	O
in	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
during	NN	O	O
transient	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
constructs	NN	O	O
in	NN	O	O
TPA-treated	NN	O	B-cell_line
U-937	NN	O	I-cell_line
cells	NN	O	I-cell_line
could	NN	O	O
be	NN	O	O
assigned	NN	O	O
to	NN	O	O
the	NN	O	O
region	NN	O	O
(	NN	O	O
-97	NN	O	O
to	NN	O	O
-20	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
that	NN	O	O
contains	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
induction	NN	O	O
of	NN	O	O
CAT	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
sensitive	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
dexamethasone	NN	O	O
down-regulates	NN	O	O
TPA-induced	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
c-jun	NN	O	B-DNA
gene	NN	O	O
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Tumor	NN	O	B-RNA
necrosis	NN	O	I-RNA
factor-alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
myelomonocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Role	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
by	NN	O	O
DNA	NN	O	B-DNA
sequence	NN	O	I-DNA
motifs	NN	O	I-DNA
and	NN	O	O
mRNA	NN	O	O
stabilization	NN	O	O
.	NN	O	O

The	NN	O	O
cytokine	NN	O	B-protein
TNF	NN	O	B-protein
mediates	NN	O	O
many	NN	O	O
of	NN	O	O
the	NN	O	O
pathologic	NN	O	O
signs	NN	O	O
of	NN	O	O
cachexia	NN	O	O
,	NN	O	O
inflammation	NN	O	O
,	NN	O	O
and	NN	O	O
sepsis	NN	O	O
.	NN	O	O

The	NN	O	O
current	NN	O	O
work	NN	O	O
describes	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
TNF	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_line
myelomonocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
after	NN	O	O
PMA	NN	O	O
stimulation	NN	O	O
.	NN	O	O

The	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
exhibit	NN	O	O
a	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
constitutive	NN	O	O
TNF	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
.	NN	O	O

Within	NN	O	O
2	NN	O	O
to	NN	O	O
4	NN	O	O
h	NN	O	O
of	NN	O	O
PMA	NN	O	O
exposure	NN	O	O
,	NN	O	O
steady	NN	O	O
state	NN	O	O
levels	NN	O	O
of	NN	O	O
TNF	NN	O	B-RNA
mRNA	NN	O	I-RNA
are	NN	O	O
markedly	NN	O	O
elevated	NN	O	O
in	NN	O	O
all	NN	O	O
myelomonocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
studied	NN	O	O
.	NN	O	O

This	NN	O	O
rise	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
increased	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
,	NN	O	O
which	NN	O	O
increased	NN	O	O
by	NN	O	O
almost	NN	O	O
twofold	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
an	NN	O	O
overall	NN	O	O
increase	NN	O	O
in	NN	O	O
transcription	NN	O	O
,	NN	O	O
which	NN	O	O
rises	NN	O	O
by	NN	O	O
more	NN	O	O
than	NN	O	O
sixfold	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
genomic	NN	O	B-DNA
TNF	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
DNase	NN	O	B-DNA
I	NN	O	I-DNA
hypersensitive	NN	O	I-DNA
site	NN	O	I-DNA
is	NN	O	O
detected	NN	O	O
within	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
promoter	NN	O	I-DNA
between	NN	O	O
-200	NN	O	O
to	NN	O	O
-100	NN	O	O
bp	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Although	NN	O	O
absent	NN	O	O
in	NN	O	O
nonexpressing	NN	O	B-cell_line
erythroleukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
DNase	NN	O	B-DNA
I	NN	O	I-DNA
site	NN	O	I-DNA
is	NN	O	O
present	NN	O	O
in	NN	O	O
uninduced	NN	O	B-cell_line
myelomonocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
is	NN	O	O
not	NN	O	O
changed	NN	O	O
after	NN	O	O
PMA	NN	O	O
induction	NN	O	O
.	NN	O	O

The	NN	O	O
PMA	NN	O	O
induction	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
correlated	NN	O	O
well	NN	O	O
with	NN	O	O
TNF	NN	O	B-protein
gene	NN	O	O
induction	NN	O	O
;	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
encoding	NN	O	O
other	NN	O	O
proteins	NN	O	O
in	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
complex	NN	O	I-protein
(	NN	O	O
junB	NN	O	B-protein
and	NN	O	O
junD	NN	O	B-protein
)	NN	O	O
were	NN	O	O
also	NN	O	O
induced	NN	O	O
by	NN	O	O
PMA	NN	O	O
.	NN	O	O

The	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
resting	NN	O	O
and	NN	O	O
induced	NN	O	B-cell_line
ML-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
contain	NN	O	O
proteins	NN	O	O
binding	NN	O	O
specifically	NN	O	O
to	NN	O	O
the	NN	O	O
AP-1	NN	O	B-DNA
,	NN	O	I-DNA
AP-2	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
NF	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
sequence	NN	O	I-DNA
located	NN	O	O
within	NN	O	O
the	NN	O	O
TNF	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

PMA	NN	O	O
induction	NN	O	O
increases	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
specific	NN	O	B-protein
binding	NN	O	I-protein
complexes	NN	O	I-protein
relative	NN	O	O
to	NN	O	O
the	NN	O	O
resting	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
regulatory	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
and	NN	O	I-DNA
murine	NN	O	I-DNA
TNF	NN	O	I-DNA
genes	NN	O	I-DNA
are	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Enhancement	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
1	NN	O	O
replication	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
by	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxycholecalciferol	NN	O	O
.	NN	O	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
(	NN	O	O
HIV	NN	O	O
)	NN	O	O
expression	NN	O	O
and	NN	O	O
replication	NN	O	O
are	NN	O	O
under	NN	O	O
tight	NN	O	O
regulatory	NN	O	O
control	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxycholecalciferol	NN	O	O
[	NN	O	O
1	NN	O	O
,	NN	O	O
25-	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
]	NN	O	O
enhances	NN	O	O
the	NN	O	O
replication	NN	O	O
of	NN	O	O
monocyte-	NN	O	O
and	NN	O	O
lymphocyte-tropic	NN	O	O
strains	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
up	NN	O	O
to	NN	O	O
10	NN	O	O
,	NN	O	O
000-fold	NN	O	O
in	NN	O	O
monocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
unfractionated	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
is	NN	O	O
therefore	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
potent	NN	O	O
regulators	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
described	NN	O	O
to	NN	O	O
date	NN	O	O
.	NN	O	O

Precursors	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
enhance	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
proportion	NN	O	O
to	NN	O	O
their	NN	O	O
affinity	NN	O	O
for	NN	O	O
the	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
intracellular	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
influences	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
by	NN	O	O
mechanisms	NN	O	O
involving	NN	O	O
this	NN	O	O
receptor	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
may	NN	O	O
have	NN	O	O
important	NN	O	O
implications	NN	O	O
for	NN	O	O
the	NN	O	O
design	NN	O	O
of	NN	O	O
effective	NN	O	O
therapy	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
suppresses	NN	O	O
human	NN	O	B-protein
B	NN	O	I-protein
lymphocyte	NN	O	I-protein
Ig	NN	O	I-protein
production	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
synthesis	NN	O	O
and	NN	O	O
the	NN	O	O
switch	NN	O	O
from	NN	O	O
the	NN	O	O
membrane	NN	O	B-RNA
form	NN	O	I-RNA
to	NN	O	O
the	NN	O	O
secreted	NN	O	B-RNA
form	NN	O	I-RNA
of	NN	O	O
Ig	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Transforming	NN	O	B-protein
growth	NN	O	I-protein
factor-beta	NN	O	I-protein
(	NN	O	O
TGF-beta	NN	O	B-protein
)	NN	O	O
inhibits	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
Ig	NN	O	B-protein
secretion	NN	O	O
and	NN	O	O
reduces	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
membrane	NN	O	O
Ig	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
TGF-beta	NN	O	B-protein
to	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
cultures	NN	O	I-cell_line
stimulated	NN	O	O
with	NN	O	O
Staphylococcus	NN	O	O
aureus	NN	O	O
Cowan	NN	O	O
strain	NN	O	O
I	NN	O	O
and	NN	O	O
IL-2	NN	O	B-protein
completely	NN	O	O
inhibited	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
Ig	NN	O	B-protein
secretion	NN	O	O
(	NN	O	O
greater	NN	O	O
than	NN	O	O
90	NN	O	O
%	NN	O	O
)	NN	O	O
and	NN	O	O
decreased	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
surface	NN	O	O
IgM	NN	O	O
,	NN	O	O
IgD	NN	O	O
,	NN	O	O
kappa	NN	O	O
L	NN	O	O
chain	NN	O	O
,	NN	O	O
and	NN	O	O
lambda	NN	O	O
L	NN	O	O
chain	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
TGF-beta	NN	O	B-protein
had	NN	O	O
only	NN	O	O
minimal	NN	O	O
effects	NN	O	O
on	NN	O	O
two	NN	O	O
other	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
membrane	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
HLA-DR	NN	O	B-protein
and	NN	O	O
CD20	NN	O	B-protein
.	NN	O	O

Internal	NN	O	O
labeling	NN	O	O
with	NN	O	O
[	NN	O	O
35S	NN	O	O
]	NN	O	O
methionine	NN	O	O
and	NN	O	O
immunoprecipitation	NN	O	O
with	NN	O	O
anti-IgM	NN	O	B-protein
,	NN	O	O
anti-kappa	NN	O	B-protein
,	NN	O	O
and	NN	O	O
anti-lambda	NN	O	B-protein
antibodies	NN	O	I-protein
revealed	NN	O	O
a	NN	O	O
striking	NN	O	O
reduction	NN	O	O
in	NN	O	O
kappa	NN	O	B-protein
L	NN	O	I-protein
chain	NN	O	I-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
TGF-beta	NN	O	B-protein
.	NN	O	O

A	NN	O	O
less	NN	O	O
pronounced	NN	O	O
reduction	NN	O	O
in	NN	O	O
lambda	NN	O	B-protein
L	NN	O	I-protein
chain	NN	O	I-protein
and	NN	O	O
microH	NN	O	B-protein
chain	NN	O	I-protein
was	NN	O	O
also	NN	O	O
noted	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
RNA	NN	O	O
purified	NN	O	O
from	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
TGF-beta	NN	O	B-protein
for	NN	O	O
varying	NN	O	O
time	NN	O	O
intervals	NN	O	O
revealed	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
in	NN	O	O
steady	NN	O	O
state	NN	O	O
kappa	NN	O	O
and	NN	O	O
lambda	NN	O	O
L	NN	O	O
chain	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
switch	NN	O	O
from	NN	O	O
the	NN	O	O
membrane	NN	O	B-RNA
forms	NN	O	I-RNA
of	NN	O	O
mu	NN	O	B-RNA
and	NN	O	O
gamma	NN	O	B-RNA
to	NN	O	O
their	NN	O	O
respective	NN	O	O
secreted	NN	O	B-RNA
forms	NN	O	I-RNA
was	NN	O	O
noted	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
TGF-beta	NN	O	B-protein
.	NN	O	O

Nuclear	NN	O	O
run-on	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
decreased	NN	O	O
transcription	NN	O	O
of	NN	O	O
kappa	NN	O	B-protein
L	NN	O	I-protein
chain	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
TGF-beta	NN	O	B-protein
on	NN	O	O
two	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
Oct-2	NN	O	B-protein
and	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
Ig	NN	O	B-protein
gene	NN	O	O
transcription	NN	O	O
were	NN	O	O
examined	NN	O	O
.	NN	O	O

Oct-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
and	NN	O	O
both	NN	O	O
Oct-2	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
were	NN	O	O
not	NN	O	O
altered	NN	O	O
by	NN	O	O
treatment	NN	O	O
with	NN	O	O
TGF-beta	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
factor	NN	O	I-protein
AP-1	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
not	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cell	NN	O	I-cell_type
Ig	NN	O	B-protein
production	NN	O	O
,	NN	O	O
were	NN	O	O
reduced	NN	O	O
by	NN	O	O
TGF-beta	NN	O	B-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
TGF-beta	NN	O	B-protein
decreases	NN	O	O
B	NN	O	O
lymphocyte	NN	O	O
Ig	NN	O	B-protein
secretion	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
Ig	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
switch	NN	O	O
from	NN	O	O
the	NN	O	O
membrane	NN	O	B-RNA
form	NN	O	I-RNA
to	NN	O	O
the	NN	O	O
secreted	NN	O	B-RNA
forms	NN	O	I-RNA
of	NN	O	O
mu	NN	O	B-RNA
and	NN	O	I-RNA
gamma	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
TGF-beta	NN	O	B-protein
inhibits	NN	O	O
Ig	NN	O	O
chain	NN	O	O
synthesis	NN	O	O
is	NN	O	O
unclear	NN	O	O
although	NN	O	O
it	NN	O	O
does	NN	O	O
not	NN	O	O
involve	NN	O	O
inhibition	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
or	NN	O	O
Oct-2	NN	O	B-protein
to	NN	O	O
their	NN	O	O
respective	NN	O	O
target	NN	O	O
sequences	NN	O	O
.	NN	O	O

-DOCSTART-	O

Murine	NN	O	B-protein
and	NN	O	I-protein
human	NN	O	I-protein
T-lymphocyte	NN	O	I-protein
GATA-3	NN	O	I-protein
factors	NN	O	I-protein
mediate	NN	O	O
transcription	NN	O	O
through	NN	O	O
a	NN	O	O
cis-regulatory	NN	O	B-DNA
element	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
T-cell	NN	O	I-DNA
receptor	NN	O	I-DNA
delta	NN	O	I-DNA
gene	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
family	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
activators	NN	O	I-protein
has	NN	O	O
recently	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
chickens	NN	O	O
;	NN	O	O
these	NN	O	O
transcriptional	NN	O	B-protein
activators	NN	O	I-protein
recognize	NN	O	O
a	NN	O	O
common	NN	O	O
consensus	NN	O	B-DNA
motif	NN	O	I-DNA
(	NN	O	O
WGATAR	NN	O	B-DNA
)	NN	O	O
through	NN	O	O
a	NN	O	O
conserved	NN	O	O
C4	NN	O	B-protein
zinc	NN	O	I-protein
finger	NN	O	I-protein
DNA-binding	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
members	NN	O	O
of	NN	O	O
this	NN	O	O
multigene	NN	O	O
family	NN	O	O
,	NN	O	O
cGATA-3	NN	O	B-protein
,	NN	O	O
is	NN	O	O
most	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
T-lymphocyte	NN	O	B-cell_type
cell	NN	O	I-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
human	NN	O	B-protein
and	NN	O	I-protein
murine	NN	O	I-protein
GATA-3	NN	O	I-protein
factors	NN	O	I-protein
shows	NN	O	O
a	NN	O	O
striking	NN	O	O
degree	NN	O	O
of	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
identity	NN	O	O
and	NN	O	O
similar	NN	O	O
patterns	NN	O	O
of	NN	O	O
tissue	NN	O	O
specificity	NN	O	O
of	NN	O	O
expression	NN	O	O
in	NN	O	O
these	NN	O	O
three	NN	O	O
organisms	NN	O	O
.	NN	O	O

The	NN	O	O
murine	NN	O	B-protein
and	NN	O	I-protein
human	NN	O	I-protein
factors	NN	O	I-protein
are	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
and	NN	O	I-cell_line
murine	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
can	NN	O	O
activate	NN	O	O
transcription	NN	O	O
through	NN	O	O
a	NN	O	O
tissue-specific	NN	O	B-DNA
GATA-binding	NN	O	I-DNA
site	NN	O	I-DNA
identified	NN	O	O
within	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
T-cell	NN	O	I-DNA
receptor	NN	O	I-DNA
delta	NN	O	I-DNA
gene	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
infer	NN	O	O
that	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
and	NN	O	I-protein
human	NN	O	I-protein
GATA-3	NN	O	I-protein
proteins	NN	O	I-protein
play	NN	O	O
a	NN	O	O
central	NN	O	O
and	NN	O	O
highly	NN	O	O
conserved	NN	O	O
role	NN	O	O
in	NN	O	O
vertebrate	NN	O	O
T-cell-specific	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Processing	NN	O	O
of	NN	O	O
the	NN	O	O
precursor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-protein
protease	NN	O	I-protein
during	NN	O	O
acute	NN	O	O
infection	NN	O	O
.	NN	O	O

Transcription	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type-1	NN	O	I-DNA
(	NN	O	I-DNA
HIV-1	NN	O	I-DNA
)	NN	O	I-DNA
genome	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
is	NN	O	O
stimulated	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
latently	NN	O	B-cell_type
infected	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

T-cell	NN	O	O
activation	NN	O	O
also	NN	O	O
correlates	NN	O	O
with	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
factor	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
which	NN	O	O
binds	NN	O	O
to	NN	O	O
two	NN	O	O
adjacent	NN	O	O
sites	NN	O	O
in	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
factor	NN	O	O
consists	NN	O	O
of	NN	O	O
two	NN	O	O
DNA-binding	NN	O	B-protein
subunits	NN	O	I-protein
of	NN	O	O
relative	NN	O	O
molecular	NN	O	O
mass	NN	O	O
50	NN	O	O
,	NN	O	O
000	NN	O	O
(	NN	O	O
50K	NN	O	O
)	NN	O	O
associated	NN	O	O
with	NN	O	O
two	NN	O	O
65K	NN	O	B-protein
subunits	NN	O	I-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
as	NN	O	O
an	NN	O	O
inactive	NN	O	B-protein
cytoplasmic	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

External	NN	O	O
stimuli	NN	O	O
,	NN	O	O
including	NN	O	O
those	NN	O	O
that	NN	O	O
activate	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
result	NN	O	O
in	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
active	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
cloning	NN	O	O
of	NN	O	O
the	NN	O	O
complementary	NN	O	B-DNA
DNA	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
50K	NN	O	B-protein
subunit	NN	O	I-protein
helped	NN	O	O
to	NN	O	O
identify	NN	O	O
an	NN	O	O
exclusively	NN	O	O
cytoplasmic	NN	O	B-protein
105K	NN	O	I-protein
precursor	NN	O	I-protein
(	NN	O	O
p105	NN	O	B-protein
)	NN	O	O
(	NN	O	O
V.B.	NN	O	O
,	NN	O	O
P.K.	NN	O	O
and	NN	O	O
A.I.	NN	O	O
,	NN	O	O
manuscript	NN	O	O
submitted	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
active	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
might	NN	O	O
therefore	NN	O	O
also	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
processing	NN	O	O
of	NN	O	O
p105	NN	O	B-protein
.	NN	O	O

Because	NN	O	O
HIV-1	NN	O	O
requires	NN	O	O
active	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
for	NN	O	O
efficient	NN	O	O
transcription	NN	O	O
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
on	NN	O	O
the	NN	O	O
processing	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
105K	NN	O	I-protein
precursor	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
protease	NN	O	O
can	NN	O	O
process	NN	O	O
p105	NN	O	B-protein
and	NN	O	O
increases	NN	O	O
levels	NN	O	O
of	NN	O	O
active	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

1	NN	O	B-RNA
,	NN	O	I-RNA
25-Dihydroxyvitamin	NN	O	I-RNA
D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
:	NN	O	O
expression	NN	O	O
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

1	NN	O	O
,	NN	O	O
25-Dihydroxyvitamin	NN	O	O
D3	NN	O	O
[	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
]	NN	O	O
induces	NN	O	O
differentiation	NN	O	O
and	NN	O	O
inhibits	NN	O	O
proliferation	NN	O	O
of	NN	O	O
myeloid	NN	O	B-cell_type
leukemic	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
various	NN	O	O
lines	NN	O	O
and	NN	O	O
patients	NN	O	O
;	NN	O	O
these	NN	O	O
effects	NN	O	O
are	NN	O	O
probably	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Little	NN	O	O
is	NN	O	O
known	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
in	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
examined	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
modulation	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
in	NN	O	O
various	NN	O	O
proliferating	NN	O	B-cell_type
and	NN	O	I-cell_type
nonproliferating	NN	O	I-cell_type
hematopoietic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
was	NN	O	O
detected	NN	O	O
in	NN	O	O
various	NN	O	O
kinds	NN	O	O
of	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
including	NN	O	O
macrophages	NN	O	O
and	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
cell	NN	O	O
lines	NN	O	O
KG-1	NN	O	B-cell_line
(	NN	O	O
myeloblasts	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
HL-60	NN	O	B-cell_line
(	NN	O	O
promyelocytes	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
ML-3	NN	O	B-cell_line
(	NN	O	O
myelomonoblasts	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
U937	NN	O	B-cell_line
,	NN	O	O
THP-1	NN	O	B-cell_line
(	NN	O	O
monoblasts	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
K562	NN	O	B-cell_line
(	NN	O	O
erythroblasts	NN	O	B-cell_line
)	NN	O	O
,	NN	O	O
and	NN	O	O
S-LB1	NN	O	B-cell_line
(	NN	O	O
HTLV-1-transfected	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
)	NN	O	O
.	NN	O	O

Receptor	NN	O	B-RNA
transcripts	NN	O	I-RNA
were	NN	O	O
4.6	NN	O	O
kilobases	NN	O	O
(	NN	O	O
kb	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
no	NN	O	O
variant	NN	O	O
sizes	NN	O	O
were	NN	O	O
observed	NN	O	O
.	NN	O	O

All	NN	O	O
cell	NN	O	O
lines	NN	O	O
examined	NN	O	O
in	NN	O	O
this	NN	O	O
group	NN	O	O
also	NN	O	O
expressed	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

Most	NN	O	O
B	NN	O	B-cell_line
lymphocyte	NN	O	I-cell_line
lines	NN	O	I-cell_line
expressed	NN	O	O
negligible	NN	O	O
levels	NN	O	O
of	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
;	NN	O	O
however	NN	O	O
;	NN	O	O
analysis	NN	O	O
of	NN	O	O
a	NN	O	O
lymphoid/myeloid	NN	O	B-cell_line
somatic	NN	O	I-cell_line
hybrid	NN	O	I-cell_line
suggested	NN	O	O
that	NN	O	O
suppression	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
may	NN	O	O
be	NN	O	O
a	NN	O	O
dominant	NN	O	O
characteristic	NN	O	O
.	NN	O	O

HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
cultured	NN	O	O
with	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
mol/L	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
for	NN	O	O
24	NN	O	O
to	NN	O	O
72	NN	O	O
hours	NN	O	O
,	NN	O	O
and	NN	O	O
levels	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptor	NN	O	I-protein
and	NN	O	O
its	NN	O	O
RNA	NN	O	O
were	NN	O	O
examined	NN	O	O
.	NN	O	O

Levels	NN	O	O
of	NN	O	O
RNA	NN	O	O
coding	NN	O	O
for	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
were	NN	O	O
not	NN	O	O
modulated	NN	O	O
by	NN	O	O
exposure	NN	O	O
to	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
ligand	NN	O	O
.	NN	O	O

Levels	NN	O	O
of	NN	O	O
occupied	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptor	NN	O	I-protein
protein	NN	O	I-protein
increased	NN	O	O
in	NN	O	O
these	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
;	NN	O	O
but	NN	O	O
the	NN	O	O
total	NN	O	O
number	NN	O	O
of	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptors	NN	O	I-protein
decreased	NN	O	O
about	NN	O	O
50	NN	O	O
%	NN	O	O
at	NN	O	O
24	NN	O	O
hours	NN	O	O
and	NN	O	O
returned	NN	O	O
toward	NN	O	O
normal	NN	O	O
at	NN	O	O
72	NN	O	O
hours	NN	O	O
.	NN	O	O

Steady-state	NN	O	O
levels	NN	O	O
of	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
were	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
terminal	NN	O	O
differentiation	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
toward	NN	O	O
either	NN	O	O
granulocytes	NN	O	B-cell_type
or	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

Nondividing	NN	O	O
macrophages	NN	O	B-cell_type
from	NN	O	O
normal	NN	O	O
individuals	NN	O	O
also	NN	O	O
expressed	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
nondividing	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
normal	NN	O	O
individuals	NN	O	O
did	NN	O	O
not	NN	O	O
express	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
;	NN	O	O
with	NN	O	O
stimulation	NN	O	O
of	NN	O	O
proliferation	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
accumulation	NN	O	O
of	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
increased	NN	O	O
markedly	NN	O	O
.	NN	O	O

Half-life	NN	O	O
(	NN	O	O
t1/2	NN	O	O
)	NN	O	O
of	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
was	NN	O	O
short	NN	O	O
(	NN	O	O
1	NN	O	O
hour	NN	O	O
)	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
measuring	NN	O	O
decay	NN	O	O
of	NN	O	O
the	NN	O	O
message	NN	O	O
after	NN	O	O
addition	NN	O	O
of	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
this	NN	O	O
short	NN	O	O
t1/2	NN	O	O
,	NN	O	O
accumulation	NN	O	O
of	NN	O	O
1	NN	O	B-RNA
,	NN	O	I-RNA
25	NN	O	I-RNA
(	NN	O	I-RNA
OH	NN	O	I-RNA
)	NN	O	I-RNA
2D3	NN	O	I-RNA
receptor	NN	O	I-RNA
RNA	NN	O	I-RNA
increased	NN	O	O
in	NN	O	O
cells	NN	O	O
as	NN	O	O
their	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
was	NN	O	O
inhibited	NN	O	O
.	NN	O	O

Further	NN	O	O
studies	NN	O	O
are	NN	O	O
required	NN	O	O
to	NN	O	O
understand	NN	O	O
the	NN	O	O
physiologic	NN	O	O
role	NN	O	O
of	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
proliferating	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	I-DNA
jun	NN	O	I-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
jun	NN	O	I-DNA
D	NN	O	I-DNA
proto-oncogenes	NN	O	I-DNA
in	NN	O	O
human	NN	O	B-cell_type
peripheral-blood	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
purified	NN	O	B-cell_type
human	NN	O	I-cell_type
peripheral-blood	NN	O	I-cell_type
granulocytes	NN	O	I-cell_type
express	NN	O	O
constitutively	NN	O	O
significant	NN	O	O
levels	NN	O	O
of	NN	O	O
proto-oncogenes	NN	O	B-DNA
c-jun	NN	O	B-RNA
,	NN	O	I-RNA
jun	NN	O	I-RNA
B	NN	O	I-RNA
and	NN	O	I-RNA
jun	NN	O	I-RNA
D	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Upon	NN	O	O
functional	NN	O	O
activation	NN	O	O
of	NN	O	O
granulocytes	NN	O	B-cell_type
by	NN	O	O
4	NN	O	O
beta-phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
,	NN	O	I-RNA
jun	NN	O	I-RNA
B	NN	O	I-RNA
and	NN	O	I-RNA
jun	NN	O	I-RNA
D	NN	O	I-RNA
transcripts	NN	O	I-RNA
were	NN	O	O
increased	NN	O	O
.	NN	O	O

The	NN	O	O
three	NN	O	O
jun	NN	O	B-DNA
genes	NN	O	I-DNA
showed	NN	O	O
a	NN	O	O
similar	NN	O	O
time	NN	O	O
course	NN	O	O
in	NN	O	O
their	NN	O	O
induction	NN	O	O
by	NN	O	O
PMA	NN	O	O
,	NN	O	O
maximal	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
being	NN	O	O
reached	NN	O	O
after	NN	O	O
60	NN	O	O
min	NN	O	O
of	NN	O	O
induction	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	O
jun	NN	O	B-DNA
B	NN	O	I-DNA
and	NN	O	O
jun	NN	O	B-DNA
D	NN	O	I-DNA
genes	NN	O	I-DNA
might	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
terminal	NN	O	B-cell_type
granulocyte	NN	O	I-cell_type
differentiation	NN	O	O
or	NN	O	O
in	NN	O	O
regulating	NN	O	O
granulocyte	NN	O	B-cell_type
functionality	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
29-kDa	NN	O	B-protein
proteins	NN	O	I-protein
phosphorylated	NN	O	O
in	NN	O	O
thrombin-activated	NN	O	B-cell_line
human	NN	O	I-cell_line
platelets	NN	O	I-cell_line
are	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
estrogen	NN	O	B-protein
receptor-related	NN	O	I-protein
27-kDa	NN	O	I-protein
heat	NN	O	I-protein
shock	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Thrombin	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
platelet	NN	O	O
activation	NN	O	O
,	NN	O	O
hemostasis	NN	O	O
,	NN	O	O
and	NN	O	O
thrombosis	NN	O	O
.	NN	O	O

Cellular	NN	O	O
activation	NN	O	O
by	NN	O	O
thrombin	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
multiple	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
most	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
unidentified	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
characterized	NN	O	O
several	NN	O	O
29-kDa	NN	O	B-protein
proteins	NN	O	I-protein
that	NN	O	O
are	NN	O	O
rapidly	NN	O	O
phosphorylated	NN	O	O
following	NN	O	O
exposure	NN	O	O
of	NN	O	O
intact	NN	O	B-cell_type
human	NN	O	I-cell_type
platelets	NN	O	I-cell_type
to	NN	O	O
thrombin	NN	O	B-protein
.	NN	O	O

A	NN	O	O
murine	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
raised	NN	O	O
to	NN	O	O
an	NN	O	O
unidentified	NN	O	B-protein
estrogen	NN	O	I-protein
receptor-related	NN	O	I-protein
29-kDa	NN	O	I-protein
protein	NN	O	I-protein
selectively	NN	O	O
recognized	NN	O	O
these	NN	O	O
proteins	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
more	NN	O	O
basic	NN	O	O
,	NN	O	O
unphosphorylated	NN	O	B-protein
27-kDa	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Cellular	NN	O	O
activation	NN	O	O
by	NN	O	O
thrombin	NN	O	B-protein
led	NN	O	O
to	NN	O	O
a	NN	O	O
marked	NN	O	O
shift	NN	O	O
in	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
protein	NN	O	O
from	NN	O	O
the	NN	O	O
27-kDa	NN	O	B-protein
unphosphorylated	NN	O	I-protein
form	NN	O	I-protein
to	NN	O	O
the	NN	O	O
29-kDa	NN	O	B-protein
phosphoprotein	NN	O	I-protein
species	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
this	NN	O	O
antibody	NN	O	O
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
and	NN	O	O
sequenced	NN	O	O
a	NN	O	O
human	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
encoding	NN	O	O
a	NN	O	O
protein	NN	O	O
that	NN	O	O
was	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
mammalian	NN	O	B-protein
27-kDa	NN	O	I-protein
heat	NN	O	I-protein
shock	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
HSP27	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
uncertain	NN	O	O
function	NN	O	O
that	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
phosphorylated	NN	O	O
to	NN	O	O
several	NN	O	O
forms	NN	O	O
and	NN	O	O
to	NN	O	O
be	NN	O	O
transcriptionally	NN	O	O
induced	NN	O	O
by	NN	O	O
estrogen	NN	O	O
.	NN	O	O

