-DOCSTART-	O

Disruption	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
SCL	NN	O	I-DNA
locus	NN	O	I-DNA
by	NN	O	O
``	NN	O	O
illegitimate	NN	O	O
''	NN	O	O
V-	NN	O	O
(	NN	O	O
D	NN	O	O
)	NN	O	O
-J	NN	O	O
recombinase	NN	O	O
activity	NN	O	O
.	NN	O	O

A	NN	O	O
fusion	NN	O	B-DNA
complementary	NN	O	I-DNA
DNA	NN	O	I-DNA
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
HSB-2	NN	O	I-cell_line
elucidates	NN	O	O
a	NN	O	O
provocative	NN	O	O
mechanism	NN	O	O
for	NN	O	O
the	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
putative	NN	O	O
hematopoietic	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
SCL	NN	O	B-protein
.	NN	O	O

The	NN	O	O
fusion	NN	O	B-DNA
cDNA	NN	O	I-DNA
results	NN	O	O
from	NN	O	O
an	NN	O	O
interstitial	NN	O	O
deletion	NN	O	O
between	NN	O	O
a	NN	O	O
previously	NN	O	O
unknown	NN	O	B-DNA
locus	NN	O	I-DNA
,	NN	O	O
SIL	NN	O	B-DNA
(	NN	O	O
SCL	NN	O	B-DNA
interrupting	NN	O	I-DNA
locus	NN	O	I-DNA
)	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
untranslated	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
SCL	NN	O	B-protein
.	NN	O	O

Similar	NN	O	O
to	NN	O	O
1	NN	O	O
;	NN	O	O
14	NN	O	O
translocations	NN	O	O
,	NN	O	O
this	NN	O	O
deletion	NN	O	O
disrupts	NN	O	O
the	NN	O	O
SCL	NN	O	B-DNA
5	NN	O	I-DNA
'	NN	O	I-DNA
regulatory	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
event	NN	O	O
is	NN	O	O
probably	NN	O	O
mediated	NN	O	O
by	NN	O	O
V-	NN	O	O
(	NN	O	O
D	NN	O	O
)	NN	O	O
-J	NN	O	O
recombinase	NN	O	O
activity	NN	O	O
,	NN	O	O
although	NN	O	O
neither	NN	O	O
locus	NN	O	O
is	NN	O	O
an	NN	O	O
immunoglobulin	NN	O	B-protein
or	NN	O	O
a	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Two	NN	O	O
other	NN	O	O
T	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
and	NN	O	O
RPMI	NN	O	B-cell_line
8402	NN	O	I-cell_line
,	NN	O	O
have	NN	O	O
essentially	NN	O	O
identical	NN	O	O
deletions	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
growth-affecting	NN	O	B-DNA
genes	NN	O	I-DNA
other	NN	O	O
than	NN	O	O
immune	NN	O	B-protein
receptors	NN	O	I-protein
risk	NN	O	O
rearrangements	NN	O	O
.	NN	O	O

-DOCSTART-	O

Thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptors	NN	O	I-protein
form	NN	O	O
distinct	NN	O	O
nuclear	NN	O	B-protein
protein-dependent	NN	O	I-protein
and	NN	O	I-protein
independent	NN	O	I-protein
complexes	NN	O	I-protein
with	NN	O	O
a	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

We	NN	O	O
have	NN	O	O
examined	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
recombinant	NN	O	B-protein
thyroid	NN	O	I-protein
hormone	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
TRs	NN	O	B-protein
)	NN	O	O
to	NN	O	O
the	NN	O	O
palindromic	NN	O	B-DNA
thyroid	NN	O	I-DNA
hormone	NN	O	I-DNA
responsive	NN	O	I-DNA
element	NN	O	I-DNA
AGGTCATGACCT	NN	O	O
(	NN	O	O
TREp	NN	O	B-DNA
)	NN	O	O
using	NN	O	O
a	NN	O	O
gel	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	NN	O	O

Four	NN	O	O
specific	NN	O	O
protein-DNA	NN	O	B-protein
complexes	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
after	NN	O	O
incubation	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
(	NN	O	O
NE	NN	O	O
)	NN	O	O
from	NN	O	O
T3-responsive	NN	O	B-cell_line
pituitary	NN	O	I-cell_line
(	NN	O	I-cell_line
GH3	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
a	NN	O	O
TREp-containing	NN	O	B-DNA
DNA	NN	O	I-DNA
fragment	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
was	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
TREp	NN	O	B-DNA
binding	NN	O	O
of	NN	O	O
reticulocyte	NN	O	B-protein
lysate-synthesized	NN	O	I-protein
TRs	NN	O	I-protein
.	NN	O	O

TR	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
TR	NN	O	B-protein
beta	NN	O	I-protein
2	NN	O	I-protein
each	NN	O	O
formed	NN	O	O
a	NN	O	O
single	NN	O	O
major	NN	O	O
TR	NN	O	B-protein
:	NN	O	I-protein
TREp	NN	O	I-protein
complex	NN	O	I-protein
which	NN	O	O
comigrated	NN	O	O
with	NN	O	O
the	NN	O	O
least	NN	O	O
retarded	NN	O	O
complex	NN	O	O
formed	NN	O	O
by	NN	O	O
GH3	NN	O	O
NE	NN	O	O
,	NN	O	O
while	NN	O	O
TR	NN	O	B-protein
beta	NN	O	I-protein
1	NN	O	I-protein
formed	NN	O	O
multiple	NN	O	O
complexes	NN	O	O
suggesting	NN	O	O
that	NN	O	O
it	NN	O	O
can	NN	O	O
bind	NN	O	O
to	NN	O	O
TREp	NN	O	B-DNA
as	NN	O	O
an	NN	O	O
oligomer	NN	O	B-protein
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
coincubation	NN	O	O
of	NN	O	O
35S-TR	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
,	NN	O	O
GH3	NN	O	O
NE	NN	O	O
,	NN	O	O
and	NN	O	O
unlabeled	NN	O	O
TREp	NN	O	B-DNA
resulted	NN	O	O
in	NN	O	O
not	NN	O	O
only	NN	O	O
the	NN	O	O
35S-TR	NN	O	B-protein
:	NN	O	I-protein
TREp	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
but	NN	O	O
in	NN	O	O
two	NN	O	O
additional	NN	O	O
more	NN	O	O
greatly	NN	O	O
retarded	NN	O	O
complexes	NN	O	O
containing	NN	O	O
35S-TR	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
and	NN	O	O
comigrating	NN	O	O
with	NN	O	O
those	NN	O	O
formed	NN	O	O
by	NN	O	O
GH3	NN	O	O
extract	NN	O	O
alone	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
each	NN	O	O
of	NN	O	O
the	NN	O	O
TRs	NN	O	B-protein
with	NN	O	O
NE	NN	O	O
from	NN	O	O
COS-7	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
possess	NN	O	O
sufficient	NN	O	O
endogenous	NN	O	O
TRs	NN	O	B-protein
to	NN	O	O
mediate	NN	O	O
T3-responses	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
formation	NN	O	O
of	NN	O	O
a	NN	O	O
new	NN	O	O
,	NN	O	O
more	NN	O	O
greatly	NN	O	B-protein
shifted	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

A	NN	O	O
similar	NN	O	O
,	NN	O	O
heat	NN	O	O
labile	NN	O	O
activity	NN	O	O
which	NN	O	O
altered	NN	O	O
mobility	NN	O	O
of	NN	O	O
the	NN	O	O
TR	NN	O	B-protein
:	NN	O	I-protein
TRE	NN	O	I-protein
complex	NN	O	I-protein
was	NN	O	O
also	NN	O	O
present	NN	O	O
in	NN	O	O
NE	NN	O	O
from	NN	O	O
T3-unresponsive	NN	O	B-cell_line
JEG-3	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

At	NN	O	O
high	NN	O	O
concentration	NN	O	O
of	NN	O	O
NE	NN	O	O
,	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
TR	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
TREp	NN	O	B-DNA
was	NN	O	O
more	NN	O	O
greatly	NN	O	O
retarded	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
NE	NN	O	O
.	NN	O	O

Truncation	NN	O	O
of	NN	O	O
TR	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
at	NN	O	O
amino	NN	O	B-protein
acid	NN	O	I-protein
210	NN	O	I-protein
prevented	NN	O	O
additional	NN	O	O
complex	NN	O	O
formation	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
NE	NN	O	O
without	NN	O	O
affecting	NN	O	O
DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
carboxyl-terminus	NN	O	B-protein
of	NN	O	O
the	NN	O	O
TRs	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
interaction	NN	O	O
with	NN	O	O
nuclear	NN	O	B-protein
proteins	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

Cell-specific	NN	O	O
differences	NN	O	O
in	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
of	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
and	NN	O	I-DNA
beta	NN	O	I-DNA
interferon	NN	O	I-DNA
promoters	NN	O	I-DNA
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Three	NN	O	O
aspects	NN	O	O
of	NN	O	O
the	NN	O	O
involvement	NN	O	O
of	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
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
pathogenesis	NN	O	O
were	NN	O	O
examined	NN	O	O
.	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
mRNA	NN	O	I-RNA
production	NN	O	O
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
polymerase	NN	O	O
chain	NN	O	O
reaction	NN	O	O
amplification	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_line
U937	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
a	NN	O	O
chronically	NN	O	B-cell_line
HIV	NN	O	I-cell_line
infected	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
U9-IIIB	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

TNF-alpha	NN	O	B-RNA
RNA	NN	O	I-RNA
was	NN	O	O
undetectable	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
low	NN	O	O
constitutive	NN	O	O
level	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
U9-IIIB	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Paramyxovirus	NN	O	O
infection	NN	O	O
induced	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
steady-state	NN	O	O
level	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-RNA
RNA	NN	O	I-RNA
in	NN	O	O
U9-IIIB	NN	O	B-cell_line
cells	NN	O	I-cell_line
compared	NN	O	O
with	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
HIV-infected	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
cells	NN	O	I-cell_line
produced	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
than	NN	O	O
did	NN	O	O
normal	NN	O	O
cells	NN	O	O
after	NN	O	O
a	NN	O	O
secondary	NN	O	O
virus	NN	O	O
infection	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
TNF-alpha	NN	O	B-protein
on	NN	O	O
gene	NN	O	O
expression	NN	O	O
were	NN	O	O
examined	NN	O	O
by	NN	O	O
transient	NN	O	O
expression	NN	O	O
assays	NN	O	O
using	NN	O	O
reporter	NN	O	B-DNA
chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
plasmids	NN	O	I-DNA
linked	NN	O	O
to	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
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
(	NN	O	O
LTR	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
the	NN	O	O
beta	NN	O	B-DNA
interferon	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
U937	NN	O	B-cell_line
and	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
inducibility	NN	O	O
of	NN	O	O
the	NN	O	O
different	NN	O	O
hybrid	NN	O	B-DNA
promoters	NN	O	I-DNA
by	NN	O	O
TNF-alpha	NN	O	B-protein
or	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
varied	NN	O	O
in	NN	O	O
a	NN	O	O
cell	NN	O	O
type-	NN	O	O
and	NN	O	O
promoter	NN	O	O
context-specific	NN	O	O
manner	NN	O	O
;	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
gene	NN	O	O
activity	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-DNA
B-containing	NN	O	I-DNA
plasmids	NN	O	I-DNA
correlated	NN	O	O
directly	NN	O	O
with	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
.	NN	O	O

Although	NN	O	O
the	NN	O	O
intact	NN	O	O
beta	NN	O	B-DNA
interferon	NN	O	I-DNA
promoter	NN	O	I-DNA
was	NN	O	O
only	NN	O	O
weakly	NN	O	O
stimulated	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
or	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
multimers	NN	O	O
of	NN	O	O
the	NN	O	O
PRDII	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B-binding	NN	O	I-DNA
domain	NN	O	I-DNA
were	NN	O	O
inducible	NN	O	O
by	NN	O	O
both	NN	O	O
agents	NN	O	O
.	NN	O	O

TNF-alpha	NN	O	B-protein
was	NN	O	O
able	NN	O	O
to	NN	O	O
increase	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
TNF-alpha	NN	O	B-protein
did	NN	O	O
not	NN	O	O
induce	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
above	NN	O	O
a	NN	O	O
constitutive	NN	O	O
level	NN	O	O
of	NN	O	O
activity	NN	O	O
.	NN	O	O

This	NN	O	O
level	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-independent	NN	O	O
activity	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
sufficient	NN	O	O
for	NN	O	O
virus	NN	O	O
multiplication	NN	O	O
,	NN	O	O
since	NN	O	O
TNF-alpha	NN	O	B-protein
treatment	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
kinetics	NN	O	O
of	NN	O	O
de	NN	O	O
novo	NN	O	O
HIV	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
viral	NN	O	B-RNA
RNA	NN	O	I-RNA
production	NN	O	O
in	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
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
TNF-alpha	NN	O	B-protein
dramatically	NN	O	O
enhanced	NN	O	O
the	NN	O	O
spread	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
through	NN	O	O
the	NN	O	O
cell	NN	O	O
population	NN	O	O
and	NN	O	O
increased	NN	O	O
viral	NN	O	B-RNA
RNA	NN	O	I-RNA
synthesis	NN	O	O
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
HIV-1	NN	O	O
multiplication	NN	O	O
was	NN	O	O
stimulated	NN	O	O
by	NN	O	O
TNF-alpha	NN	O	B-protein
treatment	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
cis-linked	NN	O	B-DNA
regulatory	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
HLA	NN	O	B-DNA
DRA	NN	O	I-DNA
promoter	NN	O	I-DNA
by	NN	O	O
transcription	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Two	NN	O	O
consensus	NN	O	O
sequences	NN	O	O
,	NN	O	O
called	NN	O	O
X	NN	O	B-DNA
and	NN	O	I-DNA
Y	NN	O	I-DNA
boxes	NN	O	I-DNA
,	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
regulating	NN	O	O
expression	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
have	NN	O	O
been	NN	O	O
defined	NN	O	O
within	NN	O	O
the	NN	O	O
immediate	NN	O	O
upstream	NN	O	O
region	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
promoters	NN	O	I-DNA
.	NN	O	O

Unlike	NN	O	O
other	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
promoters	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
HLA-DR	NN	O	B-DNA
alpha	NN	O	I-DNA
(	NN	O	I-DNA
DRA	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
also	NN	O	O
contains	NN	O	O
one	NN	O	O
element	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
``	NN	O	B-DNA
octamer	NN	O	I-DNA
''	NN	O	I-DNA
motif	NN	O	I-DNA
of	NN	O	O
immunoglobulin	NN	O	B-DNA
variable	NN	O	I-DNA
region	NN	O	I-DNA
promoters	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
transcription	NN	O	O
.	NN	O	O

This	NN	O	O
``	NN	O	O
octamer	NN	O	B-DNA
``	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
DRA	NN	O	B-protein
appears	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
both	NN	O	O
the	NN	O	O
ubiquitous	NN	O	B-protein
(	NN	O	O
OTF-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
lymphoid-specific	NN	O	B-protein
(	NN	O	I-protein
OTF-2	NN	O	I-protein
)	NN	O	I-protein
``	NN	O	I-protein
octamer	NN	O	I-protein
''	NN	O	I-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
but	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
other	NN	O	O
distinct	NN	O	O
``	NN	O	B-protein
octamer	NN	O	I-protein
''	NN	O	I-protein
complex	NN	O	I-protein
was	NN	O	O
found	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
function	NN	O	O
of	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
developed	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
a	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
construct	NN	O	I-DNA
is	NN	O	O
transcribed	NN	O	O
more	NN	O	O
efficiently	NN	O	O
in	NN	O	O
extracts	NN	O	O
from	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
than	NN	O	O
in	NN	O	O
extracts	NN	O	O
from	NN	O	O
class	NN	O	B-cell_line
II-negative	NN	O	I-cell_line
HeLa	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

5	NN	O	O
'	NN	O	O
deletion	NN	O	O
constructs	NN	O	O
which	NN	O	O
lacked	NN	O	O
the	NN	O	O
Y	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
but	NN	O	O
retained	NN	O	O
the	NN	O	O
``	NN	O	B-DNA
octamer	NN	O	I-DNA
''	NN	O	I-DNA
motif	NN	O	I-DNA
and	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
were	NN	O	O
completely	NN	O	O
inactive	NN	O	O
,	NN	O	O
and	NN	O	O
internal	NN	O	O
deletion	NN	O	O
of	NN	O	O
the	NN	O	O
Y	NN	O	B-DNA
box	NN	O	I-DNA
reduced	NN	O	O
transcription	NN	O	O
by	NN	O	O
95	NN	O	O
%	NN	O	O
.	NN	O	O

Using	NN	O	O
supercoiled	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
linear	NN	O	O
templates	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
differences	NN	O	O
in	NN	O	O
transcription	NN	O	O
efficiencies	NN	O	O
from	NN	O	O
templates	NN	O	O
lacking	NN	O	O
or	NN	O	O
disrupting	NN	O	O
the	NN	O	O
X	NN	O	B-DNA
consensus	NN	O	I-DNA
element	NN	O	I-DNA
that	NN	O	O
reflect	NN	O	O
effects	NN	O	O
of	NN	O	O
random	NN	O	O
replacement	NN	O	O
of	NN	O	O
X	NN	O	B-DNA
box	NN	O	I-DNA
sequences	NN	O	I-DNA
in	NN	O	O
transient	NN	O	O
expression	NN	O	O
assays	NN	O	O
.	NN	O	O

Demonstration	NN	O	O
of	NN	O	O
the	NN	O	O
complete	NN	O	O
dependence	NN	O	O
on	NN	O	O
the	NN	O	O
Y	NN	O	B-DNA
box	NN	O	I-DNA
in	NN	O	O
this	NN	O	O
system	NN	O	O
suggests	NN	O	O
that	NN	O	O
,	NN	O	O
despite	NN	O	O
its	NN	O	O
demonstrated	NN	O	O
importance	NN	O	O
in	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
``	NN	O	I-DNA
octamer	NN	O	I-DNA
''	NN	O	I-DNA
does	NN	O	O
not	NN	O	O
utilize	NN	O	O
OTF-2	NN	O	B-protein
in	NN	O	O
a	NN	O	O
manner	NN	O	O
analogous	NN	O	O
to	NN	O	O
immunoglobulin	NN	O	B-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

Lipopolysaccharide	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
monocyte/macrophage-specific	NN	O	O
stimulator	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
.	NN	O	O

Lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
potently	NN	O	O
stimulates	NN	O	O
human	NN	O	B-DNA
immunodeficiency	NN	O	I-DNA
virus	NN	O	I-DNA
type	NN	O	I-DNA
1-long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	I-DNA
HIV-1-LTR	NN	O	I-DNA
)	NN	O	I-DNA
CAT	NN	O	I-DNA
constructs	NN	O	I-DNA
transfected	NN	O	O
into	NN	O	O
monocyte/macrophage-like	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
but	NN	O	O
not	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
.	NN	O	O

This	NN	O	O
effect	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	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
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
LPS	NN	O	O
induces	NN	O	O
a	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	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

LPS	NN	O	O
is	NN	O	O
also	NN	O	O
shown	NN	O	O
to	NN	O	O
dramatically	NN	O	O
increase	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
from	NN	O	O
a	NN	O	O
chronically	NN	O	B-cell_line
infected	NN	O	I-cell_line
monocyte/macrophage-like	NN	O	I-cell_line
cloned	NN	O	I-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
which	NN	O	O
produces	NN	O	O
very	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
at	NN	O	O
baseline	NN	O	O
.	NN	O	O

The	NN	O	O
stimulation	NN	O	O
of	NN	O	O
viral	NN	O	O
production	NN	O	O
from	NN	O	O
this	NN	O	O
cell	NN	O	O
line	NN	O	O
occurs	NN	O	O
only	NN	O	O
if	NN	O	O
these	NN	O	O
cells	NN	O	O
are	NN	O	O
treated	NN	O	O
with	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
before	NN	O	O
treatment	NN	O	O
with	NN	O	O
LPS	NN	O	O
.	NN	O	O

This	NN	O	O
stimulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-RNA
RNA	NN	O	I-RNA
and	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
.	NN	O	O

LPS	NN	O	O
is	NN	O	O
not	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
HIV-1	NN	O	O
production	NN	O	O
in	NN	O	O
a	NN	O	O
cloned	NN	O	O
T	NN	O	B-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
of	NN	O	O
LPS	NN	O	O
on	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
occurs	NN	O	O
at	NN	O	O
picogram	NN	O	O
per	NN	O	O
milliliter	NN	O	O
concentrations	NN	O	O
and	NN	O	O
may	NN	O	O
be	NN	O	O
clinically	NN	O	O
significant	NN	O	O
in	NN	O	O
understanding	NN	O	O
the	NN	O	O
variability	NN	O	O
of	NN	O	O
the	NN	O	O
natural	NN	O	O
history	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

Steroid	NN	O	O
dose	NN	O	O
sparing	NN	O	O
:	NN	O	O
pharmacodynamic	NN	O	O
responses	NN	O	O
to	NN	O	O
single	NN	O	O
versus	NN	O	O
divided	NN	O	O
doses	NN	O	O
of	NN	O	O
methylprednisolone	NN	O	O
in	NN	O	O
man	NN	O	O
.	NN	O	O

Inhibitory	NN	O	O
drug	NN	O	O
interactions	NN	O	O
affecting	NN	O	O
the	NN	O	O
metabolism	NN	O	O
of	NN	O	O
methylprednisolone	NN	O	O
(	NN	O	O
MP	NN	O	O
)	NN	O	O
may	NN	O	O
produce	NN	O	O
either	NN	O	O
steroid	NN	O	O
sparing	NN	O	O
or	NN	O	O
adverse	NN	O	O
effects	NN	O	O
partly	NN	O	O
by	NN	O	O
increasing	NN	O	O
the	NN	O	O
exposure	NN	O	O
time	NN	O	O
to	NN	O	O
the	NN	O	O
steroid	NN	O	O
.	NN	O	O

This	NN	O	O
phenomenon	NN	O	O
can	NN	O	O
be	NN	O	O
mimicked	NN	O	O
by	NN	O	O
administering	NN	O	O
MP	NN	O	O
in	NN	O	O
divided	NN	O	O
doses	NN	O	O
.	NN	O	O

Two	NN	O	O
types	NN	O	O
of	NN	O	O
responses	NN	O	O
were	NN	O	O
compared	NN	O	O
after	NN	O	O
a	NN	O	O
single	NN	O	O
MP	NN	O	O
dose	NN	O	O
(	NN	O	O
40	NN	O	O
mg	NN	O	O
bolus	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
divided	NN	O	O
regimen	NN	O	O
(	NN	O	O
20	NN	O	O
mg	NN	O	O
bolus	NN	O	O
and	NN	O	O
a	NN	O	O
5	NN	O	O
mg	NN	O	O
bolus	NN	O	O
8	NN	O	O
hours	NN	O	O
later	NN	O	O
)	NN	O	O
in	NN	O	O
six	NN	O	O
healthy	NN	O	O
male	NN	O	O
volunteers	NN	O	O
.	NN	O	O

The	NN	O	O
suppression	NN	O	O
of	NN	O	O
basophils	NN	O	B-cell_type
measured	NN	O	O
as	NN	O	O
whole	NN	O	O
blood	NN	O	O
histamine	NN	O	O
and	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
was	NN	O	O
assessed	NN	O	O
during	NN	O	O
32	NN	O	O
hours	NN	O	O
.	NN	O	O

The	NN	O	O
37.5	NN	O	O
%	NN	O	O
reduction	NN	O	O
in	NN	O	O
dose	NN	O	O
produced	NN	O	O
a	NN	O	O
23	NN	O	O
%	NN	O	O
overall	NN	O	O
decreased	NN	O	O
blood	NN	O	O
histamine	NN	O	O
response	NN	O	O
.	NN	O	O

A	NN	O	O
pharmacodynamic	NN	O	O
model	NN	O	O
for	NN	O	O
basophil	NN	O	O
cell	NN	O	O
distribution	NN	O	O
to	NN	O	O
and	NN	O	O
from	NN	O	O
an	NN	O	O
extravascular	NN	O	O
compartment	NN	O	O
describes	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
MP	NN	O	O
after	NN	O	O
both	NN	O	O
regimens	NN	O	O
.	NN	O	O

A	NN	O	O
slower	NN	O	O
initial	NN	O	O
decline	NN	O	O
in	NN	O	O
blood	NN	O	O
histamine	NN	O	O
after	NN	O	O
the	NN	O	O
divided	NN	O	O
regimen	NN	O	O
may	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
incomplete	NN	O	O
suppression	NN	O	O
of	NN	O	O
basophil	NN	O	B-cell_type
cell	NN	O	I-cell_type
return	NN	O	O
to	NN	O	O
blood	NN	O	O
.	NN	O	O

The	NN	O	O
50	NN	O	O
%	NN	O	O
inhibitory	NN	O	O
concentrations	NN	O	O
of	NN	O	O
MP	NN	O	O
of	NN	O	O
about	NN	O	O
5	NN	O	O
ng/ml	NN	O	O
were	NN	O	O
similar	NN	O	O
for	NN	O	O
both	NN	O	O
regimens	NN	O	O
.	NN	O	O

The	NN	O	O
decline	NN	O	O
and	NN	O	O
return	NN	O	O
of	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
were	NN	O	O
similar	NN	O	O
between	NN	O	O
MP	NN	O	O
treatments	NN	O	O
with	NN	O	O
suppression	NN	O	O
continuing	NN	O	O
for	NN	O	O
24	NN	O	O
hours	NN	O	O
.	NN	O	O

The	NN	O	O
50	NN	O	O
%	NN	O	O
inhibitory	NN	O	O
concentrations	NN	O	O
of	NN	O	O
MP	NN	O	O
values	NN	O	O
for	NN	O	O
adrenal	NN	O	O
suppression	NN	O	O
were	NN	O	O
about	NN	O	O
1	NN	O	O
ng/ml	NN	O	O
.	NN	O	O

Pharmacodynamic	NN	O	O
modeling	NN	O	O
is	NN	O	O
useful	NN	O	O
in	NN	O	O
quantitating	NN	O	O
corticosteroid	NN	O	O
responses	NN	O	O
and	NN	O	O
generally	NN	O	O
predicted	NN	O	O
the	NN	O	O
``	NN	O	O
dose-sparing	NN	O	O
''	NN	O	O
effects	NN	O	O
that	NN	O	O
were	NN	O	O
achieved	NN	O	O
by	NN	O	O
prolonging	NN	O	O
MP	NN	O	O
plasma	NN	O	O
concentrations	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
supports	NN	O	O
previous	NN	O	O
clinical	NN	O	O
observations	NN	O	O
that	NN	O	O
patients	NN	O	O
may	NN	O	O
require	NN	O	O
morning	NN	O	O
through	NN	O	O
evening	NN	O	O
exposure	NN	O	O
to	NN	O	O
MP	NN	O	O
to	NN	O	O
optimize	NN	O	O
efficacy	NN	O	O
while	NN	O	O
adrenal	NN	O	O
suppression	NN	O	O
is	NN	O	O
being	NN	O	O
minimized	NN	O	O
.	NN	O	O

-DOCSTART-	O

1	NN	O	O
,	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D2	NN	O	O
production	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
alveolar	NN	O	B-cell_type
macrophages	NN	O	I-cell_type
recovered	NN	O	O
by	NN	O	O
lavage	NN	O	O
from	NN	O	O
normocalcemic	NN	O	O
patients	NN	O	O
with	NN	O	O
tuberculosis	NN	O	O
.	NN	O	O

To	NN	O	O
compare	NN	O	O
extra-renal	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
production	NN	O	O
in	NN	O	O
different	NN	O	O
types	NN	O	O
of	NN	O	O
granulomatous	NN	O	O
disease	NN	O	O
,	NN	O	O
and	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
cell	NN	O	O
types	NN	O	O
responsible	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
evaluated	NN	O	O
the	NN	O	O
conversion	NN	O	O
of	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
D3	NN	O	O
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
by	NN	O	O
uncultured	NN	O	B-cell_type
cells	NN	O	I-cell_type
recovered	NN	O	O
by	NN	O	O
bronchoalveolar	NN	O	O
lavage	NN	O	O
and	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
normocalcemic	NN	O	O
patients	NN	O	O
with	NN	O	O
sarcoidosis	NN	O	O
and	NN	O	O
tuberculosis	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
was	NN	O	O
produced	NN	O	O
both	NN	O	O
by	NN	O	O
lavage	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
12/12	NN	O	O
tuberculosis	NN	O	O
patients	NN	O	O
,	NN	O	O
2/6	NN	O	O
sarcoidosis	NN	O	O
patients	NN	O	O
)	NN	O	O
and	NN	O	O
blood	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
3/5	NN	O	O
tuberculosis	NN	O	O
patients	NN	O	O
,	NN	O	O
0/3	NN	O	O
sarcoidosis	NN	O	O
patients	NN	O	O
)	NN	O	O
from	NN	O	O
patients	NN	O	O
but	NN	O	O
not	NN	O	O
controls	NN	O	O
,	NN	O	O
but	NN	O	O
significantly	NN	O	O
greater	NN	O	O
amounts	NN	O	O
were	NN	O	O
produced	NN	O	O
by	NN	O	O
lavage	NN	O	O
cells	NN	O	O
from	NN	O	O
tuberculosis	NN	O	O
patients	NN	O	O
than	NN	O	O
those	NN	O	O
of	NN	O	O
sarcoidosis	NN	O	O
patients	NN	O	O
(	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

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
production	NN	O	O
by	NN	O	O
lavage	NN	O	B-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
tuberculosis	NN	O	O
patients	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
CD8+	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
present	NN	O	O
but	NN	O	O
not	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
appeared	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
source	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
production	NN	O	O
,	NN	O	O
since	NN	O	O
purified	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
from	NN	O	O
all	NN	O	O
patients	NN	O	O
with	NN	O	O
tuberculosis	NN	O	O
produced	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
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
production	NN	O	O
by	NN	O	O
these	NN	O	O
cells	NN	O	O
correlated	NN	O	O
closely	NN	O	O
with	NN	O	O
that	NN	O	O
produced	NN	O	O
by	NN	O	O
unseparated	NN	O	O
lavage	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Because	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
improve	NN	O	O
the	NN	O	O
capacity	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
to	NN	O	O
kill	NN	O	O
mycobacteria	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
support	NN	O	O
the	NN	O	O
conclusion	NN	O	O
that	NN	O	O
macrophage-lymphocyte	NN	O	O
interactions	NN	O	O
,	NN	O	O
mediated	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	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
may	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
component	NN	O	O
of	NN	O	O
a	NN	O	O
successful	NN	O	O
antituberculous	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Megakaryocytic	NN	O	B-cell_line
and	NN	O	I-cell_line
erythrocytic	NN	O	I-cell_line
lineages	NN	O	I-cell_line
share	NN	O	O
specific	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Erythroid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
contain	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
NF-E1	NN	O	B-protein
(	NN	O	O
also	NN	O	O
called	NN	O	O
GF-1	NN	O	B-protein
and	NN	O	O
Eryf-1	NN	O	B-protein
;	NN	O	O
refs	NN	O	O
1-3	NN	O	O
respectively	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
principal	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
the	NN	O	O
erythrocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

NF-E1	NN	O	B-protein
expression	NN	O	O
seems	NN	O	O
to	NN	O	O
be	NN	O	O
restricted	NN	O	O
to	NN	O	O
the	NN	O	O
erythrocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

A	NN	O	O
closely	NN	O	O
related	NN	O	O
(	NN	O	O
if	NN	O	O
not	NN	O	O
identical	NN	O	O
)	NN	O	O
protein	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
both	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
megakaryocytic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
purified	NN	O	B-cell_line
human	NN	O	I-cell_line
megakaryocytes	NN	O	I-cell_line
;	NN	O	O
it	NN	O	O
binds	NN	O	O
to	NN	O	O
promoter	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
two	NN	O	O
megakaryocytic-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
binding	NN	O	O
sites	NN	O	O
and	NN	O	O
partial	NN	O	O
proteolysis	NN	O	O
profile	NN	O	O
of	NN	O	O
this	NN	O	O
protein	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
erythroid	NN	O	B-protein
protein	NN	O	I-protein
;	NN	O	O
also	NN	O	O
,	NN	O	O
NF-E1	NN	O	B-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
is	NN	O	O
the	NN	O	O
same	NN	O	O
size	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
megakaryocytic	NN	O	B-cell_line
and	NN	O	I-cell_line
erythroid	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
point	NN	O	O
mutations	NN	O	O
that	NN	O	O
abolish	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-E1	NN	O	B-protein
result	NN	O	O
in	NN	O	O
a	NN	O	O
70	NN	O	O
%	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
megakaryocytic-specific	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
also	NN	O	O
find	NN	O	O
that	NN	O	O
NF-E2	NN	O	B-protein
,	NN	O	O
another	NN	O	O
trans-acting	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	O
the	NN	O	O
erythrocytic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
megakaryocytes	NN	O	B-cell_line
.	NN	O	O

Transcriptional	NN	O	O
effects	NN	O	O
in	NN	O	O
both	NN	O	O
lineages	NN	O	O
might	NN	O	O
then	NN	O	O
be	NN	O	O
mediated	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
the	NN	O	O
same	NN	O	O
specific	NN	O	O
trans-acting	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
data	NN	O	O
strengthen	NN	O	O
the	NN	O	O
idea	NN	O	O
of	NN	O	O
a	NN	O	O
close	NN	O	O
association	NN	O	O
between	NN	O	O
the	NN	O	O
erythrocytic	NN	O	B-cell_line
and	NN	O	I-cell_line
the	NN	O	I-cell_line
megakaryocytic	NN	O	I-cell_line
lineages	NN	O	I-cell_line
and	NN	O	O
could	NN	O	O
also	NN	O	O
explain	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
markers	NN	O	O
specific	NN	O	O
to	NN	O	O
the	NN	O	O
erythrocytic	NN	O	B-cell_type
and	NN	O	I-cell_type
megakaryocytic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
in	NN	O	O
most	NN	O	O
erythroblastic	NN	O	B-cell_line
and	NN	O	I-cell_line
megakaryoblastic	NN	O	I-cell_line
permanent	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
c-myc	NN	O	B-DNA
expression	NN	O	O
by	NN	O	O
protein	NN	O	O
synthesis-dependent	NN	O	O
and	NN	O	O
-independent	NN	O	O
pathways	NN	O	O
in	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphoblastic	NN	O	I-cell_line
tumor	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
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphoblastic	NN	O	I-cell_line
tumor	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
Molt4	NN	O	B-RNA
c-myc	NN	O	I-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
is	NN	O	O
down-regulated	NN	O	O
after	NN	O	O
exposure	NN	O	O
to	NN	O	O
dimethyl	NN	O	O
sulfoxide	NN	O	O
,	NN	O	O
to	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
,	NN	O	O
or	NN	O	O
to	NN	O	O
the	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
A23187	NN	O	O
,	NN	O	O
which	NN	O	O
raises	NN	O	O
the	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
concentration	NN	O	O
.	NN	O	O

A	NN	O	O
block	NN	O	O
to	NN	O	O
RNA	NN	O	O
elongation	NN	O	O
is	NN	O	O
largely	NN	O	O
responsible	NN	O	O
for	NN	O	O
decreased	NN	O	O
c-myc	NN	O	B-DNA
transcription	NN	O	O
.	NN	O	O

Although	NN	O	O
negative	NN	O	O
regulation	NN	O	O
by	NN	O	O
dimethyl	NN	O	O
sulfoxide	NN	O	O
takes	NN	O	O
place	NN	O	O
even	NN	O	O
when	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
is	NN	O	O
inhibited	NN	O	O
by	NN	O	O
cycloheximide	NN	O	O
,	NN	O	O
the	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
effect	NN	O	O
is	NN	O	O
blocked	NN	O	O
to	NN	O	O
some	NN	O	O
extent	NN	O	O
only	NN	O	O
by	NN	O	O
cycloheximide	NN	O	O
.	NN	O	O

The	NN	O	O
calcium	NN	O	O
ionophore-induced	NN	O	O
c-myc	NN	O	B-DNA
suppression	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
strictly	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

Therefore	NN	O	O
,	NN	O	O
two	NN	O	O
different	NN	O	O
negative	NN	O	O
regulatory	NN	O	O
pathways	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
c-myc	NN	O	B-DNA
regulation	NN	O	O
:	NN	O	O
one	NN	O	O
which	NN	O	O
is	NN	O	O
independent	NN	O	O
and	NN	O	O
one	NN	O	O
which	NN	O	O
depends	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

The	NN	O	O
latter	NN	O	O
one	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
a	NN	O	O
rapidly	NN	O	O
calcium-dependent	NN	O	B-protein
induced	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Oestrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
analysis	NN	O	O
in	NN	O	O
B-cell	NN	O	O
chronic	NN	O	O
lymphocytic	NN	O	O
leukemia	NN	O	O
:	NN	O	O
correlation	NN	O	O
of	NN	O	O
biochemical	NN	O	O
and	NN	O	O
immunocytochemical	NN	O	O
methods	NN	O	O
.	NN	O	O

Oestrogen	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
ER	NN	O	B-protein
)	NN	O	O
are	NN	O	O
present	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
and	NN	O	O
have	NN	O	O
been	NN	O	O
considered	NN	O	O
a	NN	O	O
physiological	NN	O	O
marker	NN	O	O
of	NN	O	O
growth	NN	O	O
rate	NN	O	O
or	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Tamoxifen	NN	O	O
,	NN	O	O
an	NN	O	O
oestrogen	NN	O	O
antagonist	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
given	NN	O	O
in	NN	O	O
some	NN	O	O
patients	NN	O	O
with	NN	O	O
CLL	NN	O	O
and	NN	O	O
Hodgkin	NN	O	O
's	NN	O	O
disease	NN	O	O
,	NN	O	O
with	NN	O	O
dramatic	NN	O	O
response	NN	O	O
in	NN	O	O
single	NN	O	O
cases	NN	O	O
.	NN	O	O

Until	NN	O	O
now	NN	O	O
,	NN	O	O
ER	NN	O	B-protein
status	NN	O	O
has	NN	O	O
been	NN	O	O
assessed	NN	O	O
using	NN	O	O
a	NN	O	O
steroid	NN	O	O
binding	NN	O	O
assay	NN	O	O
(	NN	O	O
SBA	NN	O	O
)	NN	O	O
which	NN	O	O
has	NN	O	O
many	NN	O	O
inherent	NN	O	O
problems	NN	O	O
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
monoclonal	NN	O	O
antibodies	NN	O	O
directed	NN	O	O
against	NN	O	O
ER	NN	O	B-protein
has	NN	O	O
been	NN	O	O
applied	NN	O	O
to	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
breast	NN	O	O
carcinomas	NN	O	O
and	NN	O	O
results	NN	O	O
obtained	NN	O	O
show	NN	O	O
good	NN	O	O
correlation	NN	O	O
with	NN	O	O
the	NN	O	O
quantitative	NN	O	O
SBA	NN	O	O
.	NN	O	O

We	NN	O	O
studied	NN	O	O
49	NN	O	O
cases	NN	O	O
of	NN	O	O
B-cell	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
using	NN	O	O
immunostaining	NN	O	O
of	NN	O	O
cytospin	NN	O	O
preparations	NN	O	O
.	NN	O	O

In	NN	O	O
30	NN	O	O
of	NN	O	O
these	NN	O	O
cases	NN	O	O
ER	NN	O	B-protein
enzyme	NN	O	O
immunoassay	NN	O	O
(	NN	O	O
ER	NN	O	B-protein
-EIA	NN	O	O
)	NN	O	O
was	NN	O	O
also	NN	O	O
performed	NN	O	O
.	NN	O	O

Cultured	NN	O	B-cell_line
MCF-7	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
derived	NN	O	O
from	NN	O	O
a	NN	O	O
pleural	NN	O	O
effusion	NN	O	O
of	NN	O	O
a	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patient	NN	O	O
,	NN	O	O
known	NN	O	O
to	NN	O	O
contain	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
were	NN	O	O
used	NN	O	O
as	NN	O	O
a	NN	O	O
positive	NN	O	O
control	NN	O	O
(	NN	O	O
40-48	NN	O	O
%	NN	O	O
ER	NN	O	B-cell_type
positive	NN	O	I-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
immunocytochemistry	NN	O	O
;	NN	O	O
200	NN	O	O
fmol/mg	NN	O	O
protein	NN	O	O
by	NN	O	O
EIA	NN	O	O
)	NN	O	O
.	NN	O	O

All	NN	O	O
of	NN	O	O
the	NN	O	O
CLL	NN	O	O
cases	NN	O	O
except	NN	O	O
two	NN	O	O
(	NN	O	O
96	NN	O	O
%	NN	O	O
)	NN	O	O
were	NN	O	O
negative	NN	O	O
for	NN	O	O
ER	NN	O	B-protein
(	NN	O	O
less	NN	O	O
than	NN	O	O
1	NN	O	O
%	NN	O	O
staining	NN	O	O
;	NN	O	O
less	NN	O	O
than	NN	O	O
4	NN	O	O
fmol/mg	NN	O	O
protein	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
positive	NN	O	O
cases	NN	O	O
expressed	NN	O	O
granular	NN	O	O
ER	NN	O	B-protein
staining	NN	O	O
over	NN	O	O
the	NN	O	O
nucleus	NN	O	O
(	NN	O	O
9.2	NN	O	O
and	NN	O	O
12.1	NN	O	O
%	NN	O	O
positive	NN	O	O
cells	NN	O	O
)	NN	O	O
and	NN	O	O
were	NN	O	O
positive	NN	O	O
by	NN	O	O
EIA	NN	O	O
and	NN	O	O
SBA	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
i	NN	O	O
)	NN	O	O
patients	NN	O	O
with	NN	O	O
CLL	NN	O	O
rarely	NN	O	O
express	NN	O	O
ER	NN	O	B-protein
and	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
immunocytochemical	NN	O	O
staining	NN	O	O
of	NN	O	O
cytospin	NN	O	O
preparations	NN	O	O
is	NN	O	O
a	NN	O	O
valid	NN	O	O
technique	NN	O	O
for	NN	O	O
the	NN	O	O
measurement	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
of	NN	O	O
interest	NN	O	O
that	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
positive	NN	O	O
cases	NN	O	O
was	NN	O	O
diagnosed	NN	O	O
as	NN	O	O
CLL	NN	O	O
with	NN	O	O
Richter	NN	O	O
's	NN	O	O
transformation	NN	O	O
confirming	NN	O	O
earlier	NN	O	O
findings	NN	O	O
.	NN	O	O

-DOCSTART-	O

Type-II	NN	O	B-protein
estrogen	NN	O	I-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
in	NN	O	O
a	NN	O	O
lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
growth-inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
estrogen	NN	O	O
,	NN	O	O
anti-estrogen	NN	O	O
and	NN	O	O
bioflavonoids	NN	O	O
.	NN	O	O

Type-II	NN	O	B-protein
estrogen-binding	NN	O	I-protein
sites	NN	O	I-protein
(	NN	O	O
type-II	NN	O	B-protein
EBS	NN	O	I-protein
)	NN	O	O
have	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
in	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
IM-9	NN	O	B-cell_line
using	NN	O	O
a	NN	O	O
whole-cell	NN	O	O
assay	NN	O	O
with	NN	O	O
(	NN	O	O
6	NN	O	O
,	NN	O	O
7-3H	NN	O	O
)	NN	O	O
estradiol	NN	O	O
(	NN	O	O
3H-E2	NN	O	O
)	NN	O	O
as	NN	O	O
tracer	NN	O	O
.	NN	O	O

Competition	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
anti-estrogen	NN	O	O
tamoxifen	NN	O	O
and	NN	O	O
the	NN	O	O
flavonoids	NN	O	O
quercetin	NN	O	O
and	NN	O	O
rutin	NN	O	O
competed	NN	O	O
for	NN	O	O
(	NN	O	O
3H	NN	O	O
)	NN	O	O
-E2	NN	O	O
binding	NN	O	O
to	NN	O	O
type-II	NN	O	B-protein
EBS	NN	O	I-protein
.	NN	O	O

Growth	NN	O	O
experiments	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
diethylstilbestrol	NN	O	O
(	NN	O	O
DES	NN	O	O
)	NN	O	O
tamoxifen	NN	O	O
(	NN	O	O
TAM	NN	O	O
)	NN	O	O
,	NN	O	O
quercetin	NN	O	O
and	NN	O	O
rutin	NN	O	O
exerted	NN	O	O
a	NN	O	O
reversible	NN	O	O
dose-dependent	NN	O	O
inhibition	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
in	NN	O	O
the	NN	O	O
range	NN	O	O
of	NN	O	O
concentrations	NN	O	O
between	NN	O	O
10	NN	O	O
nM	NN	O	O
and	NN	O	O
10	NN	O	O
microM	NN	O	O
.	NN	O	O

The	NN	O	O
relative	NN	O	O
binding	NN	O	O
affinity	NN	O	O
of	NN	O	O
quercetin	NN	O	O
,	NN	O	O
rutin	NN	O	O
,	NN	O	O
DES	NN	O	O
and	NN	O	O
TAM	NN	O	O
for	NN	O	O
type-II	NN	O	B-protein
EBS	NN	O	I-protein
correlated	NN	O	O
well	NN	O	O
with	NN	O	O
their	NN	O	O
potency	NN	O	O
as	NN	O	O
cell	NN	O	O
growth	NN	O	O
inhibitors	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
hesperidin	NN	O	O
,	NN	O	O
a	NN	O	O
flavonoid	NN	O	O
which	NN	O	O
does	NN	O	O
not	NN	O	O
bind	NN	O	O
to	NN	O	O
type-II	NN	O	B-protein
EBS	NN	O	I-protein
,	NN	O	O
was	NN	O	O
ineffective	NN	O	O
in	NN	O	O
inhibiting	NN	O	O
cell	NN	O	O
growth	NN	O	O
.	NN	O	O

Cell-cycle	NN	O	O
analysis	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
growth-inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
DES	NN	O	O
,	NN	O	O
TAM	NN	O	O
or	NN	O	O
quercetin	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
blocking	NN	O	O
effect	NN	O	O
in	NN	O	O
the	NN	O	O
G0-G1	NN	O	O
phases	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
high	NN	O	O
estrogen	NN	O	O
and	NN	O	O
anti-estrogen	NN	O	O
concentrations	NN	O	O
and	NN	O	O
flavonoids	NN	O	O
may	NN	O	O
regulate	NN	O	O
IM-9	NN	O	B-cell_line
cell	NN	O	O
growth	NN	O	O
through	NN	O	O
a	NN	O	O
common	NN	O	O
mechanism	NN	O	O
involving	NN	O	O
a	NN	O	O
binding	NN	O	O
interaction	NN	O	O
with	NN	O	O
type-II	NN	O	B-protein
EBS	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Glucocorticoid	NN	O	B-protein
receptors	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
of	NN	O	O
patients	NN	O	O
with	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
]	NN	O	O

Quantitation	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
and	NN	O	O
the	NN	O	O
study	NN	O	O
of	NN	O	O
their	NN	O	O
affinity	NN	O	O
for	NN	O	O
glucocorticosteroids	NN	O	O
(	NN	O	O
GCS	NN	O	O
)	NN	O	O
were	NN	O	O
made	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
of	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
(	NN	O	O
BA	NN	O	O
)	NN	O	O
patients	NN	O	O
in	NN	O	O
consideration	NN	O	O
of	NN	O	O
GCR	NN	O	B-protein
treatment	NN	O	O
and	NN	O	O
serum	NN	O	O
levels	NN	O	O
of	NN	O	O
endogenous	NN	O	O
cortisol	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
stated	NN	O	O
that	NN	O	O
GCR	NN	O	B-protein
of	NN	O	O
healthy	NN	O	O
controls	NN	O	O
and	NN	O	O
GCS-untreated	NN	O	O
patients	NN	O	O
outnumbered	NN	O	O
those	NN	O	O
of	NN	O	O
cortisol-dependent	NN	O	O
BA	NN	O	O
patients	NN	O	O
on	NN	O	O
hormone	NN	O	O
therapy	NN	O	O
.	NN	O	O

Following	NN	O	O
discontinuation	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
drugs	NN	O	O
GCR	NN	O	B-protein
count	NN	O	O
in	NN	O	O
cortisol-dependent	NN	O	O
BA	NN	O	O
tends	NN	O	O
to	NN	O	O
rise	NN	O	O
.	NN	O	O

Endogenous	NN	O	O
cortisol	NN	O	O
has	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
GCR	NN	O	B-protein
level	NN	O	O
estimated	NN	O	O
by	NN	O	O
3H-triamcinolone	NN	O	O
acetonide	NN	O	O
.	NN	O	O

-DOCSTART-	O

Two	NN	O	O
glucocorticoid	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
on	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

Glucocorticoids	NN	O	O
are	NN	O	O
known	NN	O	O
to	NN	O	O
have	NN	O	O
a	NN	O	O
lytic	NN	O	O
effect	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
via	NN	O	O
interactions	NN	O	O
with	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

Cortisol	NN	O	O
and	NN	O	O
various	NN	O	O
synthetic	NN	O	O
glucocorticoids	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
with	NN	O	O
one-site	NN	O	O
kinetics	NN	O	O
.	NN	O	O

Cortivazol	NN	O	O
(	NN	O	O
CVZ	NN	O	O
)	NN	O	O
is	NN	O	O
a	NN	O	O
unique	NN	O	O
,	NN	O	O
high	NN	O	O
potency	NN	O	O
synthetic	NN	O	O
glucocorticoid	NN	O	O
,	NN	O	O
which	NN	O	O
has	NN	O	O
a	NN	O	O
phenylpyrazol	NN	O	O
fused	NN	O	O
to	NN	O	O
the	NN	O	O
A-ring	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	O
nucleus	NN	O	O
and	NN	O	O
displays	NN	O	O
binding	NN	O	O
consistent	NN	O	O
with	NN	O	O
two	NN	O	O
or	NN	O	O
more	NN	O	O
sites	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
from	NN	O	O
CEM	NN	O	B-cell_line
C7	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
acute	NN	O	I-cell_line
lymphoblastic	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
)	NN	O	O
.	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
the	NN	O	O
lower	NN	O	O
affinity	NN	O	O
class	NN	O	O
of	NN	O	O
sites	NN	O	O
are	NN	O	O
similar	NN	O	O
in	NN	O	O
affinity	NN	O	O
and	NN	O	O
site	NN	O	O
molarity	NN	O	O
to	NN	O	O
those	NN	O	O
recognized	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

The	NN	O	O
higher	NN	O	O
affinity	NN	O	O
sites	NN	O	O
bind	NN	O	O
CVZ	NN	O	O
with	NN	O	O
20-	NN	O	O
to	NN	O	O
50-fold	NN	O	O
greater	NN	O	O
affinity	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
CVZ	NN	O	O
's	NN	O	O
enhanced	NN	O	O
biological	NN	O	O
effects	NN	O	O
.	NN	O	O

In	NN	O	O
mutant	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
resistant	NN	O	O
to	NN	O	O
the	NN	O	O
lytic	NN	O	O
effects	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
,	NN	O	O
CVZ	NN	O	O
both	NN	O	O
lyses	NN	O	O
the	NN	O	O
cells	NN	O	O
and	NN	O	O
recognizes	NN	O	O
a	NN	O	O
single	NN	O	O
class	NN	O	O
of	NN	O	O
sites	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
high	NN	O	O
affinity	NN	O	O
site	NN	O	O
in	NN	O	O
CEM	NN	O	B-cell_line
C7	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
have	NN	O	O
carried	NN	O	O
out	NN	O	O
experiments	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
higher	NN	O	B-protein
affinity	NN	O	I-protein
CVZ	NN	O	I-protein
binding	NN	O	I-protein
site	NN	O	I-protein
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
:	NN	O	O
1	NN	O	O
)	NN	O	O
CVZ	NN	O	O
has	NN	O	O
more	NN	O	O
than	NN	O	O
one	NN	O	O
binding	NN	O	O
site	NN	O	O
in	NN	O	O
a	NN	O	O
second	NN	O	O
,	NN	O	O
independent	NN	O	O
,	NN	O	O
B-cell	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
IM-9	NN	O	B-cell_line
;	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
antiglucocorticoid	NN	O	O
RU	NN	O	O
38486	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
block	NN	O	O
both	NN	O	O
CVZ	NN	O	O
's	NN	O	O
higher	NN	O	O
and	NN	O	O
lower	NN	O	O
affinity	NN	O	O
sites	NN	O	O
;	NN	O	O
3	NN	O	O
)	NN	O	O
all	NN	O	O
of	NN	O	O
CVZ	NN	O	B-protein
's	NN	O	I-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
are	NN	O	O
on	NN	O	O
a	NN	O	O
protein	NN	O	O
immunologically	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
the	NN	O	O
human	NN	O	O
GR	NN	O	B-protein
;	NN	O	O
and	NN	O	O
4	NN	O	O
)	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
clones	NN	O	O
of	NN	O	O
CVZ-resistant	NN	O	O
cells	NN	O	O
have	NN	O	O
lost	NN	O	O
all	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
CVZ	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
CVZ	NN	O	O
is	NN	O	O
recognizing	NN	O	O
two	NN	O	O
glucocorticoid	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
on	NN	O	O
the	NN	O	O
human	NN	O	O
GR	NN	O	B-protein
or	NN	O	O
a	NN	O	O
protein	NN	O	O
very	NN	O	O
similar	NN	O	O
to	NN	O	O
it	NN	O	O
.	NN	O	O

-DOCSTART-	O

Retroviral	NN	O	O
mediated	NN	O	O
transfer	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
exogenous	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
primary	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
assaying	NN	O	O
for	NN	O	O
a	NN	O	O
viral	NN	O	O
transactivator	NN	O	B-protein
activity	NN	O	O
in	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
.	NN	O	O

In	NN	O	O
this	NN	O	O
report	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
recombinant	NN	O	O
retroviruses	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
an	NN	O	O
exogenous	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
primary	NN	O	O
cells	NN	O	O
obtained	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
lymphoproliferative	NN	O	O
disorders	NN	O	O
.	NN	O	O

The	NN	O	O
infection	NN	O	O
of	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
cultured	NN	O	O
and	NN	O	O
primary	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
a	NN	O	O
recombinant	NN	O	O
retrovirus	NN	O	O
containing	NN	O	O
a	NN	O	O
histone	NN	O	B-DNA
promoter-driven	NN	O	I-DNA
beta-galactosidase	NN	O	I-DNA
gene	NN	O	I-DNA
is	NN	O	O
shown	NN	O	O
to	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
beta-galactosidase	NN	O	B-protein
in	NN	O	O
50	NN	O	O
%	NN	O	O
to	NN	O	O
100	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

A	NN	O	O
similar	NN	O	O
infection	NN	O	O
with	NN	O	O
a	NN	O	O
recombinant	NN	O	O
retrovirus	NN	O	O
containing	NN	O	O
the	NN	O	O
beta-galactosidase	NN	O	B-DNA
gene	NN	O	I-DNA
with	NN	O	O
an	NN	O	O
adenovirus	NN	O	B-DNA
E2	NN	O	I-DNA
promoter	NN	O	I-DNA
,	NN	O	O
results	NN	O	O
in	NN	O	O
beta-galactosidase	NN	O	B-protein
activity	NN	O	O
in	NN	O	O
a	NN	O	O
limited	NN	O	O
number	NN	O	O
of	NN	O	O
cultured	NN	O	B-cell_line
and	NN	O	O
primary	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Since	NN	O	O
the	NN	O	O
adenovirus	NN	O	B-DNA
E2	NN	O	I-DNA
promoter	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
well	NN	O	O
characterized	NN	O	O
and	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
transactivators	NN	O	B-protein
encoded	NN	O	O
by	NN	O	O
many	NN	O	O
viruses	NN	O	O
,	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
this	NN	O	O
promoter	NN	O	O
in	NN	O	O
specific	NN	O	O
cell	NN	O	O
types	NN	O	O
is	NN	O	O
discussed	NN	O	O
in	NN	O	O
reference	NN	O	O
to	NN	O	O
both	NN	O	O
the	NN	O	O
biology	NN	O	O
of	NN	O	O
the	NN	O	O
cell	NN	O	O
and	NN	O	O
the	NN	O	O
possible	NN	O	O
presence	NN	O	O
of	NN	O	O
as	NN	O	O
yet	NN	O	O
undetected	NN	O	O
viral	NN	O	B-protein
gene	NN	O	I-protein
products	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
The	NN	O	O
effect	NN	O	O
of	NN	O	O
24	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
dioxyvit	NN	O	O
)	NN	O	O
on	NN	O	O
Ca	NN	O	O
metabolism	NN	O	O
and	NN	O	O
immune	NN	O	O
status	NN	O	O
during	NN	O	O
chronic	NN	O	O
kidney	NN	O	O
failure	NN	O	O
]	NN	O	O

Active	NN	O	O
metabolite	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
,	NN	O	O
24R	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
dioxyvit	NN	O	O
)	NN	O	O
was	NN	O	O
used	NN	O	O
at	NN	O	O
a	NN	O	O
daily	NN	O	O
dose	NN	O	O
of	NN	O	O
100	NN	O	O
micrograms	NN	O	O
in	NN	O	O
treatment	NN	O	O
of	NN	O	O
children	NN	O	O
affected	NN	O	O
with	NN	O	O
tubulointerstitial	NN	O	O
disease	NN	O	O
of	NN	O	O
kidney	NN	O	O
and	NN	O	O
with	NN	O	O
chronic	NN	O	O
glomerulonephritis	NN	O	O
under	NN	O	O
conditions	NN	O	O
of	NN	O	O
kidney	NN	O	O
insufficiency	NN	O	O
.	NN	O	O

The	NN	O	O
drug	NN	O	O
exhibited	NN	O	O
distinct	NN	O	O
normalizing	NN	O	O
effect	NN	O	O
on	NN	O	O
patterns	NN	O	O
of	NN	O	O
calcium	NN	O	O
metabolism	NN	O	O
:	NN	O	O
increase	NN	O	O
of	NN	O	O
total	NN	O	O
and	NN	O	O
ionized	NN	O	O
Ca2+	NN	O	O
and	NN	O	O
of	NN	O	O
25-OHD	NN	O	O
,	NN	O	O
decrease	NN	O	O
in	NN	O	O
concentration	NN	O	O
of	NN	O	O
parath	NN	O	O
hormone	NN	O	O
and	NN	O	O
osteocalcine	NN	O	O
in	NN	O	O
blood	NN	O	O
serum	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
on	NN	O	O
immunological	NN	O	O
parameters	NN	O	O
:	NN	O	O
restoration	NN	O	O
of	NN	O	O
decreased	NN	O	O
content	NN	O	O
of	NN	O	O
T-	NN	O	B-cell_type
and	NN	O	I-cell_type
0-lymphocytes	NN	O	I-cell_type
.	NN	O	O

Concentration	NN	O	O
of	NN	O	O
receptors	NN	O	O
of	NN	O	O
hormonal	NN	O	O
form	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
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
minimal	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
under	NN	O	O
conditions	NN	O	O
of	NN	O	O
chronic	NN	O	O
kidney	NN	O	O
insufficiency	NN	O	O
,	NN	O	O
while	NN	O	O
their	NN	O	O
expression	NN	O	O
,	NN	O	O
after	NN	O	O
the	NN	O	O
dioxyvit	NN	O	O
action	NN	O	O
,	NN	O	O
was	NN	O	O
detected	NN	O	O
only	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
glomerulonephritis	NN	O	O
.	NN	O	O

Specific	NN	O	O
calcitropic	NN	O	O
effect	NN	O	O
of	NN	O	O
dioxyvit	NN	O	O
with	NN	O	O
simultaneous	NN	O	O
correction	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D	NN	O	O
deficiency	NN	O	O
were	NN	O	O
apparently	NN	O	O
responsible	NN	O	O
for	NN	O	O
high	NN	O	O
efficacy	NN	O	O
of	NN	O	O
the	NN	O	O
drug	NN	O	O
in	NN	O	O
treatment	NN	O	O
of	NN	O	O
calcium	NN	O	O
metabolism	NN	O	O
and	NN	O	O
immunity	NN	O	O
impairments	NN	O	O
in	NN	O	O
children	NN	O	O
with	NN	O	O
renal	NN	O	O
deteriorations	NN	O	O
at	NN	O	O
the	NN	O	O
step	NN	O	O
of	NN	O	O
chronic	NN	O	O
kidney	NN	O	O
insufficiency	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
The	NN	O	O
role	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
HLA	NN	O	B-protein
antigens	NN	O	I-protein
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
]	NN	O	O

Lymphocytic	NN	O	O
levels	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
were	NN	O	O
evaluated	NN	O	O
in	NN	O	O
114	NN	O	O
patients	NN	O	O
suffering	NN	O	O
from	NN	O	O
Icenko-Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Incidence	NN	O	O
of	NN	O	O
HLA	NN	O	B-protein
antigens	NN	O	I-protein
was	NN	O	O
determined	NN	O	O
in	NN	O	O
94	NN	O	O
of	NN	O	O
them	NN	O	O
.	NN	O	O

A	NN	O	O
significant	NN	O	O
rise	NN	O	O
of	NN	O	O
A10	NN	O	B-protein
and	NN	O	O
B27	NN	O	B-protein
antigen	NN	O	I-protein
incidence	NN	O	O
compared	NN	O	O
to	NN	O	O
that	NN	O	O
in	NN	O	O
normal	NN	O	O
subjects	NN	O	O
allows	NN	O	O
these	NN	O	O
antigens	NN	O	O
to	NN	O	O
be	NN	O	O
considered	NN	O	O
genetic	NN	O	O
markers	NN	O	O
of	NN	O	O
Icenko-Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
.	NN	O	O

The	NN	O	O
levels	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
patients	NN	O	O
are	NN	O	O
lower	NN	O	O
than	NN	O	O
in	NN	O	O
normal	NN	O	O
subjects	NN	O	O
both	NN	O	O
in	NN	O	O
the	NN	O	O
active	NN	O	O
stage	NN	O	O
of	NN	O	O
the	NN	O	O
disease	NN	O	O
and	NN	O	O
following	NN	O	O
bilateral	NN	O	O
total	NN	O	O
adrenalectomy	NN	O	O
.	NN	O	O

The	NN	O	O
patients	NN	O	O
carrying	NN	O	O
B27	NN	O	B-protein
antigen	NN	O	I-protein
had	NN	O	O
lymphocytic	NN	O	O
receptor	NN	O	O
concentrations	NN	O	O
under	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
such	NN	O	O
in	NN	O	O
patients	NN	O	O
free	NN	O	O
of	NN	O	O
the	NN	O	O
antigen	NN	O	O
carriage	NN	O	O
.	NN	O	O

Antigen	NN	O	B-protein
B27	NN	O	I-protein
seems	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
cause	NN	O	O
of	NN	O	O
lower	NN	O	O
levels	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
blood	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Effects	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
on	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
chronic	NN	O	I-cell_line
myelogenous	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
RWLeu-4	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
effects	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
on	NN	O	O
proliferation	NN	O	O
,	NN	O	O
differentiation	NN	O	O
,	NN	O	O
and	NN	O	O
macromolecular	NN	O	O
synthesis	NN	O	O
in	NN	O	O
the	NN	O	O
new	NN	O	O
Philadelphia	NN	O	B-cell_line
chromosome-positive	NN	O	I-cell_line
chronic	NN	O	I-cell_line
myelogenous	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
RWLeu-4	NN	O	B-cell_line
,	NN	O	O
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

Binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
VD3	NN	O	O
was	NN	O	O
saturable	NN	O	O
,	NN	O	O
with	NN	O	O
approximately	NN	O	O
2000-3000	NN	O	O
sites/cell	NN	O	O
,	NN	O	O
and	NN	O	O
half-maximal	NN	O	O
binding	NN	O	O
occurring	NN	O	O
at	NN	O	O
0.21-0.33	NN	O	O
nM	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
RWLeu-4	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
VD3	NN	O	O
induced	NN	O	O
24R-hydroxylase	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
a	NN	O	O
marker	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
responsiveness	NN	O	O
in	NN	O	O
many	NN	O	O
tissues	NN	O	O
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
RWLeu-4	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
VD3	NN	O	O
also	NN	O	O
inhibited	NN	O	O
proliferation	NN	O	O
and	NN	O	O
DNA	NN	O	O
synthesis	NN	O	O
with	NN	O	O
a	NN	O	O
50	NN	O	O
%	NN	O	O
effective	NN	O	O
dose	NN	O	O
of	NN	O	O
3.5-10	NN	O	O
nM	NN	O	O
within	NN	O	O
72	NN	O	O
h	NN	O	O
;	NN	O	O
in	NN	O	O
addition	NN	O	O
,	NN	O	O
protein	NN	O	O
and	NN	O	O
RNA	NN	O	O
synthesis	NN	O	O
were	NN	O	O
inhibited	NN	O	O
by	NN	O	O
VD3	NN	O	O
treatment	NN	O	O
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
RWLeu-4	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
5	NN	O	O
nM	NN	O	O
VD3	NN	O	O
for	NN	O	O
72	NN	O	O
h	NN	O	O
caused	NN	O	O
50	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
to	NN	O	O
differentiate	NN	O	O
into	NN	O	O
macrophage/monocyte	NN	O	B-cell_type
type	NN	O	I-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
judged	NN	O	O
by	NN	O	O
nitroblue	NN	O	O
tetrazolium	NN	O	O
staining	NN	O	O
and	NN	O	O
adherence	NN	O	O
to	NN	O	O
plastic	NN	O	O
.	NN	O	O

Progressive	NN	O	O
expression	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
maturation-specific	NN	O	I-protein
antigens	NN	O	I-protein
of	NN	O	O
the	NN	O	O
monocyte/macrophage	NN	O	B-cell_type
lineage	NN	O	I-cell_type
was	NN	O	O
induced	NN	O	O
by	NN	O	O
treatment	NN	O	O
of	NN	O	O
RWLeu-4	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
VD3	NN	O	O
for	NN	O	O
24	NN	O	O
to	NN	O	O
72	NN	O	O
h	NN	O	O
at	NN	O	O
doses	NN	O	O
that	NN	O	O
inhibited	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
.	NN	O	O

c-myc	NN	O	B-RNA
RNA	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
RWLeu-4	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
increased	NN	O	O
after	NN	O	O
0.5	NN	O	O
h	NN	O	O
of	NN	O	O
treatment	NN	O	O
with	NN	O	O
50	NN	O	O
nM	NN	O	O
VD3	NN	O	O
and	NN	O	O
then	NN	O	O
rapidly	NN	O	O
decreased	NN	O	O
to	NN	O	O
barely	NN	O	O
detectable	NN	O	O
levels	NN	O	O
after	NN	O	O
4	NN	O	O
h	NN	O	O
of	NN	O	O
treatment	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
the	NN	O	O
in	NN	O	O
vitro	NN	O	O
tyrosine	NN	O	O
kinase	NN	O	O
activity	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
p210bcr-abl	NN	O	B-protein
oncogene	NN	O	I-protein
product	NN	O	I-protein
was	NN	O	O
decreased	NN	O	O
approximately	NN	O	O
50	NN	O	O
%	NN	O	O
by	NN	O	O
VD3	NN	O	O
treatment	NN	O	O
.	NN	O	O

Because	NN	O	O
of	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
functional	NN	O	O
receptor-effector	NN	O	O
system	NN	O	O
for	NN	O	O
VD3	NN	O	O
and	NN	O	O
multiple	NN	O	O
biological	NN	O	O
responses	NN	O	O
to	NN	O	O
the	NN	O	O
hormone	NN	O	O
,	NN	O	O
these	NN	O	O
cells	NN	O	O
provide	NN	O	O
a	NN	O	O
unique	NN	O	O
model	NN	O	O
system	NN	O	O
with	NN	O	O
which	NN	O	O
to	NN	O	O
probe	NN	O	O
the	NN	O	O
specific	NN	O	O
effects	NN	O	O
of	NN	O	O
VD3	NN	O	O
on	NN	O	O
cell	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
in	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
new	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
leucine	NN	O	B-protein
zipper	NN	O	I-protein
class	NN	O	I-protein
of	NN	O	O
proteins	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
HLA	NN	O	B-DNA
DR	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Several	NN	O	O
mutants	NN	O	O
derived	NN	O	O
from	NN	O	O
transformed	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
are	NN	O	O
defective	NN	O	O
in	NN	O	O
expressing	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

The	NN	O	O
failure	NN	O	O
to	NN	O	O
express	NN	O	O
a	NN	O	O
class	NN	O	B-DNA
II	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
such	NN	O	O
mutant	NN	O	B-cell_line
line	NN	O	I-cell_line
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
MHC	NN	O	B-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
X	NN	O	I-DNA
box	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
conserved	NN	O	B-DNA
transcriptional	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
.	NN	O	O

A	NN	O	O
complementary	NN	O	B-DNA
DNA	NN	O	I-DNA
encoding	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
human	NN	O	B-protein
X	NN	O	I-protein
box	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
hXBP-1	NN	O	B-protein
)	NN	O	O
whose	NN	O	O
target	NN	O	O
is	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
DR	NN	O	I-DNA
alpha	NN	O	I-DNA
X	NN	O	I-DNA
box	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
region	NN	O	I-DNA
has	NN	O	O
now	NN	O	O
been	NN	O	O
cloned	NN	O	O
.	NN	O	O

This	NN	O	O
complementary	NN	O	B-DNA
DNA	NN	O	I-DNA
encoded	NN	O	O
a	NN	O	O
protein	NN	O	O
with	NN	O	O
structural	NN	O	O
similarities	NN	O	O
to	NN	O	O
the	NN	O	O
c-jun	NN	O	B-protein
proto-oncogene	NN	O	I-protein
product	NN	O	I-protein
,	NN	O	O
and	NN	O	O
its	NN	O	O
target	NN	O	O
sequence	NN	O	O
was	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
palindromic	NN	O	B-DNA
target	NN	O	I-DNA
sequence	NN	O	I-DNA
of	NN	O	O
c-jun	NN	O	B-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
hXBP-1	NN	O	B-DNA
DNA	NN	O	I-DNA
target	NN	O	I-DNA
sequence	NN	O	I-DNA
decreased	NN	O	O
DR	NN	O	B-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
vivo	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
hXBP-1	NN	O	B-protein
protein	NN	O	I-protein
acts	NN	O	O
as	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	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

Pseudohypoaldosteronism	NN	O	O
in	NN	O	O
eight	NN	O	O
families	NN	O	O
:	NN	O	O
different	NN	O	O
forms	NN	O	O
of	NN	O	O
inheritance	NN	O	O
are	NN	O	O
evidence	NN	O	O
for	NN	O	O
various	NN	O	O
genetic	NN	O	O
defects	NN	O	O
.	NN	O	O

Pseudohypoaldosteronism	NN	O	O
is	NN	O	O
a	NN	O	O
rare	NN	O	O
hereditary	NN	O	O
disorder	NN	O	O
presenting	NN	O	O
in	NN	O	O
early	NN	O	O
infancy	NN	O	O
with	NN	O	O
renal	NN	O	O
salt	NN	O	O
loss	NN	O	O
leading	NN	O	O
to	NN	O	O
hyponatremia	NN	O	O
and	NN	O	O
hyperkalemia	NN	O	O
despite	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
plasma	NN	O	O
aldosterone	NN	O	O
.	NN	O	O

The	NN	O	O
patients	NN	O	O
are	NN	O	O
insensitive	NN	O	O
to	NN	O	O
mineralocorticoids	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
sodium	NN	O	O
supplementation	NN	O	O
is	NN	O	O
able	NN	O	O
to	NN	O	O
correct	NN	O	O
electrolyte	NN	O	O
abnormalities	NN	O	O
.	NN	O	O

Absent	NN	O	O
or	NN	O	O
greatly	NN	O	O
diminished	NN	O	O
type	NN	O	O
I	NN	O	B-protein
aldosterone	NN	O	I-protein
receptors	NN	O	I-protein
in	NN	O	O
peripheral	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
have	NN	O	O
been	NN	O	O
recently	NN	O	O
demonstrated	NN	O	O
and	NN	O	O
explain	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
response	NN	O	O
to	NN	O	O
mineralocorticoids	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
studied	NN	O	O
the	NN	O	O
mode	NN	O	O
of	NN	O	O
inheritance	NN	O	O
in	NN	O	O
eight	NN	O	O
families	NN	O	O
with	NN	O	O
a	NN	O	O
total	NN	O	O
of	NN	O	O
nine	NN	O	O
patients	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
evidence	NN	O	O
for	NN	O	O
an	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
form	NN	O	O
of	NN	O	O
inheritance	NN	O	O
in	NN	O	O
four	NN	O	O
families	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
other	NN	O	O
four	NN	O	O
families	NN	O	O
appeared	NN	O	O
to	NN	O	O
have	NN	O	O
an	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
mode	NN	O	O
of	NN	O	O
transmission	NN	O	O
.	NN	O	O

In	NN	O	O
three	NN	O	O
families	NN	O	O
the	NN	O	O
autosomal	NN	O	O
recessive	NN	O	O
form	NN	O	O
was	NN	O	O
characterized	NN	O	O
by	NN	O	O
normal	NN	O	B-protein
receptor	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
hormone	NN	O	O
data	NN	O	O
in	NN	O	O
both	NN	O	O
parents	NN	O	O
,	NN	O	O
while	NN	O	O
in	NN	O	O
one	NN	O	O
family	NN	O	O
receptor	NN	O	O
levels	NN	O	O
in	NN	O	O
both	NN	O	O
parents	NN	O	O
were	NN	O	O
greatly	NN	O	O
reduced	NN	O	O
,	NN	O	O
but	NN	O	O
hormone	NN	O	O
levels	NN	O	O
were	NN	O	O
normal	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
four	NN	O	O
families	NN	O	O
with	NN	O	O
an	NN	O	O
autosomal	NN	O	O
dominant	NN	O	O
mode	NN	O	O
of	NN	O	O
transmission	NN	O	O
there	NN	O	O
was	NN	O	O
always	NN	O	O
one	NN	O	O
parent	NN	O	O
with	NN	O	O
reduced	NN	O	O
receptor	NN	O	O
binding	NN	O	O
in	NN	O	O
peripheral	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
and	NN	O	O
elevated	NN	O	O
serum	NN	O	O
hormone	NN	O	O
levels	NN	O	O
.	NN	O	O

These	NN	O	O
parents	NN	O	O
were	NN	O	O
entirely	NN	O	O
asymptomatic	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
extended	NN	O	O
family	NN	O	O
we	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
study	NN	O	O
an	NN	O	O
aunt	NN	O	O
and	NN	O	O
her	NN	O	O
newborn	NN	O	O
daughter	NN	O	O
,	NN	O	O
who	NN	O	O
were	NN	O	O
both	NN	O	O
also	NN	O	O
biochemically	NN	O	O
affected	NN	O	O
but	NN	O	O
clinically	NN	O	O
asymptomatic	NN	O	O
.	NN	O	O

It	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
appears	NN	O	O
that	NN	O	O
this	NN	O	O
dual	NN	O	O
pattern	NN	O	O
of	NN	O	O
genetic	NN	O	O
transmission	NN	O	O
may	NN	O	O
indicate	NN	O	O
differing	NN	O	O
genetic	NN	O	O
defects	NN	O	O
which	NN	O	O
cause	NN	O	O
the	NN	O	O
same	NN	O	O
clinical	NN	O	O
picture	NN	O	O
of	NN	O	O
pseudohypoaldosteronism	NN	O	O
.	NN	O	O

-DOCSTART-	O

Perceived	NN	O	O
social	NN	O	O
support	NN	O	O
and	NN	O	O
tumor	NN	O	O
estrogen/progesterone	NN	O	B-protein
receptor	NN	O	I-protein
status	NN	O	O
as	NN	O	O
predictors	NN	O	O
of	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cell	NN	O	I-cell_type
activity	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
.	NN	O	O

This	NN	O	O
report	NN	O	O
is	NN	O	O
concerned	NN	O	O
with	NN	O	O
the	NN	O	O
prediction	NN	O	O
of	NN	O	O
natural	NN	O	O
killer	NN	O	B-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
in	NN	O	O
61	NN	O	O
Stage	NN	O	O
I	NN	O	O
and	NN	O	O
II	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
,	NN	O	O
between	NN	O	O
the	NN	O	O
ages	NN	O	O
of	NN	O	O
25	NN	O	O
and	NN	O	O
70	NN	O	O
,	NN	O	O
who	NN	O	O
were	NN	O	O
accrued	NN	O	O
to	NN	O	O
this	NN	O	O
project	NN	O	O
.	NN	O	O

All	NN	O	O
baseline	NN	O	O
interview	NN	O	O
and	NN	O	O
testing	NN	O	O
data	NN	O	O
were	NN	O	O
obtained	NN	O	O
either	NN	O	O
just	NN	O	O
before	NN	O	O
patients	NN	O	O
were	NN	O	O
discharged	NN	O	O
from	NN	O	O
the	NN	O	O
hospital	NN	O	O
,	NN	O	O
or	NN	O	O
at	NN	O	O
their	NN	O	O
first	NN	O	O
outpatient	NN	O	O
visit	NN	O	O
,	NN	O	O
within	NN	O	O
two	NN	O	O
weeks	NN	O	O
of	NN	O	O
discharge	NN	O	O
.	NN	O	O

A	NN	O	O
major	NN	O	O
interest	NN	O	O
of	NN	O	O
this	NN	O	O
project	NN	O	O
is	NN	O	O
the	NN	O	O
predictive	NN	O	O
value	NN	O	O
of	NN	O	O
perceived	NN	O	O
social	NN	O	O
support	NN	O	O
,	NN	O	O
as	NN	O	O
a	NN	O	O
potential	NN	O	O
``	NN	O	O
stress	NN	O	O
''	NN	O	O
buffer	NN	O	O
,	NN	O	O
related	NN	O	O
to	NN	O	O
NK	NN	O	O
activity	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
main	NN	O	O
model	NN	O	O
reported	NN	O	O
here	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
a	NN	O	O
significant	NN	O	O
amount	NN	O	O
of	NN	O	O
NK	NN	O	O
activity	NN	O	O
variance	NN	O	O
could	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
five	NN	O	O
variables	NN	O	O
.	NN	O	O

Higher	NN	O	O
NK	NN	O	O
activity	NN	O	O
could	NN	O	O
be	NN	O	O
predicted	NN	O	O
by	NN	O	O
the	NN	O	O
perception	NN	O	O
of	NN	O	O
high	NN	O	O
quality	NN	O	O
emotional	NN	O	O
support	NN	O	O
from	NN	O	O
a	NN	O	O
spouse	NN	O	O
or	NN	O	O
intimate	NN	O	O
other	NN	O	O
,	NN	O	O
perceived	NN	O	O
social	NN	O	O
support	NN	O	O
from	NN	O	O
the	NN	O	O
patient	NN	O	O
's	NN	O	O
physician	NN	O	O
,	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
-negative	NN	O	O
tumor	NN	O	O
status	NN	O	O
,	NN	O	O
having	NN	O	O
an	NN	O	O
excisional	NN	O	O
biopsy	NN	O	O
as	NN	O	O
surgical	NN	O	O
treatment	NN	O	O
,	NN	O	O
and	NN	O	O
actively	NN	O	O
seeking	NN	O	O
social	NN	O	O
support	NN	O	O
as	NN	O	O
a	NN	O	O
major	NN	O	O
coping	NN	O	O
strategy	NN	O	O
(	NN	O	O
R2	NN	O	O
=	NN	O	O
0.33	NN	O	O
,	NN	O	O
F	NN	O	O
(	NN	O	O
5	NN	O	O
,	NN	O	O
55	NN	O	O
)	NN	O	O
=	NN	O	O
5.5	NN	O	O
,	NN	O	O
p	NN	O	O
less	NN	O	O
than	NN	O	O
0.0004	NN	O	O
)	NN	O	O
.	NN	O	O

Findings	NN	O	O
are	NN	O	O
discussed	NN	O	O
in	NN	O	O
terms	NN	O	O
of	NN	O	O
host	NN	O	O
interaction	NN	O	O
with	NN	O	O
tumor	NN	O	O
endocrine	NN	O	O
status	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
role	NN	O	O
that	NN	O	O
social	NN	O	O
support	NN	O	O
might	NN	O	O
play	NN	O	O
in	NN	O	O
modulating	NN	O	O
such	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Estrogen	NN	O	B-protein
receptor	NN	O	I-protein
content	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
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

ER	NN	O	O
content	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
from	NN	O	O
27	NN	O	O
SLE	NN	O	O
patients	NN	O	O
and	NN	O	O
20	NN	O	O
healthy	NN	O	O
controls	NN	O	O
were	NN	O	O
determined	NN	O	O
by	NN	O	O
dextran-coated	NN	O	O
charcoal	NN	O	O
assay	NN	O	O
.	NN	O	O

ER	NN	O	B-protein
content	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
each	NN	O	O
sample	NN	O	O
was	NN	O	O
expressed	NN	O	O
by	NN	O	O
both	NN	O	O
fmol/mg	NN	O	O
of	NN	O	O
lymphocyte	NN	O	B-protein
cytosolic	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
fmol/micrograms	NN	O	O
of	NN	O	O
lymphocyte	NN	O	B-DNA
DNA	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
significant	NN	O	O
difference	NN	O	O
between	NN	O	O
the	NN	O	O
ER	NN	O	B-protein
content	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
from	NN	O	O
the	NN	O	O
controls	NN	O	O
and	NN	O	O
that	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
SLE	NN	O	O
.	NN	O	O

But	NN	O	O
the	NN	O	O
logarithmic	NN	O	O
mean	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
content	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
expressed	NN	O	O
by	NN	O	O
fmol/mg	NN	O	O
of	NN	O	O
cytosolic	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
in	NN	O	O
14	NN	O	O
patients	NN	O	O
with	NN	O	O
active	NN	O	O
SLE	NN	O	O
(	NN	O	O
0.9356	NN	O	O
+/-	NN	O	O
0.31	NN	O	O
)	NN	O	O
was	NN	O	O
significantly	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
13	NN	O	O
patients	NN	O	O
with	NN	O	O
inactive	NN	O	O
SLE	NN	O	O
(	NN	O	O
0.2979	NN	O	O
+/-	NN	O	O
0.23	NN	O	O
,	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.001	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
controls	NN	O	O
(	NN	O	O
0.6204	NN	O	O
+/-	NN	O	O
0.52	NN	O	O
,	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

The	NN	O	O
normal	NN	O	O
upper	NN	O	O
limit	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
content	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
expressed	NN	O	O
by	NN	O	O
fmol/micrograms	NN	O	O
of	NN	O	O
DNA	NN	O	O
,	NN	O	O
was	NN	O	O
0.136	NN	O	O
.	NN	O	O

The	NN	O	O
elevated	NN	O	O
rate	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
content	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
in	NN	O	O
14	NN	O	O
active	NN	O	O
SLE	NN	O	O
(	NN	O	O
92.9	NN	O	O
%	NN	O	O
)	NN	O	O
was	NN	O	O
also	NN	O	O
higher	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
quieiescent	NN	O	O
patients	NN	O	O
(	NN	O	O
23.1	NN	O	O
%	NN	O	O
,	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.001	NN	O	O
)	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
controls	NN	O	O
(	NN	O	O
10	NN	O	O
%	NN	O	O
,	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

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
elevated	NN	O	O
level	NN	O	O
of	NN	O	O
ER	NN	O	B-protein
content	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
the	NN	O	O
positive	NN	O	O
antidsDNA	NN	O	B-protein
antibody	NN	O	I-protein
and	NN	O	O
hypocomplementemia	NN	O	O
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
in	NN	O	O
vitro	NN	O	O
globin	NN	O	B-DNA
gene	NN	O	I-DNA
switching	NN	O	O
model	NN	O	O
based	NN	O	O
on	NN	O	O
differentiated	NN	O	B-cell_type
embryonic	NN	O	I-cell_type
stem	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
used	NN	O	O
mouse	NN	O	B-cell_type
embryonic	NN	O	I-cell_type
stem	NN	O	I-cell_type
(	NN	O	I-cell_type
ES	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
study	NN	O	O
globin	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
and	NN	O	O
switching	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
ES-derived	NN	O	B-cell_type
embryoid	NN	O	I-cell_type
bodies	NN	O	I-cell_type
express	NN	O	O
the	NN	O	O
full	NN	O	O
complement	NN	O	O
of	NN	O	O
mouse	NN	O	B-DNA
embryonic	NN	O	I-DNA
globin	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
correct	NN	O	O
temporal	NN	O	O
order	NN	O	O
and	NN	O	O
that	NN	O	O
on	NN	O	O
further	NN	O	O
differentiation	NN	O	O
,	NN	O	O
a	NN	O	O
switch	NN	O	O
occurs	NN	O	O
to	NN	O	O
the	NN	O	O
fetal/adult	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
erythroid-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
NF-E1	NN	O	B-protein
was	NN	O	O
shown	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
coordinately	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
globin	NN	O	B-protein
in	NN	O	O
embryoid	NN	O	B-cell_type
bodies	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
from	NN	O	O
these	NN	O	O
experiments	NN	O	O
that	NN	O	O
the	NN	O	O
ES	NN	O	O
cell	NN	O	O
system	NN	O	O
provides	NN	O	O
a	NN	O	O
good	NN	O	O
model	NN	O	O
to	NN	O	O
study	NN	O	O
hematopoietic	NN	O	O
development	NN	O	O
.	NN	O	O

When	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
epsilon-	NN	O	I-DNA
or	NN	O	I-DNA
beta-globin	NN	O	I-DNA
genes	NN	O	I-DNA
driven	NN	O	O
by	NN	O	O
the	NN	O	O
dominant	NN	O	B-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
DCR	NN	O	B-DNA
)	NN	O	O
are	NN	O	O
introduced	NN	O	O
into	NN	O	O
this	NN	O	O
system	NN	O	O
,	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
epsilon-globin	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
beta-globin	NN	O	B-DNA
gene	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
not	NN	O	O
deregulated	NN	O	O
by	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
DCR	NN	O	B-DNA
and	NN	O	O
is	NN	O	O
expressed	NN	O	O
strictly	NN	O	O
as	NN	O	O
an	NN	O	O
embryonic	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
from	NN	O	O
this	NN	O	O
that	NN	O	O
the	NN	O	O
epsilon-globin	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
competition	NN	O	O
with	NN	O	O
other	NN	O	O
genes	NN	O	O
in	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

Cloning	NN	O	O
of	NN	O	O
a	NN	O	O
mitogen-inducible	NN	O	B-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
a	NN	O	O
kappa	NN	O	B-protein
B	NN	O	I-protein
DNA-binding	NN	O	I-protein
protein	NN	O	I-protein
with	NN	O	O
homology	NN	O	O
to	NN	O	O
the	NN	O	O
rel	NN	O	B-DNA
oncogene	NN	O	I-DNA
and	NN	O	O
to	NN	O	O
cell-cycle	NN	O	B-DNA
motifs	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
and	NN	O	O
characterized	NN	O	O
a	NN	O	O
mitogen-inducible	NN	O	B-DNA
gene	NN	O	I-DNA
isolated	NN	O	O
from	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
predicts	NN	O	O
a	NN	O	O
protein	NN	O	O
of	NN	O	O
968	NN	O	B-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
.	NN	O	O

The	NN	O	O
amino-terminal	NN	O	B-protein
domain	NN	O	I-protein
has	NN	O	O
regions	NN	O	O
homologous	NN	O	O
to	NN	O	O
the	NN	O	O
oncogene	NN	O	B-DNA
rel	NN	O	I-DNA
and	NN	O	O
to	NN	O	O
the	NN	O	O
developmentally	NN	O	O
important	NN	O	O
gene	NN	O	O
dorsal	NN	O	B-DNA
of	NN	O	O
Drosophila	NN	O	O
.	NN	O	O

The	NN	O	O
carboxy-terminal	NN	O	B-protein
domain	NN	O	I-protein
contains	NN	O	O
repeat	NN	O	O
structures	NN	O	O
found	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
proteins	NN	O	O
that	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
cell-cycle	NN	O	O
control	NN	O	O
of	NN	O	O
yeast	NN	O	O
and	NN	O	O
in	NN	O	O
tissue	NN	O	O
differentiation	NN	O	O
in	NN	O	O
Drosophila	NN	O	O
and	NN	O	O
Ceanorhabditis	NN	O	O
elegans	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
putative	NN	O	B-DNA
human	NN	O	I-DNA
oncogene	NN	O	I-DNA
bcl-3	NN	O	I-DNA
and	NN	O	O
in	NN	O	O
the	NN	O	O
ankyrin	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

A	NN	O	O
truncated	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
product	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
translated	NN	O	O
in	NN	O	O
vitro	NN	O	O
is	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
protein	NN	O	I-protein
which	NN	O	O
interacts	NN	O	O
specifically	NN	O	O
with	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
found	NN	O	O
in	NN	O	O
many	NN	O	O
inducible	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
including	NN	O	O
the	NN	O	O
enhancer	NN	O	O
in	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
.	NN	O	O

This	NN	O	O
gene	NN	O	O
is	NN	O	O
yet	NN	O	O
another	NN	O	O
in	NN	O	O
a	NN	O	O
growing	NN	O	O
list	NN	O	O
of	NN	O	O
important	NN	O	O
regulatory	NN	O	O
molecules	NN	O	O
whose	NN	O	O
expression	NN	O	O
is	NN	O	O
transcriptionally	NN	O	O
induced	NN	O	O
upon	NN	O	O
cellular	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Extrarenal	NN	O	O
receptor-effector-mechanisms	NN	O	O
for	NN	O	O
aldosterone	NN	O	O
:	NN	O	O
the	NN	O	O
sequence	NN	O	O
of	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
cellular	NN	O	O
electrolyte	NN	O	O
transport	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
their	NN	O	O
implications	NN	O	O
for	NN	O	O
disorders	NN	O	O
of	NN	O	O
the	NN	O	O
water	NN	O	O
and	NN	O	O
electrolyte	NN	O	O
balances	NN	O	O
.	NN	O	O

High	NN	O	O
affinity	NN	O	O
aldosterone	NN	O	O
binding	NN	O	O
sites	NN	O	O
have	NN	O	O
not	NN	O	O
only	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
the	NN	O	O
classic	NN	O	O
target	NN	O	O
tissues	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
renal	NN	O	O
tubules	NN	O	O
,	NN	O	O
but	NN	O	O
also	NN	O	O
in	NN	O	O
non-classic	NN	O	O
target	NN	O	O
tissues	NN	O	O
such	NN	O	O
as	NN	O	O
the	NN	O	O
hippocampus	NN	O	O
,	NN	O	O
mammary	NN	O	O
gland	NN	O	O
,	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
,	NN	O	O
recently	NN	O	O
,	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
.	NN	O	O

An	NN	O	O
in	NN	O	O
vitro	NN	O	O
effect	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
on	NN	O	O
intracellular	NN	O	O
sodium	NN	O	O
,	NN	O	O
potassium	NN	O	O
and	NN	O	O
calcium	NN	O	O
concentrations	NN	O	O
and	NN	O	O
cell	NN	O	O
volume	NN	O	O
was	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
.	NN	O	O

In	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
,	NN	O	O
the	NN	O	O
intracellular	NN	O	O
Na+	NN	O	O
,	NN	O	O
K+	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
concentrations	NN	O	O
and	NN	O	O
the	NN	O	O
cell	NN	O	O
volume	NN	O	O
decreased	NN	O	O
significantly	NN	O	O
,	NN	O	O
but	NN	O	O
remained	NN	O	O
constant	NN	O	O
when	NN	O	O
aldosterone	NN	O	O
(	NN	O	O
1.4	NN	O	O
nmol/l	NN	O	O
)	NN	O	O
was	NN	O	O
added	NN	O	O
to	NN	O	O
the	NN	O	O
incubation	NN	O	O
medium	NN	O	O
.	NN	O	O

These	NN	O	O
effects	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
were	NN	O	O
blocked	NN	O	O
by	NN	O	O
the	NN	O	O
aldosterone	NN	O	O
antagonist	NN	O	O
canrenone	NN	O	O
(	NN	O	O
140	NN	O	O
nmol/l	NN	O	O
)	NN	O	O
.	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
could	NN	O	O
be	NN	O	O
identified	NN	O	O
as	NN	O	O
the	NN	O	O
primary	NN	O	O
target	NN	O	O
of	NN	O	O
the	NN	O	O
aldosterone	NN	O	O
action	NN	O	O
,	NN	O	O
possibly	NN	O	O
non-genomically	NN	O	O
mediated	NN	O	O
through	NN	O	O
membrane	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
significance	NN	O	O
of	NN	O	O
this	NN	O	O
model	NN	O	O
was	NN	O	O
underlined	NN	O	O
by	NN	O	O
the	NN	O	O
demonstration	NN	O	O
of	NN	O	O
absent	NN	O	O
or	NN	O	O
a	NN	O	O
decreased	NN	O	O
number	NN	O	O
of	NN	O	O
mineralocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
electrolyte	NN	O	O
response	NN	O	O
to	NN	O	O
aldosterone	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
of	NN	O	O
patients	NN	O	O
with	NN	O	O
pseudohypoaldosteronism	NN	O	O
and	NN	O	O
aldosteronism	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
an	NN	O	O
abnormal	NN	O	O
effector	NN	O	O
mechanism	NN	O	O
could	NN	O	O
be	NN	O	O
demonstrated	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
from	NN	O	O
essential	NN	O	O
hypertensives	NN	O	O
.	NN	O	O

These	NN	O	O
studies	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
significance	NN	O	O
of	NN	O	O
extrarenal	NN	O	B-protein
,	NN	O	I-protein
nonepithelial	NN	O	I-protein
mineralocorticoid	NN	O	I-protein
receptors	NN	O	I-protein
and	NN	O	O
the	NN	O	O
related	NN	O	O
effector	NN	O	O
mechanism	NN	O	O
in	NN	O	O
different	NN	O	O
disorders	NN	O	O
of	NN	O	O
the	NN	O	O
water	NN	O	O
and	NN	O	O
electrolyte	NN	O	O
balance	NN	O	O
in	NN	O	O
man	NN	O	O
.	NN	O	O

-DOCSTART-	O

Immunohistochemical	NN	O	O
study	NN	O	O
of	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
and	NN	O	O
an	NN	O	O
estrogen	NN	O	O
binding	NN	O	O
assay	NN	O	O
in	NN	O	O
malignant	NN	O	B-cell_type
soft	NN	O	I-cell_type
tissue	NN	O	I-cell_type
tumors	NN	O	I-cell_type
.	NN	O	O

Immunohistochemically	NN	O	O
,	NN	O	O
the	NN	O	O
immunoreaction	NN	O	O
against	NN	O	O
5	NN	O	O
steroid	NN	O	O
hormone	NN	O	O
anti-sera	NN	O	O
(	NN	O	O
estradiol	NN	O	O
,	NN	O	O
estriol	NN	O	O
,	NN	O	O
cortisol	NN	O	O
,	NN	O	O
progesterone	NN	O	O
and	NN	O	O
testosterone	NN	O	O
)	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
39	NN	O	O
cases	NN	O	O
with	NN	O	O
the	NN	O	O
malignant	NN	O	B-cell_type
soft	NN	O	I-cell_type
tissue	NN	O	I-cell_type
tumors	NN	O	I-cell_type
(	NN	O	O
fibrosarcoma	NN	O	O
:	NN	O	O
8	NN	O	O
,	NN	O	O
malignant	NN	O	O
fibrous	NN	O	O
histiocytoma	NN	O	O
:	NN	O	O
6	NN	O	O
,	NN	O	O
rhabdomyosarcoma	NN	O	O
:	NN	O	O
10	NN	O	O
,	NN	O	O
leiomyosarcoma	NN	O	O
:	NN	O	O
10	NN	O	O
,	NN	O	O
liposarcoma	NN	O	O
:	NN	O	O
5	NN	O	O
)	NN	O	O
.	NN	O	O

Seventeen	NN	O	O
cases	NN	O	O
revealed	NN	O	O
distinct	NN	O	O
immunostaining	NN	O	O
against	NN	O	O
at	NN	O	O
least	NN	O	O
1	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
.	NN	O	O

Immunostained	NN	O	B-cell_type
tumor	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
more	NN	O	O
frequently	NN	O	O
distributed	NN	O	O
in	NN	O	O
the	NN	O	O
area	NN	O	O
where	NN	O	O
tumor	NN	O	O
cell	NN	O	O
infiltration	NN	O	O
was	NN	O	O
more	NN	O	O
invasive	NN	O	O
.	NN	O	O

The	NN	O	O
majority	NN	O	O
of	NN	O	O
the	NN	O	O
positive	NN	O	O
cases	NN	O	O
occurred	NN	O	O
in	NN	O	O
female	NN	O	O
cases	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
estrogen	NN	O	O
binding	NN	O	O
activity	NN	O	O
)	NN	O	O
was	NN	O	O
examined	NN	O	O
histochemically	NN	O	O
in	NN	O	O
39	NN	O	O
cases	NN	O	O
and	NN	O	O
it	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
8	NN	O	O
.	NN	O	O

We	NN	O	O
concluded	NN	O	O
that	NN	O	O
steroid	NN	O	O
hormones	NN	O	O
might	NN	O	O
be	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
tumor	NN	O	O
cell	NN	O	O
infiltration	NN	O	O
of	NN	O	O
some	NN	O	O
malignant	NN	O	B-cell_type
soft	NN	O	I-cell_type
tissue	NN	O	I-cell_type
tumors	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
cyclic	NN	O	B-protein
AMP-dependent	NN	O	I-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
in	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathway	NN	O	O
for	NN	O	O
interleukin-1	NN	O	B-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
a	NN	O	O
highly	NN	O	O
specific	NN	O	O
protein	NN	O	O
inhibitor	NN	O	O
for	NN	O	O
cyclic	NN	O	B-protein
AMP-dependent	NN	O	I-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
in	NN	O	O
interleukin-1	NN	O	B-cell_line
(	NN	O	I-cell_line
IL-1	NN	O	I-cell_line
)	NN	O	I-cell_line
-responsive	NN	O	I-cell_line
cells	NN	O	I-cell_line
blocked	NN	O	O
IL-1-induced	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
that	NN	O	O
was	NN	O	O
driven	NN	O	O
by	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
immunoglobulin	NN	O	I-DNA
enhancer	NN	O	I-DNA
or	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

This	NN	O	O
inhibitor	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
-mediated	NN	O	O
gene	NN	O	O
transcription	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
cyclic	NN	O	B-protein
AMP-dependent	NN	O	I-protein
protein	NN	O	I-protein
kinases	NN	O	I-protein
are	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
for	NN	O	O
IL-1	NN	O	B-protein
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
responsive	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	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
ORI1yt	NN	O	I-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
not	NN	O	O
B-cell	NN	O	O
specific	NN	O	O
and	NN	O	O
does	NN	O	O
not	NN	O	O
respond	NN	O	O
synergistically	NN	O	O
to	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
R	NN	O	I-protein
and	NN	O	I-protein
Z	NN	O	I-protein
.	NN	O	O

The	NN	O	O
Epstein-Barr	NN	O	B-DNA
virus	NN	O	I-DNA
DR	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
PstI	NN	O	B-DNA
repeats	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
it	NN	O	O
contains	NN	O	O
an	NN	O	O
upstream	NN	O	B-DNA
region	NN	O	I-DNA
(	NN	O	O
positions	NN	O	O
-69	NN	O	O
to	NN	O	O
-220	NN	O	O
)	NN	O	O
responsive	NN	O	O
to	NN	O	O
EB1	NN	O	B-protein
(	NN	O	I-protein
Z	NN	O	I-protein
)	NN	O	I-protein
(	NN	O	O
the	NN	O	O
BZLF1-encoded	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
)	NN	O	O
and	NN	O	O
an	NN	O	O
enhancer	NN	O	B-DNA
with	NN	O	O
two	NN	O	O
functionally	NN	O	O
distinct	NN	O	O
domains	NN	O	O
,	NN	O	O
A	NN	O	O
and	NN	O	O
B	NN	O	B-DNA
.	NN	O	O

Domain	NN	O	B-DNA
B	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
described	NN	O	O
as	NN	O	O
a	NN	O	O
B-cell-specific	NN	O	B-DNA
EB1-responsive	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
P.M.Lieberman	NN	O	O
,	NN	O	O
J.M.Hardwick	NN	O	O
,	NN	O	O
and	NN	O	O
S.D.Hayward	NN	O	O
,	NN	O	O
J.Virol.63	NN	O	O
:	NN	O	O
3040-3050	NN	O	O
,	NN	O	O
1989	NN	O	O
)	NN	O	O
activated	NN	O	O
synergistically	NN	O	O
by	NN	O	O
EB1	NN	O	B-protein
and	NN	O	O
R	NN	O	B-protein
,	NN	O	O
an	NN	O	O
EBV	NN	O	B-protein
early	NN	O	I-protein
product	NN	O	I-protein
encoded	NN	O	O
by	NN	O	O
the	NN	O	O
open	NN	O	B-DNA
reading	NN	O	I-DNA
frame	NN	O	I-DNA
BRLF1	NN	O	I-DNA
(	NN	O	O
M.A.	NN	O	O
Cox	NN	O	O
,	NN	O	O
J.Leahy	NN	O	O
,	NN	O	O
and	NN	O	O
J.M.Hardwick	NN	O	O
,	NN	O	O
J.Virol.64	NN	O	O
:	NN	O	O
313-321	NN	O	O
,	NN	O	O
1990	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
domain	NN	O	B-DNA
B	NN	O	I-DNA
is	NN	O	O
an	NN	O	O
R-responsive	NN	O	B-DNA
element	NN	O	I-DNA
in	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
is	NN	O	O
therefore	NN	O	O
not	NN	O	O
an	NN	O	O
EB1-responsive	NN	O	B-DNA
B-cell-specific	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
an	NN	O	O
EB1-binding	NN	O	B-DNA
site	NN	O	I-DNA
(	NN	O	O
ZRE-B	NN	O	B-DNA
)	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
R-responsive	NN	O	B-DNA
enhancer	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

ZRE-B	NN	O	B-DNA
can	NN	O	O
be	NN	O	O
deleted	NN	O	O
without	NN	O	O
affecting	NN	O	O
the	NN	O	O
R-dependent	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
no	NN	O	O
cooperation	NN	O	O
or	NN	O	O
synergy	NN	O	O
between	NN	O	O
R	NN	O	B-protein
and	NN	O	O
EB1	NN	O	B-protein
when	NN	O	O
activating	NN	O	O
the	NN	O	O
B	NN	O	B-DNA
domain	NN	O	I-DNA
(	NN	O	O
ZRE-B	NN	O	B-DNA
plus	NN	O	O
the	NN	O	O
R-responsive	NN	O	B-DNA
element	NN	O	I-DNA
)	NN	O	O
positioned	NN	O	O
as	NN	O	O
an	NN	O	O
enhancer	NN	O	B-DNA
.	NN	O	O