The	NN	O	O
29-kDa	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
confirmed	NN	O	O
to	NN	O	O
be	NN	O	O
phosphorylated	NN	O	O
forms	NN	O	O
of	NN	O	O
HSP27	NN	O	B-protein
by	NN	O	O
immunoprecipitation	NN	O	O
studies	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
``	NN	O	O
estrogen	NN	O	O
receptor-related	NN	O	O
protein	NN	O	O
''	NN	O	O
is	NN	O	O
HSP27	NN	O	B-protein
,	NN	O	O
and	NN	O	O
the	NN	O	O
three	NN	O	O
major	NN	O	O
29-kDa	NN	O	B-protein
proteins	NN	O	I-protein
phosphorylated	NN	O	O
in	NN	O	O
thrombin-activated	NN	O	B-cell_type
platelets	NN	O	I-cell_type
are	NN	O	O
forms	NN	O	O
of	NN	O	O
HSP27	NN	O	B-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
HSP27	NN	O	B-protein
in	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
events	NN	O	O
of	NN	O	O
platelet	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Isolation	NN	O	O
of	NN	O	O
a	NN	O	O
candidate	NN	O	O
repressor/activator	NN	O	O
,	NN	O	O
NF-E1	NN	O	B-protein
(	NN	O	O
YY-1	NN	O	B-protein
,	NN	O	O
delta	NN	O	B-protein
)	NN	O	O
,	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
kappa	NN	O	I-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
mu	NN	O	I-DNA
E1	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
determined	NN	O	O
that	NN	O	O
the	NN	O	O
developmental	NN	O	O
control	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
kappa	NN	O	I-DNA
3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
(	NN	O	O
kappa	NN	O	B-DNA
E3	NN	O	I-DNA
'	NN	O	I-DNA
)	NN	O	O
activity	NN	O	O
is	NN	O	O
the	NN	O	O
result	NN	O	O
of	NN	O	O
the	NN	O	O
combined	NN	O	O
influence	NN	O	O
of	NN	O	O
positive-	NN	O	B-DNA
and	NN	O	I-DNA
negative-acting	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
central	NN	O	O
core	NN	O	O
in	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
E3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
active	NN	O	O
at	NN	O	O
the	NN	O	O
pre-B-cell	NN	O	O
stage	NN	O	O
but	NN	O	O
is	NN	O	O
repressed	NN	O	O
by	NN	O	O
flanking	NN	O	B-DNA
negative-acting	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
negative-acting	NN	O	B-DNA
sequences	NN	O	I-DNA
repress	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
position-	NN	O	O
and	NN	O	O
orientation-independent	NN	O	O
manner	NN	O	O
at	NN	O	O
the	NN	O	O
pre-B-cell	NN	O	O
stage	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
isolated	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
cDNA	NN	O	I-DNA
clone	NN	O	I-DNA
encoding	NN	O	O
a	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
NF-E1	NN	O	B-protein
)	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
negative-acting	NN	O	B-DNA
segment	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
E3	NN	O	I-DNA
'	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
protein	NN	O	O
also	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
mu	NN	O	I-DNA
E1	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

NF-E1	NN	O	B-protein
is	NN	O	O
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
same	NN	O	O
gene	NN	O	O
as	NN	O	O
the	NN	O	O
YY-1	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
adeno-associated	NN	O	B-DNA
virus	NN	O	I-DNA
P5	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

NF-E1	NN	O	B-protein
is	NN	O	O
also	NN	O	O
the	NN	O	O
human	NN	O	B-protein
homologue	NN	O	I-protein
of	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
delta	NN	O	I-DNA
protein	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
ribosomal	NN	O	B-DNA
protein	NN	O	I-DNA
gene	NN	O	I-DNA
promoters	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
predicted	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
contains	NN	O	O
features	NN	O	O
characteristic	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
activators	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
transcriptional	NN	O	B-protein
repressors	NN	O	I-protein
.	NN	O	O

Cotransfection	NN	O	O
studies	NN	O	O
with	NN	O	O
this	NN	O	O
cDNA	NN	O	B-DNA
indicate	NN	O	O
that	NN	O	O
it	NN	O	O
can	NN	O	O
repress	NN	O	O
basal	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

The	NN	O	O
apparent	NN	O	O
dual	NN	O	O
function	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
human	NN	O	B-DNA
putative	NN	O	I-DNA
lymphocyte	NN	O	I-DNA
G0/G1	NN	O	I-DNA
switch	NN	O	I-DNA
gene	NN	O	I-DNA
containing	NN	O	O
a	NN	O	O
CpG-rich	NN	O	B-DNA
island	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
small	NN	O	B-protein
basic	NN	O	I-protein
protein	NN	O	I-protein
with	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
be	NN	O	O
phosphorylated	NN	O	O
.	NN	O	O

Genes	NN	O	O
actively	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
G0/G1	NN	O	B-DNA
switch	NN	O	I-DNA
(	NN	O	O
G0S	NN	O	B-RNA
genes	NN	O	I-RNA
)	NN	O	O
may	NN	O	O
be	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
during	NN	O	O
the	NN	O	O
lectin	NN	O	B-protein
-induced	NN	O	O
switch	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
G0	NN	O	O
to	NN	O	O
the	NN	O	O
G1	NN	O	O
phases	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
cycle	NN	O	O
.	NN	O	O

This	NN	O	O
paper	NN	O	O
presents	NN	O	O
studies	NN	O	O
of	NN	O	O
G0S2	NN	O	B-DNA
,	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
putative	NN	O	O
G0S	NN	O	B-RNA
genes	NN	O	I-RNA
,	NN	O	O
for	NN	O	O
which	NN	O	O
cDNAs	NN	O	B-DNA
were	NN	O	O
cloned	NN	O	O
and	NN	O	O
selected	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
differential	NN	O	O
cDNA	NN	O	O
hybridization	NN	O	O
.	NN	O	O

G0S2	NN	O	B-RNA
mRNA	NN	O	I-RNA
increases	NN	O	O
transiently	NN	O	O
within	NN	O	O
1-2	NN	O	O
hr	NN	O	O
of	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
lectin	NN	O	B-protein
or	NN	O	O
cycloheximide	NN	O	O
to	NN	O	O
cultured	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
a	NN	O	O
nearly	NN	O	B-DNA
full-length	NN	O	I-DNA
cDNA	NN	O	I-DNA
sequence	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
corresponding	NN	O	O
genomic	NN	O	B-DNA
sequence	NN	O	I-DNA
reveals	NN	O	O
one	NN	O	O
small	NN	O	O
intron	NN	O	O
and	NN	O	O
an	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
second	NN	O	O
exon	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
derived	NN	O	O
103-amino-acid	NN	O	B-protein
basic	NN	O	I-protein
protein	NN	O	I-protein
has	NN	O	O
two	NN	O	O
potential	NN	O	O
alpha-helical	NN	O	B-protein
domains	NN	O	I-protein
separated	NN	O	O
by	NN	O	O
a	NN	O	O
hydrophobic	NN	O	B-protein
region	NN	O	I-protein
with	NN	O	O
the	NN	O	O
potential	NN	O	O
to	NN	O	O
generate	NN	O	O
turns	NN	O	O
and	NN	O	O
assume	NN	O	O
a	NN	O	O
beta-sheet	NN	O	O
conformation	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
involvement	NN	O	O
in	NN	O	O
the	NN	O	O
G0/G1	NN	O	B-DNA
switch	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
protein	NN	O	O
contains	NN	O	O
potential	NN	O	O
sites	NN	O	O
for	NN	O	O
phosphorylation	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
and	NN	O	O
casein	NN	O	B-protein
kinase	NN	O	I-protein
II	NN	O	I-protein
.	NN	O	O

The	NN	O	O
gene	NN	O	O
contains	NN	O	O
a	NN	O	O
CpG-rich	NN	O	B-protein
island	NN	O	I-protein
suggesting	NN	O	O
expression	NN	O	O
in	NN	O	O
the	NN	O	O
germ	NN	O	O
line	NN	O	O
.	NN	O	O

An	NN	O	O
upstream	NN	O	B-DNA
segment	NN	O	I-DNA
contains	NN	O	O
tandem	NN	O	B-DNA
dinucleotide	NN	O	I-DNA
repeats	NN	O	I-DNA
(	NN	O	B-DNA
CT	NN	O	I-DNA
)	NN	O	I-DNA
19/	NN	O	I-DNA
(	NN	O	I-DNA
CA	NN	O	I-DNA
)	NN	O	I-DNA
16	NN	O	I-DNA
.	NN	O	O

There	NN	O	O
is	NN	O	O
a	NN	O	O
suitably	NN	O	O
located	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
potential	NN	O	O
sites	NN	O	O
for	NN	O	O
CCAAT-box	NN	O	B-protein
binding	NN	O	I-protein
factors	NN	O	I-protein
are	NN	O	O
far	NN	O	O
upstream	NN	O	O
,	NN	O	O
embedded	NN	O	O
in	NN	O	O
a	NN	O	O
42-nucleotide	NN	O	B-DNA
repeat	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Potential	NN	O	O
sites	NN	O	O
for	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
AP1	NN	O	B-protein
,	NN	O	O
AP2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
AP3	NN	O	B-protein
are	NN	O	O
consistent	NN	O	O
with	NN	O	O
rapid	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
inducing	NN	O	O
agents	NN	O	O
.	NN	O	O

-DOCSTART-	O

USF-related	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
HIV-TF1	NN	O	B-protein
,	NN	O	O
stimulates	NN	O	O
transcription	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus-1	NN	O	O
.	NN	O	O

The	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
HIV-TF1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
region	NN	O	O
about	NN	O	O
60	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
enhancer	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus-1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
,	NN	O	O
was	NN	O	O
purified	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

HIV-TF1	NN	O	B-protein
had	NN	O	O
a	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
39	NN	O	O
,	NN	O	O
000	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
HIV-TF1	NN	O	B-protein
to	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
activated	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
HIV-TF1-binding	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
was	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
site	NN	O	O
recognized	NN	O	O
by	NN	O	O
upstream	NN	O	B-protein
stimulatory	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
USF	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
adenovirus	NN	O	B-DNA
major	NN	O	I-DNA
late	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

DNA-binding	NN	O	O
properties	NN	O	O
of	NN	O	O
HIV-TF1	NN	O	B-protein
suggested	NN	O	O
that	NN	O	O
HIV-TF1	NN	O	B-protein
might	NN	O	O
be	NN	O	O
identical	NN	O	O
or	NN	O	O
related	NN	O	O
to	NN	O	O
USF	NN	O	B-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
treatment	NN	O	O
of	NN	O	O
purified	NN	O	O
HIV-TF1	NN	O	B-protein
by	NN	O	O
phosphatase	NN	O	B-protein
greatly	NN	O	O
reduced	NN	O	O
its	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
HIV-TF1	NN	O	B-protein
was	NN	O	O
essential	NN	O	O
for	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

The	NN	O	O
disruption	NN	O	O
of	NN	O	O
HIV-TF1-binding	NN	O	B-DNA
site	NN	O	I-DNA
induced	NN	O	O
a	NN	O	O
60	NN	O	O
%	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
HIV-TF1	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

-DOCSTART-	O

Severe	NN	O	O
5-fluorouracil	NN	O	O
toxicity	NN	O	O
secondary	NN	O	O
to	NN	O	O
dihydropyrimidine	NN	O	B-protein
dehydrogenase	NN	O	I-protein
deficiency	NN	O	O
.	NN	O	O

A	NN	O	O
potentially	NN	O	O
more	NN	O	O
common	NN	O	O
pharmacogenetic	NN	O	O
syndrome	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
describes	NN	O	O
the	NN	O	O
inheritance	NN	O	O
of	NN	O	O
a	NN	O	O
defect	NN	O	O
in	NN	O	O
pyrimidine	NN	O	O
catabolism	NN	O	O
and	NN	O	O
its	NN	O	O
association	NN	O	O
with	NN	O	O
drug-induced	NN	O	O
toxicity	NN	O	O
in	NN	O	O
a	NN	O	O
patient	NN	O	O
receiving	NN	O	O
5-fluorouracil	NN	O	O
(	NN	O	O
FUra	NN	O	O
)	NN	O	O
as	NN	O	O
adjuvant	NN	O	O
chemotherapy	NN	O	O
for	NN	O	O
breast	NN	O	O
carcinoma	NN	O	O
.	NN	O	O

The	NN	O	O
study	NN	O	O
population	NN	O	O
included	NN	O	O
the	NN	O	O
affected	NN	O	O
patient	NN	O	O
(	NN	O	O
proband	NN	O	O
)	NN	O	O
,	NN	O	O
nine	NN	O	O
of	NN	O	O
her	NN	O	O
blood	NN	O	O
relatives	NN	O	O
,	NN	O	O
and	NN	O	O
seven	NN	O	O
healthy	NN	O	O
volunteers	NN	O	O
.	NN	O	O

The	NN	O	O
activity	NN	O	O
of	NN	O	O
dihydropyrimidine	NN	O	B-protein
dehydrogenase	NN	O	I-protein
(	NN	O	O
DPD	NN	O	B-protein
)	NN	O	O
,	NN	O	O
the	NN	O	O
initial	NN	O	O
enzyme	NN	O	B-protein
of	NN	O	O
pyrimidine	NN	O	O
(	NN	O	O
and	NN	O	O
FUra	NN	O	O
)	NN	O	O
catabolism	NN	O	O
,	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
was	NN	O	O
measured	NN	O	O
in	NN	O	O
each	NN	O	O
subject	NN	O	O
by	NN	O	O
a	NN	O	O
specific	NN	O	O
radiometric	NN	O	O
assay	NN	O	O
using	NN	O	O
FUra	NN	O	O
as	NN	O	O
the	NN	O	O
substrate	NN	O	O
.	NN	O	O

The	NN	O	O
proband	NN	O	O
had	NN	O	O
no	NN	O	O
detectable	NN	O	O
DPD	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

When	NN	O	O
enzyme	NN	O	O
levels	NN	O	O
in	NN	O	O
the	NN	O	O
proband	NN	O	O
and	NN	O	O
relatives	NN	O	O
were	NN	O	O
compared	NN	O	O
with	NN	O	O
that	NN	O	O
in	NN	O	O
controls	NN	O	O
,	NN	O	O
an	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
pattern	NN	O	O
of	NN	O	O
inheritance	NN	O	O
was	NN	O	O
demonstrated	NN	O	O
.	NN	O	O

This	NN	O	O
is	NN	O	O
the	NN	O	O
third	NN	O	O
patient	NN	O	O
with	NN	O	O
severe	NN	O	O
FUra	NN	O	O
toxicity	NN	O	O
secondary	NN	O	O
to	NN	O	O
an	NN	O	O
alteration	NN	O	O
in	NN	O	O
pyrimidine	NN	O	O
catabolism	NN	O	O
and	NN	O	O
the	NN	O	O
second	NN	O	O
from	NN	O	O
our	NN	O	O
clinic	NN	O	O
population	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
this	NN	O	O
genetic	NN	O	O
defect	NN	O	O
may	NN	O	O
be	NN	O	O
greater	NN	O	O
than	NN	O	O
previously	NN	O	O
thought	NN	O	O
.	NN	O	O

Monitoring	NN	O	O
DPD	NN	O	B-protein
activity	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
the	NN	O	O
management	NN	O	O
of	NN	O	O
patients	NN	O	O
experiencing	NN	O	O
severe	NN	O	O
toxicity	NN	O	O
secondary	NN	O	O
to	NN	O	O
FUra	NN	O	O
chemotherapy	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
human	NN	O	O
herpesvirus	NN	O	O
6	NN	O	O
(	NN	O	O
HHV-6	NN	O	O
)	NN	O	O
strains	NN	O	O
GS	NN	O	O
and	NN	O	O
Z-29	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
identification	NN	O	O
of	NN	O	O
transactivating	NN	O	B-DNA
HHV-6	NN	O	I-DNA
(	NN	O	I-DNA
GS	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
fragments	NN	O	I-DNA
.	NN	O	O

Human	NN	O	O
herpesvirus	NN	O	O
6	NN	O	O
(	NN	O	O
HHV-6	NN	O	O
)	NN	O	O
can	NN	O	O
activate	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
HIV	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
accelerate	NN	O	O
cytopathic	NN	O	O
effects	NN	O	O
in	NN	O	O
HIV-infected	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
study	NN	O	O
examines	NN	O	O
the	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
promoter	NN	O	I-DNA
required	NN	O	O
for	NN	O	O
HHV-6	NN	O	O
transactivation	NN	O	O
in	NN	O	O
a	NN	O	O
heterogeneous	NN	O	O
population	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
or	NN	O	O
without	NN	O	O
antigenic	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Two	NN	O	O
different	NN	O	O
strains	NN	O	O
of	NN	O	O
HHV-6	NN	O	O
,	NN	O	O
GS	NN	O	O
and	NN	O	O
Z29	NN	O	O
,	NN	O	O
transactivated	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
GS	NN	O	O
strain	NN	O	O
transactivated	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
in	NN	O	O
both	NN	O	O
stimulated	NN	O	O
and	NN	O	O
resting	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
the	NN	O	O
Z29	NN	O	O
strain	NN	O	O
increased	NN	O	O
HIV	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
only	NN	O	O
in	NN	O	O
stimulated	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Three	NN	O	O
DNA	NN	O	O
clones	NN	O	O
containing	NN	O	O
HHV-6	NN	O	B-DNA
(	NN	O	I-DNA
GS	NN	O	I-DNA
)	NN	O	I-DNA
genomic	NN	O	I-DNA
fragments	NN	O	I-DNA
transactivated	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
cotransfected	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

A	NN	O	O
21.4-kb	NN	O	B-DNA
DNA	NN	O	I-DNA
clone	NN	O	I-DNA
,	NN	O	O
pZVB70	NN	O	B-DNA
,	NN	O	O
showed	NN	O	O
the	NN	O	O
highest	NN	O	O
transactivating	NN	O	O
ability	NN	O	O
,	NN	O	O
while	NN	O	O
two	NN	O	O
other	NN	O	O
DNA	NN	O	B-DNA
fragments	NN	O	I-DNA
,	NN	O	O
pZVB10	NN	O	B-DNA
(	NN	O	O
6.2	NN	O	O
kb	NN	O	O
)	NN	O	O
and	NN	O	O
pZVH14	NN	O	B-DNA
(	NN	O	O
8.7	NN	O	O
kb	NN	O	O
)	NN	O	O
,	NN	O	O
showed	NN	O	O
lower	NN	O	O
activity	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
clones	NN	O	O
,	NN	O	O
pZVH14	NN	O	B-DNA
,	NN	O	O
activated	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
promoter	NN	O	I-DNA
construct	NN	O	I-DNA
containing	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
mutated	NN	O	B-DNA
NF	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
promoter	NN	O	I-DNA
was	NN	O	O
not	NN	O	O
transactivated	NN	O	O
during	NN	O	O
HHV-6	NN	O	O
(	NN	O	O
GS	NN	O	O
)	NN	O	O
infection	NN	O	O
or	NN	O	O
after	NN	O	O
cotransfection	NN	O	O
with	NN	O	O
pZVB70	NN	O	B-DNA
or	NN	O	O
pZVB10	NN	O	B-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
sites	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
promoter	NN	O	I-DNA
are	NN	O	O
essential	NN	O	O
for	NN	O	O
its	NN	O	O
transactivation	NN	O	O
during	NN	O	O
HHV-6	NN	O	O
(	NN	O	O
GS	NN	O	O
)	NN	O	O
infection	NN	O	O
.	NN	O	O

By	NN	O	O
increasing	NN	O	O
HIV	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
HHV-6	NN	O	O
may	NN	O	O
consequently	NN	O	O
increase	NN	O	O
HIV	NN	O	O
replication	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
cytopathic	NN	O	O
effect	NN	O	O
on	NN	O	O
coinfected	NN	O	B-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
by	NN	O	O
cysteine	NN	O	O
and	NN	O	O
cysteine	NN	O	O
derivatives	NN	O	O
.	NN	O	O

HIV-1	NN	O	B-DNA
proviral	NN	O	I-DNA
DNA	NN	O	I-DNA
contains	NN	O	O
two	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

HIV-1-infected	NN	O	O
individuals	NN	O	O
have	NN	O	O
,	NN	O	O
on	NN	O	O
average	NN	O	O
,	NN	O	O
abnormally	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
and	NN	O	O
abnormally	NN	O	O
low	NN	O	O
plasma	NN	O	O
cysteine	NN	O	O
levels	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
investigated	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
cysteine	NN	O	O
and	NN	O	O
related	NN	O	O
thiols	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

The	NN	O	O
experiments	NN	O	O
in	NN	O	O
this	NN	O	O
report	NN	O	O
show	NN	O	O
that	NN	O	O
cysteine	NN	O	O
or	NN	O	O
N-acetylcysteine	NN	O	O
(	NN	O	O
NAC	NN	O	O
)	NN	O	O
raise	NN	O	O
the	NN	O	O
intracellular	NN	O	O
glutathione	NN	O	O
(	NN	O	O
GSH	NN	O	O
)	NN	O	O
level	NN	O	O
and	NN	O	O
inhibit	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
in	NN	O	O
persistently	NN	O	O
infected	NN	O	O
Molt-4	NN	O	B-cell_line
and	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
,	NN	O	O
inhibition	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
appears	NN	O	O
not	NN	O	O
to	NN	O	O
be	NN	O	O
directly	NN	O	O
correlated	NN	O	O
with	NN	O	O
GSH	NN	O	O
levels	NN	O	O
.	NN	O	O

Cysteine	NN	O	O
and	NN	O	O
NAC	NN	O	O
also	NN	O	O
inhibit	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
and	NN	O	O
chloramphenicol	NN	O	B-protein
acetyl-transferase	NN	O	I-protein
(	NN	O	O
CAT	NN	O	B-protein
)	NN	O	O
gene	NN	O	O
expression	NN	O	O
under	NN	O	O
control	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
uninfected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
cysteine	NN	O	O
deficiency	NN	O	O
in	NN	O	O
HIV-1-infected	NN	O	O
individuals	NN	O	O
may	NN	O	O
cause	NN	O	O
an	NN	O	O
over-expression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B-dependent	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
enhance	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

NAC	NN	O	O
may	NN	O	O
be	NN	O	O
considered	NN	O	O
for	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
HIV-1-infected	NN	O	O
individuals	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
thymus-specific	NN	O	B-protein
member	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HMG	NN	O	B-protein
protein	NN	O	I-protein
family	NN	O	I-protein
regulates	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
receptor	NN	O	I-DNA
C	NN	O	I-DNA
alpha	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
human	NN	O	B-protein
T	NN	O	I-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
plays	NN	O	O
a	NN	O	O
key	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
C	NN	O	O
alpha	NN	O	O
enhancer	NN	O	O
and	NN	O	O
binds	NN	O	O
to	NN	O	O
pyrimidine-rich	NN	O	B-DNA
elements	NN	O	I-DNA
(	NN	O	O
5'-PyCTTTG-3	NN	O	O
'	NN	O	O
)	NN	O	O
present	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
other	NN	O	O
T	NN	O	B-DNA
cell-specific	NN	O	I-DNA
control	NN	O	I-DNA
regions	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
information	NN	O	O
derived	NN	O	O
from	NN	O	O
the	NN	O	O
DNA	NN	O	B-protein
affinity-purified	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
we	NN	O	O
have	NN	O	O
now	NN	O	O
isolated	NN	O	O
cDNA	NN	O	B-DNA
clones	NN	O	I-DNA
encoding	NN	O	O
TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

The	NN	O	O
TCF-1	NN	O	B-DNA
alpha	NN	O	I-DNA
cDNA	NN	O	I-DNA
contains	NN	O	O
a	NN	O	O
single	NN	O	O
68-amino-acid	NN	O	B-protein
domain	NN	O	I-protein
that	NN	O	O
is	NN	O	O
homologous	NN	O	O
to	NN	O	O
a	NN	O	O
region	NN	O	O
conserved	NN	O	O
among	NN	O	O
high-mobility	NN	O	B-protein
group	NN	O	I-protein
(	NN	O	O
HMG	NN	O	B-protein
)	NN	O	O
and	NN	O	O
nonhistone	NN	O	B-protein
chromosomal	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
full-length	NN	O	B-DNA
and	NN	O	O
mutant	NN	O	B-DNA
cDNA	NN	O	I-DNA
clones	NN	O	I-DNA
in	NN	O	O
bacteria	NN	O	O
reveal	NN	O	O
that	NN	O	O
the	NN	O	O
single	NN	O	B-protein
HMG	NN	O	I-protein
motif	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
predicted	NN	O	O
to	NN	O	O
contain	NN	O	O
two	NN	O	O
extended	NN	O	O
alpha-helical	NN	O	B-protein
segments	NN	O	I-protein
,	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
direct	NN	O	O
the	NN	O	O
sequence-specific	NN	O	O
binding	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
to	NN	O	O
DNA	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
experiments	NN	O	O
demonstrate	NN	O	O
further	NN	O	O
that	NN	O	O
TCF-1	NN	O	B-RNA
alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
highly	NN	O	O
tissue	NN	O	O
specific	NN	O	O
,	NN	O	O
found	NN	O	O
primarily	NN	O	O
in	NN	O	O
the	NN	O	O
thymus	NN	O	B-cell_line
or	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
immature	NN	O	B-cell_line
CEM	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
expresses	NN	O	O
relatively	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-RNA
alpha	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
which	NN	O	O
are	NN	O	O
increased	NN	O	O
upon	NN	O	O
activation	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
cloned	NN	O	O
TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
protein	NN	O	I-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
TCR	NN	O	I-DNA
alpha	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
nonlymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
is	NN	O	O
strongly	NN	O	O
dependent	NN	O	O
on	NN	O	O
an	NN	O	O
additional	NN	O	O
T	NN	O	B-cell_line
cell-specific	NN	O	I-cell_line
protein	NN	O	I-cell_line
that	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
core	NN	O	O
enhancer	NN	O	O
.	NN	O	O

TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
is	NN	O	O
currently	NN	O	O
unique	NN	O	O
among	NN	O	O
the	NN	O	O
newly	NN	O	O
emerging	NN	O	O
family	NN	O	O
of	NN	O	O
DNA-binding	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
that	NN	O	O
share	NN	O	O
the	NN	O	O
HMG	NN	O	B-protein
motif	NN	O	I-protein
in	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
a	NN	O	O
highly	NN	O	B-protein
tissue-specific	NN	O	I-protein
RNA	NN	O	I-protein
polymerase	NN	O	I-protein
II	NN	O	I-protein
transcription	NN	O	B-protein
factor	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
HIV-1	NN	O	O
isolate	NN	O	O
containing	NN	O	O
alterations	NN	O	O
affecting	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Three	NN	O	O
molecular	NN	O	O
clones	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
derived	NN	O	O
from	NN	O	O
a	NN	O	O
single	NN	O	O
isolate	NN	O	O
(	NN	O	O
AL1	NN	O	O
)	NN	O	O
,	NN	O	O
exhibited	NN	O	O
distinct	NN	O	O
replicative	NN	O	O
and	NN	O	O
cytopathic	NN	O	O
properties	NN	O	O
during	NN	O	O
propagation	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
phenotypic	NN	O	O
differences	NN	O	O
observed	NN	O	O
were	NN	O	O
attributable	NN	O	O
,	NN	O	O
in	NN	O	O
large	NN	O	O
part	NN	O	O
,	NN	O	O
to	NN	O	O
changes	NN	O	O
affecting	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

Nucleotide	NN	O	O
sequence	NN	O	O
and	NN	O	O
PCR	NN	O	O
analyses	NN	O	O
demonstrated	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
novel	NN	O	O
duplications	NN	O	O
or	NN	O	O
deletions	NN	O	O
involving	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
were	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
original	NN	O	O
AL1	NN	O	O
virus	NN	O	O
stock	NN	O	O
.	NN	O	O

Subcloning	NN	O	O
of	NN	O	O
the	NN	O	O
variant	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
segments	NN	O	I-DNA
into	NN	O	O
LTR-driven	NN	O	B-DNA
CAT	NN	O	I-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
confirmed	NN	O	O
a	NN	O	O
correlation	NN	O	O
between	NN	O	O
promoter	NN	O	O
activity	NN	O	O
and	NN	O	O
replicative/cytopathic	NN	O	O
capacity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Multiple	NN	O	O
Oct2	NN	O	B-protein
isoforms	NN	O	I-protein
are	NN	O	O
generated	NN	O	O
by	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
.	NN	O	O

The	NN	O	O
interaction	NN	O	O
of	NN	O	O
the	NN	O	O
Oct2	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
with	NN	O	O
the	NN	O	O
cognate	NN	O	B-DNA
octamer	NN	O	I-DNA
motif	NN	O	I-DNA
ATGCAAAT	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
determinant	NN	O	O
of	NN	O	O
the	NN	O	O
lymphoid-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Ectopic	NN	O	O
expression	NN	O	O
of	NN	O	O
cloned	NN	O	B-DNA
Oct2	NN	O	I-DNA
cDNA	NN	O	I-DNA
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
sufficient	NN	O	O
to	NN	O	O
reconstitute	NN	O	O
at	NN	O	O
least	NN	O	O
some	NN	O	O
aspects	NN	O	O
of	NN	O	O
this	NN	O	O
regulation	NN	O	O
in	NN	O	O
non-lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
describe	NN	O	O
the	NN	O	O
isolation	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
multiple	NN	O	B-DNA
cDNAs	NN	O	I-DNA
encoding	NN	O	O
mouse	NN	O	B-protein
Oct2	NN	O	I-protein
from	NN	O	O
a	NN	O	O
mature	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
isoforms	NN	O	O
of	NN	O	O
this	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
generated	NN	O	O
from	NN	O	O
a	NN	O	O
single	NN	O	O
gene	NN	O	O
by	NN	O	O
an	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
mechanism	NN	O	O
.	NN	O	O

All	NN	O	O
the	NN	O	O
isoforms	NN	O	B-protein
retain	NN	O	O
the	NN	O	O
previously	NN	O	O
characterized	NN	O	O
POU-domain	NN	O	O
and	NN	O	O
are	NN	O	O
therefore	NN	O	O
able	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
.	NN	O	O

Different	NN	O	O
amounts	NN	O	O
of	NN	O	O
the	NN	O	O
various	NN	O	O
isoforms	NN	O	B-protein
are	NN	O	O
present	NN	O	O
within	NN	O	O
the	NN	O	O
same	NN	O	O
B-cell	NN	O	B-cell_type
regardless	NN	O	O
of	NN	O	O
the	NN	O	O
developmental	NN	O	O
stage	NN	O	O
of	NN	O	O
B-cell	NN	O	B-cell_type
differentiation	NN	O	O
and	NN	O	O
at	NN	O	O
least	NN	O	O
some	NN	O	O
of	NN	O	O
the	NN	O	O
isoforms	NN	O	B-protein
are	NN	O	O
conserved	NN	O	O
between	NN	O	O
mouse	NN	O	O
and	NN	O	O
humans	NN	O	O
.	NN	O	O

In	NN	O	O
cotransfection	NN	O	O
experiments	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
all	NN	O	O
the	NN	O	O
isoforms	NN	O	B-protein
are	NN	O	O
able	NN	O	O
to	NN	O	O
activate	NN	O	O
an	NN	O	O
octamer	NN	O	B-DNA
containing	NN	O	O
promoter	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
fibroblasts	NN	O	O
revealing	NN	O	O
an	NN	O	O
unexpected	NN	O	O
functional	NN	O	O
redundancy	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
isoforms	NN	O	B-protein
encodes	NN	O	O
the	NN	O	O
previously	NN	O	O
described	NN	O	O
lymphoid-specific	NN	O	B-protein
Oct2B	NN	O	I-protein
protein	NN	O	I-protein
which	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
(	NN	O	I-DNA
IgH	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Regulation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
cholesterol	NN	O	O
synthesis	NN	O	O
in	NN	O	O
hypercholesterolemia	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
]	NN	O	O

The	NN	O	O
rate	NN	O	O
of	NN	O	O
endogenous	NN	O	O
cholesterol	NN	O	O
synthesis	NN	O	O
in	NN	O	O
blood	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
skin	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
type	NN	O	O
IIa	NN	O	O
hyperlipidemia	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
increased	NN	O	O
in	NN	O	O
comparison	NN	O	O
with	NN	O	O
healthy	NN	O	O
donors	NN	O	O
.	NN	O	O

The	NN	O	O
cells	NN	O	O
of	NN	O	O
hyperlipidemic	NN	O	O
patients	NN	O	O
had	NN	O	O
lowered	NN	O	O
levels	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
concomitantly	NN	O	O
with	NN	O	O
a	NN	O	O
partial	NN	O	O
loss	NN	O	O
of	NN	O	O
their	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

In	NN	O	O
fibroblasts	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
hereditary	NN	O	O
hypercholesteremia	NN	O	O
of	NN	O	O
homozygous	NN	O	O
type	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
did	NN	O	O
not	NN	O	O
exceed	NN	O	O
10	NN	O	O
%	NN	O	O
of	NN	O	O
their	NN	O	O
content	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
decrease	NN	O	O
of	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
patients	NN	O	O
with	NN	O	O
type	NN	O	O
IIa	NN	O	O
hyperlipidemia	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
compensatory	NN	O	O
response	NN	O	O
of	NN	O	O
cells	NN	O	O
culminating	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
endogenous	NN	O	O
cholesterol	NN	O	O
synthesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	O
:	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
65-kDa	NN	O	B-protein
plus	NN	O	O
50-kDa	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
dimer	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
monocytic	NN	O	O
cells	NN	O	O
can	NN	O	O
provide	NN	O	O
a	NN	O	O
reservoir	NN	O	O
for	NN	O	O
viral	NN	O	O
production	NN	O	O
in	NN	O	O
vivo	NN	O	O
,	NN	O	O
their	NN	O	O
regulation	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
transcription	NN	O	O
can	NN	O	O
be	NN	O	O
either	NN	O	O
latent	NN	O	O
,	NN	O	O
restricted	NN	O	O
,	NN	O	O
or	NN	O	O
productive	NN	O	O
.	NN	O	O

These	NN	O	O
differences	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
molecularly	NN	O	O
defined	NN	O	O
.	NN	O	O

In	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
restricted	NN	O	O
HIV	NN	O	O
expression	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
DNA-protein	NN	O	B-protein
binding	NN	O	I-protein
complex	NN	O	I-protein
formation	NN	O	O
with	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
promoter-enhancer	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
markedly	NN	O	O
less	NN	O	O
viral	NN	O	B-RNA
RNA	NN	O	I-RNA
production	NN	O	O
.	NN	O	O

This	NN	O	O
absence	NN	O	O
of	NN	O	O
binding	NN	O	O
was	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
;	NN	O	O
the	NN	O	O
65-kDa	NN	O	B-protein
plus	NN	O	I-protein
50-kDa	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
heterodimer	NN	O	I-protein
was	NN	O	O
preferentially	NN	O	O
lost	NN	O	O
.	NN	O	O

Adding	NN	O	O
purified	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
protein	NN	O	I-protein
to	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
cells	NN	O	O
with	NN	O	O
restricted	NN	O	O
expression	NN	O	O
overcomes	NN	O	O
this	NN	O	O
lack	NN	O	O
of	NN	O	O
binding	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
treatment	NN	O	O
of	NN	O	O
these	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
with	NN	O	O
sodium	NN	O	O
deoxycholate	NN	O	O
restored	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
form	NN	O	O
the	NN	O	O
heterodimer	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
treatment	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
these	NN	O	O
cells	NN	O	O
that	NN	O	O
had	NN	O	O
restricted	NN	O	O
expression	NN	O	O
with	NN	O	O
lipopolysaccharide	NN	O	O
increased	NN	O	O
viral	NN	O	O
production	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activity	NN	O	O
.	NN	O	O

Antiserum	NN	O	O
specific	NN	O	O
for	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
c-rel-specific	NN	O	O
antiserum	NN	O	O
,	NN	O	O
disrupted	NN	O	O
heterodimer	NN	O	B-protein
complex	NN	O	I-protein
formation	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
both	NN	O	O
NF-kappa	NN	O	B-protein
B-binding	NN	O	I-protein
complexes	NN	O	I-protein
are	NN	O	O
needed	NN	O	O
for	NN	O	O
optimal	NN	O	O
viral	NN	O	O
transcription	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
the	NN	O	O
65-kDa	NN	O	B-protein
plus	NN	O	O
50-kDa	NN	O	B-protein
heterodimer	NN	O	I-protein
to	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
can	NN	O	O
be	NN	O	O
negatively	NN	O	O
regulated	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
providing	NN	O	O
one	NN	O	O
mechanism	NN	O	O
restricting	NN	O	O
HIV-1	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
normal	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
:	NN	O	O
effects	NN	O	O
of	NN	O	O
age	NN	O	O
,	NN	O	O
gender	NN	O	O
,	NN	O	O
season	NN	O	O
,	NN	O	O
and	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
.	NN	O	O

We	NN	O	O
measured	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
(	NN	O	O
MNL	NN	O	B-cell_type
)	NN	O	O
isolated	NN	O	O
from	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
of	NN	O	O
145	NN	O	O
apparently	NN	O	O
healthy	NN	O	O
volunteers	NN	O	O
(	NN	O	O
86	NN	O	O
men	NN	O	O
and	NN	O	O
59	NN	O	O
women	NN	O	O
)	NN	O	O
.	NN	O	O

An	NN	O	O
age-related	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
was	NN	O	O
suggested	NN	O	O
between	NN	O	O
subjects	NN	O	O
younger	NN	O	O
than	NN	O	O
20	NN	O	O
years	NN	O	O
and	NN	O	O
elderly	NN	O	O
subjects	NN	O	O
;	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
apparent	NN	O	O
seasonal	NN	O	O
variation	NN	O	O
in	NN	O	O
GR	NN	O	B-protein
.	NN	O	O

Gender	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
was	NN	O	O
not	NN	O	O
significant	NN	O	O
,	NN	O	O
although	NN	O	O
women	NN	O	O
showed	NN	O	O
slightly	NN	O	O
fewer	NN	O	O
GR	NN	O	B-protein
.	NN	O	O

Eight	NN	O	O
patients	NN	O	O
with	NN	O	O
dermatomyositis/polymyositis	NN	O	O
were	NN	O	O
examined	NN	O	O
to	NN	O	O
determine	NN	O	O
whether	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
MNL	NN	O	B-cell_type
could	NN	O	O
be	NN	O	O
down-regulated	NN	O	O
by	NN	O	O
their	NN	O	O
cognate	NN	O	O
ligands	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
MNL	NN	O	B-cell_type
from	NN	O	O
these	NN	O	O
patients	NN	O	O
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
one	NN	O	O
month	NN	O	O
after	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
prednisolone	NN	O	O
therapy	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
normal	NN	O	O
subjects	NN	O	O
,	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
MNL	NN	O	B-cell_type
did	NN	O	O
not	NN	O	O
demonstrate	NN	O	O
circadian	NN	O	O
variation	NN	O	O
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
concentrations	NN	O	O
of	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
.	NN	O	O

-DOCSTART-	O

Synergism	NN	O	O
between	NN	O	O
two	NN	O	O
distinct	NN	O	O
elements	NN	O	O
of	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
enhancer	NN	O	I-DNA
during	NN	O	O
activation	NN	O	O
by	NN	O	O
the	NN	O	O
trans-activator	NN	O	B-protein
of	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
conducted	NN	O	O
functional	NN	O	O
studies	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
elements	NN	O	I-DNA
of	NN	O	O
human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
using	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
Jurkat	NN	O	B-cell_line
and	NN	O	O
MOLT	NN	O	B-cell_line
4	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
are	NN	O	O
negative	NN	O	O
for	NN	O	O
HTLV-I	NN	O	O
,	NN	O	O
and	NN	O	O
MT-2	NN	O	O
and	NN	O	O
TL-Mor	NN	O	O
,	NN	O	O
which	NN	O	O
carry	NN	O	O
the	NN	O	O
proviral	NN	O	O
genome	NN	O	O
of	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

Two	NN	O	O
distinct	NN	O	O
elements	NN	O	O
have	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

One	NN	O	O
is	NN	O	O
the	NN	O	O
21-base-pair	NN	O	B-DNA
(	NN	O	I-DNA
bp	NN	O	I-DNA
)	NN	O	I-DNA
core	NN	O	I-DNA
element	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
trans-activation	NN	O	O
by	NN	O	O
the	NN	O	O
HTLV-I	NN	O	O
trans-activator	NN	O	B-protein
p40tax	NN	O	B-protein
and	NN	O	O
that	NN	O	O
has	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
cyclic-AMP	NN	O	B-protein
responsive	NN	O	I-protein
element	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	I-protein
CREB	NN	O	I-protein
)	NN	O	I-protein
-like	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
is	NN	O	O
a	NN	O	O
region	NN	O	O
interposed	NN	O	O
between	NN	O	O
the	NN	O	O
21-bp	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
a	NN	O	O
subfragment	NN	O	O
(	NN	O	O
C26	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
region	NN	O	O
between	NN	O	O
the	NN	O	O
21-bp	NN	O	B-DNA
elements	NN	O	I-DNA
is	NN	O	O
involved	NN	O	O
in	NN	O	O
trans-activation	NN	O	O
by	NN	O	O
p40tax	NN	O	B-protein
,	NN	O	O
possibly	NN	O	O
through	NN	O	O
binding	NN	O	O
to	NN	O	O
an	NN	O	O
NF-kappa	NN	O	B-protein
B-like	NN	O	I-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
or	NN	O	O
factors	NN	O	O
.	NN	O	O

Formation	NN	O	O
of	NN	O	O
the	NN	O	O
protein-DNA	NN	O	B-protein
complex	NN	O	I-protein
with	NN	O	O
the	NN	O	O
C26	NN	O	B-protein
subfragment	NN	O	I-protein
was	NN	O	O
positively	NN	O	O
affected	NN	O	O
by	NN	O	O
p40tax	NN	O	B-protein
.	NN	O	O

The	NN	O	O
C26	NN	O	B-DNA
element	NN	O	I-DNA
conferred	NN	O	O
partial	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
p40tax	NN	O	B-protein
when	NN	O	O
linked	NN	O	O
to	NN	O	O
one	NN	O	O
copy	NN	O	O
of	NN	O	O
the	NN	O	O
21-bp	NN	O	B-DNA
element	NN	O	I-DNA
that	NN	O	O
,	NN	O	O
by	NN	O	O
itself	NN	O	O
,	NN	O	O
showed	NN	O	O
little	NN	O	O
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
p40tax	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
C26	NN	O	B-DNA
element	NN	O	I-DNA
alone	NN	O	O
,	NN	O	O
even	NN	O	O
when	NN	O	O
repeated	NN	O	O
,	NN	O	O
could	NN	O	O
not	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
p40tax	NN	O	B-protein
,	NN	O	O
unlike	NN	O	O
other	NN	O	O
NF-kappa	NN	O	B-DNA
B-binding	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
C26	NN	O	B-DNA
element	NN	O	I-DNA
itself	NN	O	O
was	NN	O	O
profoundly	NN	O	O
activated	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
with	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
therefore	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
enhancer	NN	O	I-DNA
contains	NN	O	O
multiple	NN	O	B-DNA
functional	NN	O	I-DNA
elements	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
at	NN	O	O
least	NN	O	O
CREB-	NN	O	B-protein
and	NN	O	I-protein
NF-kappa	NN	O	I-protein
B-like	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
which	NN	O	O
synergistically	NN	O	O
cooperate	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
p40tax	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
TPA-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-DNA
enhancer	NN	O	I-DNA
may	NN	O	O
be	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
C26	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
gene	NN	O	O
regulation	NN	O	O
in	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
stimulated	NN	O	B-cell_line
T	NN	O	I-cell_line
and	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
minimal	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
tumor	NN	O	I-DNA
necrosis	NN	O	I-DNA
factor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
TNF-alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
necessary	NN	O	O
for	NN	O	O
its	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
and	NN	O	I-cell_line
B	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
has	NN	O	O
been	NN	O	O
localized	NN	O	O
between	NN	O	O
-52	NN	O	B-DNA
and	NN	O	I-DNA
+89	NN	O	I-DNA
nucleotides	NN	O	I-DNA
(	NN	O	O
nt	NN	O	O
)	NN	O	O
relative	NN	O	O
to	NN	O	O
the	NN	O	O
gene	NN	O	O
's	NN	O	O
transcriptional	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
these	NN	O	O
sequences	NN	O	O
to	NN	O	O
those	NN	O	O
required	NN	O	O
to	NN	O	O
mediate	NN	O	O
virus	NN	O	O
or	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
gene	NN	O	O
reveal	NN	O	O
significant	NN	O	O
differences	NN	O	O
,	NN	O	O
and	NN	O	O
thus	NN	O	O
,	NN	O	O
the	NN	O	O
sequence	NN	O	O
requirements	NN	O	O
for	NN	O	O
PMA	NN	O	O
induction	NN	O	O
are	NN	O	O
distinct	NN	O	O
from	NN	O	O
those	NN	O	O
that	NN	O	O
mediate	NN	O	O
induction	NN	O	O
by	NN	O	O
virus	NN	O	O
or	NN	O	O
LPS	NN	O	O
.	NN	O	O

Although	NN	O	O
three	NN	O	O
sites	NN	O	O
in	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
(	NN	O	O
kappa	NN	O	O
1	NN	O	O
,	NN	O	O
kappa	NN	O	O
2	NN	O	O
,	NN	O	O
and	NN	O	O
kappa	NN	O	O
3	NN	O	O
)	NN	O	O
specifically	NN	O	O
bind	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
lymphoid	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
by	NN	O	O
PMA	NN	O	O
does	NN	O	O
not	NN	O	O
correlate	NN	O	O
with	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
activities	NN	O	O
displayed	NN	O	O
by	NN	O	O
different	NN	O	O
T	NN	O	B-cell_line
and	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
kappa	NN	O	B-DNA
1-kappa	NN	O	I-DNA
3	NN	O	I-DNA
can	NN	O	O
each	NN	O	O
be	NN	O	O
deleted	NN	O	O
from	NN	O	O
the	NN	O	O
TNF-alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
with	NN	O	O
little	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
gene	NN	O	O
's	NN	O	O
inducibility	NN	O	O
by	NN	O	O
PMA	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
TNF-alpha	NN	O	B-RNA
mRNA	NN	O	I-RNA
induction	NN	O	O
by	NN	O	O
PMA	NN	O	O
,	NN	O	O
like	NN	O	O
its	NN	O	O
induction	NN	O	O
by	NN	O	O
virus	NN	O	O
and	NN	O	O
LPS	NN	O	O
,	NN	O	O
is	NN	O	O
not	NN	O	O
primarily	NN	O	O
mediated	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
but	NN	O	O
rather	NN	O	O
is	NN	O	O
mediated	NN	O	O
through	NN	O	O
other	NN	O	O
sequences	NN	O	O
and	NN	O	O
protein	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
multimers	NN	O	O
of	NN	O	O
kappa	NN	O	O
1-kappa	NN	O	O
3	NN	O	O
can	NN	O	O
confer	NN	O	O
PMA	NN	O	O
inducibility	NN	O	O
on	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
B	NN	O	B-cell_line
(	NN	O	I-cell_line
Raji	NN	O	I-cell_line
)	NN	O	I-cell_line
,	NN	O	I-cell_line
but	NN	O	I-cell_line
not	NN	O	I-cell_line
a	NN	O	I-cell_line
T	NN	O	I-cell_line
(	NN	O	I-cell_line
HUT78	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

However	NN	O	O
they	NN	O	O
are	NN	O	O
not	NN	O	O
functional	NN	O	O
on	NN	O	O
a	NN	O	O
truncated	NN	O	O
TNF-alpha	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
promoter	NN	O	O
context	NN	O	O
and	NN	O	O
cell	NN	O	O
type	NN	O	O
specificity	NN	O	O
influence	NN	O	O
the	NN	O	O
PMA	NN	O	O
inducible	NN	O	O
function	NN	O	O
of	NN	O	O
these	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Differentiation-associated	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
BZLF1	NN	O	I-protein
transactivator	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
oral	NN	O	O
hairy	NN	O	O
leukoplakia	NN	O	O
.	NN	O	O

The	NN	O	O
BZLF1	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
key	NN	O	O
immediate-early	NN	O	B-protein
protein	NN	O	I-protein
which	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
disrupt	NN	O	O
virus	NN	O	O
latency	NN	O	O
in	NN	O	O
EBV-infected	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
generated	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
,	NN	O	O
BZ1	NN	O	B-protein
,	NN	O	O
to	NN	O	O
BZLF1	NN	O	B-protein
which	NN	O	O
reacts	NN	O	O
in	NN	O	O
immunohistology	NN	O	O
,	NN	O	O
immunoblotting	NN	O	O
,	NN	O	O
and	NN	O	O
immunoprecipitation	NN	O	O
and	NN	O	O
which	NN	O	O
recognizes	NN	O	O
both	NN	O	O
the	NN	O	O
active	NN	O	B-protein
,	NN	O	I-protein
dimeric	NN	O	I-protein
form	NN	O	I-protein
and	NN	O	O
the	NN	O	O
inactive	NN	O	B-protein
,	NN	O	I-protein
monomeric	NN	O	I-protein
form	NN	O	I-protein
of	NN	O	O
the	NN	O	O
protein	NN	O	O
.	NN	O	O

Biopsies	NN	O	O
of	NN	O	O
oral	NN	O	O
hairy	NN	O	O
leukoplakia	NN	O	O
,	NN	O	O
an	NN	O	O
AIDS-associated	NN	O	O
lesion	NN	O	O
characterized	NN	O	O
by	NN	O	O
high-level	NN	O	O
EBV	NN	O	O
replication	NN	O	O
,	NN	O	O
were	NN	O	O
examined	NN	O	O
by	NN	O	O
immunohistochemistry	NN	O	O
using	NN	O	O
the	NN	O	O
BZ1	NN	O	B-protein
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

A	NN	O	O
differentiation-associated	NN	O	O
pattern	NN	O	O
of	NN	O	O
BZLF1	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
observed	NN	O	O
,	NN	O	O
BZ1	NN	O	B-protein
reacting	NN	O	O
with	NN	O	O
nuclei	NN	O	O
of	NN	O	O
the	NN	O	O
upper	NN	O	O
spinous	NN	O	O
layer	NN	O	O
of	NN	O	O
the	NN	O	O
lesion	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
BZLF1	NN	O	B-DNA
promoter	NN	O	I-DNA
may	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
squamous	NN	O	O
differentiation	NN	O	O
.	NN	O	O

A	NN	O	O
comparison	NN	O	O
of	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
to	NN	O	O
EBV	NN	O	B-DNA
DNA	NN	O	I-DNA
and	NN	O	O
viral	NN	O	B-protein
capsid	NN	O	I-protein
antigen	NN	O	I-protein
staining	NN	O	O
with	NN	O	O
BZ1	NN	O	B-protein
reactivity	NN	O	O
suggested	NN	O	O
that	NN	O	O
BZLF1	NN	O	B-protein
expression	NN	O	O
precedes	NN	O	O
rampant	NN	O	O
virus	NN	O	O
replication	NN	O	O
.	NN	O	O

The	NN	O	O
inability	NN	O	O
to	NN	O	O
detect	NN	O	O
EBV	NN	O	O
in	NN	O	O
the	NN	O	O
lower	NN	O	O
epithelial	NN	O	O
layers	NN	O	O
of	NN	O	O
oral	NN	O	O
hairy	NN	O	O
leukoplakia	NN	O	O
raises	NN	O	O
questions	NN	O	O
concerning	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
EBV	NN	O	O
latency	NN	O	O
and	NN	O	O
persistence	NN	O	O
in	NN	O	O
stratified	NN	O	O
squamous	NN	O	O
epithelium	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Changes	NN	O	O
in	NN	O	O
levels	NN	O	O
of	NN	O	O
leucocytic	NN	O	B-protein
estrogen	NN	O	I-protein
receptor	NN	O	I-protein
in	NN	O	O
patients	NN	O	O
with	NN	O	O
menopausal	NN	O	O
type	NN	O	O
II	NN	O	O
diabetes	NN	O	O
and	NN	O	O
its	NN	O	O
significance	NN	O	O
]	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
estrogen	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
in	NN	O	O
12	NN	O	O
women	NN	O	O
with	NN	O	O
menopausal	NN	O	O
type	NN	O	O
II	NN	O	O
diabetes	NN	O	O
was	NN	O	O
measured	NN	O	O
with	NN	O	O
radio-ligand	NN	O	O
binding	NN	O	O
method	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
were	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
12	NN	O	O
menopausal	NN	O	O
women	NN	O	O
without	NN	O	O
diabetes	NN	O	O
and	NN	O	O
12	NN	O	O
normal	NN	O	O
women	NN	O	O
of	NN	O	O
childbearing	NN	O	O
age	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
in	NN	O	O
the	NN	O	O
patients	NN	O	O
was	NN	O	O
significantly	NN	O	O
decreased	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
decrease	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
level	NN	O	O
in	NN	O	O
leukocytes	NN	O	B-cell_type
may	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
type	NN	O	O
II	NN	O	O
diabetes	NN	O	O
in	NN	O	O
menopausal	NN	O	O
period	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lymphocyte	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
number	NN	O	O
in	NN	O	O
posttraumatic	NN	O	O
stress	NN	O	O
disorder	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
The	NN	O	O
authors	NN	O	O
'	NN	O	O
objective	NN	O	O
was	NN	O	O
to	NN	O	O
investigate	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
changes	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
dysregulation	NN	O	O
of	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
(	NN	O	O
HPA	NN	O	O
)	NN	O	O
axis	NN	O	O
in	NN	O	O
posttraumatic	NN	O	O
stress	NN	O	O
disorder	NN	O	O
(	NN	O	O
PTSD	NN	O	O
)	NN	O	O
.	NN	O	O

METHOD	NN	O	O
:	NN	O	O
They	NN	O	O
measured	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
lymphocyte	NN	O	B-protein
cytosolic	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
and	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
in	NN	O	O
15	NN	O	O
consecutively	NN	O	O
admitted	NN	O	O
male	NN	O	O
combat	NN	O	O
Vietnam	NN	O	O
veterans	NN	O	O
with	NN	O	O
PTSD	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
normal	NN	O	O
comparison	NN	O	O
group	NN	O	O
of	NN	O	O
11	NN	O	O
subjects	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Both	NN	O	O
the	NN	O	O
patients	NN	O	O
and	NN	O	O
the	NN	O	O
normal	NN	O	O
comparison	NN	O	O
subjects	NN	O	O
showed	NN	O	O
a	NN	O	O
morning-to-afternoon	NN	O	O
decline	NN	O	O
in	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentrations	NN	O	O
,	NN	O	O
paralleling	NN	O	O
the	NN	O	O
normal	NN	O	O
diurnal	NN	O	O
decline	NN	O	O
in	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
was	NN	O	O
63	NN	O	O
%	NN	O	O
greater	NN	O	O
in	NN	O	O
the	NN	O	O
morning	NN	O	O
and	NN	O	O
26	NN	O	O
%	NN	O	O
greater	NN	O	O
in	NN	O	O
the	NN	O	O
afternoon	NN	O	O
in	NN	O	O
the	NN	O	O
patients	NN	O	O
with	NN	O	O
PTSD	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
subjects	NN	O	O
.	NN	O	O

No	NN	O	O
group	NN	O	O
differences	NN	O	O
in	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
were	NN	O	O
observed	NN	O	O
,	NN	O	O
nor	NN	O	O
were	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
number	NN	O	O
and	NN	O	O
cortisol	NN	O	O
levels	NN	O	O
correlated	NN	O	O
.	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
morning	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
was	NN	O	O
positively	NN	O	O
correlated	NN	O	O
with	NN	O	O
symptoms	NN	O	O
of	NN	O	O
PTSD	NN	O	O
and	NN	O	O
anxiety	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
These	NN	O	O
results	NN	O	O
provide	NN	O	O
further	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
dysregulation	NN	O	O
of	NN	O	O
the	NN	O	O
HPA	NN	O	O
axis	NN	O	O
in	NN	O	O
PTSD	NN	O	O
.	NN	O	O

The	NN	O	O
finding	NN	O	O
that	NN	O	O
patients	NN	O	O
with	NN	O	O
PTSD	NN	O	O
had	NN	O	O
a	NN	O	O
substantially	NN	O	O
greater	NN	O	O
number	NN	O	O
of	NN	O	O
lymphocyte	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
than	NN	O	O
normal	NN	O	O
comparison	NN	O	O
subjects	NN	O	O
is	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
authors	NN	O	O
'	NN	O	O
previous	NN	O	O
observations	NN	O	O
of	NN	O	O
low	NN	O	O
24-hour	NN	O	O
urinary	NN	O	O
cortisol	NN	O	O
excretion	NN	O	O
in	NN	O	O
subjects	NN	O	O
with	NN	O	O
PTSD	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
receptor	NN	O	O
changes	NN	O	O
observed	NN	O	O
are	NN	O	O
opposite	NN	O	O
of	NN	O	O
those	NN	O	O
reported	NN	O	O
in	NN	O	O
major	NN	O	O
depressive	NN	O	O
disorder	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
data	NN	O	O
,	NN	O	O
along	NN	O	O
with	NN	O	O
other	NN	O	O
findings	NN	O	O
of	NN	O	O
HPA	NN	O	O
abnormalities	NN	O	O
in	NN	O	O
PTSD	NN	O	O
,	NN	O	O
support	NN	O	O
the	NN	O	O
possibility	NN	O	O
of	NN	O	O
a	NN	O	O
greater	NN	O	O
negative	NN	O	O
feedback	NN	O	O
sensitivity	NN	O	O
at	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
HPA	NN	O	O
axis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
during	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV-gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Cells	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte-macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
are	NN	O	O
important	NN	O	O
targets	NN	O	O
of	NN	O	O
HIV	NN	O	O
infection	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
the	NN	O	O
phenotypic	NN	O	O
differentiation	NN	O	O
of	NN	O	O
monocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
or	NN	O	O
tumour	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
expression	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
with	NN	O	O
such	NN	O	O
differentiation	NN	O	O
,	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
,	NN	O	O
monitored	NN	O	O
using	NN	O	O
an	NN	O	O
HIV	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
construct	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
activated	NN	O	O
in	NN	O	O
such	NN	O	O
cells	NN	O	O
under	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
enhanced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
expression	NN	O	O
.	NN	O	O

Also	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
promonocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
chronically	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV	NN	O	O
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
were	NN	O	O
enhanced	NN	O	O
on	NN	O	O
stimulation	NN	O	O
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
or	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
stimulation	NN	O	O
of	NN	O	O
monocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
or	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
induces	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
and	NN	O	O
activates	NN	O	O
HIV	NN	O	O
transcription	NN	O	O
.	NN	O	O