ZRE-B	NN	O	B-DNA
is	NN	O	O
therefore	NN	O	O
not	NN	O	O
part	NN	O	O
of	NN	O	O
the	NN	O	O
R-	NN	O	B-DNA
inducible	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
have	NN	O	O
tested	NN	O	O
several	NN	O	O
subregions	NN	O	O
of	NN	O	O
the	NN	O	O
DR	NN	O	B-DNA
enhancer	NN	O	I-DNA
B	NN	O	B-DNA
domain	NN	O	I-DNA
,	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
for	NN	O	O
their	NN	O	O
capacity	NN	O	O
to	NN	O	O
transmit	NN	O	O
the	NN	O	O
R-activating	NN	O	O
signal	NN	O	O
to	NN	O	O
the	NN	O	O
rabbit	NN	O	B-DNA
beta-globin	NN	O	I-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
R-responsive	NN	O	B-DNA
element	NN	O	I-DNA
is	NN	O	O
composed	NN	O	O
of	NN	O	O
four	NN	O	O
protoenhancers	NN	O	B-DNA
that	NN	O	O
span	NN	O	O
the	NN	O	O
whole	NN	O	O
B	NN	O	B-DNA
domain	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
protoenhancers	NN	O	B-DNA
alone	NN	O	O
are	NN	O	O
weakly	NN	O	O
or	NN	O	O
not	NN	O	O
responsive	NN	O	O
to	NN	O	O
R	NN	O	B-protein
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
protoenhancers	NN	O	B-DNA
contains	NN	O	O
the	NN	O	O
overlapping	NN	O	B-DNA
palindromes	NN	O	I-DNA
5'-TTGTCCcgtGGACAAaTGTCC-3'	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
one	NN	O	O
palindrome	NN	O	B-DNA
,	NN	O	O
either	NN	O	O
alone	NN	O	O
or	NN	O	O
duplicated	NN	O	O
,	NN	O	O
or	NN	O	O
the	NN	O	O
overlapping	NN	O	B-DNA
palindromes	NN	O	I-DNA
did	NN	O	O
not	NN	O	O
respond	NN	O	O
to	NN	O	O
R	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	O
3	NN	O	B-protein
,	NN	O	I-protein
5	NN	O	I-protein
,	NN	O	I-protein
3'-triiodothyronine	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
T3R	NN	O	B-protein
)	NN	O	O
of	NN	O	O
circulating	NN	O	B-cell_type
human	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
hyper-	NN	O	O
and	NN	O	O
hypothyroidism	NN	O	O
and	NN	O	O
nonthyroidal	NN	O	O
diseases	NN	O	O
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
implications	NN	O	O
of	NN	O	O
nuclear	NN	O	O
T3R	NN	O	B-protein
alterations	NN	O	O
of	NN	O	O
circulating	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
hyperthyroidism	NN	O	O
,	NN	O	O
hypothyroidism	NN	O	O
and	NN	O	O
nonthyroidal	NN	O	O
diseases	NN	O	O
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

Nuclear	NN	O	O
T3R	NN	O	B-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
was	NN	O	O
determined	NN	O	O
by	NN	O	O
radio-ligand	NN	O	O
binding	NN	O	O
analysis	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
in	NN	O	O
hyper-	NN	O	O
and	NN	O	O
hypothyroid	NN	O	O
patients	NN	O	O
the	NN	O	O
nuclear	NN	O	O
affinity	NN	O	O
(	NN	O	O
Ka	NN	O	O
)	NN	O	O
for	NN	O	O
T3	NN	O	O
was	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
normal	NN	O	O
subjects	NN	O	O
.	NN	O	O

In	NN	O	O
hyperthyroidism	NN	O	O
nuclear	NN	O	O
T3	NN	O	O
maximal	NN	O	O
binding	NN	O	O
capacity	NN	O	O
(	NN	O	O
MBC	NN	O	O
)	NN	O	O
was	NN	O	O
unaltered	NN	O	O
,	NN	O	O
whereas	NN	O	O
in	NN	O	O
hypothyroidism	NN	O	O
the	NN	O	O
MBC	NN	O	O
was	NN	O	O
significantly	NN	O	O
increased	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
patients	NN	O	O
with	NN	O	O
diabetes	NN	O	O
mellitus	NN	O	O
,	NN	O	O
chronic	NN	O	O
renal	NN	O	O
failure	NN	O	O
and	NN	O	O
hepatic	NN	O	O
cirrhosis	NN	O	O
,	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
T3R	NN	O	I-protein
MBC	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
was	NN	O	O
about	NN	O	O
1.5-1.6	NN	O	O
times	NN	O	O
of	NN	O	O
the	NN	O	O
normal	NN	O	O
controls	NN	O	O
.	NN	O	O

It	NN	O	O
was	NN	O	O
concluded	NN	O	O
that	NN	O	O
there	NN	O	O
existed	NN	O	O
hormonal	NN	O	O
regulation	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
T3R	NN	O	I-protein
,	NN	O	O
and	NN	O	O
up-regulation	NN	O	O
was	NN	O	O
seen	NN	O	O
in	NN	O	O
hypothyroidism	NN	O	O
and	NN	O	O
low	NN	O	O
T3	NN	O	O
syndrome	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lymphocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
from	NN	O	O
vitamin	NN	O	O
D-dependent	NN	O	O
rickets	NN	O	O
type	NN	O	O
II	NN	O	O
show	NN	O	O
functional	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	I-protein
25-dihydroxyvitamin	NN	O	I-protein
D3	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Lymphocyte	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
from	NN	O	O
five	NN	O	O
patients	NN	O	O
with	NN	O	O
vitamin	NN	O	O
D-dependent	NN	O	O
rickets	NN	O	O
,	NN	O	O
type	NN	O	O
II	NN	O	O
(	NN	O	O
VDDR-II	NN	O	O
)	NN	O	O
.	NN	O	O

These	NN	O	O
lines	NN	O	O
were	NN	O	O
established	NN	O	O
by	NN	O	O
infection	NN	O	O
with	NN	O	O
human	NN	O	O
T-lymphotrophic	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

Binding	NN	O	O
of	NN	O	O
[	NN	O	O
3H	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
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
to	NN	O	O
its	NN	O	O
receptor	NN	O	O
in	NN	O	O
these	NN	O	O
cell	NN	O	O
lines	NN	O	O
was	NN	O	O
compared	NN	O	O
to	NN	O	O
binding	NN	O	O
studies	NN	O	O
using	NN	O	O
a	NN	O	O
T-lymphocyte	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
S-LB1	NN	O	B-cell_line
)	NN	O	O
from	NN	O	O
a	NN	O	O
normal	NN	O	O
individual	NN	O	O
.	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
receptor	NN	O	O
of	NN	O	O
S-LB1	NN	O	B-cell_line
was	NN	O	O
comparable	NN	O	O
to	NN	O	O
the	NN	O	O
well-characterized	NN	O	O
chick	NN	O	B-protein
intestinal	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
receptor	NN	O	I-protein
in	NN	O	O
terms	NN	O	O
of	NN	O	O
its	NN	O	O
ligand	NN	O	O
binding	NN	O	O
affinity	NN	O	O
and	NN	O	O
capacity	NN	O	O
,	NN	O	O
its	NN	O	O
mobility	NN	O	O
on	NN	O	O
5-20	NN	O	O
%	NN	O	O
sucrose	NN	O	O
gradients	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
adsorption	NN	O	O
to	NN	O	O
and	NN	O	O
elution	NN	O	O
properties	NN	O	O
from	NN	O	O
DNA-cellulose	NN	O	O
.	NN	O	O

Three	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
VDDR-II	NN	O	O
(	NN	O	O
Rh-	NN	O	O
VDR	NN	O	B-protein
,	NN	O	O
Sh-	NN	O	O
VDR	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Ab-	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
showed	NN	O	O
no	NN	O	O
specific	NN	O	O
binding	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
a	NN	O	O
receptor	NN	O	B-protein
and	NN	O	O
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
cultured	NN	O	B-cell_line
cells	NN	O	I-cell_line
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
did	NN	O	O
not	NN	O	O
stimulate	NN	O	O
production	NN	O	O
of	NN	O	O
24	NN	O	O
,	NN	O	O
25-dihydroxy-vitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
24	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
a	NN	O	O
response	NN	O	O
which	NN	O	O
is	NN	O	O
diagnostic	NN	O	O
of	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
functional	NN	O	B-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
receptor	NN	O	I-protein
.	NN	O	O

In	NN	O	O
a	NN	O	O
fourth	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
A1-VDR	NN	O	B-cell_line
,	NN	O	O
the	NN	O	O
receptor	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
had	NN	O	O
a	NN	O	O
low	NN	O	O
binding	NN	O	O
capacity	NN	O	O
and	NN	O	O
25	NN	O	B-protein
(	NN	O	I-protein
OH	NN	O	I-protein
)	NN	O	I-protein
D3-24-hydroxylase	NN	O	I-protein
activity	NN	O	O
was	NN	O	O
not	NN	O	O
detectable	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
24	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
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
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
fifth	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
designated	NN	O	O
Ro-VDR	NN	O	B-cell_line
,	NN	O	O
although	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
hormone	NN	O	O
treatment	NN	O	O
was	NN	O	O
lower	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
from	NN	O	O
a	NN	O	O
normal	NN	O	O
donor	NN	O	O
.	NN	O	O

The	NN	O	O
capacity	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
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
was	NN	O	O
low	NN	O	O
in	NN	O	O
Ro-VDR	NN	O	B-cell_line
.	NN	O	O

In	NN	O	O
all	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
where	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
binding	NN	O	O
to	NN	O	O
a	NN	O	O
receptor	NN	O	B-protein
was	NN	O	O
detectable	NN	O	O
,	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
had	NN	O	O
the	NN	O	O
typical	NN	O	O
sedimentation	NN	O	O
coefficient	NN	O	O
of	NN	O	O
3.7	NN	O	O
S	NN	O	O
on	NN	O	O
sucrose	NN	O	O
density	NN	O	O
gradient	NN	O	O
analysis	NN	O	O
.	NN	O	O

Binding	NN	O	O
and	NN	O	O
elution	NN	O	O
properties	NN	O	O
to	NN	O	O
DNA-cellulose	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
differed	NN	O	O
from	NN	O	O
normal	NN	O	O
in	NN	O	O
both	NN	O	O
Ro-VDR	NN	O	B-cell_line
and	NN	O	O
A1-VDR	NN	O	B-cell_line
cells	NN	O	I-cell_line
where	NN	O	O
elution	NN	O	O
from	NN	O	O
DNA-cellulose	NN	O	O
occurred	NN	O	O
at	NN	O	O
a	NN	O	O
lower	NN	O	O
salt	NN	O	O
concentration	NN	O	O
than	NN	O	O
is	NN	O	O
typical	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
.	NN	O	O

While	NN	O	O
Ro-VDR	NN	O	B-cell_line
cells	NN	O	I-cell_line
showed	NN	O	O
typical	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
of	NN	O	O
the	NN	O	O
unoccupied	NN	O	B-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
receptor	NN	O	I-protein
,	NN	O	O
neither	NN	O	O
the	NN	O	O
unoccupied	NN	O	O
nor	NN	O	O
the	NN	O	O
occupied	NN	O	O
receptor	NN	O	B-protein
from	NN	O	O
A1-VDR	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
completely	NN	O	O
localized	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
series	NN	O	O
of	NN	O	O
functional	NN	O	O
studies	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
modulation	NN	O	O
of	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
mRNAs	NN	O	B-RNA
coding	NN	O	O
for	NN	O	O
both	NN	O	O
the	NN	O	O
c-myc	NN	O	B-DNA
oncogene	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
growth	NN	O	O
factor	NN	O	O
known	NN	O	O
as	NN	O	O
granulocyte-monocyte	NN	O	O
colony	NN	O	O
stimulating	NN	O	O
activity	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
correlated	NN	O	O
with	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
status	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
.	NN	O	O

Use	NN	O	O
of	NN	O	O
these	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
will	NN	O	O
facilitate	NN	O	O
further	NN	O	O
study	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
defect	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
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
in	NN	O	O
vitamin	NN	O	O
D-dependent	NN	O	O
rickets	NN	O	O
type	NN	O	O
II	NN	O	O
and	NN	O	O
will	NN	O	O
allow	NN	O	O
a	NN	O	O
correlation	NN	O	O
with	NN	O	O
impairment	NN	O	O
of	NN	O	O
cellular	NN	O	O
functions	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
and	NN	O	O
post-transcriptional	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
human	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
leukemic	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

AP-1	NN	O	B-protein
,	NN	O	O
the	NN	O	O
polypeptide	NN	O	O
product	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	O
recognizes	NN	O	O
and	NN	O	O
binds	NN	O	O
to	NN	O	O
specific	NN	O	O
DNA	NN	O	O
sequences	NN	O	O
and	NN	O	O
stimulates	NN	O	O
transcription	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
responsive	NN	O	O
to	NN	O	O
certain	NN	O	O
growth	NN	O	O
factors	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-tetradecanoylphorbol-13-acetate	NN	O	O
(	NN	O	O
TPA	NN	O	O
)	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
TPA	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
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
during	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Low	NN	O	O
levels	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
transcripts	NN	O	I-RNA
were	NN	O	O
detectable	NN	O	O
in	NN	O	O
untreated	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
leukemic	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
increased	NN	O	O
significantly	NN	O	O
by	NN	O	O
6	NN	O	O
h	NN	O	O
,	NN	O	O
and	NN	O	O
reached	NN	O	O
near	NN	O	O
maximal	NN	O	O
levels	NN	O	O
by	NN	O	O
24	NN	O	O
h	NN	O	O
of	NN	O	O
exposure	NN	O	O
to	NN	O	O
32	NN	O	O
nM	NN	O	O
TPA	NN	O	O
.	NN	O	O

Similar	NN	O	O
kinetics	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
induction	NN	O	O
by	NN	O	O
TPA	NN	O	O
were	NN	O	O
observed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
U-937	NN	O	I-cell_line
and	NN	O	O
THP-1	NN	O	B-cell_line
monocytic	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Similar	NN	O	O
findings	NN	O	O
were	NN	O	O
obtained	NN	O	O
with	NN	O	O
bryostatin	NN	O	O
1	NN	O	O
(	NN	O	O
10	NN	O	O
nM	NN	O	O
)	NN	O	O
,	NN	O	O
another	NN	O	O
activator	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
inducer	NN	O	O
of	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Furthermore	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
0.5	NN	O	O
microM	NN	O	O
)	NN	O	O
,	NN	O	O
a	NN	O	O
structurally	NN	O	O
distinct	NN	O	O
agent	NN	O	O
which	NN	O	O
also	NN	O	O
induces	NN	O	O
HL-60	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
,	NN	O	O
increased	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
.	NN	O	O

TPA	NN	O	O
treatment	NN	O	O
of	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
cycloheximide	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
superinduction	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
transcripts	NN	O	O
.	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
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
untreated	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
that	NN	O	O
exposure	NN	O	O
to	NN	O	O
TPA	NN	O	O
increases	NN	O	O
this	NN	O	O
rate	NN	O	O
3.3-fold	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
both	NN	O	O
TPA	NN	O	O
and	NN	O	O
cycloheximide	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
the	NN	O	O
rates	NN	O	O
of	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
RNA	NN	O	I-RNA
as	NN	O	O
determined	NN	O	O
by	NN	O	O
treating	NN	O	O
HL-60	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
TPA	NN	O	O
and	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
was	NN	O	O
30	NN	O	O
min	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	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
RNA	NN	O	I-RNA
in	NN	O	O
TPA-treated	NN	O	B-cell_line
HL-60	NN	O	I-cell_line
cells	NN	O	I-cell_line
exposed	NN	O	O
to	NN	O	O
cycloheximide	NN	O	O
and	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
was	NN	O	O
greater	NN	O	O
than	NN	O	O
2	NN	O	O
h	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
c-jun	NN	O	B-RNA
RNA	NN	O	I-RNA
observed	NN	O	O
during	NN	O	O
TPA-induced	NN	O	O
monocytic	NN	O	O
differentiation	NN	O	O
is	NN	O	O
mediated	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

-DOCSTART-	O

[	NN	O	O
Hormonal	NN	O	O
interactions	NN	O	O
and	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
the	NN	O	O
nephrotic	NN	O	O
syndrome	NN	O	O
]	NN	O	O

As	NN	O	O
many	NN	O	O
as	NN	O	O
27	NN	O	O
children	NN	O	O
aged	NN	O	O
6	NN	O	O
to	NN	O	O
15	NN	O	O
years	NN	O	O
with	NN	O	O
morphologically	NN	O	O
verified	NN	O	O
nephropathies	NN	O	O
were	NN	O	O
examined	NN	O	O
.	NN	O	O

Four	NN	O	O
variants	NN	O	O
of	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
thyroid	NN	O	O
status	NN	O	O
,	NN	O	O
characteristic	NN	O	O
of	NN	O	O
children	NN	O	O
with	NN	O	O
different	NN	O	O
variants	NN	O	O
of	NN	O	O
nephrotic	NN	O	O
syndrome	NN	O	O
were	NN	O	O
distinguished	NN	O	O
:	NN	O	O
1	NN	O	O
)	NN	O	O
biochemical	NN	O	O
signs	NN	O	O
of	NN	O	O
primary	NN	O	O
hypothyroidism	NN	O	O
,	NN	O	O
2	NN	O	O
)	NN	O	O
biochemical	NN	O	O
signs	NN	O	O
of	NN	O	O
secondary	NN	O	O
hypothyroidism	NN	O	O
,	NN	O	O
3	NN	O	O
)	NN	O	O
low	NN	O	O
content	NN	O	O
of	NN	O	O
T3	NN	O	O
,	NN	O	O
4	NN	O	O
)	NN	O	O
dysfunction	NN	O	O
of	NN	O	O
the	NN	O	O
hypophyseal	NN	O	O
and	NN	O	O
thyroid	NN	O	O
system	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
steroid	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
thyroid	NN	O	O
hormones	NN	O	O
that	NN	O	O
the	NN	O	O
low	NN	O	O
level	NN	O	O
of	NN	O	O
steroid	NN	O	B-protein
receptors	NN	O	I-protein
,	NN	O	O
thyroid	NN	O	O
hormones	NN	O	O
(	NN	O	O
T3	NN	O	O
and	NN	O	O
T4	NN	O	O
)	NN	O	O
and	NN	O	O
cortisol	NN	O	O
is	NN	O	O
typical	NN	O	O
of	NN	O	O
children	NN	O	O
with	NN	O	O
the	NN	O	O
signs	NN	O	O
of	NN	O	O
renal	NN	O	O
dysplasia	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
assumed	NN	O	O
that	NN	O	O
superaddition	NN	O	O
under	NN	O	O
such	NN	O	O
conditions	NN	O	O
of	NN	O	O
immune	NN	O	O
glomerulopathy	NN	O	O
(	NN	O	O
glomerulonephritis	NN	O	O
and	NN	O	O
nephrotic	NN	O	O
syndrome	NN	O	O
)	NN	O	O
gives	NN	O	O
rise	NN	O	O
to	NN	O	O
the	NN	O	O
resistance	NN	O	O
to	NN	O	O
the	NN	O	O
treatment	NN	O	O
with	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

-DOCSTART-	O

Lymphoid	NN	O	O
specific	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
adenovirus	NN	O	B-DNA
early	NN	O	I-DNA
region	NN	O	I-DNA
3	NN	O	I-DNA
promoter	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
primary	NN	O	O
site	NN	O	O
of	NN	O	O
infection	NN	O	O
by	NN	O	O
human	NN	O	O
adenoviruses	NN	O	O
is	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

However	NN	O	O
,	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
control	NN	O	I-DNA
elements	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
adenoviral	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
has	NN	O	O
not	NN	O	O
been	NN	O	O
reported	NN	O	O
.	NN	O	O

The	NN	O	O
adenovirus	NN	O	B-protein
early	NN	O	I-protein
region	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	I-protein
ES	NN	O	I-protein
)	NN	O	I-protein
gene	NN	O	I-protein
products	NN	O	I-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
maintenance	NN	O	O
of	NN	O	O
viral	NN	O	O
persistence	NN	O	O
by	NN	O	O
complexing	NN	O	O
with	NN	O	O
the	NN	O	O
class	NN	O	B-protein
I	NN	O	I-protein
MHC	NN	O	I-protein
antigens	NN	O	I-protein
,	NN	O	O
thus	NN	O	O
preventing	NN	O	O
their	NN	O	O
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
with	NN	O	O
a	NN	O	O
resultant	NN	O	O
decrease	NN	O	O
in	NN	O	O
host	NN	O	O
immunologic	NN	O	O
destruction	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
different	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
were	NN	O	O
involved	NN	O	O
in	NN	O	O
E3	NN	O	O
regulation	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
as	NN	O	O
compared	NN	O	O
with	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
both	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
transfection	NN	O	O
analysis	NN	O	O
with	NN	O	O
the	NN	O	O
E3	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
both	NN	O	O
cell	NN	O	O
types	NN	O	O
were	NN	O	O
performed	NN	O	O
.	NN	O	O

These	NN	O	O
studies	NN	O	O
detected	NN	O	O
two	NN	O	O
novel	NN	O	B-DNA
domains	NN	O	I-DNA
referred	NN	O	O
to	NN	O	O
as	NN	O	O
L1	NN	O	B-DNA
and	NN	O	O
L2	NN	O	B-DNA
with	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
lymphoid	NN	O	O
but	NN	O	O
not	NN	O	O
HeLa	NN	O	O
extracts	NN	O	O
.	NN	O	O

Each	NN	O	O
of	NN	O	O
these	NN	O	O
domains	NN	O	O
possessed	NN	O	O
strong	NN	O	O
homology	NN	O	O
to	NN	O	O
motifs	NN	O	O
previously	NN	O	O
found	NN	O	O
to	NN	O	O
bind	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Transfections	NN	O	O
of	NN	O	O
E3	NN	O	B-DNA
constructs	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
gene	NN	O	I-DNA
revealed	NN	O	O
that	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
motif	NN	O	I-DNA
(	NN	O	O
L2	NN	O	B-DNA
)	NN	O	O
had	NN	O	O
minimal	NN	O	O
effects	NN	O	O
on	NN	O	O
promoter	NN	O	O
expression	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
resulted	NN	O	O
in	NN	O	O
dramatic	NN	O	O
decreases	NN	O	O
in	NN	O	O
expression	NN	O	O
by	NN	O	O
lymphoid	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
mutagenesis	NN	O	O
of	NN	O	O
proximal	NN	O	B-DNA
NF-kappa	NN	O	I-DNA
B	NN	O	I-DNA
motif	NN	O	I-DNA
(	NN	O	O
L1	NN	O	B-DNA
)	NN	O	O
had	NN	O	O
minimal	NN	O	O
effects	NN	O	O
on	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
both	NN	O	O
HeLa	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
but	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
small	NN	O	O
,	NN	O	O
but	NN	O	O
reproducible	NN	O	O
,	NN	O	O
increase	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
when	NN	O	O
coupled	NN	O	O
to	NN	O	O
the	NN	O	O
L2	NN	O	B-DNA
mutation	NN	O	O
.	NN	O	O

Reversing	NN	O	O
the	NN	O	O
position	NN	O	O
and	NN	O	O
subsequent	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
L1	NN	O	B-DNA
and	NN	O	O
L2	NN	O	B-DNA
domains	NN	O	I-DNA
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
primary	NN	O	B-DNA
sequence	NN	O	I-DNA
of	NN	O	O
these	NN	O	O
motifs	NN	O	O
rather	NN	O	O
than	NN	O	O
their	NN	O	O
position	NN	O	O
in	NN	O	O
the	NN	O	O
E3	NN	O	B-DNA
promoter	NN	O	I-DNA
was	NN	O	O
critical	NN	O	O
for	NN	O	O
regulating	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

Characterization	NN	O	O
of	NN	O	O
defensin	NN	O	B-protein
resistance	NN	O	O
phenotypes	NN	O	O
associated	NN	O	O
with	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
phoP	NN	O	B-DNA
virulence	NN	O	I-DNA
regulon	NN	O	I-DNA
of	NN	O	O
Salmonella	NN	O	O
typhimurium	NN	O	O
.	NN	O	O

The	NN	O	O
defensin	NN	O	B-protein
sensitivities	NN	O	O
of	NN	O	O
Salmonella	NN	O	O
typhimurium	NN	O	O
strains	NN	O	O
with	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
phoP/phoQ	NN	O	B-DNA
two-component	NN	O	I-DNA
virulence	NN	O	I-DNA
regulon	NN	O	I-DNA
were	NN	O	O
tested	NN	O	O
by	NN	O	O
using	NN	O	O
purified	NN	O	B-protein
defensins	NN	O	I-protein
NP-1	NN	O	B-protein
and	NN	O	O
NP-2	NN	O	B-protein
.	NN	O	O

Strains	NN	O	O
with	NN	O	O
mutations	NN	O	O
in	NN	O	O
either	NN	O	O
gene	NN	O	O
of	NN	O	O
the	NN	O	O
regulatory	NN	O	B-DNA
pair	NN	O	I-DNA
(	NN	O	O
phoP	NN	O	B-DNA
[	NN	O	O
transcriptional	NN	O	B-DNA
activator	NN	O	I-DNA
]	NN	O	O
or	NN	O	O
phoQ	NN	O	B-DNA
[	NN	O	O
membrane	NN	O	B-protein
sensor	NN	O	I-protein
kinase	NN	O	I-protein
]	NN	O	O
)	NN	O	O
had	NN	O	O
increased	NN	O	O
sensitivities	NN	O	O
to	NN	O	O
defensin	NN	O	B-protein
.	NN	O	O

The	NN	O	O
predicted	NN	O	O
periplasmic	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
PhoQ	NN	O	B-protein
protein	NN	O	I-protein
contained	NN	O	O
a	NN	O	O
markedly	NN	O	O
anionic	NN	O	O
domain	NN	O	O
that	NN	O	O
could	NN	O	O
interact	NN	O	O
with	NN	O	O
cationic	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
that	NN	O	O
could	NN	O	O
be	NN	O	O
responsible	NN	O	O
for	NN	O	O
resistance	NN	O	O
to	NN	O	O
defensin	NN	O	B-protein
.	NN	O	O

Because	NN	O	O
insertion	NN	O	O
mutations	NN	O	O
in	NN	O	O
phoP	NN	O	B-DNA
are	NN	O	O
polar	NN	O	O
on	NN	O	O
phoQ	NN	O	B-DNA
,	NN	O	O
we	NN	O	O
constructed	NN	O	O
strains	NN	O	O
that	NN	O	O
expressed	NN	O	O
the	NN	O	O
PhoQ	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
PhoP	NN	O	B-protein
to	NN	O	O
test	NN	O	O
whether	NN	O	O
resistance	NN	O	O
to	NN	O	O
defensin	NN	O	O
requires	NN	O	O
only	NN	O	O
the	NN	O	O
phoQ	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
resistance	NN	O	O
to	NN	O	O
defensin	NN	O	O
requires	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
both	NN	O	O
components	NN	O	O
of	NN	O	O
this	NN	O	O
regulatory	NN	O	O
system	NN	O	O
,	NN	O	O
because	NN	O	O
strains	NN	O	O
expressing	NN	O	O
PhoQ	NN	O	B-protein
without	NN	O	O
PhoP	NN	O	B-protein
were	NN	O	O
still	NN	O	O
markedly	NN	O	O
sensitive	NN	O	O
to	NN	O	O
defensins	NN	O	B-protein
.	NN	O	O

This	NN	O	O
implied	NN	O	O
that	NN	O	O
a	NN	O	O
pag	NN	O	B-protein
(	NN	O	I-protein
phoP-activated	NN	O	I-protein
gene	NN	O	I-protein
)	NN	O	I-protein
product	NN	O	I-protein
is	NN	O	O
responsible	NN	O	O
for	NN	O	O
defensin	NN	O	B-protein
resistance	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
tested	NN	O	O
for	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
defensins	NN	O	B-protein
NP-1	NN	O	B-protein
,	NN	O	O
NP-5	NN	O	B-protein
,	NN	O	O
and	NN	O	O
HNP-1	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
pag	NN	O	B-DNA
expression	NN	O	O
and	NN	O	O
found	NN	O	O
that	NN	O	O
these	NN	O	O
peptides	NN	O	O
have	NN	O	O
no	NN	O	O
effect	NN	O	O
.	NN	O	O

Defensin	NN	O	O
resistance	NN	O	O
is	NN	O	O
not	NN	O	O
the	NN	O	O
only	NN	O	O
virulence	NN	O	O
characteristic	NN	O	O
controlled	NN	O	O
by	NN	O	O
the	NN	O	O
PhoP-PhoQ	NN	O	B-protein
regulon	NN	O	I-protein
because	NN	O	O
mutations	NN	O	O
in	NN	O	O
pagC	NN	O	B-DNA
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
ones	NN	O	O
in	NN	O	O
the	NN	O	O
phoP	NN	O	B-DNA
locus	NN	O	I-DNA
that	NN	O	O
resulted	NN	O	O
in	NN	O	O
constitutive	NN	O	O
pag	NN	O	B-DNA
activation	NN	O	O
(	NN	O	O
phenotype	NN	O	O
PhoPc	NN	O	B-protein
)	NN	O	O
,	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
defensin	NN	O	B-protein
resistance	NN	O	O
,	NN	O	O
even	NN	O	O
though	NN	O	O
they	NN	O	O
rendered	NN	O	O
the	NN	O	O
organism	NN	O	O
avirulent	NN	O	O
and	NN	O	O
deficient	NN	O	O
in	NN	O	O
survival	NN	O	O
within	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

The	NN	O	O
virulence	NN	O	O
defect	NN	O	O
conferred	NN	O	O
by	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
phoP	NN	O	B-DNA
-phoQ	NN	O	B-DNA
two-component	NN	O	O
regulatory	NN	O	O
system	NN	O	O
is	NN	O	O
not	NN	O	O
completely	NN	O	O
explained	NN	O	O
by	NN	O	O
alterations	NN	O	O
in	NN	O	O
resistance	NN	O	O
to	NN	O	O
cationic	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
involves	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
other	NN	O	O
proteins	NN	O	O
necessary	NN	O	O
for	NN	O	O
S.	NN	O	O
typhimurium	NN	O	O
survival	NN	O	O
within	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Stimulation	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
clone	NN	O	I-cell_line
with	NN	O	O
anti-CD3	NN	O	B-protein
or	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
induces	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
but	NN	O	O
not	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
1	NN	O	O
enhancer-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
transiently	NN	O	O
transfected	NN	O	O
expression	NN	O	O
vectors	NN	O	O
under	NN	O	O
the	NN	O	O
control	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
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
or	NN	O	O
its	NN	O	O
enhancer	NN	O	B-DNA
sequence	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV	NN	O	B-protein
enhancer-binding	NN	O	I-protein
protein	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
were	NN	O	O
analyzed	NN	O	O
in	NN	O	O
two	NN	O	O
human	NN	O	O
T-cell	NN	O	B-cell_line
clones	NN	O	I-cell_line
stimulated	NN	O	O
through	NN	O	O
their	NN	O	O
T-cell	NN	O	B-protein
receptor	NN	O	I-protein
complex	NN	O	I-protein
or	NN	O	O
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
or	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
a	NN	O	O
dissociation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
from	NN	O	O
transactivation	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
or	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Interleukin	NN	O	B-protein
2	NN	O	I-protein
induced	NN	O	O
proliferation	NN	O	O
but	NN	O	O
not	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
or	NN	O	O
LTR	NN	O	B-DNA
transactivation	NN	O	O
.	NN	O	O

Phorbol	NN	O	O
ester	NN	O	O
or	NN	O	O
specific	NN	O	O
antigen	NN	O	O
recognition	NN	O	O
induced	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
transactivation	NN	O	O
,	NN	O	O
whereas	NN	O	O
stimulation	NN	O	O
with	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
or	NN	O	O
antibody	NN	O	O
to	NN	O	O
CD3	NN	O	B-protein
did	NN	O	O
not	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
latter	NN	O	O
signals	NN	O	O
were	NN	O	O
nevertheless	NN	O	O
able	NN	O	O
to	NN	O	O
induce	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
translocation	NN	O	O
with	NN	O	O
a	NN	O	O
pattern	NN	O	O
in	NN	O	O
the	NN	O	O
band-shift	NN	O	O
assay	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
that	NN	O	O
observed	NN	O	O
using	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
.	NN	O	O

Our	NN	O	O
finding	NN	O	O
that	NN	O	O
induction	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
by	NN	O	O
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
or	NN	O	O
antibody	NN	O	B-protein
to	NN	O	I-protein
CD3	NN	O	I-protein
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
induce	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
-dependent	NN	O	O
transcription	NN	O	O
in	NN	O	O
cloned	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
contrasts	NN	O	O
with	NN	O	O
results	NN	O	O
obtained	NN	O	O
in	NN	O	O
most	NN	O	O
lymphoblastoid	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
indicates	NN	O	O
that	NN	O	O
normal	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
differ	NN	O	O
from	NN	O	O
tumoral	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
terms	NN	O	O
of	NN	O	O
requirements	NN	O	O
for	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
activation	NN	O	O
.	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
events	NN	O	O
linked	NN	O	O
to	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
,	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
translocation	NN	O	O
per	NN	O	O
se	NN	O	O
,	NN	O	O
induce	NN	O	O
functional	NN	O	O
interactions	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
with	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Decreased	NN	O	O
concentration	NN	O	O
of	NN	O	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
mononuclear	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
X-linked	NN	O	O
hypophosphatemic	NN	O	O
rickets	NN	O	O
:	NN	O	O
effect	NN	O	O
of	NN	O	O
phosphate	NN	O	O
supplementation	NN	O	O
.	NN	O	O

Abnormal	NN	O	O
renal	NN	O	O
tubular	NN	O	O
phosphate	NN	O	O
transport	NN	O	O
is	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
primary	NN	O	O
defect	NN	O	O
in	NN	O	O
X-linked	NN	O	O
hypophosphatemic	NN	O	O
rickets	NN	O	O
(	NN	O	O
XLH	NN	O	O
)	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
resistance	NN	O	O
to	NN	O	O
vitamin	NN	O	O
D	NN	O	O
treatment	NN	O	O
in	NN	O	O
XLH	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
hypophosphatemia	NN	O	O
alone	NN	O	O
.	NN	O	O

Since	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
its	NN	O	O
receptors	NN	O	O
(	NN	O	O
VDR	NN	O	B-protein
)	NN	O	O
,	NN	O	O
abnormalities	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
have	NN	O	O
been	NN	O	O
postulated	NN	O	O
in	NN	O	O
XLH	NN	O	O
.	NN	O	O

In	NN	O	O
order	NN	O	O
to	NN	O	O
investigate	NN	O	O
this	NN	O	O
possibility	NN	O	O
,	NN	O	O
we	NN	O	O
measured	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
in	NN	O	O
PHA-activated	NN	O	B-cell_line
peripheral	NN	O	I-cell_line
mononuclear	NN	O	I-cell_line
cells	NN	O	I-cell_line
from	NN	O	O
10	NN	O	O
XLH	NN	O	O
patients	NN	O	O
.	NN	O	O

Patients	NN	O	O
without	NN	O	O
phosphate	NN	O	O
supplementation	NN	O	O
showed	NN	O	O
significantly	NN	O	O
lower	NN	O	O
concentration	NN	O	O
(	NN	O	O
21.7	NN	O	O
+/-	NN	O	O
5.1	NN	O	O
fmol/mg	NN	O	O
protein	NN	O	O
,	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SEM	NN	O	O
)	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
normal	NN	O	O
controls	NN	O	O
(	NN	O	O
60.7	NN	O	O
+/-	NN	O	O
4.0	NN	O	O
)	NN	O	O
.	NN	O	O

On	NN	O	O
the	NN	O	O
contrary	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
no	NN	O	O
significant	NN	O	O
difference	NN	O	O
between	NN	O	O
the	NN	O	O
phosphate-supplemented	NN	O	O
patients	NN	O	O
(	NN	O	O
58.3	NN	O	O
+/-	NN	O	O
2.7	NN	O	O
)	NN	O	O
and	NN	O	O
controls	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
significant	NN	O	O
positive	NN	O	O
correlation	NN	O	O
between	NN	O	O
VDR	NN	O	B-protein
concentration	NN	O	O
and	NN	O	O
serum	NN	O	O
phosphate	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

In	NN	O	O
two	NN	O	O
patients	NN	O	O
,	NN	O	O
VDR	NN	O	B-protein
was	NN	O	O
increased	NN	O	O
after	NN	O	O
daily	NN	O	O
phosphate	NN	O	O
supplementation	NN	O	O
was	NN	O	O
started	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
decreased	NN	O	O
concentration	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
secondary	NN	O	O
to	NN	O	O
persistent	NN	O	O
hypophosphatemia	NN	O	O
is	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
causes	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D	NN	O	O
resistance	NN	O	O
in	NN	O	O
XLH	NN	O	O
.	NN	O	O

-DOCSTART-	O

Two	NN	O	O
distinct	NN	O	O
forms	NN	O	O
of	NN	O	O
active	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
CREB	NN	O	I-protein
(	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
.	NN	O	O

Mammalian	NN	O	B-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
two	NN	O	O
distinct	NN	O	O
forms	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
CREB	NN	O	I-protein
(	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
that	NN	O	O
are	NN	O	O
apparently	NN	O	O
the	NN	O	O
products	NN	O	O
of	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
of	NN	O	O
the	NN	O	O
CREB	NN	O	B-RNA
gene	NN	O	I-RNA
transcript	NN	O	I-RNA
.	NN	O	O

The	NN	O	O
two	NN	O	O
proteins	NN	O	O
differ	NN	O	O
by	NN	O	O
a	NN	O	O
14-amino	NN	O	B-protein
acid	NN	O	I-protein
serine-rich	NN	O	I-protein
insertion	NN	O	I-protein
present	NN	O	O
in	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
CREB	NN	O	B-protein
isoforms	NN	O	I-protein
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
both	NN	O	O
CREB	NN	O	B-protein
isoforms	NN	O	I-protein
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
many	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
and	NN	O	O
mammalian	NN	O	O
species	NN	O	O
.	NN	O	O

Both	NN	O	O
encode	NN	O	O
proteins	NN	O	O
that	NN	O	O
bind	NN	O	O
specifically	NN	O	O
to	NN	O	O
a	NN	O	O
cAMP	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	I-DNA
vitro	NN	O	I-DNA
.	NN	O	O

As	NN	O	O
expected	NN	O	O
for	NN	O	O
proteins	NN	O	O
of	NN	O	O
this	NN	O	O
class	NN	O	O
,	NN	O	O
the	NN	O	O
CREB	NN	O	B-protein
proteins	NN	O	I-protein
bind	NN	O	O
DNA	NN	O	O
as	NN	O	O
dimers	NN	O	O
.	NN	O	O

Both	NN	O	O
proteins	NN	O	O
impart	NN	O	O
cAMP-regulated	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
to	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
DNA-binding	NN	O	I-DNA
domain	NN	O	I-DNA
,	NN	O	O
showing	NN	O	O
that	NN	O	O
cAMP	NN	O	O
directly	NN	O	O
modulates	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
stimulatory	NN	O	O
activity	NN	O	O
of	NN	O	O
CREB	NN	O	B-protein
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
multiple	NN	O	O
CREB	NN	O	B-protein
isoforms	NN	O	I-protein
with	NN	O	O
identical	NN	O	O
DNA-binding	NN	O	O
specificities	NN	O	O
but	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
presumed	NN	O	O
regulatory	NN	O	B-protein
domain	NN	O	I-protein
raises	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
CREB	NN	O	B-protein
proteins	NN	O	I-protein
may	NN	O	O
be	NN	O	O
able	NN	O	O
to	NN	O	O
integrate	NN	O	O
distinct	NN	O	O
regulatory	NN	O	O
signals	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

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
vpr	NN	O	O
product	NN	O	O
is	NN	O	O
a	NN	O	O
virion-associated	NN	O	B-protein
regulatory	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
vpr	NN	O	B-protein
product	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	NN	O	O
1	NN	O	O
(	NN	O	O
HIV-1	NN	O	O
)	NN	O	O
acts	NN	O	O
in	NN	O	O
trans	NN	O	O
to	NN	O	O
accelerate	NN	O	O
virus	NN	O	O
replication	NN	O	O
and	NN	O	O
cytopathic	NN	O	O
effect	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
it	NN	O	O
is	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
HIV-1	NN	O	O
viral	NN	O	O
particle	NN	O	O
contains	NN	O	O
multiple	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
vpr	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
vpr	NN	O	B-protein
product	NN	O	I-protein
is	NN	O	O
the	NN	O	O
first	NN	O	O
regulatory	NN	O	O
protein	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
to	NN	O	O
be	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
virus	NN	O	O
particle	NN	O	O
.	NN	O	O

This	NN	O	O
observation	NN	O	O
raises	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
vpr	NN	O	B-DNA
acts	NN	O	O
to	NN	O	O
facilitate	NN	O	O
the	NN	O	O
early	NN	O	O
steps	NN	O	O
of	NN	O	O
infection	NN	O	O
before	NN	O	O
de	NN	O	O
novo	NN	O	O
viral	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
occurs	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
B-cell	NN	O	B-protein
lineage-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
present	NN	O	O
at	NN	O	O
early	NN	O	O
but	NN	O	O
not	NN	O	O
late	NN	O	O
stages	NN	O	O
of	NN	O	O
differentiation	NN	O	O
.	NN	O	O

A	NN	O	O
novel	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
was	NN	O	O
identified	NN	O	O
as	NN	O	O
a	NN	O	O
mammalian	NN	O	B-protein
homolog	NN	O	I-protein
of	NN	O	O
the	NN	O	O
sea	NN	O	B-protein
urchin	NN	O	I-protein
protein	NN	O	I-protein
TSAP	NN	O	B-protein
,	NN	O	O
which	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
four	NN	O	O
tissue-specific	NN	O	B-DNA
late	NN	O	I-DNA
histone	NN	O	I-DNA
H2A-2	NN	O	I-DNA
and	NN	O	I-DNA
H2B-2	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

As	NN	O	O
shown	NN	O	O
by	NN	O	O
mobility-shift	NN	O	O
,	NN	O	O
methylation	NN	O	O
interference	NN	O	O
,	NN	O	O
and	NN	O	O
mutational	NN	O	O
analyses	NN	O	O
,	NN	O	O
the	NN	O	O
mammalian	NN	O	B-protein
protein	NN	O	I-protein
BSAP	NN	O	B-protein
recognizes	NN	O	O
all	NN	O	O
four	NN	O	O
sea	NN	O	B-DNA
urchin	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
manner	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
TSAP	NN	O	B-protein
;	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
two	NN	O	O
proteins	NN	O	O
differ	NN	O	O
in	NN	O	O
molecular	NN	O	O
weight	NN	O	O
.	NN	O	O

BSAP	NN	O	B-protein
is	NN	O	O
exclusively	NN	O	O
restricted	NN	O	O
to	NN	O	O
the	NN	O	O
B-cell	NN	O	O
lineage	NN	O	O
of	NN	O	O
lymphoid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Its	NN	O	O
expression	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
activated	NN	O	O
during	NN	O	O
pro-B-cell	NN	O	O
development	NN	O	O
,	NN	O	O
is	NN	O	O
abundant	NN	O	O
at	NN	O	O
the	NN	O	O
pre-B-	NN	O	O
and	NN	O	O
mature	NN	O	O
B-cell	NN	O	O
stages	NN	O	O
,	NN	O	O
but	NN	O	O
is	NN	O	O
absent	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

Moreover	NN	O	O
,	NN	O	O
BSAP	NN	O	B-protein
is	NN	O	O
clearly	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
as	NN	O	O
a	NN	O	O
wild-type	NN	O	B-DNA
but	NN	O	O
not	NN	O	O
a	NN	O	O
mutant	NN	O	B-DNA
TSAP-binding	NN	O	I-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
sea	NN	O	O
urchin	NN	O	O
functions	NN	O	O
only	NN	O	O
in	NN	O	O
transfected	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
an	NN	O	O
upstream	NN	O	B-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

Competition	NN	O	O
experiments	NN	O	O
did	NN	O	O
not	NN	O	O
reveal	NN	O	O
any	NN	O	O
high-affinity	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
for	NN	O	O
BSAP	NN	O	B-protein
in	NN	O	O
known	NN	O	O
regulatory	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
immunoglobulin	NN	O	B-DNA
and	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
major	NN	O	I-DNA
histocompatibility	NN	O	I-DNA
(	NN	O	I-DNA
MHC	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
BSAP	NN	O	B-protein
is	NN	O	O
a	NN	O	O
regulator	NN	O	O
of	NN	O	O
a	NN	O	O
different	NN	O	O
set	NN	O	O
of	NN	O	O
B-lymphoid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Octamer	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
and	NN	O	O
the	NN	O	O
cell	NN	O	B-DNA
type-specificity	NN	O	I-DNA
of	NN	O	I-DNA
immunoglobulin	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

Antibodies	NN	O	B-protein
are	NN	O	O
produced	NN	O	O
exclusively	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
expression	NN	O	O
of	NN	O	O
the	NN	O	O
antibody-encoding	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
the	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
,	NN	O	O
is	NN	O	O
also	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

The	NN	O	O
octamer	NN	O	B-DNA
sequence	NN	O	I-DNA
ATGCAAAT	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
of	NN	O	O
Ig	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
its	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
.	NN	O	O

This	NN	O	O
sequence	NN	O	B-DNA
motif	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
the	NN	O	O
so-called	NN	O	O
octamer	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
(	NN	O	O
Oct	NN	O	B-protein
or	NN	O	O
OTF	NN	O	B-protein
factors	NN	O	I-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
Oct-1	NN	O	B-protein
protein	NN	O	I-protein
is	NN	O	O
present	NN	O	O
in	NN	O	O
all	NN	O	O
cell	NN	O	O
types	NN	O	O
analyzed	NN	O	O
so	NN	O	O
far	NN	O	O
,	NN	O	O
whereas	NN	O	O
Oct-2A	NN	O	B-protein
and	NN	O	O
Oct-2B	NN	O	B-protein
are	NN	O	O
found	NN	O	O
mainly	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

All	NN	O	O
three	NN	O	O
proteins	NN	O	O
show	NN	O	O
the	NN	O	O
same	NN	O	O
sequence	NN	O	O
specificity	NN	O	O
and	NN	O	O
binding	NN	O	O
affinity	NN	O	O
.	NN	O	O

It	NN	O	O
appears	NN	O	O
that	NN	O	O
the	NN	O	O
B	NN	O	O
cell-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
Ig	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
mediated	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
cell	NN	O	O
type-specific	NN	O	B-protein
Oct	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
both	NN	O	O
quantitative	NN	O	O
and	NN	O	O
qualitative	NN	O	O
differences	NN	O	O
between	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
other	NN	O	O
octamer	NN	O	B-protein
factor	NN	O	I-protein
variants	NN	O	I-protein
were	NN	O	O
identified	NN	O	O
.	NN	O	O

Many	NN	O	O
of	NN	O	O
these	NN	O	O
may	NN	O	O
be	NN	O	O
created	NN	O	O
by	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
of	NN	O	O
a	NN	O	O
primary	NN	O	B-RNA
transcript	NN	O	I-RNA
of	NN	O	O
one	NN	O	O
Oct	NN	O	O
factor	NN	O	O
gene	NN	O	O
and	NN	O	O
may	NN	O	O
serve	NN	O	O
a	NN	O	O
specific	NN	O	O
function	NN	O	O
in	NN	O	O
the	NN	O	O
fine	NN	O	O
tuning	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
factor	NN	O	O
known	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
endogenous	NN	O	B-DNA
Ig	NN	O	I-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
only	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
is	NN	O	O
a	NN	O	O
ubiquitously	NN	O	B-protein
active	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
transcriptional	NN	O	B-DNA
enhancer	NN	O	I-DNA
located	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	O
intron	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
constant	NN	O	I-DNA
region	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
major	NN	O	O
determinant	NN	O	O
of	NN	O	O
B-cell-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

Like	NN	O	O
other	NN	O	O
enhancers	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
contains	NN	O	O
several	NN	O	O
short	NN	O	O
sequence	NN	O	B-DNA
motifs	NN	O	I-DNA
that	NN	O	O
bind	NN	O	O
specific	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

Each	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
overall	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
,	NN	O	O
however	NN	O	O
no	NN	O	O
single	NN	O	O
element	NN	O	O
seems	NN	O	O
absolutely	NN	O	O
required	NN	O	O
for	NN	O	O
activity	NN	O	O
.	NN	O	O

For	NN	O	O
a	NN	O	O
better	NN	O	O
understanding	NN	O	O
of	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
components	NN	O	I-DNA
,	NN	O	O
we	NN	O	O
have	NN	O	O
cloned	NN	O	O
and	NN	O	O
analyzed	NN	O	O
individual	NN	O	O
sequence	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
the	NN	O	O
factor	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
E3	NN	O	B-DNA
enhancer	NN	O	I-DNA
motif	NN	O	I-DNA
,	NN	O	O
CATGTGGC	NN	O	O
,	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
.	NN	O	O

It	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
an	NN	O	O
active	NN	O	O
form	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
non-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
where	NN	O	O
it	NN	O	O
can	NN	O	O
mediate	NN	O	O
transcriptional	NN	O	O
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

However	NN	O	O
,	NN	O	O
despite	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
a	NN	O	O
transfected	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
factor	NN	O	O
is	NN	O	O
apparently	NN	O	O
unable	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
Ig	NN	O	I-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
non-lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
In	NN	O	O
previous	NN	O	O
experiments	NN	O	O
by	NN	O	O
others	NN	O	O
,	NN	O	O
the	NN	O	O
characteristic	NN	O	O
in	NN	O	O
vivo	NN	O	O
footprint	NN	O	O
of	NN	O	O
this	NN	O	O
factor	NN	O	O
,	NN	O	O
designated	NN	O	O
NF-muE3	NN	O	B-protein
,	NN	O	O
was	NN	O	O
detected	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
non-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

From	NN	O	O
this	NN	O	O
and	NN	O	O
other	NN	O	O
findings	NN	O	O
the	NN	O	O
picture	NN	O	O
emerges	NN	O	O
that	NN	O	O
there	NN	O	O
are	NN	O	O
at	NN	O	O
least	NN	O	O
three	NN	O	O
categories	NN	O	O
of	NN	O	O
factors	NN	O	O
which	NN	O	O
mediate	NN	O	O
cell-type-specific	NN	O	O
transcription	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
:	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
cell-specific	NN	O	B-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
Oct-2A	NN	O	B-protein
and	NN	O	O
Oct-2B	NN	O	B-protein
that	NN	O	O
are	NN	O	O
not	NN	O	O
expressed	NN	O	O
in	NN	O	O
most	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
:	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
ubiquitous	NN	O	B-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
that	NN	O	O
are	NN	O	O
constitutively	NN	O	O
active	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
are	NN	O	O
sequestered	NN	O	O
in	NN	O	O
an	NN	O	O
inactive	NN	O	O
form	NN	O	O
in	NN	O	O
other	NN	O	O
cells	NN	O	O
;	NN	O	O
(	NN	O	O
c	NN	O	O
)	NN	O	O
ubiquitously	NN	O	B-protein
active	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
exemplified	NN	O	O
by	NN	O	O
the	NN	O	O
one	NN	O	O
binding	NN	O	O
to	NN	O	O
the	NN	O	O
E3	NN	O	B-DNA
sequence	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
factor	NN	O	O
is	NN	O	O
present	NN	O	O
in	NN	O	O
an	NN	O	O
active	NN	O	O
form	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
but	NN	O	O
is	NN	O	O
apparently	NN	O	O
unable	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
Ig	NN	O	I-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
non-B	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
perhaps	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
non-permissive	NN	O	O
chromatin	NN	O	B-DNA
structure	NN	O	O
of	NN	O	O
the	NN	O	O
Ig	NN	O	B-DNA
heavy	NN	O	I-DNA
chain	NN	O	I-DNA
locus	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Endocrine	NN	O	O
status	NN	O	O
changes	NN	O	O
in	NN	O	O
children	NN	O	O
with	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
]	NN	O	O

A	NN	O	O
study	NN	O	O
was	NN	O	O
made	NN	O	O
of	NN	O	O
adrenocortical	NN	O	O
function	NN	O	O
by	NN	O	O
measuring	NN	O	O
blood	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentration	NN	O	O
and	NN	O	O
amount	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
as	NN	O	O
well	NN	O	O
as	NN	O	O
thyroid	NN	O	O
function	NN	O	O
by	NN	O	O
measuring	NN	O	O
blood	NN	O	O
plasma	NN	O	O
triidothyronine	NN	O	O
and	NN	O	O
thyroxine	NN	O	O
concentration	NN	O	O
in	NN	O	O
58	NN	O	O
bronchial	NN	O	O
asthma	NN	O	O
children	NN	O	O
aged	NN	O	O
1	NN	O	O
to	NN	O	O
14	NN	O	O
years	NN	O	O
.	NN	O	O

The	NN	O	O
authors	NN	O	O
revealed	NN	O	O
alterations	NN	O	O
in	NN	O	O
the	NN	O	O
functional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
indicated	NN	O	O
endocrine	NN	O	O
glands	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
intensity	NN	O	O
of	NN	O	O
bronchial	NN	O	O
patency	NN	O	O
disorders	NN	O	O
and	NN	O	O
the	NN	O	O
nature	NN	O	O
of	NN	O	O
the	NN	O	O
therapeutic	NN	O	O
measures	NN	O	O
carried	NN	O	O
out	NN	O	O
.	NN	O	O

-DOCSTART-	O

TAR	NN	O	B-DNA
independent	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
in	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
stimulated	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Multiple	NN	O	B-DNA
regulatory	NN	O	I-DNA
elements	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
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
(	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
)	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
HIV	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Previous	NN	O	O
transfection	NN	O	O
studies	NN	O	O
of	NN	O	O
HIV	NN	O	B-DNA
LTR	NN	O	I-DNA
constructs	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
gene	NN	O	I-DNA
indicated	NN	O	O
that	NN	O	O
multiple	NN	O	O
regulatory	NN	O	B-DNA
regions	NN	O	I-DNA
including	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
,	NN	O	O
SP1	NN	O	B-DNA
,	NN	O	O
TATA	NN	O	B-DNA
and	NN	O	O
TAR	NN	O	B-DNA
regions	NN	O	I-DNA
were	NN	O	O
important	NN	O	O
for	NN	O	O
HIV	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

To	NN	O	O
characterize	NN	O	O
these	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
further	NN	O	O
,	NN	O	O
mutations	NN	O	O
in	NN	O	O
these	NN	O	O
regions	NN	O	O
were	NN	O	O
inserted	NN	O	O
into	NN	O	O
both	NN	O	O
the	NN	O	O
5	NN	O	O
'	NN	O	O
and	NN	O	O
3	NN	O	O
'	NN	O	O
HIV	NN	O	B-DNA
LTRs	NN	O	I-DNA
and	NN	O	O
infectious	NN	O	O
proviral	NN	O	O
constructs	NN	O	O
were	NN	O	O
assembled	NN	O	O
.	NN	O	O

These	NN	O	O
constructs	NN	O	O
were	NN	O	O
transfected	NN	O	O
into	NN	O	O
either	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
both	NN	O	O
the	NN	O	O
presence	NN	O	O
and	NN	O	O
absence	NN	O	O
of	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
which	NN	O	O
have	NN	O	O
previously	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
activate	NN	O	O
HIV	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

Viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
was	NN	O	O
assayed	NN	O	O
by	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
p24	NN	O	B-protein
gag	NN	O	I-protein
protein	NN	O	I-protein
released	NN	O	O
from	NN	O	O
cultures	NN	O	O
transfected	NN	O	O
with	NN	O	O
the	NN	O	O
proviral	NN	O	O
constructs	NN	O	O
.	NN	O	O

Results	NN	O	O
in	NN	O	O
all	NN	O	O
cell	NN	O	O
lines	NN	O	O
indicated	NN	O	O
that	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
SP1	NN	O	B-DNA
,	NN	O	O
TATA	NN	O	B-DNA
and	NN	O	O
the	NN	O	O
TAR	NN	O	O
loop	NN	O	O
and	NN	O	O
stem	NN	O	O
secondary	NN	O	O
structure	NN	O	O
resulted	NN	O	O
in	NN	O	O
marked	NN	O	O
decreases	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
while	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
motif	NN	O	I-DNA
or	NN	O	O
TAR	NN	O	B-DNA
primary	NN	O	I-DNA
sequence	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
only	NN	O	O
slight	NN	O	O
decreases	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
viruses	NN	O	O
containing	NN	O	O
mutations	NN	O	O
in	NN	O	O
either	NN	O	O
the	NN	O	O
TAR	NN	O	B-DNA
loop	NN	O	I-DNA
sequences	NN	O	I-DNA
or	NN	O	O
stem	NN	O	O
secondary	NN	O	O
structure	NN	O	O
which	NN	O	O
were	NN	O	O
very	NN	O	O
defective	NN	O	O
for	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
untreated	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
gave	NN	O	O
nearly	NN	O	O
wild-type	NN	O	O
levels	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
phorbol	NN	O	B-cell_line
ester-treated	NN	O	I-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
phorbol	NN	O	B-cell_line
ester-treated	NN	O	I-cell_line
HeLa	NN	O	I-cell_line
or	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

High	NN	O	O
level	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
TAR	NN	O	B-DNA
mutant	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
phorbol	NN	O	B-cell_line
ester-treated	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
eliminated	NN	O	O
by	NN	O	O
second	NN	O	O
site	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
region	NN	O	I-DNA
or	NN	O	O
by	NN	O	O
disruption	NN	O	O
of	NN	O	O
the	NN	O	O
tat	NN	O	B-DNA
gene	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

A	NN	O	O
case	NN	O	O
of	NN	O	O
hypersensitivity	NN	O	O
to	NN	O	O
thyroid	NN	O	O
hormones	NN	O	O
with	NN	O	O
normally	NN	O	O
functioning	NN	O	O
thyroid	NN	O	O
gland	NN	O	O
and	NN	O	O
increased	NN	O	O
nuclear	NN	O	O
triiodothyronine	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

A	NN	O	O
52-year-old	NN	O	O
male	NN	O	O
presented	NN	O	O
himself	NN	O	O
with	NN	O	O
tachycardia	NN	O	O
crises	NN	O	O
which	NN	O	O
appeared	NN	O	O
first	NN	O	O
during	NN	O	O
childhood	NN	O	O
,	NN	O	O
increased	NN	O	O
in	NN	O	O
frequency	NN	O	O
without	NN	O	O
goiter	NN	O	O
or	NN	O	O
exophthalmos	NN	O	O
.	NN	O	O

Cardiac	NN	O	O
and	NN	O	O
adrenergic	NN	O	O
diseases	NN	O	O
were	NN	O	O
excluded	NN	O	O
.	NN	O	O

The	NN	O	O
thyroid	NN	O	O
function	NN	O	O
was	NN	O	O
normal	NN	O	O
regarding	NN	O	O
T4	NN	O	O
,	NN	O	O
free	NN	O	O
T4	NN	O	O
and	NN	O	O
T3	NN	O	O
,	NN	O	O
TBG	NN	O	O
,	NN	O	O
radioiodine	NN	O	O
uptake	NN	O	O
,	NN	O	O
TSH	NN	O	O
and	NN	O	O
T3	NN	O	O
suppressibility	NN	O	O
;	NN	O	O
however	NN	O	O
the	NN	O	O
TSH	NN	O	O
response	NN	O	O
to	NN	O	O
TRH	NN	O	O
was	NN	O	O
decreased	NN	O	O
.	NN	O	O

The	NN	O	O
lymphocyte	NN	O	B-protein
nuclear	NN	O	I-protein
T3	NN	O	I-protein
receptor	NN	O	I-protein
was	NN	O	O
found	NN	O	O
with	NN	O	O
an	NN	O	O
affinity	NN	O	O
close	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
normal	NN	O	O
volunteers	NN	O	O
(	NN	O	O
Ka	NN	O	O
:	NN	O	O
1.42	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
10	NN	O	O
)	NN	O	O
M-1	NN	O	O
vs	NN	O	O
1.95	NN	O	O
+/-	NN	O	O
0.35	NN	O	O
x	NN	O	O
10	NN	O	O
(	NN	O	O
10	NN	O	O
)	NN	O	O
M-1	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
binding	NN	O	O
capacity	NN	O	O
markedly	NN	O	O
increased	NN	O	O
(	NN	O	O
9.9	NN	O	O
vs	NN	O	O
3.7	NN	O	O
+/-	NN	O	O
0.4	NN	O	O
fmol	NN	O	O
T3/100	NN	O	O
micrograms	NN	O	O
DNA	NN	O	O
)	NN	O	O
.	NN	O	O

Pindolol	NN	O	O
was	NN	O	O
inefficient	NN	O	O
on	NN	O	O
the	NN	O	O
dysrhythmia	NN	O	O
which	NN	O	O
disappeared	NN	O	O
with	NN	O	O
carbimazole	NN	O	O
and	NN	O	O
relapsed	NN	O	O
after	NN	O	O
withdrawal	NN	O	O
of	NN	O	O
the	NN	O	O
antithyroid	NN	O	O
drug	NN	O	O
.	NN	O	O

Under	NN	O	O
carbimazole	NN	O	O
,	NN	O	O
the	NN	O	O
plasma	NN	O	O
T4	NN	O	O
markedly	NN	O	O
decreased	NN	O	O
(	NN	O	O
27.7	NN	O	O
+/-	NN	O	O
3.6	NN	O	O
nmol/l	NN	O	O
)	NN	O	O
but	NN	O	O
the	NN	O	O
patient	NN	O	O
remained	NN	O	O
euthyroid	NN	O	O
.	NN	O	O

The	NN	O	O
clinical	NN	O	O
course	NN	O	O
and	NN	O	O
the	NN	O	O
laboratory	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
tachycardia	NN	O	O
crises	NN	O	O
are	NN	O	O
the	NN	O	O
consequence	NN	O	O
of	NN	O	O
a	NN	O	O
hypersensitivity	NN	O	O
of	NN	O	O
the	NN	O	O
heart	NN	O	O
to	NN	O	O
thyroid	NN	O	O
hormones	NN	O	O
,	NN	O	O
associated	NN	O	O
with	NN	O	O
an	NN	O	O
increased	NN	O	O
number	NN	O	O
of	NN	O	O
T3	NN	O	B-protein
nuclear	NN	O	I-protein
receptor	NN	O	I-protein
sites	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Induction	NN	O	O
of	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
response	NN	O	I-DNA
genes	NN	O	I-DNA
by	NN	O	O
macrophage	NN	O	B-protein
colony-stimulating	NN	O	I-protein
factor	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

A	NN	O	O
group	NN	O	O
of	NN	O	O
coordinately	NN	O	O
induced	NN	O	O
protooncogenes	NN	O	B-DNA
,	NN	O	O
cytoskeletal	NN	O	B-DNA
,	NN	O	I-DNA
and	NN	O	I-DNA
extracellular	NN	O	I-DNA
matrix	NN	O	I-DNA
genes	NN	O	I-DNA
have	NN	O	O
been	NN	O	O
termed	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
their	NN	O	O
induction	NN	O	O
has	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
growth	NN	O	B-protein
factor	NN	O	I-protein
-stimulated	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
the	NN	O	O
induction	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
by	NN	O	O
macrophage-CSF	NN	O	B-protein
(	NN	O	O
M-CSF	NN	O	B-protein
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
that	NN	O	O
do	NN	O	O
not	NN	O	O
proliferate	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
M-CSF	NN	O	B-protein
but	NN	O	O
require	NN	O	O
the	NN	O	O
factor	NN	O	O
for	NN	O	O
optimal	NN	O	O
cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Normal	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
were	NN	O	O
isolated	NN	O	O
,	NN	O	O
carefully	NN	O	O
washed	NN	O	O
,	NN	O	O
and	NN	O	O
incubated	NN	O	O
for	NN	O	O
36	NN	O	O
to	NN	O	O
48	NN	O	O
h	NN	O	O
in	NN	O	O
fetal	NN	O	O
bovine	NN	O	O
serum-containing	NN	O	O
medium	NN	O	O
.	NN	O	O

At	NN	O	O
the	NN	O	O
end	NN	O	O
of	NN	O	O
this	NN	O	O
incubation	NN	O	O
the	NN	O	O
resting	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
M-CSF	NN	O	B-protein
,	NN	O	O
and	NN	O	O
RNA	NN	O	O
was	NN	O	O
isolated	NN	O	O
for	NN	O	O
analysis	NN	O	O
by	NN	O	O
Northern	NN	O	O
blotting	NN	O	O
.	NN	O	O

RNA	NN	O	O
from	NN	O	O
control	NN	O	O
resting	NN	O	B-cell_type
cells	NN	O	I-cell_type
contained	NN	O	O
low	NN	O	O
to	NN	O	O
undetectable	NN	O	O
levels	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
,	NN	O	O
fibronectin	NN	O	B-RNA
receptor	NN	O	I-RNA
,	NN	O	O
and	NN	O	O
actin	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Within	NN	O	O
15	NN	O	O
to	NN	O	O
30	NN	O	O
min	NN	O	O
of	NN	O	O
addition	NN	O	O
of	NN	O	O
M-CSF	NN	O	B-protein
,	NN	O	O
however	NN	O	O
,	NN	O	O
there	NN	O	O
was	NN	O	O
a	NN	O	O
dramatic	NN	O	O
coordinate	NN	O	O
induction	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
.	NN	O	O

The	NN	O	O
c-jun	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
was	NN	O	O
very	NN	O	O
transient	NN	O	O
and	NN	O	O
was	NN	O	O
not	NN	O	O
detectable	NN	O	O
by	NN	O	O
60	NN	O	O
min	NN	O	O
after	NN	O	O
M-CSF	NN	O	B-protein
addition	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
of	NN	O	O
actin	NN	O	B-RNA
and	NN	O	I-RNA
fibronectin	NN	O	I-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
more	NN	O	O
sustained	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
remained	NN	O	O
elevated	NN	O	O
at	NN	O	O
24	NN	O	O
to	NN	O	O
48	NN	O	O
h	NN	O	O
after	NN	O	O
M-CSF	NN	O	B-protein
addition	NN	O	O
.	NN	O	O

We	NN	O	O
also	NN	O	O
observed	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
myelomonocytic	NN	O	B-DNA
specific	NN	O	I-DNA
tyrosine	NN	O	I-DNA
kinase	NN	O	I-DNA
hck	NN	O	I-DNA
gene	NN	O	I-DNA
simultaneously	NN	O	O
with	NN	O	O
the	NN	O	O
other	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

The	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
inhibitor	NN	O	O
cycloheximide	NN	O	O
did	NN	O	O
not	NN	O	O
block	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
any	NN	O	O
of	NN	O	O
these	NN	O	O
genes	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
fact	NN	O	O
,	NN	O	O
super-induced	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
and	NN	O	O
hck	NN	O	B-DNA
.	NN	O	O

Nuclear	NN	O	O
run	NN	O	O
on	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	O
hck	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
actin	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Therefore	NN	O	O
,	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
M-CSF	NN	O	B-protein
induces	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
response	NN	O	I-DNA
genes	NN	O	I-DNA
without	NN	O	O
inducing	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

These	NN	O	O
genes	NN	O	O
may	NN	O	O
then	NN	O	O
play	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
altering	NN	O	O
the	NN	O	O
physiologic	NN	O	O
status	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
CSF	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Inducible	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
kappa	NN	O	B-DNA
B	NN	O	I-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
enhancer	NN	O	I-DNA
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
be	NN	O	O
blocked	NN	O	O
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
in	NN	O	O
a	NN	O	O
signal-dependent	NN	O	O
manner	NN	O	O
.	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
thought	NN	O	O
to	NN	O	O
exert	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
expression	NN	O	O
of	NN	O	O
a	NN	O	O
distinct	NN	O	O
set	NN	O	O
of	NN	O	O
lymphokine	NN	O	B-DNA
genes	NN	O	I-DNA
which	NN	O	O
are	NN	O	O
induced	NN	O	O
upon	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
,	NN	O	O
among	NN	O	O
them	NN	O	O
the	NN	O	O
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
interleukin-2	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
activation	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
is	NN	O	O
partially	NN	O	O
suppressed	NN	O	O
.	NN	O	O

To	NN	O	O
better	NN	O	O
understand	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanisms	NN	O	O
underlying	NN	O	O
suppression	NN	O	O
by	NN	O	O
CsA	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
this	NN	O	O
drug	NN	O	O
on	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
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
that	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
two	NN	O	O
distinct	NN	O	O
mitogen-inducible	NN	O	B-protein
DNA-binding	NN	O	I-protein
complexes	NN	O	I-protein
,	NN	O	O
the	NN	O	O
kappa	NN	O	B-protein
B	NN	O	I-protein
complex	NN	O	I-protein
within	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
NFAT-1	NN	O	B-protein
complex	NN	O	I-protein
within	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
inhibited	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
CsA	NN	O	O
.	NN	O	O

The	NN	O	O
kappa	NN	O	O
B-binding	NN	O	O
activity	NN	O	O
with	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
enhancer	NN	O	I-DNA
is	NN	O	O
inhibited	NN	O	O
only	NN	O	O
if	NN	O	O
it	NN	O	O
is	NN	O	O
activated	NN	O	O
via	NN	O	O
the	NN	O	O
mitogen	NN	O	B-protein
phytohemagglutinin	NN	O	B-protein
whereas	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate-mediated	NN	O	O
activation	NN	O	O
is	NN	O	O
completely	NN	O	O
insensitive	NN	O	O
to	NN	O	O
the	NN	O	O
drug	NN	O	O
.	NN	O	O

This	NN	O	O
suggests	NN	O	O
a	NN	O	O
model	NN	O	O
in	NN	O	O
which	NN	O	O
functionally	NN	O	O
indistinguishable	NN	O	O
kappa	NN	O	B-protein
B	NN	O	I-protein
complexes	NN	O	I-protein
can	NN	O	O
be	NN	O	O
activated	NN	O	O
via	NN	O	O
two	NN	O	O
separate	NN	O	O
pathways	NN	O	O
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
distinguishable	NN	O	O
by	NN	O	O
CsA	NN	O	O
.	NN	O	O

-DOCSTART-	O

Purification	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
a	NN	O	O
T-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
activates	NN	O	O
the	NN	O	O
T-cell	NN	O	B-DNA
receptor	NN	O	I-DNA
C	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
context-dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

The	NN	O	O
differentiation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
into	NN	O	O
functionally	NN	O	O
diverse	NN	O	O
subpopulations	NN	O	O
is	NN	O	O
controlled	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
and	NN	O	O
silencing	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
in	NN	O	O
detail	NN	O	O
about	NN	O	O
the	NN	O	O
proteins	NN	O	O
that	NN	O	O
influence	NN	O	O
this	NN	O	O
developmental	NN	O	O
process	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
purified	NN	O	O
a	NN	O	O
new	NN	O	O
T-cell-specific	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
that	NN	O	O
is	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
genes	NN	O	O
encoding	NN	O	O
a	NN	O	O
major	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
receptor	NN	O	I-protein
(	NN	O	O
TCR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
originally	NN	O	O
identified	NN	O	O
and	NN	O	O
purified	NN	O	O
through	NN	O	O
its	NN	O	O
binding	NN	O	O
sites	NN	O	O
on	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
alpha	NN	O	I-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
to	NN	O	O
promoters	NN	O	B-DNA
for	NN	O	O
several	NN	O	O
genes	NN	O	B-DNA
expressed	NN	O	O
at	NN	O	O
significantly	NN	O	O
earlier	NN	O	O
stages	NN	O	O
of	NN	O	O
T-cell	NN	O	O
development	NN	O	O
than	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
(	NN	O	O
e.g.	NN	O	O
,	NN	O	O
p56lck	NN	O	B-DNA
and	NN	O	O
CD3	NN	O	B-DNA
delta	NN	O	I-DNA
)	NN	O	O
.	NN	O	O

Sequences	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
TCF-1	NN	O	B-DNA
alpha	NN	O	I-DNA
binding	NN	O	I-DNA
motif	NN	O	I-DNA
(	NN	O	O
5'-GGCACCCTTTGA-3	NN	O	O
'	NN	O	O
)	NN	O	O
are	NN	O	O
also	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
TCR	NN	O	I-DNA
delta	NN	O	I-DNA
(	NN	O	O
and	NN	O	O
possibly	NN	O	O
TCR	NN	O	B-DNA
beta	NN	O	I-DNA
)	NN	O	O
enhancers	NN	O	B-DNA
.	NN	O	O

Southwestern	NN	O	O
and	NN	O	O
gel	NN	O	O
renaturation	NN	O	O
experiments	NN	O	O
with	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
purified	NN	O	O
protein	NN	O	O
fractions	NN	O	O
revealed	NN	O	O
that	NN	O	O
TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
activity	NN	O	O
is	NN	O	O
derived	NN	O	O
from	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
57-	NN	O	B-protein
to	NN	O	I-protein
53-kD	NN	O	I-protein
proteins	NN	O	I-protein
that	NN	O	O
are	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_line
and	NN	O	I-cell_line
immature	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
,	NN	O	O
CCRF-CEM	NN	O	B-cell_line
)	NN	O	O
and	NN	O	O
not	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
JY	NN	O	B-cell_line
,	NN	O	O
Namalwa	NN	O	B-cell_line
)	NN	O	O
or	NN	O	O
nonlymphoid	NN	O	B-cell_line
(	NN	O	I-cell_line
HeLa	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

A	NN	O	O
small	NN	O	O
95-bp	NN	O	O
fragment	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
alpha	NN	O	I-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
that	NN	O	O
contains	NN	O	O
the	NN	O	O
TCF-1	NN	O	B-DNA
alpha	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
juxtaposed	NN	O	O
between	NN	O	O
a	NN	O	O
cAMP-response	NN	O	B-DNA
element	NN	O	I-DNA
(	NN	O	O
the	NN	O	O
CRE	NN	O	B-DNA
or	NN	O	O
T	NN	O	B-DNA
alpha	NN	O	I-DNA
1	NN	O	I-DNA
motif	NN	O	I-DNA
)	NN	O	O
and	NN	O	O
the	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
a	NN	O	O
distinct	NN	O	O
lymphoid-specific	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
TCF-2	NN	O	B-protein
alpha	NN	O	I-protein
)	NN	O	O
behaved	NN	O	O
as	NN	O	O
a	NN	O	O
potent	NN	O	O
T-cell-specific	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Tandem	NN	O	O
copies	NN	O	O
of	NN	O	O
this	NN	O	O
enhancer	NN	O	O
functioned	NN	O	O
synergistically	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_line
(	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
)	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
as	NN	O	O
resting	NN	O	O
and	NN	O	O
activated	NN	O	O
immature	NN	O	B-cell_line
(	NN	O	I-cell_line
CCRF-CEM	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

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
TCF-1	NN	O	B-DNA
alpha	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
diminished	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
and	NN	O	O
disrupted	NN	O	O
the	NN	O	O
synergism	NN	O	O
observed	NN	O	O
in	NN	O	O
vivo	NN	O	O
between	NN	O	O
tandem	NN	O	B-DNA
enhancer	NN	O	I-DNA
repeats	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
TCF-1	NN	O	B-DNA
alpha	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
also	NN	O	O
required	NN	O	O
for	NN	O	O
TCR	NN	O	B-protein
alpha	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
in	NN	O	O
transcriptionally	NN	O	O
active	NN	O	O
extracts	NN	O	O
from	NN	O	O
Jurkat	NN	O	B-cell_line
but	NN	O	O
not	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
confirming	NN	O	O
that	NN	O	O
TCF-1	NN	O	B-protein
alpha	NN	O	I-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

Curiously	NN	O	O
,	NN	O	O
the	NN	O	O
TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
binding	NN	O	O
element	NN	O	O
was	NN	O	O
inactive	NN	O	O
in	NN	O	O
vivo	NN	O	O
when	NN	O	O
removed	NN	O	O
from	NN	O	O
its	NN	O	O
neighboring	NN	O	O
elements	NN	O	O
on	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
alpha	NN	O	I-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
positioned	NN	O	O
in	NN	O	O
one	NN	O	O
or	NN	O	O
more	NN	O	O
copies	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
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
TCF-1	NN	O	B-protein
alpha	NN	O	I-protein
appears	NN	O	O
to	NN	O	O
depend	NN	O	O
on	NN	O	O
the	NN	O	O
TCF-2	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
T	NN	O	B-protein
alpha	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
CREB	NN	O	B-protein
)	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
its	NN	O	O
binding	NN	O	O
site	NN	O	O
within	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

-DOCSTART-	O

Tandem	NN	O	B-DNA
AP-1-binding	NN	O	I-DNA
sites	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
beta-globin	NN	O	I-DNA
dominant	NN	O	I-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
function	NN	O	O
as	NN	O	O
an	NN	O	O
inducible	NN	O	B-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

A	NN	O	O
powerful	NN	O	O
enhancer	NN	O	O
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
an	NN	O	O
18-bp	NN	O	B-DNA
DNA	NN	O	I-DNA
segment	NN	O	I-DNA
located	NN	O	O
11	NN	O	O
kb	NN	O	O
5	NN	O	O
'	NN	O	O
to	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
epsilon-globin	NN	O	I-DNA
gene	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
dominant	NN	O	B-DNA
control	NN	O	I-DNA
or	NN	O	O
locus-activating	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
enhancer	NN	O	B-DNA
is	NN	O	O
inducible	NN	O	O
in	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
increasing	NN	O	O
linked	NN	O	O
gamma-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
/luciferase	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
to	NN	O	O
170-fold	NN	O	O
over	NN	O	O
an	NN	O	O
enhancerless	NN	O	B-DNA
construct	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
enhancer	NN	O	B-DNA
consists	NN	O	O
of	NN	O	O
tandem	NN	O	B-DNA
AP-1-binding	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
phased	NN	O	O
10	NN	O	O
bp	NN	O	O
apart	NN	O	O
,	NN	O	O
which	NN	O	O
are	NN	O	O
both	NN	O	O
required	NN	O	O
for	NN	O	O
full	NN	O	O
activity	NN	O	O
.	NN	O	O

DNA-protein	NN	O	O
binding	NN	O	O
assays	NN	O	O
with	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
induced	NN	O	O
cells	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
high	NN	O	B-protein
molecular	NN	O	I-protein
weight	NN	O	I-protein
complex	NN	O	I-protein
on	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
.	NN	O	O

The	NN	O	O
formation	NN	O	O
of	NN	O	O
this	NN	O	O
complex	NN	O	O
also	NN	O	O
requires	NN	O	O
both	NN	O	O
AP-1	NN	O	B-DNA
sites	NN	O	I-DNA
and	NN	O	O
correlates	NN	O	O
with	NN	O	O
maximal	NN	O	O
enhancer	NN	O	B-DNA
activity	NN	O	O
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
may	NN	O	O
have	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
increase	NN	O	O
in	NN	O	O
globin	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
that	NN	O	O
characterizes	NN	O	O
erythroid	NN	O	O
maturation	NN	O	O
.	NN	O	O

Enhancer	NN	O	O
activity	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
complex	NN	O	O
of	NN	O	O
proteins	NN	O	O
from	NN	O	O
the	NN	O	O
jun	NN	O	B-DNA
and	NN	O	I-DNA
fos	NN	O	I-DNA
families	NN	O	I-DNA
to	NN	O	O
tandem	NN	O	B-DNA
AP-1	NN	O	I-DNA
consensus	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
factor	NN	O	I-protein
that	NN	O	O
interacts	NN	O	O
with	NN	O	O
an	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
promoter	NN	O	I-DNA
and	NN	O	O
stimulates	NN	O	O
transcription	NN	O	O
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
the	NN	O	O
lymphoid	NN	O	B-protein
cell-specific	NN	O	I-protein
factor	NN	O	I-protein
OTF2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
MOPC	NN	O	B-DNA
141	NN	O	I-DNA
immunoglobulin	NN	O	I-DNA
heavy-chain	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
studied	NN	O	O
by	NN	O	O
using	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
.	NN	O	O

B-cell-specific	NN	O	O
transcription	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
element	NN	O	I-DNA
5'-ATGCAAAG-3	NN	O	O
'	NN	O	O
,	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
region	NN	O	I-DNA
of	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
and	NN	O	O
in	NN	O	O
the	NN	O	O
promoters	NN	O	B-DNA
of	NN	O	O
all	NN	O	O
other	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-	NN	O	I-DNA
and	NN	O	I-DNA
light-chain	NN	O	I-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
interaction	NN	O	O
of	NN	O	O
purified	NN	O	O
octamer	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
1	NN	O	O
and	NN	O	O
2	NN	O	O
(	NN	O	O
OTF1	NN	O	B-protein
and	NN	O	O
OTF2	NN	O	B-protein
)	NN	O	O
with	NN	O	O
the	NN	O	O
MOPC	NN	O	B-DNA
141	NN	O	I-DNA
promoter	NN	O	I-DNA
was	NN	O	O
studied	NN	O	O
by	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
DNase	NN	O	B-protein
I	NN	O	I-protein
footprinting	NN	O	O
.	NN	O	O

Purified	NN	O	O
OTF1	NN	O	B-protein
from	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
OTF1	NN	O	B-protein
and	NN	O	O
OTF2	NN	O	B-protein
from	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
bound	NN	O	O
to	NN	O	O
identical	NN	O	O
sequences	NN	O	O
within	NN	O	O
the	NN	O	O
heavy-chain	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
OTF	NN	O	O
interactions	NN	O	O
we	NN	O	O
observed	NN	O	O
extended	NN	O	O
over	NN	O	O
the	NN	O	O
heptamer	NN	O	B-DNA
element	NN	O	I-DNA
5'-CTCAGGA-3	NN	O	O
'	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
seems	NN	O	O
likely	NN	O	O
that	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
purified	NN	O	B-protein
factors	NN	O	I-protein
involves	NN	O	O
cooperation	NN	O	O
between	NN	O	O
octamer	NN	O	B-DNA
and	NN	O	I-DNA
heptamer	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
this	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
these	NN	O	O
elements	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
second	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
the	NN	O	O
N	NN	O	B-DNA
element	NN	O	I-DNA
with	NN	O	O
the	NN	O	O
sequence	NN	O	O
5'-GGAACCTCCCCC-3'	NN	O	O
.	NN	O	O

The	NN	O	O
N	NN	O	B-DNA
element	NN	O	I-DNA
could	NN	O	O
independently	NN	O	O
mediate	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
transcription	NN	O	O
in	NN	O	O
both	NN	O	O
B-cell	NN	O	O
and	NN	O	O
HeLa-cell	NN	O	O
extracts	NN	O	O
,	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
conjunction	NN	O	O
with	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
it	NN	O	O
can	NN	O	O
promote	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
transcription	NN	O	O
in	NN	O	O
B-cell	NN	O	O
extracts	NN	O	O
.	NN	O	O

The	NN	O	O
N	NN	O	B-DNA
element	NN	O	I-DNA
bound	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
NTF	NN	O	B-protein
,	NN	O	O
that	NN	O	O
is	NN	O	O
ubiquitous	NN	O	O
in	NN	O	O
cell-type	NN	O	O
distribution	NN	O	O
,	NN	O	O
and	NN	O	O
NTF	NN	O	B-protein
was	NN	O	O
distinct	NN	O	O
from	NN	O	O
any	NN	O	O
of	NN	O	O
the	NN	O	O
previously	NN	O	B-protein
described	NN	O	I-protein
proteins	NN	O	I-protein
that	NN	O	O
bind	NN	O	O
to	NN	O	O
similar	NN	O	O
sequences	NN	O	O
.	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
propose	NN	O	O
that	NN	O	O
NTF	NN	O	B-protein
and	NN	O	O
OTF2	NN	O	B-protein
interactions	NN	O	O
(	NN	O	O
both	NN	O	O
with	NN	O	O
their	NN	O	O
cognate	NN	O	B-DNA
DNA	NN	O	I-DNA
elements	NN	O	I-DNA
and	NN	O	O
possibly	NN	O	O
at	NN	O	O
the	NN	O	O
protein-protein	NN	O	O
level	NN	O	O
)	NN	O	O
may	NN	O	O
be	NN	O	O
critical	NN	O	O
to	NN	O	O
B-cell-specific	NN	O	O
expression	NN	O	O
and	NN	O	O
that	NN	O	O
these	NN	O	O
interactions	NN	O	O
provide	NN	O	O
additional	NN	O	O
pathways	NN	O	O
for	NN	O	O
regulating	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-kappa	NN	O	B-protein
B	NN	O	I-protein
as	NN	O	O
inducible	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
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
.	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
the	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
induced	NN	O	O
upon	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
phytohemagglutinin	NN	O	B-protein
and	NN	O	O
active	NN	O	O
phorbolester	NN	O	O
and	NN	O	O
upon	NN	O	O
expression	NN	O	O
of	NN	O	O
tax1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
transactivating	NN	O	B-protein
protein	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

The	NN	O	O
same	NN	O	O
agents	NN	O	O
induce	NN	O	O
transcription	NN	O	O
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
and	NN	O	I-DNA
interleukin-2	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
depending	NN	O	O
on	NN	O	O
promoter	NN	O	B-DNA
elements	NN	O	I-DNA
that	NN	O	O
bind	NN	O	O
the	NN	O	O
inducible	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
or	NN	O	O
an	NN	O	O
NF-kappa	NN	O	B-protein
B-like	NN	O	I-protein
factor	NN	O	I-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
tested	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
also	NN	O	O
regulated	NN	O	O
by	NN	O	O
a	NN	O	O
cognate	NN	O	O
motif	NN	O	O
for	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

A	NN	O	O
recent	NN	O	O
functional	NN	O	O
analysis	NN	O	O
by	NN	O	O
Miyatake	NN	O	O
et	NN	O	O
al.	NN	O	O
(	NN	O	O
S.	NN	O	O
Miyatake	NN	O	O
,	NN	O	O
M.	NN	O	O
Seiki	NN	O	O
,	NN	O	O
M.	NN	O	O
Yoshida	NN	O	O
,	NN	O	O
and	NN	O	O
K.	NN	O	O
Arai	NN	O	O
,	NN	O	O
Mol.	NN	O	O
Cell.	NN	O	O
Biol.	NN	O	O
8	NN	O	O
:	NN	O	O
5581-5587	NN	O	O
,	NN	O	O
1988	NN	O	O
)	NN	O	O
described	NN	O	O
a	NN	O	O
short	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
conferred	NN	O	O
strong	NN	O	O
inducibility	NN	O	O
by	NN	O	O
T-cell-activating	NN	O	O
signals	NN	O	O
and	NN	O	O
tax1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
no	NN	O	O
NF-kappa	NN	O	B-DNA
B-binding	NN	O	I-DNA
motifs	NN	O	I-DNA
were	NN	O	O
identified	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
showed	NN	O	O
binding	NN	O	O
of	NN	O	O
purified	NN	O	O
human	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
of	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activated	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
to	NN	O	O
an	NN	O	O
oligonucleotide	NN	O	O
comprising	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
responsible	NN	O	O
for	NN	O	O
mediating	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
T-cell-activating	NN	O	O
signals	NN	O	O
and	NN	O	O
tax1	NN	O	B-protein
.	NN	O	O

As	NN	O	O
shown	NN	O	O
by	NN	O	O
a	NN	O	O
methylation	NN	O	O
interference	NN	O	O
analysis	NN	O	O
and	NN	O	O
oligonucleotide	NN	O	O
competition	NN	O	O
experiments	NN	O	O
,	NN	O	O
purified	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binds	NN	O	O
at	NN	O	O
positions	NN	O	B-DNA
-82	NN	O	I-DNA
to	NN	O	I-DNA
-91	NN	O	I-DNA
(	NN	O	O
GGGAACTACC	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
sequence	NN	O	O
with	NN	O	O
an	NN	O	O
affinity	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
with	NN	O	O
which	NN	O	O
it	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
biologically	NN	O	B-DNA
functional	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
motif	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
beta	NN	O	B-DNA
interferon	NN	O	I-DNA
promoter	NN	O	I-DNA
(	NN	O	O
GGGAAATTCC	NN	O	O
)	NN	O	O
.	NN	O	O

Two	NN	O	O
kappa	NN	O	B-DNA
B-like	NN	O	I-DNA
motifs	NN	O	I-DNA
at	NN	O	O
positions	NN	O	B-DNA
-98	NN	O	I-DNA
to	NN	O	I-DNA
-108	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
were	NN	O	O
also	NN	O	O
recognized	NN	O	O
but	NN	O	O
with	NN	O	O
much	NN	O	O
lower	NN	O	O
affinities	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
provide	NN	O	O
strong	NN	O	O
evidence	NN	O	O
that	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
gene	NN	O	I-DNA
following	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
is	NN	O	O
controlled	NN	O	O
by	NN	O	O
binding	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
to	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
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
mitogenic	NN	O	O
agents	NN	O	O
upon	NN	O	O
glucocorticoid	NN	O	O
action	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
tonsillar	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
treatment	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
tonsillar	NN	O	I-cell_type
T-lymphocytes	NN	O	I-cell_type
with	NN	O	O
4-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
resulted	NN	O	O
in	NN	O	O
about	NN	O	O
two	NN	O	O
fold	NN	O	O
increase	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
number	NN	O	O
,	NN	O	O
without	NN	O	O
any	NN	O	O
significant	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
receptor	NN	O	O
affinity	NN	O	O
.	NN	O	O

This	NN	O	O
increase	NN	O	O
disappeared	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

Alone	NN	O	O
,	NN	O	O
PMA	NN	O	O
and	NN	O	O
calcium	NN	O	O
ionophore	NN	O	O
A23187	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
,	NN	O	O
but	NN	O	O
together	NN	O	O
stimulated	NN	O	O
,	NN	O	O
like	NN	O	O
phytohaemagglutinin	NN	O	B-protein
(	NN	O	O
PHA	NN	O	B-protein
)	NN	O	O
,	NN	O	O
leucine	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
particular	NN	O	O
,	NN	O	O
thymidine	NN	O	O
incorporation	NN	O	O
.	NN	O	O

PMA	NN	O	O
enhanced	NN	O	O
slightly	NN	O	O
the	NN	O	O
stimulatory	NN	O	O
effect	NN	O	O
of	NN	O	O
PHA	NN	O	B-protein
.	NN	O	O

Alone	NN	O	O
,	NN	O	O
these	NN	O	O
agents	NN	O	O
failed	NN	O	O
to	NN	O	O
alter	NN	O	O
the	NN	O	O
suppressive	NN	O	O
effect	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
on	NN	O	O
thymidine	NN	O	O
and	NN	O	O
leucine	NN	O	O
incorporation	NN	O	O
;	NN	O	O
however	NN	O	O
,	NN	O	O
PMA-A23187	NN	O	O
and	NN	O	O
PMA	NN	O	O
-PHA	NN	O	B-protein
combinations	NN	O	O
appeared	NN	O	O
to	NN	O	O
antagonize	NN	O	O
the	NN	O	O
suppression	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
.	NN	O	O

-DOCSTART-	O

To	NN	O	O
be	NN	O	O
or	NN	O	O
not	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
responder	NN	O	O
in	NN	O	O
T-cell	NN	O	O
responses	NN	O	O
:	NN	O	O
ubiquitous	NN	O	O
oligopeptides	NN	O	O
in	NN	O	O
all	NN	O	O
proteins	NN	O	O
.	NN	O	O

Amino	NN	O	O
acid	NN	O	O
sequences	NN	O	O
of	NN	O	O
all	NN	O	O
proteins	NN	O	O
are	NN	O	O
essays	NN	O	O
written	NN	O	O
in	NN	O	O
the	NN	O	O
same	NN	O	O
language	NN	O	O
.	NN	O	O

Accordingly	NN	O	O
,	NN	O	O
the	NN	O	O
same	NN	O	O
set	NN	O	O
of	NN	O	O
words	NN	O	O
and	NN	O	O
phrases	NN	O	O
(	NN	O	O
oligopeptides	NN	O	O
)	NN	O	O
appear	NN	O	O
in	NN	O	O
totally	NN	O	O
unrelated	NN	O	O
proteins	NN	O	O
.	NN	O	O

The	NN	O	O
reason	NN	O	O
that	NN	O	O
only	NN	O	O
certain	NN	O	O
individuals	NN	O	O
of	NN	O	O
particular	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
haplotypes	NN	O	I-protein
can	NN	O	O
mount	NN	O	O
T-cell	NN	O	O
responses	NN	O	O
against	NN	O	O
a	NN	O	O
given	NN	O	O
antigen	NN	O	O
of	NN	O	O
pathogens	NN	O	O
is	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
T-cell	NN	O	B-protein
receptors	NN	O	I-protein
are	NN	O	O
designed	NN	O	O
to	NN	O	O
recognize	NN	O	O
18-20	NN	O	O
residue-long	NN	O	O
peptide	NN	O	O
fragments	NN	O	O
sandwiched	NN	O	O
between	NN	O	O
two	NN	O	O
alpha-helices	NN	O	B-protein
of	NN	O	O
class	NN	O	B-protein
I	NN	O	I-protein
or	NN	O	O
class	NN	O	O
II	NN	O	O
MHC	NN	O	O
molecules	NN	O	O
.	NN	O	O

At	NN	O	O
this	NN	O	O
range	NN	O	O
of	NN	O	O
peptide	NN	O	O
lengths	NN	O	O
,	NN	O	O
most	NN	O	O
would	NN	O	O
appear	NN	O	O
as	NN	O	O
self	NN	O	O
,	NN	O	O
while	NN	O	O
nonselfness	NN	O	O
of	NN	O	O
the	NN	O	O
remainders	NN	O	O
are	NN	O	O
destined	NN	O	O
to	NN	O	O
be	NN	O	O
quite	NN	O	O
ambiguous	NN	O	O
,	NN	O	O
hence	NN	O	O
creating	NN	O	O
responders	NN	O	O
and	NN	O	O
nonresponders	NN	O	O
.	NN	O	O

-DOCSTART-	O

Two	NN	O	O
distinct	NN	O	O
signal	NN	O	O
transmission	NN	O	O
pathways	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
are	NN	O	O
inhibited	NN	O	O
by	NN	O	O
complexes	NN	O	O
formed	NN	O	O
between	NN	O	O
an	NN	O	O
immunophilin	NN	O	B-protein
and	NN	O	O
either	NN	O	O
FK506	NN	O	O
or	NN	O	O
rapamycin	NN	O	O
.	NN	O	O

Proliferation	NN	O	O
and	NN	O	O
immunologic	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
initiated	NN	O	O
by	NN	O	O
signals	NN	O	O
from	NN	O	O
the	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
that	NN	O	O
are	NN	O	O
inhibited	NN	O	O
by	NN	O	O
the	NN	O	O
immunosuppressant	NN	O	O
FK506	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
its	NN	O	O
structural	NN	O	O
analog	NN	O	O
,	NN	O	O
rapamycin	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
interleukin	NN	O	B-protein
2	NN	O	I-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
-induced	NN	O	O
signals	NN	O	O
are	NN	O	O
blocked	NN	O	O
by	NN	O	O
rapamycin	NN	O	O
but	NN	O	O
not	NN	O	O
by	NN	O	O
FK506	NN	O	O
.	NN	O	O

Remarkably	NN	O	O
,	NN	O	O
these	NN	O	O
two	NN	O	O
drugs	NN	O	O
inhibit	NN	O	O
each	NN	O	O
other	NN	O	O
's	NN	O	O
actions	NN	O	O
,	NN	O	O
raising	NN	O	O
the	NN	O	O
possibility	NN	O	O
that	NN	O	O
both	NN	O	O
act	NN	O	O
by	NN	O	O
means	NN	O	O
of	NN	O	O
a	NN	O	O
common	NN	O	O
immunophilin	NN	O	B-protein
(	NN	O	O
immunosuppressant	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
the	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
rapamycin	NN	O	O
to	NN	O	O
the	NN	O	O
FK506	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
FKBP	NN	O	B-protein
(	NN	O	O
Kd	NN	O	O
=	NN	O	O
0.2	NN	O	O
nM	NN	O	O
)	NN	O	O
is	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
of	NN	O	O
FK506	NN	O	O
to	NN	O	O
FKBP	NN	O	B-protein
(	NN	O	O
Kd	NN	O	O
=	NN	O	O
0.4	NN	O	O
nM	NN	O	O
)	NN	O	O
and	NN	O	O
to	NN	O	O
their	NN	O	O
effective	NN	O	O
biologic	NN	O	O
inhibitory	NN	O	O
concentrations	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
an	NN	O	O
excess	NN	O	O
of	NN	O	O
rapamycin	NN	O	O
is	NN	O	O
needed	NN	O	O
to	NN	O	O
revert	NN	O	O
FK506-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
,	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
and	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
a	NN	O	O
T-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
necessary	NN	O	O
for	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
activation	NN	O	O
.	NN	O	O

Similarly	NN	O	O
,	NN	O	O
an	NN	O	O
excess	NN	O	O
of	NN	O	O
FK506	NN	O	O
is	NN	O	O
needed	NN	O	O
to	NN	O	O
revert	NN	O	O
rapamycin-mediated	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
.	NN	O	O

The	NN	O	O
drug	NN	O	O
concentrations	NN	O	O
required	NN	O	O
for	NN	O	O
antagonism	NN	O	O
may	NN	O	O
be	NN	O	O
explained	NN	O	O
by	NN	O	O
the	NN	O	O
relative	NN	O	O
affinity	NN	O	O
of	NN	O	O
the	NN	O	O
drugs	NN	O	O
to	NN	O	O
,	NN	O	O
and	NN	O	O
by	NN	O	O
the	NN	O	O
abundance	NN	O	O
of	NN	O	O
,	NN	O	O
the	NN	O	O
immunophilin	NN	O	B-protein
FKBP	NN	O	B-protein
.	NN	O	O

FKBP	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
catalyze	NN	O	O
the	NN	O	O
interconversion	NN	O	O
of	NN	O	O
the	NN	O	O
cis-	NN	O	O
and	NN	O	O
trans-rotamers	NN	O	O
of	NN	O	O
the	NN	O	O
peptidyl-prolyl	NN	O	B-protein
amide	NN	O	I-protein
bond	NN	O	I-protein
of	NN	O	O
peptide	NN	O	O
substrates	NN	O	O
;	NN	O	O
here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
rapamycin	NN	O	O
,	NN	O	O
like	NN	O	O
FK506	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
potent	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
the	NN	O	O
rotamase	NN	O	O
activity	NN	O	O
of	NN	O	O
FKBP	NN	O	B-protein
(	NN	O	O
Ki	NN	O	O
=	NN	O	O
0.2	NN	O	O
nM	NN	O	O
)	NN	O	O
.	NN	O	O

Neither	NN	O	O
FKBP	NN	O	B-protein
binding	NN	O	O
nor	NN	O	O
inhibition	NN	O	O
of	NN	O	O
rotamase	NN	O	O
activity	NN	O	O
of	NN	O	O
FKBP	NN	O	B-protein
alone	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
explain	NN	O	O
the	NN	O	O
biologic	NN	O	O
actions	NN	O	O
of	NN	O	O
these	NN	O	O
drugs	NN	O	O
.	NN	O	O

Rather	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
immunophilin	NN	O	B-protein
bound	NN	O	O
to	NN	O	O
FK506	NN	O	O
interferes	NN	O	O
with	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
-induced	NN	O	O
signals	NN	O	O
,	NN	O	O
while	NN	O	O
rapamycin	NN	O	O
bound	NN	O	O
to	NN	O	O
the	NN	O	O
immunophilin	NN	O	B-protein
interferes	NN	O	O
with	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
signals	NN	O	O
.	NN	O	O

-DOCSTART-	O

Adherence-dependent	NN	O	O
increase	NN	O	O
in	NN	O	O
human	NN	O	B-RNA
monocyte	NN	O	I-RNA
PDGF	NN	O	I-RNA
(	NN	O	I-RNA
B	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
associated	NN	O	O
with	NN	O	O
increases	NN	O	O
in	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
EGR2	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Adherence	NN	O	O
is	NN	O	O
an	NN	O	O
important	NN	O	O
initial	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
transition	NN	O	O
of	NN	O	O
a	NN	O	O
circulating	NN	O	B-cell_type
monocyte	NN	O	I-cell_type
to	NN	O	O
a	NN	O	O
tissue	NN	O	B-cell_type
macrophage	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
differentiation	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
augmented	NN	O	O
capacity	NN	O	O
to	NN	O	O
generate	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
adherence	NN	O	O
itself	NN	O	O
might	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
trigger	NN	O	O
for	NN	O	O
a	NN	O	O
sequence	NN	O	O
of	NN	O	O
gene	NN	O	O
activation	NN	O	O
culminating	NN	O	O
in	NN	O	O
cells	NN	O	O
with	NN	O	O
increased	NN	O	O
mRNA	NN	O	B-RNA
encoding	NN	O	O
profibrotic	NN	O	O
growth	NN	O	B-protein
factors	NN	O	I-protein
such	NN	O	O
as	NN	O	O
platelet-derived	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
B	NN	O	I-protein
subunit	NN	O	I-protein
(	NN	O	O
PDGF	NN	O	B-protein
[	NN	O	I-protein
B	NN	O	I-protein
]	NN	O	I-protein
)	NN	O	O
and	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

After	NN	O	O
in	NN	O	O
vitro	NN	O	O
adherence	NN	O	O
,	NN	O	O
human	NN	O	O
monocytes	NN	O	O
had	NN	O	O
a	NN	O	O
biphasic	NN	O	O
increase	NN	O	O
in	NN	O	O
PDGF	NN	O	B-RNA
(	NN	O	I-RNA
B	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
with	NN	O	O
peaks	NN	O	O
at	NN	O	O
6	NN	O	O
h	NN	O	O
and	NN	O	O
13	NN	O	O
d	NN	O	O
.	NN	O	O

No	NN	O	O
increase	NN	O	O
in	NN	O	O
TGF-beta	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
observed	NN	O	O
.	NN	O	O

The	NN	O	O
6-h	NN	O	O
increase	NN	O	O
in	NN	O	O
PDGF	NN	O	B-RNA
(	NN	O	I-RNA
B	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
adherence	NN	O	O
dependent	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
addition	NN	O	O
,	NN	O	O
was	NN	O	O
abrogated	NN	O	O
when	NN	O	O
the	NN	O	O
cytoskeletal	NN	O	O
integrity	NN	O	O
was	NN	O	O
compromised	NN	O	O
by	NN	O	O
cytochalasin	NN	O	O
D	NN	O	O
.	NN	O	O

The	NN	O	O
6-h	NN	O	O
increase	NN	O	O
in	NN	O	O
PDGF	NN	O	B-RNA
(	NN	O	I-RNA
B	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
unaltered	NN	O	O
by	NN	O	O
adherence	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
monocyte	NN	O	O
stimulus	NN	O	O
lipopolysaccharide	NN	O	O
.	NN	O	O

Adherence	NN	O	O
to	NN	O	O
either	NN	O	O
fibronectin	NN	O	O
or	NN	O	O
collagen-coated	NN	O	O
plastic	NN	O	O
had	NN	O	O
little	NN	O	O
consistent	NN	O	O
effect	NN	O	O
on	NN	O	O
PDGF	NN	O	B-RNA
(	NN	O	I-RNA
B	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
.	NN	O	O

The	NN	O	O
increased	NN	O	O
PDGF	NN	O	B-RNA
(	NN	O	I-RNA
B	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
observed	NN	O	O
in	NN	O	O
adherent	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
increases	NN	O	O
in	NN	O	O
mRNAs	NN	O	B-RNA
of	NN	O	O
the	NN	O	O
early	NN	O	B-DNA
growth	NN	O	I-DNA
response	NN	O	I-DNA
genes	NN	O	I-DNA
c-fos	NN	O	B-DNA
(	NN	O	O
maximal	NN	O	O
at	NN	O	O
20	NN	O	O
min	NN	O	O
)	NN	O	O
,	NN	O	O
c-jun	NN	O	O
,	NN	O	O
and	NN	O	O
EGR2	NN	O	O
(	NN	O	O
maximal	NN	O	O
at	NN	O	O
6-24	NN	O	O
h	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
increase	NN	O	O
in	NN	O	O
c-jun	NN	O	O
and	NN	O	O
EGR2	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
c-fos	NN	O	B-DNA
,	NN	O	O
mRNA	NN	O	O
was	NN	O	O
also	NN	O	O
abrogated	NN	O	O
by	NN	O	O
cytochalasin	NN	O	O
D	NN	O	O
.	NN	O	O

These	NN	O	O
observations	NN	O	O
suggest	NN	O	O
that	NN	O	O
adherence	NN	O	O
results	NN	O	O
in	NN	O	O
increases	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
EGR2	NN	O	I-RNA
,	NN	O	I-RNA
and	NN	O	I-RNA
PDGF	NN	O	I-RNA
(	NN	O	I-RNA
B	NN	O	I-RNA
)	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
increases	NN	O	O
in	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	O
EGR2	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
PDGF	NN	O	B-protein
(	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	I-protein
may	NN	O	O
depend	NN	O	O
on	NN	O	O
cytoskeletal	NN	O	O
rearrangement	NN	O	O
.	NN	O	O

Modulation	NN	O	O
of	NN	O	O
these	NN	O	O
events	NN	O	O
at	NN	O	O
the	NN	O	O
time	NN	O	O
of	NN	O	O
adherence	NN	O	O
offers	NN	O	O
a	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
differential	NN	O	O
priming	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
may	NN	O	O
be	NN	O	O
accomplished	NN	O	O
.	NN	O	O