Such	NN	O	O
a	NN	O	O
process	NN	O	O
may	NN	O	O
have	NN	O	O
fundamental	NN	O	O
implications	NN	O	O
in	NN	O	O
AIDS	NN	O	O
pathogenesis	NN	O	O
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
important	NN	O	O
in	NN	O	O
disease	NN	O	O
progression	NN	O	O
induced	NN	O	O
by	NN	O	O
opportunistic	NN	O	O
infections	NN	O	O
directly	NN	O	O
or	NN	O	O
indirectly	NN	O	O
involving	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Demonstration	NN	O	O
of	NN	O	O
estrogen	NN	O	B-protein
and	NN	O	I-protein
progesterone	NN	O	I-protein
receptors	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
Ki-67	NN	O	B-protein
and	NN	O	I-protein
p-145	NN	O	I-protein
antigens	NN	O	I-protein
in	NN	O	O
single	NN	O	B-cell_type
tumor	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
blood	NN	O	O
and	NN	O	O
pleural	NN	O	O
effusions	NN	O	O
using	NN	O	O
a	NN	O	O
slide	NN	O	O
assay	NN	O	O
.	NN	O	O

We	NN	O	O
describe	NN	O	O
a	NN	O	O
slide	NN	O	O
assay	NN	O	O
that	NN	O	O
allows	NN	O	O
the	NN	O	O
demonstration	NN	O	O
of	NN	O	O
antigens	NN	O	B-protein
localized	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
from	NN	O	O
isolated	NN	O	B-cell_type
white	NN	O	I-cell_type
blood	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
from	NN	O	O
single	NN	O	B-cell_type
tumor	NN	O	I-cell_type
cells	NN	O	I-cell_type
derived	NN	O	O
from	NN	O	O
malignant	NN	O	O
effusions	NN	O	O
.	NN	O	O

With	NN	O	O
the	NN	O	O
antibodies	NN	O	O
Ki-67	NN	O	B-protein
and	NN	O	O
anti-p-145	NN	O	B-protein
an	NN	O	O
increased	NN	O	O
rate	NN	O	O
of	NN	O	O
nuclear	NN	O	O
and	NN	O	O
nucleolar	NN	O	O
staining	NN	O	O
resulted	NN	O	O
in	NN	O	O
cells	NN	O	O
from	NN	O	O
highly	NN	O	O
malignant	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

An	NN	O	O
almost	NN	O	O
identical	NN	O	O
reaction	NN	O	O
was	NN	O	O
obtained	NN	O	O
when	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
malignant	NN	O	O
effusions	NN	O	O
were	NN	O	O
tested	NN	O	O
.	NN	O	O

Cells	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
blood	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
leukemic	NN	O	O
spread	NN	O	O
of	NN	O	O
lymphomas	NN	O	O
of	NN	O	O
low	NN	O	O
malignancy	NN	O	O
yielded	NN	O	O
a	NN	O	O
weak	NN	O	O
staining	NN	O	O
comparable	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
mesothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
non-tumorous	NN	O	O
cavity	NN	O	O
fluids	NN	O	O
.	NN	O	O

The	NN	O	O
detection	NN	O	O
of	NN	O	O
estrogen	NN	O	B-protein
and	NN	O	O
progesterone	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
and	NN	O	O
PR	NN	O	B-protein
)	NN	O	O
localized	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
nucleus	NN	O	O
can	NN	O	O
be	NN	O	O
achieved	NN	O	O
by	NN	O	O
the	NN	O	O
same	NN	O	O
assay	NN	O	O
.	NN	O	O

The	NN	O	O
reaction	NN	O	O
is	NN	O	O
enhanced	NN	O	O
by	NN	O	O
incubation	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
30	NN	O	O
min	NN	O	O
at	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
prior	NN	O	O
to	NN	O	O
fixation	NN	O	O
.	NN	O	O

Pleural	NN	O	O
effusions	NN	O	O
from	NN	O	O
20	NN	O	O
patients	NN	O	O
with	NN	O	O
breast	NN	O	O
cancer	NN	O	O
were	NN	O	O
tested	NN	O	O
.	NN	O	O

ER	NN	O	B-protein
was	NN	O	O
positive	NN	O	O
in	NN	O	O
13	NN	O	O
and	NN	O	O
PR	NN	O	B-protein
was	NN	O	O
positive	NN	O	O
in	NN	O	O
12	NN	O	O
of	NN	O	O
the	NN	O	O
20	NN	O	O
samples	NN	O	O
.	NN	O	O

In	NN	O	O
5	NN	O	O
cases	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
divergent	NN	O	O
reaction	NN	O	O
with	NN	O	O
ER	NN	O	B-protein
and	NN	O	O
PR	NN	O	B-protein
antibody	NN	O	O
.	NN	O	O

The	NN	O	O
hormone	NN	O	B-protein
receptors	NN	O	I-protein
of	NN	O	O
the	NN	O	O
primary	NN	O	O
tumor	NN	O	O
were	NN	O	O
known	NN	O	O
in	NN	O	O
15	NN	O	O
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
and	NN	O	O
14	NN	O	O
(	NN	O	O
PR	NN	O	B-protein
)	NN	O	O
patients	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

In	NN	O	O
each	NN	O	O
cohort	NN	O	O
there	NN	O	O
was	NN	O	O
only	NN	O	O
one	NN	O	O
case	NN	O	O
with	NN	O	O
a	NN	O	O
negative	NN	O	O
reaction	NN	O	O
of	NN	O	O
the	NN	O	O
primary	NN	O	O
tumor	NN	O	O
and	NN	O	O
a	NN	O	O
positive	NN	O	O
reaction	NN	O	O
with	NN	O	O
the	NN	O	O
isolated	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
the	NN	O	O
pleural	NN	O	O
effusions	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
demonstration	NN	O	O
of	NN	O	O
hormone	NN	O	B-protein
receptor	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
cells	NN	O	O
from	NN	O	O
malignant	NN	O	O
effusions	NN	O	O
is	NN	O	O
possible	NN	O	O
and	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
correlation	NN	O	O
with	NN	O	O
the	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
primary	NN	O	O
site	NN	O	O
of	NN	O	O
cancer	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kB	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
thymocytes	NN	O	I-cell_type
.	NN	O	O

NF-kB	NN	O	B-protein
is	NN	O	O
a	NN	O	O
eukaryotic	NN	O	B-protein
transcription	NN	O	I-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
NF-kB	NN	O	B-protein
is	NN	O	O
bound	NN	O	O
to	NN	O	O
a	NN	O	O
cytoplasmic	NN	O	B-protein
proteic	NN	O	I-protein
inhibitor	NN	O	I-protein
,	NN	O	O
the	NN	O	O
IkB	NN	O	B-protein
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
mitogens	NN	O	O
(	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
)	NN	O	O
or	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
induces	NN	O	O
NF-kB	NN	O	B-protein
nuclear	NN	O	O
translocation	NN	O	O
and	NN	O	O
the	NN	O	O
subsequent	NN	O	O
expression	NN	O	O
of	NN	O	O
NF-kB	NN	O	B-DNA
dependent	NN	O	I-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kB	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
thymic	NN	O	I-cell_type
progenitors	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
report	NN	O	O
differences	NN	O	O
in	NN	O	O
(	NN	O	O
Ca2+	NN	O	O
)	NN	O	O
i	NN	O	O
requirement	NN	O	O
for	NN	O	O
NF-kB	NN	O	B-protein
activation	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
as	NN	O	O
compared	NN	O	O
to	NN	O	O
mature	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
indicated	NN	O	O
that	NN	O	O
thymocytes	NN	O	B-cell_type
have	NN	O	O
a	NN	O	O
constitutively	NN	O	B-protein
active	NN	O	I-protein
form	NN	O	I-protein
of	NN	O	O
NF-kB	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
they	NN	O	O
are	NN	O	O
activated	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

TCF-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
T	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
interacts	NN	O	O
with	NN	O	O
sequence	NN	O	B-DNA
motifs	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
beta	NN	O	I-DNA
and	NN	O	I-DNA
TCR	NN	O	I-DNA
delta	NN	O	I-DNA
enhancers	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
identified	NN	O	O
and	NN	O	O
cloned	NN	O	O
TCF-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
T	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
with	NN	O	O
specificity	NN	O	O
for	NN	O	O
the	NN	O	O
AACAAAG	NN	O	O
motif	NN	O	O
in	NN	O	O
the	NN	O	O
CD3	NN	O	O
epsilon	NN	O	O
enhancer	NN	O	O
and	NN	O	O
for	NN	O	O
the	NN	O	O
TTCAAAG	NN	O	B-DNA
motif	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
alpha	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

TCF-1	NN	O	B-protein
belongs	NN	O	O
to	NN	O	O
the	NN	O	O
family	NN	O	O
of	NN	O	O
transcription-regulating	NN	O	B-protein
proteins	NN	O	I-protein
which	NN	O	O
share	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
homology	NN	O	O
termed	NN	O	O
the	NN	O	O
HMG-box	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
by	NN	O	O
gel	NN	O	O
retardation	NN	O	O
analysis	NN	O	O
that	NN	O	O
TCF-1	NN	O	B-protein
specifically	NN	O	O
recognizes	NN	O	O
the	NN	O	O
T	NN	O	B-DNA
beta	NN	O	I-DNA
5	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
beta	NN	O	I-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
T	NN	O	B-DNA
delta	NN	O	I-DNA
7	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
delta	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Comparison	NN	O	O
of	NN	O	O
the	NN	O	O
sequences	NN	O	O
of	NN	O	O
all	NN	O	O
elements	NN	O	B-DNA
recognized	NN	O	O
by	NN	O	O
TCF-1	NN	O	B-protein
defines	NN	O	O
a	NN	O	O
consensus	NN	O	B-DNA
motif	NN	O	I-DNA
A/T	NN	O	B-DNA
A/T	NN	O	I-DNA
C	NN	O	I-DNA
A	NN	O	I-DNA
A/G	NN	O	I-DNA
A	NN	O	I-DNA
G	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
observations	NN	O	O
imply	NN	O	O
that	NN	O	O
TCF-1	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
several	NN	O	O
T	NN	O	B-DNA
cell-specific	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
might	NN	O	O
thus	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
establishment	NN	O	O
and	NN	O	O
maintenance	NN	O	O
of	NN	O	O
the	NN	O	O
mature	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
phenotype	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Regulatory	NN	O	O
effect	NN	O	O
of	NN	O	O
insulin	NN	O	B-protein
on	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
]	NN	O	O

The	NN	O	O
regulatory	NN	O	O
effect	NN	O	O
of	NN	O	O
insulin	NN	O	B-protein
on	NN	O	O
the	NN	O	O
specific	NN	O	O
binding	NN	O	O
power	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
was	NN	O	O
assessed	NN	O	O
by	NN	O	O
the	NN	O	O
unoccupied	NN	O	B-protein
receptor	NN	O	I-protein
sites	NN	O	I-protein
capable	NN	O	O
of	NN	O	O
combining	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
labelled	NN	O	O
dexamethasone	NN	O	O
measured	NN	O	O
at	NN	O	O
3	NN	O	O
and	NN	O	O
24	NN	O	O
h	NN	O	O
after	NN	O	O
incubation	NN	O	O
with	NN	O	O
various	NN	O	O
concentrations	NN	O	O
of	NN	O	O
insulin	NN	O	B-protein
added	NN	O	O
to	NN	O	O
the	NN	O	O
medium	NN	O	O
.	NN	O	O

After	NN	O	O
3	NN	O	O
h	NN	O	O
incubation	NN	O	O
the	NN	O	O
specific	NN	O	O
binding	NN	O	O
power	NN	O	O
with	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
Dex	NN	O	O
was	NN	O	O
decreased	NN	O	O
by	NN	O	O
23.3	NN	O	O
+/-	NN	O	O
10.0	NN	O	O
,	NN	O	O
32.2	NN	O	O
+/-	NN	O	O
13.2	NN	O	O
and	NN	O	O
54.3	NN	O	O
+/-	NN	O	O
9.2	NN	O	O
%	NN	O	O
(	NN	O	O
P	NN	O	O
greater	NN	O	O
than	NN	O	O
0.05	NN	O	O
,	NN	O	O
P	NN	O	O
greater	NN	O	O
than	NN	O	O
0.05	NN	O	O
and	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.01	NN	O	O
as	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
control	NN	O	O
value	NN	O	O
of	NN	O	O
100	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
insulin	NN	O	B-protein
)	NN	O	O
respectively	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
20	NN	O	O
mU/L	NN	O	O
(	NN	O	O
physiological	NN	O	O
testing	NN	O	O
concentration	NN	O	O
)	NN	O	O
,	NN	O	O
200	NN	O	O
mU/L	NN	O	O
(	NN	O	O
physiological	NN	O	O
upper	NN	O	O
limit	NN	O	O
)	NN	O	O
and	NN	O	O
2	NN	O	O
,	NN	O	O
000	NN	O	O
mU/L	NN	O	O
(	NN	O	O
pharmacological	NN	O	O
concentration	NN	O	O
)	NN	O	O
insulin	NN	O	B-protein
in	NN	O	O
the	NN	O	O
incubation	NN	O	O
medium	NN	O	O
.	NN	O	O

After	NN	O	O
24	NN	O	O
h	NN	O	O
incubation	NN	O	O
the	NN	O	O
decrease	NN	O	O
of	NN	O	O
these	NN	O	O
values	NN	O	O
increased	NN	O	O
respectively	NN	O	O
to	NN	O	O
43.5	NN	O	O
+/-	NN	O	O
19.0	NN	O	O
,	NN	O	O
56.1	NN	O	O
+/-	NN	O	O
20.7	NN	O	O
and	NN	O	O
80.2	NN	O	O
+/-	NN	O	O
15.5	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.05	NN	O	O
,	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.01	NN	O	O
and	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.01	NN	O	O
compared	NN	O	O
with	NN	O	O
control	NN	O	O
)	NN	O	O
.	NN	O	O

Thus	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
insulin	NN	O	B-protein
on	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
binding	NN	O	O
power	NN	O	O
is	NN	O	O
both	NN	O	O
dose-	NN	O	O
and	NN	O	O
time-dependent	NN	O	O
,	NN	O	O
which	NN	O	O
strongly	NN	O	O
suggests	NN	O	O
that	NN	O	O
GR	NN	O	B-protein
is	NN	O	O
tonically	NN	O	O
controlled	NN	O	O
by	NN	O	O
insulin	NN	O	B-protein
concentration	NN	O	O
change	NN	O	O
under	NN	O	O
physiological	NN	O	O
conditions	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
toremifene	NN	O	O
therapy	NN	O	O
on	NN	O	O
serum	NN	O	B-protein
immunoglobulin	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
.	NN	O	O

Estrogens	NN	O	O
and	NN	O	O
anti-estrogens	NN	O	O
enhance	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-cell_type
(	NN	O	I-cell_type
Ig	NN	O	I-cell_type
)	NN	O	I-cell_type
-secreting	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
pokeweed	NN	O	B-cell_line
mitogen	NN	O	I-cell_line
(	NN	O	I-cell_line
PWM	NN	O	I-cell_line
)	NN	O	I-cell_line
-stimulated	NN	O	I-cell_line
lymphocyte	NN	O	I-cell_line
cultures	NN	O	I-cell_line
.	NN	O	O

Lymphocytes	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
who	NN	O	O
have	NN	O	O
received	NN	O	O
anti-estrogen	NN	O	O
therapy	NN	O	O
show	NN	O	O
similar	NN	O	O
enhancement	NN	O	O
of	NN	O	O
Ig-secreting	NN	O	B-cell_type
cells	NN	O	I-cell_type
after	NN	O	O
PWM	NN	O	B-protein
stimulation	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
anti-estrogen	NN	O	O
(	NN	O	O
toremifene	NN	O	O
)	NN	O	O
therapy	NN	O	O
on	NN	O	O
serum	NN	O	B-protein
immunoglobulin	NN	O	I-protein
(	NN	O	O
IgA	NN	O	B-protein
,	NN	O	O
IgM	NN	O	B-protein
,	NN	O	O
IgG	NN	O	B-protein
)	NN	O	O
levels	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
was	NN	O	O
investigated	NN	O	O
.	NN	O	O

Serum	NN	O	O
Ig	NN	O	O
levels	NN	O	O
were	NN	O	O
followed	NN	O	O
up	NN	O	O
to	NN	O	O
two	NN	O	O
years	NN	O	O
after	NN	O	O
or	NN	O	O
during	NN	O	O
the	NN	O	O
therapy	NN	O	O
.	NN	O	O

An	NN	O	O
unexpected	NN	O	O
finding	NN	O	O
was	NN	O	O
that	NN	O	O
the	NN	O	O
Ig	NN	O	O
levels	NN	O	O
decreased	NN	O	O
during	NN	O	O
the	NN	O	O
follow-up	NN	O	O
period	NN	O	O
.	NN	O	O

This	NN	O	O
decrease	NN	O	O
was	NN	O	O
seen	NN	O	O
in	NN	O	O
patients	NN	O	O
who	NN	O	O
responded	NN	O	O
to	NN	O	O
the	NN	O	O
therapy	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
those	NN	O	O
who	NN	O	O
did	NN	O	O
not	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	B-protein
erythroid	NN	O	I-protein
5-aminolevulinate	NN	O	I-protein
synthase	NN	O	I-protein
:	NN	O	O
promoter	NN	O	B-DNA
analysis	NN	O	O
and	NN	O	O
identification	NN	O	O
of	NN	O	O
an	NN	O	O
iron-responsive	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
.	NN	O	O

5-Aminolevulinate	NN	O	B-protein
synthase	NN	O	I-protein
(	NN	O	O
ALAS	NN	O	B-protein
)	NN	O	O
catalyzes	NN	O	O
the	NN	O	O
first	NN	O	O
step	NN	O	O
of	NN	O	O
the	NN	O	O
heme	NN	O	B-protein
biosynthetic	NN	O	O
pathway	NN	O	O
.	NN	O	O

cDNA	NN	O	B-DNA
clones	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
human	NN	O	B-protein
erythroid	NN	O	I-protein
ALAS	NN	O	I-protein
isozyme	NN	O	I-protein
were	NN	O	O
isolated	NN	O	O
from	NN	O	O
a	NN	O	O
fetal	NN	O	O
liver	NN	O	O
library	NN	O	O
.	NN	O	O

It	NN	O	O
can	NN	O	O
be	NN	O	O
deduced	NN	O	O
that	NN	O	O
the	NN	O	O
erythroid	NN	O	B-protein
ALAS	NN	O	I-protein
precursor	NN	O	I-protein
protein	NN	O	I-protein
has	NN	O	O
a	NN	O	O
molecular	NN	O	O
weight	NN	O	O
of	NN	O	O
64.6	NN	O	O
kd	NN	O	O
,	NN	O	O
and	NN	O	O
is	NN	O	O
similar	NN	O	O
in	NN	O	O
size	NN	O	O
to	NN	O	O
the	NN	O	O
previously	NN	O	O
isolated	NN	O	O
human	NN	O	B-protein
housekeeping	NN	O	I-protein
ALAS	NN	O	I-protein
precursor	NN	O	I-protein
of	NN	O	O
molecular	NN	O	O
weight	NN	O	O
70.6	NN	O	O
kd	NN	O	O
.	NN	O	O

The	NN	O	O
mature	NN	O	O
mitochondrial	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
erythroid	NN	O	O
and	NN	O	O
housekeeping	NN	O	B-protein
ALAS	NN	O	I-protein
isozymes	NN	O	I-protein
are	NN	O	O
predicted	NN	O	O
to	NN	O	O
have	NN	O	O
molecular	NN	O	O
weights	NN	O	O
of	NN	O	O
59.5	NN	O	O
kd	NN	O	O
and	NN	O	O
64.6	NN	O	O
kd	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
isozymes	NN	O	B-protein
show	NN	O	O
little	NN	O	O
amino	NN	O	O
acid	NN	O	O
identity	NN	O	O
in	NN	O	O
their	NN	O	O
N-terminal	NN	O	B-protein
signal	NN	O	I-protein
sequences	NN	O	I-protein
but	NN	O	O
have	NN	O	O
considerable	NN	O	O
sequence	NN	O	O
identity	NN	O	O
in	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
two-thirds	NN	O	I-protein
of	NN	O	O
their	NN	O	O
proteins	NN	O	O
.	NN	O	O

An	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
immediate	NN	O	B-DNA
promoter	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
erythroid	NN	O	I-DNA
ALAS	NN	O	I-DNA
gene	NN	O	I-DNA
revealed	NN	O	O
several	NN	O	O
putative	NN	O	O
erythroid-specific	NN	O	B-DNA
cis-acting	NN	O	I-DNA
elements	NN	O	I-DNA
including	NN	O	O
both	NN	O	O
a	NN	O	O
GATA-1	NN	O	B-DNA
and	NN	O	O
an	NN	O	O
NF-E2	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

An	NN	O	O
iron-responsive	NN	O	B-RNA
element	NN	O	I-RNA
(	NN	O	I-RNA
IRE	NN	O	I-RNA
)	NN	O	I-RNA
motif	NN	O	I-RNA
has	NN	O	O
been	NN	O	O
identified	NN	O	O
in	NN	O	O
the	NN	O	O
5'-untranslated	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-RNA
erythroid	NN	O	I-RNA
ALAS	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
but	NN	O	O
is	NN	O	O
not	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
housekeeping	NN	O	B-RNA
ALAS	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Gel	NN	O	O
retardation	NN	O	O
experiments	NN	O	O
established	NN	O	O
that	NN	O	O
this	NN	O	O
IRE	NN	O	B-RNA
motif	NN	O	I-RNA
formed	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
-	NN	O	I-protein
RNA	NN	O	I-protein
complex	NN	O	I-protein
with	NN	O	O
cytosolic	NN	O	O
extracts	NN	O	O
from	NN	O	O
human	NN	O	B-cell_line
K562	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
this	NN	O	O
binding	NN	O	O
was	NN	O	O
strongly	NN	O	O
competed	NN	O	O
with	NN	O	O
IRE	NN	O	B-RNA
transcripts	NN	O	I-RNA
from	NN	O	O
ferritin	NN	O	B-RNA
or	NN	O	I-RNA
transferrin	NN	O	I-RNA
receptor	NN	O	I-RNA
mRNAs	NN	O	I-RNA
.	NN	O	O

A	NN	O	O
transcript	NN	O	O
of	NN	O	O
the	NN	O	O
ALAS	NN	O	B-DNA
IRE	NN	O	I-DNA
,	NN	O	O
mutated	NN	O	O
in	NN	O	O
the	NN	O	O
conserved	NN	O	O
loop	NN	O	O
of	NN	O	O
the	NN	O	O
IRE	NN	O	B-DNA
,	NN	O	O
did	NN	O	O
not	NN	O	O
readily	NN	O	O
form	NN	O	O
this	NN	O	O
protein	NN	O	B-protein
-	NN	O	I-protein
RNA	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
IRE	NN	O	B-RNA
motif	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
ALAS	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
functional	NN	O	O
and	NN	O	O
imply	NN	O	O
that	NN	O	O
translation	NN	O	O
of	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
cellular	NN	O	O
iron	NN	O	O
availability	NN	O	O
during	NN	O	O
erythropoiesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
jun	NN	O	O
and	NN	O	O
fos	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
by	NN	O	O
the	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
M-CSF	NN	O	B-protein
)	NN	O	O
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
signaling	NN	O	O
events	NN	O	O
responsible	NN	O	O
for	NN	O	O
these	NN	O	O
effects	NN	O	O
remain	NN	O	O
unclear	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
studies	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
M-CSF	NN	O	B-protein
on	NN	O	O
potential	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
involving	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
jun	NN	O	B-DNA
and	NN	O	I-DNA
fos	NN	O	I-DNA
early	NN	O	I-DNA
response	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Low	NN	O	O
levels	NN	O	O
of	NN	O	O
c-jun	NN	O	B-protein
transcripts	NN	O	I-protein
were	NN	O	O
detectable	NN	O	O
in	NN	O	O
resting	NN	O	B-cell_type
human	NN	O	I-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
with	NN	O	O
10	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
units/ml	NN	O	O
human	NN	O	B-protein
recombinant	NN	O	I-protein
M-CSF	NN	O	I-protein
was	NN	O	O
associated	NN	O	O
with	NN	O	O
rapid	NN	O	O
and	NN	O	O
transient	NN	O	O
increases	NN	O	O
in	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
run-on	NN	O	O
assays	NN	O	O
and	NN	O	O
mRNA	NN	O	O
stability	NN	O	O
studies	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
M-CSF	NN	O	B-protein
regulates	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
by	NN	O	O
both	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
transcription	NN	O	O
rate	NN	O	O
and	NN	O	O
a	NN	O	O
prolongation	NN	O	O
in	NN	O	O
the	NN	O	O
half-life	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
transcripts	NN	O	I-RNA
.	NN	O	O

M-CSF	NN	O	B-protein
treatment	NN	O	O
was	NN	O	O
also	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
rapid	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
jun-B	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
although	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
was	NN	O	O
prolonged	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
M-CSF	NN	O	B-protein
increases	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
through	NN	O	O
control	NN	O	O
at	NN	O	O
both	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
posttranscriptional	NN	O	O
levels	NN	O	O
.	NN	O	O

Maximal	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
c-fos	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
followed	NN	O	O
by	NN	O	O
that	NN	O	O
for	NN	O	O
the	NN	O	O
fos-B	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
M-CSF	NN	O	B-protein
-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
fos-related	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
fra-1	NN	O	B-DNA
,	NN	O	O
was	NN	O	O
delayed	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
for	NN	O	O
both	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
fos-B	NN	O	B-DNA
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
the	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
M-CSF	NN	O	B-protein
treatment	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
differential	NN	O	O
activation	NN	O	O
of	NN	O	O
multiple	NN	O	O
members	NN	O	O
of	NN	O	O
the	NN	O	O
jun/fos	NN	O	B-DNA
family	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
could	NN	O	O
contribute	NN	O	O
to	NN	O	O
nuclear	NN	O	O
signaling	NN	O	O
mechanisms	NN	O	O
that	NN	O	O
regulate	NN	O	O
a	NN	O	O
specific	NN	O	O
program	NN	O	O
of	NN	O	O
monocyte	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
.	NN	O	O

Glucocorticosteroids	NN	O	O
remain	NN	O	O
the	NN	O	O
major	NN	O	O
treatment	NN	O	O
modality	NN	O	O
for	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
(	NN	O	O
SLE	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
their	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
is	NN	O	O
unclear	NN	O	O
.	NN	O	O

Over	NN	O	O
the	NN	O	O
past	NN	O	O
decade	NN	O	O
it	NN	O	O
has	NN	O	O
become	NN	O	O
clear	NN	O	O
that	NN	O	O
glucocorticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
play	NN	O	O
a	NN	O	O
significant	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
glucocorticosteroid	NN	O	O
action	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
glucocorticosteroid	NN	O	B-protein
receptor	NN	O	I-protein
density	NN	O	O
and	NN	O	O
affinity	NN	O	O
on	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
the	NN	O	O
glucocorticosteroid	NN	O	O
binding	NN	O	O
assay	NN	O	O
in	NN	O	O
33	NN	O	O
patients	NN	O	O
with	NN	O	O
SLE	NN	O	O
who	NN	O	O
had	NN	O	O
taken	NN	O	O
no	NN	O	O
glucocorticosteroid	NN	O	O
for	NN	O	O
the	NN	O	O
previous	NN	O	O
6	NN	O	O
months	NN	O	O
and	NN	O	O
in	NN	O	O
32	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

Patients	NN	O	O
'	NN	O	O
disease	NN	O	O
activity	NN	O	O
was	NN	O	O
measured	NN	O	O
by	NN	O	O
the	NN	O	O
SLE	NN	O	O
Disease	NN	O	O
Activity	NN	O	O
Index	NN	O	O
(	NN	O	O
SLEDAI	NN	O	O
)	NN	O	O
.	NN	O	O

Glucocorticosteroid	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
leukocytes	NN	O	B-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
SLE	NN	O	O
were	NN	O	O
significantly	NN	O	O
higher	NN	O	O
than	NN	O	O
in	NN	O	O
healthy	NN	O	O
controls	NN	O	O
(	NN	O	O
4419	NN	O	O
+/-	NN	O	O
306	NN	O	O
vs	NN	O	O
3369	NN	O	O
+/-	NN	O	O
196	NN	O	O
,	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0.005	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
binding	NN	O	O
affinity	NN	O	O
was	NN	O	O
not	NN	O	O
different	NN	O	O
between	NN	O	O
patients	NN	O	O
and	NN	O	O
controls	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
correlation	NN	O	O
between	NN	O	O
glucocorticosteroid	NN	O	B-protein
receptor	NN	O	I-protein
number	NN	O	O
and	NN	O	O
SLE	NN	O	O
disease	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
an	NN	O	O
IL-2	NN	O	B-cell_line
autocrine	NN	O	I-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
IL-2	NN	O	B-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
IL-2-R	NN	O	I-DNA
alpha	NN	O	I-DNA
genes	NN	O	I-DNA
are	NN	O	O
both	NN	O	O
expressed	NN	O	O
transiently	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
after	NN	O	O
Ag	NN	O	O
or	NN	O	O
mitogen	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
IARC	NN	O	B-cell_line
301	NN	O	I-cell_line
,	NN	O	O
expresses	NN	O	O
these	NN	O	O
two	NN	O	O
genes	NN	O	O
constitutively	NN	O	O
and	NN	O	O
we	NN	O	O
have	NN	O	O
previously	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
its	NN	O	O
growth	NN	O	O
depends	NN	O	O
on	NN	O	O
the	NN	O	O
autocrine	NN	O	O
production	NN	O	O
of	NN	O	O
this	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
high	NN	O	B-protein
affinity	NN	O	I-protein
IL-2R	NN	O	I-protein
.	NN	O	O