-DOCSTART-	O

Single	NN	O	O
cell	NN	O	O
assay	NN	O	O
of	NN	O	O
a	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
reveals	NN	O	O
a	NN	O	O
threshold	NN	O	O
in	NN	O	O
transcription	NN	O	O
activated	NN	O	O
by	NN	O	O
signals	NN	O	O
emanating	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
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
through	NN	O	O
their	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
appearance	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
NF-AT	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
regulating	NN	O	O
genes	NN	O	O
required	NN	O	O
for	NN	O	O
immunologic	NN	O	O
activation	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
single	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
in	NN	O	O
individual	NN	O	B-cell_type
viable	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
we	NN	O	O
have	NN	O	O
applied	NN	O	O
an	NN	O	O
assay	NN	O	O
that	NN	O	O
uses	NN	O	O
the	NN	O	O
fluorescence-activated	NN	O	O
cell	NN	O	O
sorter	NN	O	O
to	NN	O	O
quantitate	NN	O	O
beta-galactosidase	NN	O	B-protein
(	NN	O	O
beta-gal	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
transcriptional	NN	O	O
activity	NN	O	O
among	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
undergoing	NN	O	O
activation	NN	O	O
by	NN	O	O
using	NN	O	O
a	NN	O	O
construct	NN	O	O
in	NN	O	O
which	NN	O	O
three	NN	O	O
tandem	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
NF-AT-binding	NN	O	B-DNA
site	NN	O	I-DNA
directs	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
lacZ	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Unexpectedly	NN	O	O
,	NN	O	O
stimulation	NN	O	O
of	NN	O	O
cloned	NN	O	B-cell_line
stably	NN	O	I-cell_line
transfected	NN	O	I-cell_line
Jurkat	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
leads	NN	O	O
to	NN	O	O
a	NN	O	O
bimodal	NN	O	O
pattern	NN	O	O
of	NN	O	O
beta-gal	NN	O	O
expression	NN	O	O
in	NN	O	O
which	NN	O	O
some	NN	O	O
cells	NN	O	O
express	NN	O	O
no	NN	O	O
beta-gal	NN	O	B-protein
and	NN	O	O
others	NN	O	O
express	NN	O	O
high	NN	O	O
levels	NN	O	O
.	NN	O	O

This	NN	O	O
expression	NN	O	O
pattern	NN	O	O
can	NN	O	O
not	NN	O	O
be	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
cell-cycle	NN	O	O
position	NN	O	O
or	NN	O	O
heritable	NN	O	O
variation	NN	O	O
.	NN	O	O

Further	NN	O	O
results	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
beta-gal	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
correlated	NN	O	O
with	NN	O	O
NF-AT	NN	O	B-protein
-binding	NN	O	O
activity	NN	O	O
,	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
must	NN	O	O
exceed	NN	O	O
a	NN	O	O
critical	NN	O	O
threshold	NN	O	O
before	NN	O	O
transcription	NN	O	O
initiates	NN	O	O
.	NN	O	O

This	NN	O	O
threshold	NN	O	O
likely	NN	O	O
reflects	NN	O	O
the	NN	O	O
NF-AT	NN	O	B-protein
concentration-dependent	NN	O	O
assembly	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
complexes	NN	O	I-protein
at	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
.	NN	O	O

Similar	NN	O	O
constructs	NN	O	O
controlled	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
or	NN	O	O
the	NN	O	O
entire	NN	O	O
interleukin-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
show	NN	O	O
bimodal	NN	O	O
expression	NN	O	O
patterns	NN	O	O
during	NN	O	O
induction	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
thresholds	NN	O	O
set	NN	O	O
by	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	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
common	NN	O	O
property	NN	O	O
of	NN	O	O
inducible	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	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
BMRF1	NN	O	I-DNA
promoter	NN	O	I-DNA
for	NN	O	O
early	NN	O	B-protein
antigen	NN	O	I-protein
(	NN	O	O
EA-D	NN	O	B-protein
)	NN	O	O
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
transactivators	NN	O	I-protein
,	NN	O	O
BRLF1	NN	O	B-protein
and	NN	O	O
BZLF1	NN	O	B-protein
,	NN	O	O
in	NN	O	O
a	NN	O	O
cell-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

The	NN	O	O
Epstein-Barr	NN	O	B-protein
virus	NN	O	I-protein
early	NN	O	I-protein
antigen	NN	O	I-protein
diffuse	NN	O	I-protein
component	NN	O	I-protein
(	NN	O	O
EA-D	NN	O	B-protein
)	NN	O	O
is	NN	O	O
essential	NN	O	O
for	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	B-DNA
DNA	NN	O	I-DNA
polymerase	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
its	NN	O	O
activity	NN	O	O
is	NN	O	O
suppressed	NN	O	O
during	NN	O	O
latent	NN	O	O
infection	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
the	NN	O	O
promoter	NN	O	B-DNA
(	NN	O	O
BMRF1	NN	O	B-DNA
)	NN	O	O
for	NN	O	O
this	NN	O	O
early	NN	O	O
gene	NN	O	O
by	NN	O	O
studying	NN	O	O
its	NN	O	O
responsiveness	NN	O	O
in	NN	O	O
vitro	NN	O	O
to	NN	O	O
two	NN	O	O
immediate-early	NN	O	B-protein
viral	NN	O	I-protein
transactivators	NN	O	I-protein
,	NN	O	O
BZLF1	NN	O	B-protein
(	NN	O	O
Z	NN	O	B-protein
)	NN	O	O
and	NN	O	O
BRLF1	NN	O	B-protein
(	NN	O	O
R	NN	O	B-protein
)	NN	O	O
,	NN	O	O
focusing	NN	O	O
on	NN	O	O
the	NN	O	O
differences	NN	O	O
in	NN	O	O
response	NN	O	O
in	NN	O	O
lymphoid	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

In	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
Z	NN	O	B-protein
or	NN	O	O
R	NN	O	B-protein
alone	NN	O	O
produced	NN	O	O
only	NN	O	O
small	NN	O	O
increases	NN	O	O
in	NN	O	O
EA-D	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
,	NN	O	O
whereas	NN	O	O
both	NN	O	O
transactivators	NN	O	B-protein
together	NN	O	O
produced	NN	O	O
a	NN	O	O
large	NN	O	O
stimulatory	NN	O	O
effect	NN	O	O
.	NN	O	O

In	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
Z	NN	O	B-protein
transactivator	NN	O	I-protein
alone	NN	O	O
produced	NN	O	O
maximal	NN	O	O
stimulation	NN	O	O
of	NN	O	O
the	NN	O	O
EA-D	NN	O	B-DNA
promoter	NN	O	I-DNA
;	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
R	NN	O	B-protein
and	NN	O	O
Z	NN	O	B-protein
together	NN	O	O
was	NN	O	O
no	NN	O	O
greater	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
Z	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

Deletional	NN	O	O
analysis	NN	O	O
and	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
EA-D	NN	O	B-DNA
promoter	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
in	NN	O	O
epithelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
the	NN	O	O
potential	NN	O	O
AP-1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
plays	NN	O	O
an	NN	O	O
essential	NN	O	O
role	NN	O	O
in	NN	O	O
Z	NN	O	B-protein
responsiveness	NN	O	O
,	NN	O	O
although	NN	O	O
sequences	NN	O	O
further	NN	O	O
upstream	NN	O	O
are	NN	O	O
also	NN	O	O
important	NN	O	O
.	NN	O	O

In	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
only	NN	O	O
the	NN	O	O
upstream	NN	O	B-DNA
sequences	NN	O	I-DNA
are	NN	O	O
required	NN	O	O
for	NN	O	O
transactivation	NN	O	O
by	NN	O	O
the	NN	O	O
Z/R	NN	O	B-protein
combination	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
site	NN	O	I-protein
is	NN	O	O
dispensable	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
EA-D	NN	O	B-DNA
(	NN	O	I-DNA
BMRF1	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
regulation	NN	O	O
by	NN	O	O
Z	NN	O	B-protein
and	NN	O	O
R	NN	O	B-protein
is	NN	O	O
cell	NN	O	O
type	NN	O	O
specific	NN	O	O
and	NN	O	O
appears	NN	O	O
to	NN	O	O
involve	NN	O	O
different	NN	O	O
mechanisms	NN	O	O
in	NN	O	O
each	NN	O	O
cell	NN	O	O
type	NN	O	O
.	NN	O	O

-DOCSTART-	O

Complementary	NN	O	B-DNA
DNA	NN	O	I-DNA
encoding	NN	O	O
the	NN	O	O
human	NN	O	B-protein
T-cell	NN	O	I-protein
FK506-binding	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
a	NN	O	O
peptidylprolyl	NN	O	B-protein
cis-trans	NN	O	I-protein
isomerase	NN	O	I-protein
distinct	NN	O	O
from	NN	O	O
cyclophilin	NN	O	B-protein
.	NN	O	O

The	NN	O	O
recently	NN	O	O
discovered	NN	O	O
macrolide	NN	O	O
FK506	NN	O	O
has	NN	O	O
been	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
have	NN	O	O
potent	NN	O	O
immunosuppressive	NN	O	O
activity	NN	O	O
at	NN	O	O
concentrations	NN	O	O
100-fold	NN	O	O
lower	NN	O	O
than	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
a	NN	O	O
cyclic	NN	O	O
undecapeptide	NN	O	O
that	NN	O	O
is	NN	O	O
used	NN	O	O
to	NN	O	O
prevent	NN	O	O
rejection	NN	O	O
after	NN	O	O
transplantation	NN	O	O
of	NN	O	O
bone	NN	O	O
marrow	NN	O	O
and	NN	O	O
organs	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
kidney	NN	O	O
,	NN	O	O
heart	NN	O	O
,	NN	O	O
and	NN	O	O
liver	NN	O	O
.	NN	O	O

After	NN	O	O
the	NN	O	O
recent	NN	O	O
discovery	NN	O	O
that	NN	O	O
the	NN	O	O
cyclosporin	NN	O	B-protein
A-binding	NN	O	I-protein
protein	NN	O	I-protein
cyclophilin	NN	O	B-protein
is	NN	O	O
identical	NN	O	O
to	NN	O	O
peptidylprolyl	NN	O	B-protein
cis-trans	NN	O	I-protein
isomerase	NN	O	I-protein
,	NN	O	O
a	NN	O	O
cellular	NN	O	B-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
for	NN	O	O
FK506	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
distinct	NN	O	O
from	NN	O	O
cyclophilin	NN	O	B-protein
but	NN	O	O
to	NN	O	O
have	NN	O	O
the	NN	O	O
same	NN	O	O
enzymatic	NN	O	O
activity	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
isolated	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
coding	NN	O	O
for	NN	O	O
FK506-binding	NN	O	O
protein	NN	O	O
(	NN	O	O
FKBP	NN	O	B-protein
)	NN	O	O
from	NN	O	O
human	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
using	NN	O	O
mixed	NN	O	O
20-mer	NN	O	O
oligonucleotide	NN	O	O
probes	NN	O	O
synthesized	NN	O	O
on	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
sequence	NN	O	O
,	NN	O	O
Glu-Asp-Gly-Lys-Lys-Phe-Asp	NN	O	O
,	NN	O	O
reported	NN	O	O
for	NN	O	O
bovine	NN	O	B-protein
FKBP	NN	O	I-protein
.	NN	O	O

The	NN	O	O
DNA	NN	O	O
isolated	NN	O	O
contained	NN	O	O
an	NN	O	O
open	NN	O	O
reading	NN	O	O
frame	NN	O	O
encoding	NN	O	O
108	NN	O	O
amino	NN	O	O
acid	NN	O	O
residues	NN	O	O
.	NN	O	O

The	NN	O	O
first	NN	O	O
40	NN	O	O
residues	NN	O	O
of	NN	O	O
the	NN	O	O
deduced	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
were	NN	O	O
identical	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
the	NN	O	O
reported	NN	O	O
amino-terminal	NN	O	O
sequence	NN	O	O
of	NN	O	O
bovine	NN	O	O
FKBP	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
isolated	NN	O	O
represents	NN	O	O
the	NN	O	O
gene	NN	O	O
coding	NN	O	O
for	NN	O	O
FKBP	NN	O	B-protein
.	NN	O	O

Computer-assisted	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
deduced	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
indicates	NN	O	O
that	NN	O	O
FKBP	NN	O	B-protein
exhibits	NN	O	O
no	NN	O	O
internal	NN	O	O
homology	NN	O	O
and	NN	O	O
does	NN	O	O
not	NN	O	O
have	NN	O	O
significant	NN	O	O
sequence	NN	O	O
similarity	NN	O	O
to	NN	O	O
any	NN	O	O
other	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequences	NN	O	O
of	NN	O	O
known	NN	O	O
proteins	NN	O	O
,	NN	O	O
including	NN	O	O
cyclophilin	NN	O	B-protein
.	NN	O	O

This	NN	O	O
result	NN	O	O
suggests	NN	O	O
that	NN	O	O
two	NN	O	O
catalytically	NN	O	O
similar	NN	O	O
proteins	NN	O	O
,	NN	O	O
cyclophilin	NN	O	B-protein
and	NN	O	O
FKBP	NN	O	B-protein
,	NN	O	O
evolved	NN	O	O
independently	NN	O	O
.	NN	O	O

In	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
mRNA	NN	O	B-RNA
species	NN	O	I-RNA
of	NN	O	O
approximately	NN	O	O
1.8	NN	O	O
kilobases	NN	O	O
that	NN	O	O
hybridized	NN	O	O
with	NN	O	O
human	NN	O	B-DNA
FKBP	NN	O	I-DNA
cDNA	NN	O	I-DNA
were	NN	O	O
detected	NN	O	O
in	NN	O	O
poly	NN	O	B-RNA
(	NN	O	I-RNA
A	NN	O	I-RNA
)	NN	O	I-RNA
+	NN	O	I-RNA
RNAs	NN	O	I-RNA
from	NN	O	O
brain	NN	O	O
,	NN	O	O
lung	NN	O	O
,	NN	O	O
liver	NN	O	O
,	NN	O	O
and	NN	O	O
placental	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
leukocytes	NN	O	B-cell_type
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
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
did	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
FKBP	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Southern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
human	NN	O	B-DNA
genomic	NN	O	I-DNA
DNA	NN	O	I-DNA
digested	NN	O	O
with	NN	O	O
different	NN	O	O
restriction	NN	O	B-protein
enzymes	NN	O	I-protein
suggests	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
only	NN	O	O
a	NN	O	O
few	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
sequence	NN	O	O
encoding	NN	O	O
FKBP	NN	O	B-protein
.	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
result	NN	O	O
that	NN	O	O
as	NN	O	O
many	NN	O	O
as	NN	O	O
20	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
cyclophilin	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
possible	NN	O	O
pseudogenes	NN	O	O
may	NN	O	O
be	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
mammalian	NN	O	B-DNA
genome	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Involvement	NN	O	O
of	NN	O	O
a	NN	O	O
second	NN	O	O
lymphoid-specific	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy-chain	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
whether	NN	O	O
enhancer	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
highly	NN	O	O
conserved	NN	O	O
octamer	NN	O	B-DNA
(	NN	O	I-DNA
OCTA	NN	O	I-DNA
)	NN	O	I-DNA
-nucleotide	NN	O	I-DNA
motif	NN	O	I-DNA
are	NN	O	O
important	NN	O	O
for	NN	O	O
lymphoid-specific	NN	O	O
expression	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
gene	NN	O	I-DNA
,	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
mutating	NN	O	O
the	NN	O	O
binding	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
additional	NN	O	I-protein
lymphoid-specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
designated	NN	O	O
NF-microB	NN	O	B-protein
,	NN	O	O
in	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
IgH	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
NF-microB-binding	NN	O	B-DNA
site	NN	O	I-DNA
plays	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
in	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
because	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
microB	NN	O	O
DNA	NN	O	O
motif	NN	O	O
decreased	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell	NN	O	B-cell_type
lineage	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
nonlymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

This	NN	O	O
effect	NN	O	O
was	NN	O	O
comparable	NN	O	O
to	NN	O	O
or	NN	O	O
even	NN	O	O
stronger	NN	O	O
than	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
a	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
OCTA	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
combined	NN	O	O
mutation	NN	O	O
of	NN	O	O
both	NN	O	O
microB	NN	O	B-DNA
and	NN	O	O
OCTA	NN	O	B-DNA
sites	NN	O	I-DNA
further	NN	O	O
reduced	NN	O	O
enhancer	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

Interestingly	NN	O	O
,	NN	O	O
alteration	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
microB	NN	O	B-DNA
or	NN	O	O
E3	NN	O	B-DNA
site	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
70-base-pair	NN	O	B-DNA
fragment	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
that	NN	O	O
lacks	NN	O	O
the	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
OCTA	NN	O	O
abolished	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
in	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
completely	NN	O	O
.	NN	O	O

Nevertheless	NN	O	O
,	NN	O	O
a	NN	O	O
multimer	NN	O	O
of	NN	O	O
the	NN	O	O
microB	NN	O	B-DNA
motif	NN	O	I-DNA
alone	NN	O	O
showed	NN	O	O
no	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
.	NN	O	O

DNase	NN	O	O
footprinting	NN	O	O
analysis	NN	O	O
corroborated	NN	O	O
the	NN	O	O
functional	NN	O	O
data	NN	O	O
showing	NN	O	O
that	NN	O	O
a	NN	O	O
lymphoid-specific	NN	O	B-protein
protein	NN	O	I-protein
binds	NN	O	O
to	NN	O	O
the	NN	O	O
microB	NN	O	B-DNA
DNA	NN	O	I-DNA
motif	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
microB	NN	O	B-DNA
element	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
new	NN	O	O
crucial	NN	O	O
element	NN	O	O
important	NN	O	O
for	NN	O	O
lymphoid-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IgH	NN	O	O
gene	NN	O	O
but	NN	O	O
that	NN	O	O
interaction	NN	O	O
with	NN	O	O
another	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
is	NN	O	O
essential	NN	O	O
for	NN	O	O
its	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Stimulation	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
2	NN	O	O
(	NN	O	O
HIV-2	NN	O	O
)	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
the	NN	O	O
cytomegalovirus	NN	O	O
and	NN	O	O
HIV-2	NN	O	B-DNA
transactivator	NN	O	I-DNA
gene	NN	O	I-DNA
.	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
often	NN	O	O
causes	NN	O	O
latent	NN	O	O
infection	NN	O	O
.	NN	O	O

Transactivation	NN	O	O
by	NN	O	O
some	NN	O	O
DNA	NN	O	O
viruses	NN	O	O
has	NN	O	O
been	NN	O	O
implicated	NN	O	O
in	NN	O	O
inducing	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
and	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

The	NN	O	O
transactivator	NN	O	B-DNA
(	NN	O	I-DNA
IE-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
of	NN	O	O
the	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
enhance	NN	O	O
HIV-2	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

This	NN	O	O
inducer	NN	O	O
can	NN	O	O
act	NN	O	O
in	NN	O	O
concert	NN	O	O
with	NN	O	O
the	NN	O	O
HIV-2	NN	O	B-DNA
tat	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
in	NN	O	O
enhancing	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

While	NN	O	O
the	NN	O	O
HIV-2	NN	O	O
and	NN	O	O
HIV-1	NN	O	B-DNA
tat	NN	O	I-DNA
genes	NN	O	I-DNA
and	NN	O	O
T-cell	NN	O	B-protein
activators	NN	O	I-protein
apparently	NN	O	O
employ	NN	O	O
independent	NN	O	O
modes	NN	O	O
of	NN	O	O
action	NN	O	O
,	NN	O	O
the	NN	O	O
CMV	NN	O	B-DNA
transactivator	NN	O	I-DNA
in	NN	O	O
combination	NN	O	O
with	NN	O	O
the	NN	O	O
HIV-2	NN	O	B-DNA
tat	NN	O	I-DNA
or	NN	O	O
T-cell	NN	O	B-protein
activators	NN	O	I-protein
may	NN	O	O
employ	NN	O	O
a	NN	O	O
gene	NN	O	O
activation	NN	O	O
pathway	NN	O	O
with	NN	O	O
some	NN	O	O
common	NN	O	O
and	NN	O	O
some	NN	O	O
distinct	NN	O	O
components	NN	O	O
.	NN	O	O

Both	NN	O	O
HIV-2	NN	O	O
and	NN	O	O
CMV	NN	O	B-DNA
transactivators	NN	O	I-DNA
enhance	NN	O	O
HIV-2	NN	O	O
gene	NN	O	O
expression	NN	O	O
by	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
involving	NN	O	O
transcript	NN	O	O
initiation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
elongation	NN	O	O
,	NN	O	O
with	NN	O	O
CMV	NN	O	B-DNA
transactivator	NN	O	I-DNA
affecting	NN	O	O
elongation	NN	O	O
more	NN	O	O
than	NN	O	O
the	NN	O	O
initiation	NN	O	O
.	NN	O	O

A	NN	O	O
significant	NN	O	O
proportion	NN	O	O
of	NN	O	O
transcripts	NN	O	B-RNA
appear	NN	O	O
to	NN	O	O
terminate	NN	O	O
prematurely	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
transactivators	NN	O	B-protein
.	NN	O	O

Deletion	NN	O	O
mutation	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-2	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
suggests	NN	O	O
that	NN	O	O
the	NN	O	O
element	NN	O	O
that	NN	O	O
responds	NN	O	O
to	NN	O	O
CMV	NN	O	O
transactivation	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
CD4+	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
either	NN	O	O
a	NN	O	O
diffuse	NN	O	O
one	NN	O	O
or	NN	O	O
located	NN	O	O
downstream	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-2	NN	O	B-DNA
enhancer	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

-DOCSTART-	O

Quantitative	NN	O	O
immunohistochemical	NN	O	O
analysis	NN	O	O
of	NN	O	O
mononuclear	NN	O	B-cell_type
infiltrates	NN	O	I-cell_type
in	NN	O	O
breast	NN	O	O
carcinomas	NN	O	O
--	NN	O	O
correlation	NN	O	O
with	NN	O	O
tumour	NN	O	O
differentiation	NN	O	O
.	NN	O	O

Inflammatory	NN	O	B-cell_type
infiltrates	NN	O	I-cell_type
were	NN	O	O
analysed	NN	O	O
in	NN	O	O
tissue	NN	O	O
sections	NN	O	O
of	NN	O	O
76	NN	O	O
breast	NN	O	O
carcinomas	NN	O	O
by	NN	O	O
counting	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
macrophages	NN	O	B-cell_type
,	NN	O	O
IgA+	NN	O	B-cell_type
and	NN	O	I-cell_type
IgG+	NN	O	I-cell_type
plasma	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
their	NN	O	O
subpopulations	NN	O	O
,	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
and	NN	O	O
by	NN	O	O
measuring	NN	O	O
postcapillary	NN	O	O
venules	NN	O	O
(	NN	O	O
PCVs	NN	O	O
,	NN	O	O
found	NN	O	O
in	NN	O	O
12	NN	O	O
cases	NN	O	O
)	NN	O	O
within	NN	O	O
the	NN	O	O
infiltrates	NN	O	O
.	NN	O	O

These	NN	O	O
parameters	NN	O	O
were	NN	O	O
correlated	NN	O	O
with	NN	O	O
nuclear	NN	O	O
grade	NN	O	O
and	NN	O	O
biochemically	NN	O	O
determined	NN	O	O
hormone	NN	O	O
receptor	NN	O	O
status	NN	O	O
,	NN	O	O
known	NN	O	O
markers	NN	O	O
of	NN	O	O
tumour	NN	O	O
differentiation	NN	O	O
.	NN	O	O

A	NN	O	O
direct	NN	O	O
correlation	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
inflammation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
grade	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.0001	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
an	NN	O	O
inverse	NN	O	O
correlation	NN	O	O
between	NN	O	O
inflammation	NN	O	O
and	NN	O	O
oestrogen	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
OR	NN	O	B-protein
)	NN	O	O
positivity	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
as	NN	O	O
well	NN	O	O
as	NN	O	O
inflammation	NN	O	O
and	NN	O	O
progesterone	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
PR	NN	O	B-protein
)	NN	O	O
positivity	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
percentage	NN	O	O
of	NN	O	O
the	NN	O	O
OKT8+	NN	O	B-cell_line
suppressor/cytotoxic	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
increased	NN	O	O
when	NN	O	O
the	NN	O	O
inflammation	NN	O	O
expanded	NN	O	O
from	NN	O	O
scanty	NN	O	O
to	NN	O	O
moderate	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.02	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
diameter	NN	O	O
of	NN	O	O
the	NN	O	O
PCVs	NN	O	O
also	NN	O	O
increased	NN	O	O
with	NN	O	O
increasing	NN	O	O
inflammatory	NN	O	O
infiltrate	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.02	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
a	NN	O	O
direct	NN	O	O
correlation	NN	O	O
exists	NN	O	O
between	NN	O	O
the	NN	O	O
diameter	NN	O	O
of	NN	O	O
the	NN	O	O
PCVs	NN	O	O
and	NN	O	O
both	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
the	NN	O	O
OKT8+	NN	O	B-cell_line
T	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
0.04	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
Leu-7+	NN	O	O
natural	NN	O	B-cell_type
killer	NN	O	I-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.03	NN	O	O
)	NN	O	O
.	NN	O	O

-DOCSTART-	O

Reactivity	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_line
to	NN	O	O
a	NN	O	O
progesterone	NN	O	B-protein
receptor-specific	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
.	NN	O	O

In	NN	O	O
this	NN	O	O
study	NN	O	O
we	NN	O	O
present	NN	O	O
evidence	NN	O	O
for	NN	O	O
reactivity	NN	O	O
of	NN	O	O
pregnancy	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
not	NN	O	O
nonpregnancy	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
with	NN	O	O
the	NN	O	O
progesterone	NN	O	B-protein
receptor-specific	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
mPRI	NN	O	B-protein
.	NN	O	O

Using	NN	O	O
an	NN	O	O
avidin	NN	O	B-protein
-biotin	NN	O	B-protein
peroxidase	NN	O	I-protein
detection	NN	O	O
system	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
a	NN	O	O
nuclear	NN	O	O
staining	NN	O	O
in	NN	O	O
14.6	NN	O	O
+/-	NN	O	O
3.7	NN	O	O
%	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
27	NN	O	O
)	NN	O	O
of	NN	O	O
pregnancy	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
,	NN	O	O
while	NN	O	O
only	NN	O	O
0.47	NN	O	O
+/-	NN	O	O
0.33	NN	O	O
%	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
15	NN	O	O
)	NN	O	O
of	NN	O	O
nonpregnancy	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
reacted	NN	O	O
with	NN	O	O
the	NN	O	O
antibody	NN	O	O
.	NN	O	O

To	NN	O	O
characterize	NN	O	O
the	NN	O	O
receptor-bearing	NN	O	O
subset	NN	O	O
,	NN	O	O
CD8+	NN	O	B-cell_line
and	NN	O	I-cell_line
CD4+	NN	O	I-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
depleted	NN	O	O
by	NN	O	O
complement-dependent	NN	O	O
lysis	NN	O	O
.	NN	O	O

Depletion	NN	O	O
of	NN	O	O
CD8+	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
62	NN	O	O
+/-	NN	O	O
18	NN	O	O
%	NN	O	O
loss	NN	O	O
of	NN	O	O
progesterone	NN	O	B-cell_line
receptor-bearing	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
depletion	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_line
cells	NN	O	I-cell_line
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
number	NN	O	O
of	NN	O	O
positively	NN	O	O
staining	NN	O	O
lymphocytes	NN	O	B-cell_line
.	NN	O	O

In	NN	O	O
nonpregnancy	NN	O	B-cell_line
lymphocytes	NN	O	I-cell_line
a	NN	O	O
3-day	NN	O	O
PHA	NN	O	B-protein
treatment	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
allogeneic	NN	O	O
stimulation	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	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
receptor-containing	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
pregnancy	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
nonpregnancy	NN	O	O
,	NN	O	O
lymphocytes	NN	O	B-cell_line
contain	NN	O	O
progesterone	NN	O	B-protein
binding	NN	O	I-protein
structures	NN	O	I-protein
,	NN	O	O
and	NN	O	O
that	NN	O	O
these	NN	O	O
are	NN	O	O
inducible	NN	O	O
by	NN	O	O
mitogenic	NN	O	O
or	NN	O	O
alloantigenic	NN	O	O
stimuli	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	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
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Interaction	NN	O	O
of	NN	O	O
fibronectin	NN	O	B-protein
with	NN	O	O
VLA-5	NN	O	B-protein
receptor	NN	O	I-protein
on	NN	O	O
CD4	NN	O	B-cell_line
cells	NN	O	I-cell_line
induces	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

Fibronectin	NN	O	B-protein
synergized	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
antibody	NN	O	I-protein
to	NN	O	O
promote	NN	O	O
CD4	NN	O	B-protein
cell	NN	O	O
proliferation	NN	O	O
in	NN	O	O
a	NN	O	O
serum-free	NN	O	O
culture	NN	O	O
system	NN	O	O
whereas	NN	O	O
no	NN	O	O
proliferation	NN	O	O
was	NN	O	O
observed	NN	O	O
when	NN	O	O
CD4	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
cultured	NN	O	O
with	NN	O	O
anti-CD3	NN	O	B-protein
alone	NN	O	O
or	NN	O	O
fibronectin	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
anti-CD29	NN	O	B-protein
(	NN	O	O
integrin	NN	O	B-protein
beta	NN	O	I-protein
1	NN	O	I-protein
)	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
anti-VLA-5	NN	O	B-protein
(	NN	O	O
human	NN	O	B-protein
fibronectin	NN	O	I-protein
receptor	NN	O	I-protein
)	NN	O	O
antibodies	NN	O	O
blocked	NN	O	O
this	NN	O	O
CD4	NN	O	B-cell_line
cell	NN	O	I-cell_line
activation	NN	O	O
in	NN	O	O
this	NN	O	O
system	NN	O	O
.	NN	O	O

Although	NN	O	O
anti-CD3	NN	O	B-protein
alone	NN	O	O
or	NN	O	O
fibronectin	NN	O	B-protein
alone	NN	O	O
can	NN	O	O
not	NN	O	O
induce	NN	O	O
IL-2	NN	O	B-protein
message	NN	O	O
by	NN	O	O
CD4	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
combination	NN	O	O
of	NN	O	O
anti-CD3	NN	O	B-protein
plus	NN	O	O
fibronectin	NN	O	B-protein
induced	NN	O	O
IL-2	NN	O	B-protein
message	NN	O	O
by	NN	O	O
CD4	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
an	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
molecular	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
IL-2	NN	O	B-protein
message	NN	O	O
was	NN	O	O
generated	NN	O	O
,	NN	O	O
we	NN	O	O
showed	NN	O	O
that	NN	O	O
a	NN	O	O
fibronectin	NN	O	B-protein
-VLA-5	NN	O	O
fibronectin	NN	O	B-protein
receptor	NN	O	I-protein
interaction	NN	O	O
may	NN	O	O
contribute	NN	O	O
an	NN	O	O
independent	NN	O	O
signal	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
CD3	NN	O	B-protein
pathway	NN	O	O
of	NN	O	O
activation	NN	O	O
by	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
an	NN	O	O
AP-1	NN	O	B-protein
transcriptional	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
the	NN	O	O
VLA-5	NN	O	B-protein
fibronectin	NN	O	I-protein
receptor	NN	O	I-protein
on	NN	O	O
CD4	NN	O	B-cell_line
cells	NN	O	I-cell_line
can	NN	O	O
play	NN	O	O
a	NN	O	O
complementary	NN	O	O
role	NN	O	O
in	NN	O	O
CD3	NN	O	B-protein
-	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
its	NN	O	O
interaction	NN	O	O
with	NN	O	O
fibronectin	NN	O	B-protein
.	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
Alzheimer	NN	O	O
's	NN	O	O
disease	NN	O	O
.	NN	O	O

Several	NN	O	O
lines	NN	O	O
of	NN	O	O
evidence	NN	O	O
suggest	NN	O	O
disturbances	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
system	NN	O	O
in	NN	O	O
Alzheimer	NN	O	O
's	NN	O	O
disease	NN	O	O
(	NN	O	O
AD	NN	O	O
)	NN	O	O
.	NN	O	O

In	NN	O	O
an	NN	O	O
exploration	NN	O	O
of	NN	O	O
the	NN	O	O
potential	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	O
receptor	NN	O	O
(	NN	O	O
GR	NN	O	O
)	NN	O	O
in	NN	O	O
AD	NN	O	O
,	NN	O	O
GR	NN	O	O
density	NN	O	O
and	NN	O	O
affinity	NN	O	O
were	NN	O	O
assessed	NN	O	O
on	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
of	NN	O	O
12	NN	O	O
AD	NN	O	O
patients	NN	O	O
and	NN	O	O
12	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

GR	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
between	NN	O	O
patients	NN	O	O
and	NN	O	O
controls	NN	O	O
or	NN	O	O
between	NN	O	O
patients	NN	O	O
subdivided	NN	O	O
according	NN	O	O
to	NN	O	O
diagnosis	NN	O	O
or	NN	O	O
associated	NN	O	O
clinical	NN	O	O
features	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
abnormalities	NN	O	O
of	NN	O	O
the	NN	O	O
HPA	NN	O	O
system	NN	O	O
in	NN	O	O
AD	NN	O	O
are	NN	O	O
not	NN	O	O
related	NN	O	O
to	NN	O	O
a	NN	O	O
GR	NN	O	B-protein
deficiency	NN	O	O
.	NN	O	O

-DOCSTART-	O

Oncogene	NN	O	O
amplification	NN	O	O
correlates	NN	O	O
with	NN	O	O
dense	NN	O	O
lymphocyte	NN	O	O
infiltration	NN	O	O
in	NN	O	O
human	NN	O	O
breast	NN	O	O
cancers	NN	O	O
:	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
hematopoietic	NN	O	B-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
release	NN	O	O
by	NN	O	O
tumor	NN	O	B-cell_type
cells	NN	O	I-cell_type
?	NN	O	O

One	NN	O	O
hundred	NN	O	O
six	NN	O	O
primary	NN	O	O
breast	NN	O	O
cancer	NN	O	O
samples	NN	O	O
were	NN	O	O
analysed	NN	O	O
for	NN	O	O
c-erbB2	NN	O	O
,	NN	O	O
int-2	NN	O	O
,	NN	O	O
and	NN	O	O
c-myc	NN	O	O
gene	NN	O	O
amplification	NN	O	O
.	NN	O	O

Surgically	NN	O	O
confirmed	NN	O	O
nodal	NN	O	O
involvement	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
42	NN	O	O
%	NN	O	O
.	NN	O	O

Level	NN	O	O
of	NN	O	O
gene	NN	O	O
amplification	NN	O	O
was	NN	O	O
studied	NN	O	O
by	NN	O	O
Southern	NN	O	O
and/or	NN	O	O
slot	NN	O	O
blot	NN	O	O
techniques	NN	O	O
.	NN	O	O

Amplified	NN	O	O
c-erbB2	NN	O	B-DNA
gene	NN	O	I-DNA
sequences	NN	O	I-DNA
were	NN	O	O
present	NN	O	O
in	NN	O	O
21.5	NN	O	O
%	NN	O	O
of	NN	O	O
all	NN	O	O
samples	NN	O	O
.	NN	O	O

Int-2	NN	O	B-DNA
was	NN	O	O
amplified	NN	O	O
in	NN	O	O
13.1	NN	O	O
%	NN	O	O
and	NN	O	O
c-myc	NN	O	B-DNA
was	NN	O	O
amplified	NN	O	O
in	NN	O	O
10.3	NN	O	O
%	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
non-parametric	NN	O	O
test	NN	O	O
(	NN	O	O
Kruskal-Wallis	NN	O	O
)	NN	O	O
a	NN	O	O
strong	NN	O	O
negative	NN	O	O
association	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
c-erbB2	NN	O	B-DNA
amplification	NN	O	O
and	NN	O	O
absence	NN	O	O
of	NN	O	O
estrogen	NN	O	O
receptor	NN	O	O
(	NN	O	O
ER	NN	O	O
)	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.0009	NN	O	O
)	NN	O	O
or	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
P	NN	O	O
=	NN	O	O
.011	NN	O	O
)	NN	O	O
expression	NN	O	O
.	NN	O	O

No	NN	O	O
correlations	NN	O	O
were	NN	O	O
found	NN	O	O
between	NN	O	O
all	NN	O	O
or	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
amplification	NN	O	O
of	NN	O	O
each	NN	O	O
oncogene	NN	O	O
separately	NN	O	O
or	NN	O	O
combined	NN	O	O
with	NN	O	O
T	NN	O	O
,	NN	O	O
N	NN	O	O
,	NN	O	O
grade	NN	O	O
,	NN	O	O
multifocality	NN	O	O
of	NN	O	O
tumor	NN	O	O
,	NN	O	O
or	NN	O	O
associated	NN	O	O
carcinoma	NN	O	O
in	NN	O	O
situ	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
trend	NN	O	O
approaching	NN	O	O
statistical	NN	O	O
significance	NN	O	O
for	NN	O	O
patients	NN	O	O
with	NN	O	O
c-erbB2	NN	O	B-DNA
amplifications	NN	O	O
to	NN	O	O
have	NN	O	O
positive	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
at	NN	O	O
surgery	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.09	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
somewhat	NN	O	O
surprising	NN	O	O
finding	NN	O	O
however	NN	O	O
was	NN	O	O
a	NN	O	O
very	NN	O	O
strong	NN	O	O
association	NN	O	O
between	NN	O	O
oncogene	NN	O	B-DNA
amplification	NN	O	O
and	NN	O	O
dense	NN	O	O
lymphocyte	NN	O	O
infiltration	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
.05	NN	O	O
)	NN	O	O
.	NN	O	O
This	NN	O	O
correlation	NN	O	O
is	NN	O	O
even	NN	O	O
stronger	NN	O	O
when	NN	O	O
only	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
amplification	NN	O	O
are	NN	O	O
considered	NN	O	O
,	NN	O	O
either	NN	O	O
for	NN	O	O
each	NN	O	O
oncogene	NN	O	O
separately	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.0048	NN	O	O
)	NN	O	O
or	NN	O	O
in	NN	O	O
combination	NN	O	O
(	NN	O	O
P	NN	O	O
=	NN	O	O
.0007	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
malignant	NN	O	O
cell	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
may	NN	O	O
help	NN	O	O
explain	NN	O	O
this	NN	O	O
observation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Suppression	NN	O	O
of	NN	O	O
signals	NN	O	O
required	NN	O	O
for	NN	O	O
activation	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
in	NN	O	O
cells	NN	O	O
constitutively	NN	O	O
expressing	NN	O	O
the	NN	O	O
HTLV-I	NN	O	B-protein
Tax	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Transient	NN	O	O
short-term	NN	O	O
expression	NN	O	O
of	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-I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
leads	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
pleiotropic	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

Consistent	NN	O	O
with	NN	O	O
findings	NN	O	O
obtained	NN	O	O
with	NN	O	O
transient	NN	O	O
expression	NN	O	O
assays	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
marked	NN	O	O
accumulation	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
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
of	NN	O	O
Namalwa	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
constitutively	NN	O	O
express	NN	O	O
Tax	NN	O	B-protein
.	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
activity	NN	O	O
was	NN	O	O
not	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
following	NN	O	O
long-term	NN	O	O
expression	NN	O	O
of	NN	O	O
Tax	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
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
both	NN	O	O
mitogens	NN	O	O
and	NN	O	O
cytokines	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
was	NN	O	O
also	NN	O	O
blocked	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
constitutively	NN	O	O
expressing	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
other	NN	O	O
mitogen-inducible	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
,	NN	O	O
was	NN	O	O
unaffected	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
depending	NN	O	O
on	NN	O	O
the	NN	O	O
cellular	NN	O	O
environment	NN	O	O
,	NN	O	O
the	NN	O	O
short-	NN	O	O
and	NN	O	O
long-term	NN	O	O
effects	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
expression	NN	O	O
can	NN	O	O
be	NN	O	O
quite	NN	O	O
different	NN	O	O
.	NN	O	O

Consequently	NN	O	O
,	NN	O	O
one	NN	O	O
function	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
in	NN	O	O
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
HTLV-I	NN	O	O
might	NN	O	O
involve	NN	O	O
cell-type-specific	NN	O	O
suppression	NN	O	O
,	NN	O	O
as	NN	O	O
opposed	NN	O	O
to	NN	O	O
activation	NN	O	O
,	NN	O	O
of	NN	O	O
distinct	NN	O	O
signal	NN	O	O
pathways	NN	O	O
.	NN	O	O

The	NN	O	O
cells	NN	O	O
lines	NN	O	O
described	NN	O	O
here	NN	O	O
should	NN	O	O
be	NN	O	O
useful	NN	O	O
for	NN	O	O
the	NN	O	O
delineation	NN	O	O
of	NN	O	O
signaling	NN	O	O
pathways	NN	O	O
utilized	NN	O	O
in	NN	O	O
the	NN	O	O
selective	NN	O	O
regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interferon-gamma	NN	O	B-protein
and	NN	O	O
the	NN	O	O
sexual	NN	O	O
dimorphism	NN	O	O
of	NN	O	O
autoimmunity	NN	O	O
.	NN	O	O

The	NN	O	O
sexual	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
incidence	NN	O	O
of	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
has	NN	O	O
remained	NN	O	O
an	NN	O	O
enigma	NN	O	O
for	NN	O	O
many	NN	O	O
years	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
examination	NN	O	O
of	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
autoimmunity	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
,	NN	O	O
evidence	NN	O	O
has	NN	O	O
been	NN	O	O
obtained	NN	O	O
further	NN	O	O
implicating	NN	O	O
the	NN	O	O
lymphokine	NN	O	B-protein
interferon-gamma	NN	O	B-protein
in	NN	O	O
the	NN	O	O
etiology	NN	O	O
of	NN	O	O
autoimmunity	NN	O	O
.	NN	O	O

Sex	NN	O	O
steroid	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
this	NN	O	O
molecule	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
other	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
may	NN	O	O
help	NN	O	O
explain	NN	O	O
the	NN	O	O
gender-specific	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
,	NN	O	O
including	NN	O	O
autoimmunity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
of	NN	O	O
a	NN	O	O
transcriptionally	NN	O	B-protein
active	NN	O	I-protein
human	NN	O	I-protein
TATA	NN	O	I-protein
binding	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Transcription	NN	O	B-protein
factor	NN	O	I-protein
IID	NN	O	I-protein
(	NN	O	O
TFIID	NN	O	B-protein
)	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
TATA	NN	O	B-DNA
box	NN	O	I-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
and	NN	O	O
regulates	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
most	NN	O	O
eukaryotic	NN	O	B-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

Complementary	NN	O	B-DNA
DNA	NN	O	I-DNA
(	NN	O	O
cDNA	NN	O	B-DNA
)	NN	O	O
encoding	NN	O	O
a	NN	O	O
human	NN	O	B-protein
TFIID	NN	O	I-protein
protein	NN	O	I-protein
has	NN	O	O
been	NN	O	O
cloned	NN	O	O
.	NN	O	O

The	NN	O	O
human	NN	O	B-protein
TFIID	NN	O	I-protein
polypeptide	NN	O	I-protein
has	NN	O	O
339	NN	O	O
amino	NN	O	O
acids	NN	O	O
and	NN	O	O
a	NN	O	O
molecular	NN	O	O
size	NN	O	O
of	NN	O	O
37	NN	O	O
,	NN	O	O
745	NN	O	O
daltons	NN	O	O
.	NN	O	O

The	NN	O	O
carboxyl-terminal	NN	O	B-protein
181	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
TFIID	NN	O	I-protein
protein	NN	O	I-protein
shares	NN	O	O
80	NN	O	O
%	NN	O	O
identity	NN	O	O
with	NN	O	O
the	NN	O	O
TFIID	NN	O	B-protein
protein	NN	O	O
from	NN	O	O
Saccharomyces	NN	O	O
cerevisiae	NN	O	O
.	NN	O	O

The	NN	O	O
amino	NN	O	B-protein
terminus	NN	O	I-protein
contains	NN	O	O
an	NN	O	O
unusual	NN	O	O
repeat	NN	O	O
of	NN	O	O
38	NN	O	O
consecutive	NN	O	O
glutamine	NN	O	O
residues	NN	O	O
and	NN	O	O
an	NN	O	O
X-Thr-Pro	NN	O	B-protein
repeat	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
DNA	NN	O	O
in	NN	O	O
reticulocyte	NN	O	O
lysates	NN	O	O
or	NN	O	O
in	NN	O	O
Escherichia	NN	O	O
coli	NN	O	O
yielded	NN	O	O
a	NN	O	O
protein	NN	O	O
that	NN	O	O
was	NN	O	O
competent	NN	O	O
for	NN	O	O
both	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
transcription	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
T-cell	NN	O	B-protein
trans-activator	NN	O	I-protein
that	NN	O	O
recognizes	NN	O	O
a	NN	O	O
phorbol	NN	O	B-DNA
ester-inducible	NN	O	I-DNA
element	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
promoter	NN	O	I-DNA
.	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
promoter	NN	O	I-DNA
is	NN	O	O
recognized	NN	O	O
by	NN	O	O
several	NN	O	O
cell-type-specific	NN	O	B-protein
and	NN	O	I-protein
ubiquitous	NN	O	I-protein
transcriptional	NN	O	I-protein
regulators	NN	O	I-protein
that	NN	O	O
integrate	NN	O	O
information	NN	O	O
transmitted	NN	O	O
by	NN	O	O
various	NN	O	O
signaling	NN	O	O
systems	NN	O	O
leading	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
and	NN	O	O
T-cell	NN	O	B-cell_type
activation	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
transfection	NN	O	O
,	NN	O	O
protein-DNA	NN	O	O
binding	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
methods	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
discovered	NN	O	O
the	NN	O	O
novel	NN	O	B-protein
T-cell-specific	NN	O	I-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
TCF-1	NN	O	B-protein
(	NN	O	O
for	NN	O	O
T-Cell	NN	O	B-protein
Factor-1	NN	O	I-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
recognizes	NN	O	O
a	NN	O	O
T-cell-specific	NN	O	B-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
(	NN	O	O
TCE	NN	O	B-DNA
)	NN	O	O
located	NN	O	O
within	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
the	NN	O	O
TCE	NN	O	B-DNA
is	NN	O	O
similar	NN	O	O
in	NN	O	O
sequence	NN	O	O
to	NN	O	O
a	NN	O	O
consensus	NN	O	B-DNA
NF	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
site	NN	O	I-DNA
,	NN	O	O
several	NN	O	O
criteria	NN	O	O
indicate	NN	O	O
that	NN	O	O
TCF-1	NN	O	B-protein
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

However	NN	O	O
,	NN	O	O
like	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
TCF-1	NN	O	B-protein
activity	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
esters	NN	O	O
and	NN	O	O
other	NN	O	O
T-cell	NN	O	O
activators	NN	O	O
.	NN	O	O

-DOCSTART-	O

Risk	NN	O	O
factors	NN	O	O
for	NN	O	O
breast	NN	O	O
recurrence	NN	O	O
in	NN	O	O
premenopausal	NN	O	O
and	NN	O	O
postmenopausal	NN	O	O
patients	NN	O	O
with	NN	O	O
ductal	NN	O	O
cancers	NN	O	O
treated	NN	O	O
by	NN	O	O
conservation	NN	O	O
therapy	NN	O	O
.	NN	O	O

Risk	NN	O	O
factors	NN	O	O
for	NN	O	O
local	NN	O	O
failure	NN	O	O
were	NN	O	O
evaluated	NN	O	O
for	NN	O	O
496	NN	O	O
clinical	NN	O	O
Stage	NN	O	O
I-II	NN	O	O
patients	NN	O	O
with	NN	O	O
infiltrating	NN	O	O
ductal	NN	O	O
carcinomas	NN	O	O
(	NN	O	O
median	NN	O	O
follow-up	NN	O	O
,	NN	O	O
71	NN	O	O
months	NN	O	O
)	NN	O	O
treated	NN	O	O
by	NN	O	O
conservative	NN	O	O
surgery	NN	O	O
and	NN	O	O
radiotherapy	NN	O	O
.	NN	O	O

Monofactorial	NN	O	O
analysis	NN	O	O
identified	NN	O	O
the	NN	O	O
following	NN	O	O
factors	NN	O	O
to	NN	O	O
be	NN	O	O
correlated	NN	O	O
with	NN	O	O
increased	NN	O	O
risk	NN	O	O
:	NN	O	O
moderate/marked	NN	O	O
mononuclear	NN	O	O
cell	NN	O	O
reaction	NN	O	O
(	NN	O	O
MCR	NN	O	O
)	NN	O	O
,	NN	O	O
high	NN	O	O
histologic	NN	O	O
grade	NN	O	O
(	NN	O	O
G	NN	O	O
)	NN	O	O
,	NN	O	O
extensive	NN	O	O
intraductal	NN	O	O
component	NN	O	O
(	NN	O	O
EIC	NN	O	O
)	NN	O	O
,	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
,	NN	O	O
macroscopic	NN	O	O
multiplicity	NN	O	O
,	NN	O	O
estrogen	NN	O	B-protein
receptor	NN	O	I-protein
negativity	NN	O	O
,	NN	O	O
anatomic	NN	O	O
tumor	NN	O	O
size	NN	O	O
,	NN	O	O
age	NN	O	O
younger	NN	O	O
than	NN	O	O
40	NN	O	O
years	NN	O	O
,	NN	O	O
and	NN	O	O
vascular	NN	O	O
invasion	NN	O	O
.	NN	O	O

Only	NN	O	O
MCR	NN	O	O
,	NN	O	O
G	NN	O	O
,	NN	O	O
and	NN	O	O
EIC	NN	O	O
proved	NN	O	O
significant	NN	O	O
in	NN	O	O
Cox	NN	O	O
multivariate	NN	O	O
analysis	NN	O	O
.	NN	O	O

These	NN	O	O
risk	NN	O	O
factors	NN	O	O
were	NN	O	O
highly	NN	O	O
age	NN	O	O
dependent	NN	O	O
,	NN	O	O
with	NN	O	O
EIC	NN	O	O
markedly	NN	O	O
more	NN	O	O
prevalent	NN	O	O
in	NN	O	O
women	NN	O	O
younger	NN	O	O
than	NN	O	O
50	NN	O	O
,	NN	O	O
MCR	NN	O	O
and	NN	O	O
G	NN	O	O
in	NN	O	O
women	NN	O	O
younger	NN	O	O
than	NN	O	O
40	NN	O	O
.	NN	O	O

Separate	NN	O	O
Cox	NN	O	O
analysis	NN	O	O
for	NN	O	O
premenopausal	NN	O	O
patients	NN	O	O
showed	NN	O	O
that	NN	O	O
MCR/EIC	NN	O	O
determined	NN	O	O
risk	NN	O	O
independent	NN	O	O
of	NN	O	O
resection	NN	O	O
margins	NN	O	O
:	NN	O	O
tumors	NN	O	O
with	NN	O	O
MCR	NN	O	O
had	NN	O	O
a	NN	O	O
28	NN	O	O
%	NN	O	O
,	NN	O	O
and	NN	O	O
with	NN	O	O
EIC	NN	O	O
a	NN	O	O
22	NN	O	O
%	NN	O	O
probability	NN	O	O
of	NN	O	O
recurring	NN	O	O
locally	NN	O	O
by	NN	O	O
5	NN	O	O
years	NN	O	O
.	NN	O	O

Premenopausal	NN	O	O
patients	NN	O	O
with	NN	O	O
neither	NN	O	O
risk	NN	O	O
factor	NN	O	O
had	NN	O	O
a	NN	O	O
very	NN	O	O
low	NN	O	O
failure	NN	O	O
rate	NN	O	O
(	NN	O	O
2.6	NN	O	O
%	NN	O	O
at	NN	O	O
5	NN	O	O
years	NN	O	O
)	NN	O	O
,	NN	O	O
regardless	NN	O	O
of	NN	O	O
age	NN	O	O
.	NN	O	O

For	NN	O	O
postmenopausal	NN	O	O
patients	NN	O	O
risk	NN	O	O
of	NN	O	O
breast	NN	O	O
recurrence	NN	O	O
was	NN	O	O
determined	NN	O	O
both	NN	O	O
by	NN	O	O
adequacy	NN	O	O
of	NN	O	O
resection	NN	O	O
margins	NN	O	O
and	NN	O	O
grade	NN	O	O
,	NN	O	O
with	NN	O	O
a	NN	O	O
high	NN	O	O
local	NN	O	O
failure	NN	O	O
rate	NN	O	O
for	NN	O	O
patients	NN	O	O
having	NN	O	O
G3	NN	O	O
tumors	NN	O	O
with	NN	O	O
positive	NN	O	O
or	NN	O	O
indeterminate	NN	O	O
margins	NN	O	O
(	NN	O	O
31	NN	O	O
%	NN	O	O
at	NN	O	O
5	NN	O	O
years	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
authors	NN	O	O
conclude	NN	O	O
that	NN	O	O
the	NN	O	O
microscopic	NN	O	O
examination	NN	O	O
is	NN	O	O
the	NN	O	O
only	NN	O	O
useful	NN	O	O
tool	NN	O	O
for	NN	O	O
assessing	NN	O	O
the	NN	O	O
risk	NN	O	O
of	NN	O	O
local	NN	O	O
failure	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
quite	NN	O	O
low	NN	O	O
for	NN	O	O
the	NN	O	O
majority	NN	O	O
of	NN	O	O
patients	NN	O	O
treated	NN	O	O
with	NN	O	O
breast	NN	O	O
conservation	NN	O	O
.	NN	O	O

High-risk	NN	O	O
patients	NN	O	O
can	NN	O	O
be	NN	O	O
recognized	NN	O	O
morphologically	NN	O	O
.	NN	O	O

The	NN	O	O
age	NN	O	O
dependence	NN	O	O
of	NN	O	O
morphologic	NN	O	O
risk	NN	O	O
factors	NN	O	O
appears	NN	O	O
to	NN	O	O
explain	NN	O	O
the	NN	O	O
high	NN	O	O
local	NN	O	O
failure	NN	O	O
rate	NN	O	O
seen	NN	O	O
in	NN	O	O
patients	NN	O	O
younger	NN	O	O
than	NN	O	O
40	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tax	NN	O	B-protein
-independent	NN	O	O
binding	NN	O	O
of	NN	O	O
multiple	NN	O	O
cellular	NN	O	B-protein
factors	NN	O	I-protein
to	NN	O	O
Tax-response	NN	O	B-DNA
element	NN	O	I-DNA
DNA	NN	O	I-DNA
of	NN	O	O
HTLV-I	NN	O	O
.	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
I	NN	O	I-DNA
(	NN	O	I-DNA
HTLV-I	NN	O	I-DNA
)	NN	O	I-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
three	NN	O	O
copies	NN	O	O
of	NN	O	O
imperfect	NN	O	O
repeats	NN	O	O
of	NN	O	O
a	NN	O	O
21-base	NN	O	O
pair	NN	O	O
sequence	NN	O	O
designated	NN	O	O
here	NN	O	O
as	NN	O	O
TRE	NN	O	B-DNA
(	NN	O	O
Tax-response	NN	O	B-DNA
element	NN	O	I-DNA
)	NN	O	O
that	NN	O	O
is	NN	O	O
responsive	NN	O	O
to	NN	O	O
the	NN	O	O
virally	NN	O	B-protein
encoded	NN	O	I-protein
transactivator	NN	O	I-protein
protein	NN	O	I-protein
Tax	NN	O	B-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
and	NN	O	O
separated	NN	O	O
four	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
from	NN	O	O
C81-66-45	NN	O	O
cells	NN	O	O
,	NN	O	O
an	NN	O	O
HTLV-I	NN	O	B-cell_line
immortalized	NN	O	I-cell_line
Tax-expressing	NN	O	I-cell_line
human	NN	O	I-cell_line
T-lymphocyte	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
Salahuddin	NN	O	O
et	NN	O	O
al.	NN	O	O
,	NN	O	O
1983	NN	O	O
)	NN	O	O
,	NN	O	O
that	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
TRE-DNA	NN	O	B-DNA
,	NN	O	O
none	NN	O	O
of	NN	O	O
which	NN	O	O
are	NN	O	O
identical	NN	O	O
with	NN	O	O
the	NN	O	O
Tax-protein	NN	O	B-protein
.	NN	O	O

The	NN	O	O
proteins	NN	O	O
identified	NN	O	O
have	NN	O	O
molecular	NN	O	O
weights	NN	O	O
of	NN	O	O
about	NN	O	O
32	NN	O	O
,	NN	O	O
36	NN	O	O
to	NN	O	O
42	NN	O	O
,	NN	O	O
50	NN	O	O
and	NN	O	O
110	NN	O	O
kD	NN	O	O
.	NN	O	O

Four	NN	O	O
different	NN	O	O
methods	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
proteins	NN	O	O
.	NN	O	O

First	NN	O	O
,	NN	O	O
from	NN	O	O
different	NN	O	O
cell	NN	O	O
lines	NN	O	O
three	NN	O	O
or	NN	O	O
all	NN	O	O
four	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
specifically	NN	O	O
cross-linked	NN	O	O
by	NN	O	O
UV	NN	O	O
irradiation	NN	O	O
to	NN	O	O
the	NN	O	O
radioactively	NN	O	O
labeled	NN	O	O
TRE-DNA	NN	O	B-DNA
fragment	NN	O	I-DNA
.	NN	O	O

Second	NN	O	O
,	NN	O	O
TRE-DNA	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
sedimented	NN	O	O
through	NN	O	O
a	NN	O	O
glycerol	NN	O	O
density	NN	O	O
gradient	NN	O	O
at	NN	O	O
rates	NN	O	O
corresponding	NN	O	O
to	NN	O	O
proteins	NN	O	O
of	NN	O	O
native	NN	O	O
molecular	NN	O	O
weights	NN	O	O
of	NN	O	O
35	NN	O	O
to	NN	O	O
50	NN	O	O
kD	NN	O	O
and	NN	O	O
110	NN	O	O
kD	NN	O	O
.	NN	O	O

Third	NN	O	O
,	NN	O	O
only	NN	O	O
the	NN	O	O
50	NN	O	B-protein
kD	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
retained	NN	O	O
on	NN	O	O
a	NN	O	O
biotinylated	NN	O	O
DNA-streptavidin	NN	O	O
matrix	NN	O	O
when	NN	O	O
the	NN	O	O
DNA	NN	O	O
fragment	NN	O	O
contained	NN	O	O
the	NN	O	O
TRE-DNA	NN	O	B-DNA
.	NN	O	O

Fourth	NN	O	O
,	NN	O	O
extensive	NN	O	O
purification	NN	O	O
by	NN	O	O
several	NN	O	O
cycles	NN	O	O
of	NN	O	O
TRE	NN	O	B-DNA
-DNA	NN	O	O
affinity	NN	O	O
chromatography	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
32	NN	O	B-protein
,	NN	O	I-protein
36	NN	O	I-protein
to	NN	O	I-protein
42	NN	O	I-protein
and	NN	O	I-protein
110	NN	O	I-protein
kD	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
to	NN	O	O
less	NN	O	O
extent	NN	O	O
the	NN	O	O
50	NN	O	B-protein
kD	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

Two	NN	O	O
abundant	NN	O	O
proteins	NN	O	O
of	NN	O	O
75	NN	O	O
and	NN	O	O
80	NN	O	O
kD	NN	O	O
were	NN	O	O
competed	NN	O	O
out	NN	O	O
by	NN	O	O
poly	NN	O	O
[	NN	O	O
d	NN	O	O
(	NN	O	O
I-C	NN	O	O
)	NN	O	O
]	NN	O	O
in	NN	O	O
all	NN	O	O
reactions	NN	O	O
.	NN	O	O

The	NN	O	O
cAMP-response	NN	O	B-DNA
element	NN	O	I-DNA
CRE	NN	O	B-DNA
,	NN	O	O
TGACGTCA	NN	O	O
,	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
21	NN	O	B-DNA
base-pair	NN	O	I-DNA
sequence	NN	O	I-DNA
,	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
essential	NN	O	O
for	NN	O	O
specific	NN	O	O
protein-	NN	O	O
TRE	NN	O	B-DNA
-DNA	NN	O	O
interactions	NN	O	O
because	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
G	NN	O	O
's	NN	O	O
destroys	NN	O	O
this	NN	O	O
complex	NN	O	O
.	NN	O	O

This	NN	O	O
result	NN	O	O
suggests	NN	O	O
that	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
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
protein-TRE-DNA	NN	O	B-protein
complex	NN	O	I-protein
and	NN	O	O
in	NN	O	O
mediating	NN	O	O
the	NN	O	O
Tax	NN	O	B-protein
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

Two	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
the	NN	O	O
immunoglobulin	NN	O	B-DNA
enhancer	NN	O	I-DNA
microE5/kappa	NN	O	B-DNA
2	NN	O	I-DNA
motif	NN	O	I-DNA
.	NN	O	O

Activity	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
heavy	NN	O	I-DNA
and	NN	O	I-DNA
kappa	NN	O	I-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
gene	NN	O	I-DNA
enhancers	NN	O	I-DNA
depends	NN	O	O
on	NN	O	O
a	NN	O	O
complex	NN	O	O
interplay	NN	O	O
of	NN	O	O
ubiquitous	NN	O	B-protein
and	NN	O	I-protein
developmentally	NN	O	I-protein
regulated	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

Two	NN	O	O
complementary	NN	O	B-DNA
DNAs	NN	O	I-DNA
were	NN	O	O
isolated	NN	O	O
that	NN	O	O
encode	NN	O	O
proteins	NN	O	O
,	NN	O	O
denoted	NN	O	O
ITF-1	NN	O	B-protein
and	NN	O	O
ITF-2	NN	O	B-protein
,	NN	O	O
that	NN	O	O
are	NN	O	O
expressed	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
and	NN	O	O
bind	NN	O	O
the	NN	O	O
microE5/kappa	NN	O	B-DNA
2	NN	O	I-DNA
motif	NN	O	I-DNA
found	NN	O	O
in	NN	O	O
both	NN	O	O
heavy	NN	O	B-DNA
and	NN	O	I-DNA
kappa	NN	O	I-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
enhancers	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
complementary	NN	O	B-DNA
DNAs	NN	O	I-DNA
are	NN	O	O
the	NN	O	O
products	NN	O	O
of	NN	O	O
distinct	NN	O	O
genes	NN	O	O
,	NN	O	O
yet	NN	O	O
both	NN	O	O
ITF-1	NN	O	B-protein
and	NN	O	O
ITF-2	NN	O	B-protein
are	NN	O	O
structurally	NN	O	O
and	NN	O	O
functionally	NN	O	O
similar	NN	O	O
.	NN	O	O

The	NN	O	O
two	NN	O	O
proteins	NN	O	O
interact	NN	O	O
with	NN	O	O
one	NN	O	O
another	NN	O	O
through	NN	O	O
their	NN	O	O
putative	NN	O	O
helix-loop-helix	NN	O	B-protein
motifs	NN	O	I-protein
and	NN	O	O
each	NN	O	O
possesses	NN	O	O
a	NN	O	O
distinct	NN	O	O
domain	NN	O	O
that	NN	O	O
dictates	NN	O	O
transcription	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Elevated	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
concentrations	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
glucocorticoid	NN	O	O
therapy	NN	O	O
in	NN	O	O
peripheral	NN	O	B-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
atopic	NN	O	O
dermatitis	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
glucocorticoid	NN	O	O
binding	NN	O	O
sites	NN	O	O
in	NN	O	O
peripheral	NN	O	B-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
atopic	NN	O	O
dermatitis	NN	O	O
(	NN	O	O
AD	NN	O	O
)	NN	O	O
and	NN	O	O
healthy	NN	O	O
controls	NN	O	O
were	NN	O	O
determined	NN	O	O
under	NN	O	O
baseline	NN	O	O
conditions	NN	O	O
and	NN	O	O
after	NN	O	O
a	NN	O	O
defined	NN	O	O
oral	NN	O	O
glucocorticoid	NN	O	O
treatment	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
AD	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
15	NN	O	O
)	NN	O	O
exhibited	NN	O	O
significantly	NN	O	O
more	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GR	NN	O	B-protein
)	NN	O	O
per	NN	O	O
cell	NN	O	O
than	NN	O	O
the	NN	O	O
control	NN	O	O
group	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
22	NN	O	O
)	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
GR	NN	O	B-protein
affinity	NN	O	O
did	NN	O	O
not	NN	O	O
differ	NN	O	O
.	NN	O	O

Methylprednisolone	NN	O	O
treatment	NN	O	O
resulted	NN	O	O
in	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-protein
sites	NN	O	O
per	NN	O	O
cell	NN	O	O
in	NN	O	O
the	NN	O	O
steroid-treated	NN	O	O
control	NN	O	O
group	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
10	NN	O	O
)	NN	O	O
in	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
patients	NN	O	O
.	NN	O	O

The	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
was	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
methylprednisolone	NN	O	O
treatment	NN	O	O
in	NN	O	O
either	NN	O	O
group	NN	O	O
.	NN	O	O

In	NN	O	O
view	NN	O	O
of	NN	O	O
the	NN	O	O
therapeutic	NN	O	O
efficiency	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
AD	NN	O	O
and	NN	O	O
findings	NN	O	O
of	NN	O	O
abnormal	NN	O	O
cAMP	NN	O	O
and	NN	O	O
cAMP-phosphodiesterase	NN	O	O
activity	NN	O	O
,	NN	O	O
the	NN	O	O
elevated	NN	O	O
GR	NN	O	B-protein
concentrations	NN	O	O
in	NN	O	O
AD	NN	O	O
lend	NN	O	O
support	NN	O	O
to	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
of	NN	O	O
a	NN	O	O
compensatory	NN	O	O
GR	NN	O	B-protein
upregulation	NN	O	O
due	NN	O	O
to	NN	O	O
an	NN	O	O
insufficient	NN	O	O
action	NN	O	O
of	NN	O	O
endogenous	NN	O	O
cortisol	NN	O	O
or	NN	O	O
to	NN	O	O
altered	NN	O	O
cAMP-induced	NN	O	O
GR	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
actions	NN	O	O
of	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
suggest	NN	O	O
a	NN	O	O
novel	NN	O	O
step	NN	O	O
in	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

Cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
are	NN	O	O
immunosuppressive	NN	O	O
compounds	NN	O	O
that	NN	O	O
have	NN	O	O
similar	NN	O	O
inhibitory	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
lymphokines	NN	O	B-protein
produced	NN	O	O
by	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Despite	NN	O	O
their	NN	O	O
similar	NN	O	O
effects	NN	O	O
the	NN	O	O
drugs	NN	O	O
bind	NN	O	O
to	NN	O	O
two	NN	O	O
different	NN	O	O
cytosolic	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
cyclophilin	NN	O	B-protein
and	NN	O	O
FKBP	NN	O	B-protein
respectively	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
they	NN	O	O
have	NN	O	O
different	NN	O	O
modes	NN	O	O
of	NN	O	O
action	NN	O	O
.	NN	O	O

Using	NN	O	O
constructs	NN	O	O
in	NN	O	O
which	NN	O	O
mRNA	NN	O	B-RNA
production	NN	O	O
controlled	NN	O	O
by	NN	O	O
a	NN	O	O
specific	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
could	NN	O	O
be	NN	O	O
readily	NN	O	O
measured	NN	O	O
we	NN	O	O
found	NN	O	O
that	NN	O	O
both	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
completely	NN	O	O
inhibited	NN	O	O
transcription	NN	O	O
activated	NN	O	O
by	NN	O	O
NF-AT	NN	O	B-protein
,	NN	O	O
NFIL2	NN	O	B-protein
A	NN	O	I-protein
,	NN	O	O
NFIL2	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
partially	NN	O	O
inhibited	NN	O	O
transcription	NN	O	O
activated	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

Cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
inhibited	NN	O	O
only	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
that	NN	O	O
was	NN	O	O
dependent	NN	O	O
on	NN	O	O
Ca2+	NN	O	O
mobilization	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
did	NN	O	O
not	NN	O	O
inhibit	NN	O	O
Ca2+	NN	O	O
mobilization	NN	O	O
dependent	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-RNA
mRNA	NN	O	I-RNA
indicating	NN	O	O
that	NN	O	O
only	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
signalling	NN	O	O
pathways	NN	O	O
regulated	NN	O	O
by	NN	O	O
Ca2+	NN	O	O
is	NN	O	O
sensitive	NN	O	O
to	NN	O	O
these	NN	O	O
drugs	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
did	NN	O	O
not	NN	O	O
observe	NN	O	O
any	NN	O	O
qualitative	NN	O	O
differences	NN	O	O
between	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
FK506	NN	O	O
on	NN	O	O
six	NN	O	O
different	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
which	NN	O	O
suggests	NN	O	O
that	NN	O	O
these	NN	O	O
drugs	NN	O	O
may	NN	O	O
interfere	NN	O	O
with	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
Ca2+	NN	O	O
dependent	NN	O	O
step	NN	O	O
that	NN	O	O
regulates	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
internal	NN	O	O
methionine	NN	O	O
codons	NN	O	O
of	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
II	NN	O	I-DNA
rex	NN	O	I-DNA
gene	NN	O	I-DNA
are	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
p24rex	NN	O	B-protein
production	NN	O	O
or	NN	O	O
virus	NN	O	O
replication	NN	O	O
and	NN	O	O
transformation	NN	O	O
.	NN	O	O

Human	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
types	NN	O	O
I	NN	O	O
(	NN	O	O
HTLV-I	NN	O	O
)	NN	O	O
and	NN	O	O
II	NN	O	O
(	NN	O	O
HTLV-II	NN	O	O
)	NN	O	O
have	NN	O	O
two	NN	O	O
nonstructural	NN	O	B-DNA
trans-acting	NN	O	I-DNA
regulatory	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
tax	NN	O	B-DNA
and	NN	O	O
rex	NN	O	B-DNA
,	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
region	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
viral	NN	O	B-DNA
genome	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
tax	NN	O	B-protein
gene	NN	O	I-protein
product	NN	O	I-protein
(	NN	O	O
HTLV-I	NN	O	B-protein
p40tax	NN	O	I-protein
and	NN	O	O
HTLV-II	NN	O	B-protein
p37tax	NN	O	I-protein
)	NN	O	O
is	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activator	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
.	NN	O	O

The	NN	O	O
rex	NN	O	B-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
two	NN	O	O
protein	NN	O	O
products	NN	O	O
,	NN	O	O
p27rex/p21rex	NN	O	B-protein
and	NN	O	O
p26rex/p24rex	NN	O	B-protein
in	NN	O	O
HTLV-I	NN	O	O
and	NN	O	O
HTLV-II	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Rex	NN	O	B-protein
acts	NN	O	O
posttranscriptionally	NN	O	O
to	NN	O	O
facilitate	NN	O	O
accumulation	NN	O	O
of	NN	O	O
full-length	NN	O	O
gag/pol	NN	O	B-protein
and	NN	O	O
singly	NN	O	O
spliced	NN	O	O
env	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
of	NN	O	O
HTLV-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Previous	NN	O	O
studies	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
first	NN	O	O
ATG	NN	O	O
of	NN	O	O
the	NN	O	O
rex	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
critical	NN	O	O
for	NN	O	O
Rex	NN	O	B-protein
production	NN	O	O
and	NN	O	O
function	NN	O	O
.	NN	O	O

The	NN	O	O
importance	NN	O	O
of	NN	O	O
the	NN	O	O
internal	NN	O	O
ATGs	NN	O	O
to	NN	O	O
Rex	NN	O	B-protein
function	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
in	NN	O	O
vitro	NN	O	O
mutagenesis	NN	O	O
of	NN	O	O
the	NN	O	O
HTLV-I	NN	O	O
rex	NN	O	B-DNA
gene	NN	O	I-DNA
has	NN	O	O
provided	NN	O	O
indirect	NN	O	O
evidence	NN	O	O
which	NN	O	O
suggests	NN	O	O
that	NN	O	O
p21rex	NN	O	B-protein
,	NN	O	O
and	NN	O	O
by	NN	O	O
analogy	NN	O	O
HTLV-II	NN	O	O
p24rex	NN	O	B-protein
,	NN	O	O
results	NN	O	O
from	NN	O	O
initiation	NN	O	O
at	NN	O	O
an	NN	O	O
internal	NN	O	O
AUG	NN	O	O
of	NN	O	O
the	NN	O	O
tax/rex	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

By	NN	O	O
using	NN	O	O
an	NN	O	O
infectious	NN	O	O
molecular	NN	O	O
clone	NN	O	O
of	NN	O	O
HTLV-II	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
the	NN	O	O
internal	NN	O	O
ATGs	NN	O	O
of	NN	O	O
the	NN	O	O
rex	NN	O	B-DNA
gene	NN	O	I-DNA
on	NN	O	O
Rex	NN	O	B-protein
protein	NN	O	O
production	NN	O	O
and	NN	O	O
function	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
p24rex	NN	O	B-protein
of	NN	O	O
HTLV-II	NN	O	O
is	NN	O	O
not	NN	O	O
initiated	NN	O	O
at	NN	O	O
an	NN	O	O
internal	NN	O	O
AUG	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
internal	NN	O	O
methionine	NN	O	O
codons	NN	O	O
are	NN	O	O
not	NN	O	O
crucial	NN	O	O
to	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
rex	NN	O	B-DNA
gene	NN	O	I-DNA
and	NN	O	O
,	NN	O	O
ultimately	NN	O	O
,	NN	O	O
the	NN	O	O
transforming	NN	O	O
properties	NN	O	O
of	NN	O	O
the	NN	O	O
virus	NN	O	O
.	NN	O	O

-DOCSTART-	O

Astrocytes	NN	O	B-cell_type
and	NN	O	O
glioblastoma	NN	O	B-cell_type
cells	NN	O	I-cell_type
express	NN	O	O
novel	NN	O	O
octamer-DNA	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
ubiquitous	NN	O	B-protein
Oct-1	NN	O	I-protein
and	NN	O	I-protein
B	NN	O	I-protein
cell	NN	O	I-protein
type	NN	O	I-protein
Oct-2	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
'octamer	NN	O	B-DNA
'	NN	O	I-DNA
sequence	NN	O	I-DNA
,	NN	O	O
ATGCAAAT	NN	O	O
or	NN	O	O
its	NN	O	O
complement	NN	O	O
ATTTGCAT	NN	O	O
,	NN	O	O
is	NN	O	O
a	NN	O	O
key	NN	O	O
element	NN	O	O
for	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
B-lymphocytes	NN	O	B-cell_type
as	NN	O	O
well	NN	O	O
as	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
housekeeping	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
all	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

In	NN	O	O
lymphocytes	NN	O	B-cell_type
,	NN	O	O
the	NN	O	O
octamer-binding	NN	O	B-protein
protein	NN	O	I-protein
Oct-2A	NN	O	B-protein
and	NN	O	O
variants	NN	O	O
thereof	NN	O	O
are	NN	O	O
thought	NN	O	O
to	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
B-cell	NN	O	B-DNA
specific	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
,	NN	O	O
while	NN	O	O
the	NN	O	O
ubiquitous	NN	O	O
protein	NN	O	O
Oct-1	NN	O	O
seems	NN	O	O
to	NN	O	O
control	NN	O	O
general	NN	O	O
octamer	NN	O	B-DNA
site	NN	O	I-DNA
-dependent	NN	O	O
transcription	NN	O	O
.	NN	O	O

Various	NN	O	O
other	NN	O	O
genes	NN	O	O
,	NN	O	O
for	NN	O	O
example	NN	O	O
interleukin-1	NN	O	B-DNA
and	NN	O	I-DNA
MHC	NN	O	I-DNA
class	NN	O	I-DNA
II	NN	O	I-DNA
genes	NN	O	I-DNA
,	NN	O	O
contain	NN	O	O
an	NN	O	O
octamer	NN	O	B-DNA
sequence	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
and	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
cells	NN	O	O
of	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

This	NN	O	O
prompted	NN	O	O
us	NN	O	O
to	NN	O	O
analyze	NN	O	O
the	NN	O	O
octamer-binding	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
the	NN	O	O
latter	NN	O	O
cells	NN	O	O
.	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
at	NN	O	O
least	NN	O	O
six	NN	O	O
novel	NN	O	O
octamer	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
cultured	NN	O	B-cell_type
mouse	NN	O	I-cell_type
astrocytes	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
proteins	NN	O	O
are	NN	O	O
differentially	NN	O	O
expressed	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
glioblastoma	NN	O	I-cell_line
and	NN	O	I-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
nervous	NN	O	B-protein
system-derived	NN	O	I-protein
(	NN	O	I-protein
N-Oct	NN	O	I-protein
)	NN	O	I-protein
proteins	NN	O	I-protein
bound	NN	O	O
to	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
DNA	NN	O	I-DNA
sequence	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
manner	NN	O	O
which	NN	O	O
is	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
the	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2A	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
relationship	NN	O	O
of	NN	O	O
the	NN	O	O
N-Oct	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2A	NN	O	B-protein
was	NN	O	O
analyzed	NN	O	O
by	NN	O	O
proteolytic	NN	O	O
clipping	NN	O	O
bandshift	NN	O	O
assays	NN	O	O
and	NN	O	O
by	NN	O	O
their	NN	O	O
reactivity	NN	O	O
towards	NN	O	O
antisera	NN	O	O
raised	NN	O	O
against	NN	O	O
recombinant	NN	O	O
Oct-1	NN	O	B-protein
and	NN	O	O
Oct-2A	NN	O	B-protein
proteins	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
assays	NN	O	O
,	NN	O	O
all	NN	O	O
N-Oct-factors	NN	O	B-protein
were	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
distinct	NN	O	O
from	NN	O	O
the	NN	O	O
ubiquitous	NN	O	B-protein
Oct-1	NN	O	I-protein
and	NN	O	O
the	NN	O	O
lymphoid-specific	NN	O	B-protein
Oct-2A	NN	O	I-protein
proteins	NN	O	O
.	NN	O	O

In	NN	O	O
melanoma	NN	O	B-cell_type
cells	NN	O	I-cell_type
that	NN	O	O
contain	NN	O	O
the	NN	O	O
N-Oct-3	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
a	NN	O	O
transfected	NN	O	O
lymphocyte-specific	NN	O	O
promoter	NN	O	O
was	NN	O	O
neither	NN	O	O
activated	NN	O	O
nor	NN	O	O
was	NN	O	O
it	NN	O	O
repressed	NN	O	O
upon	NN	O	O
contransfection	NN	O	O
with	NN	O	O
an	NN	O	O
Oct-2A	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
speculate	NN	O	O
that	NN	O	O
N-Oct-3	NN	O	B-protein
and	NN	O	O
other	NN	O	O
N-Oct	NN	O	B-protein
factors	NN	O	I-protein
have	NN	O	O
a	NN	O	O
specific	NN	O	O
role	NN	O	O
in	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
nervous	NN	O	O
system	NN	O	O
.	NN	O	O

-DOCSTART-	O

Detection	NN	O	O
in	NN	O	O
non-erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
a	NN	O	O
factor	NN	O	O
with	NN	O	O
the	NN	O	O
binding	NN	O	O
characteristics	NN	O	O
of	NN	O	O
the	NN	O	O
erythroid	NN	O	B-protein
cell	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
EF1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
erythroid	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
erythroid	NN	O	B-protein
factor-1	NN	O	I-protein
(	NN	O	O
EF1	NN	O	B-protein
)	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
transcription	NN	O	O
of	NN	O	O
erythroid-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
factor	NN	O	O
with	NN	O	O
the	NN	O	O
mobility	NN	O	O
and	NN	O	O
sequence-specific	NN	O	O
DNA-binding	NN	O	O
characteristics	NN	O	O
of	NN	O	O
EF1	NN	O	B-protein
at	NN	O	O
low	NN	O	O
abundance	NN	O	O
in	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
non-erythroid	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
.	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
EF1	NN	O	B-protein
-like	NN	O	O
activity	NN	O	O
in	NN	O	O
non-erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
indicates	NN	O	O
that	NN	O	O
this	NN	O	O
factor	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	B-DNA
expressed	NN	O	O
in	NN	O	O
such	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Protein	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
H-7	NN	O	O
blocks	NN	O	O
accumulation	NN	O	O
of	NN	O	O
unspliced	NN	O	B-RNA
mRNA	NN	O	I-RNA
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
.	NN	O	O

Rex	NN	O	B-protein
,	NN	O	O
the	NN	O	O
post-transcriptional	NN	O	B-protein
regulator	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-I	NN	O	O
)	NN	O	O
,	NN	O	O
is	NN	O	O
known	NN	O	O
to	NN	O	O
induce	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
unspliced	NN	O	B-RNA
viral	NN	O	I-RNA
gag-pol	NN	O	I-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Rex	NN	O	B-protein
is	NN	O	O
a	NN	O	O
phosphoprotein	NN	O	B-protein
found	NN	O	O
in	NN	O	O
the	NN	O	O
cell	NN	O	O
nucleolus	NN	O	O
,	NN	O	O
whose	NN	O	O
function	NN	O	O
may	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
its	NN	O	O
localization	NN	O	O
and	NN	O	O
phosphorylation	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
phosphorylation	NN	O	O
on	NN	O	O
Rex	NN	O	B-protein
function	NN	O	O
by	NN	O	O
using	NN	O	O
a	NN	O	O
protein	NN	O	O
kinase	NN	O	O
inhibitor	NN	O	O
,	NN	O	O
H-7	NN	O	O
[	NN	O	O
1-	NN	O	O
(	NN	O	O
5-isoquinolinyl-sulfonyl	NN	O	O
)	NN	O	O
-2-methylpiperazine	NN	O	O
]	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
an	NN	O	O
HTLV-I	NN	O	B-cell_line
infected	NN	O	I-cell_line
human	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
with	NN	O	O
H-7	NN	O	O
blocked	NN	O	O
specifically	NN	O	O
accumulation	NN	O	O
of	NN	O	O
the	NN	O	O
unspliced	NN	O	B-RNA
gag-pol	NN	O	I-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
decreased	NN	O	O
Gag	NN	O	B-protein
protein	NN	O	I-protein
synthesis	NN	O	O
that	NN	O	O
corresponds	NN	O	O
with	NN	O	O
the	NN	O	O
decreased	NN	O	O
in	NN	O	O
vivo	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Rex	NN	O	B-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
other	NN	O	O
viral	NN	O	O
and	NN	O	O
cellular	NN	O	O
products	NN	O	O
have	NN	O	O
not	NN	O	O
been	NN	O	O
influenced	NN	O	O
by	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
H-7	NN	O	O
used	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Rex	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
viral	NN	O	O
RNA	NN	O	O
partition	NN	O	O
of	NN	O	O
HTLV-I	NN	O	O
.	NN	O	O

-DOCSTART-	O

Increased	NN	O	O
glucocorticoid	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
CD4+	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
clonal	NN	O	I-cell_line
lines	NN	O	I-cell_line
grown	NN	O	O
in	NN	O	O
serum-free	NN	O	O
media	NN	O	O
.	NN	O	O

CEM-C7	NN	O	B-cell_line
,	NN	O	O
a	NN	O	O
human	NN	O	B-cell_line
leukemic	NN	O	I-cell_line
CD4+	NN	O	I-cell_line
T-lymphocyte	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
three	NN	O	O
of	NN	O	O
its	NN	O	O
subclones	NN	O	O
,	NN	O	O
CEM-4R4	NN	O	B-cell_line
,	NN	O	O
CEM-3R43	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
ICR-27	NN	O	B-cell_line
,	NN	O	O
previously	NN	O	O
cultured	NN	O	O
in	NN	O	O
a	NN	O	O
medium	NN	O	O
supplemented	NN	O	O
with	NN	O	O
5	NN	O	O
to	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
have	NN	O	O
been	NN	O	O
adapted	NN	O	O
to	NN	O	O
serum-free	NN	O	O
media	NN	O	O
.	NN	O	O

The	NN	O	O
best	NN	O	O
medium	NN	O	O
of	NN	O	O
those	NN	O	O
tested	NN	O	O
was	NN	O	O
RPMI	NN	O	O
1640	NN	O	O
supplemented	NN	O	O
with	NN	O	O
5	NN	O	O
micrograms/ml	NN	O	O
each	NN	O	O
transferrin	NN	O	B-protein
and	NN	O	O
insulin	NN	O	B-protein
+	NN	O	O
5	NN	O	O
ng/ml	NN	O	O
sodium	NN	O	O
selenite	NN	O	O
+/-	NN	O	O
0.1	NN	O	O
%	NN	O	O
bovine	NN	O	B-protein
serum	NN	O	I-protein
albumin	NN	O	I-protein
.	NN	O	O

While	NN	O	O
growing	NN	O	O
either	NN	O	O
with	NN	O	O
or	NN	O	O
without	NN	O	O
albumin	NN	O	O
,	NN	O	O
the	NN	O	O
several	NN	O	O
clonal	NN	O	B-cell_line
lines	NN	O	I-cell_line
of	NN	O	O
CEM	NN	O	B-cell_line
cells	NN	O	O
displayed	NN	O	O
growth	NN	O	O
similar	NN	O	O
to	NN	O	O
serum-supplemented	NN	O	O
cultures	NN	O	O
.	NN	O	O

Cell	NN	O	O
proliferation	NN	O	O
of	NN	O	O
CEM-C7	NN	O	B-cell_line
cells	NN	O	O
cultured	NN	O	O
in	NN	O	O
both	NN	O	O
serum-free	NN	O	O
media	NN	O	O
has	NN	O	O
been	NN	O	O
sustained	NN	O	O
for	NN	O	O
3	NN	O	O
mo	NN	O	O
.	NN	O	O

with	NN	O	O
culture	NN	O	O
doubling	NN	O	O
times	NN	O	O
of	NN	O	O
about	NN	O	O
25	NN	O	O
h	NN	O	O
for	NN	O	O
both	NN	O	O
serum-supplemented	NN	O	B-cell_line
and	NN	O	I-cell_line
serum-free	NN	O	I-cell_line
cultures	NN	O	I-cell_line
(	NN	O	O
viability	NN	O	O
greater	NN	O	O
than	NN	O	O
or	NN	O	O
equal	NN	O	O
to	NN	O	O
90	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

Cell	NN	O	O
morphology	NN	O	O
remained	NN	O	O
essentially	NN	O	O
the	NN	O	O
same	NN	O	O
in	NN	O	O
serum-free	NN	O	O
or	NN	O	O
serum	NN	O	O
containing	NN	O	O
media	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
CD4	NN	O	B-protein
,	NN	O	O
a	NN	O	O
marker	NN	O	O
for	NN	O	O
T-derived	NN	O	B-cell_type
lymphoid	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
was	NN	O	O
not	NN	O	O
significantly	NN	O	O
different	NN	O	O
in	NN	O	O
serum-free	NN	O	O
medium	NN	O	O
.	NN	O	O

When	NN	O	O
grown	NN	O	O
in	NN	O	O
serum-free	NN	O	O
medium	NN	O	O
,	NN	O	O
CEM-C7	NN	O	B-cell_line
cells	NN	O	O
exhibited	NN	O	O
increased	NN	O	O
steroid	NN	O	O
responsiveness	NN	O	O
as	NN	O	O
evidenced	NN	O	O
by	NN	O	O
increased	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
sites	NN	O	O
,	NN	O	O
increased	NN	O	O
induction	NN	O	O
of	NN	O	O
glutamine	NN	O	B-protein
synthetase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
cell	NN	O	O
lysis	NN	O	O
at	NN	O	O
lower	NN	O	O
concentrations	NN	O	O
of	NN	O	O
steroid	NN	O	O
.	NN	O	O

Receptor	NN	O	O
mutant	NN	O	O
subclones	NN	O	O
of	NN	O	O
CEM-C7	NN	O	B-cell_line
,	NN	O	O
which	NN	O	O
are	NN	O	O
proven	NN	O	O
to	NN	O	O
be	NN	O	O
completely	NN	O	O
unresponsive	NN	O	O
to	NN	O	O
micromolar	NN	O	O
concentrations	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
when	NN	O	O
grown	NN	O	O
in	NN	O	O
serum-supplemented	NN	O	O
medium	NN	O	O
,	NN	O	O
become	NN	O	O
partially	NN	O	O
sensitive	NN	O	O
to	NN	O	O
the	NN	O	O
hormone	NN	O	O
after	NN	O	O
growth	NN	O	O
in	NN	O	O
defined	NN	O	O
medium	NN	O	O
.	NN	O	O

The	NN	O	O
increased	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
CEM-C7	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
its	NN	O	O
subclones	NN	O	O
to	NN	O	O
dexamethasone	NN	O	O
in	NN	O	O
serum-free	NN	O	O
medium	NN	O	O
returned	NN	O	O
to	NN	O	O
previous	NN	O	O
levels	NN	O	O
when	NN	O	O
these	NN	O	O
cells	NN	O	O
were	NN	O	O
recultured	NN	O	O
in	NN	O	O
serum-containing	NN	O	O
medium	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
substances	NN	O	O
in	NN	O	O
serum	NN	O	O
influence	NN	O	O
steroid	NN	O	O
effects	NN	O	O
on	NN	O	O
these	NN	O	O
cells	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
molecular	NN	O	O
details	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	O
hormone	NN	O	O
action	NN	O	O
may	NN	O	O
be	NN	O	O
pursued	NN	O	O
more	NN	O	O
precisely	NN	O	O
in	NN	O	O
a	NN	O	O
clearly	NN	O	O
defined	NN	O	O
culture	NN	O	O
medium	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
on	NN	O	O
human	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
mononuclear	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
changes	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
yang-deficiency	NN	O	O
]	NN	O	O

It	NN	O	O
was	NN	O	O
found	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
former	NN	O	O
works	NN	O	O
,	NN	O	O
the	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
(	NN	O	O
GCR	NN	O	B-protein
)	NN	O	O
on	NN	O	O
peripheral	NN	O	B-cell_type
mixed	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
Yang-deficiency	NN	O	O
were	NN	O	O
decreased	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
work	NN	O	O
,	NN	O	O
the	NN	O	O
mixed	NN	O	B-cell_type
leucocytes	NN	O	I-cell_type
were	NN	O	O
further	NN	O	O
separated	NN	O	O
into	NN	O	O
mononuclear	NN	O	B-cell_type
(	NN	O	I-cell_type
MNL	NN	O	I-cell_type
)	NN	O	I-cell_type
and	NN	O	I-cell_type
polymorphonuclear	NN	O	I-cell_type
(	NN	O	I-cell_type
PML	NN	O	I-cell_type
)	NN	O	I-cell_type
leucocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
GCR	NN	O	B-protein
were	NN	O	O
determined	NN	O	O
in	NN	O	O
each	NN	O	O
part	NN	O	O
of	NN	O	O
leucocytes	NN	O	B-cell_type
.	NN	O	O

GCR	NN	O	B-protein
on	NN	O	O
MNL	NN	O	B-cell_type
and	NN	O	O
PML	NN	O	B-cell_type
in	NN	O	O
6	NN	O	O
Yang	NN	O	O
deficient	NN	O	O
patients	NN	O	O
were	NN	O	O
3473	NN	O	O
+/-	NN	O	O
413	NN	O	O
and	NN	O	O
4433	NN	O	O
+/-	NN	O	O
651	NN	O	O
sites/cell	NN	O	O
respectively	NN	O	O
,	NN	O	O
statistically	NN	O	O
significant	NN	O	O
from	NN	O	O
the	NN	O	O
normal	NN	O	O
control	NN	O	O
group	NN	O	O
(	NN	O	O
4462	NN	O	O
+/-	NN	O	O
962	NN	O	O
and	NN	O	O
5622	NN	O	O
+/-	NN	O	O
782	NN	O	O
sites/cell	NN	O	O
respectively	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

GCR	NN	O	B-protein
on	NN	O	O
MNL	NN	O	B-cell_type
,	NN	O	O
PML	NN	O	B-cell_type
and	NN	O	O
mixed	NN	O	B-cell_type
leucocytes	NN	O	I-cell_type
in	NN	O	O
5	NN	O	O
patients	NN	O	O
were	NN	O	O
determined	NN	O	O
simultaneously	NN	O	O
,	NN	O	O
and	NN	O	O
all	NN	O	O
lowered	NN	O	O
from	NN	O	O
the	NN	O	O
control	NN	O	O
group	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
were	NN	O	O
3369	NN	O	O
+/-	NN	O	O
370	NN	O	O
,	NN	O	O
4986	NN	O	O
+/-	NN	O	O
419	NN	O	O
and	NN	O	O
4524	NN	O	O
+/-	NN	O	O
852	NN	O	O
sites/cell	NN	O	O
respectively	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
lowest	NN	O	O
GCR	NN	O	B-protein
on	NN	O	O
MNL	NN	O	B-cell_type
and	NN	O	O
highest	NN	O	O
on	NN	O	O
PML	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
ubiquitous	NN	O	B-protein
octamer-binding	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
is	NN	O	O
sufficient	NN	O	O
for	NN	O	O
transcription	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

All	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
contain	NN	O	O
a	NN	O	O
conserved	NN	O	O
octanucleotide	NN	O	B-DNA
promoter	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
ATGCAAAT	NN	O	O
,	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
required	NN	O	O
for	NN	O	O
their	NN	O	O
normal	NN	O	O
B-cell-specific	NN	O	O
transcription	NN	O	O
.	NN	O	O

Proteins	NN	O	O
that	NN	O	O
bind	NN	O	O
this	NN	O	O
octamer	NN	O	O
have	NN	O	O
been	NN	O	O
purified	NN	O	O
,	NN	O	O
and	NN	O	O
cDNAs	NN	O	B-DNA
encoding	NN	O	O
octamer-binding	NN	O	B-protein
proteins	NN	O	I-protein
have	NN	O	O
been	NN	O	O
cloned	NN	O	O
.	NN	O	O

Some	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
(	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
OTF-2	NN	O	B-protein
)	NN	O	O
are	NN	O	O
lymphoid	NN	O	O
specific	NN	O	O
,	NN	O	O
whereas	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
other	NN	O	O
,	NN	O	O
and	NN	O	O
possibly	NN	O	O
more	NN	O	O
(	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
OTF-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
is	NN	O	O
found	NN	O	O
ubiquitously	NN	O	O
in	NN	O	O
all	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

The	NN	O	O
exact	NN	O	O
role	NN	O	O
of	NN	O	O
these	NN	O	O
different	NN	O	O
proteins	NN	O	O
in	NN	O	O
directing	NN	O	O
the	NN	O	O
tissue-specific	NN	O	O
expression	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
unclear	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
two	NN	O	O
human	NN	O	B-cell_line
pre-B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
contain	NN	O	O
extremely	NN	O	O
low	NN	O	O
levels	NN	O	O
of	NN	O	O
OTF-2	NN	O	B-protein
yet	NN	O	O
still	NN	O	O
express	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
steady-state	NN	O	B-RNA
immunoglobulin	NN	O	I-RNA
heavy-chain	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
vivo	NN	O	O
and	NN	O	O
efficiently	NN	O	O
transcribe	NN	O	O
an	NN	O	O
immunoglobulin	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
a	NN	O	O
highly	NN	O	O
enriched	NN	O	O
preparation	NN	O	O
of	NN	O	O
OTF-1	NN	O	B-protein
made	NN	O	O
from	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
pre-B	NN	O	B-cell_line
cells	NN	O	I-cell_line
or	NN	O	O
from	NN	O	O
HeLa	NN	O	B-cell_line
cells	NN	O	I-cell_line
specifically	NN	O	O
stimulated	NN	O	O
in	NN	O	O
vitro	NN	O	O
transcription	NN	O	O
of	NN	O	O
an	NN	O	O
immunoglobulin	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
OFT-1	NN	O	O
appeared	NN	O	O
to	NN	O	O
have	NN	O	O
approximately	NN	O	O
the	NN	O	O
same	NN	O	O
transactivation	NN	O	O
ability	NN	O	O
as	NN	O	O
OTF-2	NN	O	B-protein
when	NN	O	O
normalized	NN	O	O
for	NN	O	O
binding	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
OTF-1	NN	O	B-protein
,	NN	O	O
without	NN	O	O
OTF-2	NN	O	B-protein
,	NN	O	O
is	NN	O	O
sufficient	NN	O	O
for	NN	O	O
transcription	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-DNA
genes	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
OTF-2	NN	O	B-protein
alone	NN	O	O
is	NN	O	O
not	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
B-cell-specific	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
.	NN	O	O

-DOCSTART-	O

Effects	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
on	NN	O	O
intralymphocytic	NN	O	O
sodium	NN	O	O
and	NN	O	O
potassium	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
vitro	NN	O	O
binding	NN	O	O
of	NN	O	O
aldosterone	NN	O	O
to	NN	O	O
mineralocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
on	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
and	NN	O	O
its	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
intracellular	NN	O	O
sodium	NN	O	O
and	NN	O	O
potassium	NN	O	O
concentrations	NN	O	O
of	NN	O	O
HML	NN	O	B-cell_type
have	NN	O	O
already	NN	O	O
been	NN	O	O
described	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
present	NN	O	O
paper	NN	O	O
this	NN	O	O
easily	NN	O	O
accessible	NN	O	O
human	NN	O	O
cell	NN	O	O
model	NN	O	O
was	NN	O	O
investigated	NN	O	O
in	NN	O	O
13	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
only	NN	O	O
four	NN	O	O
patients	NN	O	O
sodium	NN	O	O
in	NN	O	O
HML	NN	O	B-cell_type
without	NN	O	O
incubation	NN	O	O
was	NN	O	O
elevated	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
range	NN	O	O
for	NN	O	O
normal	NN	O	O
persons	NN	O	O
.	NN	O	O

A	NN	O	O
decrease	NN	O	O
of	NN	O	O
intracellular	NN	O	O
sodium	NN	O	O
or	NN	O	O
potassium	NN	O	O
occurred	NN	O	O
during	NN	O	O
incubation	NN	O	O
without	NN	O	O
aldosterone	NN	O	O
(	NN	O	O
P	NN	O	O
less	NN	O	O
than	NN	O	O
0.02	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
addition	NN	O	O
of	NN	O	O
1.4	NN	O	O
nM	NN	O	O
aldosterone	NN	O	O
did	NN	O	O
not	NN	O	O
prevent	NN	O	O
this	NN	O	O
loss	NN	O	O
of	NN	O	O
electrolytes	NN	O	O
as	NN	O	O
observed	NN	O	O
in	NN	O	O
normal	NN	O	O
persons	NN	O	O
.	NN	O	O

Plasma	NN	O	B-protein
renin	NN	O	I-protein
activity	NN	O	O
and	NN	O	O
aldosterone	NN	O	O
were	NN	O	O
not	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
electrolyte	NN	O	O
response	NN	O	O
and	NN	O	O
were	NN	O	O
within	NN	O	O
the	NN	O	O
normal	NN	O	O
limits	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
/cell	NN	O	O
were	NN	O	O
within	NN	O	O
or	NN	O	O
close	NN	O	O
to	NN	O	O
the	NN	O	O
normal	NN	O	O
range	NN	O	O
(	NN	O	O
n	NN	O	O
=	NN	O	O
9	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
independence	NN	O	O
of	NN	O	O
intracellular	NN	O	O
electrolytes	NN	O	O
from	NN	O	O
aldosterone	NN	O	O
despite	NN	O	O
a	NN	O	O
normal	NN	O	O
number	NN	O	O
of	NN	O	O
mineralocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
may	NN	O	O
reflect	NN	O	O
an	NN	O	O
impairment	NN	O	O
of	NN	O	O
the	NN	O	O
mineralocorticoid	NN	O	O
effector	NN	O	O
mechanism	NN	O	O
in	NN	O	O
the	NN	O	O
HML	NN	O	B-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
essential	NN	O	O
hypertension	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Differential	NN	O	O
diagnostic	NN	O	O
value	NN	O	O
of	NN	O	O
receptors	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
calcitriol	NN	O	O
)	NN	O	O
determination	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
children	NN	O	O
with	NN	O	O
rickets	NN	O	O
and	NN	O	O
rickets-like	NN	O	O
diseases	NN	O	O
]	NN	O	O

The	NN	O	O
authors	NN	O	O
provide	NN	O	O
the	NN	O	O
results	NN	O	O
of	NN	O	O
studying	NN	O	O
1	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
(	NN	O	O
calcitriol	NN	O	O
)	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
children	NN	O	O
with	NN	O	O
rickets	NN	O	O
and	NN	O	O
rickets-like	NN	O	O
diseases	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
proposed	NN	O	O
that	NN	O	O
the	NN	O	O
character	NN	O	O
of	NN	O	O
their	NN	O	O
expression	NN	O	O
under	NN	O	O
the	NN	O	O
influence	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D	NN	O	O
may	NN	O	O
be	NN	O	O
used	NN	O	O
with	NN	O	O
differential	NN	O	O
diagnostic	NN	O	O
purposes	NN	O	O
in	NN	O	O
view	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
with	NN	O	O
double-stranded	NN	O	O
phosphorothioate	NN	O	O
oligonucleotides	NN	O	O
.	NN	O	O

Alteration	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	NN	O	O
specific	NN	O	O
transcriptional	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
is	NN	O	O
necessary	NN	O	O
for	NN	O	O
determining	NN	O	O
how	NN	O	O
these	NN	O	O
factors	NN	O	O
participate	NN	O	O
in	NN	O	O
cellular	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
functions	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
can	NN	O	O
be	NN	O	O
antagonized	NN	O	O
by	NN	O	O
several	NN	O	O
methods	NN	O	O
,	NN	O	O
each	NN	O	O
with	NN	O	O
specific	NN	O	O
limitations	NN	O	O
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
sequence-specific	NN	O	B-protein
DNA-binding	NN	O	I-protein
proteins	NN	O	I-protein
was	NN	O	O
achieved	NN	O	O
with	NN	O	O
double-stranded	NN	O	O
(	NN	O	O
ds	NN	O	O
)	NN	O	O
phosphorothioate	NN	O	O
oligonucleotides	NN	O	O
that	NN	O	O
contained	NN	O	O
octamer	NN	O	B-DNA
or	NN	O	I-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
consensus	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
phosphorothioate	NN	O	O
oligonucleotides	NN	O	O
specifically	NN	O	O
bound	NN	O	O
either	NN	O	O
octamer	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
or	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
modified	NN	O	O
oligonucleotides	NN	O	O
accumulated	NN	O	O
in	NN	O	O
cells	NN	O	O
more	NN	O	O
effectively	NN	O	O
than	NN	O	O
standard	NN	O	O
ds	NN	O	O
oligonucleotides	NN	O	O
and	NN	O	O
modulated	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
a	NN	O	O
specific	NN	O	O
manner	NN	O	O
.	NN	O	O

Octamer-dependent	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	B-DNA
plasmid	NN	O	I-DNA
or	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	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
(	NN	O	I-DNA
HIV	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
was	NN	O	O
inhibited	NN	O	O
when	NN	O	O
the	NN	O	O
appropriate	NN	O	O
phosphorothioate	NN	O	O
oligonucleotide	NN	O	O
was	NN	O	O
added	NN	O	O
to	NN	O	O
a	NN	O	O
transiently	NN	O	B-cell_line
transfected	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

Addition	NN	O	O
of	NN	O	O
phosphorothioate	NN	O	O
oligonucleotides	NN	O	O
that	NN	O	O
contained	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
consensus	NN	O	I-DNA
to	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
leukemia	NN	O	I-cell_line
cells	NN	O	I-cell_line
inhibited	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
secretion	NN	O	O
to	NN	O	O
a	NN	O	O
degree	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
observed	NN	O	O
with	NN	O	O
a	NN	O	O
mutated	NN	O	B-DNA
octamer	NN	O	I-DNA
site	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

The	NN	O	O
ds	NN	O	O
phosphorothioate	NN	O	O
oligonucleotides	NN	O	O
probably	NN	O	O
compete	NN	O	O
for	NN	O	O
binding	NN	O	O
of	NN	O	O
specific	NN	O	O
transcription	NN	O	O
factors	NN	O	O
and	NN	O	O
may	NN	O	O
provide	NN	O	O
anti-viral	NN	O	O
,	NN	O	O
immunosuppressive	NN	O	O
,	NN	O	O
or	NN	O	O
other	NN	O	O
therapeutic	NN	O	O
effects	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
Effect	NN	O	O
of	NN	O	O
the	NN	O	O
regimen	NN	O	O
of	NN	O	O
kidney-tonifying	NN	O	O
and	NN	O	O
qi-invigorating	NN	O	O
on	NN	O	O
aging	NN	O	O
changes	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
]	NN	O	O