To	NN	O	O
dissect	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
for	NN	O	O
the	NN	O	O
unusual	NN	O	O
persistent	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
and	NN	O	I-DNA
IL-2-R	NN	O	I-DNA
alpha	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
these	NN	O	O
IARC	NN	O	B-cell_line
301	NN	O	I-cell_line
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
interactions	NN	O	O
of	NN	O	O
constitutively	NN	O	B-protein
expressed	NN	O	I-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
with	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
and	NN	O	I-DNA
IL-2-R	NN	O	I-DNA
alpha	NN	O	I-DNA
genes	NN	O	I-DNA
using	NN	O	O
both	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
footprinting	NN	O	O
and	NN	O	O
gel	NN	O	O
retardation	NN	O	O
techniques	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
a	NN	O	O
region	NN	O	O
in	NN	O	O
both	NN	O	O
genes	NN	O	O
(	NN	O	O
-276	NN	O	O
to	NN	O	O
-250	NN	O	O
for	NN	O	O
IL-2-R	NN	O	B-DNA
alpha	NN	O	I-DNA
and	NN	O	O
-203	NN	O	O
to	NN	O	O
-183	NN	O	O
for	NN	O	O
IL-2	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
which	NN	O	O
corresponds	NN	O	O
to	NN	O	O
a	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	I-DNA
is	NN	O	O
specifically	NN	O	O
protected	NN	O	O
by	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
from	NN	O	O
IARC	NN	O	B-cell_line
301	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
agreement	NN	O	O
with	NN	O	O
this	NN	O	O
finding	NN	O	O
,	NN	O	O
both	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
and	NN	O	I-DNA
IL-2-R	NN	O	I-DNA
alpha	NN	O	I-DNA
promoters	NN	O	I-DNA
are	NN	O	O
active	NN	O	O
in	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
in	NN	O	O
IARC	NN	O	B-cell_line
301	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
kappa	NN	O	B-protein
B	NN	O	I-protein
enhancer	NN	O	I-protein
results	NN	O	O
in	NN	O	O
markedly	NN	O	O
attenuated	NN	O	O
activities	NN	O	O
of	NN	O	O
both	NN	O	O
promoters	NN	O	B-DNA
.	NN	O	O

Two	NN	O	O
proteins	NN	O	O
binding	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sequence	NN	O	I-DNA
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
KBF1	NN	O	B-protein
,	NN	O	O
are	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
IARC	NN	O	B-cell_line
301	NN	O	I-cell_line
nuclei	NN	O	O
and	NN	O	O
induced	NN	O	O
by	NN	O	O
PMA	NN	O	O
and	NN	O	O
PHA	NN	O	B-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
.	NN	O	O

They	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
motifs	NN	O	I-DNA
with	NN	O	O
different	NN	O	O
relative	NN	O	O
affinities	NN	O	O
that	NN	O	O
may	NN	O	O
reflect	NN	O	O
their	NN	O	O
different	NN	O	O
contribution	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
various	NN	O	O
promoters	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

HIV1	NN	O	O
infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
promotes	NN	O	O
induction	NN	O	O
or	NN	O	O
translocation	NN	O	O
of	NN	O	O
NF-KB-related	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
1991	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrated	NN	O	O
,	NN	O	O
using	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
,	NN	O	O
that	NN	O	O
3	NN	O	O
different	NN	O	O
factors	NN	O	O
(	NN	O	O
termed	NN	O	O
B1	NN	O	B-protein
,	NN	O	O
B2	NN	O	B-protein
and	NN	O	O
B3	NN	O	B-protein
)	NN	O	O
with	NN	O	O
affinity	NN	O	O
for	NN	O	O
the	NN	O	O
KB-enhancer	NN	O	B-DNA
target	NN	O	I-DNA
sequence	NN	O	I-DNA
were	NN	O	O
specifically	NN	O	O
detected	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
HIV1-infected	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
B2	NN	O	B-protein
factor	NN	O	I-protein
was	NN	O	O
induced	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
only	NN	O	O
upon	NN	O	O
HIV1	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
B3	NN	O	B-protein
factor	NN	O	I-protein
was	NN	O	O
only	NN	O	O
slightly	NN	O	O
evident	NN	O	O
in	NN	O	O
nuclei	NN	O	O
of	NN	O	O
uninfected	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
was	NN	O	O
readily	NN	O	O
detectable	NN	O	O
in	NN	O	O
nuclei	NN	O	O
of	NN	O	O
infected	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Its	NN	O	O
expression	NN	O	O
remained	NN	O	O
very	NN	O	O
low	NN	O	O
in	NN	O	O
nuclei	NN	O	O
of	NN	O	O
HIV1-infected	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
paper	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
B2	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
monocytes	NN	O	B-cell_type
and	NN	O	O
macrophages	NN	O	B-cell_type
as	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
(	NN	O	O
IKB	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
factor	NN	O	O
remained	NN	O	O
clustered	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
and	NN	O	O
was	NN	O	O
translocated	NN	O	O
to	NN	O	O
the	NN	O	O
nuclei	NN	O	O
only	NN	O	O
after	NN	O	O
HIV1	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
B3	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
only	NN	O	O
when	NN	O	O
cells	NN	O	O
are	NN	O	O
HIV1-infected	NN	O	O
.	NN	O	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
HIV1	NN	O	O
infection	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
the	NN	O	O
translocation	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
is	NN	O	O
discussed	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
and	NN	O	O
cloning	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
T	NN	O	B-protein
lymphocyte-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
containing	NN	O	O
a	NN	O	O
sequence-specific	NN	O	B-protein
HMG	NN	O	I-protein
box	NN	O	I-protein
.	NN	O	O

CD3	NN	O	B-protein
-epsilon	NN	O	O
expression	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
downstream	NN	O	B-DNA
T	NN	O	I-DNA
lymphocyte-specific	NN	O	I-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
report	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
a	NN	O	O
T	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
TCF-1	NN	O	B-protein
,	NN	O	O
binding	NN	O	O
to	NN	O	O
this	NN	O	O
element	NN	O	O
.	NN	O	O

The	NN	O	O
multimerized	NN	O	B-protein
recognition	NN	O	I-protein
motif	NN	O	I-protein
of	NN	O	O
TCF-1	NN	O	B-protein
constituted	NN	O	O
a	NN	O	O
T	NN	O	O
cell-specific	NN	O	O
enhancer	NN	O	O
.	NN	O	O

Subsequent	NN	O	O
cloning	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
identified	NN	O	O
three	NN	O	O
splice	NN	O	B-protein
alternatives	NN	O	I-protein
.	NN	O	O

TCF-1	NN	O	B-protein
contained	NN	O	O
a	NN	O	O
single	NN	O	O
DNA-binding	NN	O	B-protein
HMG	NN	O	I-protein
box	NN	O	I-protein
most	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
similar	NN	O	O
boxes	NN	O	O
in	NN	O	O
the	NN	O	O
putative	NN	O	B-DNA
mammalian	NN	O	I-DNA
sex-determining	NN	O	I-DNA
gene	NN	O	I-DNA
SRY	NN	O	B-DNA
and	NN	O	O
in	NN	O	O
the	NN	O	O
Schizosaccharomyces	NN	O	B-DNA
pombe	NN	O	I-DNA
Mc	NN	O	I-DNA
mating	NN	O	I-DNA
type	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

TCF-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
expressed	NN	O	O
uniquely	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Upon	NN	O	O
cotransfection	NN	O	O
into	NN	O	O
non-T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
TCF-1	NN	O	B-protein
could	NN	O	O
transactivate	NN	O	O
through	NN	O	O
its	NN	O	O
cognate	NN	O	O
motif	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
identify	NN	O	O
TCF-1	NN	O	B-protein
as	NN	O	O
a	NN	O	O
T	NN	O	B-protein
cell-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
which	NN	O	O
might	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
establishment	NN	O	O
of	NN	O	O
the	NN	O	O
mature	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
phenotype	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
suppressor	NN	O	I-protein
proteins	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
.	NN	O	O

Two	NN	O	O
different	NN	O	O
proteins	NN	O	O
which	NN	O	O
independently	NN	O	O
bound	NN	O	O
to	NN	O	O
neighboring	NN	O	O
sequences	NN	O	O
within	NN	O	O
the	NN	O	O
negative	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
NRE	NN	O	B-DNA
)	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
extract	NN	O	O
of	NN	O	O
a	NN	O	O
virus-infected	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
factors	NN	O	O
bound	NN	O	O
to	NN	O	O
a	NN	O	O
novel	NN	O	B-DNA
dyad	NN	O	I-DNA
symmetrical	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
sequence	NN	O	O
is	NN	O	O
well	NN	O	O
conserved	NN	O	O
in	NN	O	O
various	NN	O	O
HIV-1	NN	O	O
isolates	NN	O	O
and	NN	O	O
partial	NN	O	O
homology	NN	O	O
was	NN	O	O
found	NN	O	O
with	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
retinoblastoma	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Similar	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
virus-uninfected	NN	O	B-cell_line
human	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
means	NN	O	O
of	NN	O	O
a	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
factor	NN	O	O
bound	NN	O	O
to	NN	O	O
a	NN	O	O
putative	NN	O	B-DNA
AP-1	NN	O	I-DNA
recognition	NN	O	I-DNA
sequence	NN	O	I-DNA
predicted	NN	O	O
for	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
NRE	NN	O	B-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
this	NN	O	O
factor	NN	O	O
did	NN	O	O
not	NN	O	O
bind	NN	O	O
to	NN	O	O
a	NN	O	O
typical	NN	O	O
AP-1	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
insertion	NN	O	O
of	NN	O	O
multiple	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
former	NN	O	O
or	NN	O	O
latter	NN	O	O
factor	NN	O	O
into	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
reduced	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
to	NN	O	O
one-tenth	NN	O	O
or	NN	O	O
one-third	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
each	NN	O	O
factor	NN	O	O
may	NN	O	O
function	NN	O	O
as	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
negative	NN	O	I-protein
regulator	NN	O	I-protein
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

Protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
activation	NN	O	O
and	NN	O	O
protooncogene	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
differentiation/retrodifferentiation	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
U-937	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Human	NN	O	B-cell_line
U-937	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
differentiate	NN	O	O
along	NN	O	O
the	NN	O	O
monocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
following	NN	O	O
3-day	NN	O	O
exposures	NN	O	O
to	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
induction	NN	O	O
of	NN	O	O
differentiation	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
adherence	NN	O	O
and	NN	O	O
loss	NN	O	O
of	NN	O	O
proliferation	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
expression/repression	NN	O	O
of	NN	O	O
differentiation-associated	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Long	NN	O	O
term	NN	O	O
culture	NN	O	O
of	NN	O	O
TPA-differentiated	NN	O	B-cell_type
U-937	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
for	NN	O	O
32-36	NN	O	O
days	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
process	NN	O	O
of	NN	O	O
retrodifferentiation	NN	O	O
.	NN	O	O

The	NN	O	O
retrodifferentiated	NN	O	O
cells	NN	O	O
detached	NN	O	O
from	NN	O	O
the	NN	O	O
substrate	NN	O	O
and	NN	O	O
reinitiated	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Other	NN	O	O
cellular	NN	O	O
parameters	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
glycosidase	NN	O	O
activities	NN	O	O
,	NN	O	O
cytokine	NN	O	B-protein
release	NN	O	O
,	NN	O	O
and	NN	O	O
filament	NN	O	O
expression	NN	O	O
,	NN	O	O
returned	NN	O	O
to	NN	O	O
levels	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
observed	NN	O	O
in	NN	O	O
uninduced	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
TPA	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
rapid	NN	O	O
translocation	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
from	NN	O	O
the	NN	O	O
cytosol	NN	O	O
to	NN	O	O
cell	NN	O	O
membrane	NN	O	O
fractions	NN	O	O
within	NN	O	O
2-8	NN	O	O
min	NN	O	O
.	NN	O	O

Increased	NN	O	O
levels	NN	O	O
of	NN	O	O
membrane-associated	NN	O	O
PKC	NN	O	B-protein
activity	NN	O	O
persisted	NN	O	O
until	NN	O	O
17-29	NN	O	O
days	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
longer	NN	O	O
periods	NN	O	O
of	NN	O	O
incubation	NN	O	O
were	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
return	NN	O	O
to	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
in	NN	O	O
control	NN	O	O
cells	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
certain	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
response	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
present	NN	O	O
studies	NN	O	O
,	NN	O	O
TPA	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	I-DNA
.	NN	O	O

Levels	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
and	NN	O	I-RNA
c-jun	NN	O	I-RNA
transcripts	NN	O	I-RNA
remained	NN	O	O
elevated	NN	O	O
during	NN	O	O
periods	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
also	NN	O	O
returned	NN	O	O
to	NN	O	O
levels	NN	O	O
observed	NN	O	O
in	NN	O	O
control	NN	O	O
cells	NN	O	O
by	NN	O	O
30-36	NN	O	O
days	NN	O	O
,	NN	O	O
when	NN	O	O
the	NN	O	O
cells	NN	O	O
entered	NN	O	O
retrodifferentiation	NN	O	O
.	NN	O	O

Staurosporine	NN	O	O
,	NN	O	O
a	NN	O	O
nonspecific	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
PKC	NN	O	B-protein
,	NN	O	O
partially	NN	O	O
blocked	NN	O	O
TPA-induced	NN	O	O
adherence	NN	O	O
and	NN	O	O
growth	NN	O	O
inhibition	NN	O	O
and	NN	O	O
concomitantly	NN	O	O
prevented	NN	O	O
TPA-induced	NN	O	O
c-fos	NN	O	O
and	NN	O	O
c-jun	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

(	NN	O	O
ABSTRACT	NN	O	O
TRUNCATED	NN	O	O
AT	NN	O	O
250	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

Kappa	NN	O	B-protein
B-specific	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
differentially	NN	O	O
inhibited	NN	O	O
by	NN	O	O
enhancer	NN	O	O
mutations	NN	O	O
and	NN	O	O
biological	NN	O	O
oxidation	NN	O	O
.	NN	O	O

Kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
enhancer	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
nuclei	NN	O	O
of	NN	O	O
activated	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
produce	NN	O	O
two	NN	O	O
distinct	NN	O	O
nucleoprotein	NN	O	B-protein
complexes	NN	O	I-protein
when	NN	O	O
incubated	NN	O	O
with	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
element	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
receptor-alpha	NN	O	I-DNA
(	NN	O	I-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
two	NN	O	O
DNA-protein	NN	O	B-protein
complexes	NN	O	I-protein
are	NN	O	O
composed	NN	O	O
of	NN	O	O
at	NN	O	O
least	NN	O	O
four	NN	O	O
host	NN	O	O
proteins	NN	O	O
(	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p55	NN	O	B-protein
,	NN	O	O
p75	NN	O	B-protein
,	NN	O	O
p85	NN	O	B-protein
)	NN	O	O
,	NN	O	O
each	NN	O	O
of	NN	O	O
which	NN	O	O
shares	NN	O	O
structural	NN	O	O
similarity	NN	O	O
with	NN	O	O
the	NN	O	O
v-rel	NN	O	B-protein
oncogene	NN	O	I-protein
product	NN	O	I-protein
.	NN	O	O

Nuclear	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
is	NN	O	O
induced	NN	O	O
with	NN	O	O
distinctly	NN	O	O
biphasic	NN	O	O
kinetics	NN	O	O
following	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
p55/p75	NN	O	B-protein
early	NN	O	I-protein
and	NN	O	O
p50/p85	NN	O	B-protein
late	NN	O	I-protein
)	NN	O	O
.	NN	O	O

DNA-protein	NN	O	O
crosslinking	NN	O	O
studies	NN	O	O
have	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
more	NN	O	O
rapidly	NN	O	O
migrating	NN	O	O
B2	NN	O	B-protein
complex	NN	O	I-protein
contains	NN	O	O
both	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p55	NN	O	B-protein
while	NN	O	O
the	NN	O	O
more	NN	O	O
slowly	NN	O	O
migrating	NN	O	O
B1	NN	O	B-protein
complex	NN	O	I-protein
is	NN	O	O
composed	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
p55	NN	O	B-protein
,	NN	O	O
p75	NN	O	B-protein
,	NN	O	O
and	NN	O	O
p85	NN	O	B-protein
.	NN	O	O

Site-directed	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
wild-type	NN	O	B-DNA
IL-2R	NN	O	I-DNA
alpha	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
(	NN	O	O
GGGGAATCTCCC	NN	O	O
)	NN	O	O
has	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
p50	NN	O	O
and	NN	O	O
p55	NN	O	B-protein
(	NN	O	O
B2	NN	O	B-protein
complex	NN	O	I-protein
)	NN	O	O
is	NN	O	O
particularly	NN	O	O
sensitive	NN	O	O
to	NN	O	O
alteration	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
triplet	NN	O	I-DNA
of	NN	O	O
deoxyguanosine	NN	O	O
residues	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
B1	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
reflecting	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
p75	NN	O	B-protein
and	NN	O	O
p85	NN	O	B-protein
,	NN	O	O
critically	NN	O	O
depends	NN	O	O
upon	NN	O	O
the	NN	O	O
more	NN	O	O
3	NN	O	O
'	NN	O	O
sequences	NN	O	O
of	NN	O	O
this	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
all	NN	O	O
four	NN	O	O
of	NN	O	O
these	NN	O	O
Rel-related	NN	O	B-protein
factors	NN	O	I-protein
is	NN	O	O
blocked	NN	O	O
by	NN	O	O
selective	NN	O	O
chemical	NN	O	O
modification	NN	O	O
of	NN	O	O
lysine	NN	O	O
and	NN	O	O
arginine	NN	O	O
residues	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
both	NN	O	O
of	NN	O	O
these	NN	O	O
basic	NN	O	O
amino	NN	O	O
acids	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
covalent	NN	O	O
modification	NN	O	O
of	NN	O	O
free	NN	O	O
sulfhydryl	NN	O	O
groups	NN	O	O
with	NN	O	O
diamide	NN	O	O
(	NN	O	O
reversible	NN	O	O
)	NN	O	O
or	NN	O	O
N-ethylmaleimide	NN	O	O
(	NN	O	O
irreversible	NN	O	O
)	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
complete	NN	O	O
loss	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
mild	NN	O	O
oxidation	NN	O	O
with	NN	O	O
glucose	NN	O	B-protein
oxidase	NN	O	I-protein
selectively	NN	O	O
inhibits	NN	O	O
p75	NN	O	B-protein
and	NN	O	O
p85	NN	O	B-protein
binding	NN	O	O
while	NN	O	O
not	NN	O	O
blocking	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p55	NN	O	B-protein
interactions	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
reduced	NN	O	O
cysteine	NN	O	O
thiols	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
family	NN	O	O
of	NN	O	O
Rel-related	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Changes	NN	O	O
in	NN	O	O
leucocytic	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
climacteric	NN	O	O
syndrome	NN	O	O
and	NN	O	O
therapeutic	NN	O	O
effect	NN	O	O
of	NN	O	O
liuwei	NN	O	O
dihuang	NN	O	O
pills	NN	O	O
]	NN	O	O

The	NN	O	O
numbers	NN	O	O
of	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
in	NN	O	O
22	NN	O	O
women	NN	O	O
with	NN	O	O
climacteric	NN	O	O
syndrome	NN	O	O
were	NN	O	O
measured	NN	O	O
by	NN	O	O
radioligand	NN	O	O
method	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
were	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
12	NN	O	O
normal	NN	O	O
child-bearing-age	NN	O	O
women	NN	O	O
.	NN	O	O

It	NN	O	O
wat	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
contents	NN	O	O
of	NN	O	O
leucocytic	NN	O	B-protein
ER	NN	O	I-protein
in	NN	O	O
climacteric	NN	O	O
syndrome	NN	O	O
patients	NN	O	O
were	NN	O	O
significantly	NN	O	O
lower	NN	O	O
than	NN	O	O
normal	NN	O	O
child-bearing-age	NN	O	O
women	NN	O	O
.	NN	O	O

The	NN	O	O
authors	NN	O	O
used	NN	O	O
a	NN	O	O
Chinese	NN	O	O
prescription	NN	O	O
--	NN	O	O
Liuwei	NN	O	O
Dihuang	NN	O	O
Pills	NN	O	O
(	NN	O	O
LDP	NN	O	O
)	NN	O	O
to	NN	O	O
treat	NN	O	O
the	NN	O	O
patients	NN	O	O
for	NN	O	O
2	NN	O	O
months	NN	O	O
.	NN	O	O

The	NN	O	O
numbers	NN	O	O
of	NN	O	O
leucocytic	NN	O	B-protein
ER	NN	O	I-protein
were	NN	O	O
significantly	NN	O	O
increased	NN	O	O
after	NN	O	O
treatment	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
decrease	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
levels	NN	O	O
in	NN	O	O
cell	NN	O	O
may	NN	O	O
involve	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
climacteric	NN	O	O
syndrome	NN	O	O
.	NN	O	O

LDP	NN	O	O
not	NN	O	O
only	NN	O	O
increases	NN	O	O
plasma	NN	O	O
estradiol	NN	O	O
levels	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
increases	NN	O	O
the	NN	O	O
leucocytic	NN	O	O
ER	NN	O	B-protein
levels	NN	O	O
.	NN	O	O

This	NN	O	O
may	NN	O	O
be	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
therapeutic	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
disease	NN	O	O
.	NN	O	O

-DOCSTART-	O

Evaluation	NN	O	O
of	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
ligand	NN	O	O
and	NN	O	O
thermal	NN	O	O
activation	NN	O	O
of	NN	O	O
specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
in	NN	O	B-protein
vitro	NN	O	I-protein
synthesized	NN	O	I-protein
human	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
a	NN	O	O
DNA-binding/immunoprecipitation	NN	O	O
assay	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
capacity	NN	O	O
of	NN	O	O
human	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
hGR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
generated	NN	O	O
in	NN	O	O
rabbit	NN	O	O
reticulocyte	NN	O	O
lysates	NN	O	O
,	NN	O	O
to	NN	O	O
bind	NN	O	O
DNA	NN	O	O
.	NN	O	O

In	NN	O	B-protein
vitro	NN	O	I-protein
translated	NN	O	I-protein
hGR	NN	O	I-protein
was	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
native	NN	O	B-protein
hGR	NN	O	I-protein
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
migration	NN	O	O
on	NN	O	O
sodium	NN	O	O
dodecyl	NN	O	O
sulfate-polyacrylamide	NN	O	O
gels	NN	O	O
,	NN	O	O
sedimentation	NN	O	O
on	NN	O	O
sucrose	NN	O	O
density	NN	O	O
gradients	NN	O	O
,	NN	O	O
and	NN	O	O
reactivity	NN	O	O
with	NN	O	O
antipeptide	NN	O	B-protein
antibodies	NN	O	I-protein
generated	NN	O	O
against	NN	O	O
hGR	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
cell-free	NN	O	B-protein
synthesized	NN	O	I-protein
hGR	NN	O	I-protein
was	NN	O	O
capable	NN	O	O
of	NN	O	O
specific	NN	O	O
binding	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	I-DNA
GRE	NN	O	I-DNA
)	NN	O	I-DNA
-containing	NN	O	I-DNA
DNA	NN	O	I-DNA
fragments	NN	O	I-DNA
.	NN	O	O

Using	NN	O	O
this	NN	O	O
assay	NN	O	O
system	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
evaluated	NN	O	O
the	NN	O	O
contributions	NN	O	O
of	NN	O	O
ligand	NN	O	O
binding	NN	O	O
and	NN	O	O
heat	NN	O	O
activation	NN	O	O
to	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
these	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

In	NN	O	B-protein
vitro	NN	O	I-protein
translated	NN	O	I-protein
hGR	NN	O	I-protein
was	NN	O	O
capable	NN	O	O
of	NN	O	O
selective	NN	O	O
DNA	NN	O	O
binding	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
.	NN	O	O

Treatment	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
or	NN	O	O
the	NN	O	O
antiglucocorticoid	NN	O	O
RU486	NN	O	O
had	NN	O	O
no	NN	O	O
additional	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
DNA-binding	NN	O	O
capacity	NN	O	O
when	NN	O	O
receptor	NN	O	O
preparations	NN	O	O
were	NN	O	O
maintained	NN	O	O
at	NN	O	O
0	NN	O	O
C	NN	O	O
(	NN	O	O
no	NN	O	O
activation	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
addition	NN	O	O
of	NN	O	O
either	NN	O	O
ligand	NN	O	O
or	NN	O	O
antagonist	NN	O	O
in	NN	O	O
combination	NN	O	O
with	NN	O	O
a	NN	O	O
heat	NN	O	O
activation	NN	O	O
step	NN	O	O
promoted	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
approximately	NN	O	O
3-fold	NN	O	O
over	NN	O	O
that	NN	O	O
of	NN	O	O
heat-activated	NN	O	O
unliganded	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Agonist	NN	O	O
(	NN	O	O
dexamethasone	NN	O	O
)	NN	O	O
was	NN	O	O
slightly	NN	O	O
more	NN	O	O
effective	NN	O	O
in	NN	O	O
supporting	NN	O	O
specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
than	NN	O	O
antagonist	NN	O	O
(	NN	O	O
RU486	NN	O	O
)	NN	O	O
.	NN	O	O

DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
in	NN	O	B-protein
vitro	NN	O	I-protein
synthesized	NN	O	I-protein
GR	NN	O	I-protein
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
sodium	NN	O	O
molybdate	NN	O	O
to	NN	O	O
the	NN	O	O
receptor	NN	O	O
preparations	NN	O	O
before	NN	O	O
steroid	NN	O	O
addition	NN	O	O
and	NN	O	O
thermal	NN	O	O
activation	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
KCl	NN	O	O
resulted	NN	O	O
in	NN	O	O
less	NN	O	O
DNA	NN	O	O
binding	NN	O	O
either	NN	O	O
due	NN	O	O
to	NN	O	O
blockage	NN	O	O
of	NN	O	O
DNA-	NN	O	O
receptor	NN	O	B-protein
complex	NN	O	I-protein
formation	NN	O	O
or	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
complexes	NN	O	O
.	NN	O	O

The	NN	O	O
specificity	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
cell-free	NN	O	B-protein
synthesized	NN	O	I-protein
hGR	NN	O	I-protein
was	NN	O	O
analyzed	NN	O	O
further	NN	O	O
by	NN	O	O
examining	NN	O	O
the	NN	O	O
abilities	NN	O	O
of	NN	O	O
various	NN	O	B-DNA
DNAs	NN	O	I-DNA
to	NN	O	O
compete	NN	O	O
for	NN	O	O
binding	NN	O	O
to	NN	O	O
a	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
GRE	NN	O	B-DNA
found	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
mammary	NN	O	I-DNA
tumor	NN	O	I-DNA
virus-long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

Oligonucleotides	NN	O	O
containing	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
GRE	NN	O	I-DNA
were	NN	O	O
the	NN	O	O
most	NN	O	O
efficient	NN	O	O
competitors	NN	O	O
,	NN	O	O
and	NN	O	O
fragments	NN	O	O
containing	NN	O	O
regulatory	NN	O	B-DNA
sequences	NN	O	I-DNA
from	NN	O	O
glucocorticoid-repressible	NN	O	B-DNA
genes	NN	O	I-DNA
were	NN	O	O
somewhat	NN	O	O
competitive	NN	O	O
,	NN	O	O
whereas	NN	O	O
single	NN	O	O
stranded	NN	O	O
oligonucleotides	NN	O	O
were	NN	O	O
unable	NN	O	O
to	NN	O	O
compete	NN	O	O
for	NN	O	O
mouse	NN	O	B-DNA
mammary	NN	O	I-DNA
tumor	NN	O	I-DNA
virus-long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
except	NN	O	O
when	NN	O	O
competitor	NN	O	O
was	NN	O	O
present	NN	O	O
at	NN	O	O
extremely	NN	O	O
high	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Together	NN	O	O
these	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
hGR	NN	O	B-protein
synthesized	NN	O	O
in	NN	O	O
rabbit	NN	O	O
reticulocyte	NN	O	O
lysates	NN	O	O
displays	NN	O	O
many	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
properties	NN	O	O
,	NN	O	O
including	NN	O	O
GRE	NN	O	B-DNA
-specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
observed	NN	O	O
for	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
present	NN	O	O
in	NN	O	O
cytosolic	NN	O	O
extracts	NN	O	O
of	NN	O	O
mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
tissues	NN	O	O
.	NN	O	O

Similarities	NN	O	O
between	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
and	NN	O	O
RU486	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
antiglucocorticoid	NN	O	O
properties	NN	O	O
of	NN	O	O
RU486	NN	O	O
do	NN	O	O
not	NN	O	O
occur	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
specific	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptors	NN	O	I-protein
on	NN	O	O
alveolar	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
pulmonary	NN	O	O
granulomatous	NN	O	O
diseases	NN	O	O
.	NN	O	O

1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
produced	NN	O	O
at	NN	O	O
sites	NN	O	O
of	NN	O	O
granulomatous	NN	O	O
reactions	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
cell	NN	O	O
types	NN	O	O
that	NN	O	O
are	NN	O	O
targets	NN	O	O
for	NN	O	O
this	NN	O	O
immunoregulatory	NN	O	O
hormone	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
evaluated	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptors	NN	O	I-protein
on	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
and	NN	O	O
those	NN	O	O
recovered	NN	O	O
from	NN	O	O
the	NN	O	O
lung	NN	O	O
by	NN	O	O
bronchoalveolar	NN	O	O
lavage	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
pulmonary	NN	O	O
granulomatous	NN	O	O
diseases	NN	O	O
(	NN	O	O
tuberculosis	NN	O	O
and	NN	O	O
sarcoidosis	NN	O	O
)	NN	O	O
and	NN	O	O
from	NN	O	O
normal	NN	O	O
control	NN	O	O
subjects	NN	O	O
using	NN	O	O
combined	NN	O	O
autoradiographic	NN	O	O
and	NN	O	O
immunohistochemical	NN	O	O
techniques	NN	O	O
.	NN	O	O