The	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentration	NN	O	O
and	NN	O	O
the	NN	O	O
sites	NN	O	O
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
(	NN	O	O
GCR	NN	O	B-protein
)	NN	O	O
in	NN	O	O
the	NN	O	O
peripheral	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
were	NN	O	O
measured	NN	O	O
in	NN	O	O
32	NN	O	O
healthy	NN	O	O
aged	NN	O	O
persons	NN	O	O
and	NN	O	O
13	NN	O	O
young	NN	O	O
adults	NN	O	O
.	NN	O	O

In	NN	O	O
animal	NN	O	O
experiment	NN	O	O
,	NN	O	O
GCR	NN	O	B-protein
of	NN	O	O
spleen	NN	O	B-cell_type
lymphocytic	NN	O	I-cell_type
cell	NN	O	I-cell_type
was	NN	O	O
also	NN	O	O
measured	NN	O	O
in	NN	O	O
18	NN	O	O
aged	NN	O	O
rats	NN	O	O
and	NN	O	O
9	NN	O	O
young	NN	O	O
rats	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
showed	NN	O	O
that	NN	O	O
GCR	NN	O	B-protein
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
the	NN	O	O
aged	NN	O	O
persons	NN	O	O
or	NN	O	O
rats	NN	O	O
than	NN	O	O
that	NN	O	O
in	NN	O	O
the	NN	O	O
young	NN	O	O
while	NN	O	O
the	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
level	NN	O	O
did	NN	O	O
n't	NN	O	O
change	NN	O	O
with	NN	O	O
aging	NN	O	O
.	NN	O	O

So	NN	O	O
we	NN	O	O
think	NN	O	O
that	NN	O	O
GCR	NN	O	B-protein
is	NN	O	O
more	NN	O	O
sensitive	NN	O	O
than	NN	O	O
the	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
level	NN	O	O
to	NN	O	O
reflect	NN	O	O
the	NN	O	O
aging	NN	O	O
change	NN	O	O
of	NN	O	O
the	NN	O	O
adrenal	NN	O	O
cortex	NN	O	O
function	NN	O	O
.	NN	O	O

After	NN	O	O
the	NN	O	O
treatment	NN	O	O
with	NN	O	O
the	NN	O	O
regimen	NN	O	O
of	NN	O	O
Kidney-tonifying	NN	O	O
and	NN	O	O
Qi-invigorating	NN	O	O
,	NN	O	O
the	NN	O	O
GCR	NN	O	B-protein
of	NN	O	O
the	NN	O	O
aged	NN	O	O
persons	NN	O	O
and	NN	O	O
rats	NN	O	O
was	NN	O	O
enhanced	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
this	NN	O	O
way	NN	O	O
,	NN	O	O
the	NN	O	O
function	NN	O	O
of	NN	O	O
the	NN	O	O
aged	NN	O	O
adrenal	NN	O	O
cortex	NN	O	O
was	NN	O	O
improved	NN	O	O
.	NN	O	O

-DOCSTART-	O

Interferon	NN	O	O
affects	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
cells	NN	O	O
of	NN	O	O
clinically	NN	O	O
sensitive	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
patients	NN	O	O
.	NN	O	O

Cytoplasmic	NN	O	O
protein	NN	O	O
extracts	NN	O	O
from	NN	O	O
chronic	NN	O	B-cell_type
myelogenous	NN	O	I-cell_type
leukemia	NN	O	I-cell_type
(	NN	O	I-cell_type
CML	NN	O	I-cell_type
)	NN	O	I-cell_type
cells	NN	O	I-cell_type
contained	NN	O	O
an	NN	O	O
activity	NN	O	O
that	NN	O	O
altered	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
of	NN	O	O
complexes	NN	O	O
formed	NN	O	O
between	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
enhancers	NN	O	O
of	NN	O	O
interferon	NN	O	O
(	NN	O	O
IFN	NN	O	O
)	NN	O	O
-inducible	NN	O	O
genes	NN	O	O
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
CML	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
IFN-alpha	NN	O	B-protein
diminished	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
CML	NN	O	B-protein
cytoplasmic	NN	O	I-protein
proteins	NN	O	I-protein
on	NN	O	O
these	NN	O	O
nuclear	NN	O	B-protein
protein-DNA	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
clinical	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
IFN-alpha	NN	O	B-protein
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
to	NN	O	O
the	NN	O	O
IFN	NN	O	B-protein
-induced	NN	O	O
change	NN	O	O
in	NN	O	O
the	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
protein-DNA	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

These	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
action	NN	O	O
of	NN	O	O
IFN-alpha	NN	O	B-protein
in	NN	O	O
CML	NN	O	O
may	NN	O	O
be	NN	O	O
linked	NN	O	O
to	NN	O	O
a	NN	O	O
pathway	NN	O	O
that	NN	O	O
can	NN	O	O
result	NN	O	O
in	NN	O	O
posttranslational	NN	O	O
modification	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
proteins	NN	O	I-protein
.	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
transactivates	NN	O	O
latent	NN	O	B-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
LMP1	NN	O	B-protein
.	NN	O	O

Several	NN	O	O
lines	NN	O	O
of	NN	O	O
evidence	NN	O	O
are	NN	O	O
compatible	NN	O	O
with	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	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
or	NN	O	O
leader	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
EBNA-LP	NN	O	B-protein
)	NN	O	O
affects	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
latent	NN	O	I-protein
infection	NN	O	I-protein
membrane	NN	O	I-protein
protein	NN	O	I-protein
LMP1	NN	O	B-protein
.	NN	O	O

We	NN	O	O
now	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
following	NN	O	O
.	NN	O	O

(	NN	O	O
i	NN	O	O
)	NN	O	O
Acute	NN	O	O
transfection	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
EBNA-2	NN	O	B-protein
under	NN	O	O
control	NN	O	O
of	NN	O	O
simian	NN	O	B-DNA
virus	NN	O	I-DNA
40	NN	O	I-DNA
or	NN	O	I-DNA
Moloney	NN	O	I-DNA
murine	NN	O	I-DNA
leukemia	NN	O	I-DNA
virus	NN	O	I-DNA
promoters	NN	O	I-DNA
resulted	NN	O	O
in	NN	O	O
increased	NN	O	O
LMP1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
P3HR-1-infected	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
and	NN	O	O
the	NN	O	O
P3HR-1	NN	O	B-cell_line
or	NN	O	O
Daudi	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

(	NN	O	O
ii	NN	O	O
)	NN	O	O
Transfection	NN	O	O
and	NN	O	O
expression	NN	O	O
of	NN	O	O
EBNA-LP	NN	O	B-protein
alone	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
LMP1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
did	NN	O	O
not	NN	O	O
act	NN	O	O
synergistically	NN	O	O
with	NN	O	O
EBNA-2	NN	O	B-protein
to	NN	O	O
affect	NN	O	O
LMP1	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

(	NN	O	O
iii	NN	O	O
)	NN	O	O
LMP1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
Daudi	NN	O	B-cell_line
and	NN	O	O
P3HR-1-infected	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
controlled	NN	O	O
at	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
level	NN	O	O
,	NN	O	O
and	NN	O	O
EBNA-2	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
Daudi	NN	O	B-cell_line
cells	NN	O	I-cell_line
increased	NN	O	O
LMP1	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

(	NN	O	O
iv	NN	O	O
)	NN	O	O
No	NN	O	O
other	NN	O	O
EBV	NN	O	B-DNA
genes	NN	O	I-DNA
were	NN	O	O
required	NN	O	O
for	NN	O	O
EBNA-2	NN	O	B-protein
transactivation	NN	O	O
of	NN	O	O
LMP1	NN	O	B-protein
since	NN	O	O
cotransfection	NN	O	O
of	NN	O	O
recombinant	NN	O	B-DNA
EBNA-2	NN	O	I-DNA
expression	NN	O	I-DNA
vectors	NN	O	I-DNA
and	NN	O	O
genomic	NN	O	O
LMP1	NN	O	B-DNA
DNA	NN	O	I-DNA
fragments	NN	O	I-DNA
enhanced	NN	O	O
LMP1	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
the	NN	O	O
EBV-negative	NN	O	B-cell_line
B-lymphoma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
BJAB	NN	O	B-cell_line
,	NN	O	O
Louckes	NN	O	B-cell_line
,	NN	O	O
and	NN	O	O
BL30	NN	O	B-cell_line
.	NN	O	O

(	NN	O	O
v	NN	O	O
)	NN	O	O
An	NN	O	O
EBNA-2-responsive	NN	O	B-DNA
element	NN	O	I-DNA
was	NN	O	O
found	NN	O	O
within	NN	O	O
the	NN	O	O
-512	NN	O	B-DNA
to	NN	O	I-DNA
+40	NN	O	I-DNA
LMP1	NN	O	I-DNA
DNA	NN	O	I-DNA
since	NN	O	O
this	NN	O	O
DNA	NN	O	O
linked	NN	O	O
to	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
was	NN	O	O
transactivated	NN	O	O
by	NN	O	O
cotransfection	NN	O	O
with	NN	O	O
an	NN	O	O
EBNA-2	NN	O	B-DNA
expression	NN	O	I-DNA
vector	NN	O	I-DNA
.	NN	O	O

(	NN	O	O
vi	NN	O	O
)	NN	O	O
The	NN	O	O
EBV	NN	O	O
type	NN	O	O
2	NN	O	O
EBNA-2	NN	O	B-protein
transactivated	NN	O	O
LMP1	NN	O	B-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
EBV	NN	O	B-protein
type	NN	O	I-protein
1	NN	O	I-protein
EBNA-2	NN	O	I-protein
.	NN	O	O

(	NN	O	O
vii	NN	O	O
)	NN	O	O
Two	NN	O	O
deletions	NN	O	O
within	NN	O	O
the	NN	O	O
EBNA-2	NN	O	B-protein
gene	NN	O	O
which	NN	O	O
rendered	NN	O	O
EBV	NN	O	O
transformation	NN	O	O
incompetent	NN	O	O
did	NN	O	O
not	NN	O	O
transactivate	NN	O	O
LMP1	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
a	NN	O	O
transformation-competent	NN	O	B-DNA
EBNA-2	NN	O	I-DNA
deletion	NN	O	I-DNA
mutant	NN	O	I-DNA
did	NN	O	O
transactivate	NN	O	O
LMP1	NN	O	B-protein
.	NN	O	O

LMP1	NN	O	B-protein
is	NN	O	O
a	NN	O	O
potent	NN	O	O
effector	NN	O	O
of	NN	O	O
B-lymphocyte	NN	O	O
activation	NN	O	O
and	NN	O	O
can	NN	O	O
act	NN	O	O
synergistically	NN	O	O
with	NN	O	O
EBNA-2	NN	O	B-protein
to	NN	O	O
induce	NN	O	O
cellular	NN	O	O
CD23	NN	O	O
gene	NN	O	O
expression	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
LMP1	NN	O	B-protein
amplifies	NN	O	O
the	NN	O	O
biological	NN	O	O
impact	NN	O	O
of	NN	O	O
EBNA-2	NN	O	B-protein
and	NN	O	O
underscores	NN	O	O
its	NN	O	O
central	NN	O	O
role	NN	O	O
in	NN	O	O
EBV-induced	NN	O	O
growth	NN	O	O
transformation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
,	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
c-myc	NN	O	B-DNA
genes	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
heat	NN	O	O
shock	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

The	NN	O	O
effect	NN	O	O
of	NN	O	O
heat	NN	O	O
shock	NN	O	O
on	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-DNA
protooncogenes	NN	O	I-DNA
c-fos	NN	O	B-DNA
,	NN	O	O
c-jun	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
c-myc	NN	O	B-DNA
was	NN	O	O
studied	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

Heat	NN	O	O
shock	NN	O	O
caused	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
c-fos	NN	O	O
and	NN	O	O
c-jun	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
and	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
in	NN	O	O
pre-B	NN	O	B-cell_line
(	NN	O	I-cell_line
Hyon	NN	O	I-cell_line
)	NN	O	I-cell_line
and	NN	O	I-cell_line
T	NN	O	I-cell_line
(	NN	O	I-cell_line
DND-41	NN	O	I-cell_line
)	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
freshly	NN	O	O
isolated	NN	O	O
normal	NN	O	O
human	NN	O	O
thymocytes	NN	O	O
.	NN	O	O

The	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
of	NN	O	O
these	NN	O	O
protooncogenes	NN	O	B-DNA
in	NN	O	O
Hyon	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
most	NN	O	O
pronounced	NN	O	O
at	NN	O	O
42	NN	O	O
and	NN	O	O
43	NN	O	O
degrees	NN	O	O
C	NN	O	O
;	NN	O	O
kinetic	NN	O	O
analysis	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
changes	NN	O	O
could	NN	O	O
be	NN	O	O
detected	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
of	NN	O	O
heat	NN	O	O
shock	NN	O	O
.	NN	O	O

Altered	NN	O	O
transcription	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
c-myc	NN	O	B-DNA
genes	NN	O	I-DNA
was	NN	O	O
the	NN	O	O
primary	NN	O	O
effect	NN	O	O
of	NN	O	O
heat	NN	O	O
shock	NN	O	O
.	NN	O	O

Secondarily	NN	O	O
,	NN	O	O
heat	NN	O	O
shock	NN	O	O
of	NN	O	O
Hyon	NN	O	B-cell_line
cells	NN	O	I-cell_line
stabilized	NN	O	O
the	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
level	NN	O	O
by	NN	O	O
increasing	NN	O	O
its	NN	O	O
half-life	NN	O	O
from	NN	O	O
24	NN	O	O
to	NN	O	O
45	NN	O	O
min	NN	O	O
.	NN	O	O

The	NN	O	O
overall	NN	O	O
effect	NN	O	O
of	NN	O	O
heat	NN	O	O
shock	NN	O	O
on	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
level	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
was	NN	O	O
a	NN	O	O
marked	NN	O	O
inhibition	NN	O	O
of	NN	O	O
its	NN	O	O
transcription	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
transcription	NN	O	O
of	NN	O	O
nuclear	NN	O	B-DNA
protooncogenes	NN	O	I-DNA
is	NN	O	O
regulated	NN	O	O
by	NN	O	O
heat	NN	O	O
shock	NN	O	O
indicating	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
nuclear	NN	O	B-DNA
protooncogenes	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
stress	NN	O	O
response	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Mapping	NN	O	O
of	NN	O	O
B-cell	NN	O	B-protein
epitopes	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
hepatitis	NN	O	I-protein
B	NN	O	I-protein
virus	NN	O	I-protein
X	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
immune	NN	O	O
response	NN	O	O
to	NN	O	O
the	NN	O	O
X	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
human	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
was	NN	O	O
studied	NN	O	O
by	NN	O	O
epitope	NN	O	O
mapping	NN	O	O
by	NN	O	O
using	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
MS2-HBx	NN	O	B-protein
fusion	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
.	NN	O	O

Antibodies	NN	O	O
in	NN	O	O
sera	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
and	NN	O	O
chronic	NN	O	O
HBV	NN	O	O
infection	NN	O	O
showed	NN	O	O
a	NN	O	O
multispecific	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Each	NN	O	O
serum	NN	O	O
contained	NN	O	O
antibodies	NN	O	B-protein
to	NN	O	O
a	NN	O	O
different	NN	O	O
set	NN	O	O
of	NN	O	O
epitopes	NN	O	B-protein
,	NN	O	O
which	NN	O	O
taken	NN	O	O
together	NN	O	O
cover	NN	O	O
most	NN	O	O
of	NN	O	O
the	NN	O	O
HBx	NN	O	B-protein
sequence	NN	O	I-protein
.	NN	O	O

Some	NN	O	O
of	NN	O	O
the	NN	O	O
epitopes	NN	O	O
were	NN	O	O
detectable	NN	O	O
only	NN	O	O
by	NN	O	O
immunoblotting	NN	O	O
with	NN	O	O
fusion	NN	O	B-protein
proteins	NN	O	I-protein
;	NN	O	O
others	NN	O	O
were	NN	O	O
detectable	NN	O	O
only	NN	O	O
by	NN	O	O
an	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
with	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
.	NN	O	O

The	NN	O	O
carboxy-terminal	NN	O	B-protein
half	NN	O	I-protein
of	NN	O	O
the	NN	O	O
HBx	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
preferentially	NN	O	O
recognized	NN	O	O
by	NN	O	O
antibodies	NN	O	B-protein
from	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
and	NN	O	O
contained	NN	O	O
a	NN	O	O
short	NN	O	O
immunodominant	NN	O	B-protein
antigenic	NN	O	I-protein
region	NN	O	I-protein
with	NN	O	O
at	NN	O	O
least	NN	O	O
two	NN	O	O
major	NN	O	O
nonoverlapping	NN	O	B-protein
epitopes	NN	O	I-protein
.	NN	O	O

Anti-	NN	O	O
HBx	NN	O	B-protein
antibody	NN	O	O
titers	NN	O	O
as	NN	O	O
revealed	NN	O	O
by	NN	O	O
peptide	NN	O	O
ELISAs	NN	O	O
were	NN	O	O
highest	NN	O	O
and	NN	O	O
most	NN	O	O
frequent	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
chronic	NN	O	O
hepatitis	NN	O	O
and	NN	O	O
usually	NN	O	O
low	NN	O	O
in	NN	O	O
acutely	NN	O	O
infected	NN	O	O
patients	NN	O	O
and	NN	O	O
asymptomatic	NN	O	O
carriers	NN	O	O
.	NN	O	O

The	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
remarkable	NN	O	O
qualitative	NN	O	O
and	NN	O	O
quantitative	NN	O	O
heterogeneity	NN	O	O
of	NN	O	O
the	NN	O	O
humoral	NN	O	O
HBx	NN	O	B-protein
immune	NN	O	O
response	NN	O	O
which	NN	O	O
can	NN	O	O
be	NN	O	O
monitored	NN	O	O
by	NN	O	O
HBx	NN	O	B-protein
-specific	NN	O	O
peptide	NN	O	O
ELISAs	NN	O	O
.	NN	O	O

Such	NN	O	O
tests	NN	O	O
may	NN	O	O
become	NN	O	O
useful	NN	O	O
diagnostic	NN	O	O
tools	NN	O	O
.	NN	O	O

-DOCSTART-	O

[	NN	O	O
The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
hydrocortisone	NN	O	O
on	NN	O	O
the	NN	O	O
chemotactic	NN	O	O
migration	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
]	NN	O	O

Random	NN	O	O
migration	NN	O	O
(	NN	O	O
RM	NN	O	O
)	NN	O	O
and	NN	O	O
chemotactic	NN	O	O
migration	NN	O	O
(	NN	O	O
ChtM	NN	O	O
)	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
to	NN	O	O
yeast	NN	O	O
activated	NN	O	O
serum	NN	O	O
were	NN	O	O
studied	NN	O	O
with	NN	O	O
the	NN	O	O
modified	NN	O	O
Boyden	NN	O	O
chamber	NN	O	O
method	NN	O	O
.	NN	O	O

Both	NN	O	O
RM	NN	O	O
and	NN	O	O
ChtM	NN	O	O
showed	NN	O	O
circadian	NN	O	O
rhythm	NN	O	O
.	NN	O	O

Leukocytes	NN	O	B-cell_type
migrated	NN	O	O
most	NN	O	O
rapidly	NN	O	O
at	NN	O	O
night	NN	O	O
.	NN	O	O

The	NN	O	O
difference	NN	O	O
between	NN	O	O
the	NN	O	O
peak	NN	O	O
(	NN	O	O
0	NN	O	O
:	NN	O	O
00	NN	O	O
)	NN	O	O
and	NN	O	O
trough	NN	O	O
values	NN	O	O
(	NN	O	O
8	NN	O	O
:	NN	O	O
00	NN	O	O
)	NN	O	O
of	NN	O	O
RM	NN	O	O
and	NN	O	O
ChtM	NN	O	O
was	NN	O	O
significant	NN	O	O
statistically	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

ChtM	NN	O	O
was	NN	O	O
inhibited	NN	O	O
by	NN	O	O
hydrocortisone	NN	O	O
(	NN	O	O
F	NN	O	O
)	NN	O	O
of	NN	O	O
physiological	NN	O	O
concentration	NN	O	O
(	NN	O	O
10	NN	O	O
(	NN	O	O
-9	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
which	NN	O	O
was	NN	O	O
dose-dependent	NN	O	O
and	NN	O	O
completely	NN	O	O
antagonized	NN	O	O
by	NN	O	O
the	NN	O	O
competitive	NN	O	O
antagonist	NN	O	O
,	NN	O	O
RU38486	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
was	NN	O	O
much	NN	O	O
more	NN	O	O
evident	NN	O	O
with	NN	O	O
higher	NN	O	O
dose	NN	O	O
(	NN	O	O
more	NN	O	O
than	NN	O	O
10	NN	O	O
(	NN	O	O
-5	NN	O	O
)	NN	O	O
mol/L	NN	O	O
)	NN	O	O
and	NN	O	O
it	NN	O	O
was	NN	O	O
also	NN	O	O
reversed	NN	O	O
by	NN	O	O
RU38486	NN	O	O
,	NN	O	O
but	NN	O	O
only	NN	O	O
partially	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
suggested	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
physiological	NN	O	O
regulator	NN	O	O
of	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
leukocytes	NN	O	B-cell_type
and	NN	O	O
its	NN	O	O
inhibitory	NN	O	O
action	NN	O	O
on	NN	O	O
ChtM	NN	O	O
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
antiinflammatory	NN	O	O
mechanisms	NN	O	O
of	NN	O	O
GC	NN	O	O
of	NN	O	O
pharmacological	NN	O	O
doses	NN	O	O
.	NN	O	O

The	NN	O	O
action	NN	O	O
of	NN	O	O
physiological	NN	O	O
and	NN	O	O
pharmacological	NN	O	O
concentration	NN	O	O
of	NN	O	O
GC	NN	O	O
may	NN	O	O
be	NN	O	O
mediated	NN	O	O
by	NN	O	O
low	NN	O	B-protein
affinity	NN	O	I-protein
specific	NN	O	I-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
of	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Heterogeneity	NN	O	O
of	NN	O	O
antigen	NN	O	B-protein
molecules	NN	O	I-protein
recognized	NN	O	O
by	NN	O	O
anti-tax1	NN	O	B-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
Lt-4	NN	O	B-protein
in	NN	O	O
cell	NN	O	O
lines	NN	O	O
bearing	NN	O	O
human	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
and	NN	O	O
related	NN	O	O
retroviruses	NN	O	O
.	NN	O	O

Using	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
,	NN	O	O
Lt-4	NN	O	B-protein
,	NN	O	O
directed	NN	O	O
against	NN	O	O
human	NN	O	B-protein
T	NN	O	I-protein
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
trans-activator	NN	O	I-protein
(	NN	O	I-protein
tax1	NN	O	I-protein
)	NN	O	I-protein
antigen	NN	O	I-protein
,	NN	O	O
we	NN	O	O
examined	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
tax1	NN	O	B-protein
and	NN	O	O
related	NN	O	O
antigens	NN	O	B-protein
in	NN	O	O
a	NN	O	O
variety	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
bearing	NN	O	O
HTLV-I	NN	O	O
and	NN	O	O
related	NN	O	O
retroviruses	NN	O	O
,	NN	O	O
simian	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
virus	NN	O	O
type	NN	O	O
I	NN	O	O
(	NN	O	O
STLV-I	NN	O	O
)	NN	O	O
and	NN	O	O
HTLV-II	NN	O	O
,	NN	O	O
by	NN	O	O
immunofluorescence	NN	O	O
and	NN	O	O
immunoblot	NN	O	O
assays	NN	O	O
.	NN	O	O

Lt-4	NN	O	B-protein
reacted	NN	O	O
with	NN	O	O
all	NN	O	O
HTLV-I-bearing	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
tested	NN	O	O
and	NN	O	O
five	NN	O	O
out	NN	O	O
of	NN	O	O
eight	NN	O	O
simian	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
bearing	NN	O	O
STLV-I	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
with	NN	O	O
an	NN	O	O
HTLV-II-bearing	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

Lt-4	NN	O	B-protein
detected	NN	O	O
40	NN	O	O
kd	NN	O	O
tax1	NN	O	B-protein
antigen	NN	O	I-protein
molecules	NN	O	I-protein
in	NN	O	O
most	NN	O	O
HTLV-I-bearing	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
except	NN	O	O
one	NN	O	O
cell	NN	O	O
line	NN	O	O
that	NN	O	O
expressed	NN	O	O
39	NN	O	B-protein
kd	NN	O	I-protein
tax1	NN	O	I-protein
antigen	NN	O	I-protein
.	NN	O	O

In	NN	O	O
the	NN	O	O
STLV-I-bearing	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
tax1-related	NN	O	B-protein
antigen	NN	O	I-protein
molecules	NN	O	I-protein
detected	NN	O	O
by	NN	O	O
Lt-4	NN	O	B-protein
were	NN	O	O
heterogeneous	NN	O	O
,	NN	O	O
having	NN	O	O
molecular	NN	O	O
weights	NN	O	O
in	NN	O	O
the	NN	O	O
range	NN	O	O
of	NN	O	O
36-41	NN	O	O
kd	NN	O	O
.	NN	O	O

-DOCSTART-	O

Characterization	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
enhancer-binding	NN	O	I-protein
proteins	NN	O	I-protein
from	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
Jurkat	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
transcription	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
is	NN	O	O
under	NN	O	O
the	NN	O	O
control	NN	O	O
of	NN	O	O
cellular	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
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

Among	NN	O	O
the	NN	O	O
protein-binding	NN	O	O
regions	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
is	NN	O	O
the	NN	O	O
transcription-enhancer	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
at	NN	O	O
least	NN	O	O
one	NN	O	O
inducible	NN	O	B-protein
,	NN	O	O
C1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
one	NN	O	O
constitutive	NN	O	B-protein
,	NN	O	I-protein
C2	NN	O	I-protein
,	NN	O	I-protein
protein	NN	O	I-protein
can	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
HIV	NN	O	B-DNA
enhancer	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
two	NN	O	O
proteins	NN	O	O
differ	NN	O	O
in	NN	O	O
their	NN	O	O
surface	NN	O	O
charge	NN	O	O
,	NN	O	O
since	NN	O	O
they	NN	O	O
are	NN	O	O
separable	NN	O	O
by	NN	O	O
anion-exchange	NN	O	O
chromatography	NN	O	O
.	NN	O	O

Bivalent	NN	O	O
cations	NN	O	O
such	NN	O	O
as	NN	O	O
Mg2+	NN	O	O
and	NN	O	O
Zn2+	NN	O	O
differentially	NN	O	O
affect	NN	O	O
their	NN	O	O
binding	NN	O	O
to	NN	O	O
oligonucleotides	NN	O	O
which	NN	O	O
contain	NN	O	O
the	NN	O	O
HIV-enhancer	NN	O	B-DNA
domain	NN	O	I-DNA
.	NN	O	O

Both	NN	O	O
C1	NN	O	B-protein
and	NN	O	O
C2	NN	O	B-protein
proteins	NN	O	I-protein
also	NN	O	O
bind	NN	O	O
to	NN	O	O
a	NN	O	O
similar	NN	O	O
sequence	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
interleukin-2-receptor	NN	O	B-DNA
alpha-subunit	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
inducible	NN	O	B-protein
C1	NN	O	I-protein
protein	NN	O	I-protein
was	NN	O	O
partially	NN	O	O
purified	NN	O	O
by	NN	O	O
three	NN	O	O
chromatographic	NN	O	O
steps	NN	O	O
and	NN	O	O
characterized	NN	O	O
by	NN	O	O
u.v.	NN	O	O
cross-linking	NN	O	O
as	NN	O	O
a	NN	O	O
47	NN	O	B-protein
kDa	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Differences	NN	O	O
in	NN	O	O
transcriptional	NN	O	B-DNA
enhancers	NN	O	I-DNA
of	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
HIV-2	NN	O	O
.	NN	O	O

Response	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
signals	NN	O	O
.	NN	O	O

T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
results	NN	O	O
in	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV	NN	O	O
replication	NN	O	O
and	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
one	NN	O	O
mechanism	NN	O	O
leading	NN	O	O
to	NN	O	O
the	NN	O	O
conversion	NN	O	O
from	NN	O	O
latent	NN	O	O
to	NN	O	O
active	NN	O	O
viral	NN	O	O
infection	NN	O	O
.	NN	O	O

In	NN	O	O
HIV-1	NN	O	O
,	NN	O	O
the	NN	O	O
sequences	NN	O	O
that	NN	O	O
respond	NN	O	O
to	NN	O	O
these	NN	O	O
signaling	NN	O	O
events	NN	O	O
are	NN	O	O
found	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
LTR	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
comprise	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
contains	NN	O	O
two	NN	O	O
conserved	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
for	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	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
)	NN	O	O
.	NN	O	O

The	NN	O	O
corresponding	NN	O	O
region	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
AIDS	NN	O	O
retrovirus	NN	O	O
,	NN	O	O
HIV-2	NN	O	O
,	NN	O	O
contains	NN	O	O
a	NN	O	O
conserved	NN	O	O
and	NN	O	O
a	NN	O	O
divergent	NN	O	O
NF	NN	O	B-DNA
kappa	NN	O	I-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	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
LTR	NN	O	I-DNA
responds	NN	O	O
better	NN	O	O
than	NN	O	O
the	NN	O	O
HIV-2	NN	O	B-DNA
LTR	NN	O	I-DNA
to	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
.	NN	O	O

These	NN	O	O
qualitative	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
are	NN	O	O
reproduced	NN	O	O
not	NN	O	O
only	NN	O	O
when	NN	O	O
HIV-1	NN	O	B-DNA
or	NN	O	I-DNA
HIV-2	NN	O	I-DNA
enhancers	NN	O	I-DNA
are	NN	O	O
placed	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
also	NN	O	O
when	NN	O	O
these	NN	O	O
enhancers	NN	O	B-DNA
are	NN	O	O
switched	NN	O	O
between	NN	O	O
their	NN	O	O
respective	NN	O	O
LTR	NN	O	B-DNA
.	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
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
binds	NN	O	O
to	NN	O	O
both	NN	O	O
conserved	NN	O	O
sites	NN	O	O
in	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
transcriptional	NN	O	I-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
only	NN	O	O
to	NN	O	O
the	NN	O	O
single	NN	O	O
conserved	NN	O	O
site	NN	O	O
in	NN	O	O
the	NN	O	O
HIV-2	NN	O	B-DNA
transcriptional	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Instead	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
activator	NN	O	B-protein
protein	NN	O	I-protein
3	NN	O	I-protein
binds	NN	O	O
to	NN	O	O
the	NN	O	O
divergent	NN	O	O
site	NN	O	O
in	NN	O	O
HIV-2	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
HIV-2	NN	O	O
are	NN	O	O
differentially	NN	O	O
regulated	NN	O	O
by	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
signals	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
difference	NN	O	O
may	NN	O	O
account	NN	O	O
for	NN	O	O
the	NN	O	O
longer	NN	O	O
period	NN	O	O
of	NN	O	O
viral	NN	O	O
latency	NN	O	O
observed	NN	O	O
with	NN	O	O
HIV-2	NN	O	O
than	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-X2	NN	O	B-protein
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
X2-box	NN	O	I-DNA
is	NN	O	O
activator	NN	O	B-protein
protein	NN	O	I-protein
1	NN	O	I-protein
.	NN	O	O

Expression	NN	O	O
cloning	NN	O	O
of	NN	O	O
c-Jun	NN	O	B-protein
.	NN	O	O

Human	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
MHC	NN	O	I-protein
Ag	NN	O	I-protein
are	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
glycoproteins	NN	O	I-protein
.	NN	O	O

Their	NN	O	O
constitutive	NN	O	O
expression	NN	O	O
is	NN	O	O
limited	NN	O	O
to	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
thymic	NN	O	B-cell_type
epithelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
many	NN	O	O
other	NN	O	O
cells	NN	O	O
their	NN	O	O
expression	NN	O	O
can	NN	O	O
be	NN	O	O
induced	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

Conserved	NN	O	O
upstream	NN	O	O
promoter	NN	O	O
sequences	NN	O	O
regulate	NN	O	O
this	NN	O	O
tissue-specific	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
.	NN	O	O

In	NN	O	O
the	NN	O	O
DRA	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
cis-acting	NN	O	B-DNA
regulatory	NN	O	I-DNA
motifs	NN	O	I-DNA
is	NN	O	O
the	NN	O	O
X2-box	NN	O	B-DNA
to	NN	O	O
which	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
X2	NN	O	I-protein
(	NN	O	O
NF-X2	NN	O	B-protein
)	NN	O	O
binds	NN	O	O
.	NN	O	O

Here	NN	O	O
,	NN	O	O
we	NN	O	O
present	NN	O	O
the	NN	O	O
isolation	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
full-length	NN	O	B-DNA
cDNA	NN	O	I-DNA
clone	NN	O	I-DNA
encoding	NN	O	O
NF-X2	NN	O	B-protein
.	NN	O	O

This	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
was	NN	O	O
isolated	NN	O	O
by	NN	O	O
expression	NN	O	O
cDNA	NN	O	B-DNA
cloning	NN	O	O
,	NN	O	O
and	NN	O	O
encodes	NN	O	O
the	NN	O	O
human	NN	O	B-protein
c-Jun	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
which	NN	O	O
together	NN	O	O
with	NN	O	O
c-Fos	NN	O	B-protein
forms	NN	O	O
the	NN	O	O
heterodimeric	NN	O	B-protein
activator	NN	O	I-protein
protein-1	NN	O	I-protein
transcription	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Whereas	NN	O	O
c-Fos/c-Jun	NN	O	B-protein
heterodimers	NN	O	I-protein
do	NN	O	O
not	NN	O	O
exist	NN	O	O
in	NN	O	O
B	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
they	NN	O	O
form	NN	O	O
and	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
X2-box	NN	O	B-DNA
in	NN	O	O
class	NN	O	B-cell_line
II	NN	O	I-cell_line
nonexpressing	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
c-Fos/c-Jun	NN	O	B-protein
heterodimers	NN	O	I-protein
might	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
repression	NN	O	O
of	NN	O	O
DRA	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Synthesis	NN	O	O
of	NN	O	O
4	NN	O	O
,	NN	O	O
19-disubstituted	NN	O	O
derivatives	NN	O	O
of	NN	O	O
DOC	NN	O	O
.	NN	O	O

Radioreceptor	NN	O	O
assay	NN	O	O
of	NN	O	O
some	NN	O	O
corticosteroid	NN	O	O
derivatives	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

Several	NN	O	O
new	NN	O	O
4	NN	O	O
,	NN	O	O
19-substituted	NN	O	O
steroids	NN	O	O
and	NN	O	O
previously	NN	O	O
synthesized	NN	O	O
corticosteroids	NN	O	O
were	NN	O	O
assayed	NN	O	O
for	NN	O	O
affinity	NN	O	O
to	NN	O	O
type	NN	O	B-protein
1	NN	O	I-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
.	NN	O	O

11	NN	O	O
beta	NN	O	O
,	NN	O	O
19-epoxy-4	NN	O	O
,	NN	O	O
21-dihydroxypregn-4-ene-3	NN	O	O
,	NN	O	O
20-dione	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
was	NN	O	O
hydrogenated	NN	O	O
with	NN	O	O
Pd-C	NN	O	O
to	NN	O	O
yield	NN	O	O
a	NN	O	O
mixture	NN	O	O
of	NN	O	O
all	NN	O	O
four	NN	O	O
dihydro	NN	O	O
derivatives	NN	O	O
5	NN	O	O
,	NN	O	O
accompanied	NN	O	O
by	NN	O	O
4	NN	O	O
,	NN	O	O
21-diacetoxy-11	NN	O	O
beta	NN	O	O
,	NN	O	O
19-epoxy-3-hydroxypregnan-20-one	NN	O	O
(	NN	O	O
6	NN	O	O
)	NN	O	O
and	NN	O	O
21-acetoxy-11	NN	O	O
beta	NN	O	O
,	NN	O	O
19-epoxy-4-hydroxypregnane-3	NN	O	O
,	NN	O	O
20-dione	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
.	NN	O	O

With	NN	O	O
hot	NN	O	O
acetic	NN	O	O
+	NN	O	O
p-toluenesulfonic	NN	O	O
acid	NN	O	O
5	NN	O	O
underwent	NN	O	O
rearrangement	NN	O	O
to	NN	O	O
21-acetoxy-11	NN	O	O
beta	NN	O	O
,	NN	O	O
19-epoxypregn-5-ene-4	NN	O	O
,	NN	O	O
20-dione	NN	O	O
(	NN	O	O
8	NN	O	O
)	NN	O	O
Pd-C	NN	O	O
hydrogenation	NN	O	O
of	NN	O	O
3	NN	O	O
,	NN	O	O
21-diacetoxy-5	NN	O	O
beta	NN	O	O
,	NN	O	O
19-cyclopregna-2	NN	O	O
,	NN	O	O
9	NN	O	O
(	NN	O	O
11	NN	O	O
)	NN	O	O
-diene-4	NN	O	O
,	NN	O	O
20-dione	NN	O	O
(	NN	O	O
10	NN	O	O
)	NN	O	O
gave	NN	O	O
3	NN	O	O
,	NN	O	O
21-diacetoxy-5	NN	O	O
beta	NN	O	O
,	NN	O	O
19-cyclopregn-5-ene-4	NN	O	O
,	NN	O	O
20-dione	NN	O	O
(	NN	O	O
11	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
9	NN	O	O
,	NN	O	O
11-dihydro	NN	O	O
derivative	NN	O	O
of	NN	O	O
the	NN	O	O
latter	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
10	NN	O	O
with	NN	O	O
warm	NN	O	O
HCl	NN	O	O
furnished	NN	O	O
19-chloro-4	NN	O	O
,	NN	O	O
21-dihydroxypregna-4	NN	O	O
,	NN	O	O
9	NN	O	O
(	NN	O	O
11	NN	O	O
)	NN	O	O
-diene-3	NN	O	O
,	NN	O	O
20-dione	NN	O	O
(	NN	O	O
13	NN	O	O
)	NN	O	O
.	NN	O	O

Pd-C	NN	O	O
hydrogenation	NN	O	O
of	NN	O	O
its	NN	O	O
diacetate	NN	O	O
14	NN	O	O
afforded	NN	O	O
the	NN	O	O
4	NN	O	O
,	NN	O	O
5-dihydro	NN	O	O
derivative	NN	O	O
18	NN	O	O
,	NN	O	O
19-chloro-21-acetoxypregn-9	NN	O	O
(	NN	O	O
11	NN	O	O
)	NN	O	O
-en-20-one	NN	O	O
(	NN	O	O
15	NN	O	O
)	NN	O	O
,	NN	O	O
its	NN	O	O
4-acetoxy	NN	O	O
derivative	NN	O	O
16	NN	O	O
and	NN	O	O
the	NN	O	O
3	NN	O	O
,	NN	O	O
4-diacetoxy	NN	O	O
derivative	NN	O	O
17	NN	O	O
.	NN	O	O

When	NN	O	O
tested	NN	O	O
in	NN	O	O
a	NN	O	O
radioreceptor	NN	O	O
assay	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
the	NN	O	O
synthesized	NN	O	O
compounds	NN	O	O
showed	NN	O	O
only	NN	O	O
low	NN	O	O
relative	NN	O	O
binding	NN	O	O
affinities	NN	O	O
(	NN	O	O
RBA	NN	O	O
)	NN	O	O
to	NN	O	O
type	NN	O	B-protein
1	NN	O	I-protein
receptor	NN	O	I-protein
,	NN	O	O
the	NN	O	O
highest	NN	O	O
being	NN	O	O
0.72	NN	O	O
%	NN	O	O
for	NN	O	O
13	NN	O	O
(	NN	O	O
aldosterone	NN	O	O
=	NN	O	O
100	NN	O	O
%	NN	O	O
)	NN	O	O
.	NN	O	O

For	NN	O	O
comparison	NN	O	O
,	NN	O	O
other	NN	O	O
RBA	NN	O	O
in	NN	O	O
this	NN	O	O
system	NN	O	O
were	NN	O	O
:	NN	O	O
19-noraldosterone	NN	O	O
,	NN	O	O
20	NN	O	O
%	NN	O	O
;	NN	O	O
18-deoxyaldosterone	NN	O	O
,	NN	O	O
5.8	NN	O	O
%	NN	O	O
;	NN	O	O
18-deoxy-19-noraldosterone	NN	O	O
,	NN	O	O
4.7	NN	O	O
%	NN	O	O
;	NN	O	O
18	NN	O	O
,	NN	O	O
21-anhydroaldosterone	NN	O	O
,	NN	O	O
0.37	NN	O	O
%	NN	O	O
;	NN	O	O
17-isoaldosterone	NN	O	O
,	NN	O	O
7.6	NN	O	O
%	NN	O	O
and	NN	O	O
apoaldosterone	NN	O	O
,	NN	O	O
4.3	NN	O	O
%	NN	O	O

-DOCSTART-	O

Cell	NN	O	O
type	NN	O	O
specificity	NN	O	O
and	NN	O	O
activation	NN	O	O
requirements	NN	O	O
for	NN	O	O
NFAT-1	NN	O	B-protein
(	NN	O	O
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
)	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
determined	NN	O	O
by	NN	O	O
a	NN	O	O
new	NN	O	O
method	NN	O	O
using	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
to	NN	O	O
assay	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
an	NN	O	O
individual	NN	O	O
nuclear	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	O
activated	NN	O	B-cell_type
T-cells	NN	O	I-cell_type
(	NN	O	O
NFAT-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
which	NN	O	O
is	NN	O	O
considered	NN	O	O
to	NN	O	O
be	NN	O	O
an	NN	O	O
important	NN	O	O
regulator	NN	O	O
in	NN	O	O
early	NN	O	O
T-cell	NN	O	O
activation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
developed	NN	O	O
a	NN	O	O
system	NN	O	O
to	NN	O	O
monitor	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
of	NN	O	O
NFAT-1	NN	O	B-protein
at	NN	O	O
the	NN	O	O
single	NN	O	O
cell	NN	O	O
level	NN	O	O
in	NN	O	O
whole	NN	O	O
animals	NN	O	O
.	NN	O	O

The	NN	O	O
system	NN	O	O
is	NN	O	O
based	NN	O	O
on	NN	O	O
the	NN	O	O
use	NN	O	O
of	NN	O	O
an	NN	O	O
oligomerized	NN	O	O
NFAT-1	NN	O	B-protein
binding	NN	O	O
motif	NN	O	O
that	NN	O	O
directs	NN	O	O
transcription	NN	O	B-protein
of	NN	O	I-protein
SV40	NN	O	I-protein
T-antigen	NN	O	I-protein
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

This	NN	O	O
report	NN	O	O
represents	NN	O	O
the	NN	O	O
first	NN	O	O
demonstration	NN	O	O
that	NN	O	O
a	NN	O	O
multimerized	NN	O	O
short	NN	O	O
binding	NN	O	O
motif	NN	O	O
can	NN	O	O
function	NN	O	O
appropriately	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

NFAT-1	NN	O	B-protein
activity	NN	O	O
had	NN	O	O
previously	NN	O	O
been	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
confined	NN	O	O
to	NN	O	O
activated	NN	O	B-cell_type
T-lymphocytes	NN	O	I-cell_type
upon	NN	O	O
release	NN	O	O
of	NN	O	O
intracellular	NN	O	B-cell_type
calcium	NN	O	I-cell_type
.	NN	O	O

By	NN	O	O
targeting	NN	O	O
NFAT-1	NN	O	B-protein
-dependent	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
we	NN	O	O
discovered	NN	O	O
new	NN	O	O
sites	NN	O	O
of	NN	O	O
NFAT-1	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

Besides	NN	O	O
in	NN	O	O
T-lymphocytes	NN	O	B-cell_type
NFAT-1	NN	O	B-protein
activity	NN	O	O
could	NN	O	O
also	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
T-lymphocyte-depleted	NN	O	B-cell_type
spleen	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
purified	NN	O	B-cell_type
B-lymphocytes	NN	O	I-cell_type
and	NN	O	O
requires	NN	O	O
agents	NN	O	O
that	NN	O	O
both	NN	O	O
release	NN	O	O
intracellular	NN	O	O
calcium	NN	O	O
and	NN	O	O
activate	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
.	NN	O	O

A	NN	O	O
difference	NN	O	O
in	NN	O	O
the	NN	O	O
time	NN	O	O
course	NN	O	O
of	NN	O	O
appearance	NN	O	O
of	NN	O	O
NFAT-1	NN	O	B-protein
activity	NN	O	O
between	NN	O	O
T-lymphocytes	NN	O	B-cell_type
and	NN	O	O
non-T-lymphocytes	NN	O	B-cell_type
was	NN	O	O
revealed	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
expression	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
a	NN	O	O
small	NN	O	O
population	NN	O	O
of	NN	O	O
cells	NN	O	O
in	NN	O	O
the	NN	O	O
dermis	NN	O	O
and	NN	O	O
some	NN	O	O
mice	NN	O	O
have	NN	O	O
developed	NN	O	O
skin	NN	O	O
lesions	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
the	NN	O	O
tissue	NN	O	O
pattern	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
NFAT-1	NN	O	B-protein
activity	NN	O	O
resembles	NN	O	O
the	NN	O	O
expression	NN	O	O
pattern	NN	O	O
described	NN	O	O
for	NN	O	O
HIV-LTR/tat	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
(	NN	O	O
Vogel	NN	O	O
,	NN	O	O
J.	NN	O	O
,	NN	O	O
Hinrichs	NN	O	O
,	NN	O	O
S.	NN	O	O
H.	NN	O	O
,	NN	O	O
Reynolds	NN	O	O
,	NN	O	O
R.	NN	O	O
K.	NN	O	O
,	NN	O	O
Luciw	NN	O	O
,	NN	O	O
P.	NN	O	O
A.	NN	O	O
,	NN	O	O
and	NN	O	O
Jay	NN	O	O
,	NN	O	O
G.	NN	O	O
(	NN	O	O
1988	NN	O	O
)	NN	O	O
Nature	NN	O	O
335	NN	O	O
,	NN	O	O
606-611	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
similarity	NN	O	O
in	NN	O	O
expression	NN	O	O
and	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
NFAT-1	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
functional	NN	O	B-DNA
sequences	NN	O	I-DNA
in	NN	O	O
HIV-LTR	NN	O	B-DNA
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
NFAT-1	NN	O	B-protein
in	NN	O	O
dermal	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-LTR	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
novel	NN	O	O
T-cell	NN	O	B-protein
protein	NN	O	I-protein
which	NN	O	O
recognizes	NN	O	O
a	NN	O	O
palindromic	NN	O	B-DNA
sequence	NN	O	I-DNA
in	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	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

Two	NN	O	O
major	NN	O	B-DNA
protein-binding	NN	O	I-DNA
sites	NN	O	I-DNA
within	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	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
have	NN	O	O
been	NN	O	O
identified	NN	O	O
.	NN	O	O

One	NN	O	O
(	NN	O	O
site	NN	O	B-DNA
B	NN	O	I-DNA
)	NN	O	O
contained	NN	O	O
a	NN	O	O
palindromic	NN	O	B-DNA
sequence	NN	O	I-DNA
with	NN	O	O
homology	NN	O	O
to	NN	O	O
steroid/thyroid	NN	O	B-DNA
hormone	NN	O	I-DNA
response	NN	O	I-DNA
elements	NN	O	I-DNA
but	NN	O	O
was	NN	O	O
distinct	NN	O	O
from	NN	O	O
previously	NN	O	O
described	NN	O	O
binding	NN	O	O
sites	NN	O	O
of	NN	O	O
this	NN	O	O
class	NN	O	O
.	NN	O	O

A	NN	O	O
novel	NN	O	O
T-cell	NN	O	B-protein
protein	NN	O	I-protein
recognized	NN	O	O
the	NN	O	O
palindromic	NN	O	B-DNA
sequence	NN	O	I-DNA
within	NN	O	O
site	NN	O	B-DNA
B	NN	O	I-DNA
and	NN	O	O
also	NN	O	O
bound	NN	O	O
estrogen-	NN	O	B-DNA
or	NN	O	I-DNA
thyroid	NN	O	I-DNA
hormone-response	NN	O	I-DNA
elements	NN	O	I-DNA
with	NN	O	O
lower	NN	O	O
affinity	NN	O	O
.	NN	O	O

A	NN	O	O
7-base-pair	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
site	NN	O	B-DNA
B	NN	O	I-DNA
palindrome	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
destroyed	NN	O	O
protein	NN	O	O
binding	NN	O	O
,	NN	O	O
resulted	NN	O	O
in	NN	O	O
increased	NN	O	O
expression	NN	O	O
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
1	NN	O	I-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
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Progesterone	NN	O	O
suppression	NN	O	O
of	NN	O	O
pregnancy	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
not	NN	O	O
mediated	NN	O	O
by	NN	O	O
glucocorticoid	NN	O	O
effect	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
investigated	NN	O	O
whether	NN	O	O
the	NN	O	O
suppressive	NN	O	O
effect	NN	O	O
of	NN	O	O
progesterone	NN	O	O
on	NN	O	O
pregnancy	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
specific	NN	O	O
progesterone	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
a	NN	O	O
competitive	NN	O	O
progesterone	NN	O	O
antagonist	NN	O	O
(	NN	O	O
RU486	NN	O	O
)	NN	O	O
and	NN	O	O
a	NN	O	O
specific	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
blocker	NN	O	O
(	NN	O	O
RU43044	NN	O	O
)	NN	O	O
were	NN	O	O
tested	NN	O	O
on	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
a	NN	O	O
blocking	NN	O	B-protein
factor	NN	O	I-protein
by	NN	O	O
progesterone-treated	NN	O	B-cell_line
pregnancy	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

RU	NN	O	O
486	NN	O	O
tested	NN	O	O
at	NN	O	O
an	NN	O	O
equal	NN	O	O
concentration	NN	O	O
as	NN	O	O
progesterone	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
the	NN	O	O
blocking	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
while	NN	O	O
RU	NN	O	O
43044	NN	O	O
was	NN	O	O
without	NN	O	O
effect	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
pregnancy	NN	O	O
,	NN	O	O
lymphocyte	NN	O	O
progesterone	NN	O	O
acts	NN	O	O
on	NN	O	O
specific	NN	O	O
progesterone	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
glucocorticoid	NN	O	B-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
are	NN	O	O
not	NN	O	O
involved	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
56-59-kilodalton	NN	O	B-protein
protein	NN	O	I-protein
identified	NN	O	O
in	NN	O	O
untransformed	NN	O	B-protein
steroid	NN	O	I-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
is	NN	O	O
a	NN	O	O
unique	NN	O	B-protein
protein	NN	O	I-protein
that	NN	O	O
exists	NN	O	O
in	NN	O	O
cytosol	NN	O	O
in	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
both	NN	O	O
the	NN	O	O
70-	NN	O	B-protein
and	NN	O	I-protein
90-kilodalton	NN	O	I-protein
heat	NN	O	I-protein
shock	NN	O	I-protein
proteins	NN	O	I-protein
.	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
9S	NN	O	B-protein
,	NN	O	O
untransformed	NN	O	B-protein
progestin	NN	O	I-protein
,	NN	O	I-protein
estrogen	NN	O	I-protein
,	NN	O	I-protein
androgen	NN	O	I-protein
,	NN	O	I-protein
and	NN	O	I-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
rabbit	NN	O	O
uterine	NN	O	O
and	NN	O	O
liver	NN	O	O
cytosols	NN	O	O
contain	NN	O	O
a	NN	O	O
59-kDa	NN	O	B-protein
protein	NN	O	I-protein
[	NN	O	O
Tai	NN	O	O
,	NN	O	O
P.K.	NN	O	O
,	NN	O	O
Maeda	NN	O	O
,	NN	O	O
Y.	NN	O	O
,	NN	O	O
Nakao	NN	O	O
,	NN	O	O
K.	NN	O	O
,	NN	O	O
Wakim	NN	O	O
,	NN	O	O
N.G.	NN	O	O
,	NN	O	O
Duhring	NN	O	O
,	NN	O	O
J.L.	NN	O	O
,	NN	O	O
&	NN	O	O
Faber	NN	O	O
,	NN	O	O
L.E.	NN	O	O
(	NN	O	O
1986	NN	O	O
)	NN	O	O
Biochemistry	NN	O	O
25	NN	O	O
,	NN	O	O
5269-5275	NN	O	O
]	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
work	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
KN	NN	O	I-protein
382/EC1	NN	O	I-protein
raised	NN	O	O
against	NN	O	O
the	NN	O	O
rabbit	NN	O	B-protein
59-kDa	NN	O	I-protein
protein	NN	O	I-protein
reacts	NN	O	O
with	NN	O	O
9S	NN	O	B-protein
,	NN	O	O
untransformed	NN	O	B-protein
glucocorticoid	NN	O	I-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
in	NN	O	O
cytosol	NN	O	O
prepared	NN	O	O
from	NN	O	O
human	NN	O	B-cell_line
IM-9	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
with	NN	O	O
4S	NN	O	B-protein
salt-transformed	NN	O	I-protein
receptors	NN	O	I-protein
.	NN	O	O

The	NN	O	O
human	NN	O	O
protein	NN	O	O
recognized	NN	O	O
by	NN	O	O
the	NN	O	O
EC1	NN	O	B-protein
antibody	NN	O	I-protein
is	NN	O	O
a	NN	O	O
56-kDa	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
p56	NN	O	B-protein
)	NN	O	O
of	NN	O	O
moderate	NN	O	O
abundance	NN	O	O
located	NN	O	O
predominantly	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
by	NN	O	O
indirect	NN	O	O
immunofluorescence	NN	O	O
.	NN	O	O

There	NN	O	O
are	NN	O	O
at	NN	O	O
least	NN	O	O
six	NN	O	O
isomorphs	NN	O	B-protein
of	NN	O	O
p56	NN	O	B-protein
by	NN	O	O
two-dimensional	NN	O	O
gel	NN	O	O
analysis	NN	O	O
.	NN	O	O

N-Terminal	NN	O	O
sequencing	NN	O	O
(	NN	O	O
20	NN	O	O
amino	NN	O	O
acids	NN	O	O
)	NN	O	O
shows	NN	O	O
that	NN	O	O
p56	NN	O	B-protein
is	NN	O	O
a	NN	O	O
unique	NN	O	B-protein
human	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

When	NN	O	O
p56	NN	O	B-protein
is	NN	O	O
immunoadsorbed	NN	O	O
from	NN	O	O
IM-9	NN	O	B-cell_line
cell	NN	O	I-cell_line
cytosol	NN	O	O
,	NN	O	O
both	NN	O	O
the	NN	O	O
70-	NN	O	B-protein
and	NN	O	I-protein
90-kDa	NN	O	I-protein
heat	NN	O	I-protein
shock	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
coadsorbed	NN	O	O
in	NN	O	O
an	NN	O	O
immune-specific	NN	O	O
manner	NN	O	O
.	NN	O	O

Neither	NN	O	O
heat	NN	O	B-protein
shock	NN	O	I-protein
protein	NN	O	I-protein
reacts	NN	O	O
directly	NN	O	O
with	NN	O	O
the	NN	O	O
EC1	NN	O	B-protein
antibody	NN	O	I-protein
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
p56	NN	O	B-protein
exists	NN	O	O
in	NN	O	O
cytosol	NN	O	O
in	NN	O	O
a	NN	O	O
higher	NN	O	O
order	NN	O	O
complex	NN	O	O
containing	NN	O	O
hsp70	NN	O	B-protein
and	NN	O	O
hsp90	NN	O	B-protein
,	NN	O	O
both	NN	O	O
of	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
have	NN	O	O
been	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
untransformed	NN	O	B-protein
steroid	NN	O	I-protein
receptors	NN	O	I-protein
.	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
proto-oncoprotein	NN	O	B-protein
ets-1	NN	O	B-protein
defines	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-DNA
activator	NN	O	I-DNA
sequence	NN	O	I-DNA
within	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
Moloney	NN	O	O
murine	NN	O	O
sarcoma	NN	O	O
virus	NN	O	O
.	NN	O	O

The	NN	O	O
ets	NN	O	B-DNA
proto-oncogene	NN	O	I-DNA
family	NN	O	I-DNA
is	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
sequence-related	NN	O	B-DNA
genes	NN	O	I-DNA
whose	NN	O	O
normal	NN	O	O
cellular	NN	O	O
function	NN	O	O
is	NN	O	O
unknown	NN	O	O
.	NN	O	O

In	NN	O	O
a	NN	O	O
study	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
proteins	NN	O	I-protein
involved	NN	O	O
in	NN	O	O
the	NN	O	O
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
murine	NN	O	O
retroviruses	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
have	NN	O	O
discovered	NN	O	O
that	NN	O	O
a	NN	O	O
member	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
encodes	NN	O	O
a	NN	O	O
sequence-specific	NN	O	B-protein
DNA-binding	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

A	NN	O	O
mouse	NN	O	B-DNA
ets-1	NN	O	I-DNA
cDNA	NN	O	I-DNA
clone	NN	O	I-DNA
was	NN	O	O
obtained	NN	O	O
by	NN	O	O
screening	NN	O	O
a	NN	O	O
mouse	NN	O	O
thymus	NN	O	O
cDNA	NN	O	O
expression	NN	O	O
library	NN	O	O
with	NN	O	O
a	NN	O	O
double-stranded	NN	O	B-DNA
oligonucleotide	NN	O	I-DNA
probe	NN	O	I-DNA
representing	NN	O	O
20	NN	O	O
bp	NN	O	O
of	NN	O	O
the	NN	O	O
Moloney	NN	O	O
murine	NN	O	O
sarcoma	NN	O	O
virus	NN	O	O
(	NN	O	O
MSV	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

The	NN	O	O
cDNA	NN	O	B-DNA
sequence	NN	O	I-DNA
has	NN	O	O
an	NN	O	O
813-bp	NN	O	B-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
whose	NN	O	O
predicted	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
is	NN	O	O
97.6	NN	O	O
%	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
272	NN	O	B-protein
carboxy-terminal	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
ets-1	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

The	NN	O	O
ORF	NN	O	B-DNA
was	NN	O	O
expressed	NN	O	O
in	NN	O	O
bacteria	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
30-kD	NN	O	B-protein
protein	NN	O	I-protein
product	NN	O	I-protein
was	NN	O	O
shown	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
by	NN	O	O
mobility-shift	NN	O	O
assays	NN	O	O
,	NN	O	O
Southwestern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
and	NN	O	O
methylation	NN	O	O
interference	NN	O	O
.	NN	O	O

A	NN	O	O
mutant	NN	O	B-DNA
LTR	NN	O	I-DNA
containing	NN	O	O
four	NN	O	O
base	NN	O	O
pair	NN	O	O
substitutions	NN	O	O
in	NN	O	O
the	NN	O	O
ets-1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
constructed	NN	O	O
and	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
have	NN	O	O
reduced	NN	O	O
binding	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Transcriptional	NN	O	O
efficiency	NN	O	O
of	NN	O	O
the	NN	O	O
MSV	NN	O	B-DNA
LTR	NN	O	I-DNA
promoter	NN	O	I-DNA
containing	NN	O	O
this	NN	O	O
disrupted	NN	O	O
ets-1	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
compared	NN	O	O
to	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
wild-type	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
mouse	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
culture	NN	O	O
,	NN	O	O
and	NN	O	O
15-	NN	O	O
to	NN	O	O
20-fold	NN	O	O
reduction	NN	O	O
in	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
was	NN	O	O
observed	NN	O	O
.	NN	O	O

We	NN	O	O
propose	NN	O	O
that	NN	O	O
ets-1	NN	O	B-protein
functions	NN	O	O
as	NN	O	O
a	NN	O	O
transcriptional	NN	O	O
activator	NN	O	O
of	NN	O	O
mammalian	NN	O	O
type-C	NN	O	O
retroviruses	NN	O	O
and	NN	O	O
speculate	NN	O	O
that	NN	O	O
ets-related	NN	O	B-DNA
genes	NN	O	I-DNA
constitute	NN	O	O
a	NN	O	O
new	NN	O	O
group	NN	O	O
of	NN	O	O
eukaryotic	NN	O	B-protein
DNA-binding	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Type	NN	O	B-protein
II	NN	O	I-protein
estrogen	NN	O	I-protein
binding	NN	O	I-protein
sites	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
:	NN	O	O
variations	NN	O	O
during	NN	O	O
the	NN	O	O
menstrual	NN	O	O
cycle	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
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
contain	NN	O	O
type	NN	O	O
II	NN	O	B-protein
estrogen	NN	O	I-protein
binding	NN	O	I-protein
sites	NN	O	I-protein
(	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
EBS	NN	O	I-protein
)	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
fluctuations	NN	O	O
of	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
EBS	NN	O	I-protein
during	NN	O	O
the	NN	O	O
menstrual	NN	O	O
cycle	NN	O	O
were	NN	O	O
analyzed	NN	O	O
in	NN	O	O
6	NN	O	O
normally	NN	O	O
menstruating	NN	O	O
women	NN	O	O
.	NN	O	O

Approximately	NN	O	O
3	NN	O	O
times	NN	O	O
higher	NN	O	O
levels	NN	O	O
of	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
EBS	NN	O	I-protein
were	NN	O	O
found	NN	O	O
in	NN	O	O
the	NN	O	O
periovulatory	NN	O	O
period	NN	O	O
with	NN	O	O
respect	NN	O	O
to	NN	O	O
both	NN	O	O
follicular	NN	O	O
and	NN	O	O
luteal	NN	O	O
phases	NN	O	O
.	NN	O	O

In	NN	O	O
postmenopausal	NN	O	O
women	NN	O	O
the	NN	O	O
mean	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
EBS	NN	O	I-protein
levels	NN	O	O
were	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
follicular	NN	O	O
phase	NN	O	O
of	NN	O	O
the	NN	O	O
cycle	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
in	NN	O	O
3	NN	O	O
postmenopausal	NN	O	O
patients	NN	O	O
a	NN	O	O
short	NN	O	O
course	NN	O	O
of	NN	O	O
estrogen	NN	O	O
or	NN	O	O
tamoxifen	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
marked	NN	O	O
increase	NN	O	O
of	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
EBS	NN	O	I-protein
levels	NN	O	O
.	NN	O	O

Tamoxifen	NN	O	O
was	NN	O	O
also	NN	O	O
found	NN	O	O
to	NN	O	O
compete	NN	O	O
with	NN	O	O
17	NN	O	O
beta-estradiol	NN	O	O
for	NN	O	O
type	NN	O	B-protein
II	NN	O	I-protein
EBS	NN	O	I-protein
in	NN	O	O
PBMC	NN	O	B-cell_type
,	NN	O	O
although	NN	O	O
to	NN	O	O
a	NN	O	O
lesser	NN	O	O
extent	NN	O	O
than	NN	O	O
diethylstilbestrol	NN	O	O
.	NN	O	O

-DOCSTART-	O

Mononuclear	NN	O	B-cell_type
leukocyte	NN	O	I-cell_type
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
binding	NN	O	O
characteristics	NN	O	O
and	NN	O	O
down-regulation	NN	O	O
in	NN	O	O
major	NN	O	O
depression	NN	O	O
.	NN	O	O

Some	NN	O	O
patients	NN	O	O
with	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
have	NN	O	O
elevated	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
and	NN	O	O
show	NN	O	O
failure	NN	O	O
to	NN	O	O
suppress	NN	O	O
cortisol	NN	O	O
secretion	NN	O	O
upon	NN	O	O
administration	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
DEX	NN	O	O
)	NN	O	O
,	NN	O	O
yet	NN	O	O
they	NN	O	O
do	NN	O	O
not	NN	O	O
have	NN	O	O
Cushingoid	NN	O	O
features	NN	O	O
.	NN	O	O

To	NN	O	O
study	NN	O	O
whether	NN	O	O
this	NN	O	O
represents	NN	O	O
glucocorticoid	NN	O	O
(	NN	O	O
GC	NN	O	O
)	NN	O	O
resistance	NN	O	O
,	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
-DEX-binding	NN	O	O
assays	NN	O	O
were	NN	O	O
used	NN	O	O
to	NN	O	O
measure	NN	O	O
,	NN	O	O
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
the	NN	O	O
GC	NN	O	B-protein
receptor	NN	O	I-protein
affinity	NN	O	O
(	NN	O	O
1/Kd	NN	O	O
)	NN	O	O
and	NN	O	O
number	NN	O	O
(	NN	O	O
Bmax	NN	O	O
)	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
of	NN	O	O
11	NN	O	O
MDD	NN	O	O
patients	NN	O	O
and	NN	O	O
15	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

No	NN	O	O
receptor	NN	O	O
abnormalities	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
MDD	NN	O	O
group	NN	O	O
;	NN	O	O
thus	NN	O	O
any	NN	O	O
cellular	NN	O	O
defect	NN	O	O
leading	NN	O	O
to	NN	O	O
a	NN	O	O
lack	NN	O	O
of	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
GC	NN	O	O
in	NN	O	O
the	NN	O	O
MDD	NN	O	O
patients	NN	O	O
,	NN	O	O
if	NN	O	O
present	NN	O	O
,	NN	O	O
probably	NN	O	O
lies	NN	O	O
beyond	NN	O	O
the	NN	O	O
initial	NN	O	O
receptor	NN	O	O
binding	NN	O	O
.	NN	O	O

DEX	NN	O	O
(	NN	O	O
1.0	NN	O	O
mg	NN	O	O
orally	NN	O	O
)	NN	O	O
was	NN	O	O
administered	NN	O	O
to	NN	O	O
study	NN	O	O
in	NN	O	O
vivo	NN	O	B-protein
GC	NN	O	I-protein
receptor	NN	O	I-protein
down-regulation	NN	O	O
.	NN	O	O

Compared	NN	O	O
to	NN	O	O
the	NN	O	O
control	NN	O	O
group	NN	O	O
,	NN	O	O
fewer	NN	O	O
depressed	NN	O	O
subjects	NN	O	O
down-regulated	NN	O	O
Bmax	NN	O	O
after	NN	O	O
DEX	NN	O	O
.	NN	O	O

By	NN	O	O
paired	NN	O	O
t-test	NN	O	O
,	NN	O	O
Bmax	NN	O	O
decreased	NN	O	O
significantly	NN	O	O
in	NN	O	O
the	NN	O	O
control	NN	O	O
group	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
depressed	NN	O	O
group	NN	O	O
.	NN	O	O

Receptor	NN	O	O
number	NN	O	O
on	NN	O	O
the	NN	O	O
control	NN	O	O
day	NN	O	O
did	NN	O	O
not	NN	O	O
correlate	NN	O	O
significantly	NN	O	O
with	NN	O	O
the	NN	O	O
degree	NN	O	O
of	NN	O	O
receptor	NN	O	O
down-regulation	NN	O	O
,	NN	O	O
severity	NN	O	O
of	NN	O	O
depression	NN	O	O
or	NN	O	O
cortisol	NN	O	O
concentrations	NN	O	O
across	NN	O	O
all	NN	O	O
the	NN	O	O
subjects	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
do	NN	O	O
not	NN	O	O
lend	NN	O	O
support	NN	O	O
to	NN	O	O
previous	NN	O	O
reports	NN	O	O
suggesting	NN	O	O
that	NN	O	O
GC	NN	O	O
resistance	NN	O	O
in	NN	O	O
MDD	NN	O	O
results	NN	O	O
from	NN	O	O
a	NN	O	O
GC	NN	O	O
receptor-binding	NN	O	O
abnormality	NN	O	O
,	NN	O	O
and	NN	O	O
they	NN	O	O
emphasize	NN	O	O
the	NN	O	O
importance	NN	O	O
of	NN	O	O
considering	NN	O	O
receptor	NN	O	O
studies	NN	O	O
in	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
GC-mediated	NN	O	O
cell	NN	O	O
processes	NN	O	O
in	NN	O	O
order	NN	O	O
to	NN	O	O
identify	NN	O	O
the	NN	O	O
exact	NN	O	O
cellular	NN	O	O
defect	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
leading	NN	O	O
to	NN	O	O
GC	NN	O	O
resistance	NN	O	O
.	NN	O	O

-DOCSTART-	O

Ras-related	NN	O	B-protein
GTP-binding	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
leukocyte	NN	O	B-cell_type
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

Many	NN	O	O
aspects	NN	O	O
of	NN	O	O
leukocyte	NN	O	B-cell_type
function	NN	O	O
are	NN	O	O
regulated	NN	O	O
by	NN	O	O
both	NN	O	O
heterotrimeric	NN	O	B-protein
and	NN	O	O
Ras-related	NN	O	B-protein
GTP-binding	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
but	NN	O	O
there	NN	O	O
is	NN	O	O
little	NN	O	O
definite	NN	O	O
information	NN	O	O
about	NN	O	O
their	NN	O	O
roles	NN	O	O
in	NN	O	O
the	NN	O	O
specialized	NN	O	O
processes	NN	O	O
utilized	NN	O	O
by	NN	O	O
leukocytes	NN	O	B-cell_type
for	NN	O	O
cell	NN	O	O
killing	NN	O	O
.	NN	O	O

Recent	NN	O	O
progress	NN	O	O
in	NN	O	O
understanding	NN	O	O
the	NN	O	O
regulation	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
by	NN	O	O
the	NN	O	O
Rac	NN	O	B-protein
GTP-binding	NN	O	I-protein
proteins	NN	O	I-protein
provides	NN	O	O
a	NN	O	O
basis	NN	O	O
for	NN	O	O
defining	NN	O	O
the	NN	O	O
operational	NN	O	O
characteristics	NN	O	O
of	NN	O	O
one	NN	O	O
such	NN	O	O
phagocyte	NN	O	O
system	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
clear	NN	O	O
from	NN	O	O
various	NN	O	O
studies	NN	O	O
that	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
NADPH	NN	O	B-protein
oxidase	NN	O	I-protein
can	NN	O	O
be	NN	O	O
modulated	NN	O	O
through	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
GTP-GDP	NN	O	O
state	NN	O	O
of	NN	O	O
Rac	NN	O	B-protein
.	NN	O	O

Proteins	NN	O	O
exist	NN	O	O
in	NN	O	O
leukocytes	NN	O	B-cell_type
able	NN	O	O
to	NN	O	O
modify	NN	O	O
GTP-binding	NN	O	B-protein
protein	NN	O	I-protein
function	NN	O	O
in	NN	O	O
this	NN	O	O
manner	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
activity	NN	O	O
may	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	NN	O	O
signals	NN	O	O
generated	NN	O	O
on	NN	O	O
phagocyte	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Proteins	NN	O	O
of	NN	O	O
the	NN	O	O
Ras	NN	O	B-protein
superfamily	NN	O	I-protein
are	NN	O	O
likely	NN	O	O
to	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
normal	NN	O	O
phagocyte	NN	O	O
functions	NN	O	O
through	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
modulate	NN	O	O
the	NN	O	O
assembly	NN	O	O
of	NN	O	O
actin	NN	O	B-protein
filaments	NN	O	I-protein
,	NN	O	O
direct	NN	O	O
vesicle	NN	O	O
trafficking	NN	O	O
and	NN	O	O
fusion	NN	O	O
,	NN	O	O
and	NN	O	O
so	NN	O	O
forth	NN	O	O
.	NN	O	O

-DOCSTART-	O

BSAP	NN	O	B-protein
:	NN	O	O
a	NN	O	O
key	NN	O	O
regulator	NN	O	O
of	NN	O	O
B-cell	NN	O	B-cell_type
development	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	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
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
Pax-gene	NN	O	B-DNA
family	NN	O	I-DNA
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
;	NN	O	O
in	NN	O	O
the	NN	O	O
lymphoid	NN	O	O
system	NN	O	O
,	NN	O	O
BSAP	NN	O	B-protein
is	NN	O	O
produced	NN	O	O
only	NN	O	O
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Here	NN	O	O
,	NN	O	O
Markus	NN	O	O
Neurath	NN	O	O
,	NN	O	O
Eckhard	NN	O	O
Stuber	NN	O	O
and	NN	O	O
Warren	NN	O	O
Strober	NN	O	O
describe	NN	O	O
the	NN	O	O
molecular	NN	O	O
structure	NN	O	O
of	NN	O	O
BSAP	NN	O	B-protein
and	NN	O	O
focus	NN	O	O
on	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
to	NN	O	O
regulate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
B-cell-specific	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

They	NN	O	O
propose	NN	O	O
that	NN	O	O
BSAP	NN	O	B-protein
is	NN	O	O
a	NN	O	O
key	NN	O	B-protein
protein	NN	O	I-protein
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
that	NN	O	O
it	NN	O	O
not	NN	O	O
only	NN	O	O
influence	NN	O	O
B-cell	NN	O	O
development	NN	O	O
but	NN	O	O
also	NN	O	O
influences	NN	O	O
the	NN	O	O
balance	NN	O	O
between	NN	O	O
B-cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
immunoglobulin	NN	O	B-protein
secretion	NN	O	O
at	NN	O	O
later	NN	O	O
stages	NN	O	O
of	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Down-regulation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
protein	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
activated	NN	O	O
human	NN	O	O
lymphocytes	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
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
Jan	NN	O	O
9	NN	O	O
;	NN	O	O
93	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
:	NN	O	O
524	NN	O	O
]	NN	O	O

The	NN	O	O
effect	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
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2	NN	O	O
)	NN	O	O
D3	NN	O	O
]	NN	O	O
,	NN	O	O
a	NN	O	O
steroid	NN	O	O
hormone	NN	O	O
with	NN	O	O
immunomodulating	NN	O	O
properties	NN	O	O
,	NN	O	O
on	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
proteins	NN	O	O
was	NN	O	O
examined	NN	O	O
in	NN	O	O
in	NN	O	B-cell_line
vitro	NN	O	I-cell_line
activated	NN	O	I-cell_line
normal	NN	O	I-cell_line
human	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
by	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
.	NN	O	O

Over	NN	O	O
a	NN	O	O
72-hr	NN	O	O
period	NN	O	O
of	NN	O	O
activation	NN	O	O
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
50-kDa	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
p50	NN	O	B-protein
,	NN	O	O
and	NN	O	O
its	NN	O	O
precursor	NN	O	O
,	NN	O	O
p105	NN	O	B-protein
,	NN	O	O
was	NN	O	O
increased	NN	O	O
progressively	NN	O	O
.	NN	O	O

When	NN	O	O
cells	NN	O	O
were	NN	O	O
activated	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
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
levels	NN	O	O
of	NN	O	O
the	NN	O	O
mature	NN	O	O
protein	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
its	NN	O	O
precursor	NN	O	O
were	NN	O	O
decreased	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
hormone	NN	O	O
on	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
was	NN	O	O
demonstrable	NN	O	O
in	NN	O	O
the	NN	O	O
cytosolic	NN	O	O
and	NN	O	O
nuclear	NN	O	O
compartments	NN	O	O
;	NN	O	O
it	NN	O	O
required	NN	O	O
between	NN	O	O
4	NN	O	O
and	NN	O	O
8	NN	O	O
hr	NN	O	O
and	NN	O	O
was	NN	O	O
specific	NN	O	O
,	NN	O	O
as	NN	O	O
25-hydroxyvitamin	NN	O	O
D3	NN	O	O
and	NN	O	O
24	NN	O	O
,	NN	O	O
25-dihydroxyvitamin	NN	O	O
D3	NN	O	O
were	NN	O	O
ineffective	NN	O	O
.	NN	O	O

Besides	NN	O	O
p50	NN	O	B-protein
,	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
decreased	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
another	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
protein	NN	O	I-protein
,	NN	O	O
namely	NN	O	O
c-rel	NN	O	B-protein
.	NN	O	O

In	NN	O	O
addition	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
decreased	NN	O	O
the	NN	O	O
abundance	NN	O	O
of	NN	O	O
a	NN	O	O
specific	NN	O	B-protein
DNA-protein	NN	O	I-protein
complex	NN	O	I-protein
formed	NN	O	O
upon	NN	O	O
incubation	NN	O	O
of	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
activated	NN	O	O
lymphocytes	NN	O	O
with	NN	O	O
a	NN	O	O
labeled	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA	NN	O	I-protein
binding	NN	O	I-protein
motif	NN	O	I-protein
.	NN	O	O

Further	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
inhibited	NN	O	O
the	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
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
transiently	NN	O	O
transfected	NN	O	O
with	NN	O	O
a	NN	O	O
construct	NN	O	B-DNA
containing	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
NF-kappa	NN	O	B-DNA
B	NN	O	I-DNA
binding	NN	O	I-DNA
sequence	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-DNA
kappa	NN	O	I-DNA
light	NN	O	I-DNA
chain	NN	O	I-DNA
gene	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
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
observations	NN	O	O
demonstrate	NN	O	O
directly	NN	O	O
that	NN	O	O
there	NN	O	O
is	NN	O	O
de	NN	O	O
novo	NN	O	O
synthesis	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
during	NN	O	O
human	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
this	NN	O	O
process	NN	O	O
is	NN	O	O
hormonally	NN	O	O
regulated	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
myeloid	NN	O	B-DNA
zinc	NN	O	I-DNA
finger	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
MZF-1	NN	O	B-DNA
,	NN	O	O
regulates	NN	O	O
the	NN	O	O
CD34	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

MZF-1	NN	O	B-DNA
is	NN	O	O
a	NN	O	O
C2H2	NN	O	B-DNA
zinc	NN	O	I-DNA
finger	NN	O	I-DNA
gene	NN	O	I-DNA
encoding	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
transcriptional	NN	O	I-protein
regulator	NN	O	I-protein
of	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
MZF-1	NN	O	B-DNA
protein	NN	O	O
contains	NN	O	O
13	NN	O	O
C2H2	NN	O	B-protein
zinc	NN	O	I-protein
fingers	NN	O	I-protein
arranged	NN	O	O
in	NN	O	O
bipartite	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domains	NN	O	I-protein
containing	NN	O	O
zinc	NN	O	B-protein
fingers	NN	O	I-protein
through	NN	O	O
4	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
carboxy-terminus	NN	O	B-protein
,	NN	O	O
5	NN	O	O
through	NN	O	O
13	NN	O	O
.	NN	O	O

We	NN	O	O
previously	NN	O	O
identified	NN	O	O
the	NN	O	O
DNA	NN	O	B-DNA
consensus	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
recognized	NN	O	O
by	NN	O	O
the	NN	O	O
two	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domains	NN	O	I-protein
.	NN	O	O

To	NN	O	O
assess	NN	O	O
the	NN	O	O
transcription	NN	O	O
regulatory	NN	O	O
function	NN	O	O
of	NN	O	O
MZF-1	NN	O	B-DNA
,	NN	O	O
the	NN	O	O
full-length	NN	O	O
MZF-1	NN	O	B-DNA
coding	NN	O	O
region	NN	O	O
was	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
the	NN	O	O
yeast	NN	O	B-protein
transactivator	NN	O	I-protein
GAL4	NN	O	I-protein
.	NN	O	O

The	NN	O	O
expression	NN	O	O
vector	NN	O	O
was	NN	O	O
cotransfected	NN	O	O
with	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
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
thymidine	NN	O	B-DNA
kinase	NN	O	I-DNA
promoter	NN	O	I-DNA
containing	NN	O	O
GAL4	NN	O	B-DNA
DNA	NN	O	I-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
into	NN	O	O
NIH	NN	O	B-cell_line
3T3	NN	O	I-cell_line
,	NN	O	O
293	NN	O	B-cell_line
,	NN	O	O
K562	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

MZF-1	NN	O	B-DNA
represses	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
via	NN	O	O
GAL4	NN	O	B-protein
binding	NN	O	O
sites	NN	O	O
in	NN	O	O
the	NN	O	O
nonhematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
NIH	NN	O	B-cell_line
3T3	NN	O	I-cell_line
and	NN	O	O
293	NN	O	B-cell_line
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
MZF-1	NN	O	B-DNA
activates	NN	O	O
CAT	NN	O	B-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
the	NN	O	O
hematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
K562	NN	O	B-cell_line
and	NN	O	O
Jurkat	NN	O	B-cell_line
.	NN	O	O

The	NN	O	O
MZF-1	NN	O	B-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
promoters	NN	O	B-DNA
of	NN	O	O
several	NN	O	O
genes	NN	O	B-DNA
expressed	NN	O	O
during	NN	O	O
myeloid	NN	O	O
differentiation	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
CD34	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

MZF-1	NN	O	B-DNA
transcriptional	NN	O	O
regulation	NN	O	O
of	NN	O	O
this	NN	O	O
physiologically	NN	O	B-DNA
relevant	NN	O	I-DNA
promoter	NN	O	I-DNA
was	NN	O	O
assessed	NN	O	O
in	NN	O	O
both	NN	O	O
hematopoietic	NN	O	B-cell_line
and	NN	O	O
nonhematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Recombinant	NN	O	B-protein
MZF-1	NN	O	I-protein
protein	NN	O	I-protein
specifically	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
CD34	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
.	NN	O	O

MZF-1	NN	O	B-DNA
expression	NN	O	O
vectors	NN	O	O
were	NN	O	O
cotransfected	NN	O	O
with	NN	O	O
the	NN	O	O
luciferase	NN	O	B-DNA
reporter	NN	O	I-DNA
plasmids	NN	O	I-DNA
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
CD34	NN	O	B-DNA
promoter	NN	O	I-DNA
into	NN	O	O
both	NN	O	O
nonhematopoietic	NN	O	B-cell_line
and	NN	O	I-cell_line
hematopoietic	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

As	NN	O	O
with	NN	O	O
the	NN	O	O
heterologous	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
domain	NN	O	I-protein
,	NN	O	O
MZF-1	NN	O	B-DNA
represses	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
in	NN	O	O
nonhematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
activates	NN	O	O
expression	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

Activation	NN	O	O
of	NN	O	O
CD34	NN	O	B-protein
expression	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
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
intact	NN	O	O
MZF-1	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
cell	NN	O	O
type-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
CD34	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
MZF-1	NN	O	B-DNA
suggests	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
tissue-specific	NN	O	O
regulators/adapters	NN	O	O
or	NN	O	O
differential	NN	O	O
MZF-1	NN	O	B-DNA
modifications	NN	O	O
that	NN	O	O
determine	NN	O	O
MZF-1	NN	O	B-DNA
transcriptional	NN	O	O
regulatory	NN	O	O
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
murine	NN	O	O
BCL6	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
induced	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
as	NN	O	O
an	NN	O	O
immediate	NN	O	B-DNA
early	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
chromosomal	NN	O	O
translocation	NN	O	O
involving	NN	O	O
3q27	NN	O	B-DNA
is	NN	O	O
often	NN	O	O
detected	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
B-cell	NN	O	I-cell_type
lymphomas	NN	O	I-cell_type
,	NN	O	O
especially	NN	O	O
diffuse	NN	O	O
lymphomas	NN	O	O
with	NN	O	O
a	NN	O	O
large-cell	NN	O	O
component	NN	O	O
.	NN	O	O

The	NN	O	O
BCL6	NN	O	B-DNA
gene	NN	O	I-DNA
has	NN	O	O
been	NN	O	O
isolated	NN	O	O
from	NN	O	O
the	NN	O	O
chromosomal	NN	O	O
breakpoint	NN	O	O
in	NN	O	O
these	NN	O	O
lymphomas	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
cloned	NN	O	O
the	NN	O	O
murine	NN	O	B-DNA
BCL6	NN	O	I-DNA
(	NN	O	I-DNA
mBCL6	NN	O	I-DNA
)	NN	O	I-DNA
cDNA	NN	O	I-DNA
from	NN	O	O
the	NN	O	O
muscle	NN	O	B-DNA
cDNA	NN	O	I-DNA
library	NN	O	I-DNA
using	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
BCL6	NN	O	I-DNA
(	NN	O	I-DNA
hBCL6	NN	O	I-DNA
)	NN	O	I-DNA
cDNA	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
probe	NN	O	O
.	NN	O	O

The	NN	O	O
predicted	NN	O	O
amino	NN	O	O
acid	NN	O	O
sequence	NN	O	O
was	NN	O	O
95	NN	O	O
%	NN	O	O
identical	NN	O	O
to	NN	O	O
that	NN	O	O
of	NN	O	O
hBCL6	NN	O	B-protein
.	NN	O	O

It	NN	O	O
contains	NN	O	O
six	NN	O	O
repeats	NN	O	O
of	NN	O	O
the	NN	O	O
Kruppel-like	NN	O	B-protein
zinc-finger	NN	O	I-protein
motif	NN	O	I-protein
that	NN	O	O
are	NN	O	O
completely	NN	O	O
identical	NN	O	O
to	NN	O	O
those	NN	O	O
of	NN	O	O
hBCL6	NN	O	B-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
BCL6	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
well	NN	O	O
conserved	NN	O	O
between	NN	O	O
humans	NN	O	O
and	NN	O	O
mice	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
mBCL6	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
ubiquitously	NN	O	O
detected	NN	O	O
in	NN	O	O
adult	NN	O	O
mouse	NN	O	O
tissues	NN	O	O
including	NN	O	O
lymphatic	NN	O	O
organs	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
it	NN	O	O
was	NN	O	O
induced	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
activated	NN	O	O
with	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
and	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
within	NN	O	O
30	NN	O	O
min	NN	O	O
after	NN	O	O
stimulation	NN	O	O
.	NN	O	O

This	NN	O	O
induction	NN	O	O
was	NN	O	O
not	NN	O	O
inhibited	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
a	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

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
BCL6	NN	O	B-protein
plays	NN	O	O
a	NN	O	O
role	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
as	NN	O	O
an	NN	O	O
immediate	NN	O	O
early	NN	O	O
gene	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	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
in	NN	O	O
B-cell	NN	O	O
development	NN	O	O
.	NN	O	O

The	NN	O	O
hierarchy	NN	O	O
of	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
in	NN	O	O
B-cell	NN	O	O
development	NN	O	O
has	NN	O	O
recently	NN	O	O
been	NN	O	O
analyzed	NN	O	O
by	NN	O	O
targeted	NN	O	O
gene	NN	O	O
inactivation	NN	O	O
in	NN	O	O
the	NN	O	O
mouse	NN	O	O
.	NN	O	O

In	NN	O	O
this	NN	O	O
manner	NN	O	O
,	NN	O	O
the	NN	O	O
paired	NN	O	B-DNA
box	NN	O	I-DNA
containing	NN	O	I-DNA
gene	NN	O	I-DNA
Pax-5	NN	O	B-DNA
,	NN	O	O
encoding	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
specific	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
BSAP	NN	O	B-protein
,	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
key	NN	O	O
role	NN	O	O
in	NN	O	O
early	NN	O	O
B	NN	O	O
lymphopoiesis	NN	O	O
.	NN	O	O

Other	NN	O	O
experimental	NN	O	O
strategies	NN	O	O
have	NN	O	O
implicated	NN	O	O
BSAP	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
,	NN	O	O
isotype	NN	O	O
switching	NN	O	O
and	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
immunoglobulin	NN	O	B-protein
heavy-chain	NN	O	I-protein
gene	NN	O	O
at	NN	O	O
late	NN	O	O
stages	NN	O	O
of	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
DNA-binding	NN	O	O
properties	NN	O	O
of	NN	O	O
two	NN	O	B-protein
heat	NN	O	I-protein
shock	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
HSF1	NN	O	B-protein
and	NN	O	O
HSF3	NN	O	B-protein
,	NN	O	O
are	NN	O	O
induced	NN	O	O
in	NN	O	O
the	NN	O	O
avian	NN	O	B-cell_line
erythroblast	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
HD6	NN	O	B-cell_line
.	NN	O	O

Avian	NN	O	O
cells	NN	O	O
express	NN	O	O
three	NN	O	O
heat	NN	O	B-DNA
shock	NN	O	I-DNA
transcription	NN	O	I-DNA
factor	NN	O	I-DNA
(	NN	O	I-DNA
HSF	NN	O	I-DNA
)	NN	O	I-DNA
genes	NN	O	I-DNA
corresponding	NN	O	O
to	NN	O	O
a	NN	O	O
novel	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	O
HSF3	NN	O	B-protein
,	NN	O	O
and	NN	O	O
homologs	NN	O	O
of	NN	O	O
mouse	NN	O	O
and	NN	O	O
human	NN	O	O
HSF1	NN	O	B-protein
and	NN	O	O
HSF2	NN	O	B-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
biochemical	NN	O	O
and	NN	O	O
cell	NN	O	O
biological	NN	O	O
properties	NN	O	O
of	NN	O	O
these	NN	O	O
HSFs	NN	O	B-protein
reveals	NN	O	O
that	NN	O	O
HSF3	NN	O	B-protein
has	NN	O	O
properties	NN	O	O
in	NN	O	O
common	NN	O	O
with	NN	O	O
both	NN	O	O
HSF1	NN	O	B-protein
and	NN	O	O
HSF2	NN	O	B-protein
and	NN	O	O
yet	NN	O	O
has	NN	O	O
features	NN	O	O
which	NN	O	O
are	NN	O	O
distinct	NN	O	O
from	NN	O	O
both	NN	O	O
.	NN	O	O

HSF3	NN	O	B-protein
is	NN	O	O
constitutively	NN	O	O
expressed	NN	O	O
in	NN	O	O
the	NN	O	O
erythroblast	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
HD6	NN	O	I-cell_line
,	NN	O	O
the	NN	O	O
lymphoblast	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
MSB	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
embryo	NN	O	B-cell_line
fibroblasts	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
yet	NN	O	O
its	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
is	NN	O	O
induced	NN	O	O
only	NN	O	O
upon	NN	O	O
exposure	NN	O	O
of	NN	O	O
HD6	NN	O	O
cells	NN	O	O
to	NN	O	O
heat	NN	O	O
shock	NN	O	O
.	NN	O	O

Acquisition	NN	O	O
of	NN	O	O
HSF3	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
HD6	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
oligomerization	NN	O	O
from	NN	O	O
a	NN	O	O
non-DNA-binding	NN	O	B-protein
dimer	NN	O	I-protein
to	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
trimer	NN	O	I-protein
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
effect	NN	O	O
of	NN	O	O
heat	NN	O	O
shock	NN	O	O
on	NN	O	O
HSF1	NN	O	B-protein
is	NN	O	O
oligomerization	NN	O	O
of	NN	O	O
an	NN	O	O
inert	NN	O	B-protein
monomer	NN	O	I-protein
to	NN	O	O
a	NN	O	O
DNA-binding	NN	O	B-protein
trimer	NN	O	I-protein
.	NN	O	O

Induction	NN	O	O
of	NN	O	O
HSF3	NN	O	B-protein
DNA-binding	NN	O	O
activity	NN	O	O
is	NN	O	O
delayed	NN	O	O
compared	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
HSF1	NN	O	B-protein
.	NN	O	O

As	NN	O	O
occurs	NN	O	O
for	NN	O	O
HSF1	NN	O	B-protein
,	NN	O	O
heat	NN	O	O
shock	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
translocation	NN	O	O
of	NN	O	O
HSF3	NN	O	B-protein
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

HSF	NN	O	O
exhibits	NN	O	O
the	NN	O	O
properties	NN	O	O
of	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
,	NN	O	O
as	NN	O	O
judged	NN	O	O
from	NN	O	O
the	NN	O	O
stimulatory	NN	O	O
activity	NN	O	O
of	NN	O	O
transiently	NN	O	B-protein
overexpressed	NN	O	I-protein
HSF3	NN	O	I-protein
measured	NN	O	O
by	NN	O	O
using	NN	O	O
a	NN	O	O
heat	NN	O	B-DNA
shock	NN	O	I-DNA
element-containing	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
and	NN	O	O
as	NN	O	O
independently	NN	O	O
assayed	NN	O	O
by	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
chimeric	NN	O	B-protein
GAL4-HSF3	NN	O	I-protein
protein	NN	O	I-protein
on	NN	O	O
a	NN	O	O
GAL4	NN	O	B-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
results	NN	O	O
reveal	NN	O	O
that	NN	O	O
HSF3	NN	O	B-protein
is	NN	O	O
negatively	NN	O	O
regulated	NN	O	O
in	NN	O	O
avian	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
acquires	NN	O	O
DNA-binding	NN	O	O
activity	NN	O	O
in	NN	O	O
certain	NN	O	O
cells	NN	O	O
upon	NN	O	O
heat	NN	O	O
shock	NN	O	O
.	NN	O	O

-DOCSTART-	O

Direct	NN	O	O
demonstration	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
dephosphorylation	NN	O	O
and	NN	O	O
nuclear	NN	O	O
localization	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_line
HT-2	NN	O	I-cell_line
cells	NN	O	I-cell_line
using	NN	O	O
a	NN	O	O
specific	NN	O	O
NFATp	NN	O	B-protein
polyclonal	NN	O	I-protein
antibody	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
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	B-protein
genes	NN	O	I-protein
,	NN	O	O
and	NN	O	O
NFAT	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
is	NN	O	O
stimulated	NN	O	O
following	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
.	NN	O	O

Several	NN	O	O
lines	NN	O	O
of	NN	O	O
evidence	NN	O	O
have	NN	O	O
suggested	NN	O	O
that	NN	O	O
NFAT	NN	O	B-protein
is	NN	O	O
a	NN	O	O
substrate	NN	O	O
for	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
serine/threonine	NN	O	B-protein
phosphatase	NN	O	I-protein
.	NN	O	O

Using	NN	O	O
a	NN	O	O
polyclonal	NN	O	O
antibody	NN	O	O
to	NN	O	O
murine	NN	O	B-protein
NFATp	NN	O	I-protein
,	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
of	NN	O	O
various	NN	O	O
mouse	NN	O	O
tissues	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
110-130-kDa	NN	O	O
NFATp	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
highly	NN	O	O
expressed	NN	O	O
in	NN	O	O
thymus	NN	O	O
and	NN	O	O
spleen	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
immunoprecipitated	NN	O	O
NFATp	NN	O	B-protein
from	NN	O	O
untreated	NN	O	O
HT-2	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
calcineurin	NN	O	B-protein
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
,	NN	O	O
demonstrating	NN	O	O
that	NN	O	O
NFATp	NN	O	B-protein
is	NN	O	O
an	NN	O	O
in	NN	O	O
vitro	NN	O	O
substrate	NN	O	O
for	NN	O	O
calcineurin	NN	O	B-protein
.	NN	O	O

NFATp	NN	O	B-protein
immunoprecipitated	NN	O	O
from	NN	O	O
32P-labeled	NN	O	B-cell_line
HT-2	NN	O	I-cell_line
cells	NN	O	I-cell_line
migrated	NN	O	O
as	NN	O	O
an	NN	O	O
approximately	NN	O	O
120-kDa	NN	O	O
protein	NN	O	O
that	NN	O	O
was	NN	O	O
localized	NN	O	O
to	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
.	NN	O	O

Treatment	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
ionomycin	NN	O	O
resulted	NN	O	O
in	NN	O	O
a	NN	O	O
decrease	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
and	NN	O	O
a	NN	O	O
loss	NN	O	O
of	NN	O	O
32P	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
NFATp	NN	O	B-protein
dephosphorylation	NN	O	O
.	NN	O	O

The	NN	O	O
dephosphorylation	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
was	NN	O	O
accompanied	NN	O	O
by	NN	O	O
localization	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
to	NN	O	O
the	NN	O	O
nuclear	NN	O	O
fraction	NN	O	O
.	NN	O	O

Both	NN	O	O
of	NN	O	O
these	NN	O	O
events	NN	O	O
were	NN	O	O
blocked	NN	O	O
by	NN	O	O
preincubation	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
FK506	NN	O	O
,	NN	O	O
a	NN	O	O
calcineurin	NN	O	B-protein
inhibitor	NN	O	O
,	NN	O	O
consistent	NN	O	O
with	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
NFATp	NN	O	B-protein
is	NN	O	O
a	NN	O	O
calcineurin	NN	O	B-protein
substrate	NN	O	O
in	NN	O	O
cells	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
and	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
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
NFATp	NN	O	B-protein
and	NN	O	O
NFATc	NN	O	B-protein
,	NN	O	O
in	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
:	NN	O	O
regulation	NN	O	O
upon	NN	O	O
CD16	NN	O	B-protein
ligand	NN	O	O
binding	NN	O	O
.	NN	O	O

The	NN	O	O
putative	NN	O	O
factors	NN	O	O
that	NN	O	O
couple	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
from	NN	O	O
surface	NN	O	O
receptors	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
synthesis	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
have	NN	O	O
not	NN	O	O
been	NN	O	O
elucidated	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
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
(	NN	O	O
NFATp	NN	O	B-protein
)	NN	O	O
,	NN	O	O
a	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
-sensitive	NN	O	I-protein
factor	NN	O	I-protein
that	NN	O	O
regulates	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
several	NN	O	O
cytokines	NN	O	B-protein
,	NN	O	O
mediates	NN	O	O
CD16	NN	O	B-protein
-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
genes	NN	O	I-protein
in	NN	O	O
human	NN	O	B-cell_type
NK	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CD16	NN	O	B-protein
(	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RIIIA	NN	O	I-protein
)	NN	O	O
-induced	NN	O	O
expression	NN	O	O
of	NN	O	O
cytokine	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
occurs	NN	O	O
via	NN	O	O
a	NN	O	O
CsA-sensitive	NN	O	O
and	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
CD16	NN	O	B-protein
ligands	NN	O	O
induces	NN	O	O
NFAT	NN	O	B-protein
-like	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
these	NN	O	O
cells	NN	O	O
,	NN	O	O
as	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
.	NN	O	O

This	NN	O	O
occurs	NN	O	O
with	NN	O	O
fast	NN	O	O
kinetics	NN	O	O
after	NN	O	O
stimulation	NN	O	O
,	NN	O	O
via	NN	O	O
a	NN	O	O
CsA-sensitive	NN	O	O
and	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
mechanism	NN	O	O
that	NN	O	O
does	NN	O	O
not	NN	O	O
require	NN	O	O
de	NN	O	O
novo	NN	O	O
protein	NN	O	O
synthesis	NN	O	O
.	NN	O	O

NK	NN	O	B-protein
cell	NN	O	I-protein
NFAT	NN	O	I-protein
is	NN	O	O
present	NN	O	O
in	NN	O	O
the	NN	O	O
cytosol	NN	O	O
of	NN	O	O
nonstimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
migrates	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
upon	NN	O	O
stimulation	NN	O	O
,	NN	O	O
and	NN	O	O
can	NN	O	O
associate	NN	O	O
with	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

Two	NN	O	O
distinct	NN	O	O
molecules	NN	O	O
,	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	O
NFATc	NN	O	B-protein
,	NN	O	O
have	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
mediate	NN	O	O
NFAT	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
of	NN	O	O
supershift	NN	O	O
assays	NN	O	O
using	NN	O	O
NFATp-	NN	O	B-protein
and	NN	O	I-protein
NFATc-	NN	O	I-protein
specific	NN	O	I-protein
antibodies	NN	O	I-protein
indicate	NN	O	O
that	NN	O	O
NK	NN	O	O
cell	NN	O	O
activation	NN	O	O
early	NN	O	O
after	NN	O	O
CD16	NN	O	B-protein
ligand	NN	O	O
binding	NN	O	O
involves	NN	O	O
primarily	NN	O	O
,	NN	O	O
if	NN	O	O
not	NN	O	O
exclusively	NN	O	O
,	NN	O	O
NFATp	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
shows	NN	O	O
that	NN	O	O
this	NN	O	O
has	NN	O	O
the	NN	O	O
same	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
(	NN	O	O
approximately	NN	O	O
120	NN	O	O
kD	NN	O	O
)	NN	O	O
as	NN	O	O
that	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
do	NN	O	O
not	NN	O	O
express	NN	O	O
NFATc	NN	O	B-protein
constitutively	NN	O	O
,	NN	O	O
but	NN	O	O
NFATc	NN	O	B-RNA
mRNA	NN	O	I-RNA
accumulation	NN	O	O
is	NN	O	O
induced	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
within	NN	O	O
2	NN	O	O
h	NN	O	O
of	NN	O	O
stimulation	NN	O	O
with	NN	O	O
CD16	NN	O	O
ligands	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
supershift	NN	O	O
assays	NN	O	O
using	NN	O	O
the	NN	O	O
available	NN	O	O
mAb	NN	O	B-protein
recognizing	NN	O	O
the	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
NFATc	NN	O	I-protein
revealed	NN	O	O
no	NN	O	O
detectable	NN	O	O
NFATc	NN	O	B-protein
protein	NN	O	O
in	NN	O	O
nuclear	NN	O	O
and	NN	O	O
cytoplasmic	NN	O	O
extracts	NN	O	O
from	NN	O	O
CD16-	NN	O	O
or	NN	O	O
phorbol	NN	O	B-cell_line
ester-stimulated	NN	O	I-cell_line
cells	NN	O	I-cell_line
at	NN	O	O
any	NN	O	O
time	NN	O	O
tested	NN	O	O
,	NN	O	O
up	NN	O	O
to	NN	O	O
4	NN	O	O
h	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
that	NN	O	O
both	NN	O	O
CsA-sensitive	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	O
NFATc	NN	O	B-protein
,	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
human	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
that	NN	O	O
their	NN	O	O
activation	NN	O	O
and/or	NN	O	O
expression	NN	O	O
can	NN	O	O
be	NN	O	O
regulated	NN	O	O
in	NN	O	O
primary	NN	O	O
cells	NN	O	O
by	NN	O	O
a	NN	O	O
single	NN	O	O
stimulus	NN	O	O
,	NN	O	O
that	NN	O	O
,	NN	O	O
in	NN	O	O
the	NN	O	O
case	NN	O	O
of	NN	O	O
CD16	NN	O	O
in	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
results	NN	O	O
in	NN	O	O
early	NN	O	O
activation	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
and	NN	O	O
subsequently	NN	O	O
induced	NN	O	O
expression	NN	O	O
of	NN	O	O
NFATc	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

-DOCSTART-	O

Interleukin	NN	O	O
2	NN	O	O
signaling	NN	O	O
involves	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
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
most	NN	O	O
important	NN	O	O
cytokines	NN	O	B-protein
involved	NN	O	O
in	NN	O	O
immune	NN	O	O
response	NN	O	O
regulation	NN	O	O
is	NN	O	O
interleukin	NN	O	O
2	NN	O	O
(	NN	O	O
IL-2	NN	O	O
)	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
proliferation	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
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

The	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
the	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
are	NN	O	O
propagated	NN	O	O
within	NN	O	O
cells	NN	O	O
are	NN	O	O
not	NN	O	O
understood	NN	O	O
.	NN	O	O

While	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
to	NN	O	O
its	NN	O	O
receptor	NN	O	O
was	NN	O	O
recently	NN	O	O
shown	NN	O	O
to	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
two	NN	O	O
kinases	NN	O	B-protein
,	NN	O	O
Jak-1	NN	O	B-protein
and	NN	O	O
Jak-3	NN	O	B-protein
,	NN	O	O
subsequent	NN	O	O
steps	NN	O	O
in	NN	O	O
the	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
that	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
specific	NN	O	O
genes	NN	O	O
had	NN	O	O
not	NN	O	O
been	NN	O	O
characterized	NN	O	O
.	NN	O	O

Since	NN	O	O
many	NN	O	O
cytokines	NN	O	B-protein
that	NN	O	O
activate	NN	O	O
Jak	NN	O	B-protein
kinases	NN	O	I-protein
also	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
activation	NN	O	O
of	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
the	NN	O	O
ability	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
to	NN	O	O
trigger	NN	O	O
Stat	NN	O	O
phosphorylation	NN	O	O
was	NN	O	O
examined	NN	O	O
.	NN	O	O

Exposure	NN	O	O
of	NN	O	O
activated	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
or	NN	O	O
of	NN	O	O
a	NN	O	O
natural	NN	O	B-cell_line
killer	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
(	NN	O	O
NKL	NN	O	B-cell_line
)	NN	O	O
to	NN	O	O
IL-2	NN	O	B-protein
leads	NN	O	O
to	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat1	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
Stat1	NN	O	O
beta	NN	O	O
,	NN	O	O
and	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
of	NN	O	O
two	NN	O	O
Stat-related	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
p94	NN	O	B-protein
and	NN	O	O
p95	NN	O	B-protein
.	NN	O	O

p94	NN	O	B-protein
and	NN	O	O
p95	NN	O	B-protein
share	NN	O	O
homology	NN	O	O
with	NN	O	O
Stat1	NN	O	B-protein
at	NN	O	O
the	NN	O	O
phosphorylation	NN	O	B-protein
site	NN	O	I-protein
and	NN	O	O
in	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
,	NN	O	O
but	NN	O	O
otherwise	NN	O	O
are	NN	O	O
immunologically	NN	O	O
distinct	NN	O	O
from	NN	O	O
Stat1	NN	O	B-protein
.	NN	O	O

These	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
found	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
to	NN	O	O
bind	NN	O	O
to	NN	O	O
a	NN	O	O
specific	NN	O	B-DNA
DNA	NN	O	I-DNA
sequence	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
a	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
IL-2	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
its	NN	O	O
receptor	NN	O	O
may	NN	O	O
activate	NN	O	O
specific	NN	O	B-DNA
genes	NN	O	I-DNA
involved	NN	O	O
in	NN	O	O
immune	NN	O	O
cell	NN	O	O
function	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
correlates	NN	O	O
with	NN	O	O
IFN-alpha	NN	O	B-protein
responsiveness	NN	O	O
in	NN	O	O
Philadelphia	NN	O	O
chromosome	NN	O	O
positive	NN	O	O
chronic	NN	O	O
myelogenous	NN	O	O
leukemia	NN	O	O
.	NN	O	O

This	NN	O	O
study	NN	O	O
evaluates	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
constitutive	NN	O	O
levels	NN	O	O
of	NN	O	O
oncogene	NN	O	B-DNA
and	NN	O	O
p53	NN	O	B-RNA
transcripts	NN	O	I-RNA
in	NN	O	O
chronic	NN	O	O
phase	NN	O	O
CML	NN	O	O
patients	NN	O	O
and	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
their	NN	O	O
modulations	NN	O	O
subsequent	NN	O	O
to	NN	O	O
in	NN	O	O
vivo	NN	O	O
therapy	NN	O	O
with	NN	O	O
rIFN-alpha	NN	O	B-protein
2c	NN	O	I-protein
.	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
bone	NN	O	B-cell_type
marrow	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
26	NN	O	O
patients	NN	O	O
were	NN	O	O
examined	NN	O	O
for	NN	O	O
c-fos	NN	O	O
,	NN	O	O
c-myc	NN	O	O
,	NN	O	O
p53	NN	O	O
and	NN	O	O
the	NN	O	O
hybrid	NN	O	O
bcr/abl	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

Results	NN	O	O
indicated	NN	O	O
that	NN	O	O
(	NN	O	O
i	NN	O	O
)	NN	O	O
constitutive	NN	O	O
c-fos	NN	O	B-RNA
transcript	NN	O	I-RNA
levels	NN	O	O
are	NN	O	O
significantly	NN	O	O
higher	NN	O	O
in	NN	O	O
patients	NN	O	O
subsequently	NN	O	O
responding	NN	O	O
to	NN	O	O
IFN-alpha	NN	O	B-protein
therapy	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
positively	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
lymphocytes	NN	O	B-cell_type
(	NN	O	O
r	NN	O	O
=	NN	O	O
0.6895	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
negatively	NN	O	O
with	NN	O	O
the	NN	O	O
proportion	NN	O	O
of	NN	O	O
immature	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
r	NN	O	O
=	NN	O	O
-0.568	NN	O	O
,	NN	O	O
p	NN	O	O
<	NN	O	O
0.01	NN	O	O
)	NN	O	O
contained	NN	O	O
in	NN	O	O
the	NN	O	O
pbmc	NN	O	B-cell_type
preparations	NN	O	O
tested	NN	O	O
,	NN	O	O
(	NN	O	O
ii	NN	O	O
)	NN	O	O
constitutive	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
of	NN	O	O
the	NN	O	O
hybrid	NN	O	B-DNA
bcr/abl	NN	O	I-DNA
,	NN	O	O
c-myc	NN	O	B-DNA
and	NN	O	O
p53	NN	O	B-DNA
are	NN	O	O
positively	NN	O	O
correlated	NN	O	O
with	NN	O	O
each	NN	O	O
other	NN	O	O
,	NN	O	O
but	NN	O	O
failed	NN	O	O
to	NN	O	O
relate	NN	O	O
to	NN	O	O
disease	NN	O	O
parameters	NN	O	O
,	NN	O	O
and	NN	O	O
(	NN	O	O
iii	NN	O	O
)	NN	O	O
acute	NN	O	O
and	NN	O	O
chronic	NN	O	O
in	NN	O	O
vivo	NN	O	O
exposure	NN	O	O
to	NN	O	O
IFN-alpha	NN	O	B-protein
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
upregulation	NN	O	O
of	NN	O	O
c-fos	NN	O	B-DNA
and	NN	O	O
downregulation	NN	O	O
of	NN	O	O
c-myc	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
in	NN	O	O
responder	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Menopause	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
a	NN	O	O
significant	NN	O	O
increase	NN	O	O
in	NN	O	O
blood	NN	O	O
monocyte	NN	O	O
number	NN	O	O
and	NN	O	O
a	NN	O	O
relative	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
estrogen	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
monocytes	NN	O	I-cell_type
.	NN	O	O

PROBLEM	NN	O	O
:	NN	O	O
The	NN	O	O
clinical	NN	O	O
significance	NN	O	O
of	NN	O	O
the	NN	O	O
differential	NN	O	O
expression	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
monocytes	NN	O	I-cell_type
was	NN	O	O
evaluated	NN	O	O
.	NN	O	O

METHOD	NN	O	O
:	NN	O	O
Two	NN	O	O
color	NN	O	O
flow	NN	O	O
cytometry	NN	O	O
analysis	NN	O	O
was	NN	O	O
used	NN	O	O
on	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
samples	NN	O	O
of	NN	O	O
young	NN	O	O
and	NN	O	O
postmenopausal	NN	O	O
females	NN	O	O
and	NN	O	O
postmenopausal	NN	O	O
females	NN	O	O
treated	NN	O	O
with	NN	O	O
estrogen	NN	O	O
replacement	NN	O	O
therapy	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
monocyte	NN	O	O
and	NN	O	O
lymphocyte	NN	O	O
counts	NN	O	O
and	NN	O	O
the	NN	O	O
blood	NN	O	O
estrogen	NN	O	O
levels	NN	O	O
of	NN	O	O
each	NN	O	O
patient	NN	O	O
were	NN	O	O
determine	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
During	NN	O	O
menopause	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
ER	NN	O	B-cell_type
positive	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
blood	NN	O	B-cell_type
monocyte	NN	O	I-cell_type
number	NN	O	O
,	NN	O	O
which	NN	O	O
declines	NN	O	O
following	NN	O	O
estrogen	NN	O	O
replacement	NN	O	O
therapy	NN	O	O
to	NN	O	O
values	NN	O	O
of	NN	O	O
the	NN	O	O
young	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
These	NN	O	O
findings	NN	O	O
suggest	NN	O	O
that	NN	O	O
estrogen	NN	O	O
modulates	NN	O	O
the	NN	O	O
monocyte	NN	O	B-cell_type
numbers	NN	O	O
and	NN	O	O
its	NN	O	O
effects	NN	O	O
may	NN	O	O
be	NN	O	O
mediated	NN	O	O
through	NN	O	O
the	NN	O	O
ER	NN	O	B-protein
in	NN	O	O
the	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Staphylococcal	NN	O	B-protein
enterotoxins	NN	O	I-protein
modulate	NN	O	O
interleukin	NN	O	B-protein
2	NN	O	I-protein
receptor	NN	O	I-protein
expression	NN	O	O
and	NN	O	O
ligand-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
Janus	NN	O	B-protein
protein-tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
Jak3	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
proteins	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Staphylococcal	NN	O	B-protein
enterotoxins	NN	O	I-protein
(	NN	O	O
SE	NN	O	B-protein
)	NN	O	O
stimulate	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
expressing	NN	O	O
the	NN	O	O
appropriate	NN	O	O
variable	NN	O	B-protein
region	NN	O	I-protein
beta	NN	O	I-protein
chain	NN	O	I-protein
of	NN	O	I-protein
(	NN	O	I-protein
V	NN	O	I-protein
beta	NN	O	I-protein
)	NN	O	I-protein
T-cell	NN	O	I-protein
receptors	NN	O	I-protein
and	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
several	NN	O	O
autoimmune	NN	O	O
diseases	NN	O	O
.	NN	O	O

Depending	NN	O	O
on	NN	O	O
costimulatory	NN	O	O
signals	NN	O	O
,	NN	O	O
SE	NN	O	B-protein
induce	NN	O	O
either	NN	O	O
proliferation	NN	O	O
or	NN	O	O
anergy	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
addition	NN	O	O
,	NN	O	O
SE	NN	O	B-protein
can	NN	O	O
induce	NN	O	O
an	NN	O	O
interleukin-2	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
)	NN	O	O
nonresponsive	NN	O	O
state	NN	O	O
and	NN	O	O
apoptosis	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
SE	NN	O	B-protein
induce	NN	O	O
dynamic	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
and	NN	O	O
signal	NN	O	O
transduction	NN	O	O
through	NN	O	O
the	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
beta	NN	O	I-protein
and	NN	O	I-protein
gamma	NN	O	I-protein
chains	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
in	NN	O	O
human	NN	O	B-cell_line
antigen-specific	NN	O	I-cell_line
CD4+	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
after	NN	O	O
4	NN	O	O
hr	NN	O	O
of	NN	O	O
exposure	NN	O	O
to	NN	O	O
SEA	NN	O	B-protein
and	NN	O	O
SEB	NN	O	B-protein
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
was	NN	O	O
down-regulated	NN	O	O
,	NN	O	O
IL-2R	NN	O	B-protein
gamma	NN	O	I-protein
was	NN	O	O
slightly	NN	O	O
up-regulated	NN	O	O
,	NN	O	O
while	NN	O	O
IL-2R	NN	O	B-protein
alpha	NN	O	I-protein
remained	NN	O	O
largely	NN	O	O
unaffected	NN	O	O
.	NN	O	O

The	NN	O	O
changes	NN	O	O
in	NN	O	O
the	NN	O	O
composition	NN	O	O
of	NN	O	O
IL-2Rs	NN	O	B-protein
were	NN	O	O
accompanied	NN	O	O
by	NN	O	O
inhibition	NN	O	O
of	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
the	NN	O	O
Janus	NN	O	B-protein
protein-tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
3	NN	O	I-protein
(	NN	O	O
Jak3	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
called	NN	O	O
Stat3	NN	O	B-protein
and	NN	O	O
Stat5	NN	O	B-protein
.	NN	O	O

In	NN	O	O
parallel	NN	O	O
experiments	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
-driven	NN	O	O
proliferation	NN	O	O
was	NN	O	O
inhibited	NN	O	O
significantly	NN	O	O
.	NN	O	O

After	NN	O	O
16	NN	O	O
hr	NN	O	O
of	NN	O	O
exposure	NN	O	O
to	NN	O	O
SE	NN	O	B-protein
,	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
remained	NN	O	O
low	NN	O	O
,	NN	O	O
while	NN	O	O
that	NN	O	O
of	NN	O	O
IL2R	NN	O	B-protein
alpha	NN	O	I-protein
and	NN	O	O
IL2R	NN	O	B-protein
gamma	NN	O	I-protein
was	NN	O	O
further	NN	O	O
up-regulated	NN	O	O
,	NN	O	O
and	NN	O	O
ligand-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak3	NN	O	B-protein
and	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
was	NN	O	O
partly	NN	O	O
normalized	NN	O	O
.	NN	O	O

Yet	NN	O	O
,	NN	O	O
IL-2	NN	O	B-protein
-driven	NN	O	O
proliferation	NN	O	O
remained	NN	O	O
profoundly	NN	O	O
inhibited	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
signaling	NN	O	O
events	NN	O	O
other	NN	O	O
than	NN	O	O
Jak3	NN	O	B-protein
/Stat	NN	O	B-protein
activation	NN	O	O
had	NN	O	O
also	NN	O	O
been	NN	O	O
changed	NN	O	O
following	NN	O	O
SE	NN	O	B-protein
stimulation	NN	O	O
.	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
that	NN	O	O
SE	NN	O	B-protein
can	NN	O	O
modulate	NN	O	O
IL-2R	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
signal	NN	O	O
transduction	NN	O	O
involving	NN	O	O
the	NN	O	O
Jak/Stat	NN	O	B-protein
pathway	NN	O	O
in	NN	O	O
CD4+	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
,	NN	O	O
enhanced	NN	O	O
granulopoiesis	NN	O	O
,	NN	O	O
and	NN	O	O
neonatal	NN	O	O
lethality	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
-deficient	NN	O	O
mice	NN	O	O
.	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
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
family	NN	O	I-protein
are	NN	O	O
controlled	NN	O	O
by	NN	O	O
inhibitory	NN	O	B-protein
I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
mainly	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

Apparently	NN	O	O
normal	NN	O	O
at	NN	O	O
birth	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
mice	NN	O	O
exhibit	NN	O	O
severe	NN	O	O
runting	NN	O	O
,	NN	O	O
skin	NN	O	O
defects	NN	O	O
,	NN	O	O
and	NN	O	O
extensive	NN	O	O
granulopoiesis	NN	O	O
postnatally	NN	O	O
,	NN	O	O
typically	NN	O	O
dying	NN	O	O
by	NN	O	O
8	NN	O	O
days	NN	O	O
.	NN	O	O

Hematopoietic	NN	O	O
tissues	NN	O	O
from	NN	O	O
these	NN	O	O
mice	NN	O	O
display	NN	O	O
elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
both	NN	O	O
nuclear	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
and	NN	O	O
mRNAs	NN	O	B-RNA
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
genes	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	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
elevation	NN	O	O
results	NN	O	O
in	NN	O	O
these	NN	O	O
phenotypic	NN	O	O
abnormalities	NN	O	O
because	NN	O	O
mice	NN	O	O
lacking	NN	O	O
both	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
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
show	NN	O	O
a	NN	O	O
dramatically	NN	O	O
delayed	NN	O	O
onset	NN	O	O
of	NN	O	O
abnormalities	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
hematopoietic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
I	NN	O	B-cell_line
kappa	NN	O	I-cell_line
B	NN	O	I-cell_line
alpha-/-	NN	O	I-cell_line
embryonic	NN	O	I-cell_line
fibroblasts	NN	O	I-cell_line
show	NN	O	O
minimal	NN	O	O
constitutive	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
normal	NN	O	O
signal-dependent	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
that	NN	O	O
is	NN	O	O
concomitant	NN	O	O
with	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
.	NN	O	O

Our	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
I	NN	O	O
kappa	NN	O	O
b	NN	O	O
beta	NN	O	O
,	NN	O	O
but	NN	O	O
not	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
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
signal-dependent	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
fibroblasts	NN	O	B-cell_type
.	NN	O	O

However	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
required	NN	O	O
for	NN	O	O
the	NN	O	O
postinduction	NN	O	O
repression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
fibroblasts	NN	O	B-cell_type
.	NN	O	O

These	NN	O	O
results	NN	O	O
define	NN	O	O
distinct	NN	O	O
roles	NN	O	O
for	NN	O	O
the	NN	O	O
two	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
and	NN	O	O
demonstrate	NN	O	O
the	NN	O	O
necessity	NN	O	O
for	NN	O	O
stringent	NN	O	O
control	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Interleukin-7	NN	O	B-protein
can	NN	O	O
induce	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
Jak	NN	O	B-protein
1	NN	O	I-protein
,	NN	O	O
Jak	NN	O	B-protein
3	NN	O	I-protein
and	NN	O	O
STAT	NN	O	B-protein
5	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
murine	NN	O	O
T	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
Janus	NN	O	B-protein
protein	NN	O	I-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
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
proteins	NN	O	I-protein
has	NN	O	O
recently	NN	O	O
been	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
signal	NN	O	O
transduction	NN	O	O
mechanism	NN	O	O
of	NN	O	O
several	NN	O	O
cytokines	NN	O	B-protein
.	NN	O	O

IL-7	NN	O	B-protein
was	NN	O	O
observed	NN	O	O
to	NN	O	O
induce	NN	O	O
a	NN	O	O
rapid	NN	O	O
and	NN	O	O
dose-dependent	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak	NN	O	B-protein
1	NN	O	I-protein
and	NN	O	O
Jak	NN	O	B-protein
3	NN	O	I-protein
and	NN	O	O
concomitantly	NN	O	O
,	NN	O	O
the	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
of	NN	O	O
multiple	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
utilized	NN	O	O
by	NN	O	O
IL-7	NN	O	B-protein
were	NN	O	O
identical	NN	O	O
to	NN	O	O
those	NN	O	O
induced	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
could	NN	O	O
be	NN	O	O
identified	NN	O	O
as	NN	O	O
various	NN	O	O
STAT	NN	O	B-protein
5	NN	O	I-protein
isoforms	NN	O	I-protein
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
both	NN	O	O
Jak	NN	O	B-protein
1	NN	O	I-protein
and	NN	O	I-protein
3	NN	O	I-protein
,	NN	O	O
and	NN	O	O
STAT	NN	O	B-protein
5	NN	O	I-protein
activity	NN	O	O
strongly	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
growth-promoting	NN	O	O
effects	NN	O	O
of	NN	O	O
IL-7	NN	O	B-protein
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
signal	NN	O	O
transduction	NN	O	O
mechanism	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
IL-7	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cytokine	NN	O	B-protein
-modulating	NN	O	O
activity	NN	O	O
of	NN	O	O
tepoxalin	NN	O	O
,	NN	O	O
a	NN	O	O
new	NN	O	O
potential	NN	O	O
antirheumatic	NN	O	O
.	NN	O	O

Tepoxalin	NN	O	O
is	NN	O	O
a	NN	O	O
new	NN	O	O
dual	NN	O	O
cyclooxygenase/5-lipoxygenase	NN	O	O
anti-inflammatory	NN	O	O
compound	NN	O	O
currently	NN	O	O
under	NN	O	O
clinical	NN	O	O
investigation	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
possess	NN	O	O
anti-inflammatory	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
animal	NN	O	O
models	NN	O	O
and	NN	O	O
more	NN	O	O
recently	NN	O	O
to	NN	O	O
inhibit	NN	O	O
IL-2	NN	O	B-protein
induced	NN	O	O
signal	NN	O	O
transduction	NN	O	O
.	NN	O	O

The	NN	O	O
current	NN	O	O
study	NN	O	O
was	NN	O	O
conducted	NN	O	O
to	NN	O	O
evaluate	NN	O	O
the	NN	O	O
cytokine	NN	O	B-protein
modulating	NN	O	O
activity	NN	O	O
of	NN	O	O
tepoxalin	NN	O	O
and	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
iron	NN	O	O
in	NN	O	O
these	NN	O	O
effects	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
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
with	NN	O	O
OKT3/PMA	NN	O	O
,	NN	O	O
tepoxalin	NN	O	O
inhibited	NN	O	O
lymphocyte	NN	O	B-cell_type
proliferation	NN	O	O
with	NN	O	O
an	NN	O	O
IC50	NN	O	O
of	NN	O	O
6	NN	O	O
microM	NN	O	O
.	NN	O	O

Additionally	NN	O	O
,	NN	O	O
it	NN	O	O
inhibited	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
LTB4	NN	O	B-protein
(	NN	O	O
IC50	NN	O	O
=	NN	O	O
0.5	NN	O	O
microM	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
cytokines	NN	O	B-protein
IL-2	NN	O	B-protein
,	NN	O	O
IL-6	NN	O	B-protein
and	NN	O	O
TNF	NN	O	B-protein
alpha	NN	O	I-protein
(	NN	O	O
IC50	NN	O	O
=	NN	O	O
10-12	NN	O	O
microM	NN	O	O
)	NN	O	O
.	NN	O	O

Cytotoxicity	NN	O	O
was	NN	O	O
not	NN	O	O
demonstrated	NN	O	O
at	NN	O	O
these	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Add-back	NN	O	O
experiments	NN	O	O
with	NN	O	O
either	NN	O	O
cytokines	NN	O	B-protein
(	NN	O	O
IL-2	NN	O	B-protein
or	NN	O	O
IL-6	NN	O	B-protein
)	NN	O	O
,	NN	O	O
LTB4	NN	O	B-protein
or	NN	O	O
conditioned	NN	O	O
media	NN	O	O
failed	NN	O	O
to	NN	O	O
restore	NN	O	O
the	NN	O	O
proliferative	NN	O	O
response	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
tepoxalin	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
the	NN	O	O
concurrent	NN	O	O
addition	NN	O	O
of	NN	O	O
iron	NN	O	O
(	NN	O	O
in	NN	O	O
the	NN	O	O
form	NN	O	O
of	NN	O	O
ferrous	NN	O	O
or	NN	O	O
ferric	NN	O	O
chloride	NN	O	O
and	NN	O	O
other	NN	O	O
iron	NN	O	O
salts	NN	O	O
)	NN	O	O
reversed	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
proliferation	NN	O	O
caused	NN	O	O
by	NN	O	O
tepoxalin	NN	O	O
.	NN	O	O

Tepoxalin	NN	O	O
also	NN	O	O
inhibits	NN	O	O
the	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
transcription	NN	O	B-protein
factor	NN	O	I-protein
which	NN	O	O
acts	NN	O	O
on	NN	O	O
several	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

Tepoxalin	NN	O	O
's	NN	O	O
effect	NN	O	O
on	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
is	NN	O	O
also	NN	O	O
reversed	NN	O	O
by	NN	O	O
the	NN	O	O
addition	NN	O	O
of	NN	O	O
iron	NN	O	O
salts	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
action	NN	O	O
of	NN	O	O
tepoxalin	NN	O	O
to	NN	O	O
inhibit	NN	O	O
proliferation	NN	O	O
in	NN	O	O
PBMC	NN	O	B-cell_type
may	NN	O	O
be	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
due	NN	O	O
to	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
reduce	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
available	NN	O	O
iron	NN	O	O
resulting	NN	O	O
in	NN	O	O
decreased	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
and	NN	O	O
subsequent	NN	O	O
inhibition	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
.	NN	O	O

-DOCSTART-	O

N-	NN	O	B-protein
and	NN	O	I-protein
C-terminal	NN	O	I-protein
sequences	NN	O	I-protein
control	NN	O	O
degradation	NN	O	O
of	NN	O	O
MAD3/I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
inducers	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

The	NN	O	O
proteolytic	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
inhibitory	NN	O	B-protein
protein	NN	O	I-protein
MAD3/I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
in	NN	O	O
response	NN	O	O
to	NN	O	O
extracellular	NN	O	O
stimulation	NN	O	O
is	NN	O	O
a	NN	O	O
prerequisite	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
transcription	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
human	NN	O	B-protein
I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
stable	NN	O	O
transfectants	NN	O	O
of	NN	O	O
mouse	NN	O	B-cell_line
70Z/3	NN	O	I-cell_line
cells	NN	O	I-cell_line
shows	NN	O	O
that	NN	O	O
,	NN	O	O
as	NN	O	O
for	NN	O	O
the	NN	O	O
endogenous	NN	O	O
murine	NN	O	O
protein	NN	O	O
,	NN	O	O
exogenous	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
degraded	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
inducers	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
such	NN	O	O
as	NN	O	O
phorbol	NN	O	O
myristate	NN	O	O
acetate	NN	O	O
or	NN	O	O
lipopolysaccharide	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	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
proteasome	NN	O	B-protein
inhibitor	NN	O	O
N-Ac-Leu-Leu-norleucinal	NN	O	O
inhibits	NN	O	O
this	NN	O	O
ligand-induced	NN	O	O
degradation	NN	O	O
and	NN	O	O
,	NN	O	O
in	NN	O	O
agreement	NN	O	O
with	NN	O	O
previous	NN	O	O
studies	NN	O	O
,	NN	O	O
stabilizes	NN	O	O
a	NN	O	O
hyperphosphorylated	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

By	NN	O	O
expressing	NN	O	O
mutant	NN	O	O
forms	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
protein	NN	O	O
in	NN	O	O
this	NN	O	O
cell	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
we	NN	O	O
have	NN	O	O
been	NN	O	O
able	NN	O	O
to	NN	O	O
delineate	NN	O	O
the	NN	O	O
sequences	NN	O	O
responsible	NN	O	O
for	NN	O	O
both	NN	O	O
the	NN	O	O
ligand-induced	NN	O	O
phosphorylation	NN	O	O
and	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	NN	O	I-protein
alpha	NN	O	I-protein
.	NN	O	O

Our	NN	O	O
results	NN	O	O
show	NN	O	O
that	NN	O	O
deletion	NN	O	O
of	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
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
molecule	NN	O	O
up	NN	O	O
to	NN	O	O
amino	NN	O	B-protein
acid	NN	O	I-protein
279	NN	O	I-protein
abolishes	NN	O	O
constitutive	NN	O	O
but	NN	O	O
not	NN	O	O
ligand-inducible	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
inhibits	NN	O	O
ligand-inducible	NN	O	O
degradation	NN	O	O
.	NN	O	O

Further	NN	O	O
analysis	NN	O	O
reveals	NN	O	O
that	NN	O	O
the	NN	O	O
inducible	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
maps	NN	O	O
to	NN	O	O
two	NN	O	O
serines	NN	O	O
in	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
protein	NN	O	O
(	NN	O	O
residues	NN	O	O
32	NN	O	O
and	NN	O	O
36	NN	O	O
)	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
mutation	NN	O	O
of	NN	O	O
either	NN	O	O
residue	NN	O	O
is	NN	O	O
sufficient	NN	O	O
to	NN	O	O
abolish	NN	O	O
ligand-induced	NN	O	O
degradation	NN	O	O
,	NN	O	O
whereas	NN	O	O
both	NN	O	O
residues	NN	O	O
must	NN	O	O
be	NN	O	O
mutated	NN	O	O
to	NN	O	O
abolish	NN	O	O
inducible	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
protein	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

Microtubules	NN	O	B-protein
mediate	NN	O	O
cellular	NN	O	O
25-hydroxyvitamin	NN	O	O
D3	NN	O	O
trafficking	NN	O	O
and	NN	O	O
the	NN	O	O
genomic	NN	O	O
response	NN	O	O
to	NN	O	O
1	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
monocytes	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
genomic	NN	O	O
actions	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
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
2D3	NN	O	O
)	NN	O	O
are	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
intracellular	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

Although	NN	O	O
immunocytochemistry	NN	O	O
has	NN	O	O
shown	NN	O	O
that	NN	O	O
disruption	NN	O	O
of	NN	O	O
microtubular	NN	O	O
assembly	NN	O	O
prevents	NN	O	O
nuclear	NN	O	O
access	NN	O	O
of	NN	O	O
the	NN	O	O
sterol-VDR	NN	O	B-protein
complex	NN	O	I-protein
,	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
microtubules	NN	O	B-protein
in	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
has	NN	O	O
not	NN	O	O
been	NN	O	O
studied	NN	O	O
in	NN	O	O
viable	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
examined	NN	O	O
this	NN	O	O
interaction	NN	O	O
in	NN	O	O
normal	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Monocytes	NN	O	O
convert	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
D3	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
to	NN	O	O
24-hydroxylated	NN	O	O
metabolites	NN	O	O
more	NN	O	O
polar	NN	O	O
than	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

Microtubule	NN	O	O
disruption	NN	O	O
totally	NN	O	O
abolished	NN	O	O
the	NN	O	O
ability	NN	O	O
of	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
to	NN	O	O
suppress	NN	O	O
its	NN	O	O
own	NN	O	O
synthesis	NN	O	O
and	NN	O	O
to	NN	O	O
induce	NN	O	O
24-hydroxylase	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
activity	NN	O	O
,	NN	O	O
without	NN	O	O
affecting	NN	O	O
either	NN	O	O
total	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
uptake	NN	O	O
or	NN	O	O
maximal	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
binding	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
intact	NN	O	O
microtubules	NN	O	B-protein
are	NN	O	O
essential	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-dependent	NN	O	O
modulation	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
microtubule	NN	O	B-protein
disruption	NN	O	O
also	NN	O	O
decreased	NN	O	O
monocyte	NN	O	B-cell_type
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
,	NN	O	O
not	NN	O	O
by	NN	O	O
decreasing	NN	O	O
the	NN	O	O
Vmax	NN	O	O
of	NN	O	O
monocyte	NN	O	B-protein
mitochondrial	NN	O	I-protein
1	NN	O	I-protein
alpha-hydroxylase	NN	O	I-protein
but	NN	O	O
through	NN	O	O
an	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
Km	NN	O	O
for	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
examined	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
D3	NN	O	O
transport	NN	O	O
.	NN	O	O

Microtubule	NN	O	O
disruption	NN	O	O
did	NN	O	O
not	NN	O	O
affect	NN	O	O
total	NN	O	O
cellular	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
D3	NN	O	O
uptake	NN	O	O
but	NN	O	O
reduced	NN	O	O
its	NN	O	O
intracellular	NN	O	O
trafficking	NN	O	O
to	NN	O	O
the	NN	O	O
mitochondria	NN	O	O
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
microtubules	NN	O	B-protein
participate	NN	O	O
in	NN	O	O
intracellular	NN	O	O
25	NN	O	O
(	NN	O	O
OH	NN	O	O
)	NN	O	O
D3	NN	O	O
transport	NN	O	O
,	NN	O	O
and	NN	O	O
their	NN	O	O
integrity	NN	O	O
determines	NN	O	O
normal	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
.	NN	O	O

-DOCSTART-	O

Relationship	NN	O	O
between	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
phosphorylation	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
transport	NN	O	O
in	NN	O	O
platelets	NN	O	B-cell_type
:	NN	O	O
a	NN	O	O
new	NN	O	O
approach	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
interrelationship	NN	O	O
between	NN	O	O
the	NN	O	O
two	NN	O	O
messengers	NN	O	O
Ca2+	NN	O	O
and	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
in	NN	O	O
platelet	NN	O	O
function	NN	O	O
is	NN	O	O
well	NN	O	O
documented	NN	O	O
,	NN	O	O
its	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
still	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
established	NN	O	O
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
here	NN	O	O
the	NN	O	O
question	NN	O	O
of	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
platelet	NN	O	B-protein
Ca	NN	O	I-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-ATPases	NN	O	I-protein
by	NN	O	O
cyclic	NN	O	O
AMP	NN	O	O
through	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
using	NN	O	O
a	NN	O	O
pathological	NN	O	O
model	NN	O	O
.	NN	O	O

We	NN	O	O
first	NN	O	O
found	NN	O	O
experimental	NN	O	O
conditions	NN	O	O
where	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-transport	NN	O	O
by	NN	O	O
platelet	NN	O	O
membrane	NN	O	O
vesicles	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
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

Then	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
platelets	NN	O	B-cell_type
of	NN	O	O
patients	NN	O	O
with	NN	O	O
congestive	NN	O	O
heart	NN	O	O
failure	NN	O	O
for	NN	O	O
their	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
potential	NN	O	O
97	NN	O	B-protein
kDa	NN	O	I-protein
Ca	NN	O	I-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-ATPase	NN	O	I-protein
target	NN	O	I-protein
of	NN	O	O
regulation	NN	O	O
through	NN	O	O
the	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
using	NN	O	O
the	NN	O	O
catalytic	NN	O	B-protein
subunit	NN	O	I-protein
of	NN	O	O
the	NN	O	O
cyclic	NN	O	B-protein
AMP-dependent	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
C.	NN	O	B-protein
Sub	NN	O	I-protein
.	NN	O	I-protein
)	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
first	NN	O	O
patients	NN	O	O
studied	NN	O	O
,	NN	O	O
we	NN	O	O
found	NN	O	O
no	NN	O	O
significant	NN	O	O
modification	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
97	NN	O	B-protein
kDa	NN	O	I-protein
Ca	NN	O	I-protein
(	NN	O	I-protein
2+	NN	O	I-protein
)	NN	O	I-protein
-ATPase	NN	O	I-protein
by	NN	O	O
Western	NN	O	O
blotting	NN	O	O
using	NN	O	O
the	NN	O	O
PL/IM	NN	O	B-protein
430	NN	O	I-protein
monoclonal	NN	O	I-protein
antibody	NN	O	I-protein
which	NN	O	O
specifically	NN	O	O
recognized	NN	O	O
this	NN	O	O
isoform	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
the	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
differentially	NN	O	O
phosphorylated	NN	O	O
when	NN	O	O
using	NN	O	O
15	NN	O	O
micrograms/ml	NN	O	O
of	NN	O	O
the	NN	O	O
C.	NN	O	B-protein
Sub	NN	O	I-protein
.	NN	O	I-protein

These	NN	O	O
results	NN	O	O
allowed	NN	O	O
us	NN	O	O
to	NN	O	O
use	NN	O	O
these	NN	O	O
pathological	NN	O	O
platelets	NN	O	B-cell_type
to	NN	O	O
study	NN	O	O
the	NN	O	O
relationship	NN	O	O
between	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
Ca2+	NN	O	O
transport	NN	O	O
by	NN	O	O
selecting	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
severe	NN	O	O
heart	NN	O	O
failure	NN	O	O
.	NN	O	O

We	NN	O	O
could	NN	O	O
show	NN	O	O
a	NN	O	O
decrease	NN	O	O
in	NN	O	O
the	NN	O	O
expression	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
in	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
demonstrate	NN	O	O
a	NN	O	O
lower	NN	O	O
effect	NN	O	O
of	NN	O	O
C.	NN	O	B-protein
Sub	NN	O	I-protein
.	NN	O	I-protein
on	NN	O	O
Ca2+	NN	O	O
transport	NN	O	O
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
by	NN	O	O
studying	NN	O	O
a	NN	O	O
further	NN	O	O
series	NN	O	O
of	NN	O	O
patients	NN	O	O
,	NN	O	O
we	NN	O	O
could	NN	O	O
confirm	NN	O	O
that	NN	O	O
the	NN	O	O
decrease	NN	O	O
in	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
expression	NN	O	O
in	NN	O	O
heart	NN	O	O
failure	NN	O	O
,	NN	O	O
whatever	NN	O	O
its	NN	O	O
extent	NN	O	O
,	NN	O	O
was	NN	O	O
variable	NN	O	O
,	NN	O	O
and	NN	O	O
could	NN	O	O
strictly	NN	O	O
correlate	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
with	NN	O	O
the	NN	O	O
stimulatory	NN	O	O
effect	NN	O	O
of	NN	O	O
C.	NN	O	B-protein
Sub	NN	O	I-protein
.	NN	O	I-protein
on	NN	O	O
Ca2+	NN	O	O
transport	NN	O	O
.	NN	O	O

Besides	NN	O	O
the	NN	O	O
evidence	NN	O	O
for	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
in	NN	O	O
platelets	NN	O	B-cell_type
from	NN	O	O
patients	NN	O	O
with	NN	O	O
heart	NN	O	O
failure	NN	O	O
,	NN	O	O
these	NN	O	O
findings	NN	O	O
constitute	NN	O	O
a	NN	O	O
new	NN	O	O
approach	NN	O	O
in	NN	O	O
favour	NN	O	O
of	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
platelet	NN	O	O
Ca2+	NN	O	O
transport	NN	O	O
through	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
Rap1	NN	O	B-protein
protein	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

An	NN	O	O
IRF-1	NN	O	B-protein
-dependent	NN	O	O
pathway	NN	O	O
of	NN	O	O
DNA	NN	O	O
damage-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
mitogen-activated	NN	O	B-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Lymphocytes	NN	O	B-cell_line
are	NN	O	O
particularly	NN	O	O
susceptible	NN	O	O
to	NN	O	O
DNA	NN	O	O
damage-induced	NN	O	O
apoptosis	NN	O	O
,	NN	O	O
a	NN	O	O
response	NN	O	O
which	NN	O	O
may	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
form	NN	O	O
of	NN	O	O
'altruistic	NN	O	O
suicide	NN	O	O
'	NN	O	O
to	NN	O	O
counter	NN	O	O
their	NN	O	O
intrinsic	NN	O	O
high	NN	O	O
potential	NN	O	O
for	NN	O	O
mutation	NN	O	O
and	NN	O	O
clonal	NN	O	O
expansion	NN	O	O
.	NN	O	O

The	NN	O	O
tumour	NN	O	B-protein
suppressor	NN	O	I-protein
p53	NN	O	I-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
regulate	NN	O	O
this	NN	O	O
type	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
an	NN	O	O
as	NN	O	O
yet	NN	O	O
unknown	NN	O	O
,	NN	O	O
p53	NN	O	B-protein
-independent	NN	O	O
pathway	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
appears	NN	O	O
to	NN	O	O
mediate	NN	O	O
the	NN	O	O
same	NN	O	O
event	NN	O	O
in	NN	O	O
mitogen-activated	NN	O	B-cell_line
mature	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
DNA	NN	O	O
damage-induced	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
these	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
the	NN	O	O
antioncogenic	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
interferon	NN	O	I-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	B-protein
IRF	NN	O	I-protein
)	NN	O	I-protein
-1	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
two	NN	O	O
different	NN	O	O
anti-onco-genic	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
p53	NN	O	B-protein
and	NN	O	O
IRF-1	NN	O	B-protein
,	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
distinct	NN	O	O
apoptotic	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

We	NN	O	O
also	NN	O	O
show	NN	O	O
that	NN	O	O
mitogen	NN	O	O
induction	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-1	NN	O	B-DNA
beta	NN	O	I-DNA
converting	NN	O	I-DNA
enzyme	NN	O	I-DNA
(	NN	O	I-DNA
ICE	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
a	NN	O	O
mammalian	NN	O	B-protein
homologue	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Caenorhabditis	NN	O	B-DNA
elegans	NN	O	I-DNA
cell	NN	O	I-DNA
death	NN	O	I-DNA
gene	NN	O	I-DNA
ced-3	NN	O	B-DNA
,	NN	O	O
is	NN	O	O
IRF-1	NN	O	B-protein
-dependent	NN	O	O
.	NN	O	O

Ectopic	NN	O	O
overexpression	NN	O	O
of	NN	O	O
IRF-1	NN	O	B-protein
results	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
endogenous	NN	O	B-DNA
gene	NN	O	I-DNA
for	NN	O	O
ICE	NN	O	B-protein
and	NN	O	O
enhances	NN	O	O
the	NN	O	O
sensitivity	NN	O	O
of	NN	O	O
cells	NN	O	O
to	NN	O	O
radiation-induced	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Circadian	NN	O	O
rhythm	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
peripheral	NN	O	I-cell_type
leukocytes	NN	O	I-cell_type
and	NN	O	O
their	NN	O	O
reactivity	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

1	NN	O	O
)	NN	O	O
There	NN	O	O
exists	NN	O	O
a	NN	O	O
CR	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
in	NN	O	O
human	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
,	NN	O	O
PMN	NN	O	B-cell_type
,	NN	O	O
and	NN	O	O
monocytes	NN	O	B-cell_type
with	NN	O	O
the	NN	O	O
peak	NN	O	O
values	NN	O	O
from	NN	O	O
0400	NN	O	O
to	NN	O	O
0800	NN	O	O
hr	NN	O	O
and	NN	O	O
the	NN	O	O
trough	NN	O	O
values	NN	O	O
between	NN	O	O
2300	NN	O	O
and	NN	O	O
0000	NN	O	O
hr	NN	O	O
.	NN	O	O

The	NN	O	O
difference	NN	O	O
between	NN	O	O
them	NN	O	O
was	NN	O	O
significant	NN	O	O
statistically	NN	O	O
.	NN	O	O

2	NN	O	O
)	NN	O	O
The	NN	O	O
FI	NN	O	O
of	NN	O	O
the	NN	O	O
chemotactic	NN	O	O
migration	NN	O	O
rate	NN	O	O
of	NN	O	O
PMN	NN	O	B-cell_type
by	NN	O	O
cortisol	NN	O	O
also	NN	O	O
showed	NN	O	O
diurnal	NN	O	O
changes	NN	O	O
which	NN	O	O
were	NN	O	O
synchronous	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
.	NN	O	O

This	NN	O	O
indicates	NN	O	O
that	NN	O	O
the	NN	O	O
CR	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
may	NN	O	O
be	NN	O	O
of	NN	O	O
functional	NN	O	O
significance	NN	O	O
.	NN	O	O

3	NN	O	O
)	NN	O	O
In	NN	O	O
Cushing	NN	O	O
's	NN	O	O
syndrome	NN	O	O
,	NN	O	O
the	NN	O	O
CR	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
was	NN	O	O
normal	NN	O	O
in	NN	O	O
spite	NN	O	O
of	NN	O	O
the	NN	O	O
fact	NN	O	O
that	NN	O	O
the	NN	O	O
CR	NN	O	O
of	NN	O	O
plasma	NN	O	O
cortisol	NN	O	O
was	NN	O	O
disturbed	NN	O	O
.	NN	O	O

This	NN	O	O
indicates	NN	O	O
the	NN	O	O
independency	NN	O	O
of	NN	O	O
the	NN	O	O
CR	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
from	NN	O	O
that	NN	O	O
of	NN	O	O
cortisol	NN	O	O
.	NN	O	O

4	NN	O	O
)	NN	O	O
In	NN	O	O
apoplexy	NN	O	O
caused	NN	O	O
by	NN	O	O
brain	NN	O	O
ischemia	NN	O	O
,	NN	O	O
the	NN	O	O
CR	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
was	NN	O	O
abolished	NN	O	O
in	NN	O	O
patients	NN	O	O
with	NN	O	O
basal	NN	O	O
lesions	NN	O	O
but	NN	O	O
preserved	NN	O	O
when	NN	O	O
the	NN	O	O
lesions	NN	O	O
were	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
cerebral	NN	O	O
cortex	NN	O	O
.	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
main	NN	O	O
``	NN	O	O
circadian	NN	O	O
pacemaker	NN	O	O
''	NN	O	O
of	NN	O	O
GR	NN	O	B-protein
is	NN	O	O
located	NN	O	O
in	NN	O	O
the	NN	O	O
basal	NN	O	O
brain	NN	O	O
,	NN	O	O
most	NN	O	O
probably	NN	O	O
in	NN	O	O
the	NN	O	O
suprachiasmatic	NN	O	O
nuclei	NN	O	O
as	NN	O	O
has	NN	O	O
been	NN	O	O
suggested	NN	O	O
for	NN	O	O
rodents	NN	O	O
.	NN	O	O

-DOCSTART-	O

B-lymphoblastoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
from	NN	O	O
multiple	NN	O	O
sclerosis	NN	O	O
patients	NN	O	O
and	NN	O	O
a	NN	O	O
healthy	NN	O	O
control	NN	O	O
producing	NN	O	O
a	NN	O	O
putative	NN	O	O
new	NN	O	O
human	NN	O	O
retrovirus	NN	O	O
and	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
.	NN	O	O

On	NN	O	O
several	NN	O	O
occasions	NN	O	O
we	NN	O	O
have	NN	O	O
observed	NN	O	O
retrovirus-like	NN	O	O
particles	NN	O	O
(	NN	O	O
RVLPs	NN	O	O
)	NN	O	O
by	NN	O	O
transmission	NN	O	O
electron	NN	O	O
microscopy	NN	O	O
(	NN	O	O
EM	NN	O	O
)	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
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
MS	NN	O	O
.	NN	O	O

Later	NN	O	O
we	NN	O	O
established	NN	O	O
spontaneously	NN	O	O
formed	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
LCLs	NN	O	B-cell_line
)	NN	O	O
from	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
an	NN	O	O
MS-like	NN	O	O
disease	NN	O	O
and	NN	O	O
from	NN	O	O
another	NN	O	O
patient	NN	O	O
with	NN	O	O
MS	NN	O	O
who	NN	O	O
had	NN	O	O
a	NN	O	O
reactivated	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
.	NN	O	O

Both	NN	O	O
LCLs	NN	O	B-cell_line
were	NN	O	O
found	NN	O	O
by	NN	O	O
EM	NN	O	O
to	NN	O	O
produce	NN	O	O
RVLP	NN	O	O
and	NN	O	O
EBV	NN	O	O
particles	NN	O	O
.	NN	O	O

Reverse	NN	O	O
transcriptase	NN	O	O
(	NN	O	O
RT	NN	O	O
)	NN	O	O
assays	NN	O	O
were	NN	O	O
positive	NN	O	O
in	NN	O	O
purified	NN	O	O
viral	NN	O	O
material	NN	O	O
from	NN	O	O
both	NN	O	O
LCLs	NN	O	B-cell_line
.	NN	O	O

To	NN	O	O
substantiate	NN	O	O
these	NN	O	O
findings	NN	O	O
we	NN	O	O
initiated	NN	O	O
an	NN	O	O
intensified	NN	O	O
culturing	NN	O	O
procedure	NN	O	O
and	NN	O	O
were	NN	O	O
able	NN	O	O
to	NN	O	O
establish	NN	O	O
LCLs	NN	O	B-cell_line
from	NN	O	O
5	NN	O	O
out	NN	O	O
of	NN	O	O
21	NN	O	O
consecutive	NN	O	O
MS	NN	O	O
patients	NN	O	O
and	NN	O	O
1	NN	O	O
out	NN	O	O
of	NN	O	O
13	NN	O	O
consecutive	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

All	NN	O	O
LCLs	NN	O	B-cell_line
were	NN	O	O
found	NN	O	O
to	NN	O	O
produce	NN	O	O
both	NN	O	O
RVLP	NN	O	O
and	NN	O	O
EBV	NN	O	O
particles	NN	O	O
by	NN	O	O
EM	NN	O	O
.	NN	O	O

Whether	NN	O	O
the	NN	O	O
putative	NN	O	O
new	NN	O	O
retrovirus	NN	O	O
(	NN	O	O
es	NN	O	O
)	NN	O	O
and	NN	O	O
EBV	NN	O	O
have	NN	O	O
any	NN	O	O
causal	NN	O	O
relationship	NN	O	O
to	NN	O	O
MS	NN	O	O
is	NN	O	O
still	NN	O	O
not	NN	O	O
known	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
findings	NN	O	O
support	NN	O	O
this	NN	O	O
possibility	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
an	NN	O	O
ionomycin/cyclosporin	NN	O	B-DNA
A-responsive	NN	O	I-DNA
element	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
gamma	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Activation	NN	O	O
through	NN	O	O
the	NN	O	O
Ca2+/	NN	O	O
calcineurin	NN	O	B-protein
pathway	NN	O	O
is	NN	O	O
essential	NN	O	O
to	NN	O	O
the	NN	O	O
transcription	NN	O	O
of	NN	O	O
many	NN	O	O
cytokine	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
conserved	NN	O	B-DNA
cis-acting	NN	O	I-DNA
sequence	NN	O	I-DNA
,	NN	O	O
GGAAAA	NN	O	O
,	NN	O	O
and	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
this	NN	O	O
sequence	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
response	NN	O	O
to	NN	O	O
increased	NN	O	O
intracellular	NN	O	O
Ca2+	NN	O	O
concentrations	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
identification	NN	O	O
and	NN	O	O
importance	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
sequence	NN	O	O
in	NN	O	O
a	NN	O	O
non-cytokine	NN	O	B-DNA
gene	NN	O	I-DNA
,	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
gamma	NN	O	I-DNA
(	NN	O	I-DNA
TCRG	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Results	NN	O	O
from	NN	O	O
site-directed	NN	O	O
mutations	NN	O	O
and	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assays	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
this	NN	O	O
sequence	NN	O	O
mediates	NN	O	O
the	NN	O	O
ionomycin-induced	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
TCRG	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Our	NN	O	O
studies	NN	O	O
provide	NN	O	O
an	NN	O	O
explanation	NN	O	O
for	NN	O	O
a	NN	O	O
previous	NN	O	O
observation	NN	O	O
that	NN	O	O
TCRG	NN	O	B-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
for	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
-beta	NN	O	I-protein
,	NN	O	O
are	NN	O	O
increased	NN	O	O
by	NN	O	O
ionomycin	NN	O	O
treatment	NN	O	O
.	NN	O	O

-DOCSTART-	O

Coexpression	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
and	NN	O	O
Sp1	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
in	NN	O	O
human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
1-induced	NN	O	O
,	NN	O	O
dendritic	NN	O	O
cell-T-cell	NN	O	O
syncytia	NN	O	O
.	NN	O	O

Productive	NN	O	O
infection	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	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
typically	NN	O	O
requires	NN	O	O
that	NN	O	O
the	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
be	NN	O	O
stimulated	NN	O	O
with	NN	O	O
antigens	NN	O	O
or	NN	O	O
mitogens	NN	O	O
.	NN	O	O

This	NN	O	O
requirement	NN	O	O
has	NN	O	O
been	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-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
which	NN	O	O
synergizes	NN	O	O
with	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
to	NN	O	O
drive	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Recently	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
found	NN	O	O
that	NN	O	O
vigorous	NN	O	O
replication	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
takes	NN	O	O
place	NN	O	O
in	NN	O	O
nonactivated	NN	O	O
memory	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
after	NN	O	O
syncytium	NN	O	O
formation	NN	O	O
with	NN	O	O
dendritic	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
DCs	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

These	NN	O	O
syncytia	NN	O	O
lack	NN	O	O
activated	NN	O	B-cell_type
cells	NN	O	I-cell_type
as	NN	O	O
determined	NN	O	O
by	NN	O	O
an	NN	O	O
absence	NN	O	O
of	NN	O	O
staining	NN	O	O
for	NN	O	O
Ki-67	NN	O	O
cell	NN	O	O
cycle	NN	O	O
antigen	NN	O	O
.	NN	O	O

The	NN	O	O
expression	NN	O	O
and	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
Sp1	NN	O	B-protein
were	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
analyzed	NN	O	O
in	NN	O	O
isolated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
DCs	NN	O	B-cell_type
from	NN	O	O
humans	NN	O	O
and	NN	O	O
mice	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
used	NN	O	O
immunolabeling	NN	O	O
,	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
,	NN	O	O
and	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
.	NN	O	O

T	NN	O	B-cell_type
cells	NN	O	I-cell_type
lack	NN	O	O
active	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
but	NN	O	O
express	NN	O	O
Sp1	NN	O	B-protein
as	NN	O	O
expected	NN	O	O
.	NN	O	O

DCs	NN	O	B-cell_type
express	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
all	NN	O	O
known	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
and	NN	O	O
Rel	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
with	NN	O	O
activity	NN	O	O
residing	NN	O	O
primarily	NN	O	O
within	NN	O	O
RelB	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

However	NN	O	O
,	NN	O	O
DCs	NN	O	B-cell_type
lack	NN	O	O
Sp1	NN	O	B-protein
,	NN	O	O
which	NN	O	O
may	NN	O	O
explain	NN	O	O
the	NN	O	O
failure	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
to	NN	O	O
replicate	NN	O	O
in	NN	O	O
purified	NN	O	O
DCs	NN	O	B-cell_type
.	NN	O	O

Coexpression	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-protein
occurs	NN	O	O
in	NN	O	O
the	NN	O	O
heterologous	NN	O	B-cell_type
DC-T-cell	NN	O	I-cell_type
syncytia	NN	O	I-cell_type
that	NN	O	O
are	NN	O	O
induced	NN	O	O
by	NN	O	O
HIV-1	NN	O	O
.	NN	O	O

Therefore	NN	O	O
,	NN	O	O
HIV-1-induced	NN	O	O
cell	NN	O	O
fusion	NN	O	O
brings	NN	O	O
together	NN	O	O
factors	NN	O	O
that	NN	O	O
upregulate	NN	O	O
virus	NN	O	O
transcription	NN	O	O
.	NN	O	O

Since	NN	O	O
DCs	NN	O	B-cell_type
and	NN	O	O
memory	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
frequently	NN	O	O
traffic	NN	O	O
together	NN	O	O
in	NN	O	O
situ	NN	O	O
,	NN	O	O
these	NN	O	O
unusual	NN	O	O
heterologous	NN	O	B-cell_type
syncytia	NN	O	I-cell_type
could	NN	O	O
develop	NN	O	O
in	NN	O	O
infected	NN	O	O
individuals	NN	O	O
and	NN	O	O
lead	NN	O	O
to	NN	O	O
chronic	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
without	NN	O	O
ostensible	NN	O	O
immune	NN	O	O
stimulation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cupric	NN	O	O
ion	NN	O	O
blocks	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
through	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
signal-induced	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

A	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
which	NN	O	O
regulates	NN	O	O
expression	NN	O	O
of	NN	O	O
various	NN	O	O
cellular	NN	O	O
genes	NN	O	O
involved	NN	O	O
in	NN	O	O
immune	NN	O	O
responses	NN	O	O
and	NN	O	O
viral	NN	O	B-DNA
genes	NN	O	I-DNA
including	NN	O	O
HIV	NN	O	O
,	NN	O	O
is	NN	O	O
sequestered	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
as	NN	O	O
a	NN	O	O
complex	NN	O	O
with	NN	O	O
an	NN	O	O
inhibitory	NN	O	B-protein
protein	NN	O	I-protein
I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Various	NN	O	O
extracellular	NN	O	O
signals	NN	O	O
induce	NN	O	O
phosphorylation	NN	O	O
and	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	NN	O	I-protein
alpha	NN	O	I-protein
to	NN	O	O
release	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Cu2+	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
inhibit	NN	O	O
the	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
induced	NN	O	O
by	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
TPA	NN	O	O
,	NN	O	O
or	NN	O	O
H2O2	NN	O	O
.	NN	O	O

Deoxycholate	NN	O	O
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	O
extract	NN	O	O
prepared	NN	O	O
from	NN	O	O
cells	NN	O	O
stimulated	NN	O	O
by	NN	O	O
TNF-alpha	NN	O	B-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Cu2+	NN	O	O
resulted	NN	O	O
in	NN	O	O
the	NN	O	O
release	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
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
Cu2+	NN	O	O
interferes	NN	O	O
with	NN	O	O
the	NN	O	O
dissociation	NN	O	O
of	NN	O	O
the	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B-I	NN	O	I-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

Neither	NN	O	O
phosphorylation	NN	O	O
nor	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
was	NN	O	O
observed	NN	O	O
upon	NN	O	O
TNF-alpha	NN	O	B-protein
stimulation	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
Cu2+	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
Cu2+	NN	O	O
inhibits	NN	O	O
the	NN	O	O
release	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
blockade	NN	O	O
of	NN	O	O
a	NN	O	O
signal	NN	O	O
leading	NN	O	O
to	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
.	NN	O	O

-DOCSTART-	O

Cloning	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
from	NN	O	O
human	NN	O	O
NK/T	NN	O	B-cell_type
cells	NN	O	I-cell_type
which	NN	O	O
codes	NN	O	O
for	NN	O	O
a	NN	O	O
protein	NN	O	O
with	NN	O	O
high	NN	O	O
proline	NN	O	O
content	NN	O	O
.	NN	O	O

A	NN	O	O
cDNA	NN	O	B-DNA
clone	NN	O	I-DNA
,	NN	O	O
B4-2	NN	O	B-DNA
,	NN	O	O
was	NN	O	O
isolated	NN	O	O
from	NN	O	O
a	NN	O	O
natural	NN	O	B-DNA
killer	NN	O	I-DNA
(	NN	O	I-DNA
NK	NN	O	I-DNA
)	NN	O	I-DNA
minus	NN	O	I-DNA
T	NN	O	I-DNA
cell	NN	O	I-DNA
subtractive	NN	O	I-DNA
library	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
B4-2	NN	O	B-DNA
clone	NN	O	O
coded	NN	O	O
for	NN	O	O
an	NN	O	O
mRNA	NN	O	B-RNA
of	NN	O	O
2061	NN	O	O
bp	NN	O	O
in	NN	O	O
length	NN	O	O
.	NN	O	O

It	NN	O	O
encodes	NN	O	O
a	NN	O	O
deduced	NN	O	O
327	NN	O	B-protein
aa	NN	O	I-protein
protein	NN	O	I-protein
with	NN	O	O
a	NN	O	O
calculated	NN	O	O
molecular	NN	O	O
mass	NN	O	O
of	NN	O	O
35.2	NN	O	O
kDa	NN	O	O
.	NN	O	O

Searching	NN	O	O
of	NN	O	O
B4-2	NN	O	B-DNA
DNA	NN	O	I-DNA
and	NN	O	O
protein	NN	O	O
sequences	NN	O	O
against	NN	O	O
various	NN	O	O
databases	NN	O	O
revealed	NN	O	O
no	NN	O	O
high	NN	O	O
homology	NN	O	O
to	NN	O	O
other	NN	O	O
sequences	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
B4-2	NN	O	B-DNA
has	NN	O	O
an	NN	O	O
unusually	NN	O	O
high	NN	O	O
proline	NN	O	O
content	NN	O	O
(	NN	O	O
13	NN	O	O
%	NN	O	O
)	NN	O	O
,	NN	O	O
contains	NN	O	O
a	NN	O	O
putative	NN	O	O
nuclear	NN	O	B-protein
targeting	NN	O	I-protein
sequence	NN	O	I-protein
,	NN	O	O
and	NN	O	O
has	NN	O	O
several	NN	O	O
SPXX	NN	O	B-protein
motifs	NN	O	I-protein
which	NN	O	O
are	NN	O	O
frequently	NN	O	O
found	NN	O	O
in	NN	O	O
gene	NN	O	B-protein
regulatory	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

One	NN	O	O
of	NN	O	O
the	NN	O	O
stretches	NN	O	O
of	NN	O	O
prolines	NN	O	O
in	NN	O	O
B4-2	NN	O	B-DNA
closely	NN	O	O
resembles	NN	O	O
the	NN	O	O
ligand	NN	O	O
for	NN	O	O
proteins	NN	O	O
with	NN	O	O
SH3	NN	O	B-protein
domains	NN	O	I-protein
.	NN	O	O

Northern	NN	O	O
hybridization	NN	O	O
data	NN	O	O
showed	NN	O	O
that	NN	O	O
B4-2	NN	O	B-DNA
is	NN	O	O
not	NN	O	O
a	NN	O	O
lymphoid	NN	O	B-DNA
specific	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
a	NN	O	O
hepatoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
also	NN	O	O
weakly	NN	O	O
transcribed	NN	O	O
or	NN	O	O
absent	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
other	NN	O	O
cells	NN	O	O
.	NN	O	O

A	NN	O	O
polyclonal	NN	O	O
antiserum	NN	O	O
raised	NN	O	O
against	NN	O	O
recombinant	NN	O	B-protein
B4-2	NN	O	I-protein
recognizes	NN	O	O
a	NN	O	O
32-34	NN	O	B-protein
kDa	NN	O	I-protein
protein	NN	O	I-protein
in	NN	O	O
lymphocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
JAK1	NN	O	B-protein
,	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
and	NN	O	O
STAT5	NN	O	B-protein
recruitment	NN	O	O
by	NN	O	O
a	NN	O	O
COOH-terminal	NN	O	B-protein
region	NN	O	I-protein
of	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

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
use	NN	O	O
the	NN	O	O
JAK	NN	O	B-protein
-STAT	NN	O	B-protein
pathway	NN	O	O
to	NN	O	O
signal	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
.	NN	O	O

While	NN	O	O
homodimerizing	NN	O	B-protein
cytokine	NN	O	I-protein
receptors	NN	O	I-protein
may	NN	O	O
transmit	NN	O	O
signal	NN	O	O
via	NN	O	O
a	NN	O	O
single	NN	O	O
form	NN	O	O
of	NN	O	O
JAK	NN	O	B-protein
(	NN	O	O
i.e	NN	O	O
.	NN	O	O
growth	NN	O	B-protein
hormone	NN	O	I-protein
receptors	NN	O	I-protein
)	NN	O	O
,	NN	O	O
several	NN	O	O
multicomponent	NN	O	B-protein
cytokine	NN	O	I-protein
receptors	NN	O	I-protein
have	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
require	NN	O	O
simultaneous	NN	O	O
activation	NN	O	O
of	NN	O	O
pairs	NN	O	O
of	NN	O	O
different	NN	O	O
JAK	NN	O	B-protein
kinases	NN	O	I-protein
(	NN	O	O
i.e	NN	O	O
.	NN	O	O
interferon	NN	O	B-protein
receptors	NN	O	I-protein
)	NN	O	O
.	NN	O	O

Recent	NN	O	O
evidence	NN	O	O
for	NN	O	O
a	NN	O	O
preferential	NN	O	O
coupling	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
to	NN	O	O
interleukin-2	NN	O	B-protein
receptor-gamma	NN	O	I-protein
(	NN	O	O
IL-2R	NN	O	B-protein
gamma	NN	O	I-protein
)	NN	O	O
and	NN	O	O
JAK1	NN	O	B-protein
to	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
supports	NN	O	O
the	NN	O	O
concept	NN	O	O
of	NN	O	O
heterotrans-activation	NN	O	O
of	NN	O	O
JAK1	NN	O	B-protein
and	NN	O	O
JAK3	NN	O	B-protein
caused	NN	O	O
by	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
heterodimerization	NN	O	O
of	NN	O	O
their	NN	O	O
receptor	NN	O	O
partners	NN	O	O
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
verified	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
to	NN	O	O
cause	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
and	NN	O	O
JAK3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
stimulated	NN	O	O
JAK3	NN	O	B-protein
to	NN	O	O
a	NN	O	O
significantly	NN	O	O
larger	NN	O	O
extent	NN	O	O
than	NN	O	O
JAK1	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
and	NN	O	O
the	NN	O	O
YT	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
conclusion	NN	O	O
was	NN	O	O
based	NN	O	O
upon	NN	O	O
several	NN	O	O
independent	NN	O	O
criteria	NN	O	O
,	NN	O	O
including	NN	O	O
more	NN	O	O
vigorous	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
,	NN	O	O
more	NN	O	O
marked	NN	O	O
enzymatic	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
higher	NN	O	O
abundance	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
in	NN	O	O
activated	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
when	NN	O	O
human	NN	O	B-protein
IL-2R	NN	O	I-protein
beta	NN	O	I-protein
was	NN	O	O
stably	NN	O	O
expressed	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
,	NN	O	O
robust	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
proliferation	NN	O	O
and	NN	O	O
JAK3	NN	O	B-protein
activation	NN	O	O
occurred	NN	O	O
without	NN	O	O
detectable	NN	O	O
involvement	NN	O	O
of	NN	O	O
either	NN	O	O
JAK1	NN	O	B-protein
,	NN	O	O
JAK2	NN	O	B-protein
or	NN	O	O
TYK2	NN	O	B-protein
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
propose	NN	O	O
that	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
signal	NN	O	O
transduction	NN	O	O
does	NN	O	O
not	NN	O	O
depend	NN	O	O
on	NN	O	O
equimolar	NN	O	O
heterodimerization	NN	O	O
of	NN	O	O
JAK1	NN	O	B-protein
and	NN	O	O
JAK3	NN	O	B-protein
following	NN	O	O
IL-2	NN	O	B-protein
-induced	NN	O	O
heterodimerization	NN	O	O
of	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

Nonetheless	NN	O	O
,	NN	O	O
a	NN	O	O
membrane-proximal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
human	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
(	NN	O	O
Asn240-Leu335	NN	O	B-protein
)	NN	O	O
was	NN	O	O
critical	NN	O	O
for	NN	O	O
JAK3	NN	O	B-protein
activation	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
present	NN	O	O
in	NN	O	O
activated	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	I-protein
complexes	NN	O	I-protein
increased	NN	O	O
with	NN	O	O
time	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
stabilization	NN	O	O
of	NN	O	O
JAK3	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
the	NN	O	O
receptor	NN	O	B-protein
complex	NN	O	I-protein
relies	NN	O	O
on	NN	O	O
both	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

Moreover	NN	O	O
,	NN	O	O
STAT5	NN	O	B-protein
was	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
the	NN	O	O
predominant	NN	O	O
STAT	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
used	NN	O	O
by	NN	O	O
IL-2	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
and	NN	O	O
specifically	NN	O	O
required	NN	O	O
a	NN	O	O
COOH-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
IL-2R	NN	O	B-protein
beta	NN	O	I-protein
(	NN	O	O
Ser386-Val525	NN	O	B-protein
)	NN	O	O
,	NN	O	O
while	NN	O	O
STAT5	NN	O	B-protein
recruitment	NN	O	O
was	NN	O	O
not	NN	O	O
correlated	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
IL-2R	NN	O	B-protein
gamma	NN	O	I-protein
or	NN	O	O
JAK3	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Up-regulation	NN	O	O
of	NN	O	O
high-affinity	NN	O	O
dehydroepiandrosterone	NN	O	O
binding	NN	O	O
activity	NN	O	O
by	NN	O	O
dehydroepiandrosterone	NN	O	O
in	NN	O	O
activated	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Although	NN	O	O
evidence	NN	O	O
indicates	NN	O	O
that	NN	O	O
dehydroepiandrosterone	NN	O	O
(	NN	O	O
DHEA	NN	O	O
)	NN	O	O
exerts	NN	O	O
direct	NN	O	O
physiological	NN	O	O
effects	NN	O	O
,	NN	O	O
its	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	NN	O	O
remains	NN	O	O
unknown	NN	O	O
.	NN	O	O

DHEA	NN	O	O
binding	NN	O	O
sites	NN	O	O
were	NN	O	O
examined	NN	O	O
using	NN	O	O
a	NN	O	O
whole-cell	NN	O	O
binding	NN	O	O
assay	NN	O	O
in	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
revealing	NN	O	O
that	NN	O	O
a	NN	O	O
single	NN	O	O
class	NN	O	O
of	NN	O	O
high-affinity	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
DHEA	NN	O	O
(	NN	O	O
dissociation	NN	O	O
constant	NN	O	O
=	NN	O	O
7.4	NN	O	O
+/-	NN	O	O
0.53	NN	O	O
nmol/L	NN	O	O
,	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SE	NN	O	O
,	NN	O	O
n	NN	O	O
=	NN	O	O
4	NN	O	O
)	NN	O	O
was	NN	O	O
greatly	NN	O	O
increased	NN	O	O
when	NN	O	O
treated	NN	O	O
with	NN	O	O
DHEA	NN	O	O
,	NN	O	O
phorbol-12-myristate-13-acetate	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
Ca2+	NN	O	O
ionophore	NN	O	O
A23187	NN	O	O
.	NN	O	O

Bound	NN	O	O
[	NN	O	O
3H	NN	O	O
]	NN	O	O
DHEA	NN	O	O
was	NN	O	O
displaced	NN	O	O
sensitively	NN	O	O
by	NN	O	O
DHEA	NN	O	O
and	NN	O	O
secondarily	NN	O	O
by	NN	O	O
dihydrotestosterone	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
effectively	NN	O	O
by	NN	O	O
other	NN	O	O
steroids	NN	O	O
,	NN	O	O
including	NN	O	O
DHEA	NN	O	O
sulfate	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
not	NN	O	O
only	NN	O	O
indicate	NN	O	O
the	NN	O	O
existence	NN	O	O
of	NN	O	O
a	NN	O	O
DHEA	NN	O	B-protein
receptor	NN	O	I-protein
,	NN	O	O
but	NN	O	O
also	NN	O	O
suggest	NN	O	O
that	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
become	NN	O	O
susceptible	NN	O	O
to	NN	O	O
regulation	NN	O	O
by	NN	O	O
DHEA	NN	O	O
during	NN	O	O
the	NN	O	O
process	NN	O	O
of	NN	O	O
signal-induced	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Ubiquitin-mediated	NN	O	O
processing	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
transcriptional	NN	O	B-protein
activator	NN	O	I-protein
precursor	NN	O	I-protein
p105	NN	O	B-protein
.	NN	O	O

Reconstitution	NN	O	O
of	NN	O	O
a	NN	O	O
cell-free	NN	O	O
system	NN	O	O
and	NN	O	O
identification	NN	O	O
of	NN	O	O
the	NN	O	O
ubiquitin-carrier	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
E2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
novel	NN	O	O
ubiquitin-protein	NN	O	B-protein
ligase	NN	O	I-protein
,	NN	O	O
E3	NN	O	B-protein
,	NN	O	O
involved	NN	O	O
in	NN	O	O
conjugation	NN	O	O
.	NN	O	O

In	NN	O	O
most	NN	O	O
cases	NN	O	O
,	NN	O	O
the	NN	O	O
transcriptional	NN	O	B-protein
factor	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
is	NN	O	O
a	NN	O	O
heterodimer	NN	O	B-protein
consisting	NN	O	O
of	NN	O	O
two	NN	O	O
subunits	NN	O	O
,	NN	O	O
p50	NN	O	B-protein
and	NN	O	O
p65	NN	O	B-protein
,	NN	O	O
which	NN	O	O
are	NN	O	O
encoded	NN	O	O
by	NN	O	O
two	NN	O	O
distinct	NN	O	O
genes	NN	O	O
of	NN	O	O
the	NN	O	O
Rel	NN	O	B-DNA
family	NN	O	I-DNA
.	NN	O	O

p50	NN	O	B-protein
is	NN	O	O
translated	NN	O	O
as	NN	O	O
a	NN	O	O
precursor	NN	O	O
of	NN	O	O
105	NN	O	B-protein
kDa	NN	O	I-protein
.	NN	O	O

The	NN	O	O
C-terminal	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
the	NN	O	O
precursor	NN	O	O
is	NN	O	O
rapidly	NN	O	O
degraded	NN	O	O
,	NN	O	O
forming	NN	O	O
the	NN	O	O
mature	NN	O	B-protein
p50	NN	O	I-protein
subunit	NN	O	I-protein
consisted	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
molecule	NN	O	O
.	NN	O	O

The	NN	O	O
mechanism	NN	O	O
of	NN	O	O
generation	NN	O	O
of	NN	O	O
p50	NN	O	B-protein
is	NN	O	O
not	NN	O	O
known	NN	O	O
.	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
ubiquitin	NN	O	B-protein
-proteasome	NN	O	B-protein
system	NN	O	O
is	NN	O	O
involved	NN	O	O
in	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
specific	NN	O	O
enzymes	NN	O	B-protein
involved	NN	O	O
and	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
limited	NN	O	O
proteolysis	NN	O	O
,	NN	O	O
in	NN	O	O
which	NN	O	O
half	NN	O	O
of	NN	O	O
the	NN	O	O
molecule	NN	O	O
is	NN	O	O
spared	NN	O	O
,	NN	O	O
have	NN	O	O
been	NN	O	O
obscure	NN	O	O
.	NN	O	O

Palombella	NN	O	O
and	NN	O	O
colleagues	NN	O	O
(	NN	O	O
Palombella	NN	O	O
,	NN	O	O
V.J.	NN	O	O
,	NN	O	O
Rando	NN	O	O
,	NN	O	O
O.J.	NN	O	O
,	NN	O	O
Goldberg	NN	O	O
,	NN	O	O
A.L.	NN	O	O
,	NN	O	O
and	NN	O	O
Maniatis	NN	O	O
,	NN	O	O
T.	NN	O	O
(	NN	O	O
1994	NN	O	O
)	NN	O	O
Cell	NN	O	O
78	NN	O	O
,	NN	O	O
773-785	NN	O	O
)	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
ubiquitin	NN	O	B-protein
is	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
processing	NN	O	O
in	NN	O	O
a	NN	O	O
cell-free	NN	O	O
system	NN	O	O
of	NN	O	O
a	NN	O	O
truncated	NN	O	O
,	NN	O	O
artificially	NN	O	O
constructed	NN	O	O
,	NN	O	O
60-kDa	NN	O	B-protein
precursor	NN	O	I-protein
.	NN	O	O

They	NN	O	O
have	NN	O	O
also	NN	O	O
shown	NN	O	O
that	NN	O	O
proteasome	NN	O	B-protein
inhibitors	NN	O	O
block	NN	O	O
the	NN	O	O
processing	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
this	NN	O	O
study	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
reconstitution	NN	O	O
of	NN	O	O
a	NN	O	O
cell-free	NN	O	O
processing	NN	O	O
system	NN	O	O
and	NN	O	O
demonstrate	NN	O	O
directly	NN	O	O
that	NN	O	O
:	NN	O	O
(	NN	O	O
a	NN	O	O
)	NN	O	O
the	NN	O	O
ubiquitin	NN	O	B-protein
-proteasome	NN	O	B-protein
system	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
processing	NN	O	O
of	NN	O	O
the	NN	O	O
intact	NN	O	O
p105	NN	O	B-protein
precursor	NN	O	O
,	NN	O	O
(	NN	O	O
b	NN	O	O
)	NN	O	O
conjugation	NN	O	O
of	NN	O	O
ubiquitin	NN	O	B-protein
to	NN	O	O
the	NN	O	O
precursor	NN	O	O
is	NN	O	O
an	NN	O	O
essential	NN	O	O
intermediate	NN	O	O
step	NN	O	O
in	NN	O	O
the	NN	O	O
processing	NN	O	O
,	NN	O	O
(	NN	O	O
c	NN	O	O
)	NN	O	O
the	NN	O	O
recently	NN	O	O
discovered	NN	O	O
novel	NN	O	O
species	NN	O	O
of	NN	O	O
the	NN	O	O
ubiquitin-carrier	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
E2-F1	NN	O	B-protein
,	NN	O	O
that	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
conjugation	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
p53	NN	O	B-protein
,	NN	O	O
is	NN	O	O
also	NN	O	O
required	NN	O	O
for	NN	O	O
the	NN	O	O
limited	NN	O	O
processing	NN	O	O
of	NN	O	O
the	NN	O	O
p105	NN	O	B-protein
precursor	NN	O	I-protein
,	NN	O	O
and	NN	O	O
(	NN	O	O
d	NN	O	O
)	NN	O	O
a	NN	O	O
novel	NN	O	O
,	NN	O	O
approximately	NN	O	O
320-kDa	NN	O	B-protein
species	NN	O	I-protein
of	NN	O	O
ubiquitin-protein	NN	O	B-protein
ligase	NN	O	I-protein
,	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
process	NN	O	O
.	NN	O	O

This	NN	O	O
novel	NN	O	O
enzyme	NN	O	O
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
E6-AP	NN	O	B-protein
,	NN	O	O
the	NN	O	O
p53-conjugating	NN	O	B-protein
ligase	NN	O	I-protein
,	NN	O	O
and	NN	O	O
from	NN	O	O
E3	NN	O	B-protein
alpha	NN	O	I-protein
,	NN	O	O
the	NN	O	O
``	NN	O	B-protein
N-end	NN	O	I-protein
rule	NN	O	I-protein
''	NN	O	I-protein
ligase	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Flutamide	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
hirsutism	NN	O	O
:	NN	O	O
long-term	NN	O	O
clinical	NN	O	O
effects	NN	O	O
,	NN	O	O
endocrine	NN	O	O
changes	NN	O	O
,	NN	O	O
and	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
behavior	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
long-term	NN	O	O
effects	NN	O	O
of	NN	O	O
treatment	NN	O	O
with	NN	O	O
low	NN	O	O
doses	NN	O	O
of	NN	O	O
flutamide	NN	O	O
on	NN	O	O
clinical	NN	O	O
and	NN	O	O
hormonal	NN	O	O
parameters	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
on	NN	O	O
the	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
status	NN	O	O
,	NN	O	O
in	NN	O	O
hirsute	NN	O	O
women	NN	O	O
.	NN	O	O

DESIGN	NN	O	O
:	NN	O	O
Eighteen	NN	O	O
hirsute	NN	O	O
patients	NN	O	O
with	NN	O	O
regular	NN	O	O
menses	NN	O	O
were	NN	O	O
studied	NN	O	O
basally	NN	O	O
and	NN	O	O
during	NN	O	O
treatment	NN	O	O
with	NN	O	O
125	NN	O	O
mg	NN	O	O
flutamide	NN	O	O
,	NN	O	O
three	NN	O	O
times	NN	O	O
per	NN	O	O
day	NN	O	O
for	NN	O	O
12	NN	O	O
months	NN	O	O
.	NN	O	O

Barrier	NN	O	O
or	NN	O	O
intrauterine	NN	O	O
contraception	NN	O	O
was	NN	O	O
used	NN	O	O
during	NN	O	O
the	NN	O	O
study	NN	O	O
in	NN	O	O
sexually	NN	O	O
active	NN	O	O
women	NN	O	O
.	NN	O	O

Safety	NN	O	O
parameters	NN	O	O
were	NN	O	O
assessed	NN	O	O
throughout	NN	O	O
the	NN	O	O
study	NN	O	O
.	NN	O	O

Hirsutism	NN	O	O
,	NN	O	O
graded	NN	O	O
by	NN	O	O
the	NN	O	O
modified	NN	O	O
Ferriman-Gallwey	NN	O	O
score	NN	O	O
,	NN	O	O
and	NN	O	O
hormonal	NN	O	O
parameters	NN	O	O
were	NN	O	O
evaluated	NN	O	O
basally	NN	O	O
and	NN	O	O
at	NN	O	O
4-month	NN	O	O
intervals	NN	O	O
during	NN	O	O
treatment	NN	O	O
.	NN	O	O

Gonadotropin-releasing	NN	O	O
hormone	NN	O	O
and	NN	O	O
ACTH	NN	O	O
stimulation	NN	O	O
tests	NN	O	O
were	NN	O	O
performed	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
3	NN	O	O
to	NN	O	O
4	NN	O	O
months	NN	O	O
of	NN	O	O
therapy	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
androgen	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
measured	NN	O	O
,	NN	O	O
in	NN	O	O
both	NN	O	O
the	NN	O	O
follicular	NN	O	O
and	NN	O	O
luteal	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
basally	NN	O	O
and	NN	O	O
after	NN	O	O
4	NN	O	O
months	NN	O	O
of	NN	O	O
flutamide	NN	O	O
treatment	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Flutamide	NN	O	O
was	NN	O	O
well	NN	O	O
tolerated	NN	O	O
in	NN	O	O
all	NN	O	O
women	NN	O	O
,	NN	O	O
with	NN	O	O
the	NN	O	O
noticeable	NN	O	O
exception	NN	O	O
of	NN	O	O
one	NN	O	O
patient	NN	O	O
who	NN	O	O
presented	NN	O	O
increased	NN	O	O
serum	NN	O	B-protein
transaminase	NN	O	I-protein
after	NN	O	O
8	NN	O	O
months	NN	O	O
of	NN	O	O
therapy	NN	O	O
.	NN	O	O

Hirsutism	NN	O	O
markedly	NN	O	O
improved	NN	O	O
in	NN	O	O
all	NN	O	O
women	NN	O	O
during	NN	O	O
the	NN	O	O
treatment	NN	O	O
(	NN	O	O
Ferriman-Gallwey	NN	O	O
score	NN	O	O
after	NN	O	O
1	NN	O	O
year	NN	O	O
:	NN	O	O
4.1	NN	O	O
+/-	NN	O	O
0.5	NN	O	O
versus	NN	O	O
14.1	NN	O	O
+/-	NN	O	O
0.9	NN	O	O
)	NN	O	O
.	NN	O	O

A	NN	O	O
reduction	NN	O	O
of	NN	O	O
serum	NN	O	O
androgens	NN	O	O
was	NN	O	O
found	NN	O	O
,	NN	O	O
whereas	NN	O	O
no	NN	O	O
change	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
either	NN	O	O
basal	NN	O	O
or	NN	O	O
GnRH-stimulated	NN	O	O
gonadotropins	NN	O	O
or	NN	O	O
in	NN	O	O
the	NN	O	O
cortisol	NN	O	O
and	NN	O	O
17	NN	O	O
alpha-hydroxyprogesterone	NN	O	O
response	NN	O	O
to	NN	O	O
ACTH	NN	O	O
.	NN	O	O

Cycles	NN	O	O
remained	NN	O	O
ovulatory	NN	O	O
.	NN	O	O

Before	NN	O	O
treatment	NN	O	O
,	NN	O	O
the	NN	O	O
number	NN	O	O
of	NN	O	O
androgen	NN	O	B-protein
receptors	NN	O	I-protein
was	NN	O	O
higher	NN	O	O
in	NN	O	O
the	NN	O	O
luteal	NN	O	O
than	NN	O	O
in	NN	O	O
the	NN	O	O
follicular	NN	O	O
phase	NN	O	O
.	NN	O	O

This	NN	O	O
rhythmic	NN	O	O
differentiation	NN	O	O
disappeared	NN	O	O
after	NN	O	O
the	NN	O	O
patients	NN	O	O
had	NN	O	O
been	NN	O	O
given	NN	O	O
the	NN	O	O
antiandrogen	NN	O	O
drug	NN	O	O
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Flutamide	NN	O	O
is	NN	O	O
effective	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
hirsutism	NN	O	O
but	NN	O	O
requires	NN	O	O
constant	NN	O	O
surveillance	NN	O	O
of	NN	O	O
liver	NN	O	O
function	NN	O	O
.	NN	O	O

Androgen	NN	O	O
receptor	NN	O	O
blockade	NN	O	O
might	NN	O	O
be	NN	O	O
potentiated	NN	O	O
by	NN	O	O
a	NN	O	O
reduction	NN	O	O
of	NN	O	O
serum	NN	O	O
androgens	NN	O	O
.	NN	O	O

Flutamide	NN	O	O
affects	NN	O	O
androgen	NN	O	B-protein
receptor	NN	O	I-protein
behavior	NN	O	O
during	NN	O	O
the	NN	O	O
menstrual	NN	O	O
cycle	NN	O	O
.	NN	O	O

The	NN	O	O
meaning	NN	O	O
of	NN	O	O
this	NN	O	O
finding	NN	O	O
remains	NN	O	O
to	NN	O	O
be	NN	O	O
elucidated	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
expression	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-protein
tat	NN	O	I-protein
protein	NN	O	I-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
using	NN	O	O
a	NN	O	O
BK	NN	O	O
virus	NN	O	O
vector	NN	O	O
.	NN	O	O

The	NN	O	O
production	NN	O	O
and	NN	O	O
characterization	NN	O	O
of	NN	O	O
Jurkat	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
that	NN	O	O
constitutively	NN	O	O
express	NN	O	O
functional	NN	O	O
human	NN	O	B-protein
immune	NN	O	I-protein
deficiency	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
,	NN	O	O
using	NN	O	O
a	NN	O	O
BK	NN	O	O
virus	NN	O	O
plasmid	NN	O	O
expression	NN	O	O
vector	NN	O	O
and	NN	O	O
HIV-1	NN	O	B-DNA
tat	NN	O	I-DNA
cDNA	NN	O	I-DNA
,	NN	O	O
is	NN	O	O
described	NN	O	O
.	NN	O	O

An	NN	O	O
increased	NN	O	O
growth	NN	O	O
rate	NN	O	O
of	NN	O	O
these	NN	O	O
Jurkat-tat	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
as	NN	O	O
compared	NN	O	O
with	NN	O	O
control	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
was	NN	O	O
observed	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
PEBP2	NN	O	B-protein
alpha/AML-1-related	NN	O	I-protein
factor	NN	O	I-protein
increases	NN	O	O
osteocalcin	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
through	NN	O	O
its	NN	O	O
binding	NN	O	O
to	NN	O	O
an	NN	O	O
osteoblast-specific	NN	O	B-DNA
cis-acting	NN	O	I-DNA
element	NN	O	I-DNA
.	NN	O	O

To	NN	O	O
identify	NN	O	O
osteoblast-specific	NN	O	B-DNA
cis-acting	NN	O	I-DNA
elements	NN	O	I-DNA
and	NN	O	O
trans-acting	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
we	NN	O	O
initiated	NN	O	O
an	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
of	NN	O	O
a	NN	O	O
mouse	NN	O	B-DNA
osteocalcin	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
an	NN	O	O
osteoblast-specific	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
this	NN	O	O
promoter	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
two	NN	O	O
osteoblast-specific	NN	O	B-DNA
cis-acting	NN	O	I-DNA
elements	NN	O	I-DNA
(	NN	O	O
Ducy	NN	O	O
,	NN	O	O
P.and	NN	O	O
Karsenty	NN	O	O
,	NN	O	O
G.	NN	O	O
(	NN	O	O
1995	NN	O	O
)	NN	O	O
Mol.Cell.Biol.15	NN	O	O
,	NN	O	O
1858-1869	NN	O	O
)	NN	O	O
.	NN	O	O

The	NN	O	O
sequence	NN	O	O
of	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
elements	NN	O	B-DNA
,	NN	O	O
OSE2	NN	O	O
,	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-DNA
site	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
PEBP2	NN	O	B-protein
alpha/AML-1	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
the	NN	O	O
mammalian	NN	O	B-protein
homologues	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Drosophila	NN	O	B-protein
Runt	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
show	NN	O	O
,	NN	O	O
using	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
,	NN	O	O
recombinant	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
a	NN	O	O
specific	NN	O	O
antiserum	NN	O	O
against	NN	O	O
AML-1	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
DNA-binding	NN	O	O
assays	NN	O	O
,	NN	O	O
that	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
AML-1B	NN	O	B-protein
,	NN	O	O
binds	NN	O	O
specifically	NN	O	O
to	NN	O	O
OSE2	NN	O	O
and	NN	O	O
is	NN	O	O
immunologically	NN	O	O
related	NN	O	O
to	NN	O	O
OSF2	NN	O	B-protein
,	NN	O	O
the	NN	O	O
factor	NN	O	O
present	NN	O	O
in	NN	O	O
osteoblast	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
OSE2	NN	O	O
.	NN	O	O

By	NN	O	O
DNA	NN	O	O
cotransfection	NN	O	O
experiments	NN	O	O
,	NN	O	O
we	NN	O	O
also	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
AML-1B	NN	O	B-protein
can	NN	O	O
increase	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
short	NN	O	O
osteocalcin	NN	O	B-DNA
promoter	NN	O	I-DNA
through	NN	O	O
its	NN	O	O
binding	NN	O	O
to	NN	O	O
OSE2	NN	O	O
.	NN	O	O

Lastly	NN	O	O
,	NN	O	O
the	NN	O	O
different	NN	O	O
mobilities	NN	O	O
of	NN	O	O
osteoblast	NN	O	B-protein
nuclear	NN	O	I-protein
extract-DNA	NN	O	I-protein
complexes	NN	O	I-protein
compared	NN	O	O
with	NN	O	O
T-cell	NN	O	B-protein
nuclear	NN	O	I-protein
extract-DNA	NN	O	I-protein
complexes	NN	O	I-protein
,	NN	O	O
along	NN	O	O
with	NN	O	O
the	NN	O	O
inability	NN	O	O
of	NN	O	O
OSF2	NN	O	B-protein
to	NN	O	O
be	NN	O	O
upregulated	NN	O	O
by	NN	O	O
retinoic	NN	O	O
acid	NN	O	O
,	NN	O	O
unlike	NN	O	O
the	NN	O	O
other	NN	O	O
PEBP2	NN	O	B-protein
alpha	NN	O	I-protein
factors	NN	O	I-protein
,	NN	O	O
suggest	NN	O	O
that	NN	O	O
OSF2	NN	O	B-protein
is	NN	O	O
a	NN	O	O
new	NN	O	O
member	NN	O	O
of	NN	O	O
this	NN	O	O
family	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
this	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
AML-1B	NN	O	B-protein
can	NN	O	O
increase	NN	O	O
gene	NN	O	O
expression	NN	O	O
of	NN	O	O
an	NN	O	O
osteoblast-specific	NN	O	B-DNA
gene	NN	O	I-DNA
through	NN	O	O
its	NN	O	O
binding	NN	O	O
to	NN	O	O
an	NN	O	O
osteoblast-specific	NN	O	B-DNA
cis-acting	NN	O	I-DNA
element	NN	O	I-DNA
and	NN	O	O
presents	NN	O	O
evidence	NN	O	O
that	NN	O	O
OSF2	NN	O	B-protein
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
PEBP2	NN	O	B-protein
alpha/AML-1	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

-DOCSTART-	O

Initiation	NN	O	B-protein
binding	NN	O	I-protein
repressor	NN	O	I-protein
,	NN	O	O
a	NN	O	O
factor	NN	O	O
that	NN	O	O
binds	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
of	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
is	NN	O	O
a	NN	O	O
glycosylated	NN	O	B-protein
member	NN	O	I-protein
of	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
cell	NN	O	B-protein
growth	NN	O	I-protein
regulators	NN	O	I-protein
[	NN	O	O
corrected	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
Mol	NN	O	O
Cell	NN	O	O
Biol	NN	O	O
1996	NN	O	O
Feb	NN	O	O
;	NN	O	O
16	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
:	NN	O	O
735	NN	O	O
]	NN	O	O

Initiation	NN	O	B-protein
binding	NN	O	I-protein
repressor	NN	O	I-protein
[	NN	O	O
corrected	NN	O	O
]	NN	O	O
(	NN	O	O
IBR	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
chicken	NN	O	B-protein
erythrocyte	NN	O	I-protein
factor	NN	O	I-protein
(	NN	O	O
apparent	NN	O	O
molecular	NN	O	O
mass	NN	O	O
,	NN	O	O
70	NN	O	O
to	NN	O	O
73	NN	O	O
kDa	NN	O	O
)	NN	O	O
that	NN	O	O
binds	NN	O	O
to	NN	O	O
the	NN	O	O
sequences	NN	O	O
spanning	NN	O	O
the	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
of	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
repressing	NN	O	O
its	NN	O	O
transcription	NN	O	O
.	NN	O	O

A	NN	O	O
variety	NN	O	O
of	NN	O	O
other	NN	O	O
cells	NN	O	O
,	NN	O	O
including	NN	O	O
transformed	NN	O	B-cell_type
erythroid	NN	O	I-cell_type
precursors	NN	O	I-cell_type
,	NN	O	O
do	NN	O	O
not	NN	O	O
have	NN	O	O
IBR	NN	O	B-protein
but	NN	O	O
a	NN	O	O
factor	NN	O	O
referred	NN	O	O
to	NN	O	O
as	NN	O	O
IBF	NN	O	B-protein
(	NN	O	O
68	NN	O	O
to	NN	O	O
70	NN	O	O
kDa	NN	O	O
)	NN	O	O
that	NN	O	O
recognizes	NN	O	O
the	NN	O	O
same	NN	O	O
IBR	NN	O	B-protein
sites	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
cloned	NN	O	O
the	NN	O	O
IBR	NN	O	B-DNA
cDNA	NN	O	I-DNA
and	NN	O	O
studied	NN	O	O
the	NN	O	O
relationship	NN	O	O
of	NN	O	O
IBR	NN	O	B-protein
and	NN	O	O
IBF	NN	O	B-protein
.	NN	O	O

IBR	NN	O	B-protein
is	NN	O	O
a	NN	O	O
503-amino-acid-long	NN	O	B-protein
acidic	NN	O	I-protein
protein	NN	O	I-protein
which	NN	O	O
is	NN	O	O
99.0	NN	O	O
%	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
recently	NN	O	O
reported	NN	O	O
human	NN	O	B-protein
NRF-1/alpha-Pal	NN	O	I-protein
factor	NN	O	I-protein
and	NN	O	O
highly	NN	O	O
related	NN	O	O
to	NN	O	O
the	NN	O	O
invertebrate	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
P3A2	NN	O	B-protein
and	NN	O	O
erected	NN	O	B-protein
wing	NN	O	I-protein
gene	NN	O	I-protein
product	NN	O	I-protein
(	NN	O	O
EWG	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
present	NN	O	O
evidence	NN	O	O
that	NN	O	O
IBR	NN	O	B-protein
and	NN	O	O
IBF	NN	O	B-protein
are	NN	O	O
most	NN	O	O
likely	NN	O	O
identical	NN	O	O
proteins	NN	O	O
,	NN	O	O
differing	NN	O	O
in	NN	O	O
their	NN	O	O
degree	NN	O	O
of	NN	O	O
glycosylation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
several	NN	O	O
molecular	NN	O	O
aspects	NN	O	O
of	NN	O	O
IBR/F	NN	O	B-protein
and	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
factor	NN	O	O
associates	NN	O	O
as	NN	O	O
stable	NN	O	O
homodimers	NN	O	B-protein
and	NN	O	O
that	NN	O	O
the	NN	O	O
dimer	NN	O	B-protein
is	NN	O	O
the	NN	O	O
relevant	NN	O	B-protein
DNA-binding	NN	O	I-protein
species	NN	O	I-protein
.	NN	O	O

The	NN	O	O
evolutionarily	NN	O	O
conserved	NN	O	O
N-terminal	NN	O	B-protein
half	NN	O	I-protein
of	NN	O	O
IBR/F	NN	O	B-protein
harbors	NN	O	O
the	NN	O	O
DNA-binding/dimerization	NN	O	B-protein
domain	NN	O	I-protein
(	NN	O	O
outer	NN	O	O
limits	NN	O	O
,	NN	O	O
127	NN	O	O
to	NN	O	O
283	NN	O	O
)	NN	O	O
,	NN	O	O
one	NN	O	O
or	NN	O	O
several	NN	O	O
casein	NN	O	B-protein
kinase	NN	O	I-protein
II	NN	O	I-protein
sites	NN	O	I-protein
(	NN	O	O
37	NN	O	O
to	NN	O	O
67	NN	O	O
)	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
bipartite	NN	O	B-protein
nuclear	NN	O	I-protein
localization	NN	O	I-protein
signal	NN	O	I-protein
(	NN	O	O
89	NN	O	O
to	NN	O	O
106	NN	O	O
)	NN	O	O
which	NN	O	O
appears	NN	O	O
to	NN	O	O
be	NN	O	O
necessary	NN	O	O
for	NN	O	O
nuclear	NN	O	O
targeting	NN	O	O
.	NN	O	O

Binding	NN	O	O
site	NN	O	O
selection	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
alternating	NN	O	B-protein
RCGCRYGCGY	NN	O	I-protein
consensus	NN	O	I-protein
constitutes	NN	O	O
high-affinity	NN	O	O
IBR/F	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
and	NN	O	O
that	NN	O	O
the	NN	O	O
direct-repeat	NN	O	B-DNA
palindrome	NN	O	I-DNA
TGCGCATGCGCA	NN	O	O
is	NN	O	O
the	NN	O	O
optimal	NN	O	B-DNA
site	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
survey	NN	O	O
of	NN	O	O
genes	NN	O	B-DNA
potentially	NN	O	O
regulated	NN	O	O
by	NN	O	O
this	NN	O	O
family	NN	O	O
of	NN	O	O
factors	NN	O	B-protein
primarily	NN	O	O
revealed	NN	O	O
genes	NN	O	B-DNA
involved	NN	O	O
in	NN	O	O
growth-related	NN	O	O
metabolism	NN	O	O
.	NN	O	O

-DOCSTART-	O

Triggering	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	O
interleukin-6	NN	O	O
gene	NN	O	O
by	NN	O	O
interferon-gamma	NN	O	O
and	NN	O	O
tumor	NN	O	O
necrosis	NN	O	O
factor-alpha	NN	O	O
in	NN	O	O
monocytic	NN	O	O
cells	NN	O	O
involves	NN	O	O
cooperation	NN	O	O
between	NN	O	O
interferon	NN	O	B-protein
regulatory	NN	O	I-protein
factor-1	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
and	NN	O	O
Sp1	NN	O	B-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

We	NN	O	O
investigated	NN	O	O
the	NN	O	O
molecular	NN	O	O
basis	NN	O	O
of	NN	O	O
the	NN	O	O
synergistic	NN	O	O
induction	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
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
of	NN	O	O
human	NN	O	B-DNA
interleukin-6	NN	O	I-DNA
(	NN	O	I-DNA
IL-6	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
THP-1	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
and	NN	O	O
compared	NN	O	O
it	NN	O	O
with	NN	O	O
the	NN	O	O
basis	NN	O	O
of	NN	O	O
this	NN	O	O
induction	NN	O	O
by	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
.	NN	O	O

Functional	NN	O	O
studies	NN	O	O
with	NN	O	O
IL-6	NN	O	B-DNA
promoter	NN	O	I-DNA
demonstrated	NN	O	O
that	NN	O	O
three	NN	O	O
regions	NN	O	O
are	NN	O	O
the	NN	O	O
targets	NN	O	O
of	NN	O	O
the	NN	O	O
IFN-gamma	NN	O	B-protein
and/or	NN	O	O
TNF-alpha	NN	O	B-protein
action	NN	O	O
,	NN	O	O
whereas	NN	O	O
only	NN	O	O
one	NN	O	O
of	NN	O	O
these	NN	O	O
regions	NN	O	O
seemed	NN	O	O
to	NN	O	O
be	NN	O	O
implicated	NN	O	O
in	NN	O	O
LPS	NN	O	O
activation	NN	O	O
.	NN	O	O

The	NN	O	O
three	NN	O	O
regions	NN	O	O
concerned	NN	O	O
are	NN	O	O
:	NN	O	O
1	NN	O	O
)	NN	O	O
a	NN	O	O
region	NN	O	O
between	NN	O	O
-73	NN	O	O
and	NN	O	O
-36	NN	O	O
,	NN	O	O
which	NN	O	O
is	NN	O	O
the	NN	O	O
minimal	NN	O	B-DNA
element	NN	O	I-DNA
inducible	NN	O	O
by	NN	O	O
LPS	NN	O	O
or	NN	O	O
TNF-alpha	NN	O	B-protein
;	NN	O	O
2	NN	O	O
)	NN	O	O
an	NN	O	O
element	NN	O	O
located	NN	O	O
between	NN	O	O
-181	NN	O	O
and	NN	O	O
-73	NN	O	O
,	NN	O	O
which	NN	O	O
appeared	NN	O	O
to	NN	O	O
regulate	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
TNF-alpha	NN	O	B-protein
negatively	NN	O	O
;	NN	O	O
and	NN	O	O
3	NN	O	O
)	NN	O	O
a	NN	O	O
distal	NN	O	B-DNA
element	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
-224	NN	O	O
,	NN	O	O
which	NN	O	O
was	NN	O	O
inducible	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
alone	NN	O	O
.	NN	O	O

LPS	NN	O	O
signaling	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
involve	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
activation	NN	O	O
by	NN	O	O
the	NN	O	O
p50/p65	NN	O	B-protein
heterodimers	NN	O	I-protein
.	NN	O	O

Synergistic	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
by	NN	O	O
IFN-gamma	NN	O	B-protein
and	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
involved	NN	O	O
cooperation	NN	O	O
between	NN	O	O
the	NN	O	O
IRF-1	NN	O	B-protein
and	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
p65	NN	O	I-protein
homodimers	NN	O	I-protein
with	NN	O	O
concomitant	NN	O	O
removal	NN	O	O
of	NN	O	O
the	NN	O	O
negative	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
retinoblastoma	NN	O	B-DNA
control	NN	O	I-DNA
element	NN	O	I-DNA
present	NN	O	O
in	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
removal	NN	O	O
occurred	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
constitutive	NN	O	B-protein
Sp1	NN	O	I-protein
factor	NN	O	I-protein
,	NN	O	O
whose	NN	O	O
increased	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
phosphorylation	NN	O	O
were	NN	O	O
mediated	NN	O	O
by	NN	O	O
IFN-gamma	NN	O	B-protein
.	NN	O	O

-DOCSTART-	O

Mutation	NN	O	O
of	NN	O	O
Jak3	NN	O	B-protein
in	NN	O	O
a	NN	O	O
patient	NN	O	O
with	NN	O	O
SCID	NN	O	O
:	NN	O	O
essential	NN	O	O
role	NN	O	O
of	NN	O	O
Jak3	NN	O	B-protein
in	NN	O	O
lymphoid	NN	O	O
development	NN	O	O
.	NN	O	O

Males	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
XSCID	NN	O	O
)	NN	O	O
have	NN	O	O
defects	NN	O	O
in	NN	O	O
the	NN	O	O
common	NN	O	O
cytokine	NN	O	B-DNA
receptor	NN	O	I-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
that	NN	O	O
encodes	NN	O	O
a	NN	O	O
shared	NN	O	O
,	NN	O	O
essential	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
receptors	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
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

The	NN	O	O
Janus	NN	O	B-protein
family	NN	O	I-protein
tyrosine	NN	O	I-protein
kinase	NN	O	I-protein
Jak3	NN	O	B-protein
is	NN	O	O
the	NN	O	O
only	NN	O	O
signaling	NN	O	O
molecule	NN	O	O
known	NN	O	O
to	NN	O	O
be	NN	O	O
associated	NN	O	O
with	NN	O	O
gamma	NN	O	B-protein
c	NN	O	I-protein
,	NN	O	O
so	NN	O	O
it	NN	O	O
was	NN	O	O
hypothesized	NN	O	O
that	NN	O	O
defects	NN	O	O
in	NN	O	O
Jak3	NN	O	B-protein
might	NN	O	O
cause	NN	O	O
an	NN	O	O
XSCID-like	NN	O	O
phenotype	NN	O	O
.	NN	O	O

A	NN	O	O
girl	NN	O	O
with	NN	O	O
immunological	NN	O	O
features	NN	O	O
indistinguishable	NN	O	O
from	NN	O	O
those	NN	O	O
of	NN	O	O
XSCID	NN	O	O
was	NN	O	O
therefore	NN	O	O
selected	NN	O	O
for	NN	O	O
analysis	NN	O	O
.	NN	O	O

An	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
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
derived	NN	O	O
from	NN	O	O
her	NN	O	O
lymphocytes	NN	O	B-cell_type
had	NN	O	O
normal	NN	O	O
gamma	NN	O	B-protein
c	NN	O	I-protein
expression	NN	O	O
but	NN	O	O
lacked	NN	O	O
Jak3	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
had	NN	O	O
greatly	NN	O	O
diminished	NN	O	O
Jak3	NN	O	B-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
.	NN	O	O

Sequencing	NN	O	O
revealed	NN	O	O
a	NN	O	O
different	NN	O	O
mutation	NN	O	O
on	NN	O	O
each	NN	O	O
allele	NN	O	O
:	NN	O	O
a	NN	O	O
single	NN	O	O
nucleotide	NN	O	O
insertion	NN	O	O
resulting	NN	O	O
in	NN	O	O
a	NN	O	O
frame	NN	O	O
shift	NN	O	O
and	NN	O	O
premature	NN	O	O
termination	NN	O	O
in	NN	O	O
the	NN	O	O
Jak3	NN	O	B-protein
JH4	NN	O	I-protein
domain	NN	O	I-protein
and	NN	O	O
a	NN	O	O
nonsense	NN	O	O
mutation	NN	O	O
in	NN	O	O
the	NN	O	O
Jak3	NN	O	B-protein
JH2	NN	O	O
domain	NN	O	O
.	NN	O	O

The	NN	O	O
lack	NN	O	O
of	NN	O	O
Jak3	NN	O	B-protein
expression	NN	O	O
correlated	NN	O	O
with	NN	O	O
impaired	NN	O	O
B	NN	O	O
cell	NN	O	O
signaling	NN	O	O
,	NN	O	O
as	NN	O	O
demonstrated	NN	O	O
by	NN	O	O
the	NN	O	O
inability	NN	O	O
of	NN	O	O
IL-4	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
Stat6	NN	O	B-protein
in	NN	O	O
the	NN	O	O
EBV-transformed	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
from	NN	O	O
the	NN	O	O
patient.	NN	O	O
These	NN	O	O
observations	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
functions	NN	O	O
of	NN	O	O
gamma	NN	O	B-protein
c	NN	O	I-protein
are	NN	O	O
dependent	NN	O	O
on	NN	O	O
Jak3	NN	O	B-protein
and	NN	O	O
that	NN	O	O
Jak3	NN	O	B-protein
is	NN	O	O
essential	NN	O	O
for	NN	O	O
lymphoid	NN	O	O
development	NN	O	O
and	NN	O	O
signaling	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
overexpression	NN	O	O
of	NN	O	O
the	NN	O	O
L-selectin	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
fresh	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
adult	NN	O	O
T-cell	NN	O	O
leukemia	NN	O	O
that	NN	O	O
can	NN	O	O
be	NN	O	O
transactivated	NN	O	O
by	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
.	NN	O	O

L-selectin	NN	O	B-protein
is	NN	O	O
an	NN	O	O
adhesion	NN	O	O
molecule	NN	O	O
of	NN	O	O
the	NN	O	O
selectin	NN	O	O
family	NN	O	O
that	NN	O	O
mediates	NN	O	O
the	NN	O	O
initial	NN	O	O
step	NN	O	O
of	NN	O	O
leukocyte	NN	O	O
adhesion	NN	O	O
to	NN	O	O
vascular	NN	O	O
endothelium	NN	O	O
.	NN	O	O