Lavage	NN	O	O
T-lymphocytes	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
tuberculosis	NN	O	O
or	NN	O	O
with	NN	O	O
sarcoidosis	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
those	NN	O	O
from	NN	O	O
normal	NN	O	O
control	NN	O	O
subjects	NN	O	O
,	NN	O	O
expressed	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptors	NN	O	I-protein
as	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
excess	NN	O	O
unlabeled	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
unlabeled	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
D3	NN	O	O
(	NN	O	O
receptor-positive	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
sarcoidosis	NN	O	O
,	NN	O	O
20	NN	O	O
+/-	NN	O	O
12	NN	O	O
%	NN	O	O
;	NN	O	O
tuberculosis	NN	O	O
,	NN	O	O
31	NN	O	O
+/-	NN	O	O
17	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
blood	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
granulomatous	NN	O	O
diseases	NN	O	O
did	NN	O	O
not	NN	O	O
express	NN	O	O
detectable	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
percentage	NN	O	O
of	NN	O	O
lavage	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
expressing	NN	O	O
1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptors	NN	O	I-protein
was	NN	O	O
significantly	NN	O	O
greater	NN	O	O
for	NN	O	O
patients	NN	O	O
with	NN	O	O
tuberculosis	NN	O	O
presenting	NN	O	O
with	NN	O	O
isolated	NN	O	O
hilar	NN	O	O
adenopathy	NN	O	O
than	NN	O	O
for	NN	O	O
patients	NN	O	O
with	NN	O	O
pulmonary	NN	O	O
infiltrates	NN	O	O
and/or	NN	O	O
cavities	NN	O	O
.	NN	O	O

1	NN	O	B-protein
,	NN	O	I-protein
25	NN	O	I-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
2D3	NN	O	I-protein
receptors	NN	O	I-protein
were	NN	O	O
expressed	NN	O	O
to	NN	O	O
a	NN	O	O
greater	NN	O	O
extent	NN	O	O
on	NN	O	O
CD8+	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
than	NN	O	O
on	NN	O	O
CD4+	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
in	NN	O	O
sarcoidosis	NN	O	O
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
greater	NN	O	O
proportion	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_type
than	NN	O	O
of	NN	O	O
CD8+	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
tuberculosis	NN	O	O
were	NN	O	O
receptor-positive	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
support	NN	O	O
the	NN	O	O
conclusion	NN	O	O
that	NN	O	O
the	NN	O	O
interaction	NN	O	O
of	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
with	NN	O	O
its	NN	O	O
receptor	NN	O	O
on	NN	O	O
T-lymphocytes	NN	O	B-cell_type
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
granulomatous	NN	O	O
reactions	NN	O	O
,	NN	O	O
but	NN	O	O
because	NN	O	O
these	NN	O	O
receptors	NN	O	O
are	NN	O	O
expressed	NN	O	O
on	NN	O	O
different	NN	O	O
lymphocyte	NN	O	B-cell_type
populations	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
net	NN	O	O
effect	NN	O	O
of	NN	O	O
this	NN	O	O
potent	NN	O	O
immunoregulatory	NN	O	B-protein
molecule	NN	O	I-protein
is	NN	O	O
likely	NN	O	O
different	NN	O	O
in	NN	O	O
sarcoidosis	NN	O	O
and	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Role	NN	O	O
for	NN	O	O
the	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
2	NN	O	I-protein
in	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
switching	NN	O	O
during	NN	O	O
initial	NN	O	O
stages	NN	O	O
of	NN	O	O
infection	NN	O	O
.	NN	O	O

During	NN	O	O
latent	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
(	NN	O	O
EBV	NN	O	O
)	NN	O	O
infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
six	NN	O	B-protein
viral	NN	O	I-protein
nuclear	NN	O	I-protein
antigen	NN	O	I-protein
(	NN	O	O
EBNAs	NN	O	B-protein
)	NN	O	O
are	NN	O	O
expressed	NN	O	O
from	NN	O	O
long	NN	O	B-RNA
primary	NN	O	I-RNA
transcripts	NN	O	I-RNA
by	NN	O	O
means	NN	O	O
of	NN	O	O
alternative	NN	O	B-RNA
splicing	NN	O	I-RNA
and	NN	O	O
alternative	NN	O	B-RNA
polyadenylylation	NN	O	I-RNA
sites	NN	O	I-RNA
.	NN	O	O

These	NN	O	O
transcripts	NN	O	O
initiate	NN	O	O
from	NN	O	O
one	NN	O	O
of	NN	O	O
two	NN	O	O
promoters	NN	O	B-DNA
,	NN	O	O
Cp	NN	O	B-DNA
or	NN	O	O
Wp	NN	O	O
,	NN	O	O
that	NN	O	O
function	NN	O	O
in	NN	O	O
a	NN	O	O
mutually	NN	O	O
exclusive	NN	O	O
fashion	NN	O	O
.	NN	O	O

Wp	NN	O	O
is	NN	O	O
exclusively	NN	O	O
utilized	NN	O	O
during	NN	O	O
the	NN	O	O
initial	NN	O	O
stages	NN	O	O
of	NN	O	O
infection	NN	O	O
of	NN	O	O
primary	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
followed	NN	O	O
by	NN	O	O
a	NN	O	O
switch	NN	O	O
to	NN	O	O
Cp	NN	O	B-DNA
usage	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
have	NN	O	O
been	NN	O	O
extended	NN	O	O
to	NN	O	O
show	NN	O	O
that	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
a	NN	O	O
mutant	NN	O	O
EBV	NN	O	O
strain	NN	O	O
lacking	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
EBNA	NN	O	B-protein
2	NN	O	I-protein
fails	NN	O	O
to	NN	O	O
switch	NN	O	O
from	NN	O	O
Wp	NN	O	O
to	NN	O	O
Cp	NN	O	B-DNA
usage	NN	O	O
in	NN	O	O
primary	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
although	NN	O	O
the	NN	O	O
virus	NN	O	O
contains	NN	O	O
a	NN	O	O
functional	NN	O	O
Cp	NN	O	B-DNA
;	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
a	NN	O	O
region	NN	O	O
from	NN	O	O
-429	NN	O	B-DNA
to	NN	O	I-DNA
-245	NN	O	I-DNA
base	NN	O	I-DNA
pairs	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
Cp	NN	O	B-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
Cp	NN	O	B-DNA
activity	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
upstream	NN	O	B-DNA
and	NN	O	I-DNA
downstream	NN	O	I-DNA
sequences	NN	O	I-DNA
;	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
this	NN	O	O
region	NN	O	O
contains	NN	O	O
an	NN	O	O
EBNA	NN	O	B-DNA
2-dependent	NN	O	I-DNA
enhancer	NN	O	I-DNA
;	NN	O	O
and	NN	O	O
(	NN	O	O
iv	NN	O	O
)	NN	O	O
DNase	NN	O	B-protein
I	NN	O	I-protein
protection	NN	O	O
employing	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
revealed	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	B-DNA
footprint	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
EBNA	NN	O	B-DNA
2-dependent	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
support	NN	O	O
a	NN	O	O
model	NN	O	O
for	NN	O	O
viral	NN	O	B-DNA
promoter	NN	O	I-DNA
switching	NN	O	O
during	NN	O	O
the	NN	O	O
initial	NN	O	O
stages	NN	O	O
of	NN	O	O
infection	NN	O	O
in	NN	O	O
which	NN	O	O
Wp	NN	O	O
activity	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
EBNA	NN	O	B-protein
2	NN	O	I-protein
,	NN	O	O
followed	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
Cp	NN	O	B-DNA
through	NN	O	O
the	NN	O	O
EBNA	NN	O	B-DNA
2-dependent	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Immune	NN	O	O
response	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
HBx-antigen	NN	O	B-protein
of	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
virus	NN	O	O
.	NN	O	O

The	NN	O	O
hepatitis	NN	O	B-DNA
B	NN	O	I-DNA
virus	NN	O	I-DNA
genome	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
transactivator	NN	O	I-protein
protein	NN	O	I-protein
designated	NN	O	O
HBxAg	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
investigated	NN	O	O
whether	NN	O	O
this	NN	O	O
antigen	NN	O	O
is	NN	O	O
a	NN	O	O
target	NN	O	O
structure	NN	O	O
for	NN	O	O
human	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
.	NN	O	O

Using	NN	O	O
recombinant	NN	O	B-protein
HBxAg	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
we	NN	O	O
found	NN	O	O
HBxAg	NN	O	B-protein
-specific	NN	O	O
stimulation	NN	O	O
of	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
virus	NN	O	O
infection	NN	O	O
(	NN	O	O
6	NN	O	O
of	NN	O	O
6	NN	O	O
)	NN	O	O
and	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
B	NN	O	O
virus	NN	O	O
infection	NN	O	O
(	NN	O	O
6	NN	O	O
of	NN	O	O
17	NN	O	O
)	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
healthy	NN	O	O
individuals	NN	O	O
.	NN	O	O

With	NN	O	O
HBxAg	NN	O	B-protein
-specific	NN	O	O
synthetic	NN	O	O
polypeptides	NN	O	O
,	NN	O	O
several	NN	O	O
T-cell	NN	O	O
epitopes	NN	O	O
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Most	NN	O	O
were	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
carboxyterminal	NN	O	B-protein
half	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HBxAg	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Five	NN	O	O
T-cell	NN	O	B-cell_line
clones	NN	O	I-cell_line
specific	NN	O	O
for	NN	O	O
a	NN	O	O
T-cell	NN	O	B-protein
epitope	NN	O	I-protein
located	NN	O	O
at	NN	O	O
the	NN	O	O
carboxyterminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
HBxAg	NN	O	B-protein
were	NN	O	O
established	NN	O	O
and	NN	O	O
found	NN	O	O
to	NN	O	O
belong	NN	O	O
to	NN	O	O
the	NN	O	O
CD2/CD4-positive	NN	O	B-cell_line
,	NN	O	I-cell_line
CD8-negative	NN	O	I-cell_line
subtype	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
data	NN	O	O
establish	NN	O	O
for	NN	O	O
the	NN	O	O
first	NN	O	O
time	NN	O	O
HBxAg	NN	O	B-protein
as	NN	O	O
an	NN	O	O
antigen	NN	O	O
in	NN	O	O
the	NN	O	O
cellular	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Changes	NN	O	O
in	NN	O	O
leucocytic	NN	O	B-protein
estrogen	NN	O	I-protein
receptor	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
gynecomastia	NN	O	O
]	NN	O	O

The	NN	O	O
number	NN	O	O
of	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
in	NN	O	O
13	NN	O	O
men	NN	O	O
with	NN	O	O
gynecomastia	NN	O	O
were	NN	O	O
measured	NN	O	O
by	NN	O	O
radioligand	NN	O	O
binding	NN	O	O
method.	NN	O	O
The	NN	O	O
results	NN	O	O
were	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
13	NN	O	O
sex-and	NN	O	O
age-matched	NN	O	O
healthy	NN	O	O
subjects	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
in	NN	O	O
leucocytes	NN	O	B-cell_type
was	NN	O	O
significantly	NN	O	O
increased	NN	O	O
in	NN	O	O
gynecomastia	NN	O	O
(	NN	O	O
Rs	NN	O	O
of	NN	O	O
leucocytes	NN	O	B-cell_type
were	NN	O	O
1054	NN	O	O
+/-	NN	O	O
254	NN	O	O
sites/cell	NN	O	O
)	NN	O	O
.	NN	O	O

It	NN	O	O
suggested	NN	O	O
that	NN	O	O
increase	NN	O	O
of	NN	O	O
ER	NN	O	O
levels	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
gynecomastia	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
independent	NN	O	I-DNA
cis-acting	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
responsive	NN	O	O
to	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
rate	NN	O	O
of	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
directed	NN	O	O
by	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	O
)	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
increases	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
mitogenic	NN	O	O
stimuli	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
may	NN	O	O
be	NN	O	O
governed	NN	O	O
by	NN	O	O
two	NN	O	O
independent	NN	O	O
sequences	NN	O	O
located	NN	O	O
5	NN	O	O
'	NN	O	O
to	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
transcription	NN	O	O
initiation	NN	O	O
sequences	NN	O	O
that	NN	O	O
bind	NN	O	O
either	NN	O	O
NFAT-1	NN	O	B-protein
or	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
rate	NN	O	O
of	NN	O	O
LTR	NN	O	B-DNA
-directed	NN	O	O
gene	NN	O	O
expression	NN	O	O
increased	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
treatment	NN	O	O
with	NN	O	O
either	NN	O	O
a	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
or	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
if	NN	O	O
either	NN	O	O
the	NN	O	O
NFAT-1	NN	O	B-protein
or	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
were	NN	O	O
deleted	NN	O	O
,	NN	O	O
but	NN	O	O
failed	NN	O	O
to	NN	O	O
respond	NN	O	O
to	NN	O	O
these	NN	O	O
mitogenic	NN	O	O
stimuli	NN	O	O
if	NN	O	O
both	NN	O	O
sequences	NN	O	O
were	NN	O	O
absent	NN	O	O
.	NN	O	O

The	NN	O	O
HIV-1	NN	O	O
mutant	NN	O	O
virus	NN	O	O
containing	NN	O	O
both	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
NFAT-1	NN	O	I-DNA
deletion	NN	O	I-DNA
was	NN	O	O
able	NN	O	O
to	NN	O	O
replicate	NN	O	O
although	NN	O	O
at	NN	O	O
a	NN	O	O
much	NN	O	O
decreased	NN	O	O
growth	NN	O	O
rate	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
deletion	NN	O	O
of	NN	O	O
NFAT-1	NN	O	B-protein
alone	NN	O	O
increased	NN	O	O
the	NN	O	O
viral	NN	O	O
growth	NN	O	O
rate	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Neither	NN	O	O
deletion	NN	O	O
of	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
nor	NN	O	O
deletion	NN	O	O
of	NN	O	O
NFAT-1	NN	O	B-protein
decreased	NN	O	O
activation	NN	O	O
of	NN	O	O
viral	NN	O	O
replication	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
.	NN	O	O

-DOCSTART-	O

cAMP-dependent	NN	O	O
regulation	NN	O	O
of	NN	O	O
proenkephalin	NN	O	O
by	NN	O	O
JunD	NN	O	B-protein
and	NN	O	O
JunB	NN	O	B-protein
:	NN	O	O
positive	NN	O	O
and	NN	O	O
negative	NN	O	O
effects	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
JunD	NN	O	B-protein
,	NN	O	O
a	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
activates	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
proenkephalin	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
fashion	NN	O	O
that	NN	O	O
is	NN	O	O
completely	NN	O	O
dependent	NN	O	O
upon	NN	O	O
the	NN	O	O
cAMP-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
proenkephalin	NN	O	O
transcription	NN	O	O
by	NN	O	O
JunD	NN	O	B-protein
is	NN	O	O
dependent	NN	O	O
upon	NN	O	O
a	NN	O	O
previously	NN	O	O
characterized	NN	O	O
cAMP-	NN	O	B-DNA
,	NN	O	I-DNA
phorbol	NN	O	I-DNA
ester-	NN	O	I-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
Ca	NN	O	I-DNA
(	NN	O	I-DNA
2+	NN	O	I-DNA
)	NN	O	I-DNA
-inducible	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
JunD	NN	O	B-protein
is	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
as	NN	O	O
a	NN	O	O
homodimer	NN	O	B-protein
.	NN	O	O

Another	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
transcription	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
JunB	NN	O	B-protein
,	NN	O	O
is	NN	O	O
shown	NN	O	O
to	NN	O	O
inhibit	NN	O	O
activation	NN	O	O
mediated	NN	O	O
by	NN	O	O
JunD	NN	O	B-protein
.	NN	O	O

As	NN	O	O
a	NN	O	O
homodimer	NN	O	O
JunB	NN	O	B-protein
is	NN	O	O
unable	NN	O	O
to	NN	O	O
bind	NN	O	O
the	NN	O	O
enhancer	NN	O	O
;	NN	O	O
however	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
c-Fos	NN	O	B-protein
,	NN	O	O
high-affinity	NN	O	O
binding	NN	O	O
is	NN	O	O
observed	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
JunD	NN	O	B-protein
is	NN	O	O
shown	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	O
linked	NN	O	O
to	NN	O	O
both	NN	O	O
cAMP	NN	O	O
and	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
response	NN	O	O
elements	NN	O	O
in	NN	O	O
a	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
-dependent	NN	O	O
fashion	NN	O	O
,	NN	O	O
further	NN	O	O
blurring	NN	O	O
the	NN	O	O
distinction	NN	O	O
between	NN	O	O
these	NN	O	O
response	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
an	NN	O	O
AP-1-related	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
cAMP-dependent	NN	O	O
second-messenger	NN	O	O
pathway	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
JunD	NN	O	B-protein
and	NN	O	O
other	NN	O	O
AP-1-related	NN	O	B-protein
proteins	NN	O	I-protein
may	NN	O	O
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
cAMP-dependent	NN	O	O
intracellular	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Plasma	NN	O	O
cortisol	NN	O	O
concentration	NN	O	O
and	NN	O	O
blood	NN	O	O
leukocyte	NN	O	O
content	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
patients	NN	O	O
with	NN	O	O
deficiency-cold	NN	O	O
vs	NN	O	O
deficiency-heat	NN	O	O
syndromes	NN	O	O
]	NN	O	O

Plasma	NN	O	O
cortisol	NN	O	O
concentration	NN	O	O
and	NN	O	O
blood	NN	O	O
leukocyte	NN	O	O
content	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GCR	NN	O	B-protein
)	NN	O	O
were	NN	O	O
assayed	NN	O	O
in	NN	O	O
20	NN	O	O
patients	NN	O	O
with	NN	O	O
deficiency	NN	O	O
syndromes	NN	O	O
,	NN	O	O
10	NN	O	O
cold	NN	O	O
in	NN	O	O
property	NN	O	O
(	NN	O	O
deficiency-cold	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
other	NN	O	O
10	NN	O	O
hot	NN	O	O
in	NN	O	O
property	NN	O	O
(	NN	O	O
deficiency-heat	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
also	NN	O	O
in	NN	O	O
10	NN	O	O
healthy	NN	O	O
individuals	NN	O	O
as	NN	O	O
normal	NN	O	O
control	NN	O	O
for	NN	O	O
the	NN	O	O
purpose	NN	O	O
of	NN	O	O
investigating	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
cold	NN	O	O
and	NN	O	O
heat	NN	O	O
syndromes	NN	O	O
.	NN	O	O

As	NN	O	O
a	NN	O	O
result	NN	O	O
,	NN	O	O
the	NN	O	O
cases	NN	O	O
of	NN	O	O
deficiency-cold	NN	O	O
syndrome	NN	O	O
(	NN	O	O
DCS	NN	O	O
)	NN	O	O
had	NN	O	O
a	NN	O	O
normal	NN	O	O
concentration	NN	O	O
of	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
but	NN	O	O
a	NN	O	O
lowered	NN	O	O
content	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
in	NN	O	O
leukocytes	NN	O	B-cell_type
when	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
normal	NN	O	O
control	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.05	NN	O	O
)	NN	O	O
;	NN	O	O
the	NN	O	O
cases	NN	O	O
of	NN	O	O
deficiency-heat	NN	O	O
syndrome	NN	O	O
(	NN	O	O
DHS	NN	O	O
)	NN	O	O
had	NN	O	O
a	NN	O	O
higher	NN	O	O
concentration	NN	O	O
of	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
than	NN	O	O
the	NN	O	O
normal	NN	O	O
control	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.05	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
slightly	NN	O	O
higher	NN	O	O
content	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
in	NN	O	O
leukocytes	NN	O	B-cell_type
.	NN	O	O

It	NN	O	O
was	NN	O	O
concluded	NN	O	O
that	NN	O	O
the	NN	O	O
DCS	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
diminished	NN	O	O
biological	NN	O	O
effects	NN	O	O
of	NN	O	O
adrenocortical	NN	O	O
activity	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
DHS	NN	O	O
,	NN	O	O
by	NN	O	O
augmented	NN	O	O
biological	NN	O	O
effects	NN	O	O
of	NN	O	O
adrenocortical	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
role	NN	O	O
of	NN	O	O
jun	NN	O	B-DNA
and	NN	O	I-DNA
fos	NN	O	I-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
members	NN	O	I-DNA
in	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
induced	NN	O	O
hemopoietic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Terminal	NN	O	O
differentiation	NN	O	O
of	NN	O	O
the	NN	O	O
leukemic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
U-937	NN	O	B-cell_line
and	NN	O	O
HL-60	NN	O	B-cell_line
by	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
marked	NN	O	O
changes	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
jun	NN	O	B-DNA
and	NN	O	I-DNA
fos	NN	O	I-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
members	NN	O	I-DNA
is	NN	O	O
induced	NN	O	O
with	NN	O	O
variable	NN	O	O
kinetics	NN	O	O
during	NN	O	O
12-O-tetradecanoylphorbol-13-acetate	NN	O	O
induced	NN	O	O
differentiation	NN	O	O
,	NN	O	O
with	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
best	NN	O	O
paralleling	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
generation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
complexes	NN	O	I-protein
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
closely	NN	O	O
parallels	NN	O	O
morphological	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
these	NN	O	O
complexes	NN	O	O
to	NN	O	O
regulate	NN	O	O
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
increased	NN	O	O
transcription	NN	O	O
from	NN	O	O
an	NN	O	O
AP-1	NN	O	B-DNA
driven	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
and	NN	O	O
marked	NN	O	O
increases	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
endogenous	NN	O	B-DNA
AP-1	NN	O	I-DNA
regulated	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

Differentiation	NN	O	O
assays	NN	O	O
using	NN	O	O
water	NN	O	O
soluble	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
reveal	NN	O	O
that	NN	O	O
differentiation	NN	O	O
becomes	NN	O	O
irreversible	NN	O	O
soon	NN	O	O
after	NN	O	O
AP-1	NN	O	B-protein
appears	NN	O	O
.	NN	O	O

This	NN	O	O
tight	NN	O	O
correlation	NN	O	O
between	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
,	NN	O	O
the	NN	O	O
generation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
and	NN	O	O
differentiation	NN	O	O
suggests	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
for	NN	O	O
this	NN	O	O
gene	NN	O	O
and	NN	O	O
transcriptional	NN	O	B-protein
complex	NN	O	I-protein
during	NN	O	O
this	NN	O	O
process	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	O
association	NN	O	O
of	NN	O	O
a	NN	O	O
T-cell	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
blocked	NN	O	O
by	NN	O	O
FK-506	NN	O	O
and	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
[	NN	O	O
see	NN	O	O
comments	NN	O	O
]	NN	O	O

Cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
inhibit	NN	O	O
T-	NN	O	O
and	NN	O	O
B-cell	NN	O	O
activation	NN	O	O
and	NN	O	O
other	NN	O	O
processes	NN	O	O
essential	NN	O	O
to	NN	O	O
an	NN	O	O
effective	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
these	NN	O	O
drugs	NN	O	O
disrupt	NN	O	O
an	NN	O	O
unknown	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
transmission	NN	O	O
of	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
antigen	NN	O	I-protein
receptor	NN	O	I-protein
to	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
that	NN	O	O
coordinate	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

The	NN	O	O
putative	NN	O	B-protein
intracellular	NN	O	I-protein
receptors	NN	O	I-protein
for	NN	O	O
FK506	NN	O	O
and	NN	O	O
cyclosporin	NN	O	O
are	NN	O	O
cis-trans	NN	O	O
prolyl	NN	O	O
isomerases	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
the	NN	O	O
drug	NN	O	O
inhibits	NN	O	O
isomerase	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
but	NN	O	O
studies	NN	O	O
with	NN	O	O
other	NN	O	O
prolyl	NN	O	O
isomerase	NN	O	B-protein
inhibitors	NN	O	O
and	NN	O	O
analysis	NN	O	O
of	NN	O	O
cyclosporin-resistant	NN	O	B-cell_line
mutants	NN	O	I-cell_line
in	NN	O	O
yeast	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
the	NN	O	O
drug	NN	O	O
result	NN	O	O
from	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
an	NN	O	O
inhibitory	NN	O	B-protein
complex	NN	O	I-protein
between	NN	O	O
the	NN	O	O
drug	NN	O	O
and	NN	O	O
isomerase	NN	O	B-protein
,	NN	O	O
and	NN	O	O
not	NN	O	O
from	NN	O	O
inhibition	NN	O	O
of	NN	O	O
isomerase	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

A	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
early	NN	O	O
T-cell	NN	O	O
gene	NN	O	O
activation	NN	O	O
,	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
specific	NN	O	O
target	NN	O	O
of	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
action	NN	O	O
because	NN	O	O
transcription	NN	O	O
directed	NN	O	O
by	NN	O	O
this	NN	O	O
protein	NN	O	O
is	NN	O	O
blocked	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
treated	NN	O	O
with	NN	O	O
these	NN	O	O
drugs	NN	O	O
,	NN	O	O
with	NN	O	O
little	NN	O	O
or	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
NF-AT	NN	O	B-protein
is	NN	O	O
formed	NN	O	O
when	NN	O	O
a	NN	O	O
signal	NN	O	O
from	NN	O	O
the	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
induces	NN	O	O
a	NN	O	O
pre-existing	NN	O	O
cytoplasmic	NN	O	O
subunit	NN	O	O
to	NN	O	O
translocate	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
and	NN	O	O
combine	NN	O	O
with	NN	O	O
a	NN	O	O
newly	NN	O	O
synthesized	NN	O	O
nuclear	NN	O	O
subunit	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
.	NN	O	O

FK506	NN	O	O
and	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
block	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
component	NN	O	I-protein
without	NN	O	O
affecting	NN	O	O
synthesis	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
subunit	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
in	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
.	NN	O	O

OBJECTIVE	NN	O	O
:	NN	O	O
The	NN	O	O
aim	NN	O	O
was	NN	O	O
to	NN	O	O
explore	NN	O	O
the	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
during	NN	O	O
hypercortisolaemia	NN	O	O
in	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
.	NN	O	O

DESIGN	NN	O	O
:	NN	O	O
Urine	NN	O	O
and	NN	O	O
plasma	NN	O	O
samples	NN	O	O
were	NN	O	O
obtained	NN	O	O
for	NN	O	O
cortisol	NN	O	O
determination	NN	O	O
and	NN	O	O
blood	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
isolated	NN	O	O
for	NN	O	O
receptor	NN	O	O
binding	NN	O	O
studies	NN	O	O
.	NN	O	O

PATIENTS	NN	O	O
:	NN	O	O
Sixteen	NN	O	O
anorexic	NN	O	O
patients	NN	O	O
,	NN	O	O
aged	NN	O	O
16-27	NN	O	O
years	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
body	NN	O	O
mass	NN	O	O
index	NN	O	O
of	NN	O	O
14.2	NN	O	O
+/-	NN	O	O
2.0	NN	O	O
(	NN	O	O
ranging	NN	O	O
from	NN	O	O
11.1	NN	O	O
to	NN	O	O
17.4	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
15	NN	O	O
normal	NN	O	O
women	NN	O	O
were	NN	O	O
studied	NN	O	O
.	NN	O	O

Six	NN	O	O
patients	NN	O	O
were	NN	O	O
reinvestigated	NN	O	O
after	NN	O	O
a	NN	O	O
significant	NN	O	O
weight	NN	O	O
gain	NN	O	O
.	NN	O	O

MEASUREMENTS	NN	O	O
:	NN	O	O
The	NN	O	O
binding	NN	O	O
capacity	NN	O	O
and	NN	O	O
affinity	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
were	NN	O	O
measured	NN	O	O
with	NN	O	O
dexamethasone	NN	O	O
as	NN	O	O
ligand	NN	O	O
on	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
In	NN	O	O
patients	NN	O	O
,	NN	O	O
both	NN	O	O
total	NN	O	O
and	NN	O	O
free	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
were	NN	O	O
higher	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
normal	NN	O	O
women	NN	O	O
,	NN	O	O
as	NN	O	O
was	NN	O	O
their	NN	O	O
urinary	NN	O	O
free	NN	O	O
cortisol	NN	O	O
;	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
per	NN	O	O
cell	NN	O	O
(	NN	O	O
Ro	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
binding	NN	O	O
affinity	NN	O	O
(	NN	O	O
Kd	NN	O	O
)	NN	O	O
for	NN	O	O
dexamethasone	NN	O	O
were	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
not	NN	O	O
significantly	NN	O	O
different	NN	O	O
(	NN	O	O
Ro	NN	O	O
:	NN	O	O
7687	NN	O	O
+/-	NN	O	O
1750	NN	O	O
vs	NN	O	O
7347	NN	O	O
+/-	NN	O	O
1285	NN	O	O
sites/cell	NN	O	O
;	NN	O	O
Kd	NN	O	O
:	NN	O	O
7.7	NN	O	O
+/-	NN	O	O
2.4	NN	O	O
vs	NN	O	O
7.4	NN	O	O
+/-	NN	O	O
1.7	NN	O	O
nM	NN	O	O
at	NN	O	O
24	NN	O	O
degrees	NN	O	O
C	NN	O	O
)	NN	O	O
.	NN	O	O

After	NN	O	O
weight	NN	O	O
gain	NN	O	O
(	NN	O	O
14	NN	O	O
+/-	NN	O	O
2	NN	O	O
to	NN	O	O
16	NN	O	O
+/-	NN	O	O
2	NN	O	O
kg/m2	NN	O	O
)	NN	O	O
,	NN	O	O
receptor	NN	O	O
numbers	NN	O	O
were	NN	O	O
8421	NN	O	O
+/-	NN	O	O
2126	NN	O	O
(	NN	O	O
pre	NN	O	O
)	NN	O	O
and	NN	O	O
9011	NN	O	O
+/-	NN	O	O
500	NN	O	O
(	NN	O	O
post	NN	O	O
)	NN	O	O
sites/cell	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
not	NN	O	O
significantly	NN	O	O
different	NN	O	O
(	NN	O	O
P	NN	O	O
greater	NN	O	O
than	NN	O	O
0.2	NN	O	O
)	NN	O	O
;	NN	O	O
the	NN	O	O
Kd	NN	O	O
was	NN	O	O
unchanged	NN	O	O
(	NN	O	O
9.3	NN	O	O
+/-	NN	O	O
2.6	NN	O	O
vs	NN	O	O
9.2	NN	O	O
+/-	NN	O	O
2.4	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
Hypercortisolaemia	NN	O	O
does	NN	O	O
not	NN	O	O
down-regulate	NN	O	O
the	NN	O	O
lymphocyte	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
anorexia	NN	O	O
nervosa	NN	O	O
and	NN	O	O
a	NN	O	O
post-receptor	NN	O	O
defect	NN	O	O
might	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
peripheral	NN	O	O
tissue	NN	O	O
resistance	NN	O	O
to	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
hormones	NN	O	O
in	NN	O	O
undernutrition	NN	O	O
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
phosphatases	NN	O	I-protein
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
induces	NN	O	O
AP1	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