Upon	NN	O	O
cellular	NN	O	O
activation	NN	O	O
,	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
L-selectin	NN	O	B-DNA
gene	NN	O	I-DNA
is	NN	O	O
downregulated	NN	O	O
at	NN	O	O
both	NN	O	O
the	NN	O	O
protein	NN	O	O
and	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

To	NN	O	O
understand	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
leukemic	NN	O	B-cell_type
cell	NN	O	I-cell_type
infiltration	NN	O	O
into	NN	O	O
organs	NN	O	O
,	NN	O	O
we	NN	O	O
studied	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
L-selectin	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
fresh	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
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
patients	NN	O	O
and	NN	O	O
investigated	NN	O	O
the	NN	O	O
response	NN	O	O
of	NN	O	O
the	NN	O	O
L-selectin	NN	O	B-DNA
promoter	NN	O	I-DNA
to	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
(	NN	O	I-protein
HTLV-1	NN	O	I-protein
)	NN	O	I-protein
Tax	NN	O	I-protein
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
viral	NN	O	B-protein
transcriptional	NN	O	I-protein
transactivator	NN	O	I-protein
.	NN	O	O

Flow	NN	O	O
cytometry	NN	O	O
showed	NN	O	O
that	NN	O	O
L-selectin	NN	O	B-protein
was	NN	O	O
expressed	NN	O	O
on	NN	O	O
fresh	NN	O	O
ATL	NN	O	B-cell_line
cells	NN	O	I-cell_line
along	NN	O	O
with	NN	O	O
other	NN	O	O
activation	NN	O	B-protein
antigens	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
ATL	NN	O	B-cell_line
cells	NN	O	I-cell_line
overexpressed	NN	O	O
that	NN	O	O
L-selectin	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
that	NN	O	O
the	NN	O	O
level	NN	O	O
was	NN	O	O
aberrantly	NN	O	O
upregulated	NN	O	O
after	NN	O	O
PMA	NN	O	O
stimulation	NN	O	O
.	NN	O	O

Studies	NN	O	O
using	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
showed	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
L-selectin	NN	O	B-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
the	NN	O	O
infiltrating	NN	O	O
leukemic	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
liver	NN	O	O
of	NN	O	O
two	NN	O	O
ATL	NN	O	O
patients	NN	O	O
.	NN	O	O

Intravenous	NN	O	O
injection	NN	O	O
of	NN	O	O
a	NN	O	O
rat	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
line	NN	O	I-cell_line
that	NN	O	O
overexpresses	NN	O	O
L-selectin	NN	O	B-protein
showed	NN	O	O
increased	NN	O	O
organ	NN	O	O
infiltration	NN	O	O
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
Tax	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
JPX9	NN	O	B-cell_line
cells	NN	O	I-cell_line
resulted	NN	O	O
in	NN	O	O
about	NN	O	O
a	NN	O	O
twofold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
expression	NN	O	O
levels	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
basal	NN	O	O
level	NN	O	O
.	NN	O	O

Chloramphenicol	NN	O	B-protein
acetyltransferase	NN	O	I-protein
(	NN	O	O
CAT	NN	O	B-protein
)	NN	O	O
assay	NN	O	O
after	NN	O	O
transient	NN	O	O
cotransfection	NN	O	O
showed	NN	O	O
about	NN	O	O
a	NN	O	O
fivefold	NN	O	O
transactivation	NN	O	O
of	NN	O	O
the	NN	O	O
L-selectin	NN	O	B-DNA
promoter	NN	O	I-DNA
by	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

The	NN	O	O
serum	NN	O	O
level	NN	O	O
of	NN	O	O
the	NN	O	O
shed	NN	O	O
form	NN	O	O
of	NN	O	O
L-selectin	NN	O	B-protein
was	NN	O	O
significantly	NN	O	O
increased	NN	O	O
in	NN	O	O
ATL	NN	O	O
patients	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SD	NN	O	O
,	NN	O	O
4	NN	O	O
,	NN	O	O
215.4	NN	O	O
+/-	NN	O	O
4	NN	O	O
,	NN	O	O
111	NN	O	O
ng/mL	NN	O	O
)	NN	O	O
compared	NN	O	O
with	NN	O	O
those	NN	O	O
of	NN	O	O
asymptomatic	NN	O	O
carriers	NN	O	O
and	NN	O	O
healthy	NN	O	O
blood	NN	O	O
donors	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SD	NN	O	O
,	NN	O	O
1	NN	O	O
,	NN	O	O
148.0	NN	O	O
+/-	NN	O	O
269.0	NN	O	O
ng/mL	NN	O	O
and	NN	O	O
991.9	NN	O	O
+/-	NN	O	O
224	NN	O	O
ng/mL	NN	O	O
,	NN	O	O
respectively	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
ATL	NN	O	B-cell_line
cells	NN	O	I-cell_line
constitutively	NN	O	O
overexpress	NN	O	O
the	NN	O	O
L-selectin	NN	O	B-DNA
gene	NN	O	I-DNA
that	NN	O	O
can	NN	O	O
be	NN	O	O
transactivated	NN	O	O
by	NN	O	O
HTLV-1	NN	O	O
Tax	NN	O	B-protein
.	NN	O	O

The	NN	O	O
overexpression	NN	O	O
of	NN	O	O
L-selectin	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
of	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
,	NN	O	O
by	NN	O	O
ATL	NN	O	B-cell_line
cells	NN	O	I-cell_line
may	NN	O	O
provide	NN	O	O
a	NN	O	O
basis	NN	O	O
for	NN	O	O
ATL	NN	O	B-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
attach	NN	O	O
the	NN	O	O
vascular	NN	O	O
endothelium	NN	O	O
,	NN	O	O
leading	NN	O	O
to	NN	O	O
transmigration	NN	O	O
and	NN	O	O
organ	NN	O	O
infitration	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
herpesvirus	NN	O	O
6	NN	O	O
variant	NN	O	O
A	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
variant	NN	O	O
B	NN	O	O
,	NN	O	O
infects	NN	O	O
EBV-positive	NN	O	B-cell_line
B	NN	O	I-cell_line
lymphoid	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
activating	NN	O	O
the	NN	O	O
latent	NN	O	B-DNA
EBV	NN	O	I-DNA
genome	NN	O	I-DNA
through	NN	O	O
a	NN	O	O
BZLF-1	NN	O	B-protein
-dependent	NN	O	O
mechanism	NN	O	O
.	NN	O	O

Human	NN	O	O
herpesvirus	NN	O	O
6	NN	O	O
,	NN	O	O
a	NN	O	O
predominantly	NN	O	O
T	NN	O	O
lymphotropic	NN	O	O
virus	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
recently	NN	O	O
shown	NN	O	O
to	NN	O	O
infect	NN	O	O
some	NN	O	O
EBV-positive	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
and	NN	O	O
to	NN	O	O
induce	NN	O	O
in	NN	O	O
them	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	O
lytic	NN	O	O
cycle	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
have	NN	O	O
confirmed	NN	O	O
and	NN	O	O
extended	NN	O	O
such	NN	O	O
observations	NN	O	O
,	NN	O	O
showing	NN	O	O
that	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
this	NN	O	O
phenomenon	NN	O	O
is	NN	O	O
restricted	NN	O	O
to	NN	O	O
the	NN	O	O
variant	NN	O	O
A	NN	O	O
of	NN	O	O
HHV-6	NN	O	O
:	NN	O	O
in	NN	O	O
fact	NN	O	O
two	NN	O	O
isolates	NN	O	O
belonging	NN	O	O
to	NN	O	O
the	NN	O	O
HHV-6	NN	O	O
variant	NN	O	O
B	NN	O	O
(	NN	O	O
BA92	NN	O	O
and	NN	O	O
Z29	NN	O	O
)	NN	O	O
were	NN	O	O
neither	NN	O	O
able	NN	O	O
to	NN	O	O
infect	NN	O	O
any	NN	O	O
B	NN	O	B-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
,	NN	O	O
independently	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	O
status	NN	O	O
,	NN	O	O
nor	NN	O	O
to	NN	O	O
induce	NN	O	O
the	NN	O	O
EBV	NN	O	O
genome	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
only	NN	O	O
exception	NN	O	O
is	NN	O	O
represented	NN	O	O
by	NN	O	O
the	NN	O	O
P3HR1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
in	NN	O	O
which	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
infection	NN	O	O
by	NN	O	O
the	NN	O	O
variant	NN	O	O
B	NN	O	O
does	NN	O	O
not	NN	O	O
determine	NN	O	O
induction	NN	O	O
of	NN	O	O
EBV	NN	O	B-protein
antigens	NN	O	I-protein
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
EBV	NN	O	B-DNA
genome	NN	O	I-DNA
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
susceptibility	NN	O	O
of	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
to	NN	O	O
HHV-6	NN	O	O
infection	NN	O	O
,	NN	O	O
increasing	NN	O	O
the	NN	O	O
binding	NN	O	O
sites	NN	O	O
and	NN	O	O
the	NN	O	O
percentage	NN	O	O
of	NN	O	O
infectable	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
as	NN	O	O
detected	NN	O	O
by	NN	O	O
immunoelectron	NN	O	O
microscopy	NN	O	O
;	NN	O	O
and	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
HHV-6	NN	O	B-cell_line
infected	NN	O	I-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
transfected	NN	O	O
with	NN	O	O
plasmids	NN	O	B-DNA
bearing	NN	O	O
the	NN	O	O
promoter	NN	O	B-DNA
regions	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
EBV	NN	O	O
early	NN	O	O
genes	NN	O	O
BZLF1	NN	O	O
and	NN	O	O
BMRF1	NN	O	B-DNA
,	NN	O	O
show	NN	O	O
a	NN	O	O
strong	NN	O	O
transactivation	NN	O	O
of	NN	O	O
these	NN	O	O
promoters	NN	O	B-DNA
.	NN	O	O

-DOCSTART-	O

Evidence	NN	O	O
for	NN	O	O
normal	NN	O	O
vitamin	NN	O	B-RNA
D	NN	O	I-RNA
receptor	NN	O	I-RNA
messenger	NN	O	I-RNA
ribonucleic	NN	O	I-RNA
acid	NN	O	I-RNA
and	NN	O	O
genotype	NN	O	O
in	NN	O	O
absorptive	NN	O	O
hypercalciuria	NN	O	O
.	NN	O	O

Absorptive	NN	O	O
hypercalciuria	NN	O	O
(	NN	O	O
a	NN	O	O
stone-forming	NN	O	O
condition	NN	O	O
)	NN	O	O
is	NN	O	O
characterized	NN	O	O
by	NN	O	O
gut	NN	O	O
hyperabsorption	NN	O	O
of	NN	O	O
calcium	NN	O	O
,	NN	O	O
hypercalciuria	NN	O	O
,	NN	O	O
and	NN	O	O
reduced	NN	O	O
bone	NN	O	O
density	NN	O	O
.	NN	O	O

Inasmuch	NN	O	O
as	NN	O	O
these	NN	O	O
features	NN	O	O
implicate	NN	O	O
enhanced	NN	O	O
calcitriol	NN	O	O
action	NN	O	O
in	NN	O	O
gut	NN	O	O
and	NN	O	O
bone	NN	O	O
,	NN	O	O
we	NN	O	O
analyzed	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
to	NN	O	O
ascertain	NN	O	O
whether	NN	O	O
an	NN	O	O
abnormality	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
marks	NN	O	O
patients	NN	O	O
with	NN	O	O
intestinal	NN	O	O
hyperabsorption	NN	O	O
of	NN	O	O
calcium	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
compared	NN	O	O
the	NN	O	O
frequency	NN	O	O
of	NN	O	O
a	NN	O	O
restriction	NN	O	B-DNA
fragment	NN	O	I-DNA
length	NN	O	I-DNA
polymorphism	NN	O	I-DNA
(	NN	O	O
Bsm	NN	O	O
I	NN	O	O
)	NN	O	O
associated	NN	O	O
with	NN	O	O
different	NN	O	O
alleles	NN	O	O
of	NN	O	O
the	NN	O	O
VDR	NN	O	B-DNA
gene	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
33	NN	O	O
well	NN	O	O
characterized	NN	O	O
absorptive	NN	O	O
hypercalciuric	NN	O	O
patients	NN	O	O
and	NN	O	O
a	NN	O	O
group	NN	O	O
of	NN	O	O
36	NN	O	O
normal	NN	O	O
race-	NN	O	O
and	NN	O	O
age-matched	NN	O	O
control	NN	O	O
subjects	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
no	NN	O	O
difference	NN	O	O
between	NN	O	O
the	NN	O	O
distribution	NN	O	O
of	NN	O	O
the	NN	O	O
VDR	NN	O	B-protein
alleles	NN	O	O
in	NN	O	O
the	NN	O	O
patient	NN	O	O
population	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
normal	NN	O	O
population	NN	O	O
.	NN	O	O

The	NN	O	O
coding	NN	O	O
region	NN	O	O
of	NN	O	O
VDR	NN	O	B-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
was	NN	O	O
also	NN	O	O
normal	NN	O	O
,	NN	O	O
as	NN	O	O
determined	NN	O	O
by	NN	O	O
both	NN	O	O
DNA	NN	O	B-DNA
sequence	NN	O	I-DNA
analysis	NN	O	O
and	NN	O	O
chemical	NN	O	O
mismatch	NN	O	O
cleavage	NN	O	O
analysis	NN	O	O
of	NN	O	O
copy	NN	O	O
DNA	NN	O	O
from	NN	O	O
11	NN	O	O
index	NN	O	O
absorptive	NN	O	O
hypercalciuric	NN	O	O
patients	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
results	NN	O	O
,	NN	O	O
we	NN	O	O
propose	NN	O	O
that	NN	O	O
the	NN	O	O
enhanced	NN	O	O
intestinal	NN	O	O
calcium	NN	O	O
absorption	NN	O	O
invariably	NN	O	O
seen	NN	O	O
in	NN	O	O
absorptive	NN	O	O
hypercalciuria	NN	O	O
and	NN	O	O
attendant	NN	O	O
symptoms	NN	O	O
of	NN	O	O
this	NN	O	O
disorder	NN	O	O
are	NN	O	O
not	NN	O	O
attributable	NN	O	O
to	NN	O	O
mutations	NN	O	O
of	NN	O	O
the	NN	O	O
VDR	NN	O	B-protein
and	NN	O	O
are	NN	O	O
not	NN	O	O
linked	NN	O	O
to	NN	O	O
a	NN	O	O
common	NN	O	O
VDR	NN	O	B-DNA
genotype	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
gene	NN	O	O
encoding	NN	O	O
the	NN	O	O
human	NN	O	B-protein
C-type	NN	O	I-protein
lectin	NN	O	I-protein
leukocyte	NN	O	I-protein
receptor	NN	O	I-protein
AIM/CD69	NN	O	B-protein
and	NN	O	O
functional	NN	O	O
characterization	NN	O	O
of	NN	O	O
its	NN	O	O
tumor	NN	O	B-DNA
necrosis	NN	O	I-DNA
factor-alpha-responsive	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
human	NN	O	B-protein
activation	NN	O	I-protein
antigen	NN	O	I-protein
CD69	NN	O	B-protein
is	NN	O	O
a	NN	O	O
member	NN	O	O
of	NN	O	O
the	NN	O	O
C-type	NN	O	B-protein
animal	NN	O	I-protein
lectin	NN	O	I-protein
superfamily	NN	O	I-protein
that	NN	O	O
functions	NN	O	O
as	NN	O	O
a	NN	O	O
signal-transmitting	NN	O	B-protein
receptor	NN	O	I-protein
.	NN	O	O

Although	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CD69	NN	O	B-protein
can	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
vitro	NN	O	O
on	NN	O	O
cells	NN	O	O
of	NN	O	O
most	NN	O	O
hematopoietic	NN	O	O
lineages	NN	O	O
with	NN	O	O
a	NN	O	O
wide	NN	O	O
variety	NN	O	O
of	NN	O	O
stimuli	NN	O	O
,	NN	O	O
in	NN	O	O
vivo	NN	O	O
it	NN	O	O
is	NN	O	O
mainly	NN	O	O
expressed	NN	O	O
by	NN	O	O
T-lymphocytes	NN	O	B-cell_type
located	NN	O	O
in	NN	O	O
the	NN	O	O
inflammatory	NN	O	O
infiltrates	NN	O	O
of	NN	O	O
several	NN	O	O
human	NN	O	O
diseases	NN	O	O
.	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
constitutive	NN	O	O
and	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
CD69	NN	O	B-protein
by	NN	O	O
leukocytes	NN	O	B-cell_type
,	NN	O	O
we	NN	O	O
isolated	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
CD69	NN	O	B-protein
gene	NN	O	O
and	NN	O	O
carried	NN	O	O
out	NN	O	O
its	NN	O	O
functional	NN	O	O
characterization	NN	O	O
.	NN	O	O

Sequence	NN	O	O
analysis	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
CD69	NN	O	B-DNA
gene	NN	O	I-DNA
revealed	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
potential	NN	O	O
TATA	NN	O	B-DNA
element	NN	O	I-DNA
30	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	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
and	NN	O	O
several	NN	O	O
putative	NN	O	O
binding	NN	O	O
sequences	NN	O	O
for	NN	O	O
inducible	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
(	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
Egr-1	NN	O	B-protein
,	NN	O	O
AP-1	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
might	NN	O	O
mediate	NN	O	O
the	NN	O	O
inducible	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
.	NN	O	O

Transient	NN	O	O
expression	NN	O	O
of	NN	O	O
CD69	NN	O	B-DNA
promoter-based	NN	O	I-DNA
reporter	NN	O	I-DNA
gene	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
indicated	NN	O	O
that	NN	O	O
the	NN	O	O
proximal	NN	O	B-DNA
promoter	NN	O	I-DNA
region	NN	O	I-DNA
spanning	NN	O	O
positions	NN	O	O
-78	NN	O	O
to	NN	O	O
+16	NN	O	O
contained	NN	O	O
the	NN	O	O
cis-acting	NN	O	B-DNA
sequences	NN	O	I-DNA
necessary	NN	O	O
for	NN	O	O
basal	NN	O	O
and	NN	O	O
phorbol	NN	O	O
12-myristate	NN	O	O
13-acetate-inducible	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
CD69	NN	O	B-DNA
gene	NN	O	I-DNA
.	NN	O	O

Removal	NN	O	O
of	NN	O	O
the	NN	O	O
upstream	NN	O	O
sequences	NN	O	O
located	NN	O	O
between	NN	O	O
positions	NN	O	O
-78	NN	O	O
and	NN	O	O
-38	NN	O	O
resulted	NN	O	O
in	NN	O	O
decreased	NN	O	O
promoter	NN	O	O
strength	NN	O	O
and	NN	O	O
abolished	NN	O	O
the	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

We	NN	O	O
also	NN	O	O
found	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
is	NN	O	O
capable	NN	O	O
of	NN	O	O
inducing	NN	O	O
the	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
CD69	NN	O	B-protein
molecule	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
promoter	NN	O	O
activity	NN	O	O
of	NN	O	O
fusion	NN	O	O
plasmids	NN	O	O
that	NN	O	O
contain	NN	O	O
5'-flanking	NN	O	O
sequences	NN	O	O
of	NN	O	O
the	NN	O	O
CD69	NN	O	B-protein
gene	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
cytokine	NN	O	O
may	NN	O	O
regulate	NN	O	O
in	NN	O	O
vivo	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CD69	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

Characterization	NN	O	O
of	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
end	NN	O	I-DNA
of	NN	O	I-DNA
human	NN	O	I-DNA
thromboxane	NN	O	I-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Organizational	NN	O	O
analysis	NN	O	O
and	NN	O	O
mapping	NN	O	O
of	NN	O	O
protein	NN	O	B-DNA
kinase	NN	O	I-DNA
C	NN	O	I-DNA
--	NN	O	I-DNA
responsive	NN	O	I-DNA
elements	NN	O	I-DNA
regulating	NN	O	O
expression	NN	O	O
in	NN	O	O
platelets	NN	O	B-cell_type
.	NN	O	O

Platelet	NN	O	B-protein
thromboxane	NN	O	I-protein
receptors	NN	O	I-protein
are	NN	O	O
acutely	NN	O	O
and	NN	O	O
reversibly	NN	O	O
upregulated	NN	O	O
after	NN	O	O
acute	NN	O	O
myocardial	NN	O	O
infarction	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
if	NN	O	O
platelet	NN	O	O
thromboxane	NN	O	B-protein
receptors	NN	O	I-protein
are	NN	O	O
under	NN	O	O
transcriptional	NN	O	O
control	NN	O	O
,	NN	O	O
we	NN	O	O
isolated	NN	O	O
and	NN	O	O
characterized	NN	O	O
human	NN	O	B-DNA
genomic	NN	O	I-DNA
DNA	NN	O	I-DNA
clones	NN	O	I-DNA
containing	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
thromboxane	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
exon-intron	NN	O	B-DNA
structure	NN	O	I-DNA
of	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
the	NN	O	O
thromboxane	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
determined	NN	O	O
initially	NN	O	O
by	NN	O	O
comparing	NN	O	O
the	NN	O	O
nucleotide	NN	O	B-DNA
sequence	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
genomic	NN	O	I-DNA
clone	NN	O	I-DNA
with	NN	O	O
that	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	B-DNA
human	NN	O	I-DNA
uterine	NN	O	I-DNA
thromboxane	NN	O	I-DNA
receptor	NN	O	I-DNA
cDNA	NN	O	I-DNA
that	NN	O	O
extended	NN	O	O
the	NN	O	O
mRNA	NN	O	B-RNA
141	NN	O	B-DNA
bp	NN	O	I-DNA
further	NN	O	I-DNA
upstream	NN	O	I-DNA
than	NN	O	O
the	NN	O	O
previously	NN	O	B-DNA
identified	NN	O	I-DNA
human	NN	O	I-DNA
placental	NN	O	I-DNA
cDNA	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
major	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
was	NN	O	O
located	NN	O	O
in	NN	O	O
three	NN	O	O
human	NN	O	O
tissues	NN	O	O
approximately	NN	O	O
560	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
initiation	NN	O	I-DNA
codon	NN	O	I-DNA
and	NN	O	O
380	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
from	NN	O	O
any	NN	O	O
previously	NN	O	O
identified	NN	O	O
transcription	NN	O	B-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
thromboxane	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
has	NN	O	O
neither	NN	O	O
a	NN	O	O
TATA	NN	O	B-DNA
nor	NN	O	O
a	NN	O	O
CAAT	NN	O	B-DNA
consensus	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Promoter	NN	O	O
function	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
the	NN	O	O
thromboxane	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
was	NN	O	O
evaluated	NN	O	O
by	NN	O	O
transfection	NN	O	O
of	NN	O	O
thromboxane	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
promoter/chloramphenicol	NN	O	I-DNA
acetyltransferase	NN	O	I-DNA
(	NN	O	I-DNA
CAT	NN	O	I-DNA
)	NN	O	I-DNA
chimera	NN	O	I-DNA
plasmids	NN	O	I-DNA
into	NN	O	O
platelet-like	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Thromboxane	NN	O	O
receptor	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
,	NN	O	O
as	NN	O	O
assessed	NN	O	O
by	NN	O	O
CAT	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
was	NN	O	O
relatively	NN	O	O
weak	NN	O	O
but	NN	O	O
was	NN	O	O
significantly	NN	O	O
enhanced	NN	O	O
by	NN	O	O
phorbol	NN	O	O
ester	NN	O	O
treatment	NN	O	O
.	NN	O	O

Functional	NN	O	O
analysis	NN	O	O
of	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
deletion	NN	O	I-DNA
constructs	NN	O	I-DNA
in	NN	O	O
transfected	NN	O	O
K562	NN	O	B-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
gel	NN	O	O
mobility	NN	O	O
shift	NN	O	O
localized	NN	O	O
the	NN	O	O
major	NN	O	O
phorbol	NN	O	O
ester-responsive	NN	O	O
motifs	NN	O	O
in	NN	O	O
the	NN	O	O
thromboxane	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	O
to	NN	O	O
a	NN	O	O
cluster	NN	O	O
of	NN	O	O
activator	NN	O	B-DNA
protein-2	NN	O	I-DNA
(	NN	O	I-DNA
AP-2	NN	O	I-DNA
)	NN	O	I-DNA
binding	NN	O	I-DNA
consensus	NN	O	I-DNA
sites	NN	O	I-DNA
located	NN	O	O
approximately	NN	O	O
1.8	NN	O	O
kb	NN	O	O
5	NN	O	O
'	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

These	NN	O	O
studies	NN	O	O
are	NN	O	O
the	NN	O	O
first	NN	O	O
to	NN	O	O
determine	NN	O	O
the	NN	O	O
structure	NN	O	O
and	NN	O	O
organization	NN	O	O
of	NN	O	O
the	NN	O	O
5	NN	O	O
'	NN	O	O
end	NN	O	O
of	NN	O	O
the	NN	O	O
thromboxane	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
and	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
thromboxane	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
expression	NN	O	O
can	NN	O	O
be	NN	O	O
regulated	NN	O	O
by	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
via	NN	O	O
induction	NN	O	O
of	NN	O	O
an	NN	O	O
AP-2-like	NN	O	B-protein
nuclear	NN	O	I-protein
factor	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
upstream	NN	O	B-DNA
promoter	NN	O	I-DNA
elements	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
findings	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
mechanism	NN	O	O
for	NN	O	O
previously	NN	O	O
described	NN	O	O
upregulation	NN	O	O
of	NN	O	O
platelet	NN	O	B-protein
thromboxane	NN	O	I-protein
receptors	NN	O	I-protein
after	NN	O	O
acute	NN	O	O
myocardial	NN	O	O
infarction	NN	O	O
is	NN	O	O
increased	NN	O	O
thromboxane	NN	O	B-DNA
receptor	NN	O	I-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
platelet-progenitor	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Estrogen	NN	O	B-protein
receptor	NN	O	I-protein
concentration	NN	O	O
and	NN	O	O
social	NN	O	O
factors	NN	O	O
as	NN	O	O
predictors	NN	O	O
of	NN	O	O
natural	NN	O	O
killer	NN	O	O
cell	NN	O	O
activity	NN	O	O
in	NN	O	O
early-stage	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
.	NN	O	O

Confirmation	NN	O	O
of	NN	O	O
a	NN	O	O
model	NN	O	O
.	NN	O	O

Previous	NN	O	O
work	NN	O	O
of	NN	O	O
ours	NN	O	O
has	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
a	NN	O	O
significant	NN	O	O
amount	NN	O	O
of	NN	O	O
natural	NN	O	O
killer	NN	O	O
(	NN	O	O
NK	NN	O	O
)	NN	O	O
activity	NN	O	O
variance	NN	O	O
after	NN	O	O
surgery	NN	O	O
in	NN	O	O
stage	NN	O	O
I	NN	O	O
and	NN	O	O
II	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
could	NN	O	O
be	NN	O	O
accounted	NN	O	O
for	NN	O	O
by	NN	O	O
both	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
status	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	O
and	NN	O	O
by	NN	O	O
social	NN	O	O
factors	NN	O	O
,	NN	O	O
namely	NN	O	O
,	NN	O	O
perceived	NN	O	O
social	NN	O	O
support	NN	O	O
and	NN	O	O
seeking	NN	O	O
social	NN	O	O
support	NN	O	O
as	NN	O	O
a	NN	O	O
general	NN	O	O
coping	NN	O	O
strategy	NN	O	O
.	NN	O	O

As	NN	O	O
considerable	NN	O	O
evidence	NN	O	O
has	NN	O	O
accumulated	NN	O	O
that	NN	O	O
social	NN	O	O
support	NN	O	O
in	NN	O	O
both	NN	O	O
animal	NN	O	O
and	NN	O	O
human	NN	O	O
populations	NN	O	O
may	NN	O	O
have	NN	O	O
survival	NN	O	O
value	NN	O	O
,	NN	O	O
we	NN	O	O
sought	NN	O	O
to	NN	O	O
test	NN	O	O
the	NN	O	O
reliability	NN	O	O
of	NN	O	O
this	NN	O	O
regression	NN	O	O
model	NN	O	O
,	NN	O	O
using	NN	O	O
coping	NN	O	O
and	NN	O	O
perceived	NN	O	O
support	NN	O	O
factor	NN	O	O
values	NN	O	O
obtained	NN	O	O
at	NN	O	O
3	NN	O	O
months	NN	O	O
after	NN	O	O
surgery	NN	O	O
to	NN	O	O
account	NN	O	O
for	NN	O	O
concurrent	NN	O	O
follow-up	NN	O	O
NK	NN	O	O
activity	NN	O	O
in	NN	O	O
this	NN	O	O
serially	NN	O	O
assessed	NN	O	O
group	NN	O	O
of	NN	O	O
patients	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
most	NN	O	O
significant	NN	O	O
variable	NN	O	O
predicting	NN	O	O
NK	NN	O	O
activity	NN	O	O
at	NN	O	O
follow-up	NN	O	O
was	NN	O	O
tumor	NN	O	O
ER	NN	O	B-protein
concentration	NN	O	O
,	NN	O	O
with	NN	O	O
higher	NN	O	O
NK	NN	O	O
activity	NN	O	O
associated	NN	O	O
with	NN	O	O
ER	NN	O	B-protein
-status	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
seeking	NN	O	O
social	NN	O	O
support	NN	O	O
as	NN	O	O
a	NN	O	O
coping	NN	O	O
strategy	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
perceived	NN	O	O
quality	NN	O	O
of	NN	O	O
support	NN	O	O
,	NN	O	O
also	NN	O	O
entered	NN	O	O
the	NN	O	O
model	NN	O	O
to	NN	O	O
account	NN	O	O
for	NN	O	O
a	NN	O	O
significant	NN	O	O
amount	NN	O	O
of	NN	O	O
NK	NN	O	O
activity	NN	O	O
variance	NN	O	O
(	NN	O	O
multivariate	NN	O	O
F	NN	O	O
=	NN	O	O
5.25	NN	O	O
,	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

If	NN	O	O
,	NN	O	O
as	NN	O	O
the	NN	O	O
literature	NN	O	O
suggests	NN	O	O
,	NN	O	O
NK	NN	O	O
activity	NN	O	O
is	NN	O	O
relevant	NN	O	O
to	NN	O	O
breast	NN	O	O
cancer	NN	O	O
control	NN	O	O
,	NN	O	O
and	NN	O	O
since	NN	O	O
ER	NN	O	B-protein
-tumors	NN	O	O
have	NN	O	O
a	NN	O	O
worse	NN	O	O
prognosis	NN	O	O
,	NN	O	O
we	NN	O	O
suggest	NN	O	O
here	NN	O	O
that	NN	O	O
perhaps	NN	O	O
such	NN	O	O
tumors	NN	O	O
are	NN	O	O
resistant	NN	O	O
to	NN	O	O
control	NN	O	O
by	NN	O	O
NK	NN	O	B-cell_type
cells	NN	O	I-cell_type
because	NN	O	O
they	NN	O	O
lack	NN	O	O
the	NN	O	O
ability	NN	O	O
to	NN	O	O
attract	NN	O	O
an	NN	O	O
accumulation	NN	O	O
of	NN	O	O
effector	NN	O	B-cell_type
cells	NN	O	I-cell_type
to	NN	O	O
the	NN	O	O
tumor	NN	O	O
site	NN	O	O
,	NN	O	O
or	NN	O	O
because	NN	O	O
blocking	NN	O	O
factors	NN	O	O
at	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
the	NN	O	O
tumor	NN	O	O
prevent	NN	O	O
local	NN	O	O
tumor	NN	O	O
control	NN	O	O
at	NN	O	O
the	NN	O	O
site	NN	O	O
of	NN	O	O
action	NN	O	O
.	NN	O	O

The	NN	O	O
finding	NN	O	O
related	NN	O	O
to	NN	O	O
social	NN	O	O
support	NN	O	O
also	NN	O	O
replicates	NN	O	O
results	NN	O	O
from	NN	O	O
an	NN	O	O
independent	NN	O	O
sample	NN	O	O
of	NN	O	O
breast	NN	O	O
cancer	NN	O	O
patients	NN	O	O
.	NN	O	O

This	NN	O	O
finding	NN	O	O
,	NN	O	O
taken	NN	O	O
together	NN	O	O
with	NN	O	O
other	NN	O	O
evidence	NN	O	O
that	NN	O	O
this	NN	O	O
social	NN	O	O
variable	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
longer	NN	O	O
survival	NN	O	O
in	NN	O	O
breast	NN	O	O
cancer	NN	O	O
populations	NN	O	O
,	NN	O	O
underscores	NN	O	O
the	NN	O	O
potential	NN	O	O
importance	NN	O	O
of	NN	O	O
this	NN	O	O
social	NN	O	O
support	NN	O	O
variable	NN	O	O
.	NN	O	O

Our	NN	O	O
findings	NN	O	O
also	NN	O	O
suggest	NN	O	O
one	NN	O	O
possible	NN	O	O
immunological	NN	O	O
variable	NN	O	O
involved	NN	O	O
,	NN	O	O
with	NN	O	O
potential	NN	O	O
clinical	NN	O	O
significance	NN	O	O
,	NN	O	O
for	NN	O	O
this	NN	O	O
patient	NN	O	O
population	NN	O	O
.	NN	O	O

-DOCSTART-	O

Solution	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
sequence-specific	NN	O	B-DNA
HMG	NN	O	I-DNA
box	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
lymphocyte	NN	O	B-protein
transcriptional	NN	O	I-protein
activator	NN	O	I-protein
Sox-4	NN	O	B-protein
.	NN	O	O

Two	NN	O	O
groups	NN	O	O
of	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
proteins	NN	O	I-protein
are	NN	O	O
distinguished	NN	O	O
.	NN	O	O

Proteins	NN	O	O
in	NN	O	O
the	NN	O	O
first	NN	O	O
group	NN	O	O
contain	NN	O	O
multiple	NN	O	O
HMG	NN	O	B-protein
boxes	NN	O	I-protein
,	NN	O	O
are	NN	O	O
non-sequence-specific	NN	O	O
,	NN	O	O
and	NN	O	O
recognize	NN	O	O
structural	NN	O	O
features	NN	O	O
as	NN	O	O
found	NN	O	O
in	NN	O	O
cruciform	NN	O	B-DNA
DNA	NN	O	I-DNA
and	NN	O	O
cross-over	NN	O	B-DNA
DNA	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
abundant	NN	O	O
chromosomal	NN	O	B-protein
protein	NN	O	I-protein
HMG-1	NN	O	B-protein
belongs	NN	O	O
to	NN	O	O
this	NN	O	O
subgroup	NN	O	O
.	NN	O	O

Proteins	NN	O	O
in	NN	O	O
the	NN	O	O
second	NN	O	O
group	NN	O	O
carry	NN	O	O
a	NN	O	O
single	NN	O	B-protein
HMG	NN	O	I-protein
box	NN	O	I-protein
with	NN	O	O
affinity	NN	O	O
for	NN	O	O
the	NN	O	O
minor	NN	O	O
groove	NN	O	O
of	NN	O	O
the	NN	O	O
heptamer	NN	O	B-DNA
motif	NN	O	I-DNA
AACAAAG	NN	O	O
or	NN	O	O
variations	NN	O	O
thereof	NN	O	O
.	NN	O	O

A	NN	O	O
solution	NN	O	O
structure	NN	O	O
for	NN	O	O
the	NN	O	O
non-sequence-specific	NN	O	O
C-terminal	NN	O	B-protein
HMG	NN	O	I-protein
box	NN	O	I-protein
of	NN	O	O
HMG-1	NN	O	B-protein
has	NN	O	O
recently	NN	O	O
been	NN	O	O
proposed	NN	O	O
.	NN	O	O

Now	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
the	NN	O	O
solution	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
sequence-specific	NN	O	B-protein
HMG-box	NN	O	I-protein
of	NN	O	O
the	NN	O	O
SRY-related	NN	O	B-protein
protein	NN	O	I-protein
Sox-4	NN	O	B-protein
.	NN	O	O

NMR	NN	O	O
analysis	NN	O	O
demonstrated	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
Val10-Gln22	NN	O	B-protein
,	NN	O	O
Glu30-Leu41	NN	O	B-protein
and	NN	O	O
Phe50-Tyr65	NN	O	B-protein
)	NN	O	O
connected	NN	O	O
by	NN	O	O
loop	NN	O	B-protein
regions	NN	O	I-protein
(	NN	O	O
Ser23-Ala49	NN	O	B-protein
and	NN	O	O
Leu42-Pro49	NN	O	B-protein
)	NN	O	O
.	NN	O	O

Helices	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
II	NN	O	I-protein
are	NN	O	O
positioned	NN	O	O
in	NN	O	O
an	NN	O	O
antiparallel	NN	O	B-protein
mode	NN	O	I-protein
and	NN	O	O
form	NN	O	O
one	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
.	NN	O	O

Helix	NN	O	B-protein
III	NN	O	I-protein
is	NN	O	O
less	NN	O	O
rigid	NN	O	O
,	NN	O	O
makes	NN	O	O
an	NN	O	O
average	NN	O	O
angle	NN	O	O
of	NN	O	O
about	NN	O	O
90	NN	O	O
degrees	NN	O	O
with	NN	O	O
helices	NN	O	B-protein
I	NN	O	I-protein
and	NN	O	I-protein
II	NN	O	I-protein
,	NN	O	O
and	NN	O	O
constitutes	NN	O	O
the	NN	O	O
other	NN	O	O
arm	NN	O	O
of	NN	O	O
the	NN	O	O
molecule	NN	O	O
.	NN	O	O

As	NN	O	O
in	NN	O	O
HMG1B	NN	O	B-protein
,	NN	O	O
the	NN	O	O
overall	NN	O	O
structure	NN	O	O
of	NN	O	O
the	NN	O	O
Sox-4	NN	O	B-protein
HMG	NN	O	B-protein
box	NN	O	I-protein
is	NN	O	O
L-shaped	NN	O	O
and	NN	O	O
is	NN	O	O
maintained	NN	O	O
by	NN	O	O
a	NN	O	O
cluster	NN	O	O
of	NN	O	O
conserved	NN	O	O
,	NN	O	O
mainly	NN	O	O
aromatic	NN	O	O
residues	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nuclear	NN	O	B-protein
factor-IL6	NN	O	I-protein
activates	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
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
I	NN	O	I-DNA
(	NN	O	O
PRE-I	NN	O	B-DNA
)	NN	O	O
is	NN	O	O
a	NN	O	O
strong	NN	O	O
enhancer	NN	O	B-DNA
element	NN	O	I-DNA
essential	NN	O	O
for	NN	O	O
expression	NN	O	O
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

To	NN	O	O
identify	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
PRE-I	NN	O	B-DNA
,	NN	O	O
we	NN	O	O
screened	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
expression	NN	O	I-DNA
library	NN	O	I-DNA
from	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
isolated	NN	O	O
a	NN	O	O
cDNA	NN	O	B-DNA
encoding	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	B-protein
NF	NN	O	I-protein
)	NN	O	I-protein
-IL6	NN	O	I-protein
(	NN	O	O
also	NN	O	O
known	NN	O	O
as	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
)	NN	O	O
.	NN	O	O

NF-IL6	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
found	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
in	NN	O	O
the	NN	O	O
mouse	NN	O	B-cell_line
Th2	NN	O	I-cell_line
clone	NN	O	I-cell_line
D10	NN	O	B-cell_line
,	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
Th1	NN	O	B-cell_line
clone	NN	O	I-cell_line
29	NN	O	I-cell_line
.	NN	O	O

rNF-IL6	NN	O	B-protein
expressed	NN	O	O
in	NN	O	O
bacteria	NN	O	O
was	NN	O	O
shown	NN	O	O
to	NN	O	O
specifically	NN	O	O
bind	NN	O	O
to	NN	O	O
PRE-I	NN	O	B-DNA
.	NN	O	O

PRE-I	NN	O	B-DNA
forms	NN	O	O
multiple	NN	O	B-protein
DNA-protein	NN	O	I-protein
complexes	NN	O	I-protein
with	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Some	NN	O	O
of	NN	O	O
these	NN	O	O
complexes	NN	O	O
were	NN	O	O
demonstrated	NN	O	O
to	NN	O	O
contain	NN	O	O
NF-IL6	NN	O	B-protein
by	NN	O	O
using	NN	O	O
anti-C/EBP	NN	O	B-protein
beta	NN	O	I-protein
Abs	NN	O	I-protein
.	NN	O	O

Overexpression	NN	O	O
of	NN	O	O
NF-IL6	NN	O	B-protein
enhanced	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
chloramphenicol	NN	O	B-DNA
acetyl	NN	O	I-DNA
transferase	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
PRE-I-thymidine	NN	O	B-protein
kinase	NN	O	I-protein
or	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
IL-4	NN	O	I-DNA
promoter	NN	O	I-DNA
more	NN	O	O
than	NN	O	O
10-fold	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Promoter	NN	O	O
deletion	NN	O	O
studies	NN	O	O
revealed	NN	O	O
two	NN	O	O
additional	NN	O	O
NF-IL6	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
located	NN	O	O
at	NN	O	O
positions	NN	O	O
-44	NN	O	O
to	NN	O	O
-36	NN	O	O
(	NN	O	O
C/EBP	NN	O	B-protein
proximal	NN	O	I-protein
)	NN	O	O
and	NN	O	O
-87	NN	O	O
to	NN	O	O
-79	NN	O	O
(	NN	O	O
C/EBP	NN	O	B-protein
medial	NN	O	I-protein
)	NN	O	O
,	NN	O	O
respectively	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
NF-IL6	NN	O	B-protein
is	NN	O	O
involved	NN	O	O
in	NN	O	O
transcriptional	NN	O	O
activation	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
in	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
an	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha-associated	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
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
and	NN	O	O
determination	NN	O	O
of	NN	O	O
its	NN	O	O
phosphorylation	NN	O	B-protein
sites	NN	O	I-protein
on	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	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
stored	NN	O	O
in	NN	O	O
the	NN	O	O
cytoplasm	NN	O	O
as	NN	O	O
an	NN	O	O
inactive	NN	O	O
form	NN	O	O
through	NN	O	O
interaction	NN	O	O
with	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
cells	NN	O	O
leads	NN	O	O
to	NN	O	O
a	NN	O	O
rapid	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
which	NN	O	O
is	NN	O	O
presumed	NN	O	O
to	NN	O	O
be	NN	O	O
important	NN	O	O
for	NN	O	O
the	NN	O	O
subsequent	NN	O	O
degradation	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
recently	NN	O	O
reported	NN	O	O
the	NN	O	O
establishment	NN	O	O
of	NN	O	O
a	NN	O	O
lipopolysaccharide	NN	O	O
(	NN	O	O
LPS	NN	O	O
)	NN	O	O
-dependent	NN	O	O
cell-free	NN	O	O
activation	NN	O	O
system	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
association	NN	O	O
with	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
.	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
identified	NN	O	O
a	NN	O	O
kinase	NN	O	B-protein
in	NN	O	O
cell	NN	O	O
extracts	NN	O	O
from	NN	O	O
the	NN	O	O
LPS-stimulated	NN	O	B-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
,	NN	O	O
THP-1	NN	O	B-cell_line
,	NN	O	O
that	NN	O	O
specifically	NN	O	O
binds	NN	O	O
and	NN	O	O
phosphorylates	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

LPS	NN	O	O
stimulation	NN	O	O
transiently	NN	O	O
enhanced	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
-bound	NN	O	B-protein
kinase	NN	O	I-protein
activity	NN	O	O
in	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Mutational	NN	O	O
analyses	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
and	NN	O	O
competition	NN	O	O
experiments	NN	O	O
with	NN	O	O
the	NN	O	O
synthetic	NN	O	O
peptides	NN	O	O
identified	NN	O	O
major	NN	O	O
phosphorylation	NN	O	B-protein
sites	NN	O	I-protein
by	NN	O	O
the	NN	O	O
bound	NN	O	O
kinase	NN	O	B-protein
as	NN	O	O
Ser	NN	O	O
and	NN	O	O
Thr	NN	O	O
residues	NN	O	O
in	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
acidic	NN	O	I-protein
domain	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
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
peptide	NN	O	O
,	NN	O	O
corresponding	NN	O	O
to	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-protein
acidic	NN	O	I-protein
domain	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
,	NN	O	O
blocked	NN	O	O
the	NN	O	O
LPS-induced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
inducible	NN	O	O
phosphorylation	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
a	NN	O	O
cell-free	NN	O	O
system	NN	O	O
using	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggested	NN	O	O
that	NN	O	O
the	NN	O	O
bound	NN	O	B-protein
kinase	NN	O	I-protein
is	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
LPS	NN	O	O
by	NN	O	O
inducing	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
C-terminal	NN	O	B-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
and	NN	O	O
subsequent	NN	O	O
dissociation	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
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	I-protein
complex	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Bik	NN	O	B-protein
,	NN	O	O
a	NN	O	O
novel	NN	O	O
death-inducing	NN	O	B-protein
protein	NN	O	I-protein
shares	NN	O	O
a	NN	O	O
distinct	NN	O	O
sequence	NN	O	O
motif	NN	O	O
with	NN	O	O
Bcl-2	NN	O	B-protein
family	NN	O	I-protein
proteins	NN	O	I-protein
and	NN	O	O
interacts	NN	O	O
with	NN	O	O
viral	NN	O	O
and	NN	O	O
cellular	NN	O	B-protein
survival-promoting	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

The	NN	O	O
survival-promoting	NN	O	O
activity	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
appears	NN	O	O
to	NN	O	O
be	NN	O	O
modulated	NN	O	O
by	NN	O	O
interactions	NN	O	O
between	NN	O	O
various	NN	O	O
cellular	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
identified	NN	O	O
a	NN	O	O
novel	NN	O	O
cellular	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
Bik	NN	O	B-protein
,	NN	O	O
that	NN	O	O
interacts	NN	O	O
with	NN	O	O
the	NN	O	O
cellular	NN	O	B-protein
survival-promoting	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
Bcl-2	NN	O	B-protein
and	NN	O	O
Bcl-xL	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
the	NN	O	O
viral	NN	O	B-protein
survival-promoting	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
Epstein	NN	O	B-protein
Barr	NN	O	I-protein
virus-BHRF1	NN	O	I-protein
and	NN	O	O
adenovirus	NN	O	B-protein
E1B-19	NN	O	I-protein
kDa	NN	O	I-protein
.	NN	O	O

In	NN	O	O
transient	NN	O	O
transfection	NN	O	O
assays	NN	O	O
,	NN	O	O
Bik	NN	O	B-protein
promotes	NN	O	O
cell	NN	O	O
death	NN	O	O
in	NN	O	O
a	NN	O	O
manner	NN	O	O
similar	NN	O	O
to	NN	O	O
the	NN	O	O
death-promoting	NN	O	B-protein
members	NN	O	I-protein
of	NN	O	O
the	NN	O	O
Bcl-2	NN	O	B-protein
family	NN	O	I-protein
,	NN	O	O
Bax	NN	O	B-protein
and	NN	O	O
Bak	NN	O	B-protein
.	NN	O	O

This	NN	O	O
death-promoting	NN	O	O
activity	NN	O	O
of	NN	O	O
Bik	NN	O	B-protein
can	NN	O	O
be	NN	O	O
suppressed	NN	O	O
by	NN	O	O
coexpression	NN	O	O
of	NN	O	O
Bcl-2	NN	O	B-protein
,	NN	O	O
Bcl-XL	NN	O	B-protein
,	NN	O	O
EBV-BHRF1	NN	O	B-protein
and	NN	O	O
E1B-19	NN	O	B-protein
kDa	NN	O	I-protein
proteins	NN	O	O
suggesting	NN	O	O
that	NN	O	O
Bik	NN	O	B-protein
may	NN	O	O
be	NN	O	O
a	NN	O	O
common	NN	O	O
target	NN	O	O
for	NN	O	O
both	NN	O	O
cellular	NN	O	B-protein
and	NN	O	I-protein
viral	NN	O	I-protein
anti-apoptotic	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

While	NN	O	O
Bik	NN	O	B-protein
does	NN	O	O
not	NN	O	O
show	NN	O	O
overt	NN	O	O
homology	NN	O	O
to	NN	O	O
the	NN	O	O
BH1	NN	O	B-protein
and	NN	O	O
BH2	NN	O	B-protein
conserved	NN	O	O
domains	NN	O	O
characteristic	NN	O	O
of	NN	O	O
the	NN	O	O
Bcl-2	NN	O	B-protein
family	NN	O	I-protein
,	NN	O	O
it	NN	O	O
does	NN	O	O
share	NN	O	O
a	NN	O	O
9	NN	O	O
amino	NN	O	B-protein
acid	NN	O	I-protein
domain	NN	O	I-protein
(	NN	O	O
BH3	NN	O	B-protein
)	NN	O	O
with	NN	O	O
Bax	NN	O	B-protein
and	NN	O	O
Bak	NN	O	B-protein
which	NN	O	O
may	NN	O	O
be	NN	O	O
a	NN	O	O
critical	NN	O	O
determinant	NN	O	O
for	NN	O	O
the	NN	O	O
death-promoting	NN	O	O
activity	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
human	NN	O	B-DNA
TCF-1	NN	O	I-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
DNA-binding	NN	O	I-protein
protein	NN	O	I-protein
uniquely	NN	O	O
expressed	NN	O	O
in	NN	O	O
normal	NN	O	B-cell_line
and	NN	O	I-cell_line
neoplastic	NN	O	I-cell_line
T-lineage	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
TCF-1	NN	O	B-DNA
gene	NN	O	I-DNA
encodes	NN	O	O
a	NN	O	O
putative	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
with	NN	O	O
affinity	NN	O	O
for	NN	O	O
a	NN	O	O
sequence	NN	O	O
motif	NN	O	O
occurring	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
T-cell	NN	O	B-DNA
enhancers	NN	O	I-DNA
.	NN	O	O

TCF-1	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
originally	NN	O	O
found	NN	O	O
to	NN	O	O
be	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
T	NN	O	O
cell-specific	NN	O	O
fashion	NN	O	O
within	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
and	NN	O	I-cell_line
mouse	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
expression	NN	O	O
reportedly	NN	O	O
occurs	NN	O	O
in	NN	O	O
multiple	NN	O	O
nonlymphoid	NN	O	O
tissues	NN	O	O
during	NN	O	O
murine	NN	O	O
embryogenesis	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
now	NN	O	O
raised	NN	O	O
a	NN	O	O
monoclonal	NN	O	B-protein
antibody	NN	O	I-protein
to	NN	O	O
document	NN	O	O
expression	NN	O	O
and	NN	O	O
biochemistry	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-protein
TCF-1	NN	O	I-protein
protein	NN	O	I-protein
.	NN	O	O

As	NN	O	O
expected	NN	O	O
,	NN	O	O
the	NN	O	O
TCF-1	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
detectable	NN	O	O
only	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
T	NN	O	I-cell_line
lineage	NN	O	I-cell_line
.	NN	O	O

Its	NN	O	O
expression	NN	O	O
was	NN	O	O
always	NN	O	O
restricted	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Immunohistochemistry	NN	O	O
on	NN	O	O
a	NN	O	O
panel	NN	O	O
of	NN	O	O
human	NN	O	O
tissues	NN	O	O
revealed	NN	O	O
that	NN	O	O
the	NN	O	O
TCF-1	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
found	NN	O	O
exclusively	NN	O	O
in	NN	O	O
thymocytes	NN	O	O
and	NN	O	O
in	NN	O	O
CD3+	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
in	NN	O	O
peripheral	NN	O	O
lymphoid	NN	O	O
tissues	NN	O	O
.	NN	O	O

Western	NN	O	O
blotting	NN	O	O
yielded	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
bands	NN	O	O
ranging	NN	O	O
from	NN	O	O
25	NN	O	O
kD	NN	O	O
to	NN	O	O
55	NN	O	O
kD	NN	O	O
,	NN	O	O
resulting	NN	O	O
from	NN	O	O
extensive	NN	O	O
alternative	NN	O	O
splicing	NN	O	O
.	NN	O	O

The	NN	O	O
TCF-1	NN	O	B-protein
protein	NN	O	O
was	NN	O	O
detectable	NN	O	O
in	NN	O	O
all	NN	O	O
samples	NN	O	O
of	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
22	NN	O	O
T-cell	NN	O	B-cell_type
malignancies	NN	O	I-cell_type
of	NN	O	O
various	NN	O	O
stages	NN	O	O
of	NN	O	O
maturation	NN	O	O
,	NN	O	O
but	NN	O	O
was	NN	O	O
absent	NN	O	O
from	NN	O	O
a	NN	O	O
large	NN	O	O
number	NN	O	O
of	NN	O	O
other	NN	O	O
hematologic	NN	O	B-cell_type
neoplasms	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
observations	NN	O	O
imply	NN	O	O
a	NN	O	O
T	NN	O	O
cell-specific	NN	O	O
function	NN	O	O
for	NN	O	O
TCF-1	NN	O	B-protein
,	NN	O	O
a	NN	O	O
notion	NN	O	O
corroborated	NN	O	O
by	NN	O	O
recent	NN	O	O
observations	NN	O	O
on	NN	O	O
Tcf-1	NN	O	O
knock-out	NN	O	O
mice	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
these	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
nuclear	NN	O	O
TCF-1	NN	O	B-protein
expression	NN	O	O
can	NN	O	O
serve	NN	O	O
as	NN	O	O
a	NN	O	O
pan-T-lineage	NN	O	O
marker	NN	O	O
in	NN	O	O
the	NN	O	O
diagnosis	NN	O	O
of	NN	O	O
lymphoid	NN	O	O
malignancies	NN	O	O
.	NN	O	O

-DOCSTART-	O

Cross-linking	NN	O	O
of	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
receptors	NN	O	I-protein
activates	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
transcription	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
.	NN	O	O

Elevation	NN	O	O
of	NN	O	O
the	NN	O	O
levels	NN	O	O
of	NN	O	O
circulating	NN	O	B-protein
immune	NN	O	I-protein
complexes	NN	O	I-protein
frequently	NN	O	O
accompanies	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
and	NN	O	O
is	NN	O	O
a	NN	O	O
prognostic	NN	O	O
indicator	NN	O	O
of	NN	O	O
clinical	NN	O	O
progression	NN	O	O
from	NN	O	O
asymptomatic	NN	O	O
infection	NN	O	O
to	NN	O	O
AIDS	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
cross-linking	NN	O	O
of	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RI	NN	O	I-protein
or	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
RII	NN	O	I-protein
by	NN	O	O
adherent	NN	O	B-protein
human	NN	O	I-protein
IgG	NN	O	I-protein
or	NN	O	O
by	NN	O	O
specific	NN	O	O
anti-Fc	NN	O	B-protein
gamma	NN	O	I-protein
R	NN	O	I-protein
mAb	NN	O	I-protein
activates	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
expression	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
BF24	NN	O	B-cell_line
and	NN	O	O
increased	NN	O	O
HIV	NN	O	B-RNA
RNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
from	NN	O	O
HIV	NN	O	O
infected	NN	O	O
patients	NN	O	O
as	NN	O	O
assayed	NN	O	O
by	NN	O	O
reverse	NN	O	O
transcription-PCR	NN	O	O
.	NN	O	O

In	NN	O	O
THP-1	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
R	NN	O	I-protein
cross-linking	NN	O	O
induced	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
known	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
regulatory	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
of	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
to	NN	O	O
activate	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
.	NN	O	O
Anti-TNF-alpha	NN	O	B-protein
antibody	NN	O	I-protein
but	NN	O	O
not	NN	O	O
anti-IL-1	NN	O	B-protein
beta	NN	O	I-protein
antibody	NN	O	I-protein
strongly	NN	O	O
inhibited	NN	O	O
both	NN	O	O
the	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
and	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
by	NN	O	O
Fc	NN	O	B-protein
gamma	NN	O	I-protein
R	NN	O	I-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
Fc	NN	O	B-protein
gamma	NN	O	I-protein
R	NN	O	I-protein
can	NN	O	O
mediate	NN	O	O
a	NN	O	O
TNF-alpha	NN	O	B-protein
-dependent	NN	O	O
induction	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
immune	NN	O	B-protein
complexes	NN	O	I-protein
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
the	NN	O	O
pathophysiology	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
infection	NN	O	O
by	NN	O	O
augmenting	NN	O	O
viral	NN	O	O
replication	NN	O	O
in	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

-DOCSTART-	O

Signalling	NN	O	O
via	NN	O	O
CD28	NN	O	B-protein
of	NN	O	O
human	NN	O	B-cell_line
naive	NN	O	I-cell_line
neonatal	NN	O	I-cell_line
T	NN	O	I-cell_line
lymphocytes	NN	O	I-cell_line
.	NN	O	O

Accessory	NN	O	O
molecules	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
development	NN	O	O
of	NN	O	O
the	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
to	NN	O	O
antigenic	NN	O	O
challenge	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
CD28	NN	O	B-protein
in	NN	O	O
modulating	NN	O	O
the	NN	O	O
'naive	NN	O	O
'	NN	O	O
neonatal	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
to	NN	O	O
anti-CD2	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
.	NN	O	O

To	NN	O	O
compare	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
CD28	NN	O	B-protein
,	NN	O	O
neonatal	NN	O	B-cell_type
and	NN	O	O
adult	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
a	NN	O	O
pair	NN	O	O
of	NN	O	O
mitogenic	NN	O	B-protein
anti-CD2	NN	O	I-protein
antibodies	NN	O	I-protein
in	NN	O	O
the	NN	O	O
presence	NN	O	O
or	NN	O	O
absence	NN	O	O
of	NN	O	O
anti-CD28	NN	O	B-protein
MoAb	NN	O	I-protein
.	NN	O	O

With	NN	O	O
anti-CD2	NN	O	B-protein
alone	NN	O	O
,	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
proliferated	NN	O	O
slightly	NN	O	O
but	NN	O	O
produced	NN	O	O
no	NN	O	O
detectable	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
whereas	NN	O	O
adult	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
proliferated	NN	O	O
vigorously	NN	O	O
,	NN	O	O
with	NN	O	O
significant	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Costimulation	NN	O	O
with	NN	O	O
anti-CD28	NN	O	B-protein
MoAb	NN	O	I-protein
greatly	NN	O	O
enhanced	NN	O	O
the	NN	O	O
proliferative	NN	O	O
response	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
levels	NN	O	O
equivalent	NN	O	O
to	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
whereas	NN	O	O
adult	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
showed	NN	O	O
only	NN	O	O
slight	NN	O	O
increases	NN	O	O
.	NN	O	O

Although	NN	O	O
IL-2	NN	O	B-protein
secretion	NN	O	O
was	NN	O	O
increased	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
anti-CD28	NN	O	B-protein
MoAb	NN	O	I-protein
,	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
cell	NN	O	I-cell_type
IL-2	NN	O	B-protein
production	NN	O	O
remained	NN	O	O
lower	NN	O	O
than	NN	O	O
in	NN	O	O
adults	NN	O	O
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
enhancement	NN	O	O
of	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
in	NN	O	O
neonates	NN	O	O
was	NN	O	O
similar	NN	O	O
to	NN	O	O
adult	NN	O	O
levels	NN	O	O
.	NN	O	O

Anti-	NN	O	O
CD28	NN	O	B-protein
MoAb	NN	O	I-protein
costimulation	NN	O	O
increased	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
levels	NN	O	O
in	NN	O	O
neonates	NN	O	O
,	NN	O	O
albeit	NN	O	O
to	NN	O	O
levels	NN	O	O
lower	NN	O	O
than	NN	O	O
that	NN	O	O
of	NN	O	O
adults	NN	O	O
.	NN	O	O

The	NN	O	O
cellular	NN	O	O
mechanism	NN	O	O
governing	NN	O	O
the	NN	O	O
diminished	NN	O	O
proliferative	NN	O	O
response	NN	O	O
of	NN	O	O
neonatal	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
to	NN	O	O
anti-CD2	NN	O	B-protein
may	NN	O	O
therefore	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
decreased	NN	O	O
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
induction	NN	O	O
,	NN	O	O
reduced	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
deficient	NN	O	O
IL-2	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Although	NN	O	O
anti-CD28	NN	O	B-protein
MoAb	NN	O	I-protein
costimulation	NN	O	O
enhances	NN	O	O
all	NN	O	O
of	NN	O	O
the	NN	O	O
above	NN	O	O
signals	NN	O	O
,	NN	O	O
NF	NN	O	O
kappa	NN	O	O
B	NN	O	O
and	NN	O	O
IL-2	NN	O	O
levels	NN	O	O
remain	NN	O	O
lower	NN	O	O
than	NN	O	O
in	NN	O	O
adults	NN	O	O
,	NN	O	O
suggesting	NN	O	O
the	NN	O	O
need	NN	O	O
for	NN	O	O
further	NN	O	O
activation	NN	O	O
requirements	NN	O	O
in	NN	O	O
the	NN	O	O
neonate	NN	O	O
.	NN	O	O

-DOCSTART-	O

TCL1	NN	O	O
oncogene	NN	O	O
activation	NN	O	O
in	NN	O	O
preleukemic	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
case	NN	O	O
of	NN	O	O
ataxia-telangiectasia	NN	O	O
.	NN	O	O

The	NN	O	O
TCL1	NN	O	B-DNA
oncogene	NN	O	I-DNA
on	NN	O	O
human	NN	O	B-DNA
chromosome	NN	O	I-DNA
14q32.1	NN	O	B-DNA
is	NN	O	O
involved	NN	O	O
in	NN	O	O
chromosome	NN	O	O
translocations	NN	O	O
[	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
14	NN	O	I-DNA
;	NN	O	I-DNA
14	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q11	NN	O	I-DNA
;	NN	O	I-DNA
q32.1	NN	O	I-DNA
)	NN	O	I-DNA
and	NN	O	O
t	NN	O	B-DNA
(	NN	O	I-DNA
7	NN	O	I-DNA
;	NN	O	I-DNA
14	NN	O	I-DNA
)	NN	O	I-DNA
(	NN	O	I-DNA
q35	NN	O	I-DNA
;	NN	O	I-DNA
q32.1	NN	O	I-DNA
)	NN	O	I-DNA
]	NN	O	O
and	NN	O	O
inversions	NN	O	O
[	NN	O	O
inv14	NN	O	B-DNA
(	NN	O	I-DNA
q11	NN	O	I-DNA
;	NN	O	I-DNA
q32.1	NN	O	I-DNA
)	NN	O	I-DNA
]	NN	O	O
with	NN	O	O
TCR	NN	O	B-DNA
alpha/beta	NN	O	I-DNA
loci	NN	O	I-DNA
in	NN	O	O
T-cell	NN	O	O
leukemias	NN	O	O
,	NN	O	O
such	NN	O	O
as	NN	O	O
T-prolymphocytic	NN	O	O
(	NN	O	O
T-PLL	NN	O	O
)	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
also	NN	O	O
involved	NN	O	O
in	NN	O	O
T-	NN	O	O
acute	NN	O	O
and-	NN	O	O
chronic	NN	O	O
leukemias	NN	O	O
arising	NN	O	O
in	NN	O	O
cases	NN	O	O
of	NN	O	O
ataxia-telangiectasia	NN	O	O
(	NN	O	O
AT	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
immunodeficiency	NN	O	O
syndrome	NN	O	O
.	NN	O	O

Similar	NN	O	O
chromosomal	NN	O	O
rearrangements	NN	O	O
occur	NN	O	O
also	NN	O	O
in	NN	O	O
the	NN	O	O
clonally	NN	O	O
expanded	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
AT	NN	O	O
patients	NN	O	O
before	NN	O	O
the	NN	O	O
appearance	NN	O	O
of	NN	O	O
the	NN	O	O
overt	NN	O	O
leukemia	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
TCL1	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	B-protein
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
from	NN	O	O
four	NN	O	O
AT	NN	O	O
cases	NN	O	O
and	NN	O	O
from	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
the	NN	O	O
TCL1	NN	O	B-DNA
gene	NN	O	I-DNA
was	NN	O	O
overexpressed	NN	O	O
in	NN	O	O
the	NN	O	O
PBLs	NN	O	B-cell_type
of	NN	O	O
an	NN	O	O
AT	NN	O	O
patient	NN	O	O
with	NN	O	O
a	NN	O	O
large	NN	O	B-cell_line
clonal	NN	O	I-cell_line
T-cell	NN	O	I-cell_line
population	NN	O	I-cell_line
exhibiting	NN	O	O
the	NN	O	O
t	NN	O	O
(	NN	O	O
14	NN	O	O
;	NN	O	O
14	NN	O	O
)	NN	O	O
translocation	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
the	NN	O	O
lymphocytes	NN	O	B-cell_type
of	NN	O	O
the	NN	O	O
other	NN	O	O
cases	NN	O	O
.	NN	O	O

Fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
of	NN	O	O
the	NN	O	O
TCL1	NN	O	B-DNA
genomic	NN	O	I-DNA
locus	NN	O	I-DNA
to	NN	O	O
lymphocyte	NN	O	O
metaphases	NN	O	O
from	NN	O	O
the	NN	O	O
AT	NN	O	O
patient	NN	O	O
with	NN	O	O
the	NN	O	O
T-cell	NN	O	O
clonal	NN	O	O
expansion	NN	O	O
showed	NN	O	O
that	NN	O	O
the	NN	O	O
breakpoint	NN	O	O
of	NN	O	O
the	NN	O	O
t	NN	O	O
(	NN	O	O
14	NN	O	O
;	NN	O	O
14	NN	O	O
)	NN	O	O
translocation	NN	O	O
lies	NN	O	O
within	NN	O	O
the	NN	O	O
TCL1	NN	O	B-DNA
locus	NN	O	I-DNA
and	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
an	NN	O	O
inverted	NN	O	O
duplication	NN	O	O
of	NN	O	O
the	NN	O	O
distal	NN	O	B-DNA
part	NN	O	I-DNA
of	NN	O	O
chromosome	NN	O	B-DNA
14	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
TCL1	NN	O	B-DNA
is	NN	O	O
activated	NN	O	O
in	NN	O	O
preleukemic	NN	O	B-cell_line
clonal	NN	O	I-cell_line
cells	NN	O	I-cell_line
as	NN	O	O
a	NN	O	O
consequence	NN	O	O
of	NN	O	O
chromosome	NN	O	O
translocation	NN	O	O
involving	NN	O	O
sequences	NN	O	O
from	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
locus	NN	O	I-DNA
at	NN	O	O
14q11	NN	O	B-DNA
.	NN	O	O

Deregulation	NN	O	O
of	NN	O	O
TCL1	NN	O	B-DNA
is	NN	O	O
the	NN	O	O
first	NN	O	O
event	NN	O	O
in	NN	O	O
the	NN	O	O
initiation	NN	O	O
of	NN	O	O
malignancy	NN	O	O
in	NN	O	O
these	NN	O	O
types	NN	O	O
of	NN	O	O
leukemias	NN	O	O
and	NN	O	O
represents	NN	O	O
a	NN	O	O
potential	NN	O	O
tool	NN	O	O
for	NN	O	O
clinical	NN	O	O
evaluation	NN	O	O
.	NN	O	O

-DOCSTART-	O

C/EBP	NN	O	B-protein
proteins	NN	O	I-protein
activate	NN	O	O
transcription	NN	O	O
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
1	NN	O	I-DNA
long	NN	O	I-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
in	NN	O	O
macrophages/	NN	O	O
monocytes	NN	O	B-cell_type
.	NN	O	O

Three	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
C/EBP	NN	O	B-protein
proteins	NN	O	I-protein
are	NN	O	O
found	NN	O	O
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
(	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
LTR	NN	O	B-DNA
)	NN	O	O
(	NN	O	O
V.M.	NN	O	O
Tesmer	NN	O	O
,	NN	O	O
A.Rajadhyaksha	NN	O	O
,	NN	O	O
J.Babin	NN	O	O
,	NN	O	O
and	NN	O	O
M.Bina	NN	O	O
,	NN	O	O
Proc.Natl.Acad.Sci.	NN	O	O
USA	NN	O	O
90	NN	O	O
:	NN	O	O
7298-7302	NN	O	O
,	NN	O	O
1993	NN	O	O
)	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
determined	NN	O	O
the	NN	O	O
functional	NN	O	O
role	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
C/EBP	NN	O	B-DNA
sites	NN	O	I-DNA
in	NN	O	O
regulating	NN	O	O
transcription	NN	O	O
from	NN	O	O
the	NN	O	O
HIV-	NN	O	B-DNA
1	NN	O	I-DNA
LTR	NN	O	I-DNA
in	NN	O	O
monocytes/macrophages	NN	O	B-cell_type
.	NN	O	O

Inhibition	NN	O	O
of	NN	O	O
endogenous	NN	O	B-protein
C/EBP	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
using	NN	O	O
either	NN	O	O
an	NN	O	O
excess	NN	O	O
of	NN	O	O
C/EBP	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
or	NN	O	O
a	NN	O	O
trans-	NN	O	B-DNA
dominant	NN	O	I-DNA
negative	NN	O	I-DNA
inhibitor	NN	O	I-DNA
,	NN	O	O
demonstrated	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
basal	NN	O	O
and	NN	O	O
activated	NN	O	O
levels	NN	O	O
of	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
transcription	NN	O	O
in	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

Northern	NN	O	O
(	NN	O	O
RNA	NN	O	O
)	NN	O	O
blots	NN	O	O
and	NN	O	O
binding	NN	O	O
assays	NN	O	O
showed	NN	O	O
that	NN	O	O
NF-IL6	NN	O	B-protein
is	NN	O	O
the	NN	O	O
only	NN	O	O
known	NN	O	O
C/EBP	NN	O	B-protein
family	NN	O	I-protein
member	NN	O	I-protein
which	NN	O	O
is	NN	O	O
increased	NN	O	O
when	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
activated	NN	O	O
.	NN	O	O

Mutational	NN	O	O
analyses	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
LTR	NN	O	I-DNA
showed	NN	O	O
that	NN	O	O
one	NN	O	O
C/EBP	NN	O	B-DNA
site	NN	O	I-DNA
is	NN	O	O
required	NN	O	O
for	NN	O	O
normal	NN	O	O
LTR	NN	O	B-DNA
transcription	NN	O	O
both	NN	O	O
before	NN	O	O
and	NN	O	O
after	NN	O	O
cellular	NN	O	O
activation	NN	O	O
and	NN	O	O
that	NN	O	O
the	NN	O	O
two	NN	O	O
3	NN	O	O
'	NN	O	O
C/EBP	NN	O	B-DNA
sites	NN	O	I-DNA
are	NN	O	O
functionally	NN	O	O
equivalent	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
transcription	NN	O	O
from	NN	O	O
crippled	NN	O	B-DNA
HIV-1	NN	O	I-DNA
LTRs	NN	O	I-DNA
lacking	NN	O	O
C/EBP	NN	O	B-DNA
sites	NN	O	I-DNA
can	NN	O	O
still	NN	O	O
be	NN	O	O
induced	NN	O	O
following	NN	O	O
activation	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Several	NN	O	O
models	NN	O	O
are	NN	O	O
suggested	NN	O	O
for	NN	O	O
how	NN	O	O
elevated	NN	O	O
NF-IL6	NN	O	B-protein
may	NN	O	O
participate	NN	O	O
in	NN	O	O
an	NN	O	O
autostimulatory	NN	O	O
loop	NN	O	O
involving	NN	O	O
HIV	NN	O	O
infection	NN	O	O
,	NN	O	O
macrophage	NN	O	O
activation	NN	O	O
,	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
HIV	NN	O	O
replication	NN	O	O
.	NN	O	O

-DOCSTART-	O

Human	NN	O	O
immunodeficiency	NN	O	O
virus	NN	O	O
type-2	NN	O	O
gene	NN	O	O
expression	NN	O	O
:	NN	O	O
two	NN	O	O
enhancers	NN	O	B-DNA
and	NN	O	O
their	NN	O	O
activation	NN	O	O
by	NN	O	O
T-cell	NN	O	B-cell_type
activators	NN	O	O
.	NN	O	O

The	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
may	NN	O	O
include	NN	O	O
a	NN	O	O
spectrum	NN	O	O
of	NN	O	O
retroviruses	NN	O	O
with	NN	O	O
varying	NN	O	O
potential	NN	O	O
to	NN	O	O
infect	NN	O	O
their	NN	O	O
host	NN	O	O
,	NN	O	O
undergo	NN	O	O
long	NN	O	O
periods	NN	O	O
of	NN	O	O
latent	NN	O	O
infection	NN	O	O
,	NN	O	O
and	NN	O	O
induce	NN	O	O
pathology	NN	O	O
.	NN	O	O

Since	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
viruses	NN	O	O
is	NN	O	O
in	NN	O	O
large	NN	O	O
part	NN	O	O
regulated	NN	O	O
by	NN	O	O
the	NN	O	O
sequence	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
their	NN	O	O
long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeats	NN	O	I-DNA
(	NN	O	O
LTRs	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
this	NN	O	O
study	NN	O	O
was	NN	O	O
directed	NN	O	O
to	NN	O	O
an	NN	O	O
analysis	NN	O	O
of	NN	O	O
the	NN	O	O
regulatory	NN	O	B-DNA
elements	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
HIV-2	NN	O	B-DNA
LTR	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
HIV-2	NN	O	B-DNA
LTR	NN	O	I-DNA
was	NN	O	O
found	NN	O	O
to	NN	O	O
contain	NN	O	O
two	NN	O	O
enhancers	NN	O	B-DNA
.	NN	O	O

One	NN	O	O
of	NN	O	O
these	NN	O	O
enhancers	NN	O	B-DNA
is	NN	O	O
,	NN	O	O
in	NN	O	O
part	NN	O	O
,	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
enhancer	NN	O	B-DNA
in	NN	O	O
HIV-1	NN	O	O
is	NN	O	O
the	NN	O	O
T-cell	NN	O	B-DNA
activation	NN	O	I-DNA
response	NN	O	I-DNA
element	NN	O	I-DNA
;	NN	O	O
in	NN	O	O
HIV-2	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
the	NN	O	O
second	NN	O	B-DNA
enhancer	NN	O	I-DNA
that	NN	O	O
is	NN	O	O
mainly	NN	O	O
responsible	NN	O	O
for	NN	O	O
activation	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
T-cell	NN	O	O
activators	NN	O	O
.	NN	O	O

The	NN	O	O
second	NN	O	O
enhancer	NN	O	B-DNA
interacts	NN	O	O
with	NN	O	O
two	NN	O	O
nuclear	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
85	NN	O	O
kD	NN	O	O
and	NN	O	O
27	NN	O	O
kD	NN	O	O
mobility	NN	O	O
)	NN	O	O
that	NN	O	O
appear	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
optimal	NN	O	O
enhancer	NN	O	O
function	NN	O	O
and	NN	O	O
activation	NN	O	O
.	NN	O	O

Observations	NN	O	O
such	NN	O	O
as	NN	O	O
these	NN	O	O
encourage	NN	O	O
the	NN	O	O
speculation	NN	O	O
that	NN	O	O
there	NN	O	O
may	NN	O	O
be	NN	O	O
subtle	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
and	NN	O	O
HIV-2	NN	O	O
expression	NN	O	O
that	NN	O	O
may	NN	O	O
be	NN	O	O
relevant	NN	O	O
to	NN	O	O
the	NN	O	O
possible	NN	O	O
longer	NN	O	O
latency	NN	O	O
and	NN	O	O
reduced	NN	O	O
pathogenicity	NN	O	O
of	NN	O	O
HIV-2	NN	O	O
.	NN	O	O

-DOCSTART-	O

NF-M	NN	O	B-protein
(	NN	O	O
chicken	NN	O	B-protein
C/EBP	NN	O	I-protein
beta	NN	O	I-protein
)	NN	O	O
induces	NN	O	O
eosinophilic	NN	O	O
differentiation	NN	O	O
and	NN	O	O
apoptosis	NN	O	O
in	NN	O	O
a	NN	O	O
hematopoietic	NN	O	B-cell_line
progenitor	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
.	NN	O	O

CAAT/enhancer	NN	O	B-protein
binding	NN	O	I-protein
proteins	NN	O	I-protein
(	NN	O	O
C/EBPs	NN	O	B-protein
)	NN	O	O
are	NN	O	O
transcriptional	NN	O	O
activators	NN	O	O
implicated	NN	O	O
in	NN	O	O
the	NN	O	O
differentiation	NN	O	O
processes	NN	O	O
of	NN	O	O
various	NN	O	O
cell	NN	O	O
lineages	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
earlier	NN	O	O
that	NN	O	O
NF-M	NN	O	B-protein
,	NN	O	O
the	NN	O	O
chicken	NN	O	B-protein
homolog	NN	O	I-protein
of	NN	O	O
C/EBP	NN	O	B-protein
beta	NN	O	I-protein
,	NN	O	O
is	NN	O	O
specifically	NN	O	O
expressed	NN	O	O
in	NN	O	O
myelomonocytic	NN	O	B-cell_type
and	NN	O	I-cell_type
eosinophilic	NN	O	I-cell_type
cells	NN	O	I-cell_type
of	NN	O	O
the	NN	O	O
hematopoietic	NN	O	O
system	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
NF-M	NN	O	B-protein
in	NN	O	O
hematopoietic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lineage	NN	O	I-cell_line
commitment	NN	O	O
,	NN	O	O
we	NN	O	O
constructed	NN	O	O
a	NN	O	O
conditional	NN	O	O
form	NN	O	O
of	NN	O	O
the	NN	O	O
protein	NN	O	O
by	NN	O	O
fusing	NN	O	O
it	NN	O	O
to	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
human	NN	O	B-protein
estrogen	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

This	NN	O	O
construct	NN	O	O
was	NN	O	O
stably	NN	O	O
expressed	NN	O	O
in	NN	O	O
a	NN	O	O
multipotent	NN	O	B-cell_line
progenitor	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
transformed	NN	O	O
by	NN	O	O
the	NN	O	O
Myb-Ets	NN	O	B-protein
oncoprotein	NN	O	I-protein
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	NN	O	O
that	NN	O	O
both	NN	O	O
NF-M-dependent	NN	O	B-DNA
promoter	NN	O	I-DNA
constructs	NN	O	I-DNA
and	NN	O	O
resident	NN	O	O
genes	NN	O	O
could	NN	O	O
be	NN	O	O
activated	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
beta-estradiol	NN	O	O
to	NN	O	O
the	NN	O	O
NF-M-estrogen	NN	O	B-cell_line
receptor	NN	O	I-cell_line
expressing	NN	O	I-cell_line
progenitors	NN	O	I-cell_line
.	NN	O	O

At	NN	O	O
the	NN	O	O
same	NN	O	O
time	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
a	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
progenitor-specific	NN	O	B-protein
surface	NN	O	I-protein
markers	NN	O	I-protein
and	NN	O	O
the	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
differentiation	NN	O	B-protein
markers	NN	O	I-protein
restricted	NN	O	O
to	NN	O	O
the	NN	O	O
eosinophil	NN	O	B-cell_type
and	NN	O	O
myeloid	NN	O	B-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
differentiation	NN	O	O
,	NN	O	O
cell	NN	O	O
death	NN	O	O
was	NN	O	O
induced	NN	O	O
with	NN	O	O
typical	NN	O	O
apoptotic	NN	O	O
features	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
suggest	NN	O	O
that	NN	O	O
NF-M	NN	O	B-protein
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
commitment	NN	O	O
along	NN	O	O
the	NN	O	O
eosinophil	NN	O	B-cell_type
lineage	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
apoptosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Prolactin	NN	O	B-protein
and	NN	O	O
interleukin-2	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
signal	NN	O	O
through	NN	O	O
a	NN	O	O
MGF-STAT5-like	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
.	NN	O	O

The	NN	O	O
cell	NN	O	O
surface	NN	O	O
receptors	NN	O	O
for	NN	O	O
PRL	NN	O	B-protein
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
structurally	NN	O	O
distinct	NN	O	O
,	NN	O	O
but	NN	O	O
share	NN	O	O
regulatory	NN	O	O
tasks	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

They	NN	O	O
can	NN	O	O
stimulate	NN	O	O
proliferation	NN	O	O
and	NN	O	O
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
over-lapping	NN	O	O
sets	NN	O	O
of	NN	O	O
genes	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

PRL	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
receptor	NN	O	O
activation	NN	O	O
are	NN	O	O
both	NN	O	O
linked	NN	O	O
to	NN	O	O
the	NN	O	O
Jak/Stat	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
)	NN	O	O
pathway	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
PRL	NN	O	B-protein
and	NN	O	O
IL-2	NN	O	B-protein
to	NN	O	O
activate	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
in	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

The	NN	O	O
DNA	NN	O	O
binding	NN	O	O
specificities	NN	O	O
,	NN	O	O
the	NN	O	O
reactivities	NN	O	O
toward	NN	O	O
Stat-specific	NN	O	O
antisera	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
mol	NN	O	O
wt	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
-and	NN	O	O
PRL-induced	NN	O	B-protein
DNA-binding	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
Nb2	NN	O	B-cell_line
and	NN	O	I-cell_line
C196	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
were	NN	O	O
investigated	NN	O	O
.	NN	O	O

A	NN	O	O
comparison	NN	O	O
with	NN	O	O
the	NN	O	O
Stat	NN	O	B-protein
proteins	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
interferon-gamma	NN	O	B-protein
,	NN	O	O
PRL	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL-6	NN	O	B-protein
in	NN	O	O
T47D	NN	O	B-cell_line
mammary	NN	O	I-cell_line
tumor	NN	O	I-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
made	NN	O	O
.	NN	O	O

We	NN	O	O
found	NN	O	O
that	NN	O	O
these	NN	O	O
parameters	NN	O	O
were	NN	O	O
indistinguishable	NN	O	O
for	NN	O	O
one	NN	O	O
of	NN	O	O
the	NN	O	O
PRL-	NN	O	B-protein
and	NN	O	I-protein
IL-2-induced	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

A	NN	O	O
transcription	NN	O	O
factor	NN	O	O
closely	NN	O	O
related	NN	O	O
to	NN	O	O
mammary	NN	O	B-protein
gland	NN	O	I-protein
factor-Stat5	NN	O	I-protein
is	NN	O	O
rapidly	NN	O	O
activated	NN	O	O
upon	NN	O	O
interaction	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
and	NN	O	O
PRL	NN	O	B-protein
with	NN	O	O
their	NN	O	O
respective	NN	O	O
receptors	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
a	NN	O	O
second	NN	O	O
protein	NN	O	O
related	NN	O	O
to	NN	O	O
Stat1	NN	O	B-protein
was	NN	O	O
also	NN	O	O
observed	NN	O	O
.	NN	O	O

Our	NN	O	O
results	NN	O	O
emphasize	NN	O	O
the	NN	O	O
role	NN	O	O
of	NN	O	O
PRL	NN	O	B-protein
as	NN	O	O
a	NN	O	O
regulator	NN	O	O
of	NN	O	O
the	NN	O	O
immune	NN	O	O
response	NN	O	O
and	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
Stat	NN	O	B-protein
factors	NN	O	I-protein
mammary	NN	O	B-protein
gland	NN	O	I-protein
factor-Stat5	NN	O	I-protein
and	NN	O	O
Stat1	NN	O	B-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
gene	NN	O	O
expression	NN	O	O
during	NN	O	O
T	NN	O	O
cell	NN	O	O
development	NN	O	O
.	NN	O	O

-DOCSTART-	O

ETS1	NN	O	B-protein
transactivates	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
GM-CSF	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
stimulated	NN	O	O
with	NN	O	O
PMA	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
helper	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-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
whose	NN	O	O
increased	NN	O	O
expression	NN	O	O
results	NN	O	O
partly	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
NF	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
,	NN	O	O
AP-1	NN	O	B-protein
and	NN	O	O
ETS-like	NN	O	B-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
which	NN	O	O
are	NN	O	O
important	NN	O	O
for	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
following	NN	O	O
PMA	NN	O	O
and	NN	O	O
ionomycin	NN	O	O
stimulation	NN	O	O
.	NN	O	O

A	NN	O	O
number	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
transcription	NN	O	B-protein
factors	NN	O	I-protein
are	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
including	NN	O	O
ETS1	NN	O	B-protein
and	NN	O	O
ELF1	NN	O	B-protein
.	NN	O	O

Here	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
these	NN	O	O
factors	NN	O	O
to	NN	O	O
interact	NN	O	O
with	NN	O	O
a	NN	O	O
site	NN	O	O
(	NN	O	O
GM5	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
CLE0	NN	O	B-DNA
element	NN	O	I-DNA
,	NN	O	O
-47	NN	O	B-DNA
to	NN	O	I-DNA
-40	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
transcription	NN	O	I-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Exogenous	NN	O	O
ETS1	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
ELF1	NN	O	B-protein
,	NN	O	O
can	NN	O	O
transactivate	NN	O	O
GM-CSF	NN	O	B-protein
,	NN	O	O
through	NN	O	O
the	NN	O	O
GM5	NN	O	B-DNA
site	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
PMA/ionomycin	NN	O	O
dependent	NN	O	O
manner	NN	O	O
.	NN	O	O

Other	NN	O	O
unidentified	NN	O	O
ETS-like	NN	O	B-protein
factors	NN	O	I-protein
present	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
are	NN	O	O
also	NN	O	O
capable	NN	O	O
of	NN	O	O
binding	NN	O	O
GM5	NN	O	B-DNA
.	NN	O	O

Mutation	NN	O	O
of	NN	O	O
the	NN	O	O
core	NN	O	O
ETS	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
from	NN	O	O
-GGAA-	NN	O	B-DNA
to	NN	O	I-DNA
-GGAT-	NN	O	I-DNA
prevents	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
ETS-like	NN	O	B-protein
factors	NN	O	I-protein
with	NN	O	O
the	NN	O	O
exception	NN	O	O
of	NN	O	O
ETS1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
GM-CSF	NN	O	B-DNA
promoter	NN	O	I-DNA
,	NN	O	O
modified	NN	O	O
in	NN	O	O
this	NN	O	O
way	NN	O	O
to	NN	O	O
be	NN	O	O
ETS1	NN	O	B-protein
specific	NN	O	O
,	NN	O	O
is	NN	O	O
fully	NN	O	O
responsive	NN	O	O
to	NN	O	O
PMA/ionomycin	NN	O	O
induction	NN	O	O
,	NN	O	O
in	NN	O	O
addition	NN	O	O
to	NN	O	O
ETS1	NN	O	B-protein
transactivation	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
ionomycin	NN	O	O
.	NN	O	O

Together	NN	O	O
these	NN	O	O
data	NN	O	O
suggest	NN	O	O
that	NN	O	O
ETS1	NN	O	B-protein
may	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
mediating	NN	O	O
the	NN	O	O
increased	NN	O	O
GM-CSF	NN	O	B-protein
production	NN	O	O
associated	NN	O	O
with	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Quantitation	NN	O	O
of	NN	O	O
beta	NN	O	B-RNA
1	NN	O	I-RNA
triiodothyronine	NN	O	I-RNA
receptor	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
human	NN	O	O
tissues	NN	O	O
by	NN	O	O
competitive	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

Thyroid	NN	O	O
hormones	NN	O	O
act	NN	O	O
by	NN	O	O
binding	NN	O	O
to	NN	O	O
nuclear	NN	O	B-protein
receptor	NN	O	I-protein
proteins	NN	O	I-protein
,	NN	O	O
the	NN	O	O
thyroid	NN	O	B-protein
hormone	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	I-protein
TR	NN	O	I-protein
)	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	I-protein
beta	NN	O	I-protein
.	NN	O	O

Data	NN	O	O
from	NN	O	O
cell	NN	O	O
culture	NN	O	O
and	NN	O	O
animal	NN	O	O
studies	NN	O	O
indicate	NN	O	O
that	NN	O	O
TR	NN	O	B-protein
expression	NN	O	O
may	NN	O	O
be	NN	O	O
regulated	NN	O	O
to	NN	O	O
modulate	NN	O	O
target	NN	O	O
organ	NN	O	O
responsiveness	NN	O	O
to	NN	O	O
thyroid	NN	O	O
hormone	NN	O	O
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
whether	NN	O	O
such	NN	O	O
adaptive	NN	O	O
changes	NN	O	O
in	NN	O	O
TR	NN	O	B-protein
expression	NN	O	O
occur	NN	O	O
in	NN	O	O
humans	NN	O	O
,	NN	O	O
we	NN	O	O
determined	NN	O	O
the	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
of	NN	O	O
the	NN	O	O
hTR	NN	O	B-protein
beta	NN	O	I-protein
1	NN	O	I-protein
in	NN	O	O
various	NN	O	O
thyroid	NN	O	O
states	NN	O	O
.	NN	O	O

Patients	NN	O	O
with	NN	O	O
overt	NN	O	O
hypo	NN	O	O
-or	NN	O	O
hyperthyroidism	NN	O	O
were	NN	O	O
enrolled	NN	O	O
in	NN	O	O
the	NN	O	O
study	NN	O	O
.	NN	O	O

Total	NN	O	O
RNA	NN	O	O
was	NN	O	O
isolated	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
and	NN	O	O
hTR	NN	O	B-RNA
beta	NN	O	I-RNA
1	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
determined	NN	O	O
by	NN	O	O
quantitative	NN	O	O
competitive	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
PCR	NN	O	O
.	NN	O	O

For	NN	O	O
comparison	NN	O	O
,	NN	O	O
hTR	NN	O	B-RNA
beta	NN	O	I-RNA
1	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
were	NN	O	O
determined	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
and	NN	O	O
normal	NN	O	O
thyroid	NN	O	O
tissue	NN	O	O
of	NN	O	O
euthyroid	NN	O	O
patients	NN	O	O
.	NN	O	O

Human	NN	O	B-RNA
TR	NN	O	I-RNA
beta	NN	O	I-RNA
1	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
in	NN	O	O
lymphocytes	NN	O	B-cell_type
were	NN	O	O
1.8	NN	O	O
+/-	NN	O	O
0.4	NN	O	O
,	NN	O	O
1.9	NN	O	O
+/-	NN	O	O
0.5	NN	O	O
,	NN	O	O
1.1	NN	O	O
+/-	NN	O	O
0.4	NN	O	O
10	NN	O	O
(	NN	O	O
-18	NN	O	O
)	NN	O	O
mol/microgram	NN	O	O
RNA	NN	O	O
in	NN	O	O
hypo-	NN	O	O
,	NN	O	O
eu	NN	O	O
-and	NN	O	O
hyperthyroid	NN	O	O
patients	NN	O	O
,	NN	O	O
respectively	NN	O	O
,	NN	O	O
corresponding	NN	O	O
to	NN	O	O
an	NN	O	O
estimated	NN	O	O
0.5	NN	O	O
-2	NN	O	O
molecules	NN	O	O
per	NN	O	O
cell	NN	O	O
.	NN	O	O

Although	NN	O	O
the	NN	O	O
mean	NN	O	O
hTR	NN	O	B-RNA
beta	NN	O	I-RNA
1	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
were	NN	O	O
40	NN	O	O
%	NN	O	O
lower	NN	O	O
in	NN	O	O
hyperthyroid	NN	O	O
than	NN	O	O
in	NN	O	O
euthyroid	NN	O	O
subjects	NN	O	O
,	NN	O	O
this	NN	O	O
difference	NN	O	O
did	NN	O	O
not	NN	O	O
reach	NN	O	O
statistical	NN	O	O
significance	NN	O	O
.	NN	O	O

Similar	NN	O	O
levels	NN	O	O
of	NN	O	O
hTR	NN	O	B-RNA
beta	NN	O	I-RNA
1	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
were	NN	O	O
detected	NN	O	O
in	NN	O	O
thyroid	NN	O	O
gland	NN	O	O
from	NN	O	O
euthyroid	NN	O	O
patients	NN	O	O
.	NN	O	O

In	NN	O	O
summary	NN	O	O
,	NN	O	O
we	NN	O	O
developed	NN	O	O
an	NN	O	O
assay	NN	O	O
for	NN	O	O
the	NN	O	O
quantitative	NN	O	O
determination	NN	O	O
of	NN	O	O
hTR	NN	O	B-RNA
beta	NN	O	I-RNA
1	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
in	NN	O	O
small	NN	O	O
human	NN	O	O
tissue	NN	O	O
samples	NN	O	O
,	NN	O	O
containing	NN	O	O
as	NN	O	O
little	NN	O	O
as	NN	O	O
50	NN	O	O
ng	NN	O	O
of	NN	O	O
total	NN	O	O
RNA	NN	O	O
.	NN	O	O

Absolute	NN	O	O
hTR	NN	O	B-RNA
beta	NN	O	I-RNA
1	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
are	NN	O	O
very	NN	O	O
low	NN	O	O
with	NN	O	O
an	NN	O	O
estimated	NN	O	O
one	NN	O	O
molecule	NN	O	O
of	NN	O	O
mRNA	NN	O	B-RNA
being	NN	O	O
present	NN	O	O
in	NN	O	O
a	NN	O	O
mononuclear	NN	O	B-cell_type
blood	NN	O	I-cell_type
cell	NN	O	I-cell_type
or	NN	O	O
thyrocyte	NN	O	B-cell_type
.	NN	O	O

No	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
hTR	NN	O	B-protein
beta	NN	O	I-protein
1	NN	O	I-protein
was	NN	O	O
seen	NN	O	O
in	NN	O	O
hypothyroid	NN	O	O
relative	NN	O	O
to	NN	O	O
euthyroid	NN	O	O
patients	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
there	NN	O	O
is	NN	O	O
a	NN	O	O
non-significant	NN	O	O
trend	NN	O	O
towards	NN	O	O
a	NN	O	O
down-regulation	NN	O	O
of	NN	O	O
hTR	NN	O	B-RNA
beta	NN	O	I-RNA
1	NN	O	I-RNA
mRNA	NN	O	I-RNA
levels	NN	O	O
in	NN	O	O
hyperthyroid	NN	O	O
patients	NN	O	O
.	NN	O	O

-DOCSTART-	O

Multiple	NN	O	O
proteins	NN	O	O
interact	NN	O	O
with	NN	O	O
the	NN	O	O
nuclear	NN	O	B-DNA
inhibitory	NN	O	I-DNA
protein	NN	O	I-DNA
repressor	NN	O	I-DNA
element	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
interleukin-3	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

T	NN	O	O
cell	NN	O	O
expression	NN	O	O
of	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
is	NN	O	O
directed	NN	O	O
by	NN	O	O
positive	NN	O	B-DNA
and	NN	O	I-DNA
negative	NN	O	I-DNA
cis-acting	NN	O	I-DNA
DNA	NN	O	I-DNA
elements	NN	O	I-DNA
clustered	NN	O	O
within	NN	O	O
300	NN	O	O
base	NN	O	O
pairs	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

A	NN	O	O
strong	NN	O	O
repressor	NN	O	B-protein
element	NN	O	I-protein
,	NN	O	O
termed	NN	O	O
nuclear	NN	O	B-protein
inhibitory	NN	O	I-protein
protein	NN	O	I-protein
(	NN	O	O
NIP	NN	O	B-protein
)	NN	O	O
,	NN	O	O
was	NN	O	O
previously	NN	O	O
mapped	NN	O	O
to	NN	O	O
a	NN	O	O
segment	NN	O	O
of	NN	O	O
the	NN	O	O
IL-3	NN	O	B-DNA
promoter	NN	O	I-DNA
between	NN	O	O
nucleotides	NN	O	B-DNA
-271	NN	O	I-DNA
and	NN	O	I-DNA
-250	NN	O	I-DNA
.	NN	O	O

Functional	NN	O	O
characterization	NN	O	O
of	NN	O	O
this	NN	O	O
element	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
it	NN	O	O
can	NN	O	O
mediate	NN	O	O
repression	NN	O	O
when	NN	O	O
linked	NN	O	O
in	NN	O	O
cis	NN	O	O
to	NN	O	O
a	NN	O	O
heterologous	NN	O	B-DNA
promoter	NN	O	I-DNA
.	NN	O	O

DNA	NN	O	O
binding	NN	O	O
experiments	NN	O	O
were	NN	O	O
carried	NN	O	O
out	NN	O	O
to	NN	O	O
characterize	NN	O	O
the	NN	O	O
repressor	NN	O	O
activity	NN	O	O
.	NN	O	O

Using	NN	O	O
varying	NN	O	O
conditions	NN	O	O
,	NN	O	O
three	NN	O	O
distinct	NN	O	O
complexes	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
interact	NN	O	O
specifically	NN	O	O
with	NN	O	O
the	NN	O	O
NIP	NN	O	B-DNA
region	NN	O	I-DNA
,	NN	O	O
although	NN	O	O
only	NN	O	O
one	NN	O	O
correlates	NN	O	O
with	NN	O	O
repressor	NN	O	O
activity	NN	O	O
.	NN	O	O

Complex	NN	O	B-protein
1	NN	O	I-protein
results	NN	O	O
from	NN	O	O
binding	NN	O	O
of	NN	O	O
a	NN	O	O
ubiquitous	NN	O	O
polypeptide	NN	O	O
that	NN	O	O
recognizes	NN	O	O
the	NN	O	O
3	NN	O	B-DNA
'	NN	O	I-DNA
portion	NN	O	I-DNA
of	NN	O	O
this	NN	O	O
sequence	NN	O	O
and	NN	O	O
is	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
repression	NN	O	O
.	NN	O	O

Complex	NN	O	B-protein
2	NN	O	I-protein
corresponds	NN	O	O
to	NN	O	O
binding	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	O
upstream	NN	O	B-protein
stimulatory	NN	O	I-protein
factor	NN	O	I-protein
)	NN	O	O
to	NN	O	O
an	NN	O	O
E-box	NN	O	B-DNA
motif	NN	O	I-DNA
in	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
the	NN	O	O
NIP	NN	O	B-DNA
region	NN	O	I-DNA
.	NN	O	O

DNA	NN	O	O
binding	NN	O	O
specificity	NN	O	O
of	NN	O	O
complex	NN	O	O
3	NN	O	O
overlaps	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
upstream	NN	O	B-protein
stimulatory	NN	O	I-protein
factor	NN	O	I-protein
but	NN	O	O
is	NN	O	O
clearly	NN	O	O
distinct	NN	O	O
.	NN	O	O

To	NN	O	O
determine	NN	O	O
which	NN	O	O
of	NN	O	O
the	NN	O	O
latter	NN	O	O
two	NN	O	O
complexes	NN	O	O
represents	NN	O	O
NIP	NN	O	B-protein
activity	NN	O	O
,	NN	O	O
we	NN	O	O
incorporated	NN	O	O
small	NN	O	O
alterations	NN	O	O
into	NN	O	O
the	NN	O	O
NIP	NN	O	B-DNA
site	NN	O	I-DNA
of	NN	O	O
an	NN	O	O
IL-3	NN	O	B-DNA
promoter-linked	NN	O	I-DNA
reporter	NN	O	I-DNA
construct	NN	O	I-DNA
and	NN	O	O
examined	NN	O	O
their	NN	O	O
effects	NN	O	O
on	NN	O	O
NIP	NN	O	B-protein
-mediated	NN	O	O
repression	NN	O	O
.	NN	O	O

Functional	NN	O	O
specificity	NN	O	O
for	NN	O	O
repression	NN	O	O
matches	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
specificity	NN	O	O
of	NN	O	O
complex	NN	O	B-protein
3	NN	O	I-protein
;	NN	O	O
both	NN	O	O
repressor	NN	O	O
activity	NN	O	O
and	NN	O	O
complex	NN	O	B-protein
3	NN	O	I-protein
binding	NN	O	O
require	NN	O	O
the	NN	O	O
consensus	NN	O	B-DNA
sequence	NN	O	I-DNA
CTCACNTNC	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
hematopoietic	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
PU.1	NN	O	B-protein
is	NN	O	O
downregulated	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
multiple	NN	O	I-cell_line
myeloma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

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
belonging	NN	O	O
to	NN	O	O
the	NN	O	O
Ets-family	NN	O	B-protein
.	NN	O	O

It	NN	O	O
is	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
Spi-1	NN	O	B-DNA
oncogene	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
implicated	NN	O	O
in	NN	O	O
spleen	NN	O	O
focus-forming	NN	O	O
virus-induced	NN	O	O
murine	NN	O	O
erythroleukemias	NN	O	O
.	NN	O	O

PU.1	NN	O	B-protein
seems	NN	O	O
to	NN	O	O
be	NN	O	O
required	NN	O	O
for	NN	O	O
early	NN	O	O
development	NN	O	O
of	NN	O	O
multiple	NN	O	B-cell_type
hematopoietic	NN	O	I-cell_type
lineages	NN	O	I-cell_type
,	NN	O	O
but	NN	O	O
its	NN	O	O
expression	NN	O	O
in	NN	O	O
mature	NN	O	B-cell_type
cells	NN	O	I-cell_type
is	NN	O	O
preferentially	NN	O	O
observed	NN	O	O
in	NN	O	O
cells	NN	O	O
of	NN	O	O
the	NN	O	O
B-cell-and	NN	O	B-cell_type
monocyte/macrophage-differentiation	NN	O	I-cell_type
lineage	NN	O	I-cell_type
.	NN	O	O

It	NN	O	O
binds	NN	O	O
the	NN	O	O
so-called	NN	O	O
Pu	NN	O	B-DNA
box	NN	O	I-DNA
,	NN	O	O
an	NN	O	O
important	NN	O	O
tissue-specific	NN	O	B-DNA
regulatory	NN	O	I-DNA
DNA	NN	O	I-DNA
element	NN	O	I-DNA
present	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
genes	NN	O	O
expressed	NN	O	O
in	NN	O	O
these	NN	O	O
cell	NN	O	B-cell_type
lineages	NN	O	I-cell_type
.	NN	O	O

We	NN	O	O
have	NN	O	O
analyzed	NN	O	O
the	NN	O	O
expression	NN	O	O
and	NN	O	O
activity	NN	O	O
of	NN	O	O
PU.1	NN	O	B-protein
during	NN	O	O
human	NN	O	O
B-cell	NN	O	O
development	NN	O	O
using	NN	O	O
a	NN	O	O
panel	NN	O	O
of	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
maturation	NN	O	O
,	NN	O	O
from	NN	O	O
early	NN	O	B-cell_line
precursors	NN	O	I-cell_line
to	NN	O	O
differentiated	NN	O	B-cell_line
plasma	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

PU.1	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
and	NN	O	O
PU.1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
as	NN	O	O
measured	NN	O	O
by	NN	O	O
Northern	NN	O	O
blot	NN	O	O
analysis	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
were	NN	O	O
evident	NN	O	O
in	NN	O	O
cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
representing	NN	O	O
pro-B	NN	O	B-cell_line
,	NN	O	I-cell_line
pre-B	NN	O	I-cell_line
,	NN	O	I-cell_line
and	NN	O	I-cell_line
mature	NN	O	I-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

We	NN	O	O
could	NN	O	O
also	NN	O	O
show	NN	O	O
Pu	NN	O	B-DNA
box	NN	O	I-DNA
-dependent	NN	O	O
transactivation	NN	O	O
of	NN	O	O
a	NN	O	O
reporter	NN	O	O
gene	NN	O	O
in	NN	O	O
transient	NN	O	O
transfections	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

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
in	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
multiple	NN	O	B-cell_line
myeloma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
representing	NN	O	O
differentiated	NN	O	B-cell_type
,	NN	O	I-cell_type
plasma	NN	O	I-cell_type
cell-like	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
PU.1	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
,	NN	O	O
mRNA	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
Pu	NN	O	B-DNA
box	NN	O	I-DNA
-dependent	NN	O	O
transactivation	NN	O	O
were	NN	O	O
absent	NN	O	O
or	NN	O	O
detectable	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

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
which	NN	O	O
exemplify	NN	O	O
an	NN	O	O
intermediate	NN	O	O
stage	NN	O	O
of	NN	O	O
B-cell	NN	O	O
differentiation	NN	O	O
,	NN	O	O
a	NN	O	O
reduced	NN	O	O
expression	NN	O	O
and	NN	O	O
activity	NN	O	O
were	NN	O	O
observed	NN	O	O
.	NN	O	O