To	NN	O	O
examine	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
phosphatases	NN	O	I-protein
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
,	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
treated	NN	O	O
with	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
,	NN	O	O
an	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
type	NN	O	B-protein
1	NN	O	I-protein
and	NN	O	I-protein
2A	NN	O	I-protein
phosphatases	NN	O	I-protein
,	NN	O	O
and	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
AP1	NN	O	B-protein
as	NN	O	O
a	NN	O	O
measure	NN	O	O
of	NN	O	O
early	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Okadaic	NN	O	O
acid	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
potent	NN	O	O
inducer	NN	O	O
of	NN	O	O
AP1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
such	NN	O	O
as	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
(	NN	O	O
PMA	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
AP1	NN	O	B-protein
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
occurs	NN	O	O
predominantly	NN	O	O
by	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
jun	NN	O	B-DNA
and	NN	O	I-DNA
fos	NN	O	I-DNA
family	NN	O	I-DNA
of	NN	O	O
proto-oncogenes	NN	O	B-DNA
.	NN	O	O

Surprisingly	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
phytohemagglutinin	NN	O	B-protein
further	NN	O	O
enhanced	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
AP1	NN	O	B-protein
,	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
PMA	NN	O	O
inhibited	NN	O	O
it	NN	O	O
.	NN	O	O

Okadaic	NN	O	O
acid	NN	O	O
treatment	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
dramatically	NN	O	O
increase	NN	O	O
mRNA	NN	O	B-RNA
transcripts	NN	O	I-RNA
of	NN	O	O
the	NN	O	O
jun	NN	O	B-DNA
family	NN	O	I-DNA
of	NN	O	I-DNA
proto-oncogenes	NN	O	I-DNA
including	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	O
junD	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
junB	NN	O	B-DNA
and	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
the	NN	O	O
fos	NN	O	B-DNA
family	NN	O	I-DNA
including	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
fra-1	NN	O	B-DNA
.	NN	O	O

By	NN	O	O
comparison	NN	O	O
,	NN	O	O
PMA	NN	O	O
is	NN	O	O
a	NN	O	O
very	NN	O	O
inefficient	NN	O	O
inducer	NN	O	O
of	NN	O	O
the	NN	O	O
jun	NN	O	B-DNA
gene	NN	O	I-DNA
family	NN	O	I-DNA
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
its	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
AP1	NN	O	B-protein
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
,	NN	O	O
PMA	NN	O	O
inhibits	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
.	NN	O	O

Transfection	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
confirmed	NN	O	O
the	NN	O	O
marked	NN	O	O
difference	NN	O	O
between	NN	O	O
PMA	NN	O	O
and	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
in	NN	O	O
inducing	NN	O	O
c-jun	NN	O	B-DNA
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
AP1	NN	O	B-protein
by	NN	O	O
okadaic	NN	O	O
acid	NN	O	O
suggests	NN	O	O
that	NN	O	O
protein	NN	O	B-protein
phosphatases	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	I-protein
2A	NN	O	I-protein
(	NN	O	O
PP1	NN	O	B-protein
and	NN	O	O
PP2A	NN	O	B-protein
)	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
as	NN	O	O
important	NN	O	O
negative	NN	O	O
regulators	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
AP1	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
of	NN	O	O
murine	NN	O	B-protein
TCF-1	NN	O	I-protein
,	NN	O	O
a	NN	O	O
T	NN	O	O
cell-specific	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
interacting	NN	O	O
with	NN	O	O
functional	NN	O	B-DNA
motifs	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
CD3-epsilon	NN	O	B-DNA
and	NN	O	I-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
enhancers	NN	O	I-DNA
.	NN	O	O

CD3	NN	O	B-protein
-epsilon	NN	O	O
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
confined	NN	O	O
to	NN	O	O
the	NN	O	O
T	NN	O	B-cell_line
cell	NN	O	I-cell_line
lineage	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
identified	NN	O	O
and	NN	O	O
cloned	NN	O	O
a	NN	O	O
human	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
TCF-1	NN	O	B-protein
,	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
functional	NN	O	O
element	NN	O	O
in	NN	O	O
the	NN	O	O
T	NN	O	O
lymphocyte-specific	NN	O	O
enhancer	NN	O	O
of	NN	O	O
CD3-epsilon	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
human	NN	O	O
cell	NN	O	O
lines	NN	O	O
,	NN	O	O
TCF-1	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
restricted	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
lineage	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

TCF-1	NN	O	B-protein
belonged	NN	O	O
to	NN	O	O
a	NN	O	O
novel	NN	O	O
family	NN	O	O
of	NN	O	O
genes	NN	O	O
that	NN	O	O
contain	NN	O	O
the	NN	O	O
so-called	NN	O	O
high	NN	O	O
mobility	NN	O	B-DNA
group	NN	O	I-DNA
1	NN	O	I-DNA
(	NN	O	I-DNA
HMG	NN	O	I-DNA
)	NN	O	I-DNA
box	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
cloning	NN	O	O
of	NN	O	O
murine	NN	O	B-protein
TCF-1	NN	O	I-protein
.	NN	O	O

Two	NN	O	O
splice	NN	O	O
alternatives	NN	O	O
were	NN	O	O
identified	NN	O	O
that	NN	O	O
were	NN	O	O
not	NN	O	O
previously	NN	O	O
observed	NN	O	O
in	NN	O	O
human	NN	O	B-protein
TCF-1	NN	O	I-protein
.	NN	O	O

Murine	NN	O	B-protein
and	NN	O	I-protein
human	NN	O	I-protein
TCF-1	NN	O	I-protein
displayed	NN	O	O
a	NN	O	O
95.5	NN	O	O
%	NN	O	O
overall	NN	O	O
amino	NN	O	O
acid	NN	O	O
homology	NN	O	O
.	NN	O	O

Recombinant	NN	O	B-protein
murine	NN	O	I-protein
and	NN	O	I-protein
human	NN	O	I-protein
TCF-1	NN	O	I-protein
recognized	NN	O	O
the	NN	O	O
same	NN	O	O
sequence	NN	O	O
motif	NN	O	O
in	NN	O	O
the	NN	O	O
CD3-epsilon	NN	O	B-DNA
enhancer	NN	O	I-DNA
as	NN	O	O
judged	NN	O	O
by	NN	O	O
gel	NN	O	O
retardation	NN	O	O
and	NN	O	O
methylation	NN	O	O
interference	NN	O	O
assays	NN	O	O
.	NN	O	O

With	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
cDNA	NN	O	I-DNA
clones	NN	O	I-DNA
several	NN	O	O
aspects	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
were	NN	O	O
analyzed	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
deletion	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
a	NN	O	O
region	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
containing	NN	O	O
the	NN	O	O
HMG	NN	O	B-DNA
box	NN	O	I-DNA
was	NN	O	O
sufficient	NN	O	O
for	NN	O	O
sequence-specific	NN	O	O
binding	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
by	NN	O	O
high	NN	O	O
stringency	NN	O	O
Northern	NN	O	O
blotting	NN	O	O
and	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
,	NN	O	O
TCF-1	NN	O	B-protein
expression	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
confined	NN	O	O
to	NN	O	O
the	NN	O	O
thymus	NN	O	O
and	NN	O	O
to	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
areas	NN	O	O
of	NN	O	O
the	NN	O	O
spleen	NN	O	O
.	NN	O	O

Third	NN	O	O
,	NN	O	O
TCF-1	NN	O	B-protein
bound	NN	O	O
specifically	NN	O	O
to	NN	O	O
a	NN	O	O
functional	NN	O	O
T	NN	O	B-DNA
cell-specific	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
T	NN	O	B-DNA
cell	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
TCR-alpha	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
T	NN	O	O
lineage-specific	NN	O	O
expression	NN	O	O
and	NN	O	O
the	NN	O	O
affinity	NN	O	O
for	NN	O	O
functional	NN	O	O
motifs	NN	O	O
in	NN	O	O
the	NN	O	O
TCR-alpha	NN	O	B-DNA
and	NN	O	I-DNA
CD3-epsilon	NN	O	I-DNA
enhancers	NN	O	I-DNA
imply	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
for	NN	O	O
TCF-1	NN	O	B-protein
in	NN	O	O
the	NN	O	O
establishment	NN	O	O
of	NN	O	O
the	NN	O	O
mature	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
phenotype	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
homeobox	NN	O	I-DNA
gene	NN	O	I-DNA
that	NN	O	O
resembles	NN	O	O
a	NN	O	O
diverged	NN	O	B-DNA
Drosophila	NN	O	I-DNA
homeobox	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
activated	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

A	NN	O	O
new	NN	O	O
homeobox	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
HB24	NN	O	B-DNA
,	NN	O	O
has	NN	O	O
been	NN	O	O
isolated	NN	O	O
from	NN	O	O
a	NN	O	O
human	NN	O	O
B-lymphocyte	NN	O	O
cDNA	NN	O	O
library	NN	O	O
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
polyadenylated	NN	O	B-RNA
RNA	NN	O	I-RNA
purified	NN	O	O
from	NN	O	O
activated	NN	O	B-cell_type
human	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
revealed	NN	O	O
a	NN	O	O
single	NN	O	B-RNA
mRNA	NN	O	I-RNA
transcript	NN	O	I-RNA
of	NN	O	O
approximately	NN	O	O
2.3	NN	O	O
kb	NN	O	O
.	NN	O	O

Two	NN	O	O
cDNA	NN	O	B-DNA
clones	NN	O	I-DNA
were	NN	O	O
sequenced	NN	O	O
and	NN	O	O
provided	NN	O	O
2	NN	O	O
,	NN	O	O
250	NN	O	O
nucleotides	NN	O	O
(	NN	O	O
nt	NN	O	O
)	NN	O	O
of	NN	O	O
DNA	NN	O	B-DNA
sequence	NN	O	I-DNA
information	NN	O	O
.	NN	O	O

There	NN	O	O
is	NN	O	O
a	NN	O	O
single	NN	O	O
methionine	NN	O	B-DNA
codon-initiated	NN	O	I-DNA
open	NN	O	I-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
of	NN	O	O
1	NN	O	O
,	NN	O	O
458	NN	O	O
nt	NN	O	O
in	NN	O	O
frame	NN	O	O
with	NN	O	O
a	NN	O	O
homeobox	NN	O	B-DNA
and	NN	O	O
a	NN	O	O
CAX	NN	O	B-DNA
repeat	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
the	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
is	NN	O	O
predicted	NN	O	O
to	NN	O	O
encode	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
51	NN	O	O
,	NN	O	O
659	NN	O	O
daltons	NN	O	O
.	NN	O	O

When	NN	O	O
the	NN	O	O
homeodomain	NN	O	B-DNA
from	NN	O	O
HB24	NN	O	B-DNA
was	NN	O	O
compared	NN	O	O
to	NN	O	O
known	NN	O	O
mammalian	NN	O	B-DNA
and	NN	O	I-DNA
Drosophila	NN	O	I-DNA
homeodomains	NN	O	I-DNA
it	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
only	NN	O	O
moderately	NN	O	O
conserved	NN	O	O
,	NN	O	O
but	NN	O	O
when	NN	O	O
it	NN	O	O
was	NN	O	O
compared	NN	O	O
to	NN	O	O
a	NN	O	O
highly	NN	O	O
diverged	NN	O	O
Drosophila	NN	O	B-DNA
homeodomain	NN	O	I-DNA
,	NN	O	O
H2.0	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
80	NN	O	O
%	NN	O	O
identical	NN	O	O
.	NN	O	O

The	NN	O	O
HB24	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
absent	NN	O	O
or	NN	O	O
present	NN	O	O
at	NN	O	O
low	NN	O	O
levels	NN	O	O
in	NN	O	O
normal	NN	O	O
B	NN	O	O
and	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
;	NN	O	O
however	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
appropriate	NN	O	O
activation	NN	O	O
signal	NN	O	O
HB24	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
induced	NN	O	O
within	NN	O	O
several	NN	O	O
hours	NN	O	O
even	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cycloheximide	NN	O	O
.	NN	O	O

Characterization	NN	O	O
of	NN	O	O
HB24	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
lymphoid	NN	O	O
and	NN	O	O
select	NN	O	O
developing	NN	O	O
tissues	NN	O	O
was	NN	O	O
performed	NN	O	O
by	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
.	NN	O	O

Positive	NN	O	O
hybridization	NN	O	O
was	NN	O	O
found	NN	O	O
in	NN	O	O
thymus	NN	O	O
,	NN	O	O
tonsil	NN	O	O
,	NN	O	O
bone	NN	O	O
marrow	NN	O	O
,	NN	O	O
developing	NN	O	O
vessels	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
fetal	NN	O	O
brain	NN	O	O
.	NN	O	O

HB24	NN	O	B-DNA
is	NN	O	O
likely	NN	O	O
to	NN	O	O
have	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
certain	NN	O	O
developing	NN	O	O
tissues	NN	O	O
.	NN	O	O

-DOCSTART-	O

Platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
induces	NN	O	O
phospholipid	NN	O	O
turnover	NN	O	O
,	NN	O	O
calcium	NN	O	O
flux	NN	O	O
,	NN	O	O
arachidonic	NN	O	O
acid	NN	O	O
liberation	NN	O	O
,	NN	O	O
eicosanoid	NN	O	O
generation	NN	O	O
,	NN	O	O
and	NN	O	O
oncogene	NN	O	B-DNA
expression	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
mediator	NN	O	O
of	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
response	NN	O	O
.	NN	O	O

Studies	NN	O	O
of	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
have	NN	O	O
centered	NN	O	O
mainly	NN	O	O
around	NN	O	O
neutrophils	NN	O	B-cell_type
,	NN	O	O
monocytes	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
platelets	NN	O	B-cell_type
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
begin	NN	O	O
to	NN	O	O
uncover	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
on	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Employing	NN	O	O
the	NN	O	O
EBV-transformed	NN	O	B-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
SKW6.4	NN	O	B-cell_line
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
significantly	NN	O	O
alters	NN	O	O
membrane	NN	O	O
phospholipid	NN	O	O
metabolism	NN	O	O
indicated	NN	O	O
by	NN	O	O
the	NN	O	O
incorporation	NN	O	O
of	NN	O	O
32P	NN	O	O
into	NN	O	O
phosphatidylcholine	NN	O	O
,	NN	O	O
phosphatidylinositol	NN	O	O
,	NN	O	O
and	NN	O	O
phosphatidic	NN	O	O
acid	NN	O	O
but	NN	O	O
not	NN	O	O
significantly	NN	O	O
into	NN	O	O
phosphatidylethanolamine	NN	O	O
at	NN	O	O
concentrations	NN	O	O
ranging	NN	O	O
from	NN	O	O
10	NN	O	O
(	NN	O	O
-9	NN	O	O
)	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
M	NN	O	O
.	NN	O	O

The	NN	O	O
inactive	NN	O	O
precursor	NN	O	O
,	NN	O	O
lyso-platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
at	NN	O	O
a	NN	O	O
concentration	NN	O	O
as	NN	O	O
high	NN	O	O
as	NN	O	O
10	NN	O	O
(	NN	O	O
-7	NN	O	O
)	NN	O	O
M	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
membrane	NN	O	O
phospholipids	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
from	NN	O	O
10	NN	O	O
(	NN	O	O
-12	NN	O	O
)	NN	O	O
to	NN	O	O
10	NN	O	O
(	NN	O	O
-6	NN	O	O
)	NN	O	O
M	NN	O	O
induced	NN	O	O
rapid	NN	O	O
and	NN	O	O
significant	NN	O	O
elevation	NN	O	O
in	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
levels	NN	O	O
,	NN	O	O
whereas	NN	O	O
lyso-	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
was	NN	O	O
again	NN	O	O
ineffective	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
impact	NN	O	O
of	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
measuring	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
induced	NN	O	O
arachidonic	NN	O	O
acid	NN	O	O
release	NN	O	O
and	NN	O	O
5-hydroxyeicosatetraenoic	NN	O	O
acid	NN	O	O
production	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
was	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-DNA
proto-oncogenes	NN	O	I-DNA
c-fos	NN	O	B-DNA
and	NN	O	O
c-jun	NN	O	B-DNA
.	NN	O	O

Finally	NN	O	O
we	NN	O	O
explored	NN	O	O
the	NN	O	O
possible	NN	O	O
role	NN	O	O
of	NN	O	O
5-hydroxyeicosatetraenoic	NN	O	O
acid	NN	O	O
as	NN	O	O
a	NN	O	O
regulator	NN	O	O
of	NN	O	O
arachidonic	NN	O	O
acid	NN	O	O
liberation	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
endogenous	NN	O	O
5-lipoxygenase	NN	O	B-protein
activity	NN	O	O
modulates	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
induced	NN	O	O
arachidonic	NN	O	O
acid	NN	O	O
release	NN	O	O
perhaps	NN	O	O
acting	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
phospholipase	NN	O	B-protein
A2	NN	O	I-protein
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
platelet-activating	NN	O	B-protein
factor	NN	O	I-protein
is	NN	O	O
shown	NN	O	O
here	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
direct	NN	O	O
and	NN	O	O
profound	NN	O	O
effect	NN	O	O
on	NN	O	O
a	NN	O	O
pure	NN	O	B-cell_line
B	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Positive	NN	O	O
and	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
by	NN	O	O
a	NN	O	O
novel	NN	O	O
B-cell-specific	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
new	NN	O	O
B-cell-specific	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
identified	NN	O	O
3	NN	O	O
'	NN	O	O
of	NN	O	O
E4	NN	O	B-DNA
and	NN	O	I-DNA
the	NN	O	I-DNA
octamerlike	NN	O	I-DNA
motifs	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
heavy-chain	NN	O	I-DNA
gene	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Tandem	NN	O	O
copies	NN	O	O
of	NN	O	O
this	NN	O	O
67-bp	NN	O	B-DNA
MnlI-AluI	NN	O	I-DNA
fragment	NN	O	I-DNA
,	NN	O	O
when	NN	O	O
fused	NN	O	O
to	NN	O	O
the	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyltransferase	NN	O	I-DNA
gene	NN	O	I-DNA
driven	NN	O	O
by	NN	O	O
the	NN	O	O
conalbumin	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
stimulated	NN	O	O
transcription	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Footprinting	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
identical	NN	O	O
sequence	NN	O	O
CCGAAACTGAAAAGG	NN	O	O
,	NN	O	O
designated	NN	O	O
E6	NN	O	B-DNA
,	NN	O	O
was	NN	O	O
protected	NN	O	O
by	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
or	NN	O	O
HeLa	NN	O	O
cells	NN	O	O
.	NN	O	O

Gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
using	NN	O	O
a	NN	O	O
synthetic	NN	O	B-DNA
E6	NN	O	I-DNA
motif	NN	O	I-DNA
detected	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	B-protein
complex	NN	O	I-protein
in	NN	O	O
addition	NN	O	O
to	NN	O	O
a	NN	O	O
ubiquitous	NN	O	O
band	NN	O	O
found	NN	O	O
also	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
agreement	NN	O	O
with	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
gel	NN	O	O
retardation	NN	O	O
assays	NN	O	O
,	NN	O	O
tandem	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
E6	NN	O	B-DNA
motif	NN	O	I-DNA
stimulated	NN	O	O
transcription	NN	O	O
in	NN	O	O
ARH77	NN	O	B-cell_line
and	NN	O	O
Raji	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
or	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
a	NN	O	O
mutant	NN	O	B-DNA
E6	NN	O	I-DNA
motif	NN	O	I-DNA
lost	NN	O	O
both	NN	O	O
in	NN	O	O
vitro	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
in	NN	O	O
vivo	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
striking	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
mouse	NN	O	B-DNA
Ig	NN	O	I-DNA
heavy-chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
which	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
acts	NN	O	O
as	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
enhancer	NN	O	I-DNA
contains	NN	O	O
an	NN	O	O
octamerlike	NN	O	B-DNA
sequence	NN	O	I-DNA
with	NN	O	O
one	NN	O	O
base	NN	O	O
substitution	NN	O	O
which	NN	O	O
bound	NN	O	O
octamer-binding	NN	O	B-protein
proteins	NN	O	I-protein
with	NN	O	O
only	NN	O	O
very	NN	O	O
low	NN	O	O
affinity	NN	O	O
and	NN	O	O
showed	NN	O	O
no	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
of	NN	O	O
its	NN	O	O
own	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
MnlI-AluI	NN	O	B-DNA
fragment	NN	O	I-DNA
could	NN	O	O
suppress	NN	O	O
the	NN	O	O
basal-level	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
conalbumin	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
both	NN	O	O
Jurkat	NN	O	B-cell_line
and	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
simian	NN	O	O
virus	NN	O	O
40	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
was	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
MnlI-AluI	NN	O	B-DNA
fragment	NN	O	I-DNA
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
novel	NN	O	O
enhancer	NN	O	O
element	NN	O	O
identified	NN	O	O
in	NN	O	O
this	NN	O	O
study	NN	O	O
is	NN	O	O
probably	NN	O	O
a	NN	O	O
target	NN	O	O
site	NN	O	O
for	NN	O	O
both	NN	O	O
positive	NN	O	B-protein
and	NN	O	I-protein
negative	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Charybdotoxin-sensitive	NN	O	O
,	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
membrane	NN	O	O
potential	NN	O	O
changes	NN	O	O
are	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
human	NN	O	O
T	NN	O	B-cell_type
or	NN	O	I-cell_type
B	NN	O	I-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
and	NN	O	O
proliferation	NN	O	O
.	NN	O	O

The	NN	O	O
involvement	NN	O	O
of	NN	O	O
ion	NN	O	O
channels	NN	O	O
in	NN	O	O
B	NN	O	O
and	NN	O	O
T	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
is	NN	O	O
supported	NN	O	O
by	NN	O	O
many	NN	O	O
reports	NN	O	O
of	NN	O	O
changes	NN	O	O
in	NN	O	O
ion	NN	O	O
fluxes	NN	O	O
and	NN	O	O
membrane	NN	O	O
potential	NN	O	O
after	NN	O	O
mitogen	NN	O	B-protein
binding	NN	O	O
.	NN	O	O

Human	NN	O	B-cell_type
T	NN	O	I-cell_type
and	NN	O	I-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
demonstrate	NN	O	O
an	NN	O	O
early	NN	O	O
and	NN	O	O
transient	NN	O	O
hyperpolarization	NN	O	O
after	NN	O	O
ligand	NN	O	O
binding	NN	O	O
.	NN	O	O

Inasmuch	NN	O	O
as	NN	O	O
the	NN	O	O
change	NN	O	O
in	NN	O	O
membrane	NN	O	O
potential	NN	O	O
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
elevation	NN	O	O
of	NN	O	O
free	NN	O	O
cytosolic	NN	O	O
calcium	NN	O	O
,	NN	O	O
the	NN	O	O
hyperpolarization	NN	O	O
is	NN	O	O
presumably	NN	O	O
through	NN	O	O
opening	NN	O	O
of	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-stimulated	NN	O	O
K+	NN	O	O
channels	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
charybdotoxin	NN	O	O
,	NN	O	O
a	NN	O	O
known	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
Ca	NN	O	B-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-dependent	NN	O	I-protein
K+	NN	O	I-protein
channels	NN	O	I-protein
,	NN	O	O
to	NN	O	O
study	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
channels	NN	O	O
in	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
and	NN	O	O
mitogenesis	NN	O	O
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
charybdotoxin	NN	O	O
inhibits	NN	O	O
the	NN	O	O
ligand-induced	NN	O	O
transient	NN	O	O
membrane	NN	O	O
hyperpolarization	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
fashion	NN	O	O
,	NN	O	O
without	NN	O	O
affecting	NN	O	O
changes	NN	O	O
in	NN	O	O
cytosolic	NN	O	O
Ca2+	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
blockade	NN	O	O
of	NN	O	O
the	NN	O	O
Ca	NN	O	B-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-activated	NN	O	I-protein
K+	NN	O	I-protein
channel	NN	O	I-protein
is	NN	O	O
not	NN	O	O
associated	NN	O	O
with	NN	O	O
changes	NN	O	O
in	NN	O	O
cell-cycle	NN	O	O
gene	NN	O	O
activation	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
,	NN	O	O
IL-2R	NN	O	B-protein
expression	NN	O	O
or	NN	O	O
B	NN	O	O
and	NN	O	O
T	NN	O	O
cell	NN	O	O
mitogenesis	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
imply	NN	O	O
that	NN	O	O
membrane	NN	O	O
potential	NN	O	O
changes	NN	O	O
secondary	NN	O	O
to	NN	O	O
the	NN	O	O
ligand-dependent	NN	O	O
opening	NN	O	O
of	NN	O	O
Ca	NN	O	B-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-activated	NN	O	I-protein
K+	NN	O	I-protein
channels	NN	O	I-protein
are	NN	O	O
not	NN	O	O
involved	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
and	NN	O	I-cell_type
T	NN	O	I-cell_type
lymphocyte	NN	O	I-cell_type
activation	NN	O	O
and	NN	O	O
mitogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activity	NN	O	O
of	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
in	NN	O	O
somatic	NN	O	B-cell_line
cell	NN	O	I-cell_line
hybrids	NN	O	I-cell_line
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
the	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

The	NN	O	O
two	NN	O	B-protein
nuclear	NN	O	I-protein
proteins	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
consisting	NN	O	O
of	NN	O	O
subunits	NN	O	O
p50	NN	O	B-protein
an	NN	O	O
dp65	NN	O	B-protein
)	NN	O	O
and	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
p50	NN	O	B-protein
)	NN	O	O
by	NN	O	O
itself	NN	O	O
,	NN	O	O
also	NN	O	O
called	NN	O	O
KBF1	NN	O	B-protein
,	NN	O	O
are	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
and	NN	O	O
localized	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
IARC	NN	O	B-cell_line
301.5	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
roles	NN	O	O
of	NN	O	O
these	NN	O	O
two	NN	O	O
factors	NN	O	B-protein
,	NN	O	O
which	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
same	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancers	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
transcription	NN	O	O
activation	NN	O	O
we	NN	O	O
have	NN	O	O
prepared	NN	O	O
somatic	NN	O	O
cell	NN	O	O
hybrids	NN	O	O
between	NN	O	O
IARC	NN	O	B-cell_line
301.5	NN	O	I-cell_line
and	NN	O	O
a	NN	O	O
murine	NN	O	B-cell_line
myeloma	NN	O	I-cell_line
.	NN	O	O

Most	NN	O	O
hybrids	NN	O	O
express	NN	O	O
both	NN	O	O
KBF1	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
their	NN	O	O
nuclei	NN	O	O
,	NN	O	O
but	NN	O	O
one	NN	O	O
hybrid	NN	O	O
expresses	NN	O	O
only	NN	O	O
KBF1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	I-protein
IL-2	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
chain	NN	O	I-protein
(	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
is	NN	O	O
functional	NN	O	O
only	NN	O	O
in	NN	O	O
the	NN	O	O
hybrids	NN	O	B-cell_line
expressing	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

These	NN	O	O
findings	NN	O	O
show	NN	O	O
that	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
necessary	NN	O	O
to	NN	O	O
activate	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
while	NN	O	O
KBF1	NN	O	B-protein
by	NN	O	O
itself	NN	O	O
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
KBF1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
competitive	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
discuss	NN	O	O
how	NN	O	O
these	NN	O	O
factors	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
transient	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
alpha	NN	O	I-protein
genes	NN	O	O
during	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

T-helper-cell	NN	O	B-protein
determinants	NN	O	I-protein
in	NN	O	O
protein	NN	O	B-protein
antigens	NN	O	I-protein
are	NN	O	O
preferentially	NN	O	O
located	NN	O	O
in	NN	O	O
cysteine-rich	NN	O	B-protein
antigen	NN	O	I-protein
segments	NN	O	I-protein
resistant	NN	O	O
to	NN	O	O
proteolytic	NN	O	O
cleavage	NN	O	O
by	NN	O	O
cathepsin	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	I-protein
L	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
D	NN	O	I-protein
.	NN	O	O

We	NN	O	O
report	NN	O	O
on	NN	O	O
a	NN	O	O
computer	NN	O	O
algorithm	NN	O	O
capable	NN	O	O
of	NN	O	O
predicting	NN	O	O
the	NN	O	O
location	NN	O	O
of	NN	O	O
T-helper-cell	NN	O	B-protein
epitopes	NN	O	I-protein
in	NN	O	O
protein	NN	O	B-protein
antigen	NN	O	I-protein
(	NN	O	O
Ag	NN	O	B-protein
)	NN	O	O
by	NN	O	O
analysing	NN	O	O
the	NN	O	O
Ag	NN	O	B-protein
amino	NN	O	B-protein
acid	NN	O	I-protein
sequence	NN	O	I-protein
.	NN	O	O

The	NN	O	O
algorithm	NN	O	O
was	NN	O	O
constructed	NN	O	O
with	NN	O	O
the	NN	O	O
aim	NN	O	O
of	NN	O	O
identifying	NN	O	O
segments	NN	O	O
in	NN	O	O
Ag	NN	O	B-protein
which	NN	O	O
are	NN	O	O
resistant	NN	O	O
to	NN	O	O
proteolytic	NN	O	O
degradation	NN	O	O
by	NN	O	O
the	NN	O	O
enzymes	NN	O	O
cathepsin	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	I-protein
L	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
D	NN	O	I-protein
.	NN	O	O

These	NN	O	O
are	NN	O	O
prominent	NN	O	O
enzymes	NN	O	B-protein
in	NN	O	O
the	NN	O	O
endocytic	NN	O	O
pathway	NN	O	O
through	NN	O	O
which	NN	O	O
soluble	NN	O	B-protein
protein	NN	O	I-protein
Ag	NN	O	I-protein
enter	NN	O	O
APC	NN	O	B-protein
,	NN	O	O
and	NN	O	O
resistant	NN	O	B-protein
segments	NN	O	I-protein
in	NN	O	O
Ag	NN	O	B-protein
may	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
be	NN	O	O
expected	NN	O	O
to	NN	O	O
contain	NN	O	O
more	NN	O	O
T-cell	NN	O	B-protein
determinants	NN	O	I-protein
than	NN	O	O
susceptible	NN	O	B-protein
segments	NN	O	I-protein
.	NN	O	O

From	NN	O	O
information	NN	O	O
available	NN	O	O
in	NN	O	O
the	NN	O	O
literature	NN	O	O
on	NN	O	O
the	NN	O	O
substrate	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
enzymes	NN	O	B-protein
,	NN	O	O
it	NN	O	O
is	NN	O	O
clear	NN	O	O
that	NN	O	O
a	NN	O	O
cysteine	NN	O	O
is	NN	O	O
not	NN	O	O
accepted	NN	O	O
in	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
S2	NN	O	B-protein
,	NN	O	I-protein
S1	NN	O	I-protein
,	NN	O	I-protein
S1	NN	O	I-protein
'	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
S2	NN	O	I-protein
'	NN	O	I-protein
subsites	NN	O	I-protein
of	NN	O	O
cathepsin	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	I-protein
L	NN	O	I-protein
,	NN	O	O
and	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
S1	NN	O	B-protein
and	NN	O	O
S1	NN	O	B-protein
'	NN	O	I-protein
subsites	NN	O	O
of	NN	O	O
cathepsin	NN	O	B-protein
D	NN	O	I-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
noticed	NN	O	O
that	NN	O	O
cysteine-containing	NN	O	B-protein
T-cell	NN	O	I-protein
determinants	NN	O	I-protein
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
Ag	NN	O	I-protein
are	NN	O	O
particularly	NN	O	O
rich	NN	O	O
in	NN	O	O
the	NN	O	O
amino	NN	O	O
acids	NN	O	O
alanine	NN	O	O
,	NN	O	O
glycine	NN	O	O
,	NN	O	O
lysine	NN	O	O
,	NN	O	O
leucine	NN	O	O
,	NN	O	O
serine	NN	O	O
,	NN	O	O
threonine	NN	O	O
,	NN	O	O
and	NN	O	O
valine	NN	O	O
.	NN	O	O

By	NN	O	O
searching	NN	O	O
protein	NN	O	O
Ag	NN	O	B-protein
for	NN	O	O
clusters	NN	O	O
of	NN	O	O
amino	NN	O	O
acids	NN	O	O
containing	NN	O	O
cysteine	NN	O	O
and	NN	O	O
two	NN	O	O
of	NN	O	O
the	NN	O	O
other	NN	O	O
amino	NN	O	O
acids	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
predict	NN	O	O
17	NN	O	O
out	NN	O	O
of	NN	O	O
23	NN	O	O
empirically	NN	O	O
known	NN	O	O
T-cell	NN	O	B-protein
determinants	NN	O	I-protein
in	NN	O	O
the	NN	O	O
Ag	NN	O	B-protein
with	NN	O	O
a	NN	O	O
relatively	NN	O	O
low	NN	O	O
number	NN	O	O
of	NN	O	O
false	NN	O	O
(	NN	O	O
positive	NN	O	O
)	NN	O	O
predictions	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
present	NN	O	O
a	NN	O	O
new	NN	O	O
principle	NN	O	O
for	NN	O	O
searching	NN	O	O
Ag	NN	O	B-protein
for	NN	O	O
potential	NN	O	O
amphipatic	NN	O	B-protein
alpha-helical	NN	O	I-protein
protein	NN	O	I-protein
segments	NN	O	I-protein
.	NN	O	O

Such	NN	O	O
segments	NN	O	O
accord	NN	O	O
well	NN	O	O
with	NN	O	O
empirically	NN	O	O
known	NN	O	O
T-cell	NN	O	B-protein
determinants	NN	O	I-protein
and	NN	O	O
our	NN	O	O
algorithm	NN	O	O
produces	NN	O	O
a	NN	O	O
lower	NN	O	O
number	NN	O	O
of	NN	O	O
false	NN	O	O
positive	NN	O	O
predictions	NN	O	O
than	NN	O	O
the	NN	O	O
principle	NN	O	O
based	NN	O	O
on	NN	O	O
discrete	NN	O	O
Fourier	NN	O	O
transformations	NN	O	O
previously	NN	O	O
described	NN	O	O
.	NN	O	O

-DOCSTART-	O

Contribution	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
Sp1	NN	O	B-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
to	NN	O	O
the	NN	O	O
replicative	NN	O	O
capacity	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
:	NN	O	O
distinct	NN	O	O
patterns	NN	O	O
of	NN	O	O
viral	NN	O	O
growth	NN	O	O
are	NN	O	O
determined	NN	O	O
by	NN	O	O
T-cell	NN	O	B-cell_type
types	NN	O	I-cell_type
.	NN	O	O

Starting	NN	O	O
with	NN	O	O
a	NN	O	O
replication-incompetent	NN	O	O
molecular	NN	O	O
clone	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
,	NN	O	O
lacking	NN	O	O
all	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
Sp1	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
the	NN	O	O
native	NN	O	B-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
proviruses	NN	O	O
containing	NN	O	O
reconstructed	NN	O	B-DNA
LTRs	NN	O	I-DNA
with	NN	O	O
individual	NN	O	O
or	NN	O	O
combinations	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
and	NN	O	I-DNA
Sp1	NN	O	I-DNA
elements	NN	O	I-DNA
were	NN	O	O
generated	NN	O	O
and	NN	O	O
evaluated	NN	O	O
for	NN	O	O
their	NN	O	O
capacity	NN	O	O
to	NN	O	O
produce	NN	O	O
virus	NN	O	O
progeny	NN	O	O
following	NN	O	O
transfection-cocultivation	NN	O	O
.	NN	O	O

Virus	NN	O	O
stocks	NN	O	O
obtained	NN	O	O
from	NN	O	O
these	NN	O	O
experiments	NN	O	O
exhibited	NN	O	O
a	NN	O	O
continuum	NN	O	O
of	NN	O	O
replicative	NN	O	O
capacities	NN	O	O
in	NN	O	O
different	NN	O	O
human	NN	O	B-cell_type
T-cell	NN	O	I-cell_type
types	NN	O	I-cell_type
depending	NN	O	O
on	NN	O	O
which	NN	O	O
element	NN	O	B-DNA
(	NN	O	O
s	NN	O	O
)	NN	O	O
was	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
LTR	NN	O	B-DNA
.	NN	O	O

For	NN	O	O
example	NN	O	O
,	NN	O	O
in	NN	O	O
experiments	NN	O	O
involving	NN	O	O
proviral	NN	O	O
clones	NN	O	O
with	NN	O	O
LTRs	NN	O	B-DNA
containing	NN	O	O
one	NN	O	O
or	NN	O	O
two	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
elements	NN	O	I-DNA
(	NN	O	O
and	NN	O	O
no	NN	O	O
Sp1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
a	NN	O	O
hierarchy	NN	O	O
of	NN	O	O
cellular	NN	O	O
permissivity	NN	O	O
to	NN	O	O
virus	NN	O	O
replication	NN	O	O
(	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
=	NN	O	O
MT4	NN	O	B-cell_line
greater	NN	O	O
than	NN	O	O
H9	NN	O	B-cell_line
greater	NN	O	O
than	NN	O	O
CEM	NN	O	B-cell_line
greater	NN	O	O
than	NN	O	O
Jurkat	NN	O	B-cell_line
)	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

Of	NN	O	O
note	NN	O	O
was	NN	O	O
the	NN	O	O
associated	NN	O	O
emergence	NN	O	O
of	NN	O	O
second-site	NN	O	B-DNA
LTR	NN	O	I-DNA
revertants	NN	O	O
which	NN	O	O
involved	NN	O	O
an	NN	O	O
alteration	NN	O	O
of	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1	NN	O	I-DNA
LTR	NN	O	I-DNA
possesses	NN	O	O
functional	NN	O	O
redundancy	NN	O	O
which	NN	O	O
ensures	NN	O	O
virus	NN	O	O
replication	NN	O	O
in	NN	O	O
different	NN	O	O
T-cell	NN	O	B-cell_type
types	NN	O	I-cell_type
and	NN	O	O
is	NN	O	O
capable	NN	O	O
of	NN	O	O
changing	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
particular	NN	O	O
combination	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
factors	NN	O	I-protein
present	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stimulation	NN	O	O
of	NN	O	O
interferon	NN	O	B-DNA
beta	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
purified	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
a	NN	O	O
novel	NN	O	O
TH	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
human	NN	O	B-DNA
interferon	NN	O	I-DNA
beta	NN	O	I-DNA
(	NN	O	I-DNA
IFN-beta	NN	O	I-DNA
)	NN	O	I-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
consists	NN	O	O
of	NN	O	O
multiple	NN	O	O
enhanson	NN	O	B-DNA
domains	NN	O	I-DNA
which	NN	O	O
are	NN	O	O
targets	NN	O	O
for	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

To	NN	O	O
further	NN	O	O
characterize	NN	O	O
the	NN	O	O
protein-DNA	NN	O	O
interactions	NN	O	O
mediating	NN	O	O
IFN-beta	NN	O	B-protein
induction	NN	O	O
,	NN	O	O
positive	NN	O	B-protein
regulatory	NN	O	I-protein
domain	NN	O	I-protein
(	NN	O	I-protein
PRD	NN	O	I-protein
)	NN	O	I-protein
II	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
were	NN	O	O
purified	NN	O	O
from	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
induced	NN	O	O
Jurkat	NN	O	B-cell_line
T-cells	NN	O	I-cell_line
and	NN	O	O
from	NN	O	O
IFN	NN	O	B-cell_line
primed	NN	O	I-cell_line
,	NN	O	I-cell_line
cycloheximide/polyinosinic-polycytidylic	NN	O	I-cell_line
acid	NN	O	I-cell_line
treated	NN	O	I-cell_line
HeLa	NN	O	I-cell_line
S3	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

From	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
two	NN	O	O
major	NN	O	O
proteins	NN	O	O
of	NN	O	O
52	NN	O	B-protein
and	NN	O	I-protein
45	NN	O	I-protein
kilodaltons	NN	O	I-protein
(	NN	O	I-protein
kD	NN	O	I-protein
)	NN	O	I-protein
copurified	NN	O	O
with	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
whereas	NN	O	O
from	NN	O	O
T-cells	NN	O	B-cell_type
,	NN	O	O
four	NN	O	O
proteins	NN	O	O
--	NN	O	O
a	NN	O	O
major	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
52	NN	O	B-protein
kD	NN	O	I-protein
and	NN	O	O
three	NN	O	O
minor	NN	O	B-protein
proteins	NN	O	I-protein
of	NN	O	O
82	NN	O	B-protein
,	NN	O	O
67	NN	O	B-protein
,	NN	O	O
and	NN	O	O
43-47	NN	O	B-protein
kD	NN	O	I-protein
--	NN	O	O
were	NN	O	O
purified	NN	O	O
.	NN	O	O

Also	NN	O	O
,	NN	O	O
an	NN	O	O
induction	NN	O	O
specific	NN	O	B-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
purified	NN	O	O
from	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
that	NN	O	O
interacted	NN	O	O
with	NN	O	O
the	NN	O	O
(	NN	O	O
AAGTGA	NN	O	O
)	NN	O	O
4	NN	O	O
tetrahexamer	NN	O	B-DNA
sequence	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
PRDI	NN	O	B-DNA
domain	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
protein	NN	O	O
is	NN	O	O
immunologically	NN	O	O
distinct	NN	O	O
from	NN	O	O
IRF-1/ISGF2	NN	O	B-protein
.	NN	O	O

Uninduced	NN	O	O
or	NN	O	O
Sendai	NN	O	O
virus	NN	O	O
induced	NN	O	O
HeLa	NN	O	O
extracts	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
examine	NN	O	O
transcription	NN	O	O
in	NN	O	O
vitro	NN	O	O
using	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
IFN	NN	O	B-DNA
beta	NN	O	I-DNA
promoter	NN	O	I-DNA
deletions	NN	O	I-DNA
.	NN	O	O

Deletions	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
PRDII	NN	O	B-DNA
element	NN	O	I-DNA
increased	NN	O	O
transcription	NN	O	O
in	NN	O	O
the	NN	O	O
uninduced	NN	O	O
extract	NN	O	O
,	NN	O	O
indicating	NN	O	O
predominantly	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

A	NN	O	O
2-4-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
IFN-beta	NN	O	B-DNA
promoter	NN	O	I-DNA
transcription	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
Sendai	NN	O	O
virus	NN	O	O
induced	NN	O	O
extracts	NN	O	O
,	NN	O	O
and	NN	O	O
deletion	NN	O	O
of	NN	O	O
PRDI	NN	O	B-DNA
and	NN	O	O
PRDII	NN	O	B-DNA
elements	NN	O	I-DNA
decreased	NN	O	O
this	NN	O	O
induced	NN	O	O
level	NN	O	O
of	NN	O	O
transcription	NN	O	O
.	NN	O	O

When	NN	O	O
purified	NN	O	O
PRDII	NN	O	B-protein
and	NN	O	I-protein
tetrahexamer	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
were	NN	O	O
added	NN	O	O
to	NN	O	O
the	NN	O	O
induced	NN	O	O
extract	NN	O	O
,	NN	O	O
a	NN	O	O
4-fold	NN	O	O
increase	NN	O	O
in	NN	O	O
transcription	NN	O	O
was	NN	O	O
observed	NN	O	O
.	NN	O	O

These	NN	O	O
experiments	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
possible	NN	O	O
to	NN	O	O
modulate	NN	O	O
IFN-beta	NN	O	O
transcription	NN	O	O
in	NN	O	O
vitro	NN	O	O
but	NN	O	O
indicate	NN	O	O
that	NN	O	O
additional	NN	O	O
proteins	NN	O	O
may	NN	O	O
be	NN	O	O
required	NN	O	O
to	NN	O	O
fully	NN	O	O
activate	NN	O	O
IFN-beta	NN	O	B-protein
transcription	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
rhombotin	NN	O	B-DNA
family	NN	O	I-DNA
of	NN	O	O
cysteine-rich	NN	O	B-DNA
LIM-domain	NN	O	I-DNA
oncogenes	NN	O	I-DNA
:	NN	O	O
distinct	NN	O	O
members	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
T-cell	NN	O	B-DNA
translocations	NN	O	I-DNA
to	NN	O	O
human	NN	O	B-DNA
chromosomes	NN	O	I-DNA
11p15	NN	O	I-DNA
and	NN	O	I-DNA
11p13	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
chromosomal	NN	O	O
translocation	NN	O	O
in	NN	O	O
a	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
involving	NN	O	O
the	NN	O	O
short	NN	O	B-DNA
arm	NN	O	I-DNA
of	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
11	NN	O	I-DNA
at	NN	O	O
band	NN	O	B-DNA
11p15	NN	O	I-DNA
disrupts	NN	O	O
the	NN	O	O
rhombotin	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
gene	NN	O	O
encodes	NN	O	O
a	NN	O	O
protein	NN	O	O
with	NN	O	O
duplicated	NN	O	O
cysteine-rich	NN	O	B-protein
regions	NN	O	I-protein
called	NN	O	O
LIM	NN	O	B-protein
domains	NN	O	I-protein
,	NN	O	O
which	NN	O	O
show	NN	O	O
homology	NN	O	O
to	NN	O	O
zinc-binding	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
to	NN	O	O
iron-sulfur	NN	O	B-protein
centers	NN	O	I-protein
of	NN	O	O
ferredoxins	NN	O	B-protein
.	NN	O	O

Two	NN	O	O
homologues	NN	O	O
of	NN	O	O
the	NN	O	O
rhombotin	NN	O	B-DNA
gene	NN	O	I-DNA
have	NN	O	O
now	NN	O	O
been	NN	O	O
isolated	NN	O	O
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
,	NN	O	O
designated	NN	O	O
Rhom-2	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
located	NN	O	O
on	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
11	NN	O	I-DNA
at	NN	O	O
band	NN	O	B-DNA
11p13	NN	O	I-DNA
,	NN	O	O
where	NN	O	O
a	NN	O	O
cluster	NN	O	O
of	NN	O	O
T-cell	NN	O	B-DNA
leukemia-specific	NN	O	I-DNA
translocations	NN	O	I-DNA
occur	NN	O	O
;	NN	O	O
all	NN	O	O
translocation	NN	O	O
breakpoints	NN	O	O
at	NN	O	O
11p13	NN	O	B-DNA
are	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
Rhom-2	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Human	NN	O	B-DNA
and	NN	O	I-DNA
mouse	NN	O	I-DNA
Rhom-2	NN	O	I-DNA
are	NN	O	O
highly	NN	O	O
conserved	NN	O	O
and	NN	O	O
,	NN	O	O
like	NN	O	O
rhombotin	NN	O	B-DNA
,	NN	O	O
encode	NN	O	O
two	NN	O	O
tandem	NN	O	B-DNA
cysteine-rich	NN	O	I-DNA
LIM	NN	O	I-DNA
domains	NN	O	I-DNA
.	NN	O	O

Rhom-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
expressed	NN	O	O
in	NN	O	O
early	NN	O	O
mouse	NN	O	O
development	NN	O	O
in	NN	O	O
central	NN	O	O
nervous	NN	O	O
system	NN	O	O
,	NN	O	O
lung	NN	O	O
,	NN	O	O
kidney	NN	O	O
,	NN	O	O
liver	NN	O	O
,	NN	O	O
and	NN	O	O
spleen	NN	O	O
but	NN	O	O
only	NN	O	O
very	NN	O	O
low	NN	O	O
levels	NN	O	O
occur	NN	O	O
in	NN	O	O
thymus	NN	O	O
.	NN	O	O

The	NN	O	O
other	NN	O	O
gene	NN	O	O
,	NN	O	O
designated	NN	O	O
Rhom-3	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
not	NN	O	O
on	NN	O	O
chromosome	NN	O	O
11	NN	O	O
but	NN	O	O
also	NN	O	O
retains	NN	O	O
homology	NN	O	O
to	NN	O	O
the	NN	O	O
LIM	NN	O	B-DNA
domain	NN	O	I-DNA
of	NN	O	O
rhombotin	NN	O	B-DNA
.	NN	O	O

Since	NN	O	O
the	NN	O	O
Rhom-2	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
such	NN	O	O
a	NN	O	O
common	NN	O	O
site	NN	O	O
of	NN	O	O
chromosomal	NN	O	O
damage	NN	O	O
in	NN	O	O
T-cell	NN	O	B-cell_type
tumors	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
consistency	NN	O	O
of	NN	O	O
translocations	NN	O	O
near	NN	O	O
the	NN	O	O
rhombotin	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
further	NN	O	O
examined	NN	O	O
.	NN	O	O

A	NN	O	O
second	NN	O	O
translocation	NN	O	O
adjacent	NN	O	O
to	NN	O	O
rhombotin	NN	O	B-DNA
was	NN	O	O
found	NN	O	O
and	NN	O	O
at	NN	O	O
the	NN	O	O
same	NN	O	O
position	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
previous	NN	O	O
example	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
chromosome	NN	O	B-DNA
bands	NN	O	I-DNA
11p15	NN	O	I-DNA
(	NN	O	O
rhombotin	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
11p13	NN	O	B-DNA
(	NN	O	O
Rhom-2	NN	O	B-DNA
)	NN	O	O
are	NN	O	O
consistent	NN	O	O
sites	NN	O	O
of	NN	O	O
chromosome	NN	O	O
translocation	NN	O	O
in	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
11p15	NN	O	B-DNA
target	NN	O	I-DNA
more	NN	O	O
rarely	NN	O	O
involved	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
define	NN	O	O
the	NN	O	O
rhombotin	NN	O	B-DNA
gene	NN	O	O
family	NN	O	O
as	NN	O	O
a	NN	O	O
class	NN	O	O
of	NN	O	O
T-cell	NN	O	B-DNA
oncogenes	NN	O	I-DNA
with	NN	O	O
duplicated	NN	O	O
cysteine-rich	NN	O	B-DNA
LIM	NN	O	I-DNA
domains	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
in	NN	O	O
the	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
is	NN	O	O
independent	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
,	NN	O	O
and	NN	O	O
Ca	NN	O	B-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-regulated	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
able	NN	O	O
to	NN	O	O
control	NN	O	O
transcription	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
genes	NN	O	O
,	NN	O	O
including	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
(	NN	O	I-DNA
HIV	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
activated	NN	O	O
upon	NN	O	O
cellular	NN	O	O
treatment	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
and	NN	O	O
the	NN	O	O
cytokine	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
(	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
work	NN	O	O
,	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
molecular	NN	O	O
events	NN	O	O
leading	NN	O	O
to	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
Jurkat	NN	O	B-cell_line
)	NN	O	O
and	NN	O	O
its	NN	O	O
subclone	NN	O	O
JCT6	NN	O	B-cell_line
,	NN	O	O
which	NN	O	O
presents	NN	O	O
a	NN	O	O
deficiency	NN	O	O
in	NN	O	O
the	NN	O	O
PKA	NN	O	B-protein
transduction	NN	O	O
pathway	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
in	NN	O	O
both	NN	O	O
cell	NN	O	O
lines	NN	O	O
,	NN	O	O
both	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
and	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
were	NN	O	O
able	NN	O	O
to	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Phorbol	NN	O	O
activation	NN	O	O
was	NN	O	O
positively	NN	O	O
modulated	NN	O	O
by	NN	O	O
Ca2+	NN	O	O
influx	NN	O	O
while	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
not	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
while	NN	O	O
PMA	NN	O	O
activation	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
PKC	NN	O	B-protein
inhibitor	NN	O	O
staurosporin	NN	O	O
,	NN	O	O
the	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
effect	NN	O	O
was	NN	O	O
unchanged	NN	O	O
.	NN	O	O

TNF	NN	O	B-protein
alpha	NN	O	I-protein
did	NN	O	O
not	NN	O	O
activate	NN	O	O
cAMP	NN	O	O
production	NN	O	O
and	NN	O	O
its	NN	O	O
signal	NN	O	O
was	NN	O	O
not	NN	O	O
modulated	NN	O	O
by	NN	O	O
cAMP	NN	O	O
activators	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
cAMP	NN	O	O
activators	NN	O	O
did	NN	O	O
not	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
major	NN	O	O
signal-mediating	NN	O	B-protein
kinases	NN	O	I-protein
such	NN	O	O
as	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
(	NN	O	O
PKC	NN	O	B-protein
)	NN	O	O
,	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
A	NN	O	I-protein
,	NN	O	O
or	NN	O	O
Ca	NN	O	B-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-regulated	NN	O	I-protein
kinases	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
cytoplasmic	NN	O	O
acidification	NN	O	O
facilitated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
both	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
PKC	NN	O	B-protein
,	NN	O	O
by	NN	O	O
a	NN	O	O
mechanism	NN	O	O
that	NN	O	O
increases	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
/I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
dissociation	NN	O	O
without	NN	O	O
affecting	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
step	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
functional	NN	O	O
domains	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
Thy-1	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
Thy-1	NN	O	B-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
resembles	NN	O	O
a	NN	O	O
``	NN	O	B-DNA
housekeeping	NN	O	I-DNA
''	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
located	NN	O	O
within	NN	O	O
a	NN	O	O
methylation-free	NN	O	B-DNA
island	NN	O	I-DNA
,	NN	O	O
lacks	NN	O	O
a	NN	O	O
canonical	NN	O	B-DNA
TATA	NN	O	I-DNA
box	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
displays	NN	O	O
heterogeneity	NN	O	O
in	NN	O	O
the	NN	O	O
5'-end	NN	O	B-RNA
termini	NN	O	I-RNA
of	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
.	NN	O	O

Using	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
does	NN	O	O
not	NN	O	O
confer	NN	O	O
any	NN	O	O
tissue	NN	O	O
specificity	NN	O	O
and	NN	O	O
is	NN	O	O
active	NN	O	O
only	NN	O	O
in	NN	O	O
a	NN	O	O
position-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

It	NN	O	O
can	NN	O	O
only	NN	O	O
be	NN	O	O
activated	NN	O	O
in	NN	O	O
a	NN	O	O
tissue-specific	NN	O	O
manner	NN	O	O
by	NN	O	O
elements	NN	O	O
that	NN	O	O
lie	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
initiation	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
functional	NN	O	O
domains	NN	O	O
of	NN	O	O
the	NN	O	O
minimal	NN	O	B-DNA
Thy-1	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
dominant	NN	O	B-DNA
promoter	NN	O	I-DNA
elements	NN	O	I-DNA
consist	NN	O	O
of	NN	O	O
multiple	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Sp1	NN	O	B-protein
,	NN	O	O
an	NN	O	O
inverted	NN	O	O
CCAAT	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
sequences	NN	O	O
proximal	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
start	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

DNase	NN	O	O
I	NN	O	O
and	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
show	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
these	NN	O	O
elements	NN	O	B-DNA
,	NN	O	O
including	NN	O	O
Sp1	NN	O	B-protein
and	NN	O	O
CP1	NN	O	B-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
structure	NN	O	O
of	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
only	NN	O	O
permits	NN	O	O
productive	NN	O	O
interactions	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
Sp1	NN	O	B-protein
and	NN	O	O
CP1	NN	O	B-protein
with	NN	O	O
the	NN	O	O
basal	NN	O	O
transcription	NN	O	O
machinery	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
enhancer	NN	O	B-DNA
sequences	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activates	NN	O	O
proenkephalin	NN	O	O
transcription	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Upon	NN	O	O
activation	NN	O	O
,	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
accumulate	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
neuropeptide	NN	O	O
enkephalin	NN	O	O
which	NN	O	O
correlate	NN	O	O
with	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
proenkephalin	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
investigated	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
basis	NN	O	O
for	NN	O	O
these	NN	O	O
changes	NN	O	O
.	NN	O	O

The	NN	O	O
proenkephalin	NN	O	O
promoter	NN	O	O
contains	NN	O	O
a	NN	O	O
sequence	NN	O	O
GGGGACGTCCCC	NN	O	O
,	NN	O	O
named	NN	O	O
B2	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
sequence	NN	O	I-DNA
GGGGACTTTCC	NN	O	O
,	NN	O	O
the	NN	O	O
binding	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
induces	NN	O	O
an	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-like	NN	O	O
binding	NN	O	O
activity	NN	O	O
to	NN	O	O
the	NN	O	O
B2	NN	O	B-DNA
site	NN	O	O
,	NN	O	O
concomitant	NN	O	O
with	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
proenkephalin	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Mutations	NN	O	O
at	NN	O	O
the	NN	O	O
B2	NN	O	B-DNA
site	NN	O	O
abolish	NN	O	O
this	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
purified	NN	O	O
homodimer	NN	O	B-protein
(	NN	O	O
two	NN	O	O
p50s	NN	O	B-protein
)	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binds	NN	O	O
the	NN	O	O
B2	NN	O	B-DNA
site	NN	O	O
of	NN	O	O
proenkephalin	NN	O	O
relatively	NN	O	O
better	NN	O	O
than	NN	O	O
does	NN	O	O
the	NN	O	O
heterotetramer	NN	O	B-protein
(	NN	O	O
two	NN	O	O
p65s	NN	O	B-protein
plus	NN	O	O
two	NN	O	O
p50s	NN	O	B-protein
)	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
factor	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
it	NN	O	O
appears	NN	O	O
that	NN	O	O
the	NN	O	O
T-cell-specific	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
proenkephalin	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
as	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
ubiquitous	NN	O	O
and	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
through	NN	O	O
the	NN	O	O
B2	NN	O	B-DNA
site	NN	O	O
is	NN	O	O
T	NN	O	O
cell	NN	O	O
specific	NN	O	O
,	NN	O	O
yet	NN	O	O
another	NN	O	O
T-cell-specific	NN	O	B-protein
factor	NN	O	I-protein
which	NN	O	O
synergizes	NN	O	O
with	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
should	NN	O	O
be	NN	O	O
considered	NN	O	O
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
NF-KB	NN	O	B-protein
during	NN	O	O
monocyte	NN	O	B-cell_type
differentiation	NN	O	O
by	NN	O	O
HIV	NN	O	O
type	NN	O	O
1	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
production	NN	O	O
of	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
progeny	NN	O	O
was	NN	O	O
followed	NN	O	O
in	NN	O	O
the	NN	O	O
U937	NN	O	B-cell_line
promonocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
after	NN	O	O
stimulation	NN	O	O
either	NN	O	O
with	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
or	NN	O	O
PMA	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
purified	NN	O	B-cell_type
human	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	I-cell_type
macrophages	NN	O	I-cell_type
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
and	NN	O	O
Southwestern	NN	O	O
blotting	NN	O	O
experiments	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
detect	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
transactivation	NN	O	I-protein
factor	NN	O	I-protein
NF-KB	NN	O	B-protein
to	NN	O	O
the	NN	O	O
double	NN	O	B-DNA
repeat-KB	NN	O	I-DNA
enhancer	NN	O	I-DNA
sequence	NN	O	I-DNA
located	NN	O	O
in	NN	O	O
the	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

PMA	NN	O	O
treatment	NN	O	O
,	NN	O	O
and	NN	O	O
not	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
acts	NN	O	O
in	NN	O	O
inducing	NN	O	O
NF-KB	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
the	NN	O	O
nuclei	NN	O	O
.	NN	O	O

In	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
monocytes	NN	O	B-cell_type
or	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-KB	NN	O	B-protein
occurred	NN	O	O
only	NN	O	O
if	NN	O	O
the	NN	O	O
cells	NN	O	O
were	NN	O	O
previously	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

When	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
no	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-KB	NN	O	B-protein
factor	NN	O	I-protein
was	NN	O	O
detected	NN	O	O
,	NN	O	O
whereas	NN	O	O
high	NN	O	O
level	NN	O	O
of	NN	O	O
progeny	NN	O	O
virions	NN	O	O
was	NN	O	O
produced	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
factor	NN	O	O
was	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
viral	NN	O	O
replication	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lineage	NN	O	I-cell_line
,	NN	O	O
HIV-1	NN	O	O
could	NN	O	O
mimic	NN	O	O
some	NN	O	O
differentiation/activation	NN	O	O
stimuli	NN	O	O
allowing	NN	O	O
nuclear	NN	O	O
NF-KB	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