The	NN	O	O
findings	NN	O	O
in	NN	O	O
the	NN	O	O
human	NN	O	B-cell_line
multiple	NN	O	I-cell_line
myeloma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
represent	NN	O	O
the	NN	O	O
first	NN	O	O
examples	NN	O	O
of	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
downregulated	NN	O	O
PU.1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
apparently	NN	O	O
contradict	NN	O	O
observations	NN	O	O
in	NN	O	O
the	NN	O	O
murine	NN	O	O
system	NN	O	O
in	NN	O	O
which	NN	O	O
PU.1	NN	O	B-protein
is	NN	O	O
expressed	NN	O	O
and	NN	O	O
active	NN	O	O
in	NN	O	O
plasmacytoma	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

At	NN	O	O
present	NN	O	O
,	NN	O	O
it	NN	O	O
is	NN	O	O
unclear	NN	O	O
whether	NN	O	O
the	NN	O	O
lack	NN	O	O
of	NN	O	O
PU.1	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
multiple	NN	O	I-cell_line
myeloma	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
represents	NN	O	O
a	NN	O	O
malignancy-associated	NN	O	O
defect	NN	O	O
in	NN	O	O
these	NN	O	O
cells	NN	O	O
or	NN	O	O
exemplifies	NN	O	O
a	NN	O	O
normal	NN	O	O
developmental	NN	O	O
regulation	NN	O	O
in	NN	O	O
terminally	NN	O	B-cell_type
differentiated	NN	O	I-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	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
and	NN	O	O
E-selectin	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
and	NN	O	O
the	NN	O	O
T-cell	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
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
NFAT	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
T-cell-specific	NN	O	B-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
athat	NN	O	O
supported	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
and	NN	O	O
mediated	NN	O	O
the	NN	O	O
immunoregulatory	NN	O	B-protein
effects	NN	O	O
of	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
.	NN	O	O

As	NN	O	O
we	NN	O	O
observed	NN	O	O
that	NN	O	O
activated	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
also	NN	O	O
expressed	NN	O	O
NFAT	NN	O	B-protein
,	NN	O	O
we	NN	O	O
tested	NN	O	O
the	NN	O	O
antiinflammatory	NN	O	O
properties	NN	O	O
of	NN	O	O
CsA	NN	O	O
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Significantly	NN	O	O
,	NN	O	O
CsA	NN	O	O
completely	NN	O	O
suppressed	NN	O	O
the	NN	O	O
induction	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
inhibited	NN	O	O
the	NN	O	O
activity	NN	O	O
of	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
regulatory	NN	O	I-DNA
elements	NN	O	I-DNA
that	NN	O	O
use	NN	O	O
NFAT	NN	O	B-protein
by	NN	O	O
60	NN	O	O
%	NN	O	O
.	NN	O	O

CsA	NN	O	O
similarly	NN	O	O
mediated	NN	O	O
a	NN	O	O
reduction	NN	O	O
of	NN	O	O
up	NN	O	O
to	NN	O	O
65	NN	O	O
%	NN	O	O
in	NN	O	O
GM-CSF	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
protein	NN	O	O
expression	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

CsA	NN	O	O
also	NN	O	O
suppressed	NN	O	O
E-selectin	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	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
expression	NN	O	O
in	NN	O	O
endothelial	NN	O	O
cells	NN	O	O
,	NN	O	O
even	NN	O	O
though	NN	O	O
the	NN	O	O
E-selectin	NN	O	B-DNA
promoter	NN	O	I-DNA
is	NN	O	O
activated	NN	O	O
by	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
rather	NN	O	O
than	NN	O	O
NFAT	NN	O	B-protein
.	NN	O	O

Hence	NN	O	O
,	NN	O	O
induction	NN	O	O
of	NN	O	O
cell	NN	O	O
surface	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
leukocyte	NN	O	B-protein
adhesion	NN	O	I-protein
molecule	NN	O	I-protein
by	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
was	NN	O	O
reduced	NN	O	O
by	NN	O	O
40	NN	O	O
%	NN	O	O
in	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
CsA	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
was	NN	O	O
reflected	NN	O	O
by	NN	O	O
a	NN	O	O
29	NN	O	O
%	NN	O	O
decrease	NN	O	O
in	NN	O	O
neutrophil	NN	O	B-cell_type
adhesion	NN	O	O
.	NN	O	O

The	NN	O	O
effects	NN	O	O
of	NN	O	O
CsA	NN	O	O
on	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
also	NN	O	O
detected	NN	O	O
at	NN	O	O
the	NN	O	O
chromatin	NN	O	B-DNA
structure	NN	O	O
level	NN	O	O
,	NN	O	O
as	NN	O	O
DNasel	NN	O	B-DNA
hypersensitive	NN	O	I-DNA
sites	NN	O	I-DNA
within	NN	O	O
both	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-protein
enhancer	NN	O	O
and	NN	O	O
the	NN	O	O
E-selectin	NN	O	B-DNA
promoter	NN	O	I-DNA
were	NN	O	O
suppressed	NN	O	O
by	NN	O	O
CsA	NN	O	O
.	NN	O	O

This	NN	O	O
represents	NN	O	O
the	NN	O	O
first	NN	O	O
report	NN	O	O
of	NN	O	O
NFAT	NN	O	B-protein
in	NN	O	O
endothelial	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
suggests	NN	O	O
mechanisms	NN	O	O
by	NN	O	O
which	NN	O	O
CsA	NN	O	O
could	NN	O	O
function	NN	O	O
as	NN	O	O
an	NN	O	O
antiinflammatory	NN	O	O
agent	NN	O	O
.	NN	O	O

-DOCSTART-	O

Costimulation	NN	O	O
requirement	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
transcription	NN	O	O
factor	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

The	NN	O	O
transcriptional	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
requires	NN	O	O
T-cell	NN	O	B-cell_type
costimulation	NN	O	O
delivered	NN	O	O
by	NN	O	O
the	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
the	NN	O	O
auxiliary	NN	O	B-protein
receptor	NN	O	I-protein
CD28	NN	O	I-protein
.	NN	O	O

Several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
participate	NN	O	O
in	NN	O	O
IL-2	NN	O	B-protein
promoter	NN	O	O
activation	NN	O	O
,	NN	O	O
among	NN	O	O
which	NN	O	O
are	NN	O	O
AP-1-like	NN	O	B-protein
factors	NN	O	I-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
.	NN	O	O

Protein	NN	O	O
phosphorylation	NN	O	O
has	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
these	NN	O	O
two	NN	O	O
factors	NN	O	O
:	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
it	NN	O	O
induces	NN	O	O
the	NN	O	O
transactivating	NN	O	O
capacity	NN	O	O
of	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
protein	NN	O	I-protein
c-Jun	NN	O	B-protein
;	NN	O	O
and	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
it	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
the	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
,	NN	O	O
from	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
,	NN	O	O
allowing	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
latter	NN	O	O
into	NN	O	O
the	NN	O	O
nucleus	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
both	NN	O	O
phosphorylation	NN	O	O
processes	NN	O	O
require	NN	O	O
T-cell	NN	O	O
costimulation	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	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
kinetics	NN	O	O
of	NN	O	O
the	NN	O	O
two	NN	O	O
phosphorylation	NN	O	O
events	NN	O	O
are	NN	O	O
essentially	NN	O	O
similar	NN	O	O
.	NN	O	O

According	NN	O	O
to	NN	O	O
our	NN	O	O
results	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
the	NN	O	O
kinases	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
two	NN	O	O
processes	NN	O	O
are	NN	O	O
distinct	NN	O	O
entities	NN	O	O
.	NN	O	O

Whereas	NN	O	O
TPCK	NN	O	B-protein
inhibits	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
and	NN	O	O
,	NN	O	O
consequently	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
,	NN	O	O
it	NN	O	O
markedly	NN	O	O
enhances	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
JNK	NN	O	B-protein
,	NN	O	O
the	NN	O	O
MAP	NN	O	B-protein
kinase-related	NN	O	I-protein
kinase	NN	O	I-protein
that	NN	O	O
phosphorylates	NN	O	O
the	NN	O	O
transactivation	NN	O	B-protein
domain	NN	O	I-protein
of	NN	O	O
c-Jun	NN	O	B-protein
.	NN	O	O

We	NN	O	O
,	NN	O	O
therefore	NN	O	O
,	NN	O	O
propose	NN	O	O
the	NN	O	O
activation	NN	O	O
scheme	NN	O	O
presented	NN	O	O
in	NN	O	O
FIGURE	NN	O	O
3	NN	O	O
for	NN	O	O
T-cell	NN	O	O
costimulation	NN	O	O
.	NN	O	O

Costimulation	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
a	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
that	NN	O	O
leads	NN	O	O
to	NN	O	O
the	NN	O	O
simultaneous	NN	O	O
induction	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
,	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

Integration	NN	O	O
of	NN	O	O
the	NN	O	O
signals	NN	O	O
generated	NN	O	O
by	NN	O	O
TCR	NN	O	B-protein
and	NN	O	O
CD28	NN	O	B-protein
engagement	NN	O	O
occurs	NN	O	O
along	NN	O	O
this	NN	O	O
pathway	NN	O	O
,	NN	O	O
which	NN	O	O
then	NN	O	O
bifurcates	NN	O	O
to	NN	O	O
induce	NN	O	O
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
phosphorylation	NN	O	O
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
on	NN	O	O
the	NN	O	O
one	NN	O	O
hand	NN	O	O
,	NN	O	O
and	NN	O	O
JNK	NN	O	B-protein
activation	NN	O	O
and	NN	O	O
c-Jun	NN	O	B-protein
phosphorylation	NN	O	O
on	NN	O	O
the	NN	O	O
other	NN	O	O
.	NN	O	O

We	NN	O	O
are	NN	O	O
currently	NN	O	O
engaged	NN	O	O
in	NN	O	O
defining	NN	O	O
where	NN	O	O
the	NN	O	O
two	NN	O	O
signals	NN	O	O
integrate	NN	O	O
along	NN	O	O
the	NN	O	O
AP-1	NN	O	B-protein
/	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
pathway	NN	O	O
.	NN	O	O

-DOCSTART-	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
and	NN	O	O
cytokine	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
NF-kappa	NN	O	B-protein
B/Rel	NN	O	I-protein
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

CD4+	NN	O	B-cell_line
macrophages	NN	O	I-cell_line
in	NN	O	O
tissues	NN	O	O
such	NN	O	O
as	NN	O	O
lung	NN	O	O
,	NN	O	O
skin	NN	O	O
,	NN	O	O
and	NN	O	O
lymph	NN	O	O
nodes	NN	O	O
,	NN	O	O
promyelocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
in	NN	O	O
bone	NN	O	O
marrow	NN	O	O
,	NN	O	O
and	NN	O	O
peripheral	NN	O	B-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
serve	NN	O	O
as	NN	O	O
important	NN	O	O
targets	NN	O	O
and	NN	O	O
reservoirs	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
replication	NN	O	O
.	NN	O	O

HIV-1-infected	NN	O	O
myeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
are	NN	O	O
often	NN	O	O
diminished	NN	O	O
in	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
participate	NN	O	O
in	NN	O	O
chemotaxis	NN	O	O
,	NN	O	O
phagocytosis	NN	O	O
,	NN	O	O
and	NN	O	O
intracellular	NN	O	O
killing	NN	O	O
.	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
can	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
surface	NN	O	B-protein
receptors	NN	O	I-protein
associated	NN	O	O
with	NN	O	O
cellular	NN	O	O
activation	NN	O	O
and/or	NN	O	O
differentiation	NN	O	O
that	NN	O	O
increase	NN	O	O
the	NN	O	O
responsiveness	NN	O	O
of	NN	O	O
these	NN	O	O
cells	NN	O	O
to	NN	O	O
cytokines	NN	O	B-protein
secreted	NN	O	O
by	NN	O	O
neighboring	NN	O	O
cells	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
to	NN	O	O
bacteria	NN	O	O
or	NN	O	O
other	NN	O	O
pathogens	NN	O	O
.	NN	O	O

Enhancement	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
is	NN	O	O
related	NN	O	O
in	NN	O	O
part	NN	O	O
to	NN	O	O
increased	NN	O	O
DNA-binding	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
such	NN	O	O
as	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
binds	NN	O	O
to	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
region	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
and	NN	O	O
contributes	NN	O	O
to	NN	O	O
the	NN	O	O
inducibility	NN	O	O
of	NN	O	O
HIV-1	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
multiple	NN	O	O
activating	NN	O	O
agents	NN	O	O
.	NN	O	O

Phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
the	NN	O	O
cytoplasmic	NN	O	B-protein
inhibitor	NN	O	I-protein
I	NN	O	B-protein
kappa	NN	O	I-protein
B	NN	O	I-protein
alpha	NN	O	O
are	NN	O	O
crucial	NN	O	O
regulatory	NN	O	O
events	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
DNA-binding	NN	O	O
activity	NN	O	O
.	NN	O	O

Both	NN	O	O
N-	NN	O	O
and	NN	O	O
C-terminal	NN	O	O
residues	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
are	NN	O	O
required	NN	O	O
for	NN	O	O
inducer-mediated	NN	O	O
degradation	NN	O	O
.	NN	O	O

Chronic	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
leads	NN	O	O
to	NN	O	O
constitutive	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
DNA-binding	NN	O	O
activity	NN	O	O
and	NN	O	O
provides	NN	O	O
an	NN	O	O
intranuclear	NN	O	O
environment	NN	O	O
capable	NN	O	O
of	NN	O	O
perpetuating	NN	O	O
HIV-1	NN	O	O
replication	NN	O	O
.	NN	O	O

Increased	NN	O	O
intracellular	NN	O	O
stores	NN	O	O
of	NN	O	O
latent	NN	O	B-protein
NF-kappa	NN	O	I-protein
B	NN	O	I-protein
may	NN	O	O
also	NN	O	O
result	NN	O	O
in	NN	O	O
rapid	NN	O	O
inducibility	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
-dependent	NN	O	O
cytokine	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

In	NN	O	O
response	NN	O	O
to	NN	O	O
secondary	NN	O	O
pathogenic	NN	O	O
infections	NN	O	O
or	NN	O	O
antigenic	NN	O	O
challenge	NN	O	O
,	NN	O	O
cytokine	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
is	NN	O	O
rapidly	NN	O	O
induced	NN	O	O
,	NN	O	O
enhanced	NN	O	O
,	NN	O	O
and	NN	O	O
sustained	NN	O	O
over	NN	O	O
prolonged	NN	O	O
periods	NN	O	O
in	NN	O	O
HIV-1-infected	NN	O	B-cell_line
myeloid	NN	O	I-cell_line
cells	NN	O	I-cell_line
compared	NN	O	O
with	NN	O	O
uninfected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Elevated	NN	O	O
levels	NN	O	O
of	NN	O	O
several	NN	O	O
inflammatory	NN	O	B-protein
cytokines	NN	O	I-protein
have	NN	O	O
been	NN	O	O
detected	NN	O	O
in	NN	O	O
the	NN	O	O
sera	NN	O	O
of	NN	O	O
HIV-1-infected	NN	O	O
individuals	NN	O	O
.	NN	O	O

Secretion	NN	O	O
of	NN	O	O
myeloid	NN	O	B-protein
cell-derived	NN	O	I-protein
cytokines	NN	O	I-protein
may	NN	O	O
both	NN	O	O
increase	NN	O	O
virus	NN	O	O
production	NN	O	O
and	NN	O	O
contribute	NN	O	O
to	NN	O	O
AIDS-associated	NN	O	O
disorders	NN	O	O
.	NN	O	O

-DOCSTART-	O

Steroid	NN	O	O
mediated	NN	O	O
lysis	NN	O	O
of	NN	O	O
lymphoblasts	NN	O	B-cell_type
requires	NN	O	O
the	NN	O	O
DNA	NN	O	O
binding	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	O
hormone	NN	O	O
receptor	NN	O	O
.	NN	O	O

Glucocorticoids	NN	O	O
kill	NN	O	O
certain	NN	O	O
types	NN	O	O
of	NN	O	O
lymphoblasts	NN	O	B-cell_type
,	NN	O	O
but	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
are	NN	O	O
unknown	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
clear	NN	O	O
that	NN	O	O
sufficient	NN	O	O
numbers	NN	O	O
of	NN	O	O
functional	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
are	NN	O	O
required	NN	O	O
to	NN	O	O
mediate	NN	O	O
lysis	NN	O	O
,	NN	O	O
but	NN	O	O
whether	NN	O	O
they	NN	O	O
do	NN	O	O
so	NN	O	O
through	NN	O	O
the	NN	O	O
classical	NN	O	O
model	NN	O	O
of	NN	O	O
steroid	NN	O	O
hormone	NN	O	O
activation	NN	O	O
and	NN	O	O
modulation	NN	O	O
of	NN	O	O
gene	NN	O	O
expression	NN	O	O
has	NN	O	O
not	NN	O	O
been	NN	O	O
established	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
asked	NN	O	O
which	NN	O	O
region	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
of	NN	O	O
the	NN	O	O
steroid	NN	O	B-protein
receptor	NN	O	I-protein
are	NN	O	O
important	NN	O	O
for	NN	O	O
mediating	NN	O	O
lysis	NN	O	O
in	NN	O	O
leukemic	NN	O	B-cell_type
T	NN	O	I-cell_type
lymphoblasts	NN	O	I-cell_type
.	NN	O	O

CEM-ICR	NN	O	B-cell_line
27	NN	O	I-cell_line
leukemic	NN	O	I-cell_line
lymphoblasts	NN	O	I-cell_line
,	NN	O	O
a	NN	O	O
clone	NN	O	O
of	NN	O	O
CEM	NN	O	B-cell_line
cells	NN	O	I-cell_line
which	NN	O	O
lack	NN	O	O
functional	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
and	NN	O	O
therefore	NN	O	O
are	NN	O	O
neither	NN	O	O
lysed	NN	O	O
by	NN	O	O
dexamethasone	NN	O	O
nor	NN	O	O
capable	NN	O	O
of	NN	O	O
showing	NN	O	O
glutamine	NN	O	B-protein
synthetase	NN	O	I-protein
induction	NN	O	O
,	NN	O	O
were	NN	O	O
provided	NN	O	O
with	NN	O	O
steroid	NN	O	B-protein
receptors	NN	O	I-protein
by	NN	O	O
DNA	NN	O	O
transfections	NN	O	O
of	NN	O	O
various	NN	O	O
receptor	NN	O	O
gene	NN	O	O
constructs	NN	O	O
.	NN	O	O

We	NN	O	O
measured	NN	O	O
steroid	NN	O	O
mediated	NN	O	O
lysis	NN	O	O
,	NN	O	O
receptor	NN	O	O
number	NN	O	O
and	NN	O	O
induction	NN	O	O
of	NN	O	O
glutamine	NN	O	B-protein
synthetase	NN	O	I-protein
in	NN	O	O
the	NN	O	O
transfected	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
results	NN	O	O
provide	NN	O	O
evidence	NN	O	O
that	NN	O	O
the	NN	O	O
lysis	NN	O	O
mechanism	NN	O	O
in	NN	O	O
the	NN	O	O
ICR27	NN	O	B-cell_line
lymphoblasts	NN	O	I-cell_line
is	NN	O	O
restored	NN	O	O
when	NN	O	O
functional	NN	O	O
receptor	NN	O	O
number	NN	O	O
is	NN	O	O
restored	NN	O	O
.	NN	O	O

The	NN	O	O
DNA	NN	O	B-protein
binding	NN	O	I-protein
region	NN	O	I-protein
specifying	NN	O	O
high	NN	O	O
affinity	NN	O	O
for	NN	O	O
GRE	NN	O	B-DNA
sites	NN	O	I-DNA
is	NN	O	O
required	NN	O	O
.	NN	O	O

Lysis	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
any	NN	O	O
steroid	NN	O	O
that	NN	O	O
allows	NN	O	O
for	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
containing	NN	O	O
such	NN	O	O
a	NN	O	O
region	NN	O	O
.	NN	O	O

Our	NN	O	O
data	NN	O	O
support	NN	O	O
the	NN	O	O
view	NN	O	O
that	NN	O	O
steroid-mediated	NN	O	O
cell	NN	O	O
death	NN	O	O
occurs	NN	O	O
by	NN	O	O
a	NN	O	O
process	NN	O	O
requiring	NN	O	O
direct	NN	O	O
interaction	NN	O	O
of	NN	O	O
steroid-receptor	NN	O	B-protein
complexes	NN	O	I-protein
with	NN	O	O
the	NN	O	O
genome	NN	O	O
.	NN	O	O

-DOCSTART-	O

Functional	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
homolog	NN	O	I-protein
of	NN	O	O
the	NN	O	O
B	NN	O	B-protein
cell-specific	NN	O	I-protein
coactivator	NN	O	I-protein
BOB.1/OBF.1	NN	O	B-protein
.	NN	O	O

B	NN	O	O
cell-specific	NN	O	O
transcriptional	NN	O	O
promoter	NN	O	O
activity	NN	O	O
mediated	NN	O	O
by	NN	O	O
the	NN	O	O
octamer	NN	O	B-DNA
motif	NN	O	I-DNA
requires	NN	O	O
the	NN	O	O
Oct1	NN	O	B-protein
or	NN	O	O
Oct2	NN	O	B-protein
protein	NN	O	I-protein
and	NN	O	O
additional	NN	O	O
B	NN	O	B-protein
cell-restricted	NN	O	I-protein
cofactors	NN	O	I-protein
.	NN	O	O

One	NN	O	O
such	NN	O	O
cofactor	NN	O	B-protein
,	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
,	NN	O	O
was	NN	O	O
recently	NN	O	O
isolated	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

Here	NN	O	O
,	NN	O	O
we	NN	O	O
describe	NN	O	O
the	NN	O	O
isolation	NN	O	O
and	NN	O	O
detailed	NN	O	O
characterization	NN	O	O
of	NN	O	O
the	NN	O	O
murine	NN	O	B-protein
homolog	NN	O	I-protein
.	NN	O	O

Full-length	NN	O	O
cDNAs	NN	O	B-DNA
and	NN	O	O
genomic	NN	O	B-DNA
clones	NN	O	I-DNA
were	NN	O	O
isolated	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
gene	NN	O	O
structure	NN	O	O
was	NN	O	O
determined.	NN	O	O
Comparison	NN	O	O
of	NN	O	O
the	NN	O	O
deduced	NN	O	O
amino	NN	O	O
acids	NN	O	O
shows	NN	O	O
88	NN	O	O
%	NN	O	O
sequence	NN	O	O
identity	NN	O	O
between	NN	O	O
mouse	NN	O	B-protein
and	NN	O	I-protein
human	NN	O	I-protein
BOB.1/OBF.1	NN	O	I-protein
.	NN	O	O

The	NN	O	O
NH2-terminal	NN	O	B-protein
126	NN	O	I-protein
amino	NN	O	I-protein
acids	NN	O	I-protein
of	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
are	NN	O	O
both	NN	O	O
essential	NN	O	O
and	NN	O	O
sufficient	NN	O	O
for	NN	O	O
interaction	NN	O	O
with	NN	O	O
the	NN	O	O
POU	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
either	NN	O	O
Oct1	NN	O	B-protein
or	NN	O	O
Oct2	NN	O	B-protein
.	NN	O	O

This	NN	O	O
protein-protein	NN	O	O
interaction	NN	O	O
does	NN	O	O
not	NN	O	O
require	NN	O	O
the	NN	O	O
simultaneous	NN	O	O
binding	NN	O	O
of	NN	O	O
Oct	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
DNA	NN	O	O
,	NN	O	O
and	NN	O	O
high	NN	O	O
resolution	NN	O	O
footprinting	NN	O	O
of	NN	O	O
the	NN	O	O
Oct	NN	O	B-DNA
-DNA	NN	O	O
interaction	NN	O	O
reveals	NN	O	O
that	NN	O	O
binding	NN	O	O
of	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
to	NN	O	O
Oct1	NN	O	B-protein
or	NN	O	O
Oct2	NN	O	B-protein
does	NN	O	O
not	NN	O	O
alter	NN	O	O
the	NN	O	O
interaction	NN	O	O
with	NN	O	O
DNA	NN	O	O
.	NN	O	O

BOB.1/OBF.1	NN	O	B-protein
can	NN	O	O
efficiently	NN	O	O
activate	NN	O	O
octamer-dependent	NN	O	O
promoters	NN	O	O
in	NN	O	O
fibroblasts	NN	O	B-cell_type
;	NN	O	O
however	NN	O	O
,	NN	O	O
it	NN	O	O
fails	NN	O	O
to	NN	O	O
stimulate	NN	O	O
octamer-dependent	NN	O	B-DNA
enhancer	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

Fusion	NN	O	O
of	NN	O	O
subdomains	NN	O	O
of	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
with	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
reveals	NN	O	O
that	NN	O	O
both	NN	O	O
NH2-	NN	O	B-protein
and	NN	O	I-protein
COOH-terminal	NN	O	I-protein
domains	NN	O	I-protein
of	NN	O	O
BOB.1/OBF.1	NN	O	B-protein
contribute	NN	O	O
to	NN	O	O
full	NN	O	O
transactivation	NN	O	O
function	NN	O	O
,	NN	O	O
the	NN	O	O
COOH-terminal	NN	O	B-protein
domain	NN	O	I-protein
is	NN	O	O
more	NN	O	O
efficient	NN	O	O
in	NN	O	O
this	NN	O	O
transactivation	NN	O	O
assay	NN	O	O
.	NN	O	O

Consistent	NN	O	O
with	NN	O	O
the	NN	O	O
failure	NN	O	O
of	NN	O	O
full-length	NN	O	B-protein
BOB.1/OBF.1	NN	O	I-protein
to	NN	O	O
stimulate	NN	O	O
octamer-dependent	NN	O	B-DNA
enhancer	NN	O	I-DNA
elements	NN	O	I-DNA
in	NN	O	O
non	NN	O	B-cell_type
B	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
GAL4	NN	O	B-protein
fusions	NN	O	I-protein
likewise	NN	O	O
only	NN	O	O
stimulate	NN	O	O
from	NN	O	O
a	NN	O	O
promoter-proximal	NN	O	O
position	NN	O	O
.	NN	O	O

-DOCSTART-	O

Anti-immunoglobulin	NN	O	B-protein
M	NN	O	I-protein
activates	NN	O	O
nuclear	NN	O	B-protein
calcium/calmodulin-dependent	NN	O	I-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
II	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

We	NN	O	O
and	NN	O	O
others	NN	O	O
have	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
Ets-1	NN	O	B-protein
,	NN	O	O
is	NN	O	O
phosphorylated	NN	O	O
in	NN	O	O
a	NN	O	O
calcium-dependent	NN	O	O
manner	NN	O	O
after	NN	O	O
ligation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	O
(	NN	O	O
Ig	NN	O	O
)	NN	O	O
M	NN	O	O
on	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

As	NN	O	O
this	NN	O	O
phosphorylation	NN	O	O
was	NN	O	O
independent	NN	O	O
of	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
we	NN	O	O
tested	NN	O	O
whether	NN	O	O
a	NN	O	O
calcium/calmodulin-dependent	NN	O	B-protein
protein	NN	O	I-protein
kinase	NN	O	I-protein
(	NN	O	O
CaM	NN	O	B-protein
kinase	NN	O	I-protein
)	NN	O	O
might	NN	O	O
phosphorylate	NN	O	O
the	NN	O	O
Ets-1	NN	O	B-protein
protein	NN	O	I-protein
after	NN	O	O
elevation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
free	NN	O	O
calcium	NN	O	O
concentrations	NN	O	O
.	NN	O	O

The	NN	O	O
dephosphorylated	NN	O	O
form	NN	O	O
of	NN	O	O
Ets-1	NN	O	B-protein
has	NN	O	O
been	NN	O	O
shown	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
chromatin	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
the	NN	O	O
operative	NN	O	B-protein
kinase	NN	O	I-protein
should	NN	O	O
be	NN	O	O
detectable	NN	O	O
in	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

We	NN	O	O
prepared	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
from	NN	O	O
two	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
in	NN	O	O
which	NN	O	O
increased	NN	O	O
intracellular	NN	O	O
free	NN	O	O
calcium	NN	O	O
levels	NN	O	O
correlated	NN	O	O
with	NN	O	O
increased	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
Ets-1	NN	O	B-protein
protein	NN	O	O
.	NN	O	O

Activity	NN	O	O
of	NN	O	O
the	NN	O	O
CaM	NN	O	B-protein
kinases	NN	O	I-protein
was	NN	O	O
determined	NN	O	O
using	NN	O	O
a	NN	O	O
synthetic	NN	O	O
peptide	NN	O	O
substrate	NN	O	O
both	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
inhibitor	NN	O	O
specific	NN	O	O
for	NN	O	O
the	NN	O	O
CaM	NN	O	B-protein
kinase	NN	O	I-protein
family	NN	O	I-protein
,	NN	O	O
KN-62	NN	O	O
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
cells	NN	O	O
with	NN	O	O
anti-IgM	NN	O	B-protein
led	NN	O	O
to	NN	O	O
increased	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
kinase	NN	O	I-protein
that	NN	O	O
could	NN	O	O
phosphorylate	NN	O	O
the	NN	O	O
peptide	NN	O	O
,	NN	O	O
and	NN	O	O
this	NN	O	O
activity	NN	O	O
was	NN	O	O
reduced	NN	O	O
by	NN	O	O
10	NN	O	O
microM	NN	O	O
KN-62	NN	O	O
.	NN	O	O

Kinase	NN	O	O
activity	NN	O	O
was	NN	O	O
reduced	NN	O	O
in	NN	O	O
lysates	NN	O	O
preadsorbed	NN	O	O
using	NN	O	O
an	NN	O	O
antibody	NN	O	B-protein
specific	NN	O	O
for	NN	O	O
CaM	NN	O	B-protein
kinase	NN	O	I-protein
II	NN	O	I-protein
.	NN	O	O

Two-dimensional	NN	O	O
phosphopeptide	NN	O	O
maps	NN	O	O
of	NN	O	O
the	NN	O	O
Ets-1	NN	O	B-protein
protein	NN	O	O
from	NN	O	O
cells	NN	O	O
incubated	NN	O	O
with	NN	O	O
ionomycin	NN	O	O
or	NN	O	O
anti-IgM	NN	O	B-protein
contained	NN	O	O
two	NN	O	O
unique	NN	O	O
phosphopeptides	NN	O	O
that	NN	O	O
were	NN	O	O
absent	NN	O	O
in	NN	O	O
untreated	NN	O	O
cells	NN	O	O
.	NN	O	O

Incubation	NN	O	O
of	NN	O	O
isolated	NN	O	B-protein
Ets-1	NN	O	I-protein
protein	NN	O	I-protein
with	NN	O	O
purified	NN	O	O
CaM	NN	O	B-protein
kinase	NN	O	I-protein
II	NN	O	I-protein
produced	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
peptides	NN	O	O
that	NN	O	O
migrated	NN	O	O
identically	NN	O	O
to	NN	O	O
those	NN	O	O
found	NN	O	O
in	NN	O	O
cells	NN	O	O
incubated	NN	O	O
with	NN	O	O
either	NN	O	O
anti-IgM	NN	O	B-protein
or	NN	O	O
ionomycin	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
of	NN	O	O
signal	NN	O	O
transduction	NN	O	O
by	NN	O	O
the	NN	O	O
antigen	NN	O	B-protein
receptor	NN	O	I-protein
on	NN	O	O
B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
which	NN	O	O
increased	NN	O	O
intracellular	NN	O	O
free	NN	O	O
calcium	NN	O	O
can	NN	O	O
rapidly	NN	O	O
activate	NN	O	O
nuclear	NN	O	B-protein
CaM	NN	O	I-protein
kinase	NN	O	I-protein
II	NN	O	I-protein
,	NN	O	O
potentially	NN	O	O
resulting	NN	O	O
in	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
regulation	NN	O	O
of	NN	O	O
DNA-binding	NN	O	B-protein
proteins	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
activation	NN	O	O
and	NN	O	O
repression	NN	O	O
,	NN	O	O
two	NN	O	O
properties	NN	O	O
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
contains	NN	O	O
two	NN	O	O
transcriptional	NN	O	B-protein
activation	NN	O	I-protein
domains	NN	O	I-protein
which	NN	O	O
are	NN	O	O
located	NN	O	O
within	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
and	NN	O	O
C-terminal	NN	O	B-protein
regions	NN	O	I-protein
.	NN	O	O

To	NN	O	O
study	NN	O	O
their	NN	O	O
differential	NN	O	O
activation	NN	O	O
properties	NN	O	O
,	NN	O	O
we	NN	O	O
linked	NN	O	O
the	NN	O	O
isolated	NN	O	O
effector	NN	O	B-protein
domains	NN	O	I-protein
to	NN	O	O
the	NN	O	O
GAL4	NN	O	B-protein
DNA-binding	NN	O	I-protein
domain	NN	O	I-protein
.	NN	O	O

We	NN	O	O
have	NN	O	O
shown	NN	O	O
that	NN	O	O
both	NN	O	O
activating	NN	O	O
regions	NN	O	O
of	NN	O	O
Oct-2a	NN	O	B-protein
,	NN	O	O
isolated	NN	O	O
from	NN	O	O
their	NN	O	O
natural	NN	O	O
context	NN	O	O
,	NN	O	O
can	NN	O	O
activate	NN	O	O
transcription	NN	O	O
as	NN	O	O
promoter	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
to	NN	O	O
the	NN	O	O
C-terminus	NN	O	B-protein
,	NN	O	O
activation	NN	O	O
by	NN	O	O
the	NN	O	O
N-terminal	NN	O	B-protein
domain	NN	O	I-protein
is	NN	O	O
dependent	NN	O	O
on	NN	O	O
a	NN	O	O
yet	NN	O	O
unidentified	NN	O	O
factor	NN	O	O
(	NN	O	O
s	NN	O	O
)	NN	O	O
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
.	NN	O	O

The	NN	O	O
results	NN	O	O
obtained	NN	O	O
by	NN	O	O
duplication	NN	O	O
of	NN	O	O
activation	NN	O	B-protein
domains	NN	O	I-protein
or	NN	O	O
their	NN	O	O
mixed	NN	O	O
combination	NN	O	O
suggest	NN	O	O
that	NN	O	O
the	NN	O	O
domains	NN	O	O
are	NN	O	O
functionally	NN	O	O
independent	NN	O	O
.	NN	O	O

However	NN	O	O
,	NN	O	O
activation	NN	O	O
from	NN	O	O
a	NN	O	O
remote	NN	O	O
position	NN	O	O
could	NN	O	O
only	NN	O	O
be	NN	O	O
achieved	NN	O	O
with	NN	O	O
the	NN	O	O
C-terminus	NN	O	B-protein
of	NN	O	O
Oct-2a	NN	O	B-protein
in	NN	O	O
B	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
lymphoid	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
higher	NN	O	O
activation	NN	O	O
levels	NN	O	O
were	NN	O	O
observed	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
distinct	NN	O	O
B-cell-specific	NN	O	B-protein
cofactors	NN	O	I-protein
in	NN	O	O
concert	NN	O	O
with	NN	O	O
the	NN	O	O
effector	NN	O	B-protein
domains	NN	O	I-protein
of	NN	O	O
Oct-2a	NN	O	B-protein
might	NN	O	O
be	NN	O	O
involved	NN	O	O
in	NN	O	O
mediating	NN	O	O
transcription	NN	O	O
from	NN	O	O
proximal	NN	O	O
and	NN	O	O
remote	NN	O	O
positions	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
we	NN	O	O
identified	NN	O	O
a	NN	O	O
repression	NN	O	O
domain	NN	O	O
at	NN	O	O
the	NN	O	O
N-terminus	NN	O	O
of	NN	O	O
Oct-2a	NN	O	B-protein
.	NN	O	O

When	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
transcriptional	NN	O	O
stimulation	NN	O	O
was	NN	O	O
inhibited	NN	O	O
efficiently	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
underscore	NN	O	O
the	NN	O	O
modular	NN	O	O
structure	NN	O	O
of	NN	O	O
Oct-2a	NN	O	B-protein
with	NN	O	O
separable	NN	O	O
domains	NN	O	O
for	NN	O	O
activation	NN	O	O
and	NN	O	O
repression	NN	O	O
and	NN	O	O
suggest	NN	O	O
that	NN	O	O
Oct-2a	NN	O	B-protein
might	NN	O	O
have	NN	O	O
complex	NN	O	O
regulatory	NN	O	O
functions	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

-DOCSTART-	O

Nonopsonic	NN	O	O
phagocytosis	NN	O	O
of	NN	O	O
Pseudomonas	NN	O	O
aeruginosa	NN	O	O
by	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
polymorphonuclear	NN	O	B-cell_type
leukocytes	NN	O	I-cell_type
requires	NN	O	O
the	NN	O	O
presence	NN	O	O
of	NN	O	O
the	NN	O	O
bacterial	NN	O	O
flagellum	NN	O	O
.	NN	O	O

Whereas	NN	O	O
the	NN	O	O
mechanism	NN	O	O
of	NN	O	O
nonopsonic	NN	O	O
phagocytosis	NN	O	O
of	NN	O	O
Pseudomonas	NN	O	O
aeruginosa	NN	O	O
has	NN	O	O
been	NN	O	O
described	NN	O	O
,	NN	O	O
the	NN	O	O
bacterial	NN	O	O
ligands	NN	O	O
required	NN	O	O
are	NN	O	O
poorly	NN	O	O
understood	NN	O	O
.	NN	O	O

To	NN	O	O
identify	NN	O	O
the	NN	O	O
requisite	NN	O	O
bacterial	NN	O	O
ligands	NN	O	O
,	NN	O	O
studies	NN	O	O
with	NN	O	O
isogenic	NN	O	O
mutants	NN	O	O
of	NN	O	O
P.	NN	O	O
aeruginosa	NN	O	O
PAK	NN	O	O
lacking	NN	O	O
pili	NN	O	O
,	NN	O	O
flagella	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
RpoN	NN	O	B-protein
sigma	NN	O	I-protein
factor	NN	O	I-protein
were	NN	O	O
undertaken	NN	O	O
.	NN	O	O

The	NN	O	O
RpoN	NN	O	O
mutant	NN	O	O
,	NN	O	O
lacking	NN	O	O
pili	NN	O	O
,	NN	O	O
flagella	NN	O	O
,	NN	O	O
and	NN	O	O
nonpilus	NN	O	B-protein
adhesins	NN	O	I-protein
,	NN	O	O
bound	NN	O	O
poorly	NN	O	O
and	NN	O	O
was	NN	O	O
resistant	NN	O	O
to	NN	O	O
ingestion	NN	O	O
by	NN	O	O
both	NN	O	O
macrophages	NN	O	B-cell_type
and	NN	O	O
neutrophils	NN	O	B-cell_type
.	NN	O	O

Pili	NN	O	O
were	NN	O	O
not	NN	O	O
absolutely	NN	O	O
required	NN	O	O
for	NN	O	O
binding	NN	O	O
or	NN	O	O
phagocytosis	NN	O	O
of	NN	O	O
P.	NN	O	O
aeruginosa	NN	O	O
.	NN	O	O

The	NN	O	O
presence	NN	O	O
of	NN	O	O
a	NN	O	O
flagellum	NN	O	O
was	NN	O	O
not	NN	O	O
required	NN	O	O
for	NN	O	O
binding	NN	O	O
of	NN	O	O
P.	NN	O	O
aeruginosa	NN	O	O
to	NN	O	O
macrophages	NN	O	B-cell_type
but	NN	O	O
was	NN	O	O
critical	NN	O	O
for	NN	O	O
the	NN	O	O
subsequent	NN	O	O
internalization	NN	O	O
of	NN	O	O
the	NN	O	O
bacterium	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
this	NN	O	O
factor	NN	O	O
or	NN	O	O
a	NN	O	O
surface	NN	O	O
ligand	NN	O	O
associated	NN	O	O
with	NN	O	O
its	NN	O	O
assembly	NN	O	O
was	NN	O	O
responsible	NN	O	O
for	NN	O	O
stimulation	NN	O	O
of	NN	O	O
nonopsonic	NN	O	O
phagocytosis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Identification	NN	O	O
of	NN	O	O
essential	NN	O	O
GATA	NN	O	B-DNA
and	NN	O	O
Ets	NN	O	B-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
promoter	NN	O	O
of	NN	O	O
the	NN	O	O
platelet	NN	O	B-DNA
glycoprotein	NN	O	I-DNA
Ib	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Platelet	NN	O	B-protein
glycoprotein	NN	O	I-protein
(	NN	O	O
GP	NN	O	B-protein
)	NN	O	O
Ib-IX-V	NN	O	B-protein
is	NN	O	O
a	NN	O	O
multisubunit	NN	O	B-protein
adhesion	NN	O	I-protein
receptor	NN	O	I-protein
that	NN	O	O
supports	NN	O	O
platelet	NN	O	O
attachment	NN	O	O
to	NN	O	O
thrombogenic	NN	O	O
surfaces	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

The	NN	O	O
congenital	NN	O	O
absence	NN	O	O
of	NN	O	O
the	NN	O	O
receptor	NN	O	O
results	NN	O	O
in	NN	O	O
a	NN	O	O
bleeding	NN	O	O
disorder	NN	O	O
associated	NN	O	O
with	NN	O	O
``	NN	O	B-cell_type
giant	NN	O	I-cell_type
''	NN	O	I-cell_type
platelets	NN	O	I-cell_type
,	NN	O	O
a	NN	O	O
condition	NN	O	O
linking	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
complex	NN	O	O
to	NN	O	O
platelet	NN	O	O
morphogenesis	NN	O	O
.	NN	O	O

To	NN	O	O
understand	NN	O	O
better	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
GP	NN	O	B-protein
Ib-IX-V	NN	O	I-protein
complex	NN	O	I-protein
,	NN	O	O
studies	NN	O	O
were	NN	O	O
undertaken	NN	O	O
to	NN	O	O
define	NN	O	O
the	NN	O	O
essential	NN	O	O
genetic	NN	O	B-DNA
elements	NN	O	I-DNA
supporting	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
alpha-subunit	NN	O	B-protein
of	NN	O	O
the	NN	O	O
complex	NN	O	O
(	NN	O	O
GP	NN	O	B-protein
Ib	NN	O	I-protein
alpha	NN	O	I-protein
)	NN	O	O
.	NN	O	O

GP	NN	O	B-DNA
Ib	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
was	NN	O	O
evaluated	NN	O	O
by	NN	O	O
transfection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
erythroleukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
with	NN	O	O
reporter	NN	O	O
plasmids	NN	O	O
coding	NN	O	O
for	NN	O	O
the	NN	O	O
enzyme	NN	O	B-protein
,	NN	O	O
luciferase	NN	O	B-protein
.	NN	O	O

Studies	NN	O	O
were	NN	O	O
initiated	NN	O	O
with	NN	O	O
a	NN	O	O
fragment	NN	O	O
extending	NN	O	O
2	NN	O	O
,	NN	O	O
738	NN	O	O
nucleotides	NN	O	O
5	NN	O	O
'	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
and	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
identification	NN	O	O
of	NN	O	O
253	NN	O	O
nucleotides	NN	O	O
retaining	NN	O	O
full	NN	O	O
promoter	NN	O	O
activity	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
erythroleukemia	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

In	NN	O	O
cells	NN	O	O
of	NN	O	O
nonhematopoietic	NN	O	B-cell_type
lineage	NN	O	I-cell_type
,	NN	O	O
human	NN	O	B-cell_type
endothelial	NN	O	I-cell_type
and	NN	O	O
HeLa	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
the	NN	O	O
GP	NN	O	B-DNA
Ib	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
was	NN	O	O
no	NN	O	O
greater	NN	O	O
than	NN	O	O
background	NN	O	O
levels	NN	O	O
obtained	NN	O	O
with	NN	O	O
promoterless	NN	O	B-DNA
constructs	NN	O	I-DNA
.	NN	O	O

Gel	NN	O	O
shift	NN	O	O
assays	NN	O	O
and	NN	O	O
site-directed	NN	O	O
mutagenesis	NN	O	O
studies	NN	O	O
defined	NN	O	O
essential	NN	O	O
GATA	NN	O	B-DNA
and	NN	O	O
Ets	NN	O	B-DNA
binding	NN	O	I-DNA
motifs	NN	O	I-DNA
93	NN	O	B-DNA
and	NN	O	I-DNA
150	NN	O	I-DNA
nucleotides	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
,	NN	O	O
a	NN	O	O
finding	NN	O	O
which	NN	O	O
further	NN	O	O
substantiates	NN	O	O
these	NN	O	O
elements	NN	O	O
as	NN	O	O
important	NN	O	O
determinants	NN	O	O
of	NN	O	O
megakaryocytic	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

The	NN	O	O
results	NN	O	O
define	NN	O	O
essential	NN	O	O
cis-acting	NN	O	B-DNA
elements	NN	O	I-DNA
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
GP	NN	O	B-protein
Ib	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
provide	NN	O	O
insights	NN	O	O
into	NN	O	O
molecular	NN	O	O
events	NN	O	O
coinciding	NN	O	O
with	NN	O	O
the	NN	O	O
release	NN	O	O
of	NN	O	O
normal	NN	O	B-cell_type
platelets	NN	O	I-cell_type
into	NN	O	O
the	NN	O	O
bloodstream	NN	O	O
.	NN	O	O

-DOCSTART-	O

CD30	NN	O	B-protein
ligation	NN	O	O
induces	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
human	NN	O	B-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

CD30	NN	O	B-protein
is	NN	O	O
a	NN	O	O
recently	NN	O	O
described	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/nerve	NN	O	I-protein
growth	NN	O	I-protein
factor	NN	O	I-protein
receptor	NN	O	I-protein
superfamily	NN	O	I-protein
.	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
following	NN	O	O
incubation	NN	O	O
of	NN	O	O
L540	NN	O	B-cell_line
cells	NN	O	I-cell_line
(	NN	O	O
Hodgkin	NN	O	B-cell_line
's	NN	O	I-cell_line
disease-derived	NN	O	I-cell_line
,	NN	O	I-cell_line
T	NN	O	I-cell_line
cell-like	NN	O	I-cell_line
,	NN	O	I-cell_line
CD30+	NN	O	I-cell_line
cells	NN	O	I-cell_line
)	NN	O	O
with	NN	O	O
the	NN	O	O
agonistic	NN	O	B-protein
anti-CD30	NN	O	I-protein
monoclonal	NN	O	I-protein
antibodies	NN	O	I-protein
(	NN	O	O
mAb	NN	O	B-protein
)	NN	O	O
M44	NN	O	B-protein
and	NN	O	O
M67	NN	O	B-protein
,	NN	O	O
two	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
DNA	NN	O	O
binding	NN	O	O
activities	NN	O	O
were	NN	O	O
induced	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
retardation	NN	O	O
assays	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
the	NN	O	O
mAb	NN	O	B-protein
towards	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activation	NN	O	O
was	NN	O	O
rapid	NN	O	O
,	NN	O	O
as	NN	O	O
it	NN	O	O
occurred	NN	O	O
within	NN	O	O
20	NN	O	O
min	NN	O	O
,	NN	O	O
and	NN	O	O
was	NN	O	O
sustained	NN	O	O
for	NN	O	O
up	NN	O	O
to	NN	O	O
6	NN	O	O
h	NN	O	O
.	NN	O	O

By	NN	O	O
comparison	NN	O	O
,	NN	O	O
an	NN	O	O
isotype-matched	NN	O	B-protein
antibody	NN	O	I-protein
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
.	NN	O	O

Moreover	NN	O	O
,	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
T	NN	O	I-cell_line
helper	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
functionally	NN	O	O
characterized	NN	O	O
as	NN	O	O
being	NN	O	O
of	NN	O	O
the	NN	O	O
type	NN	O	O
0	NN	O	O
,	NN	O	O
type	NN	O	O
1	NN	O	O
and	NN	O	O
type	NN	O	O
2	NN	O	O
(	NN	O	O
28	NN	O	O
%	NN	O	O
,	NN	O	O
<	NN	O	O
1	NN	O	O
%	NN	O	O
und	NN	O	O
93	NN	O	O
%	NN	O	O
CD30+	NN	O	O
,	NN	O	O
respectively	NN	O	O
)	NN	O	O
,	NN	O	O
the	NN	O	O
extent	NN	O	O
of	NN	O	O
CD30	NN	O	B-protein
-mediated	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
proportion	NN	O	O
of	NN	O	O
CD30+	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
all	NN	O	O
cell	NN	O	O
lines	NN	O	O
investigated	NN	O	O
,	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
complexes	NN	O	I-protein
induced	NN	O	O
following	NN	O	O
CD30	NN	O	B-protein
engagement	NN	O	O
were	NN	O	O
shown	NN	O	O
to	NN	O	O
contain	NN	O	O
p50	NN	O	B-protein
NF-kappa	NN	O	I-protein
B1	NN	O	I-protein
,	NN	O	O
p65	NN	O	B-protein
RelA	NN	O	I-protein
,	NN	O	O
and	NN	O	O
possibly	NN	O	O
other	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
.	NN	O	O

Collectively	NN	O	O
,	NN	O	O
our	NN	O	O
results	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
nuclear	NN	O	O
translocation	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
rank	NN	O	O
among	NN	O	O
the	NN	O	O
short-term	NN	O	O
cellular	NN	O	O
responses	NN	O	O
elicited	NN	O	O
following	NN	O	O
CD30	NN	O	B-protein
ligation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Pathogenesis	NN	O	O
of	NN	O	O
atherosclerosis	NN	O	O
.	NN	O	O

The	NN	O	O
earliest	NN	O	O
lesion	NN	O	O
in	NN	O	O
the	NN	O	O
development	NN	O	O
of	NN	O	O
an	NN	O	O
atherosclerotic	NN	O	O
plaque	NN	O	O
is	NN	O	O
the	NN	O	O
fatty	NN	O	O
streak	NN	O	O
.	NN	O	O

This	NN	O	O
chronic	NN	O	O
inflammatory	NN	O	O
reaction	NN	O	O
results	NN	O	O
from	NN	O	O
a	NN	O	O
sequence	NN	O	O
of	NN	O	O
events	NN	O	O
that	NN	O	O
begins	NN	O	O
with	NN	O	O
the	NN	O	O
trapping	NN	O	O
of	NN	O	O
low	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
(	NN	O	O
LDL	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
subendothelial	NN	O	O
space	NN	O	O
of	NN	O	O
the	NN	O	O
artery	NN	O	O
wall	NN	O	O
.	NN	O	O

The	NN	O	O
trapped	NN	O	O
LDL	NN	O	O
is	NN	O	O
seeded	NN	O	O
with	NN	O	O
oxidative	NN	O	O
species	NN	O	O
released	NN	O	O
by	NN	O	O
the	NN	O	O
overlying	NN	O	O
endothelium	NN	O	O
,	NN	O	O
and	NN	O	O
lipid	NN	O	O
oxidation	NN	O	O
is	NN	O	O
initiated	NN	O	O
within	NN	O	O
the	NN	O	O
LDL	NN	O	O
particle.	NN	O	O
Some	NN	O	O
of	NN	O	O
the	NN	O	O
lipids	NN	O	O
that	NN	O	O
result	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
NFkB-like	NN	O	B-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
that	NN	O	O
cause	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
genes	NN	O	O
whose	NN	O	O
protein	NN	O	B-protein
products	NN	O	I-protein
mediate	NN	O	O
monocyte	NN	O	O
binding	NN	O	O
,	NN	O	O
monocyte	NN	O	B-cell_type
chemotaxis	NN	O	O
into	NN	O	O
the	NN	O	O
subendothelial	NN	O	O
space	NN	O	O
,	NN	O	O
and	NN	O	O
conversion	NN	O	O
into	NN	O	O
macrophages	NN	O	B-cell_type
.	NN	O	O

At	NN	O	O
least	NN	O	O
1	NN	O	O
major	NN	O	B-DNA
gene	NN	O	I-DNA
modulates	NN	O	O
the	NN	O	O
oxidation	NN	O	O
of	NN	O	O
LDL	NN	O	O
lipids	NN	O	O
and/or	NN	O	O
the	NN	O	O
biologic	NN	O	O
response	NN	O	O
to	NN	O	O
these	NN	O	O
lipids	NN	O	O
.	NN	O	O

The	NN	O	O
inverse	NN	O	O
relation	NN	O	O
between	NN	O	O
high	NN	O	O
density	NN	O	O
lipoprotein	NN	O	O
(	NN	O	O
HDL	NN	O	O
)	NN	O	O
and	NN	O	O
atherosclerotic	NN	O	O
events	NN	O	O
may	NN	O	O
in	NN	O	O
part	NN	O	O
be	NN	O	O
due	NN	O	O
to	NN	O	O
enzymes	NN	O	B-protein
associated	NN	O	O
with	NN	O	O
HDL	NN	O	O
that	NN	O	O
destroy	NN	O	O
the	NN	O	O
biologically	NN	O	O
active	NN	O	O
lipids	NN	O	O
generated	NN	O	O
in	NN	O	O
LDL	NN	O	O
.	NN	O	O

-DOCSTART-	O

Estrogen	NN	O	B-protein
and	NN	O	O
progesterone	NN	O	B-protein
receptors	NN	O	I-protein
in	NN	O	O
vernal	NN	O	O
keratoconjunctivitis	NN	O	O
.	NN	O	O

PURPOSE	NN	O	O
:	NN	O	O
Sex-related	NN	O	O
influences	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
vernal	NN	O	O
keratoconjunctivitis	NN	O	O
(	NN	O	O
VKC	NN	O	O
)	NN	O	O
,	NN	O	O
an	NN	O	O
allergic	NN	O	O
eosinophilic	NN	O	O
disease	NN	O	O
.	NN	O	O

METHODS	NN	O	O
:	NN	O	O
The	NN	O	O
authors	NN	O	O
evaluated	NN	O	O
tarsal	NN	O	O
and	NN	O	O
bulbar	NN	O	O
conjunctival	NN	O	O
biopsies	NN	O	O
from	NN	O	O
seven	NN	O	O
patients	NN	O	O
with	NN	O	O
severe	NN	O	O
and	NN	O	O
symptomatic	NN	O	O
VKC	NN	O	O
for	NN	O	O
the	NN	O	O
presence	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
by	NN	O	O
using	NN	O	O
monoclonal	NN	O	B-protein
antibodies	NN	O	I-protein
with	NN	O	O
a	NN	O	O
peroxidase-antiperoxidase	NN	O	O
technique	NN	O	O
.	NN	O	O

RESULTS	NN	O	O
:	NN	O	O
Both	NN	O	O
the	NN	O	O
epithelium	NN	O	O
and	NN	O	O
subepithelium	NN	O	O
of	NN	O	O
the	NN	O	O
tarsal	NN	O	O
and	NN	O	O
bulbar	NN	O	O
conjunctiva	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
VKC	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
those	NN	O	O
of	NN	O	O
four	NN	O	O
nonatopic	NN	O	O
control	NN	O	O
subjects	NN	O	O
,	NN	O	O
showed	NN	O	O
intense	NN	O	O
positive	NN	O	O
staining	NN	O	O
for	NN	O	O
estrogen	NN	O	O
and	NN	O	O
progesterone	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Immunofluorescence	NN	O	O
colocalization	NN	O	O
of	NN	O	O
both	NN	O	O
estrogen	NN	O	B-protein
and	NN	O	O
progesterone	NN	O	B-protein
receptors	NN	O	I-protein
with	NN	O	O
eosinophil	NN	O	B-protein
cationic	NN	O	I-protein
protein	NN	O	I-protein
showed	NN	O	O
that	NN	O	O
approximately	NN	O	O
70	NN	O	O
%	NN	O	O
of	NN	O	O
positive	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
eosinophils	NN	O	B-cell_type
.	NN	O	O

CONCLUSIONS	NN	O	O
:	NN	O	O
Sexual	NN	O	O
hormones	NN	O	O
,	NN	O	O
through	NN	O	O
their	NN	O	O
receptors	NN	O	B-protein
,	NN	O	O
may	NN	O	O
influence	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
eosinophils	NN	O	B-cell_type
in	NN	O	O
patients	NN	O	O
with	NN	O	O
VKC	NN	O	O
.	NN	O	O

-DOCSTART-	O

CIITA	NN	O	B-protein
activates	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
in	NN	O	O
mouse	NN	O	O
T	NN	O	O
cells	NN	O	O
.	NN	O	O

It	NN	O	O
has	NN	O	O
long	NN	O	O
been	NN	O	O
a	NN	O	O
puzzle	NN	O	O
that	NN	O	O
MHC	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
are	NN	O	O
expressed	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
after	NN	O	O
activation	NN	O	O
but	NN	O	O
not	NN	O	O
in	NN	O	O
mouse	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
;	NN	O	O
this	NN	O	O
expression	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
the	NN	O	O
cell	NN	O	O
mediated	NN	O	O
immune	NN	O	O
response	NN	O	O
.	NN	O	O

Recently	NN	O	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
has	NN	O	O
been	NN	O	O
reported	NN	O	O
to	NN	O	O
be	NN	O	O
a	NN	O	O
major	NN	O	B-protein
regulatory	NN	O	I-protein
factor	NN	O	I-protein
for	NN	O	O
both	NN	O	O
the	NN	O	O
constitutive	NN	O	O
and	NN	O	O
IFN	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

Here	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
human	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
expressing	NN	O	O
MHC	NN	O	O
class	NN	O	O
II	NN	O	O
have	NN	O	O
CIITA	NN	O	B-protein
transcripts	NN	O	O
while	NN	O	O
MHC	NN	O	O
class	NN	O	O
II-negative	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
mouse	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
.	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
in	NN	O	O
mouse	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
be	NN	O	O
reconstituted	NN	O	O
upon	NN	O	O
transfection	NN	O	O
with	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
CIITA	NN	O	I-DNA
cDNA	NN	O	I-DNA
.	NN	O	O

These	NN	O	O
data	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
CIITA	NN	O	B-protein
explains	NN	O	O
the	NN	O	O
expression	NN	O	O
or	NN	O	O
lack	NN	O	O
of	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
in	NN	O	O
human	NN	O	B-cell_type
and	NN	O	O
mouse	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
respectively	NN	O	O
.	NN	O	O

-DOCSTART-	O

Anti-Ro	NN	O	B-protein
(	NN	O	I-protein
SSA	NN	O	I-protein
)	NN	O	I-protein
autoantibodies	NN	O	I-protein
are	NN	O	O
associated	NN	O	O
with	NN	O	O
T	NN	O	B-DNA
cell	NN	O	I-DNA
receptor	NN	O	I-DNA
beta	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
patients	NN	O	O
.	NN	O	O

Several	NN	O	O
of	NN	O	O
the	NN	O	O
heterogeneous	NN	O	O
clinical	NN	O	O
manifestations	NN	O	O
of	NN	O	O
systemic	NN	O	O
lupus	NN	O	O
erythematosus	NN	O	O
have	NN	O	O
been	NN	O	O
associated	NN	O	O
with	NN	O	O
specific	NN	O	O
autoantibodies	NN	O	B-protein
.	NN	O	O

Associations	NN	O	O
between	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
autoantibodies	NN	O	B-protein
to	NN	O	O
the	NN	O	O
ribonucleoproteins	NN	O	B-protein
Ro	NN	O	B-protein
(	NN	O	I-protein
SSA	NN	O	I-protein
)	NN	O	I-protein
and	NN	O	O
La	NN	O	B-protein
(	NN	O	I-protein
SSB	NN	O	I-protein
)	NN	O	I-protein
have	NN	O	O
been	NN	O	O
reported	NN	O	O
in	NN	O	O
these	NN	O	O
patients	NN	O	O
.	NN	O	O

Because	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
molecules	NN	O	I-protein
present	NN	O	O
antigen	NN	O	O
to	NN	O	O
T	NN	O	B-protein
cell	NN	O	I-protein
receptors	NN	O	I-protein
(	NN	O	O
TCRs	NN	O	B-protein
)	NN	O	O
,	NN	O	O
we	NN	O	O
have	NN	O	O
searched	NN	O	O
for	NN	O	O
a	NN	O	O
TCR	NN	O	B-DNA
gene	NN	O	I-DNA
associated	NN	O	O
with	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
anti-Ro	NN	O	B-protein
(	NN	O	I-protein
SSA	NN	O	I-protein
)	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

A	NN	O	O
pair	NN	O	O
of	NN	O	O
restriction	NN	O	B-DNA
fragment	NN	O	I-DNA
length	NN	O	I-DNA
polymorphisms	NN	O	I-DNA
(	NN	O	O
RFLPs	NN	O	B-DNA
)	NN	O	O
,	NN	O	O
one	NN	O	O
of	NN	O	O
which	NN	O	O
hybridizes	NN	O	O
to	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
constant	NN	O	I-DNA
region	NN	O	I-DNA
C	NN	O	I-DNA
beta	NN	O	I-DNA
1	NN	O	I-DNA
and	NN	O	O
the	NN	O	O
other	NN	O	O
to	NN	O	O
the	NN	O	O
C	NN	O	B-DNA
beta	NN	O	I-DNA
2	NN	O	I-DNA
gene	NN	O	I-DNA
,	NN	O	O
has	NN	O	O
been	NN	O	O
identified	NN	O	O
,	NN	O	O
suggesting	NN	O	O
these	NN	O	O
may	NN	O	O
be	NN	O	O
genotypic	NN	O	O
markers	NN	O	O
for	NN	O	O
an	NN	O	O
extended	NN	O	O
region	NN	O	O
of	NN	O	O
the	NN	O	O
TCR	NN	O	B-DNA
beta	NN	O	I-DNA
locus	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
RFLP	NN	O	B-DNA
pair	NN	O	I-DNA
occurs	NN	O	O
in	NN	O	O
76	NN	O	O
%	NN	O	O
of	NN	O	O
patients	NN	O	O
with	NN	O	O
Ro	NN	O	B-protein
(	NN	O	I-protein
SSA	NN	O	I-protein
)	NN	O	I-protein
precipitins	NN	O	O
,	NN	O	O
84	NN	O	O
%	NN	O	O
of	NN	O	O
anti-	NN	O	O
Ro	NN	O	B-protein
(	NN	O	I-protein
SSA	NN	O	I-protein
)	NN	O	I-protein
-positive	NN	O	O
patients	NN	O	O
lacking	NN	O	O
La	NN	O	B-protein
(	NN	O	I-protein
SSB	NN	O	I-protein
)	NN	O	I-protein
precipitins	NN	O	B-protein
,	NN	O	O
but	NN	O	O
only	NN	O	O
41	NN	O	O
%	NN	O	O
of	NN	O	O
the	NN	O	O
patients	NN	O	O
lacking	NN	O	O
both	NN	O	O
precipitins	NN	O	B-protein
(	NN	O	O
P	NN	O	O
=	NN	O	O
0.0004	NN	O	O
)	NN	O	O
.	NN	O	O

This	NN	O	O
disproportionate	NN	O	O
occurrence	NN	O	O
in	NN	O	O
a	NN	O	O
subset	NN	O	O
of	NN	O	O
lupus	NN	O	O
patients	NN	O	O
indicates	NN	O	O
that	NN	O	O
these	NN	O	O
RFLPs	NN	O	B-DNA
are	NN	O	O
not	NN	O	O
disease	NN	O	O
susceptibility	NN	O	O
markers	NN	O	O
,	NN	O	O
but	NN	O	O
rather	NN	O	O
are	NN	O	O
important	NN	O	O
markers	NN	O	O
for	NN	O	O
TCR	NN	O	B-DNA
genes	NN	O	I-DNA
whose	NN	O	O
products	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
the	NN	O	O
production	NN	O	O
of	NN	O	O
anti-Ro	NN	O	B-protein
(	NN	O	I-protein
SSA	NN	O	I-protein
)	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

The	NN	O	O
majority	NN	O	O
of	NN	O	O
patients	NN	O	O
who	NN	O	O
have	NN	O	O
these	NN	O	O
RFLPs	NN	O	B-DNA
and	NN	O	O
HLA	NN	O	B-protein
class	NN	O	I-protein
II	NN	O	I-protein
antigens	NN	O	I-protein
previously	NN	O	O
associated	NN	O	O
with	NN	O	O
the	NN	O	O
anti-	NN	O	O
Ro	NN	O	B-protein
(	NN	O	I-protein
SSA	NN	O	I-protein
)	NN	O	I-protein
response	NN	O	O
make	NN	O	O
this	NN	O	O
antibody	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
interactions	NN	O	O
between	NN	O	O
products	NN	O	O
of	NN	O	O
these	NN	O	O
loci	NN	O	B-DNA
occur	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
Ro	NN	O	B-protein
(	NN	O	I-protein
SSA	NN	O	I-protein
)	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

Inhibition	NN	O	O
of	NN	O	O
NF-AT	NN	O	B-protein
-dependent	NN	O	O
transcription	NN	O	O
by	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
differential	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
helper	NN	O	I-cell_type
cell	NN	O	I-cell_type
subsets	NN	O	I-cell_type
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
individual	NN	O	O
CD4+	NN	O	B-cell_line
T	NN	O	I-cell_line
cells	NN	O	I-cell_line
results	NN	O	O
in	NN	O	O
differential	NN	O	O
lymphokine	NN	O	B-protein
expression	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
is	NN	O	O
preferentially	NN	O	O
produced	NN	O	O
by	NN	O	O
T	NN	O	B-cell_line
helper	NN	O	I-cell_line
type	NN	O	I-cell_line
1	NN	O	I-cell_line
(	NN	O	I-cell_line
TH1	NN	O	I-cell_line
)	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
are	NN	O	O
involved	NN	O	O
in	NN	O	O
cell-mediated	NN	O	O
immune	NN	O	O
responses	NN	O	O
,	NN	O	O
whereas	NN	O	O
IL-4	NN	O	B-protein
is	NN	O	O
synthesized	NN	O	O
by	NN	O	O
TH2	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
are	NN	O	O
essential	NN	O	O
for	NN	O	O
humoral	NN	O	O
immunity	NN	O	O
.	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
-dependent	NN	O	I-protein
factor	NN	O	I-protein
NF-ATp	NN	O	B-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
inducible	NN	O	O
transcription	NN	O	O
of	NN	O	O
both	NN	O	O
these	NN	O	O
lymphokine	NN	O	B-DNA
genes	NN	O	I-DNA
.	NN	O	O

However	NN	O	O
,	NN	O	O
while	NN	O	O
IL2	NN	O	B-protein
expression	NN	O	O
requires	NN	O	O
the	NN	O	O
contribution	NN	O	O
of	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-	NN	O	O
and	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
-dependent	NN	O	O
signals	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
human	NN	O	B-protein
IL4	NN	O	I-protein
transcription	NN	O	O
through	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
pathway	NN	O	O
is	NN	O	O
diminished	NN	O	O
by	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
stimulation	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
phenomenon	NN	O	O
is	NN	O	O
due	NN	O	O
to	NN	O	O
mutually	NN	O	O
exclusive	NN	O	O
binding	NN	O	O
of	NN	O	O
NF-ATp	NN	O	B-protein
and	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
to	NN	O	O
the	NN	O	O
P	NN	O	B-DNA
sequence	NN	O	I-DNA
,	NN	O	O
an	NN	O	O
element	NN	O	O
located	NN	O	O
69	NN	O	B-DNA
bp	NN	O	I-DNA
upstream	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
IL4	NN	O	B-DNA
transcription	NN	O	I-DNA
initiation	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

Human	NN	O	B-DNA
IL4	NN	O	I-DNA
promoter	NN	O	I-DNA
-mediated	NN	O	O
transcription	NN	O	O
is	NN	O	O
downregulated	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
stimulated	NN	O	O
with	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B-activating	NN	O	I-protein
cytokine	NN	O	I-protein
tumor	NN	O	B-protein
necrosis	NN	O	I-protein
factor	NN	O	I-protein
alpha	NN	O	I-protein
and	NN	O	O
suppressed	NN	O	O
in	NN	O	O
RelA-overexpressing	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
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
stimulation	NN	O	O
or	NN	O	O
RelA	NN	O	B-protein
overexpression	NN	O	O
does	NN	O	O
not	NN	O	O
affect	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
a	NN	O	O
human	NN	O	B-DNA
IL4	NN	O	I-DNA
promoter	NN	O	I-DNA
containing	NN	O	O
a	NN	O	O
mouse	NN	O	B-DNA
P	NN	O	I-DNA
sequence	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
a	NN	O	O
higher-affinity	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
NF-ATp	NN	O	B-protein
and	NN	O	O
a	NN	O	O
lower-affinity	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
RelA	NN	O	B-protein
.	NN	O	O

Thus	NN	O	O
,	NN	O	O
competition	NN	O	O
between	NN	O	O
two	NN	O	O
general	NN	O	O
transcriptional	NN	O	B-protein
activators	NN	O	I-protein
,	NN	O	O
RelA	NN	O	B-protein
and	NN	O	O
NF-ATp	NN	O	B-protein
,	NN	O	O
mediates	NN	O	O
the	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
protein	NN	O	B-protein
kinase	NN	O	I-protein
C	NN	O	I-protein
stimulation	NN	O	O
on	NN	O	O
IL4	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
may	NN	O	O
contribute	NN	O	O
to	NN	O	O
differential	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
TH	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
the	NN	O	O
balance	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
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	O
(	NN	O	O
STAT	NN	O	B-protein
)	NN	O	O
transcription	NN	O	O
factor	NN	O	O
activity	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
inflammatory	NN	O	O
synovial	NN	O	O
fluids	NN	O	O
.	NN	O	O

The	NN	O	O
balance	NN	O	O
between	NN	O	O
type	NN	O	O
1	NN	O	O
and	NN	O	O
2	NN	O	O
T	NN	O	O
helper	NN	O	O
cell	NN	O	O
cytokine	NN	O	O
production	NN	O	O
plays	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
several	NN	O	O
animal	NN	O	O
models	NN	O	O
of	NN	O	O
autoimmunity	NN	O	O
,	NN	O	O
and	NN	O	O
skewed	NN	O	O
patterns	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
expression	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
human	NN	O	O
inflammatory	NN	O	O
diseases	NN	O	O
.	NN	O	O

Many	NN	O	O
cytokines	NN	O	B-protein
activate	NN	O	O
signal	NN	O	B-protein
transducer	NN	O	I-protein
and	NN	O	I-protein
activation	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
transcription	NN	O	I-protein
factors	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
activate	NN	O	O
transcription	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
effector	NN	O	O
genes	NN	O	O
.	NN	O	O

We	NN	O	O
used	NN	O	O
mononuclear	NN	O	B-cell_line
cell	NN	O	I-cell_line
priming	NN	O	I-cell_line
cultures	NN	O	I-cell_line
and	NN	O	O
inflammatory	NN	O	B-cell_type
synovial	NN	O	I-cell_type
fluids	NN	O	I-cell_type
(	NN	O	O
SFs	NN	O	B-cell_type
)	NN	O	O
derived	NN	O	O
from	NN	O	O
arthritis	NN	O	O
patients	NN	O	O
to	NN	O	O
examine	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
cytokine	NN	O	B-protein
production	NN	O	O
and	NN	O	O
STAT	NN	O	B-protein
activity	NN	O	O
by	NN	O	O
an	NN	O	O
inflammatory	NN	O	O
synovial	NN	O	O
microenvironment	NN	O	O
.	NN	O	O

Exposure	NN	O	O
to	NN	O	O
SFs	NN	O	B-cell_type
during	NN	O	O
priming	NN	O	O
resulted	NN	O	O
in	NN	O	O
an	NN	O	O
81	NN	O	O
%	NN	O	O
inhibition	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
-gamma	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	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
,	NN	O	O
production	NN	O	O
by	NN	O	O
effector	NN	O	B-cell_type
cells	NN	O	I-cell_type
generated	NN	O	O
in	NN	O	O
priming	NN	O	O
cultures	NN	O	O
.	NN	O	O

SF	NN	O	O
suppression	NN	O	O
was	NN	O	O
mediated	NN	O	O
by	NN	O	O
IL-4	NN	O	B-protein
and	NN	O	O
IL-10	NN	O	B-protein
and	NN	O	O
inhibition	NN	O	O
of	NN	O	O
IL-12	NN	O	B-protein
expression	NN	O	O
,	NN	O	O
and	NN	O	O
it	NN	O	O
was	NN	O	O
reversed	NN	O	O
in	NN	O	O
a	NN	O	O
dominant	NN	O	O
fashion	NN	O	O
by	NN	O	O
exogenous	NN	O	O
IL-12	NN	O	O
.	NN	O	O

SFs	NN	O	B-cell_type
blocked	NN	O	O
the	NN	O	O
sustained	NN	O	O
activity	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Stat1	NN	O	I-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
during	NN	O	O
the	NN	O	O
priming	NN	O	O
period	NN	O	O
,	NN	O	O
and	NN	O	O
Stat1	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
differentially	NN	O	O
regulated	NN	O	O
by	NN	O	O
cytokines	NN	O	B-protein
in	NN	O	O
parallel	NN	O	O
with	NN	O	O
their	NN	O	O
positive	NN	O	O
or	NN	O	O
negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
IFN-gamma	NN	O	B-protein
production	NN	O	O
.	NN	O	O

Active	NN	O	O
Stat3	NN	O	B-protein
,	NN	O	O
but	NN	O	O
not	NN	O	O
Stat1	NN	O	B-protein
,	NN	O	O
was	NN	O	O
detected	NN	O	O
in	NN	O	O
cells	NN	O	O
from	NN	O	O
inflamed	NN	O	O
joints	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
role	NN	O	O
for	NN	O	O
altered	NN	O	O
balance	NN	O	O
of	NN	O	O
Stat1	NN	O	O
and	NN	O	O
Stat3	NN	O	O
transcriptional	NN	O	O
activity	NN	O	O
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
differentiation	NN	O	O
and	NN	O	O
in	NN	O	O
the	NN	O	O
pathogenesis	NN	O	O
of	NN	O	O
inflammatory	NN	O	O
synovitis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Triggering	NN	O	O
of	NN	O	O
complement	NN	O	B-protein
receptors	NN	O	I-protein
CR1	NN	O	B-protein
(	NN	O	O
CD35	NN	O	B-protein
)	NN	O	O
and	NN	O	O
CR3	NN	O	B-protein
(	NN	O	O
CD11b/CD18	NN	O	B-protein
)	NN	O	O
induces	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
p50/p65	NN	O	B-protein
)	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
and	NN	O	O
enhances	NN	O	O
viral	NN	O	O
replication	NN	O	O
in	NN	O	O
HIV-infected	NN	O	B-cell_type
monocytic	NN	O	I-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Monocyte/macrophages	NN	O	B-cell_type
may	NN	O	O
harbor	NN	O	O
HIV	NN	O	O
in	NN	O	O
a	NN	O	O
nonproductive	NN	O	O
fashion	NN	O	O
for	NN	O	O
prolonged	NN	O	O
periods	NN	O	O
of	NN	O	O
time	NN	O	O
.	NN	O	O

Viral	NN	O	O
gene	NN	O	O
expression	NN	O	O
may	NN	O	O
be	NN	O	O
reactivated	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
LPS	NN	O	O
or	NN	O	O
cytokines	NN	O	B-protein
such	NN	O	O
as	NN	O	O
TNF-alpha	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

The	NN	O	O
effect	NN	O	O
of	NN	O	O
LPS	NN	O	O
and	NN	O	O
TNF-alpha	NN	O	B-protein
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
nuclear	NN	O	O
translocation	NN	O	O
of	NN	O	O
the	NN	O	O
DNA-binding	NN	O	B-protein
heterodimer	NN	O	I-protein
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
(	NN	O	O
p50/p65	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
a	NN	O	O
specific	NN	O	O
sequence	NN	O	O
in	NN	O	O
the	NN	O	O
HIV-long	NN	O	B-DNA
terminal	NN	O	I-DNA
repeat	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
present	NN	O	O
study	NN	O	O
demonstrates	NN	O	O
that	NN	O	O
triggering	NN	O	O
of	NN	O	O
complement	NN	O	B-protein
receptors	NN	O	I-protein
CR1	NN	O	B-protein
(	NN	O	O
CD35	NN	O	B-protein
)	NN	O	O
and	NN	O	O
CR3	NN	O	B-protein
(	NN	O	O
CD11b/CD18	NN	O	B-protein
)	NN	O	O
enhances	NN	O	O
viral	NN	O	O
replication	NN	O	O
in	NN	O	O
HIV-infected	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

Monocytic	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
normal	NN	O	O
peripheral	NN	O	O
blood	NN	O	O
monocytes	NN	O	O
were	NN	O	O
infected	NN	O	O
with	NN	O	O
HIV-1	NN	O	O
in	NN	O	O
vitro	NN	O	O
and	NN	O	O
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
F	NN	O	B-protein
(	NN	O	I-protein
ab	NN	O	I-protein
'	NN	O	I-protein
)	NN	O	I-protein
2	NN	O	I-protein
fragments	NN	O	I-protein
of	NN	O	O
monoclonal	NN	O	B-protein
anti-CR1	NN	O	I-protein
or	NN	O	O
anti-CR3	NN	O	B-protein
Abs	NN	O	I-protein
or	NN	O	O
with	NN	O	O
C3	NN	O	B-protein
fragments	NN	O	I-protein
.	NN	O	O

Stimulation	NN	O	O
of	NN	O	O
CR1	NN	O	B-protein
or	NN	O	O
CR3	NN	O	B-protein
induces	NN	O	O
a	NN	O	O
two-	NN	O	O
to	NN	O	O
fourfold	NN	O	O
increase	NN	O	O
in	NN	O	O
the	NN	O	O
amount	NN	O	O
of	NN	O	O
cell-associated	NN	O	O
and	NN	O	O
released	NN	O	O
p24	NN	O	B-protein
Ag	NN	O	I-protein
in	NN	O	O
cell	NN	O	B-cell_line
cultures	NN	O	I-cell_line
that	NN	O	O
was	NN	O	O
equivalent	NN	O	O
to	NN	O	O
that	NN	O	O
observed	NN	O	O
in	NN	O	O
control	NN	O	B-cell_line
cultures	NN	O	I-cell_line
triggered	NN	O	O
with	NN	O	O
LPS	NN	O	O
.	NN	O	O

We	NN	O	O
further	NN	O	O
observed	NN	O	O
that	NN	O	O
stimulation	NN	O	O
of	NN	O	O
CR1	NN	O	B-protein
or	NN	O	O
CR3	NN	O	B-protein
induces	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	NN	O	I-protein
p50/p65	NN	O	B-protein
in	NN	O	O
infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Translocation	NN	O	O
of	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
p50/p65	NN	O	B-protein
was	NN	O	O
also	NN	O	O
observed	NN	O	O
following	NN	O	O
stimulation	NN	O	O
of	NN	O	O
CR1	NN	O	B-protein
or	NN	O	O
CR3	NN	O	B-protein
of	NN	O	O
uninfected	NN	O	B-cell_type
peripheral	NN	O	I-cell_type
blood	NN	O	I-cell_type
monocytes	NN	O	I-cell_type
from	NN	O	O
HIV-seronegative	NN	O	O
donors	NN	O	O
.	NN	O	O

The	NN	O	O
amount	NN	O	O
of	NN	O	O
protein	NN	O	O
translocated	NN	O	O
was	NN	O	O
similar	NN	O	O
to	NN	O	O
that	NN	O	O
observed	NN	O	O
when	NN	O	O
cells	NN	O	O
were	NN	O	O
stimulated	NN	O	O
with	NN	O	O
rhTNF-alpha	NN	O	B-protein
.	NN	O	O

TNF-alpha	NN	O	B-protein
did	NN	O	O
not	NN	O	O
mediate	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
p50/p65	NN	O	B-protein
induced	NN	O	O
by	NN	O	O
triggering	NN	O	O
of	NN	O	O
complement	NN	O	B-protein
receptors	NN	O	I-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
HIV	NN	O	O
gene	NN	O	O
expression	NN	O	O
may	NN	O	O
be	NN	O	O
activated	NN	O	O
in	NN	O	O
infected	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
through	NN	O	O
interaction	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
complement-opsonized	NN	O	B-protein
particles	NN	O	I-protein
and	NN	O	O
that	NN	O	O
enhanced	NN	O	O
viral	NN	O	O
replication	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
C3	NN	O	B-protein
receptor	NN	O	I-protein
-mediated	NN	O	O
nuclear	NN	O	O
translocation	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

Attenuation	NN	O	O
of	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
-induced	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
infected	NN	O	O
with	NN	O	O
Leishmania	NN	O	O
donovani	NN	O	O
:	NN	O	O
selective	NN	O	O
inhibition	NN	O	O
of	NN	O	O
signaling	NN	O	O
through	NN	O	O
Janus	NN	O	B-protein
kinases	NN	O	I-protein
and	NN	O	O
Stat1	NN	O	B-protein
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
gene	NN	O	O
transcription	NN	O	O
in	NN	O	O
response	NN	O	O
to	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
is	NN	O	O
impaired	NN	O	O
in	NN	O	O
mononuclear	NN	O	B-cell_type
phagocytes	NN	O	I-cell_type
infected	NN	O	O
with	NN	O	O
Leishmania	NN	O	O
donovani	NN	O	O
,	NN	O	O
and	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
involved	NN	O	O
are	NN	O	O
not	NN	O	O
fully	NN	O	O
understood	NN	O	O
.	NN	O	O

The	NN	O	O
changes	NN	O	O
in	NN	O	O
gene	NN	O	O
expression	NN	O	O
brought	NN	O	O
about	NN	O	O
by	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
are	NN	O	O
thought	NN	O	O
to	NN	O	O
involve	NN	O	O
transient	NN	O	O
increases	NN	O	O
in	NN	O	O
the	NN	O	O
activities	NN	O	O
of	NN	O	O
cellular	NN	O	B-protein
protein	NN	O	I-protein
tyrosine	NN	O	I-protein
kinases	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
Jak2	NN	O	B-protein
,	NN	O	O
leading	NN	O	O
to	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
Stat1	NN	O	I-protein
.	NN	O	O

To	NN	O	O
investigate	NN	O	O
the	NN	O	O
mechanisms	NN	O	O
accounting	NN	O	O
for	NN	O	O
the	NN	O	O
impaired	NN	O	O
responses	NN	O	O
to	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
,	NN	O	O
a	NN	O	O
model	NN	O	O
system	NN	O	O
for	NN	O	O
examining	NN	O	O
overall	NN	O	O
changes	NN	O	O
in	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
,	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
and	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Stat1	NN	O	B-protein
was	NN	O	O
developed	NN	O	O
in	NN	O	O
phorbol	NN	O	B-cell_line
12-myristate	NN	O	I-cell_line
13-acetate-differentiated	NN	O	I-cell_line
U-937	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Analysis	NN	O	O
of	NN	O	O
whole-cell	NN	O	O
lysates	NN	O	O
by	NN	O	O
antiphosphotyrosine	NN	O	O
immunoblotting	NN	O	O
showed	NN	O	O
that	NN	O	O
incubation	NN	O	O
with	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
brought	NN	O	O
about	NN	O	O
specific	NN	O	O
increases	NN	O	O
in	NN	O	O
phosphotyrosine	NN	O	O
labeling	NN	O	O
of	NN	O	O
several	NN	O	O
proteins	NN	O	O
.	NN	O	O

Increased	NN	O	O
labeling	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
occurred	NN	O	O
to	NN	O	O
similar	NN	O	O
extents	NN	O	O
in	NN	O	O
control	NN	O	B-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
cells	NN	O	O
that	NN	O	O
had	NN	O	O
been	NN	O	O
infected	NN	O	O
with	NN	O	O
L.	NN	O	O
donovani	NN	O	O
for	NN	O	O
16	NN	O	O
h	NN	O	O
.	NN	O	O

Jak1	NN	O	B-protein
,	NN	O	O
Jak2	NN	O	B-protein
,	NN	O	O
and	NN	O	O
Stat1	NN	O	B-protein
were	NN	O	O
immunoprecipitated	NN	O	O
from	NN	O	O
control	NN	O	O
and	NN	O	O
interferon-treated	NN	O	B-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
these	NN	O	O
proteins	NN	O	O
,	NN	O	O
detected	NN	O	O
by	NN	O	O
antiphosphotyrosine	NN	O	O
immunoblotting	NN	O	O
was	NN	O	O
used	NN	O	O
to	NN	O	O
measured	NN	O	O
their	NN	O	O
activation	NN	O	O
.	NN	O	O

Tyrosine	NN	O	O
phosphorylation	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
Stat1	NN	O	B-protein
increased	NN	O	O
markedly	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
in	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
incubated	NN	O	O
with	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
.	NN	O	O

In	NN	O	O
contrast	NN	O	O
,	NN	O	O
in	NN	O	O
cells	NN	O	O
infected	NN	O	O
with	NN	O	O
L.	NN	O	O
donovani	NN	O	O
,	NN	O	O
tyrosine	NN	O	O
phosphorylation	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
Stat1	NN	O	B-protein
was	NN	O	O
markedly	NN	O	O
impaired	NN	O	O
.	NN	O	O

This	NN	O	O
effect	NN	O	O
was	NN	O	O
dependent	NN	O	O
upon	NN	O	O
the	NN	O	O
duration	NN	O	O
of	NN	O	O
exposure	NN	O	O
to	NN	O	O
L.	NN	O	O
donovani	NN	O	O
and	NN	O	O
was	NN	O	O
maximal	NN	O	O
and	NN	O	O
complete	NN	O	O
at	NN	O	O
16	NN	O	O
h	NN	O	O
.	NN	O	O

Results	NN	O	O
similar	NN	O	O
to	NN	O	O
those	NN	O	O
observed	NN	O	O
with	NN	O	O
U-937	NN	O	B-cell_line
cells	NN	O	I-cell_line
were	NN	O	O
also	NN	O	O
obtained	NN	O	O
with	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

These	NN	O	O
findings	NN	O	O
indicate	NN	O	O
that	NN	O	O
infection	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
mononuclear	NN	O	I-cell_type
phagocytes	NN	O	I-cell_type
with	NN	O	O
L.	NN	O	O
donovani	NN	O	O
leads	NN	O	O
to	NN	O	O
impaired	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
-mediated	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
selective	NN	O	O
effects	NN	O	O
on	NN	O	O
the	NN	O	O
Jak-	NN	O	O
Stat1	NN	O	B-protein
pathway	NN	O	O
.	NN	O	O

Unresponsiveness	NN	O	O
to	NN	O	O
gamma	NN	O	B-protein
interferon	NN	O	I-protein
for	NN	O	O
activation	NN	O	O
of	NN	O	O
this	NN	O	O
pathway	NN	O	O
may	NN	O	O
explain	NN	O	O
impaired	NN	O	O
transcriptional	NN	O	O
responses	NN	O	O
in	NN	O	O
leishmania-infected	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Mutually	NN	O	O
exclusive	NN	O	O
interaction	NN	O	O
of	NN	O	O
a	NN	O	O
novel	NN	O	O
matrix	NN	O	B-protein
attachment	NN	O	I-protein
region	NN	O	I-protein
binding	NN	O	I-protein
protein	NN	O	I-protein
and	NN	O	O
the	NN	O	O
NF-muNR	NN	O	B-protein
enhancer	NN	O	I-protein
repressor	NN	O	I-protein
.	NN	O	O

Implications	NN	O	O
for	NN	O	O
regulation	NN	O	O
of	NN	O	O
immunoglobulin	NN	O	B-protein
heavy	NN	O	I-protein
chain	NN	O	I-protein
expression	NN	O	O
.	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
intronic	NN	O	O
enhancer	NN	O	O
stimulates	NN	O	O
transcription	NN	O	O
from	NN	O	O
functional	NN	O	O
promoters	NN	O	O
in	NN	O	O
B	NN	O	O
lymphocytes	NN	O	O
but	NN	O	O
not	NN	O	O
other	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

The	NN	O	O
observation	NN	O	O
that	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
factor-mu	NN	O	I-protein
negative	NN	O	I-protein
regulator	NN	O	I-protein
(	NN	O	I-protein
NF-muNR	NN	O	I-protein
)	NN	O	I-protein
enhancer	NN	O	I-protein
repressor	NN	O	I-protein
overlap	NN	O	I-protein
nuclear	NN	O	I-protein
matrix	NN	O	I-protein
attachment	NN	O	I-protein
regions	NN	O	I-protein
(	NN	O	O
MARs	NN	O	B-protein
)	NN	O	O
in	NN	O	O
this	NN	O	O
enhancer	NN	O	B-DNA
has	NN	O	O
lead	NN	O	O
to	NN	O	O
the	NN	O	O
hypothesis	NN	O	O
that	NN	O	O
the	NN	O	O
cell	NN	O	O
type	NN	O	O
specificity	NN	O	O
of	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
might	NN	O	O
be	NN	O	O
controlled	NN	O	O
by	NN	O	O
regulating	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
attachment	NN	O	O
(	NN	O	O
Scheuermann	NN	O	O
,	NN	O	O
R.	NN	O	O
H.	NN	O	O
,	NN	O	O
and	NN	O	O
Chen	NN	O	O
,	NN	O	O
U.	NN	O	O
(	NN	O	O
1989	NN	O	O
)	NN	O	O
Genes	NN	O	O
&	NN	O	O
Dev.	NN	O	O
3	NN	O	O
,	NN	O	O
1255-1266	NN	O	O
)	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
MARs	NN	O	B-protein
in	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
regulation	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	O
MAR-binding	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
MAR-BP1	NN	O	B-protein
,	NN	O	O
from	NN	O	O
soluble	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
preparations	NN	O	O
based	NN	O	O
on	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
bind	NN	O	O
to	NN	O	O
the	NN	O	O
MARs	NN	O	B-protein
associated	NN	O	O
with	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
.	NN	O	O

Purified	NN	O	O
MAR-BP1	NN	O	B-protein
migrates	NN	O	O
as	NN	O	O
a	NN	O	O
33-kDa	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
and	NN	O	O
it	NN	O	O
can	NN	O	O
be	NN	O	O
found	NN	O	O
in	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
preparations	NN	O	O
from	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
different	NN	O	O
types	NN	O	O
of	NN	O	O
lymphoid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Although	NN	O	O
specific	NN	O	O
binding	NN	O	O
sites	NN	O	O
have	NN	O	O
been	NN	O	O
difficult	NN	O	O
to	NN	O	O
localize	NN	O	O
by	NN	O	O
chemical	NN	O	O
or	NN	O	O
enzymatic	NN	O	O
footprinting	NN	O	O
procedures	NN	O	O
,	NN	O	O
NF-muNR	NN	O	B-protein
binding	NN	O	O
sites	NN	O	O
are	NN	O	O
critical	NN	O	O
for	NN	O	O
efficient	NN	O	O
MAR-BP1	NN	O	B-protein
binding	NN	O	O
.	NN	O	O

Indeed	NN	O	O
,	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
to	NN	O	O
either	NN	O	O
intact	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
preparations	NN	O	O
or	NN	O	O
to	NN	O	O
MAR-BP1	NN	O	B-protein
is	NN	O	O
mutually	NN	O	O
exclusive	NN	O	O
to	NN	O	O
NF-muNR	NN	O	B-protein
binding	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
model	NN	O	O
for	NN	O	O
cell-type	NN	O	O
specific	NN	O	O
regulation	NN	O	O
in	NN	O	O
which	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
NF-muNR	NN	O	B-protein
repressor	NN	O	I-protein
to	NN	O	O
the	NN	O	O
IgH	NN	O	B-DNA
enhancer	NN	O	I-DNA
prevents	NN	O	O
nuclear	NN	O	O
matrix	NN	O	O
attachment	NN	O	O
in	NN	O	O
inappropriate	NN	O	O
cells	NN	O	O
by	NN	O	O
interfering	NN	O	O
with	NN	O	O
MAR-BP1	NN	O	B-protein
/enhancer	NN	O	B-DNA
interaction	NN	O	O
.	NN	O	O

-DOCSTART-	O

PU.1	NN	O	B-protein
(	NN	O	O
Spi-1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
regulate	NN	O	O
expression	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
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	NN	O	O

Growth	NN	O	B-protein
factor	NN	O	I-protein
receptors	NN	O	I-protein
play	NN	O	O
an	NN	O	O
important	NN	O	O
role	NN	O	O
in	NN	O	O
hematopoiesis	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
mechanisms	NN	O	O
directing	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
key	NN	O	O
regulators	NN	O	O
of	NN	O	O
hematopoiesis	NN	O	O
,	NN	O	O
we	NN	O	O
initiated	NN	O	O
a	NN	O	O
study	NN	O	O
investigating	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
activating	NN	O	O
the	NN	O	O
expression	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
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
gene	NN	O	I-DNA
.	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
human	NN	O	B-DNA
GM-CSF	NN	O	I-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
directs	NN	O	O
reporter	NN	O	B-DNA
gene	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
a	NN	O	O
tissue-specific	NN	O	O
fashion	NN	O	O
in	NN	O	O
myelomonocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
which	NN	O	O
correlates	NN	O	O
with	NN	O	O
its	NN	O	O
expression	NN	O	O
pattern	NN	O	O
as	NN	O	O
analyzed	NN	O	O
by	NN	O	O
reverse	NN	O	O
transcription	NN	O	O
PCR	NN	O	O
.	NN	O	O

The	NN	O	O
GM-CSF	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
contains	NN	O	O
an	NN	O	O
important	NN	O	O
functional	NN	O	O
site	NN	O	O
between	NN	O	O
positions	NN	O	O
-53	NN	O	O
and	NN	O	O
-41	NN	O	O
as	NN	O	O
identified	NN	O	O
by	NN	O	O
deletion	NN	O	O
analysis	NN	O	O
of	NN	O	O
reporter	NN	O	B-DNA
constructs	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
myeloid	NN	O	O
and	NN	O	O
B	NN	O	B-protein
cell	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
PU.1	NN	O	B-protein
binds	NN	O	O
specifically	NN	O	O
to	NN	O	O
this	NN	O	O
site	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
a	NN	O	O
CCAAT	NN	O	B-DNA
site	NN	O	I-DNA
located	NN	O	O
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
PU.1	NN	O	B-DNA
site	NN	O	I-DNA
between	NN	O	O
positions	NN	O	O
-70	NN	O	O
and	NN	O	O
-54	NN	O	O
is	NN	O	O
involved	NN	O	O
in	NN	O	O
positive-negative	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
is	NN	O	O
the	NN	O	O
major	NN	O	O
CCAAT/enhancer-binding	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
C/EBP	NN	O	B-protein
)	NN	O	O
form	NN	O	O
binding	NN	O	O
to	NN	O	O
this	NN	O	O
site	NN	O	O
in	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
of	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Point	NN	O	O
mutations	NN	O	O
of	NN	O	O
either	NN	O	O
the	NN	O	O
PU.1	NN	O	B-DNA
site	NN	O	I-DNA
or	NN	O	O
the	NN	O	O
C/EBP	NN	O	B-DNA
site	NN	O	I-DNA
that	NN	O	O
abolish	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
respective	NN	O	O
factors	NN	O	O
result	NN	O	O
in	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
of	NN	O	O
GM-CSF	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
in	NN	O	O
myelomonocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
only	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
in	NN	O	O
myeloid	NN	O	O
and	NN	O	O
B	NN	O	O
cell	NN	O	O
extracts	NN	O	O
,	NN	O	O
PU.1	NN	O	B-protein
forms	NN	O	O
a	NN	O	O
novel	NN	O	O
,	NN	O	O
specific	NN	O	O
,	NN	O	O
more	NN	O	O
slowly	NN	O	O
migrating	NN	O	O
complex	NN	O	O
(	NN	O	O
PU-SF	NN	O	B-protein
)	NN	O	O
when	NN	O	O
binding	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-protein
receptor	NN	O	I-protein
alpha	NN	O	I-protein
promoter	NN	O	O
PU.1	NN	O	B-DNA
site	NN	O	I-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
specific	NN	O	O
interaction	NN	O	O
with	NN	O	O
PU.1	NN	O	B-protein
on	NN	O	O
a	NN	O	O
myeloid	NN	O	B-DNA
PU.1	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
novel	NN	O	O
complex	NN	O	O
is	NN	O	O
distinct	NN	O	O
from	NN	O	O
that	NN	O	O
described	NN	O	O
previously	NN	O	O
as	NN	O	O
binding	NN	O	O
to	NN	O	O
B	NN	O	B-DNA
cell	NN	O	I-DNA
enhancer	NN	O	I-DNA
sites	NN	O	I-DNA
and	NN	O	O
can	NN	O	O
be	NN	O	O
formed	NN	O	O
by	NN	O	O
addition	NN	O	O
of	NN	O	O
PU.1	NN	O	B-protein
to	NN	O	O
extracts	NN	O	O
from	NN	O	O
certain	NN	O	O
nonmyeloid	NN	O	B-cell_type
cell	NN	O	I-cell_type
types	NN	O	I-cell_type
which	NN	O	O
do	NN	O	O
not	NN	O	O
express	NN	O	O
PU.1	NN	O	B-protein
,	NN	O	O
including	NN	O	O
T	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
but	NN	O	O
not	NN	O	O
from	NN	O	O
erythroid	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
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
PU-SF	NN	O	B-protein
complex	NN	O	I-protein
binds	NN	O	O
to	NN	O	O
PU.1	NN	O	B-protein
sites	NN	O	O
found	NN	O	O
on	NN	O	O
a	NN	O	O
number	NN	O	O
of	NN	O	O
myeloid	NN	O	B-DNA
promoters	NN	O	I-DNA
,	NN	O	O
and	NN	O	O
its	NN	O	O
formation	NN	O	O
requires	NN	O	O
an	NN	O	O
intact	NN	O	O
PU.1	NN	O	B-DNA
site	NN	O	I-DNA
adjacent	NN	O	O
to	NN	O	O
a	NN	O	O
single-stranded	NN	O	O
region	NN	O	O
.	NN	O	O

Expression	NN	O	O
of	NN	O	O
PU.1	NN	O	B-protein
in	NN	O	O
nonmyeloid	NN	O	B-cell_type
cells	NN	O	I-cell_type
can	NN	O	O
activate	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Deletion	NN	O	O
of	NN	O	O
the	NN	O	O
amino-terminal	NN	O	B-protein
region	NN	O	I-protein
of	NN	O	O
PU.1	NN	O	B-protein
results	NN	O	O
in	NN	O	O
a	NN	O	O
failure	NN	O	O
to	NN	O	O
form	NN	O	O
the	NN	O	O
PU-SF	NN	O	B-protein
complex	NN	O	O
and	NN	O	O
in	NN	O	O
a	NN	O	O
concomitant	NN	O	O
loss	NN	O	O
of	NN	O	O
transactivation	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
formation	NN	O	O
of	NN	O	O
the	NN	O	O
PU-SF	NN	O	B-protein
complex	NN	O	I-protein
is	NN	O	O
of	NN	O	O
functional	NN	O	O
importance	NN	O	O
for	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

Finally	NN	O	O
,	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
C/EBP	NN	O	B-protein
alpha	NN	O	I-protein
can	NN	O	O
also	NN	O	O
active	NN	O	O
the	NN	O	O
GM-CSF	NN	O	B-DNA
receptor	NN	O	I-DNA
alpha	NN	O	I-DNA
promoter	NN	O	I-DNA
in	NN	O	O
nonmyeloid	NN	O	B-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
400	NN	O	O
WORDS	NN	O	O
)	NN	O	O

-DOCSTART-	O

HMG-I	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
GATA	NN	O	B-DNA
motifs	NN	O	I-DNA
:	NN	O	O
implications	NN	O	O
for	NN	O	O
an	NN	O	O
HPFH	NN	O	O
syndrome	NN	O	O
.	NN	O	O

We	NN	O	O
have	NN	O	O
examined	NN	O	O
binding	NN	O	O
of	NN	O	O
the	NN	O	O
nuclear	NN	O	B-protein
protein	NN	O	I-protein
HMG-I	NN	O	B-protein
to	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
gamma-globin	NN	O	I-DNA
promoter	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
find	NN	O	O
that	NN	O	O
HMG-I	NN	O	B-protein
binds	NN	O	O
preferentially	NN	O	O
to	NN	O	O
the	NN	O	O
more	NN	O	O
3	NN	O	O
'	NN	O	O
of	NN	O	O
a	NN	O	O
pair	NN	O	O
of	NN	O	O
GATA	NN	O	B-DNA
motifs	NN	O	I-DNA
in	NN	O	O
the	NN	O	O
gamma-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
;	NN	O	O
this	NN	O	O
paired	NN	O	O
motif	NN	O	O
is	NN	O	O
bound	NN	O	O
by	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

A	NN	O	O
naturally	NN	O	O
occurring	NN	O	O
mutation	NN	O	O
(	NN	O	O
-175	NN	O	O
T-C	NN	O	O
)	NN	O	O
in	NN	O	O
the	NN	O	O
area	NN	O	O
bound	NN	O	O
by	NN	O	O
HMG-I	NN	O	B-protein
results	NN	O	O
in	NN	O	O
overexpression	NN	O	O
of	NN	O	O
gamma-globin	NN	O	B-protein
in	NN	O	O
adult	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
HPFH	NN	O	B-cell_type
)	NN	O	O
and	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
the	NN	O	O
gamma-globin	NN	O	B-DNA
promoter	NN	O	I-DNA
in	NN	O	O
in	NN	O	O
vitro	NN	O	O
expression	NN	O	O
assays	NN	O	O
;	NN	O	O
HMG-I	NN	O	B-protein
does	NN	O	O
not	NN	O	O
bind	NN	O	O
to	NN	O	O
this	NN	O	O
mutant	NN	O	B-DNA
sequence	NN	O	I-DNA
.	NN	O	O

A	NN	O	O
survey	NN	O	O
of	NN	O	O
GATA	NN	O	B-DNA
motifs	NN	O	I-DNA
from	NN	O	O
other	NN	O	O
globin	NN	O	B-DNA
cis-elements	NN	O	I-DNA
demonstrates	NN	O	O
HMG-I	NN	O	B-protein
binding	NN	O	O
to	NN	O	O
most	NN	O	O
of	NN	O	O
them	NN	O	O
.	NN	O	O

These	NN	O	O
findings	NN	O	O
implicate	NN	O	O
HMG-I	NN	O	B-protein
in	NN	O	O
the	NN	O	O
HPFH	NN	O	B-cell_type
phenotype	NN	O	O
;	NN	O	O
we	NN	O	O
speculate	NN	O	O
that	NN	O	O
it	NN	O	O
may	NN	O	O
participate	NN	O	O
in	NN	O	O
the	NN	O	O
formation	NN	O	O
of	NN	O	O
multiprotein	NN	O	B-protein
complexes	NN	O	I-protein
that	NN	O	O
regulate	NN	O	O
globin	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Constitutive	NN	O	O
activation	NN	O	O
of	NN	O	O
different	NN	O	O
Jak	NN	O	B-protein
tyrosine	NN	O	I-protein
kinases	NN	O	I-protein
in	NN	O	O
human	NN	O	B-protein
T	NN	O	I-protein
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
protein	NN	O	I-protein
or	NN	O	O
virus-transformed	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

HTLV-1	NN	O	O
infection	NN	O	O
causes	NN	O	O
an	NN	O	O
adult	NN	O	O
T	NN	O	O
cell	NN	O	O
leukemia	NN	O	O
in	NN	O	O
humans	NN	O	O
.	NN	O	O

The	NN	O	O
viral	NN	O	B-protein
encoded	NN	O	I-protein
protein	NN	O	I-protein
tax	NN	O	B-protein
,	NN	O	O
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
oncogenesis	NN	O	O
.	NN	O	O

Our	NN	O	O
previous	NN	O	O
data	NN	O	O
obtained	NN	O	O
from	NN	O	O
a	NN	O	O
tax	NN	O	B-protein
transgenic	NN	O	O
mouse	NN	O	O
model	NN	O	O
revealed	NN	O	O
that	NN	O	O
tax	NN	O	B-protein
transforms	NN	O	O
mouse	NN	O	B-cell_type
fibroblasts	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
thymocytes	NN	O	B-cell_type
,	NN	O	O
despite	NN	O	O
comparable	NN	O	O
levels	NN	O	O
of	NN	O	O
tax	NN	O	B-protein
expression	NN	O	O
in	NN	O	O
both	NN	O	O
tissues	NN	O	O
.	NN	O	O

Constitutive	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
a	NN	O	O
130-kD	NN	O	B-protein
protein	NN	O	I-protein
(	NN	O	O
s	NN	O	O
)	NN	O	O
was	NN	O	O
observed	NN	O	O
in	NN	O	O
the	NN	O	O
tax	NN	O	B-protein
transformed	NN	O	B-cell_line
fibroblast	NN	O	I-cell_line
B	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
in	NN	O	O
HTLV-1	NN	O	B-cell_line
transformed	NN	O	I-cell_line
human	NN	O	I-cell_line
lymphoid	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
thymocytes	NN	O	B-cell_type
from	NN	O	O
Thy-tax	NN	O	O
transgenic	NN	O	O
mice	NN	O	O
.	NN	O	O

Phosphotyrosine	NN	O	O
immunoprecipitation	NN	O	O
followed	NN	O	O
by	NN	O	O
Western	NN	O	O
blot	NN	O	O
analysis	NN	O	O
with	NN	O	O
a	NN	O	O
set	NN	O	O
of	NN	O	O
Jak	NN	O	B-protein
kinase	NN	O	I-protein
specific	NN	O	I-protein
antibodies	NN	O	I-protein
,	NN	O	O
identified	NN	O	O
p130	NN	O	O
as	NN	O	O
Jak2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
tax	NN	O	B-protein
transformed	NN	O	O
mouse	NN	O	B-cell_line
fibroblastic	NN	O	I-cell_line
cell	NN	O	I-cell_line
line	NN	O	I-cell_line
and	NN	O	O
Jak3	NN	O	B-protein
in	NN	O	O
HTLV-1	NN	O	B-cell_line
transformed	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

Phosphorylation	NN	O	O
of	NN	O	O
Jak2	NN	O	B-protein
in	NN	O	O
tax	NN	O	B-cell_line
transformed	NN	O	I-cell_line
cells	NN	O	I-cell_line
resulted	NN	O	O
from	NN	O	O
high	NN	O	O
expression	NN	O	O
of	NN	O	O
IL-6	NN	O	O
.	NN	O	O

Tyrosine	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
this	NN	O	O
protein	NN	O	O
could	NN	O	O
also	NN	O	O
be	NN	O	O
induced	NN	O	O
in	NN	O	O
Balb/c3T3	NN	O	B-cell_line
cells	NN	O	I-cell_line
using	NN	O	O
a	NN	O	O
supernatant	NN	O	O
from	NN	O	O
the	NN	O	O
B	NN	O	B-cell_line
line	NN	O	I-cell_line
,	NN	O	O
which	NN	O	O
was	NN	O	O
associated	NN	O	O
with	NN	O	O
induction	NN	O	O
of	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
.	NN	O	O

Both	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
proliferation	NN	O	O
were	NN	O	O
inhibited	NN	O	O
by	NN	O	O
IL-6	NN	O	B-protein
neutralizing	NN	O	I-protein
antibodies	NN	O	I-protein
.	NN	O	O

Constitutive	NN	O	O
phosphorylation	NN	O	O
of	NN	O	O
Jak	NN	O	B-protein
kinases	NN	O	I-protein
may	NN	O	O
facilitate	NN	O	O
tumor	NN	O	O
growth	NN	O	O
in	NN	O	O
both	NN	O	O
HTLV-1	NN	O	B-cell_type
infected	NN	O	I-cell_type
human	NN	O	I-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
and	NN	O	O
the	NN	O	O
transgenic	NN	O	O
mouse	NN	O	O
model	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
by	NN	O	O
hydroxyurea	NN	O	O
in	NN	O	O
human	NN	O	B-cell_line
K562	NN	O	I-cell_line
cells	NN	O	I-cell_line
during	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
[	NN	O	O
published	NN	O	O
erratum	NN	O	O
appears	NN	O	O
in	NN	O	O
Biochim	NN	O	O
Biophys	NN	O	O
Acta	NN	O	O
1995	NN	O	O
Dec	NN	O	O
27	NN	O	O
;	NN	O	O
1264	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
:	NN	O	O
409	NN	O	O
]	NN	O	O

Hydroxyurea	NN	O	O
(	NN	O	O
HU	NN	O	O
)	NN	O	O
is	NN	O	O
an	NN	O	O
antitumor	NN	O	O
agent	NN	O	O
which	NN	O	O
also	NN	O	O
induces	NN	O	O
hemoglobinization	NN	O	O
during	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
HU	NN	O	O
stimulates	NN	O	O
the	NN	O	O
synthesis	NN	O	O
of	NN	O	O
fetal	NN	O	B-protein
hemoglobin	NN	O	I-protein
in	NN	O	O
sickle	NN	O	O
cell	NN	O	O
anemia	NN	O	O
patients	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
understand	NN	O	O
its	NN	O	O
mechanism	NN	O	O
of	NN	O	O
action	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
HU	NN	O	O
on	NN	O	O
regulation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
expression	NN	O	O
prior	NN	O	O
to	NN	O	O
the	NN	O	O
onset	NN	O	O
of	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

HU	NN	O	O
induced	NN	O	O
a	NN	O	O
dose-dependent	NN	O	O
stimulation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
synthesis	NN	O	O
.	NN	O	O

The	NN	O	O
levels	NN	O	O
of	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
elevated	NN	O	O
4	NN	O	O
to	NN	O	O
7.5-fold	NN	O	O
by	NN	O	O
HU	NN	O	O
within	NN	O	O
2	NN	O	O
h	NN	O	O
.	NN	O	O

This	NN	O	O
was	NN	O	O
followed	NN	O	O
by	NN	O	O
a	NN	O	O
gradual	NN	O	O
decline	NN	O	O
to	NN	O	O
the	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

Both	NN	O	O
nuclear	NN	O	O
run-on	NN	O	O
and	NN	O	O
actinomycin	NN	O	O
D	NN	O	O
pulse	NN	O	O
experiments	NN	O	O
strongly	NN	O	O
indicate	NN	O	O
that	NN	O	O
HU	NN	O	O
regulates	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
expression	NN	O	O
by	NN	O	O
increasing	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
synthesis	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
stabilizing	NN	O	O
the	NN	O	O
c-jun	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	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
jun	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
elevated	NN	O	O
by	NN	O	O
2	NN	O	O
to	NN	O	O
5-fold	NN	O	O
within	NN	O	O
4	NN	O	O
h	NN	O	O
in	NN	O	O
HU	NN	O	B-cell_line
treated	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
concentrations	NN	O	O
of	NN	O	O
HU	NN	O	O
below	NN	O	O
250	NN	O	O
microM	NN	O	O
slightly	NN	O	O
increased	NN	O	O
the	NN	O	O
5X	NN	O	O
AP-1	NN	O	B-protein
/CAT	NN	O	B-protein
activity	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
strongly	NN	O	O
suggest	NN	O	O
that	NN	O	O
HU	NN	O	O
induces	NN	O	O
both	NN	O	O
transcriptional	NN	O	O
and	NN	O	O
post-transcription	NN	O	O
regulation	NN	O	O
of	NN	O	O
c-jun	NN	O	B-DNA
during	NN	O	O
erythroid	NN	O	O
differentiation	NN	O	O
.	NN	O	O

-DOCSTART-	O

In	NN	O	O
vivo	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
effects	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
on	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
in	NN	O	O
man	NN	O	O
:	NN	O	O
relationship	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	B-protein
receptors	NN	O	I-protein
.	NN	O	O

Interrelations	NN	O	O
between	NN	O	O
the	NN	O	O
hypothalamic-pituitary-adrenal	NN	O	O
system	NN	O	O
(	NN	O	O
HPA	NN	O	O
)	NN	O	O
and	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
represent	NN	O	O
a	NN	O	O
well-documented	NN	O	O
biological	NN	O	O
phenomenon	NN	O	O
.	NN	O	O

While	NN	O	O
in	NN	O	O
vitro	NN	O	O
administration	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
may	NN	O	O
inhibit	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
-	NN	O	O
and	NN	O	O
phytohemagglutinin	NN	O	O
(	NN	O	O
PHA	NN	O	O
)	NN	O	O
-induced	NN	O	O
T-cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
pokeweed	NN	O	B-protein
mitogen	NN	O	I-protein
(	NN	O	O
PWM	NN	O	B-protein
)	NN	O	O
-driven	NN	O	O
B-cell	NN	O	O
mitogenesis	NN	O	O
is	NN	O	O
relatively	NN	O	O
resistant	NN	O	O
to	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

To	NN	O	O
further	NN	O	O
explore	NN	O	O
the	NN	O	O
link	NN	O	O
between	NN	O	O
the	NN	O	O
HPA	NN	O	O
and	NN	O	O
the	NN	O	O
immune	NN	O	O
system	NN	O	O
in	NN	O	O
relation	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
function	NN	O	O
,	NN	O	O
dose-response	NN	O	O
curves	NN	O	O
were	NN	O	O
obtained	NN	O	O
for	NN	O	O
Con	NN	O	O
A	NN	O	O
-and	NN	O	O
PHA	NN	O	B-protein
-induced	NN	O	O
T-cell	NN	O	O
mitogenesis	NN	O	O
,	NN	O	O
PWM	NN	O	B-protein
-generated	NN	O	O
B-cell	NN	O	O
mitogenesis	NN	O	O
and	NN	O	O
spontaneous	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
in	NN	O	O
13	NN	O	O
healthy	NN	O	O
controls	NN	O	O
.	NN	O	O

Glucocorticoid	NN	O	O
effects	NN	O	O
were	NN	O	O
assessed	NN	O	O
in	NN	O	O
vivo	NN	O	O
by	NN	O	O
depletion	NN	O	O
of	NN	O	O
endogenous	NN	O	O
glucocorticoids	NN	O	O
after	NN	O	O
oral	NN	O	O
administration	NN	O	O
of	NN	O	O
1.5	NN	O	O
g	NN	O	O
metyrapone	NN	O	O
(	NN	O	O
MET	NN	O	O
)	NN	O	O
and	NN	O	O
subsequent	NN	O	O
glucocorticoid	NN	O	O
replacement	NN	O	O
,	NN	O	O
and	NN	O	O
in	NN	O	O
vitro	NN	O	O
by	NN	O	O
incubation	NN	O	O
of	NN	O	O
the	NN	O	O
cells	NN	O	O
with	NN	O	O
different	NN	O	O
doses	NN	O	O
of	NN	O	O
dexamethasone	NN	O	O
(	NN	O	O
DEX	NN	O	O
)	NN	O	O
.	NN	O	O

There	NN	O	O
was	NN	O	O
a	NN	O	O
significant	NN	O	O
decrease	NN	O	O
in	NN	O	O
PWM	NN	O	B-protein
-induced	NN	O	O
B-cell	NN	O	O
mitogenesis	NN	O	O
and	NN	O	O
a	NN	O	O
more	NN	O	O
pronounced	NN	O	O
effect	NN	O	O
of	NN	O	O
DEX	NN	O	O
administered	NN	O	O
in	NN	O	O
vitro	NN	O	O
on	NN	O	O
spontaneous	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
after	NN	O	O
MET	NN	O	O
treatment	NN	O	O
when	NN	O	O
compared	NN	O	O
with	NN	O	O
the	NN	O	O
DEX	NN	O	O
plus	NN	O	O
MET	NN	O	O
pretreated	NN	O	O
condition	NN	O	O
in	NN	O	O
vivo	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
inhibition	NN	O	O
of	NN	O	O
spontaneous	NN	O	O
lymphocyte	NN	O	O
proliferation	NN	O	O
by	NN	O	O
glucocorticoids	NN	O	O
in	NN	O	O
vitro	NN	O	O
is	NN	O	O
related	NN	O	O
to	NN	O	O
glucocorticoid	NN	O	B-protein
receptor	NN	O	I-protein
function	NN	O	O
.	NN	O	O

The	NN	O	O
decrease	NN	O	O
in	NN	O	O
PWM	NN	O	B-protein
-generated	NN	O	O
B-cell	NN	O	O
proliferation	NN	O	O
following	NN	O	O
cortisol	NN	O	O
depletion	NN	O	O
by	NN	O	O
MET	NN	O	O
may	NN	O	O
be	NN	O	O
seen	NN	O	O
in	NN	O	O
connection	NN	O	O
with	NN	O	O
impaired	NN	O	O
glucocorticoid-mediated	NN	O	O
induction	NN	O	O
of	NN	O	O
interleukin-1	NN	O	B-protein
receptor	NN	O	I-protein
synthesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	O
specific	NN	O	O
differences	NN	O	O
visualized	NN	O	O
by	NN	O	O
fluorescence	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
pattern	NN	O	O
on	NN	O	O
interphase	NN	O	O
nuclei	NN	O	O
of	NN	O	O
different	NN	O	O
cell	NN	O	O
types	NN	O	O
.	NN	O	O

Application	NN	O	O
of	NN	O	O
a	NN	O	O
``	NN	O	O
formamide	NN	O	O
free	NN	O	O
''	NN	O	O
and	NN	O	O
thus	NN	O	O
``	NN	O	O
material	NN	O	O
preserving	NN	O	O
''	NN	O	O
in	NN	O	O
situ	NN	O	O
hybridization	NN	O	O
technique	NN	O	O
using	NN	O	O
the	NN	O	O
cDNA	NN	O	B-DNA
of	NN	O	O
the	NN	O	O
myf3	NN	O	B-DNA
gene	NN	O	I-DNA
revealed	NN	O	O
the	NN	O	O
following	NN	O	O
results	NN	O	O
:	NN	O	O
Human	NN	O	B-cell_type
rhabdomyosarcoma	NN	O	I-cell_type
cells	NN	O	I-cell_type
,	NN	O	O
characterized	NN	O	O
by	NN	O	O
a	NN	O	O
high	NN	O	O
expression	NN	O	O
of	NN	O	O
myf3	NN	O	B-DNA
show	NN	O	O
intensive	NN	O	O
hybridization	NN	O	O
signals	NN	O	O
in	NN	O	O
their	NN	O	O
interphase	NN	O	O
.	NN	O	O

RNase	NN	O	B-protein
treatment	NN	O	O
prior	NN	O	O
to	NN	O	O
hybridization	NN	O	O
considerably	NN	O	O
reduces	NN	O	O
the	NN	O	O
size	NN	O	O
of	NN	O	O
this	NN	O	O
signals	NN	O	O
.	NN	O	O

In	NN	O	O
comparison	NN	O	O
,	NN	O	O
isolated	NN	O	O
nuclei	NN	O	O
of	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
which	NN	O	O
no	NN	O	O
need	NN	O	O
for	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
this	NN	O	O
gene	NN	O	O
exists	NN	O	O
,	NN	O	O
show	NN	O	O
barely	NN	O	O
hybridization	NN	O	O
signals	NN	O	O
.	NN	O	O

Correspondingly	NN	O	O
,	NN	O	O
RNase	NN	O	B-protein
treatment	NN	O	O
had	NN	O	O
no	NN	O	O
effect	NN	O	O
on	NN	O	O
hybridization	NN	O	O
pattern	NN	O	O
at	NN	O	O
all	NN	O	O
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
an	NN	O	O
increased	NN	O	O
transcription	NN	O	O
efficiency	NN	O	O
of	NN	O	O
a	NN	O	O
cell	NN	O	O
type	NN	O	O
specific	NN	O	O
gene	NN	O	O
is	NN	O	O
accompanied	NN	O	O
by	NN	O	O
a	NN	O	O
higher	NN	O	O
hybridization	NN	O	O
accessibility	NN	O	O
in	NN	O	O
the	NN	O	O
corresponding	NN	O	O
cell	NN	O	O
nuclei	NN	O	O
.	NN	O	O

-DOCSTART-	O

Oncogenicity	NN	O	O
of	NN	O	O
human	NN	O	B-cell_line
papillomavirus-	NN	O	I-cell_line
or	NN	O	I-cell_line
adenovirus-transformed	NN	O	I-cell_line
cells	NN	O	I-cell_line
correlates	NN	O	O
with	NN	O	O
resistance	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
cells	NN	O	I-cell_type
.	NN	O	O

The	NN	O	O
reasons	NN	O	O
for	NN	O	O
the	NN	O	O
dissimilar	NN	O	O
oncogenicities	NN	O	O
of	NN	O	O
human	NN	O	O
adenoviruses	NN	O	O
and	NN	O	O
human	NN	O	O
papillomaviruses	NN	O	O
(	NN	O	O
HPV	NN	O	O
)	NN	O	O
in	NN	O	O
humans	NN	O	O
are	NN	O	O
unknown	NN	O	O
but	NN	O	O
may	NN	O	O
relate	NN	O	O
to	NN	O	O
differences	NN	O	O
in	NN	O	O
the	NN	O	O
capacities	NN	O	O
of	NN	O	O
the	NN	O	O
E1A	NN	O	B-protein
and	NN	O	O
E7	NN	O	B-protein
proteins	NN	O	I-protein
to	NN	O	O
target	NN	O	B-cell_type
cells	NN	O	I-cell_type
for	NN	O	O
rejection	NN	O	O
by	NN	O	O
the	NN	O	O
host	NN	O	B-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
cell	NN	O	I-cell_type
response	NN	O	O
.	NN	O	O

As	NN	O	O
one	NN	O	O
test	NN	O	O
of	NN	O	O
this	NN	O	O
hypothesis	NN	O	O
,	NN	O	O
we	NN	O	O
compared	NN	O	O
the	NN	O	O
abilities	NN	O	O
of	NN	O	O
E1A	NN	O	B-protein
-and	NN	O	O
E7	NN	O	B-protein
-expressing	NN	O	O
human	NN	O	B-cell_line
fibroblastic	NN	O	I-cell_line
or	NN	O	I-cell_line
keratinocyte-derived	NN	O	I-cell_line
human	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
be	NN	O	O
selectively	NN	O	O
killed	NN	O	O
by	NN	O	O
either	NN	O	O
unstimulated	NN	O	B-cell_type
or	NN	O	O
interferon	NN	O	B-cell_line
(	NN	O	I-cell_line
IFN	NN	O	I-cell_line
)	NN	O	I-cell_line
-activated	NN	O	I-cell_line
NK	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Cells	NN	O	O
expressing	NN	O	O
the	NN	O	O
E1A	NN	O	B-protein
oncoprotein	NN	O	I-protein
were	NN	O	O
selectively	NN	O	O
killed	NN	O	O
by	NN	O	O
unstimulated	NN	O	B-cell_line
NK	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
while	NN	O	O
the	NN	O	O
same	NN	O	O
parental	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
expressing	NN	O	O
the	NN	O	O
HPV	NN	O	O
type	NN	O	O
16	NN	O	O
(	NN	O	O
HPV-16	NN	O	O
)	NN	O	O
or	NN	O	O
HPV-18	NN	O	O
E7	NN	O	B-protein
oncoprotein	NN	O	I-protein
were	NN	O	O
resistant	NN	O	O
to	NN	O	O
NK	NN	O	O
cell	NN	O	O
lysis	NN	O	O
.	NN	O	O

The	NN	O	O
ability	NN	O	O
of	NN	O	O
IFN-activated	NN	O	B-cell_line
NK	NN	O	I-cell_line
cells	NN	O	I-cell_line
to	NN	O	O
selectively	NN	O	O
kill	NN	O	O
virally	NN	O	O
transformed	NN	O	O
cells	NN	O	O
depends	NN	O	O
on	NN	O	O
IFN	NN	O	B-protein
's	NN	O	O
ability	NN	O	O
to	NN	O	O
induce	NN	O	O
resistance	NN	O	O
to	NN	O	O
NK	NN	O	O
cell	NN	O	O
lysis	NN	O	O
in	NN	O	O
normal	NN	O	O
(	NN	O	O
i.e.	NN	O	O
,	NN	O	O
non-viral	NN	O	B-cell_line
oncogene-expressing	NN	O	I-cell_line
)	NN	O	O
but	NN	O	O
not	NN	O	O
virally	NN	O	B-cell_line
transformed	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

E1A	NN	O	B-protein
blocked	NN	O	O
IFN	NN	O	B-protein
's	NN	O	O
induction	NN	O	O
of	NN	O	O
cytolytic	NN	O	O
resistance	NN	O	O
,	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
selective	NN	O	O
lysis	NN	O	O
of	NN	O	O
adenovirus-transformed	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
IFN-activated	NN	O	B-cell_line
NK	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

The	NN	O	O
extent	NN	O	O
of	NN	O	O
IFN	NN	O	B-protein
-induced	NN	O	O
NK	NN	O	O
cell	NN	O	O
killing	NN	O	O
of	NN	O	O
E1A-expressing	NN	O	B-cell_line
cells	NN	O	I-cell_line
was	NN	O	O
proportional	NN	O	O
to	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
E1A	NN	O	B-protein
expression	NN	O	O
and	NN	O	O
correlated	NN	O	O
with	NN	O	O
the	NN	O	O
ability	NN	O	O
of	NN	O	O
E1A	NN	O	B-protein
to	NN	O	O
block	NN	O	O
IFN	NN	O	B-protein
-stimulated	NN	O	O
gene	NN	O	O
expression	NN	O	O
in	NN	O	O
target	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
E7	NN	O	B-protein
blocked	NN	O	O
neither	NN	O	O
IFN	NN	O	B-protein
-stimulated	NN	O	O
gene	NN	O	O
expression	NN	O	O
nor	NN	O	O
IFN	NN	O	B-protein
's	NN	O	O
induction	NN	O	O
of	NN	O	O
cytolytic	NN	O	O
resistance	NN	O	O
,	NN	O	O
thereby	NN	O	O
precluding	NN	O	O
the	NN	O	O
selective	NN	O	O
lysis	NN	O	O
of	NN	O	O
HPV-transformed	NN	O	B-cell_line
cells	NN	O	I-cell_line
by	NN	O	O
IFN-activated	NN	O	B-cell_line
NK	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

In	NN	O	O
conclusion	NN	O	O
,	NN	O	O
E1A	NN	O	B-protein
expression	NN	O	O
marks	NN	O	O
cells	NN	O	O
for	NN	O	O
destruction	NN	O	O
by	NN	O	O
the	NN	O	O
host	NN	O	O
NK	NN	O	B-cell_type
cell	NN	O	I-cell_type
response	NN	O	O
,	NN	O	O
whereas	NN	O	O
the	NN	O	O
E7	NN	O	B-protein
oncoprotein	NN	O	I-protein
lacks	NN	O	O
this	NN	O	O
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Regulation	NN	O	O
of	NN	O	O
IkB	NN	O	B-protein
alpha	NN	O	I-protein
phosphorylation	NN	O	O
by	NN	O	O
PKC-	NN	O	O
and	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
signal	NN	O	O
transduction	NN	O	O
pathways	NN	O	O
.	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
-dependent	NN	O	I-protein
phosphatase	NN	O	I-protein
calcineurin	NN	O	B-protein
,	NN	O	O
a	NN	O	O
target	NN	O	O
of	NN	O	O
FK506	NN	O	O
and	NN	O	O
CsA	NN	O	O
,	NN	O	O
synergizes	NN	O	O
with	NN	O	O
PKC	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
(	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
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
have	NN	O	O
investigated	NN	O	O
whether	NN	O	O
this	NN	O	O
synergy	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
and	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
these	NN	O	O
two	NN	O	O
pathways	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
.	NN	O	O

While	NN	O	O
this	NN	O	O
synergy	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
,	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
U937	NN	O	I-cell_line
calcineurin	NN	O	B-protein
is	NN	O	O
also	NN	O	O
sufficient	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

Having	NN	O	O
previously	NN	O	O
shown	NN	O	O
that	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-	NN	O	O
and	NN	O	O
PKC-dependent	NN	O	O
pathways	NN	O	O
synergize	NN	O	O
by	NN	O	O
accelerating	NN	O	O
the	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkB	NN	O	B-protein
alpha	NN	O	I-protein
,	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
IkB	NN	O	B-protein
alpha	NN	O	I-protein
phosphorylation	NN	O	O
.	NN	O	O

While	NN	O	O
PKC	NN	O	B-protein
-dependent	NN	O	O
pathways	NN	O	O
sequentially	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
in	NN	O	O
an	NN	O	O
incomplete	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkB	NN	O	B-protein
alpha	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
,	NN	O	O
co-activation	NN	O	O
of	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
pathways	NN	O	O
accelerates	NN	O	O
the	NN	O	O
rate	NN	O	O
of	NN	O	O
IkB	NN	O	B-protein
alpha	NN	O	I-protein
phosphorylation	NN	O	O
and	NN	O	O
results	NN	O	O
in	NN	O	O
its	NN	O	O
complete	NN	O	O
degradation	NN	O	O
.	NN	O	O

Activation	NN	O	O
of	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
pathways	NN	O	O
alone	NN	O	O
do	NN	O	O
not	NN	O	O
result	NN	O	O
in	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
and/or	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkB	NN	O	B-protein
alpha	NN	O	I-protein
in	NN	O	O
Jurkat	NN	O	B-cell_line
T	NN	O	I-cell_line
or	NN	O	O
in	NN	O	O
U937	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	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
selective	NN	O	O
PKC	NN	O	B-protein
inhibitor	NN	O	O
GF109203X	NN	O	O
abrogates	NN	O	O
the	NN	O	O
PMA-induced	NN	O	O
IkB	NN	O	B-protein
alpha	NN	O	I-protein
phosphorylation/degradation	NN	O	O
irrespective	NN	O	O
of	NN	O	O
activation	NN	O	O
of	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
pathways	NN	O	O
,	NN	O	O
but	NN	O	O
not	NN	O	O
the	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
of	NN	O	O
IkB	NN	O	B-protein
alpha	NN	O	I-protein
induced	NN	O	O
by	NN	O	O
TNF-alpha	NN	O	B-protein
,	NN	O	O
a	NN	O	O
PKC	NN	O	B-protein
-independent	NN	O	O
stimulus	NN	O	O
.	NN	O	O

Contrary	NN	O	O
to	NN	O	O
the	NN	O	O
interaction	NN	O	O
with	NN	O	O
PKC	NN	O	B-protein
,	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
pathways	NN	O	O
synergize	NN	O	O
with	NN	O	O
TNF-alpha	NN	O	B-protein
not	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
IkB	NN	O	B-protein
alpha	NN	O	I-protein
phosphorylation	NN	O	O
,	NN	O	O
but	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
its	NN	O	O
degradation	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
Ca	NN	O	O
(	NN	O	O
2+	NN	O	O
)	NN	O	O
-dependent	NN	O	O
pathways	NN	O	O
,	NN	O	O
including	NN	O	O
the	NN	O	O
phosphatase	NN	O	B-protein
calcineurin	NN	O	B-protein
,	NN	O	O
participate	NN	O	O
in	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
in	NN	O	O
a	NN	O	O
cell	NN	O	O
specific	NN	O	O
fashion	NN	O	O
and	NN	O	O
synergize	NN	O	O
with	NN	O	O
PKC	NN	O	B-protein
-dependent	NN	O	O
and	NN	O	O
-independent	NN	O	O
pathways	NN	O	O
at	NN	O	O
the	NN	O	O
level	NN	O	O
of	NN	O	O
IkB	NN	O	B-protein
alpha	NN	O	I-protein
phosphorylation	NN	O	O
and	NN	O	O
degradation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Vitamin	NN	O	O
E	NN	O	O
therapy	NN	O	O
of	NN	O	O
acute	NN	O	O
CCl4-induced	NN	O	O
hepatic	NN	O	O
injury	NN	O	O
in	NN	O	O
mice	NN	O	O
is	NN	O	O
associated	NN	O	O
with	NN	O	O
inhibition	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
binding	NN	O	O
.	NN	O	O

Oxidative	NN	O	O
stress	NN	O	O
,	NN	O	O
with	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediate	NN	O	O
formation	NN	O	O
,	NN	O	O
may	NN	O	O
represent	NN	O	O
a	NN	O	O
common	NN	O	O
mechanism	NN	O	O
by	NN	O	O
which	NN	O	O
liver	NN	O	O
injury	NN	O	O
is	NN	O	O
induced	NN	O	O
by	NN	O	O
diverse	NN	O	O
etiologies	NN	O	O
.	NN	O	O

Oxidative	NN	O	O
stress	NN	O	O
enhances	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
activity	NN	O	O
,	NN	O	O
and	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
shown	NN	O	O
to	NN	O	O
enhance	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
cytotoxic	NN	O	B-protein
cytokines	NN	O	I-protein
.	NN	O	O

Acute	NN	O	O
hepatic	NN	O	O
injury	NN	O	O
caused	NN	O	O
by	NN	O	O
reactive	NN	O	O
oxygen	NN	O	O
intermediate	NN	O	O
production	NN	O	O
was	NN	O	O
induced	NN	O	O
by	NN	O	O
an	NN	O	O
intraperitoneal	NN	O	O
injection	NN	O	O
of	NN	O	O
CCl4	NN	O	O
in	NN	O	O
mice	NN	O	O
.	NN	O	O

This	NN	O	O
injury	NN	O	O
was	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
intravenous	NN	O	O
pretreatment	NN	O	O
of	NN	O	O
the	NN	O	O
mice	NN	O	O
with	NN	O	O
a	NN	O	O
water-soluble	NN	O	O
emulsion	NN	O	O
of	NN	O	O
alpha-tocopherol	NN	O	O
.	NN	O	O

Alpha-tocopherol	NN	O	O
treatment	NN	O	O
of	NN	O	O
the	NN	O	O
mice	NN	O	O
given	NN	O	O
the	NN	O	O
CCl4	NN	O	O
also	NN	O	O
reduced	NN	O	O
the	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
to	NN	O	O
levels	NN	O	O
approaching	NN	O	O
those	NN	O	O
found	NN	O	O
in	NN	O	O
normal	NN	O	O
mice	NN	O	O
.	NN	O	O

In	NN	O	O
vitro	NN	O	O
treatment	NN	O	O
of	NN	O	O
a	NN	O	O
monocyte/macrophage	NN	O	O
cell	NN	O	O
line	NN	O	O
with	NN	O	O
CCl4	NN	O	O
led	NN	O	O
to	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
an	NN	O	O
increase	NN	O	O
in	NN	O	O
tumor	NN	O	B-RNA
necrosis	NN	O	I-RNA
factor-alpha	NN	O	I-RNA
(	NN	O	I-RNA
TNF-alpha	NN	O	I-RNA
)	NN	O	I-RNA
messenger	NN	O	I-RNA
RNA	NN	O	I-RNA
levels	NN	O	O
.	NN	O	O

Liver	NN	O	O
specimens	NN	O	O
taken	NN	O	O
from	NN	O	O
patients	NN	O	O
with	NN	O	O
acute	NN	O	O
fulminant	NN	O	O
hepatitis	NN	O	O
had	NN	O	O
markedly	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
comparison	NN	O	O
with	NN	O	O
the	NN	O	O
binding	NN	O	O
of	NN	O	O
normal	NN	O	O
livers	NN	O	O
.	NN	O	O

These	NN	O	O
data	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
abolishing	NN	O	O
acute	NN	O	O
hepatic	NN	O	O
injury	NN	O	O
with	NN	O	O
alpha-tocopherol	NN	O	O
,	NN	O	O
a	NN	O	O
free	NN	O	O
radical	NN	O	O
scavenger	NN	O	O
,	NN	O	O
also	NN	O	O
eliminated	NN	O	O
increased	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
binding	NN	O	O
.	NN	O	O

It	NN	O	O
is	NN	O	O
tempting	NN	O	O
to	NN	O	O
speculate	NN	O	O
that	NN	O	O
enhanced	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
expression	NN	O	O
caused	NN	O	O
by	NN	O	O
free	NN	O	O
radical	NN	O	O
production/oxidative	NN	O	O
stress	NN	O	O
may	NN	O	O
modulate	NN	O	O
liver	NN	O	O
injury	NN	O	O
,	NN	O	O
perhaps	NN	O	O
through	NN	O	O
an	NN	O	O
effect	NN	O	O
on	NN	O	O
cytotoxic	NN	O	B-protein
cytokine	NN	O	I-protein
synthesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

Immunosuppression	NN	O	O
by	NN	O	O
glucocorticoids	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
activity	NN	O	O
through	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
synthesis	NN	O	O
[	NN	O	O
see	NN	O	O
comments	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
anti-inflammatory	NN	O	O
and	NN	O	O
immunosuppressive	NN	O	O
agents	NN	O	O
.	NN	O	O

They	NN	O	O
inhibit	NN	O	O
synthesis	NN	O	O
of	NN	O	O
almost	NN	O	O
all	NN	O	O
known	NN	O	O
cytokines	NN	O	B-protein
and	NN	O	O
of	NN	O	O
several	NN	O	O
cell	NN	O	B-protein
surface	NN	O	I-protein
molecules	NN	O	I-protein
required	NN	O	O
for	NN	O	O
immune	NN	O	O
function	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
mechanism	NN	O	O
underlying	NN	O	O
this	NN	O	O
activity	NN	O	O
has	NN	O	O
been	NN	O	O
unclear.	NN	O	O
Here	NN	O	O
it	NN	O	O
is	NN	O	O
shown	NN	O	O
that	NN	O	O
glucocorticoids	NN	O	O
are	NN	O	O
potent	NN	O	O
inhibitors	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
in	NN	O	O
mice	NN	O	O
and	NN	O	O
cultured	NN	O	B-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

This	NN	O	O
inhibition	NN	O	O
is	NN	O	O
mediated	NN	O	O
by	NN	O	O
induction	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	O
inhibitory	NN	O	O
protein	NN	O	O
,	NN	O	O
which	NN	O	O
traps	NN	O	O
activated	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
in	NN	O	O
inactive	NN	O	B-protein
cytoplasmic	NN	O	I-protein
complexes	NN	O	I-protein
.	NN	O	O

Because	NN	O	O
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
activates	NN	O	O
many	NN	O	O
immunoregulatory	NN	O	B-DNA
genes	NN	O	I-DNA
in	NN	O	O
response	NN	O	O
to	NN	O	O
pro-inflammatory	NN	O	O
stimuli	NN	O	O
,	NN	O	O
the	NN	O	O
inhibition	NN	O	O
of	NN	O	O
its	NN	O	O
activity	NN	O	O
can	NN	O	O
be	NN	O	O
a	NN	O	O
major	NN	O	O
component	NN	O	O
of	NN	O	O
the	NN	O	O
anti-inflammatory	NN	O	O
activity	NN	O	O
of	NN	O	O
glucocorticoids	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcriptional	NN	O	O
repression	NN	O	O
of	NN	O	O
the	NN	O	O
interleukin-2	NN	O	B-DNA
gene	NN	O	I-DNA
by	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
:	NN	O	O
direct	NN	O	O
inhibition	NN	O	O
of	NN	O	O
NFATp/AP-1	NN	O	B-protein
complex	NN	O	I-protein
formation	NN	O	O
by	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
hormone	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

T-lymphocyte	NN	O	O
proliferation	NN	O	O
is	NN	O	O
suppressed	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
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
the	NN	O	O
active	NN	O	O
metabolite	NN	O	O
of	NN	O	O
vitamin	NN	O	O
D3	NN	O	O
,	NN	O	O
and	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
interleukin	NN	O	O
2	NN	O	O
(	NN	O	O
IL-2	NN	O	O
)	NN	O	O
,	NN	O	O
gamma	NN	O	O
interferon	NN	O	O
,	NN	O	O
and	NN	O	O
granulocyte-macrophage	NN	O	O
colony-stimulating	NN	O	O
factor	NN	O	O
mRNA	NN	O	O
levels	NN	O	O
.	NN	O	O

We	NN	O	O
report	NN	O	O
here	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-mediated	NN	O	O
repression	NN	O	O
in	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
is	NN	O	O
cycloheximide	NN	O	O
resistant	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
it	NN	O	O
is	NN	O	O
a	NN	O	O
direct	NN	O	O
transcriptional	NN	O	O
repressive	NN	O	O
effect	NN	O	O
on	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
by	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
VDR	NN	O	B-protein
)	NN	O	O
.	NN	O	O

We	NN	O	O
therefore	NN	O	O
examined	NN	O	O
vitamin	NN	O	O
D3-mediated	NN	O	O
repression	NN	O	O
of	NN	O	O
activated	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
by	NN	O	O
cotransfecting	NN	O	O
Jurkat	NN	O	B-cell_line
cells	NN	O	I-cell_line
with	NN	O	O
IL-2	NN	O	B-DNA
promoter/reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
and	NN	O	O
a	NN	O	O
VDR	NN	O	B-DNA
overexpression	NN	O	I-DNA
vector	NN	O	I-DNA
and	NN	O	O
by	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

We	NN	O	O
delineated	NN	O	O
an	NN	O	O
element	NN	O	O
conferring	NN	O	O
both	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
the	NN	O	O
receptor	NN	O	O
in	NN	O	O
vitro	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-mediated	NN	O	O
repression	NN	O	O
in	NN	O	O
vivo	NN	O	O
to	NN	O	O
a	NN	O	O
short	NN	O	O
40-bp	NN	O	B-DNA
region	NN	O	I-DNA
encompassing	NN	O	O
an	NN	O	O
important	NN	O	O
positive	NN	O	B-DNA
regulatory	NN	O	I-DNA
element	NN	O	I-DNA
,	NN	O	O
NF-AT-1	NN	O	B-DNA
,	NN	O	O
which	NN	O	O
is	NN	O	O
bound	NN	O	O
by	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
NFATp	NN	O	B-protein
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
by	NN	O	O
AP-1	NN	O	B-protein
.	NN	O	O

VDR	NN	O	B-protein
DNA-binding	NN	O	I-protein
mutants	NN	O	I-protein
were	NN	O	O
unable	NN	O	O
to	NN	O	O
either	NN	O	O
bind	NN	O	O
to	NN	O	O
this	NN	O	O
element	NN	O	O
in	NN	O	O
vitro	NN	O	O
or	NN	O	O
repress	NN	O	O
in	NN	O	O
vivo	NN	O	O
;	NN	O	O
the	NN	O	O
VDR	NN	O	B-protein
DNA-binding	NN	O	I-protein
domain	NN	O	I-protein
alone	NN	O	O
,	NN	O	O
however	NN	O	O
,	NN	O	O
bound	NN	O	O
the	NN	O	O
element	NN	O	O
but	NN	O	O
also	NN	O	O
could	NN	O	O
not	NN	O	O
repress	NN	O	O
IL-2	NN	O	B-protein
expression	NN	O	O
.	NN	O	O

These	NN	O	O
results	NN	O	O
indicate	NN	O	O
that	NN	O	O
DNA	NN	O	O
binding	NN	O	O
by	NN	O	O
VDR	NN	O	B-protein
is	NN	O	O
necessary	NN	O	O
but	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
mediate	NN	O	O
IL-2	NN	O	B-protein
repression	NN	O	O
.	NN	O	O

By	NN	O	O
combining	NN	O	O
partially	NN	O	B-protein
purified	NN	O	I-protein
proteins	NN	O	I-protein
in	NN	O	O
vitro	NN	O	O
,	NN	O	O
we	NN	O	O
observed	NN	O	O
the	NN	O	O
loss	NN	O	O
of	NN	O	O
the	NN	O	O
bound	NN	O	O
NFATp/AP-1-DNA	NN	O	B-protein
complex	NN	O	I-protein
upon	NN	O	O
inclusion	NN	O	O
of	NN	O	O
VDR	NN	O	B-protein
or	NN	O	O
VDR-retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
.	NN	O	O

Order	NN	O	O
of	NN	O	O
addition	NN	O	O
and	NN	O	O
off-rate	NN	O	O
experiments	NN	O	O
indicate	NN	O	O
that	NN	O	O
the	NN	O	O
VDR-retinoid	NN	O	B-protein
X	NN	O	I-protein
receptor	NN	O	I-protein
heterodimer	NN	O	I-protein
blocks	NN	O	O
NFATp/AP-1	NN	O	B-protein
complex	NN	O	I-protein
formation	NN	O	O
and	NN	O	O
then	NN	O	O
stably	NN	O	O
associates	NN	O	O
with	NN	O	O
the	NN	O	O
NF-AT-1	NN	O	B-DNA
element	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
direct	NN	O	O
inhibition	NN	O	O
by	NN	O	O
a	NN	O	O
nuclear	NN	O	O
hormone	NN	O	O
receptor	NN	O	O
of	NN	O	O
transcriptional	NN	O	B-protein
activators	NN	O	I-protein
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
may	NN	O	O
provide	NN	O	O
a	NN	O	O
mechanistic	NN	O	O
explanation	NN	O	O
of	NN	O	O
how	NN	O	O
vitamin	NN	O	O
derivatives	NN	O	O
can	NN	O	O
act	NN	O	O
as	NN	O	O
potent	NN	O	O
immunosuppressive	NN	O	O
agents	NN	O	O
.	NN	O	O

-DOCSTART-	O

Tissue-specific	NN	O	O
regulation	NN	O	O
of	NN	O	O
the	NN	O	O
rabbit	NN	O	B-DNA
15-lipoxygenase	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-DNA
silencer	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
15-lipoxygenase	NN	O	B-DNA
(	NN	O	I-DNA
lox	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
a	NN	O	O
tissue-specific	NN	O	O
manner	NN	O	O
,	NN	O	O
predominantly	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
also	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
and	NN	O	O
eosinophils	NN	O	B-cell_type
.	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
the	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
DNA	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
15-lox	NN	O	B-DNA
gene	NN	O	I-DNA
contains	NN	O	O
sequences	NN	O	O
which	NN	O	O
down-regulate	NN	O	O
its	NN	O	O
activity	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
non-erythroid	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
but	NN	O	O
not	NN	O	O
in	NN	O	O
two	NN	O	O
erythroid	NN	O	B-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
has	NN	O	O
characteristics	NN	O	O
of	NN	O	O
a	NN	O	O
transcriptional	NN	O	B-DNA
'silencer	NN	O	I-DNA
'	NN	O	I-DNA
since	NN	O	O
it	NN	O	O
functions	NN	O	O
in	NN	O	O
both	NN	O	O
orientations	NN	O	O
.	NN	O	O

The	NN	O	O
main	NN	O	O
activity	NN	O	O
of	NN	O	O
the	NN	O	O
silencer	NN	O	B-DNA
has	NN	O	O
been	NN	O	O
mapped	NN	O	O
to	NN	O	O
the	NN	O	O
first	NN	O	O
900	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
DNA	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
contains	NN	O	O
nine	NN	O	O
binding	NN	O	O
sites	NN	O	O
for	NN	O	O
a	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
present	NN	O	O
in	NN	O	O
non-	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
but	NN	O	O
not	NN	O	O
in	NN	O	O
erythroid	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

These	NN	O	O
binding	NN	O	O
sites	NN	O	O
have	NN	O	O
similar	NN	O	O
sequences	NN	O	O
and	NN	O	O
multiple	NN	O	O
copies	NN	O	O
of	NN	O	O
the	NN	O	O
binding	NN	O	B-DNA
sites	NN	O	I-DNA
confer	NN	O	O
tissue-specific	NN	O	O
down-regulation	NN	O	O
when	NN	O	O
attached	NN	O	O
to	NN	O	O
a	NN	O	O
minimal	NN	O	B-DNA
lox	NN	O	I-DNA
promoter	NN	O	I-DNA
fragment	NN	O	I-DNA
.	NN	O	O

The	NN	O	O
5	NN	O	B-DNA
'	NN	O	I-DNA
flanking	NN	O	I-DNA
DNA	NN	O	I-DNA
also	NN	O	O
contains	NN	O	O
a	NN	O	O
cluster	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
the	NN	O	O
GATA	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

-DOCSTART-	O

Phosphorylation	NN	O	O
of	NN	O	O
the	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
NFATp	NN	O	B-protein
inhibits	NN	O	O
its	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
in	NN	O	O
cyclosporin	NN	O	B-cell_line
A-treated	NN	O	I-cell_line
human	NN	O	I-cell_line
B	NN	O	I-cell_line
and	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

Cyclosporin	NN	O	O
A	NN	O	O
(	NN	O	O
CsA	NN	O	O
)	NN	O	O
exerts	NN	O	O
its	NN	O	O
immunosuppressive	NN	O	O
effect	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
nuclear	NN	O	B-protein
factor	NN	O	I-protein
of	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
(	NN	O	O
NFAT	NN	O	B-protein
)	NN	O	O
,	NN	O	O
thus	NN	O	O
preventing	NN	O	O
transcriptional	NN	O	O
induction	NN	O	O
of	NN	O	O
several	NN	O	O
cytokine	NN	O	O
genes	NN	O	O
.	NN	O	O

This	NN	O	O
effect	NN	O	O
is	NN	O	O
thought	NN	O	O
to	NN	O	O
be	NN	O	O
largely	NN	O	O
mediated	NN	O	O
through	NN	O	O
inactivation	NN	O	O
of	NN	O	O
the	NN	O	O
phosphatase	NN	O	O
calcineurin	NN	O	B-protein
,	NN	O	O
which	NN	O	O
in	NN	O	O
turn	NN	O	O
inhibits	NN	O	O
translocation	NN	O	O
of	NN	O	O
an	NN	O	O
NFAT	NN	O	B-protein
component	NN	O	O
to	NN	O	O
the	NN	O	O
nucleus	NN	O	O
.	NN	O	O

Here	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
CsA	NN	O	O
treatment	NN	O	O
of	NN	O	O
Raji	NN	O	B-cell_line
B	NN	O	I-cell_line
and	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
yields	NN	O	O
a	NN	O	O
phosphorylated	NN	O	O
form	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
that	NN	O	O
is	NN	O	O
inhibited	NN	O	O
in	NN	O	O
DNA-binding	NN	O	O
and	NN	O	O
in	NN	O	O
its	NN	O	O
ability	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
with	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
.	NN	O	O

Immunoblot	NN	O	O
analyses	NN	O	O
and	NN	O	O
metabolic	NN	O	O
labeling	NN	O	O
with	NN	O	O
[	NN	O	O
32P	NN	O	O
]	NN	O	O
orthophosphate	NN	O	O
show	NN	O	O
that	NN	O	O
CsA	NN	O	O
alters	NN	O	O
NFATp	NN	O	B-protein
migration	NN	O	O
on	NN	O	O
SDS-polyacrylamide	NN	O	O
gel	NN	O	O
electrophoresis	NN	O	O
by	NN	O	O
increasing	NN	O	O
its	NN	O	O
phosphorylation	NN	O	O
level	NN	O	O
without	NN	O	O
affecting	NN	O	O
subcellular	NN	O	O
distribution	NN	O	O
.	NN	O	O

Dephosphorylation	NN	O	O
by	NN	O	O
in	NN	O	O
vitro	NN	O	O
treatment	NN	O	O
with	NN	O	O
calcineurin	NN	O	B-protein
or	NN	O	O
alkaline	NN	O	B-protein
phosphatase	NN	O	I-protein
restores	NN	O	O
NFATp	NN	O	B-protein
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
and	NN	O	O
its	NN	O	O
ability	NN	O	O
to	NN	O	O
reconstitute	NN	O	O
an	NN	O	O
NFAT	NN	O	B-protein
complex	NN	O	I-protein
with	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
proteins	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
new	NN	O	O
mechanism	NN	O	O
for	NN	O	O
CsA-sensitive	NN	O	O
regulation	NN	O	O
of	NN	O	O
NFATp	NN	O	B-protein
in	NN	O	O
which	NN	O	O
dephosphorylation	NN	O	O
is	NN	O	O
critical	NN	O	O
for	NN	O	O
DNA	NN	O	O
binding	NN	O	O
.	NN	O	O

-DOCSTART-	O

Transcription	NN	O	B-protein
factors	NN	O	I-protein
as	NN	O	O
targets	NN	O	O
for	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

We	NN	O	O
previously	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
a	NN	O	O
requirement	NN	O	O
for	NN	O	O
oxidative	NN	O	O
events	NN	O	O
during	NN	O	O
cell	NN	O	O
cycle	NN	O	O
entry	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
have	NN	O	O
hypothesised	NN	O	O
that	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
intracellular	NN	O	O
signalling	NN	O	O
agents	NN	O	O
during	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
.	NN	O	O

In	NN	O	O
the	NN	O	O
current	NN	O	O
study	NN	O	O
,	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
antioxidant	NN	O	O
activity	NN	O	O
,	NN	O	O
has	NN	O	O
been	NN	O	O
used	NN	O	O
to	NN	O	O
further	NN	O	O
investigate	NN	O	O
the	NN	O	O
role	NN	O	O
of	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

Treatment	NN	O	O
of	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
lymphocytes	NN	O	I-cell_type
with	NN	O	O
cysteamine	NN	O	O
in	NN	O	O
vitro	NN	O	O
was	NN	O	O
found	NN	O	O
to	NN	O	O
inhibit	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
with	NN	O	O
essentially	NN	O	O
complete	NN	O	O
inhibition	NN	O	O
occurring	NN	O	O
at	NN	O	O
a	NN	O	O
dose	NN	O	O
of	NN	O	O
400	NN	O	O
microM	NN	O	O
.	NN	O	O

This	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
was	NN	O	O
limited	NN	O	O
to	NN	O	O
the	NN	O	O
first	NN	O	O
2	NN	O	O
h	NN	O	O
after	NN	O	O
mitogenic	NN	O	O
activation	NN	O	O
,	NN	O	O
localizing	NN	O	O
the	NN	O	O
time-frame	NN	O	O
of	NN	O	O
action	NN	O	O
of	NN	O	O
cysteamine	NN	O	O
to	NN	O	O
within	NN	O	O
the	NN	O	O
commitment	NN	O	O
period	NN	O	O
.	NN	O	O

It	NN	O	O
therefore	NN	O	O
was	NN	O	O
of	NN	O	O
interest	NN	O	O
to	NN	O	O
establish	NN	O	O
which	NN	O	O
,	NN	O	O
if	NN	O	O
any	NN	O	O
,	NN	O	O
commitment	NN	O	O
events	NN	O	O
were	NN	O	O
affected	NN	O	O
by	NN	O	O
oxidative	NN	O	O
signalling	NN	O	O
during	NN	O	O
cell	NN	O	O
cycle	NN	O	O
entry	NN	O	O
.	NN	O	O

Taking	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
gene	NN	O	I-DNA
as	NN	O	O
a	NN	O	O
candidate	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
cysteamine	NN	O	O
treatment	NN	O	O
on	NN	O	O
early	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
during	NN	O	O
lymphocyte	NN	O	O
activation	NN	O	O
,	NN	O	O
and	NN	O	O
on	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
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
and	NN	O	O
Oct1	NN	O	B-protein
,	NN	O	O
whose	NN	O	O
functions	NN	O	O
are	NN	O	O
required	NN	O	O
for	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-protein
mRNA	NN	O	O
.	NN	O	O

Cysteamine	NN	O	O
treatment	NN	O	O
inhibited	NN	O	O
both	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
IL-2	NN	O	B-RNA
mRNA	NN	O	I-RNA
and	NN	O	O
secretion	NN	O	O
of	NN	O	O
IL-2	NN	O	B-protein
into	NN	O	O
the	NN	O	O
culture	NN	O	O
medium	NN	O	O
.	NN	O	O

The	NN	O	O
inhibitory	NN	O	O
effect	NN	O	O
of	NN	O	O
cysteamine	NN	O	O
may	NN	O	O
be	NN	O	O
mediated	NN	O	O
at	NN	O	O
least	NN	O	O
in	NN	O	O
part	NN	O	O
by	NN	O	O
an	NN	O	O
effect	NN	O	O
on	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
function	NN	O	O
,	NN	O	O
as	NN	O	O
the	NN	O	O
DNA	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
NF-kappa	NN	O	B-protein
B	NN	O	I-protein
extracted	NN	O	O
from	NN	O	O
mitogen-stimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
were	NN	O	O
significantly	NN	O	O
inhibited	NN	O	O
by	NN	O	O
cysteamine	NN	O	O
treatment	NN	O	O
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
Oct1	NN	O	O
and	NN	O	O
NF-AT	NN	O	O
DNA	NN	O	O
binding	NN	O	O
activity	NN	O	O
were	NN	O	O
not	NN	O	O
affected	NN	O	O
by	NN	O	O
cysteamine	NN	O	O
treatment	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
oxidative	NN	O	O
signalling	NN	O	O
processes	NN	O	O
operate	NN	O	O
in	NN	O	O
a	NN	O	O
selective	NN	O	O
manner	NN	O	O
.	NN	O	O

The	NN	O	O
identification	NN	O	O
of	NN	O	O
regulatory	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
such	NN	O	O
as	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
,	NN	O	O
as	NN	O	O
molecular	NN	O	O
targets	NN	O	O
for	NN	O	O
oxidative	NN	O	O
signalling	NN	O	O
provides	NN	O	O
further	NN	O	O
evidence	NN	O	O
to	NN	O	O
implicate	NN	O	O
oxidative	NN	O	O
signalling	NN	O	O
as	NN	O	O
being	NN	O	O
intimately	NN	O	O
involved	NN	O	O
in	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
in	NN	O	O
T	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

-DOCSTART-	O

Sex	NN	O	O
and	NN	O	O
age	NN	O	O
distribution	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
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
human	NN	O	O
subjects	NN	O	O
.	NN	O	O

Specific	NN	O	O
receptors	NN	O	O
for	NN	O	O
1	NN	O	O
,	NN	O	O
25	NN	O	O
Dihydroxyvitamin	NN	O	O
D3	NN	O	O
have	NN	O	O
been	NN	O	O
described	NN	O	O
in	NN	O	O
human	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
tried	NN	O	O
to	NN	O	O
find	NN	O	O
out	NN	O	O
whether	NN	O	O
these	NN	O	O
receptors	NN	O	O
could	NN	O	O
show	NN	O	O
any	NN	O	O
difference	NN	O	O
in	NN	O	O
sex	NN	O	O
or	NN	O	O
age	NN	O	O
distribution	NN	O	O
.	NN	O	O

Twenty	NN	O	O
two	NN	O	O
healthy	NN	O	O
men	NN	O	O
aged	NN	O	O
21-66	NN	O	O
yr	NN	O	O
(	NN	O	O
mean	NN	O	O
+/-	NN	O	O
SD	NN	O	O
41.0	NN	O	O
+/-	NN	O	O
13.6	NN	O	O
)	NN	O	O
and	NN	O	O
nineteen	NN	O	O
healthy	NN	O	O
women	NN	O	O
aged	NN	O	O
22-60	NN	O	O
yr	NN	O	O
(	NN	O	O
38.9	NN	O	O
+/-	NN	O	O
13.9	NN	O	O
)	NN	O	O
have	NN	O	O
been	NN	O	O
studied	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
sexes	NN	O	O
(	NN	O	O
1.35	NN	O	O
+/-	NN	O	O
0.70	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
in	NN	O	O
males	NN	O	O
,	NN	O	O
1.13	NN	O	O
+/-	NN	O	O
0.66	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
in	NN	O	O
females	NN	O	O
)	NN	O	O
,	NN	O	O
but	NN	O	O
the	NN	O	O
concentration	NN	O	O
of	NN	O	O
binding	NN	O	O
sites	NN	O	O
(	NN	O	O
Nmax	NN	O	O
)	NN	O	O
was	NN	O	O
significantly	NN	O	O
lower	NN	O	O
in	NN	O	O
females	NN	O	O
(	NN	O	O
2.32	NN	O	O
+/-	NN	O	O
0.92	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
PBMC	NN	O	B-cell_type
vs	NN	O	O
4.43	NN	O	O
+/-	NN	O	O
1.38	NN	O	O
fmol/10	NN	O	O
(	NN	O	O
7	NN	O	O
)	NN	O	O
PBMC	NN	O	B-cell_type
in	NN	O	O
males	NN	O	O
;	NN	O	O
p	NN	O	O
=	NN	O	O
0.0001	NN	O	O
)	NN	O	O
.	NN	O	O

Neither	NN	O	O
Kd	NN	O	O
nor	NN	O	O
Nmax	NN	O	O
were	NN	O	O
significantly	NN	O	O
correlated	NN	O	O
with	NN	O	O
age	NN	O	O
.	NN	O	O

No	NN	O	O
difference	NN	O	O
was	NN	O	O
found	NN	O	O
between	NN	O	O
pre	NN	O	O
and	NN	O	O
postmenopausal	NN	O	O
women	NN	O	O
.	NN	O	O

Further	NN	O	O
studies	NN	O	O
are	NN	O	O
needed	NN	O	O
to	NN	O	O
elucidate	NN	O	O
if	NN	O	O
this	NN	O	O
sex	NN	O	O
difference	NN	O	O
in	NN	O	O
PBMC	NN	O	B-protein
receptors	NN	O	I-protein
for	NN	O	O
1.25	NN	O	O
Dihydroxyvitamin	NN	O	O
D3	NN	O	O
is	NN	O	O
of	NN	O	O
any	NN	O	O
pathophysiological	NN	O	O
relevance	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
Id2	NN	O	O
and	NN	O	O
Id3	NN	O	O
mRNA	NN	O	O
in	NN	O	O
human	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
.	NN	O	O

Helix-loop-helix	NN	O	B-protein
(	NN	O	I-protein
HLH	NN	O	I-protein
)	NN	O	I-protein
transcription	NN	O	I-protein
factors	NN	O	I-protein
are	NN	O	O
involved	NN	O	O
in	NN	O	O
cellular	NN	O	O
growth	NN	O	O
and	NN	O	O
differentiation	NN	O	O
.	NN	O	O

The	NN	O	O
Id	NN	O	B-protein
(	NN	O	O
inhibitor	NN	O	O
of	NN	O	O
DNA	NN	O	O
binding	NN	O	O
and	NN	O	O
differentiation	NN	O	O
)	NN	O	O
HLH	NN	O	B-protein
proteins	NN	O	I-protein
,	NN	O	O
in	NN	O	O
a	NN	O	O
dominantly	NN	O	O
negative	NN	O	O
fashion	NN	O	O
,	NN	O	O
regulate	NN	O	O
transcriptional	NN	O	O
activities	NN	O	O
of	NN	O	O
basic	NN	O	B-protein
HLH	NN	O	I-protein
proteins	NN	O	I-protein
.	NN	O	O

We	NN	O	O
examined	NN	O	O
by	NN	O	O
northern	NN	O	O
hybridization	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
Id2	NN	O	B-RNA
and	NN	O	I-RNA
Id3	NN	O	I-RNA
mRNA	NN	O	I-RNA
in	NN	O	O
human	NN	O	B-cell_line
leukemia/lymphoma	NN	O	I-cell_line
lines	NN	O	I-cell_line
and	NN	O	O
patient	NN	O	O
samples	NN	O	O
,	NN	O	O
as	NN	O	O
well	NN	O	O
as	NN	O	O
resting	NN	O	O
and	NN	O	O
activated	NN	O	B-cell_line
normal	NN	O	I-cell_line
human	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
PBL	NN	O	B-cell_type
)	NN	O	O
.	NN	O	O

The	NN	O	O
Id2	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
abundantly	NN	O	O
expressed	NN	O	O
in	NN	O	O
5/12	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
and	NN	O	O
3/4	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
Id3	NN	O	B-RNA
mRNA	NN	O	I-RNA
was	NN	O	O
detected	NN	O	O
in	NN	O	O
4/12	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
and	NN	O	O
3/4	NN	O	B-cell_line
B-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Interestingly	NN	O	O
,	NN	O	O
Id2	NN	O	B-RNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
Id3	NN	O	B-RNA
,	NN	O	O
mRNA	NN	O	O
was	NN	O	O
strongly	NN	O	O
expressed	NN	O	O
in	NN	O	O
4/5	NN	O	B-cell_line
T-cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
infected	NN	O	O
with	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
ATL-1k	NN	O	B-cell_line
,	NN	O	O
MT-2	NN	O	B-cell_line
,	NN	O	O
S-LB1	NN	O	B-cell_line
)	NN	O	O
and	NN	O	O
type	NN	O	O
II	NN	O	O
(	NN	O	O
Mo	NN	O	B-cell_line
)	NN	O	O
.	NN	O	O

Another	NN	O	O
unexpected	NN	O	O
finding	NN	O	O
was	NN	O	O
that	NN	O	O
T-cell	NN	O	O
leukemias	NN	O	O
and	NN	O	O
T-cell	NN	O	B-cell_line
lines	NN	O	I-cell_line
often	NN	O	O
expressed	NN	O	O
either	NN	O	O
Id2	NN	O	B-RNA
or	NN	O	O
Id3	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

In	NN	O	O
addition	NN	O	O
,	NN	O	O
resting	NN	O	B-cell_type
PBL	NN	O	I-cell_type
constitutively	NN	O	O
expressed	NN	O	O
prominent	NN	O	O
levels	NN	O	O
of	NN	O	O
Id2	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
but	NN	O	O
not	NN	O	O
Id3	NN	O	B-RNA
mRNA	NN	O	I-RNA
.	NN	O	O

Upon	NN	O	O
PHA	NN	O	B-protein
-stimulation	NN	O	O
,	NN	O	O
Id2	NN	O	B-protein
expression	NN	O	O
decreased	NN	O	O
and	NN	O	O
Id3	NN	O	B-protein
levels	NN	O	O
increased	NN	O	O
with	NN	O	O
biphasic	NN	O	O
kinetics	NN	O	O
.	NN	O	O

Taken	NN	O	O
together	NN	O	O
,	NN	O	O
our	NN	O	O
studies	NN	O	O
revealed	NN	O	O
three	NN	O	O
unexpected	NN	O	O
findings	NN	O	O
which	NN	O	O
require	NN	O	O
further	NN	O	O
analysis	NN	O	O
:	NN	O	O
(	NN	O	O
1	NN	O	O
)	NN	O	O
expression	NN	O	O
of	NN	O	O
Id2	NN	O	B-RNA
mRNA	NN	O	I-RNA
is	NN	O	O
often	NN	O	O
associated	NN	O	O
with	NN	O	O
lymphocytic	NN	O	O
transformation	NN	O	O
by	NN	O	O
HTLV-I	NN	O	O
or	NN	O	O
-II	NN	O	O
;	NN	O	O
(	NN	O	O
2	NN	O	O
)	NN	O	O
T-cells	NN	O	O
usually	NN	O	O
express	NN	O	O
either	NN	O	O
Id2	NN	O	B-protein
or	NN	O	O
Id3	NN	O	B-RNA
mRNA	NN	O	I-RNA
,	NN	O	O
but	NN	O	O
B-cells	NN	O	O
often	NN	O	O
express	NN	O	O
both	NN	O	O
simultaneously	NN	O	O
;	NN	O	O
(	NN	O	O
3	NN	O	O
)	NN	O	O
non-dividing	NN	O	O
,	NN	O	O
normal	NN	O	O
PBL	NN	O	B-cell_type
express	NN	O	O
high	NN	O	O
levels	NN	O	O
of	NN	O	O
Id2	NN	O	B-protein
and	NN	O	O
no	NN	O	O
Id3	NN	O	B-RNA
mRNA	NN	O	I-RNA
;	NN	O	O
and	NN	O	O
with	NN	O	O
the	NN	O	O
onset	NN	O	O
of	NN	O	O
cellular	NN	O	O
proliferation	NN	O	O
,	NN	O	O
levels	NN	O	O
of	NN	O	O
Id2	NN	O	B-RNA
mRNA	NN	O	I-RNA
decrease	NN	O	O
while	NN	O	O
levels	NN	O	O
of	NN	O	O
Id3	NN	O	B-RNA
mRNA	NN	O	I-RNA
increase	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
regulation	NN	O	O
of	NN	O	O
expression	NN	O	O
of	NN	O	O
these	NN	O	O
closely	NN	O	O
related	NN	O	O
genes	NN	O	B-DNA
is	NN	O	O
disparate	NN	O	O
.	NN	O	O

-DOCSTART-	O

Salicylates	NN	O	O
inhibit	NN	O	O
lipopolysaccharide-induced	NN	O	O
transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
tissue	NN	O	O
factor	NN	O	O
gene	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

Binding	NN	O	O
of	NN	O	O
plasma	NN	O	B-protein
Factor	NN	O	I-protein
VII/VIIa	NN	O	I-protein
to	NN	O	O
the	NN	O	O
tissue	NN	O	B-protein
factor	NN	O	I-protein
(	NN	O	I-protein
TF	NN	O	I-protein
)	NN	O	I-protein
receptor	NN	O	I-protein
initiates	NN	O	O
the	NN	O	O
coagulation	NN	O	B-protein
protease	NN	O	I-protein
cascades	NN	O	O
.	NN	O	O

TF	NN	O	O
expression	NN	O	O
by	NN	O	O
circulating	NN	O	B-cell_type
monocytes	NN	O	I-cell_type
is	NN	O	O
associated	NN	O	O
with	NN	O	O
thrombotic	NN	O	O
and	NN	O	O
inflammatory	NN	O	O
complications	NN	O	O
in	NN	O	O
a	NN	O	O
variety	NN	O	O
of	NN	O	O
diseases	NN	O	O
.	NN	O	O

Transcriptional	NN	O	O
activation	NN	O	O
of	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
TF	NN	O	I-DNA
gene	NN	O	I-DNA
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
exposed	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
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	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

Here	NN	O	O
,	NN	O	O
we	NN	O	O
report	NN	O	O
that	NN	O	O
a	NN	O	O
family	NN	O	O
of	NN	O	O
anti-inflammatory	NN	O	O
agents	NN	O	O
,	NN	O	O
known	NN	O	O
as	NN	O	O
the	NN	O	O
salicylates	NN	O	O
,	NN	O	O
inhibited	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
TF	NN	O	O
activity	NN	O	O
and	NN	O	O
TF	NN	O	B-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
and	NN	O	O
monocytic	NN	O	B-cell_line
THP-1	NN	O	I-cell_line
cells	NN	O	I-cell_line
at	NN	O	O
clinically	NN	O	O
relevant	NN	O	O
doses	NN	O	O
.	NN	O	O

Furthermore	NN	O	O
,	NN	O	O
sodium	NN	O	O
salicylate	NN	O	O
blocked	NN	O	O
the	NN	O	O
LPS-induced	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
which	NN	O	O
prevented	NN	O	O
the	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
two	NN	O	O
other	NN	O	O
nonsteroidal	NN	O	O
anti-inflammatory	NN	O	O
drugs	NN	O	O
,	NN	O	O
ibuprofen	NN	O	O
and	NN	O	O
indomethacin	NN	O	O
,	NN	O	O
did	NN	O	O
not	NN	O	O
inhibit	NN	O	O
LPS	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

These	NN	O	O
results	NN	O	O
indicated	NN	O	O
that	NN	O	O
salicylates	NN	O	O
inhibited	NN	O	O
LPS	NN	O	O
induction	NN	O	O
of	NN	O	O
TF	NN	O	B-DNA
gene	NN	O	I-DNA
transcription	NN	O	O
in	NN	O	O
monocytic	NN	O	B-cell_type
cells	NN	O	I-cell_type
by	NN	O	O
preventing	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

The	NN	O	O
clinical	NN	O	O
benefits	NN	O	O
of	NN	O	O
salicylates	NN	O	O
in	NN	O	O
the	NN	O	O
treatment	NN	O	O
of	NN	O	O
several	NN	O	O
diseases	NN	O	O
,	NN	O	O
including	NN	O	O
atherosclerosis	NN	O	O
and	NN	O	O
rheumatoid	NN	O	O
arthritis	NN	O	O
,	NN	O	O
may	NN	O	O
be	NN	O	O
related	NN	O	O
to	NN	O	O
their	NN	O	O
ability	NN	O	O
to	NN	O	O
reduce	NN	O	O
monocyte	NN	O	O
gene	NN	O	O
expression	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
signal	NN	O	O
transducer	NN	O	O
and	NN	O	O
transcription	NN	O	O
(	NN	O	O
STAT	NN	O	B-protein
)	NN	O	O
signaling	NN	O	O
pathway	NN	O	O
in	NN	O	O
a	NN	O	O
primary	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
.	NN	O	O

Critical	NN	O	O
role	NN	O	O
for	NN	O	O
IL-6	NN	O	B-protein
.	NN	O	O

The	NN	O	O
T	NN	O	O
cell	NN	O	O
activation	NN	O	O
is	NN	O	O
initiated	NN	O	O
by	NN	O	O
interaction	NN	O	O
of	NN	O	O
specific	NN	O	O
Ags	NN	O	B-protein
with	NN	O	O
TCR	NN	O	B-protein
,	NN	O	O
followed	NN	O	O
by	NN	O	O
activation	NN	O	O
of	NN	O	O
intracellular	NN	O	O
biochemical	NN	O	O
events	NN	O	O
leading	NN	O	O
to	NN	O	O
activation	NN	O	O
of	NN	O	O
several	NN	O	O
genes	NN	O	O
.	NN	O	O

The	NN	O	O
activation	NN	O	O
of	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
proteins	NN	O	I-protein
in	NN	O	O
a	NN	O	O
primary	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
activation	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
have	NN	O	O
been	NN	O	O
explored	NN	O	O
.	NN	O	O

In	NN	O	O
purified	NN	O	B-cell_type
human	NN	O	I-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
nuclear	NN	O	B-protein
STAT	NN	O	I-protein
proteins	NN	O	I-protein
were	NN	O	O
activated	NN	O	O
approximately	NN	O	O
3	NN	O	O
h	NN	O	O
after	NN	O	O
activation	NN	O	O
by	NN	O	O
cross-linked	NN	O	O
anti-CD3	NN	O	B-protein
Abs	NN	O	I-protein
.	NN	O	O

These	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
were	NN	O	O
detected	NN	O	O
by	NN	O	O
using	NN	O	O
the	NN	O	O
IFN-gamma-activated	NN	O	B-DNA
sequence	NN	O	I-DNA
(	NN	O	O
GAS	NN	O	B-DNA
)	NN	O	O
and	NN	O	O
related	NN	O	O
oligonucleotides	NN	O	O
as	NN	O	O
probes	NN	O	O
in	NN	O	O
electrophoretic	NN	O	O
mobility	NN	O	O
shift	NN	O	O
assay	NN	O	O
.	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
anti-STAT	NN	O	B-protein
Abs	NN	O	I-protein
indicated	NN	O	O
that	NN	O	O
they	NN	O	O
contained	NN	O	O
STAT-3	NN	O	B-protein
and	NN	O	O
additional	NN	O	O
proteins	NN	O	O
crossreactive	NN	O	O
with	NN	O	O
the	NN	O	O
STAT	NN	O	B-protein
family	NN	O	I-protein
.	NN	O	O

The	NN	O	O
induction	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
activity	NN	O	O
was	NN	O	O
inhibited	NN	O	O
completely	NN	O	O
by	NN	O	O
pretreatment	NN	O	O
with	NN	O	O
either	NN	O	O
cycloheximide	NN	O	O
or	NN	O	O
cyclosporin	NN	O	O
A	NN	O	O
,	NN	O	O
thus	NN	O	O
indicating	NN	O	O
that	NN	O	O
the	NN	O	O
induction	NN	O	O
was	NN	O	O
due	NN	O	O
to	NN	O	O
a	NN	O	O
secondary	NN	O	B-protein
factor	NN	O	I-protein
produced	NN	O	O
by	NN	O	O
the	NN	O	O
activated	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
.	NN	O	O

As	NN	O	O
neutralizing	NN	O	B-protein
anti-IL-6	NN	O	I-protein
Abs	NN	O	I-protein
effectively	NN	O	O
down-regulated	NN	O	O
the	NN	O	O
early	NN	O	O
induction	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
and	NN	O	O
as	NN	O	O
exogenously	NN	O	O
added	NN	O	O
IL-6	NN	O	B-protein
rapidly	NN	O	O
activated	NN	O	O
DNA	NN	O	O
binding	NN	O	O
similar	NN	O	O
to	NN	O	O
TCR	NN	O	B-protein
-mediated	NN	O	O
bindings	NN	O	O
,	NN	O	O
it	NN	O	O
can	NN	O	O
be	NN	O	O
concluded	NN	O	O
that	NN	O	O
IL-6	NN	O	B-protein
is	NN	O	O
the	NN	O	O
factor	NN	O	O
responsible	NN	O	O
for	NN	O	O
the	NN	O	O
activation	NN	O	O
of	NN	O	O
STAT	NN	O	B-protein
proteins	NN	O	I-protein
in	NN	O	O
a	NN	O	O
primary	NN	O	O
T	NN	O	O
cell	NN	O	O
response	NN	O	O
.	NN	O	O

-DOCSTART-	O

The	NN	O	O
normal	NN	O	O
cell	NN	O	O
cycle	NN	O	O
activation	NN	O	O
program	NN	O	O
is	NN	O	O
exploited	NN	O	O
during	NN	O	O
the	NN	O	O
infection	NN	O	O
of	NN	O	O
quiescent	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
by	NN	O	O
Epstein-Barr	NN	O	O
virus	NN	O	O
.	NN	O	O

B	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
in	NN	O	O
the	NN	O	O
peripheral	NN	O	O
circulation	NN	O	O
are	NN	O	O
maintained	NN	O	O
in	NN	O	O
a	NN	O	O
non-proliferative	NN	O	O
state	NN	O	O
.	NN	O	O

Antigen	NN	O	O
recognition	NN	O	O
stimulates	NN	O	O
limited	NN	O	O
proliferation	NN	O	O
,	NN	O	O
whereas	NN	O	O
infection	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
results	NN	O	O
in	NN	O	O
continual	NN	O	O
proliferation	NN	O	O
and	NN	O	O
the	NN	O	O
outgrowth	NN	O	O
of	NN	O	O
immortal	NN	O	B-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
.	NN	O	O

Because	NN	O	O
it	NN	O	O
is	NN	O	O
not	NN	O	O
clear	NN	O	O
at	NN	O	O
which	NN	O	O
point	NN	O	O
in	NN	O	O
cell	NN	O	O
cycle	NN	O	O
the	NN	O	O
peripheral	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
are	NN	O	O
arrested	NN	O	O
,	NN	O	O
we	NN	O	O
characterized	NN	O	O
the	NN	O	O
expression	NN	O	O
of	NN	O	O
several	NN	O	O
cell	NN	O	B-DNA
cycle-associated	NN	O	I-DNA
genes	NN	O	I-DNA
in	NN	O	O
quiescent	NN	O	B-cell_type
and	NN	O	I-cell_type
stimulated	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
the	NN	O	O
expression	NN	O	O
of	NN	O	O
four	NN	O	O
cell	NN	O	B-DNA
genes	NN	O	I-DNA
,	NN	O	O
cdc-2	NN	O	B-DNA
,	NN	O	O
cyclin	NN	O	B-DNA
E	NN	O	I-DNA
,	NN	O	O
CD23	NN	O	B-DNA
,	NN	O	O
and	NN	O	O
cyclin	NN	O	B-DNA
D2	NN	O	I-DNA
,	NN	O	O
are	NN	O	O
up-regulated	NN	O	O
approximately	NN	O	O
100-fold	NN	O	O
as	NN	O	O
a	NN	O	O
result	NN	O	O
of	NN	O	O
EBV-mediated	NN	O	O
immortalization	NN	O	O
.	NN	O	O

Because	NN	O	O
these	NN	O	O
genes	NN	O	O
play	NN	O	O
a	NN	O	O
positive	NN	O	O
role	NN	O	O
in	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
,	NN	O	O
we	NN	O	O
suggest	NN	O	O
that	NN	O	O
this	NN	O	O
regulatory	NN	O	O
switch	NN	O	O
contributes	NN	O	O
to	NN	O	O
controlling	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

Transient	NN	O	O
stimulation	NN	O	O
of	NN	O	O
quiescent	NN	O	B-cell_type
B	NN	O	I-cell_type
lymphocytes	NN	O	I-cell_type
with	NN	O	O
either	NN	O	O
a	NN	O	O
cocktail	NN	O	O
of	NN	O	O
anti-CD40	NN	O	B-protein
,	NN	O	O
anti-IgM	NN	O	B-protein
,	NN	O	O
and	NN	O	O
IL4	NN	O	B-protein
,	NN	O	O
or	NN	O	O
EBV	NN	O	O
results	NN	O	O
in	NN	O	O
the	NN	O	O
rapid	NN	O	O
expression	NN	O	O
of	NN	O	O
the	NN	O	O
same	NN	O	O
four	NN	O	O
genes	NN	O	O
,	NN	O	O
suggesting	NN	O	O
that	NN	O	O
,	NN	O	O
after	NN	O	O
infection	NN	O	O
,	NN	O	O
EBV	NN	O	O
exploits	NN	O	O
the	NN	O	O
normal	NN	O	O
program	NN	O	O
of	NN	O	O
B-lymphocyte	NN	O	B-cell_type
cell	NN	O	O
cycle	NN	O	O
activation	NN	O	O
.	NN	O	O

-DOCSTART-	O

Expression	NN	O	O
of	NN	O	O
the	NN	O	O
chemokine	NN	O	B-protein
receptor	NN	O	I-protein
BLR2/EBI1	NN	O	B-protein
is	NN	O	O
specifically	NN	O	O
transactivated	NN	O	O
by	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

In	NN	O	O
our	NN	O	O
attempt	NN	O	O
to	NN	O	O
identify	NN	O	O
chemokine	NN	O	B-protein
receptors	NN	O	I-protein
that	NN	O	O
are	NN	O	O
related	NN	O	O
to	NN	O	O
Burkitt	NN	O	B-protein
's	NN	O	I-protein
lymphoma	NN	O	I-protein
receptor	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
BLR1	NN	O	B-protein
)	NN	O	O
and	NN	O	O
are	NN	O	O
expressed	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
we	NN	O	O
used	NN	O	O
RT-PCR	NN	O	O
resulting	NN	O	O
in	NN	O	O
the	NN	O	O
isolation	NN	O	O
of	NN	O	O
a	NN	O	O
cDNA	NN	O	O
encoding	NN	O	O
a	NN	O	O
seven	NN	O	B-protein
transmembrane	NN	O	I-protein
receptor	NN	O	I-protein
termed	NN	O	O
BLR2	NN	O	B-protein
.	NN	O	O

The	NN	O	O
protein	NN	O	O
shows	NN	O	O
significant	NN	O	O
sequence	NN	O	O
similarities	NN	O	O
to	NN	O	O
the	NN	O	O
family	NN	O	O
of	NN	O	O
G-protein	NN	O	B-protein
coupled	NN	O	I-protein
chemokine	NN	O	I-protein
receptors	NN	O	I-protein
and	NN	O	O
turned	NN	O	O
out	NN	O	O
to	NN	O	O
be	NN	O	O
identical	NN	O	O
to	NN	O	O
the	NN	O	O
recently	NN	O	O
described	NN	O	O
receptor	NN	O	B-protein
EBI1	NN	O	I-protein
.	NN	O	O

Northern	NN	O	O
blot	NN	O	O
analysis	NN	O	O
revealed	NN	O	O
that	NN	O	O
BLR2	NN	O	B-RNA
mRNA	NN	O	I-RNA
could	NN	O	O
be	NN	O	O
highly	NN	O	O
stimulated	NN	O	O
in	NN	O	O
mitogen-	NN	O	B-cell_line
and	NN	O	I-cell_line
anti-CD3-treated	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

BLR2-specific	NN	O	B-RNA
mRNA	NN	O	I-RNA
could	NN	O	O
be	NN	O	O
detected	NN	O	O
in	NN	O	O
all	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus	NN	O	I-cell_line
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

We	NN	O	O
show	NN	O	O
that	NN	O	O
transcription	NN	O	O
of	NN	O	O
the	NN	O	O
BLR2	NN	O	B-DNA
gene	NN	O	I-DNA
could	NN	O	O
be	NN	O	O
specifically	NN	O	O
induced	NN	O	O
in	NN	O	O
Epstein-Barr	NN	O	B-cell_line
virus	NN	O	I-cell_line
negative	NN	O	I-cell_line
BL	NN	O	I-cell_line
41	NN	O	I-cell_line
cells	NN	O	I-cell_line
via	NN	O	O
estrogen-mediated	NN	O	O
activation	NN	O	O
of	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
a	NN	O	O
key	NN	O	O
regulator	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
in	NN	O	O
immortalized	NN	O	B-cell_line
B	NN	O	I-cell_line
cells	NN	O	I-cell_line
.	NN	O	O

Our	NN	O	O
data	NN	O	O
suggest	NN	O	O
an	NN	O	O
involvement	NN	O	O
of	NN	O	O
BLR2	NN	O	B-protein
in	NN	O	O
the	NN	O	O
regulation	NN	O	O
of	NN	O	O
migration	NN	O	O
in	NN	O	O
activated	NN	O	B-cell_type
lymphocytes	NN	O	I-cell_type
and	NN	O	O
in	NN	O	O
viral	NN	O	O
pathogenesis	NN	O	O
.	NN	O	O

-DOCSTART-	O

A	NN	O	O
central	NN	O	O
role	NN	O	O
for	NN	O	O
a	NN	O	O
single	NN	O	B-DNA
c-Myb	NN	O	I-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
thymic	NN	O	B-DNA
locus	NN	O	I-DNA
control	NN	O	I-DNA
region	NN	O	I-DNA
.	NN	O	O

Locus	NN	O	B-DNA
control	NN	O	I-DNA
regions	NN	O	I-DNA
(	NN	O	O
LCRs	NN	O	B-DNA
)	NN	O	O
are	NN	O	O
powerful	NN	O	O
assemblies	NN	O	O
of	NN	O	O
cis	NN	O	B-DNA
elements	NN	O	I-DNA
that	NN	O	O
organize	NN	O	O
the	NN	O	O
actions	NN	O	O
of	NN	O	O
cell-type-specific	NN	O	B-protein
trans-acting	NN	O	I-protein
factors	NN	O	I-protein
.	NN	O	O

A	NN	O	O
2.3-kb	NN	O	B-protein
LCR	NN	O	I-protein
in	NN	O	O
the	NN	O	O
human	NN	O	B-DNA
adenosine	NN	O	I-DNA
deaminase	NN	O	I-DNA
(	NN	O	I-DNA
ADA	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
first	NN	O	I-DNA
intron	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
controls	NN	O	O
expression	NN	O	O
in	NN	O	O
thymocytes	NN	O	B-cell_type
,	NN	O	O
is	NN	O	O
composed	NN	O	O
of	NN	O	O
a	NN	O	O
200-bp	NN	O	B-DNA
enhancer	NN	O	I-DNA
domain	NN	O	I-DNA
and	NN	O	O
extended	NN	O	O
flanking	NN	O	O
sequences	NN	O	O
that	NN	O	O
facilitate	NN	O	O
activation	NN	O	O
from	NN	O	O
within	NN	O	O
chromatin	NN	O	B-DNA
.	NN	O	O

Prior	NN	O	O
analyses	NN	O	O
have	NN	O	O
demonstrated	NN	O	O
that	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
contains	NN	O	O
a	NN	O	O
28-bp	NN	O	B-DNA
core	NN	O	I-DNA
region	NN	O	I-DNA
and	NN	O	O
local	NN	O	O
adjacent	NN	O	O
augmentative	NN	O	O
cis	NN	O	B-DNA
elements	NN	O	I-DNA
.	NN	O	O

We	NN	O	O
now	NN	O	O
show	NN	O	O
that	NN	O	O
the	NN	O	O
core	NN	O	O
contains	NN	O	O
a	NN	O	O
single	NN	O	O
critical	NN	O	O
c-Myb	NN	O	B-DNA
binding	NN	O	I-DNA
site	NN	O	I-DNA
.	NN	O	O

In	NN	O	O
both	NN	O	O
transiently	NN	O	B-cell_line
cotransfected	NN	O	I-cell_line
human	NN	O	I-cell_line
cells	NN	O	I-cell_line
and	NN	O	O
stable	NN	O	O
chromatin-integrated	NN	O	B-cell_line
yeast	NN	O	I-cell_line
cells	NN	O	I-cell_line
,	NN	O	O
c-Myb	NN	O	B-protein
strongly	NN	O	O
transactivated	NN	O	B-DNA
reporter	NN	O	I-DNA
constructs	NN	O	I-DNA
that	NN	O	O
contained	NN	O	O
polymerized	NN	O	B-DNA
core	NN	O	I-DNA
sequences	NN	O	I-DNA
.	NN	O	O

c-Myb	NN	O	B-protein
protein	NN	O	I-protein
was	NN	O	O
strongly	NN	O	O
evident	NN	O	O
in	NN	O	O
T	NN	O	B-cell_type
lymphoblasts	NN	O	I-cell_type
in	NN	O	O
which	NN	O	O
the	NN	O	O
enhancer	NN	O	B-DNA
was	NN	O	O
active	NN	O	O
and	NN	O	O
was	NN	O	O
localized	NN	O	O
within	NN	O	O
discrete	NN	O	O
nuclear	NN	O	O
structures	NN	O	O
.	NN	O	O

Fetal	NN	O	O
murine	NN	O	O
thymus	NN	O	O
exhibited	NN	O	O
a	NN	O	O
striking	NN	O	O
concordance	NN	O	O
of	NN	O	O
endogenous	NN	O	O
c-myb	NN	O	B-DNA
expression	NN	O	O
with	NN	O	O
that	NN	O	O
of	NN	O	O
mouse	NN	O	B-DNA
ADA	NN	O	I-DNA
and	NN	O	O
human	NN	O	B-DNA
ADA	NN	O	I-DNA
LCR	NN	O	I-DNA
-directed	NN	O	O
transgene	NN	O	O
expression	NN	O	O
.	NN	O	O

Point	NN	O	O
mutation	NN	O	O
of	NN	O	O
the	NN	O	O
c-Myb	NN	O	B-DNA
site	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
intact	NN	O	O
2.3-kb	NN	O	B-protein
LCR	NN	O	I-protein
severely	NN	O	O
attenuated	NN	O	O
enhancer	NN	O	O
activity	NN	O	O
in	NN	O	O
transfections	NN	O	O
and	NN	O	O
LCR	NN	O	B-DNA
activity	NN	O	O
in	NN	O	O
transgenic	NN	O	B-cell_line
thymocytes	NN	O	I-cell_line
.	NN	O	O

Within	NN	O	O
the	NN	O	O
context	NN	O	O
of	NN	O	O
a	NN	O	O
complex	NN	O	B-DNA
enhancer	NN	O	I-DNA
and	NN	O	O
LCR	NN	O	B-DNA
,	NN	O	O
c-Myb	NN	O	B-protein
can	NN	O	O
act	NN	O	O
as	NN	O	O
an	NN	O	O
organizer	NN	O	O
of	NN	O	O
thymocyte-specific	NN	O	B-DNA
gene	NN	O	I-DNA
expression	NN	O	O
via	NN	O	O
a	NN	O	O
single	NN	O	O
binding	NN	O	O
site	NN	O	O
.	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
human	NN	O	B-DNA
interleukin	NN	O	I-DNA
2	NN	O	I-DNA
gene	NN	O	I-DNA
promoter	NN	O	I-DNA
is	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
zinc	NN	O	B-protein
finger	NN	O	I-protein
proteins	NN	O	I-protein
Sp1	NN	O	B-protein
and	NN	O	O
EGR-1	NN	O	B-protein
.	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
(	NN	O	I-DNA
IL-2	NN	O	I-DNA
)	NN	O	I-DNA
gene	NN	O	I-DNA
after	NN	O	O
antigen	NN	O	O
recognition	NN	O	O
is	NN	O	O
a	NN	O	O
critical	NN	O	O
event	NN	O	O
for	NN	O	O
T	NN	O	O
cell	NN	O	O
proliferation	NN	O	O
and	NN	O	O
effector	NN	O	O
function	NN	O	O
.	NN	O	O

Prior	NN	O	O
studies	NN	O	O
have	NN	O	O
identified	NN	O	O
several	NN	O	O
transcription	NN	O	B-protein
factors	NN	O	I-protein
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
stimulated	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
describe	NN	O	O
a	NN	O	O
novel	NN	O	O
regulatory	NN	O	B-DNA
element	NN	O	I-DNA
within	NN	O	O
the	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
located	NN	O	O
immediately	NN	O	O
upstream	NN	O	O
of	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
(	NN	O	I-DNA
NFAT	NN	O	I-DNA
)	NN	O	I-DNA
domain	NN	O	I-DNA
.	NN	O	O

This	NN	O	O
region	NN	O	O
(	NN	O	O
termed	NN	O	O
the	NN	O	O
zinc	NN	O	B-DNA
finger	NN	O	I-DNA
protein	NN	O	I-DNA
binding	NN	O	I-DNA
region	NN	O	I-DNA
(	NN	O	O
ZIP	NN	O	B-DNA
)	NN	O	O
)	NN	O	O
serves	NN	O	O
as	NN	O	O
binding	NN	O	O
site	NN	O	O
for	NN	O	O
two	NN	O	O
differently	NN	O	O
regulated	NN	O	O
zinc	NN	O	B-protein
finger	NN	O	I-protein
proteins	NN	O	I-protein
:	NN	O	O
the	NN	O	O
constitutively	NN	O	B-protein
expressed	NN	O	I-protein
transcription	NN	O	I-protein
factor	NN	O	I-protein
Sp1	NN	O	B-protein
and	NN	O	O
the	NN	O	O
inducible	NN	O	B-protein
early	NN	O	I-protein
growth	NN	O	I-protein
response	NN	O	I-protein
protein	NN	O	I-protein
EGR-1	NN	O	B-protein
.	NN	O	O

In	NN	O	O
unstimulated	NN	O	B-cell_type
cells	NN	O	I-cell_type
which	NN	O	O
do	NN	O	O
not	NN	O	O
secrete	NN	O	O
IL-2	NN	O	B-protein
,	NN	O	O
only	NN	O	O
Sp1	NN	O	B-protein
binds	NN	O	O
to	NN	O	O
this	NN	O	O
region	NN	O	O
,	NN	O	O
while	NN	O	O
in	NN	O	O
stimulated	NN	O	B-cell_line
IL-2	NN	O	I-cell_line
secreting	NN	O	I-cell_line
cells	NN	O	I-cell_line
the	NN	O	O
inducible	NN	O	B-protein
EGR-1	NN	O	I-protein
protein	NN	O	I-protein
recognizes	NN	O	O
this	NN	O	O
element	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
the	NN	O	O
ZIP	NN	O	B-DNA
site	NN	O	I-DNA
serves	NN	O	O
as	NN	O	O
an	NN	O	O
activator	NN	O	O
for	NN	O	O
IL-2	NN	O	B-protein
gene	NN	O	O
expression	NN	O	O
,	NN	O	O
and	NN	O	O
a	NN	O	O
combination	NN	O	O
of	NN	O	O
ZIP	NN	O	B-DNA
and	NN	O	O
NFAT	NN	O	B-DNA
binding	NN	O	I-DNA
sites	NN	O	I-DNA
is	NN	O	O
required	NN	O	O
for	NN	O	O
maximal	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
results	NN	O	O
suggest	NN	O	O
a	NN	O	O
critical	NN	O	O
role	NN	O	O
of	NN	O	O
the	NN	O	O
ZIP	NN	O	B-DNA
site	NN	O	I-DNA
for	NN	O	O
IL-2	NN	O	B-DNA
promoter	NN	O	I-DNA
activity	NN	O	O
.	NN	O	O

-DOCSTART-	O

Activation	NN	O	O
of	NN	O	O
the	NN	O	O
HIV-1	NN	O	B-DNA
enhancer	NN	O	I-DNA
by	NN	O	O
the	NN	O	O
LEF-1	NN	O	B-protein
HMG	NN	O	I-protein
protein	NN	O	I-protein
on	NN	O	O
nucleosome-assembled	NN	O	B-DNA
DNA	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

Lymphoid	NN	O	B-protein
enhancer-binding	NN	O	I-protein
factor	NN	O	I-protein
1	NN	O	I-protein
(	NN	O	O
LEF-1	NN	O	B-protein
)	NN	O	O
is	NN	O	O
a	NN	O	O
regulatory	NN	O	B-protein
high	NN	O	I-protein
mobility	NN	O	I-protein
group	NN	O	I-protein
(	NN	O	I-protein
HMG	NN	O	I-protein
)	NN	O	I-protein
protein	NN	O	I-protein
that	NN	O	O
activates	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	NN	O	I-DNA
alpha	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
in	NN	O	O
a	NN	O	O
context-restricted	NN	O	O
manner	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
this	NN	O	O
paper	NN	O	O
we	NN	O	O
demonstrate	NN	O	O
that	NN	O	O
the	NN	O	O
distal	NN	O	O
region	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
(	NN	O	I-DNA
HIV-1	NN	O	I-DNA
)	NN	O	I-DNA
enhancer	NN	O	I-DNA
,	NN	O	O
which	NN	O	O
contains	NN	O	O
DNA-binding	NN	O	B-DNA
sites	NN	O	I-DNA
for	NN	O	O
LEF-1	NN	O	B-protein
and	NN	O	O
Ets-1	NN	O	O
,	NN	O	O
also	NN	O	O
provides	NN	O	O
a	NN	O	O
functional	NN	O	O
context	NN	O	O
for	NN	O	O
activation	NN	O	O
by	NN	O	O
LEF-1	NN	O	B-protein
.	NN	O	O

First	NN	O	O
,	NN	O	O
we	NN	O	O
show	NN	O	O
that	NN	O	O
mutations	NN	O	O
in	NN	O	O
the	NN	O	O
LEF-1-binding	NN	O	B-DNA
site	NN	O	I-DNA
inhibit	NN	O	O
the	NN	O	O
activity	NN	O	O
of	NN	O	O
multimerized	NN	O	B-DNA
copies	NN	O	I-DNA
of	NN	O	O
the	NN	O	O
HIV-1	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
cells	NN	O	I-cell_line
,	NN	O	O
and	NN	O	O
that	NN	O	O
LEF-1/GAL4	NN	O	B-protein
can	NN	O	O
activate	NN	O	O
a	NN	O	O
GAL4-substituted	NN	O	B-DNA
HIV-1	NN	O	I-DNA
enhancer	NN	O	I-DNA
80-	NN	O	O
to	NN	O	O
100-fold	NN	O	O
in	NN	O	O
vivo	NN	O	O
.	NN	O	O

Second	NN	O	O
,	NN	O	O
recombinant	NN	O	B-protein
LEF-1	NN	O	I-protein
is	NN	O	O
shown	NN	O	O
to	NN	O	O
activate	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
on	NN	O	O
chromatin-assembled	NN	O	B-DNA
DNA	NN	O	I-DNA
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

By	NN	O	O
using	NN	O	O
a	NN	O	O
nucleosome	NN	O	B-protein
-assembly	NN	O	O
system	NN	O	O
derived	NN	O	O
from	NN	O	O
Drosophila	NN	O	O
embryos	NN	O	O
,	NN	O	O
we	NN	O	O
find	NN	O	O
that	NN	O	O
the	NN	O	O
packaging	NN	O	O
of	NN	O	O
DNA	NN	O	O
into	NN	O	O
chromatin	NN	O	B-DNA
in	NN	O	O
vitro	NN	O	O
strongly	NN	O	O
represses	NN	O	O
HIV-1	NN	O	O
transcription	NN	O	O
and	NN	O	O
that	NN	O	O
repression	NN	O	O
can	NN	O	O
be	NN	O	O
counteracted	NN	O	O
efficiently	NN	O	O
by	NN	O	O
preincubation	NN	O	O
of	NN	O	O
the	NN	O	O
DNA	NN	O	O
with	NN	O	O
LEF-1	NN	O	B-protein
(	NN	O	O
or	NN	O	O
LEF-1	NN	O	B-protein
and	NN	O	O
Ets-1	NN	O	B-protein
)	NN	O	O
supplemented	NN	O	O
with	NN	O	O
fractions	NN	O	O
containing	NN	O	O
the	NN	O	O
promoter-binding	NN	O	B-protein
protein	NN	O	I-protein
,	NN	O	O
Sp1	NN	O	B-protein
.	NN	O	O

Addition	NN	O	O
of	NN	O	O
TFE-3	NN	O	B-protein
,	NN	O	O
which	NN	O	O
binds	NN	O	O
to	NN	O	O
an	NN	O	O
E-box	NN	O	B-DNA
motif	NN	O	I-DNA
upstream	NN	O	O
of	NN	O	O
the	NN	O	O
LEF-1	NN	O	B-DNA
and	NN	O	I-DNA
Ets-1	NN	O	I-DNA
sites	NN	O	I-DNA
,	NN	O	O
further	NN	O	O
augments	NN	O	O
transcription	NN	O	O
in	NN	O	O
this	NN	O	O
system	NN	O	O
.	NN	O	O

Individually	NN	O	O
or	NN	O	O
collectively	NN	O	O
,	NN	O	O
none	NN	O	O
of	NN	O	O
the	NN	O	O
three	NN	O	O
enhancer-binding	NN	O	B-protein
proteins	NN	O	I-protein
(	NN	O	O
LEF-1	NN	O	B-protein
,	NN	O	O
Ets-1	NN	O	B-protein
,	NN	O	O
and	NN	O	O
TFE-3	NN	O	B-protein
)	NN	O	O
could	NN	O	O
activate	NN	O	O
transcription	NN	O	O
in	NN	O	O
the	NN	O	O
absence	NN	O	O
of	NN	O	O
Sp1	NN	O	B-protein
.	NN	O	O

A	NN	O	O
truncation	NN	O	B-protein
mutant	NN	O	I-protein
of	NN	O	O
LEF-1	NN	O	B-protein
(	NN	O	O
HMG-88	NN	O	B-protein
)	NN	O	O
,	NN	O	O
which	NN	O	O
contains	NN	O	O
the	NN	O	O
HMG	NN	O	B-protein
box	NN	O	I-protein
but	NN	O	O
lacks	NN	O	O
the	NN	O	O
trans-activation	NN	O	B-protein
domain	NN	O	I-protein
,	NN	O	O
did	NN	O	O
not	NN	O	O
activate	NN	O	O
transcription	NN	O	O
from	NN	O	O
nucleosomal	NN	O	B-DNA
DNA	NN	O	I-DNA
,	NN	O	O
indicating	NN	O	O
that	NN	O	O
bending	NN	O	O
of	NN	O	O
DNA	NN	O	O
by	NN	O	O
the	NN	O	O
HMG	NN	O	B-protein
domain	NN	O	I-protein
is	NN	O	O
not	NN	O	O
sufficient	NN	O	O
to	NN	O	O
activate	NN	O	O
transcription	NN	O	O
in	NN	O	O
vitro	NN	O	O
.	NN	O	O

We	NN	O	O
conclude	NN	O	O
that	NN	O	O
transcription	NN	O	O
activation	NN	O	O
by	NN	O	O
LEF-1	NN	O	B-protein
in	NN	O	O
vitro	NN	O	O
is	NN	O	O
a	NN	O	O
chromatin	NN	O	B-DNA
-dependent	NN	O	O
process	NN	O	O
that	NN	O	O
requires	NN	O	O
a	NN	O	O
functional	NN	O	O
trans-activation	NN	O	B-protein
domain	NN	O	I-protein
in	NN	O	O
addition	NN	O	O
to	NN	O	O
the	NN	O	O
HMG	NN	O	B-protein
domain	NN	O	I-protein
.	NN	O	O

-DOCSTART-	O

HIV-1	NN	O	B-protein
envelope	NN	O	I-protein
glycoproteins	NN	O	I-protein
induce	NN	O	O
activation	NN	O	O
of	NN	O	O
activated	NN	O	O
protein-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
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
Dec	NN	O	O
1	NN	O	O
;	NN	O	O
270	NN	O	O
(	NN	O	O
48	NN	O	O
)	NN	O	O
:	NN	O	O
29038	NN	O	O
]	NN	O	O

Activation	NN	O	O
of	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
is	NN	O	O
a	NN	O	O
primary	NN	O	O
requirement	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
entry	NN	O	O
,	NN	O	O
efficient	NN	O	O
HIV	NN	O	O
replication	NN	O	O
,	NN	O	O
and	NN	O	O
progression	NN	O	O
to	NN	O	O
AIDS	NN	O	O
,	NN	O	O
Utilizing	NN	O	O
CD4	NN	O	B-cell_line
positive	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
purified	NN	O	B-cell_type
T	NN	O	I-cell_type
cells	NN	O	I-cell_type
from	NN	O	O
normal	NN	O	O
individuals	NN	O	O
,	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	NN	O	O
,	NN	O	O
gp	NN	O	B-protein
160	NN	O	I-protein
,	NN	O	O
can	NN	O	O
induce	NN	O	O
activation	NN	O	O
of	NN	O	O
transcription	NN	O	B-protein
factor	NN	O	I-protein
,	NN	O	I-protein
activated	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

The	NN	O	O
stimulatory	NN	O	O
effects	NN	O	O
of	NN	O	O
gp160	NN	O	B-protein
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
treatment	NN	O	O
of	NN	O	O
gp160	NN	O	B-protein
with	NN	O	O
soluble	NN	O	O
CD4-IgG	NN	O	B-protein
abrogates	NN	O	O
its	NN	O	O
activity	NN	O	O
,	NN	O	O
and	NN	O	O
CD4	NN	O	B-cell_line
negative	NN	O	I-cell_line
T	NN	O	I-cell_line
cell	NN	O	I-cell_line
lines	NN	O	I-cell_line
fail	NN	O	O
to	NN	O	O
be	NN	O	O
stimulated	NN	O	O
with	NN	O	O
gp160	NN	O	B-protein
.	NN	O	O

Immunoprecipitation	NN	O	O
of	NN	O	O
the	NN	O	O
gp	NN	O	B-protein
160	NN	O	I-protein
-induced	NN	O	O
nuclear	NN	O	O
extracts	NN	O	O
with	NN	O	O
polyclonal	NN	O	B-protein
antibodies	NN	O	I-protein
to	NN	O	O
Fos	NN	O	B-protein
and	NN	O	O
Jun	NN	O	B-protein
proteins	NN	O	I-protein
indicates	NN	O	O
that	NN	O	O
AP-1	NN	O	B-protein
complex	NN	O	I-protein
is	NN	O	O
comprised	NN	O	O
of	NN	O	O
members	NN	O	O
of	NN	O	O
these	NN	O	O
family	NN	O	O
of	NN	O	O
proteins	NN	O	O
.	NN	O	O

The	NN	O	O
gp160-induced	NN	O	B-protein
AP-1	NN	O	I-protein
complex	NN	O	I-protein
is	NN	O	O
dependent	NN	O	O
upon	NN	O	O
protein	NN	O	O
tyrosine	NN	O	O
phosphorylation	NN	O	O
and	NN	O	O
is	NN	O	O
protein	NN	O	O
synthesis-independent	NN	O	O
.	NN	O	O

This	NN	O	O
stimulation	NN	O	O
can	NN	O	O
also	NN	O	O
be	NN	O	O
abolished	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
but	NN	O	O
it	NN	O	O
is	NN	O	O
unaffected	NN	O	O
by	NN	O	O
calcium	NN	O	O
channel	NN	O	O
blocker	NN	O	O
or	NN	O	O
cyclosporine	NN	O	O
A	NN	O	O
.	NN	O	O

This	NN	O	O
gp160	NN	O	B-protein
treatment	NN	O	O
adversely	NN	O	O
affects	NN	O	O
the	NN	O	O
functional	NN	O	O
capabilities	NN	O	O
of	NN	O	O
T	NN	O	B-cell_type
cells	NN	O	I-cell_type
:	NN	O	O
pre-treatment	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
with	NN	O	O
gp160	NN	O	B-protein
for	NN	O	O
4	NN	O	O
h	NN	O	O
at	NN	O	O
37	NN	O	O
degrees	NN	O	O
C	NN	O	O
inhibited	NN	O	O
anti-CD3	NN	O	B-protein
-induced	NN	O	O
interleukin-2	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

Effects	NN	O	O
similar	NN	O	O
to	NN	O	O
gp160	NN	O	B-protein
were	NN	O	O
seen	NN	O	O
with	NN	O	O
anti-CD4	NN	O	B-protein
mAb	NN	O	I-protein
.	NN	O	O

The	NN	O	O
aberrant	NN	O	O
activation	NN	O	O
of	NN	O	O
AP-1	NN	O	B-protein
by	NN	O	O
gp160	NN	O	B-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
could	NN	O	O
result	NN	O	O
in	NN	O	O
up-regulation	NN	O	O
of	NN	O	O
cytokines	NN	O	B-protein
containing	NN	O	O
AP-1	NN	O	B-DNA
sites	NN	O	I-DNA
,	NN	O	O
e.g	NN	O	O
.	NN	O	O
interleukin-3	NN	O	B-protein
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
,	NN	O	O
and	NN	O	O
concurrently	NN	O	O
lead	NN	O	O
to	NN	O	O
T	NN	O	B-cell_type
cell	NN	O	I-cell_type
unresponsiveness	NN	O	O
by	NN	O	O
inhibiting	NN	O	O
interleukin-2	NN	O	B-protein
secretion	NN	O	O
.	NN	O	O

